Agreement of Parent and Child Reports of Trauma Exposure and Symptoms in the Peritraumatic Period.
Exposure to violence and potentially traumatic events (PTE) is a common experience among children and youth. The assessment of necessary intervention relies upon parental acknowledgement of exposure and recognition of their child's distress. Early interventions and treatment are most effective when parents are aware of the nature of the traumatic exposure, understand their child's symptomatic response, and are intimately involved in the treatment process. The present study investigated concordance between parents and exposed children on child trauma history, the subjective report of the impact of the traumas experienced, and presence of PTSD symptoms. Agreement between parent and child report of traumas experienced was non-significant for serious accidents, separation from significant others, and physical assaults. Non-significant agreement was also found for avoidance and hyperarousal symptoms of PTSD. Correlations were not significant between parent and child report of the impact of traumas both at the time of the incident and at the time of the interview. Recommendations are suggested for helping parents improve their capacity to understand the potential impact of exposure on the child's psychological functioning.


Neonatal opiate withdrawal alters the reactivity of adult rats to the hot-plate.
Prenatal exposure of rats to 0.2 mg LAAM/kg/day but not to 0.05 mg LAAM/kg/day resulted in faster hot-plate escape latencies in 6 mo old offspring. No differences in tail-flick latencies were observed at 7 mo of age in offspring exposed to either dose of LAAM prenatally. Subsequent testing of littermates at 16 mo of age revealed that the greater sensitivity to the hot-plate observed in rats prenatally exposed to LAAM is apparently a result of neonatal withdrawal rather than a primary consequence of the drug. The data are discussed in relation to possible effects of drug or withdrawal on central nervous system development.


High dietary oleic acid in olive oil-supplemented diet enhanced omega-3 fatty acid in blood plasma of rats.
This study was conducted to investigate the effect of dietary oleic acid in olive oil-supplemented diets on the blood lipid profile and fatty acid composition in blood plasma and adipose tissue of rats. A total of 60 Sprague Dawley rats with mean body weight of 249 g ± 3.04 g were equally divided into three diet groups: control (CON) contained 10% coconut oil, olive50 contained 5% coconut oil and 5% olive oil, and olive100 contained 10% olive oil. Oleic acid (OA) level was highest in olive100 followed by the olive50 and control. The final body weight (BW) of the rats was significantly affected by the intake of OA, in which rats fed olive100 had the lowest final BW, which signified that OA could be associated with weight loss. Olive oil intake significantly increased levels of the high-density lipoprotein cholesterol (HDL-C) and exhibited a potential attenuation effect on the glutamic-oxaloacetic transaminase and the glutamic-pyruvic transaminase, and a potential role in the reduction of triglycerides in the bloodstream of the animals. In terms of fatty acid composition, significantly high OA was observed in the blood plasma and adipose tissues of rats fed olive100. Omega-3 polyunsaturated fatty acids (PUFAs), such as linolenic (C18:3 n-3), eicosapentaenoic (C20:5 n-3), and docosahexaenoic (C22:6 n-3), and n-6 PUFA arachidonic (C20:4 n-6) were also significantly increased in the blood plasma of rats fed olive100. These findings suggest that the intake of dietary high OA may enhance the omega-3 fatty acid levels in the blood plasma of rats and may have a positive effect in reducing risks to cardiovascular disease, as evidenced by weight loss, increased HDL-C levels, and decreased TG levels in the blood plasma of experimental animals.


HoxD transcription factors define monosynaptic sensory-motor specificity in the developing spinal cord.
The specificity of monosynaptic connections between proprioceptive sensory neurons and their recipient spinal motor neurons depends on multiple factors, including motor neuron positioning and dendrite morphology, axon projection patterns of proprioceptive sensory neurons in the spinal cord, and the ligand-receptor molecules involved in cell-to-cell recognition. However, with few exceptions, the transcription factors engaged in this process are poorly characterized. Here, we show that members of the HoxD family of transcription factors play a crucial role in the specificity of monosynaptic sensory-motor connections. Mice lacking Hoxd9, Hoxd10 and Hoxd11 exhibit defects in locomotion but have no obvious defects in motor neuron positioning or dendrite morphology through the medio-lateral and rostro-caudal axes. However, we found that quadriceps motor neurons in these mice show aberrant axon development and receive inappropriate inputs from proprioceptive sensory axons innervating the obturator muscle. These genetic studies demonstrate that the HoxD transcription factors play an integral role in the synaptic specificity of monosynaptic sensory-motor connections in the developing spinal cord.


Glucuronides from metabolites to medicines: a survey of the in vivo generation, chemical synthesis and properties of glucuronides.
Covering: 1998 to 2011. Previous review: Nat. Prod. Rep., 1998, 15, 173-186. The fourteen years that have passed since the previous review on this topic have seen a significant increase of interest in many aspects of glucuronide chemistry and biology. Glucuronides are the most important class of phase 2 xenobiotic metabolites and typically act in a detoxifying role. While this is generally true for O-alkyl and O-aryl glucuronides, a number of glucuronides are known to be pharmacologically active per se. Additionally the use of glucuronide prodrugs, notably to ameliorate the cytotoxicity of anticancer agents, has markedly increased. Whereas the previous review covered only the synthesis of O-glucuronides, we now include N-, S- and C-glucuronides also and discuss both synthetic and biological aspects. Synthetic methods for all classes of glucuronides are reviewed and updated, together with advances in the enzymatic synthesis of glucuronides and methods for their detection. Finally we discuss the biological reactivity of glucuronides where known, including the important morphine-6-glucuronide. A lively debate has continued for several years on whether O-acyl glucuronide metabolites of carboxylic acids are toxic, affecting both the safety assessment of well-used drugs and new drug development programmes. We summarise the current understanding, together with other known examples of interaction between glucuronides and macromolecules.


Infant Homicides Within the Context of Safe Haven Laws - United States, 2008-2017.
Homicide is the 13th leading cause of death among infants (i.e., children aged <1 year) in the United States (1). Infant homicides occurring within the first 24 hours of life (i.e., neonaticide) are primarily perpetrated by the mother, who might be of young age, unmarried, have lower educational attainment, and is most likely associated with concealment of an unintended pregnancy and nonhospital birthing (2). After the first day of life, infant homicides might be associated with other factors (e.g., child abuse and neglect or caregiver frustration) (2). A 2002 study of the age variation in homicide risk in U.S. infants during 1989-1998 found that the overall infant homicide rate was 8.3 per 100,000 person-years, and on the first day of life was 222.2 per 100,000 person-years, a homicide rate at least 10 times greater than that for any other time of life (3). Because of this period of heightened risk, by 2008 all 50 states* and Puerto Rico had enacted Safe Haven Laws. These laws allow a parent† to legally surrender an infant who might otherwise be abandoned or endangered (4). CDC analyzed infant homicides in the United States during 2008-2017 to determine whether rates changed after nationwide implementation of Safe Haven Laws, and to examine the association between infant homicide rates and state-specific Safe Haven age limits. During 2008-2017, the overall infant homicide rate was 7.2 per 100,000 person-years, and on the first day of life was 74.0 per 100,000 person-years, representing a 66.7% decrease from 1989-1998. However, the homicide rate on first day of life was still 5.4 times higher than that for any other time in life. No obvious association was found between infant homicide rates and Safe Haven age limits. States are encouraged to evaluate the effectiveness of their Safe Haven Laws and other prevention strategies to ensure they are achieving the intended benefits of preventing infant homicides. Programs and policies that strengthen economic supports, provide affordable childcare, and enhance and improve skills for young parents might contribute to the prevention of infant homicides.


A Novel, Selective c-Abl Inhibitor, Compound 5, Prevents Neurodegeneration in Parkinson's Disease.
Parkinson's disease (PD) is a progressive neurodegenerative disorder that affects movement. The nonreceptor tyrosine kinase c-Abl has shown a potential role in the progression of PD. As such, c-Abl inhibition is a promising candidate for neuroprotection in PD and α-synucleinopathies. Compound 5 is a newly synthesized blood-brain barrier penetrant c-Abl inhibitor with higher efficacy than existing inhibitors. The objective of the current study was to demonstrate the neuroprotective effects of compound 5 on the α-synuclein preformed fibril (α-syn PFF) mouse model of PD. Compound 5 significantly reduced neurotoxicity, activation of c-Abl, and Lewy body pathology caused by α-syn PFF in cortical neurons. Additionally, compound 5 markedly ameliorated the loss of dopaminergic neurons, c-Abl activation, Lewy body pathology, neuroinflammatory responses, and behavioral deficits induced by α-syn PFF injection in vivo. Taken together, these results suggest that compound 5 could be a pharmaceutical agent to prevent the progression of PD and α-synucleinopathies.


Chromosomal microarray and exome sequencing in unexplained early infantile epileptic encephalopathies in a highly consanguineous population.
To identify genetic causes for early infantile epileptic encephalopathies (EIEE) in Turkish children with mostly consanguineous parents. In a selected EIEE group (N = 59) based on results of nongenetic and initial genetic testing with unexplained etiology, 49 patients underwent array-based comparative genomic hybridization (aCGH) and 49 patients underwent whole exome sequencing (WES) including 39 with negative aCGH results and 10 with WES-only. Diagnostic yield of aCGH and WES for pathogenic or likely pathogenic variants was 14.3% and 38.8%, respectively. Including de novo variants of uncertain significance linked to compatible phenotypes, increased the diagnostic yield of WES to 61.2%. Out of 38 positive variants, 18 (47.4%) were novel and 16 (42.1%) were de novo. Twenty-one (56.8%) patients had recessive variants inherited from mostly consanguineous healthy parents (85.7%). Fourteen (37.8%) of patients with diagnostic results had positive variants in established EIEE genes. Seizures started during neonatal period in 32.4% patients. Posture or movement disorders were comorbid with EIEE in 40.5% of diagnosed patients. We identified treatable metabolic disorders in 8.1% of patients and pathogenic variants in genes which support using targeted medicine in 19% of patients. Detailed electro-clinical phenotyping led to expansion of some of the known phenotypes with non-neurological and neurological findings in addition to seizures, as well as suggestion of candidate genes (SEC24B, SLC16A2 and PRICKLE2) and a copy number variant (microduplication of Xp21.1p11.4). The high ratio of recessive inheritance could be important for family counseling.


Physicians and hospitals: the times demand unity.
If the voluntary health care system is to continue effectively, then physicians, hospital administrators, and trustees must cooperate in working toward common goals. Ways to enhance communication between physicians and hospitals, including more complete participation in local health systems agencies and better understanding of the mechanism of Professional Standards Review Organizations, were discussed at a conference on physician-hospital relations.


Screening potential treatments for mpox from Traditional Chinese Medicine by using a data-driven approach.
Mpox (MPX) has escalated into a public health emergency of international concern, necessitating urgent prophylactic and therapeutic measures. The primary goal of this investigation was to systematically extract Wan Quan's expertise in treating smallpox, as documented in Exclusive Methods for Treating Pox (Dou Zhen Xin Fa in Chinese), with the aim of identifying potential prescriptions, herbs, and components for alternative MPX therapies or drugs. This research utilized data mining to identify high-frequency Chinese Medicines (CMs), high-frequency CM-pairs, and CM compatibility rules. Network pharmacology, molecular docking, and molecular dynamic simulation were employed to reveal the potential molecular mechanisms of the core CM-pair. 119 prescriptions were extracted from Exclusive Methods for Treating Pox. We identified 25 high-frequency CMs and 23 high-frequency CM pairs among these prescriptions. Combined association rule mining analysis, Gancao (Glycyrrhiza uralensis Fisch.), Renshen (Panax ginseng C. A. Mey.), Danggui (Angelica sinensis (Oliv.) Diels), Shengma (Cimicifuga foetida L.), and Zicao (Lithospermum erythrorhizon Siebold & Zucc.) were selected as the core CM-pair for further investigation. Network pharmacology analysis yielded 131 active components and 348 candidate targets for the core CM-pair. Quercetin and celabenzine were chosen as ligands for molecular docking. GO and KEGG enrichment analyses revealed that the core CM-pair could interact with targets involved in immune, inflammatory, and infectious diseases. Moreover, key mpox virus targets, F8-A22-E4 DNA polymerase holoenzyme and profilin-like protein A42R, were docked well with the selected core components. And molecular dynamic simulation indicated that the component (quercetin) could stably bind to the target (profilin-like protein A42R). Our findings identified potential prescriptions, herbs, and components that can offer potential therapies or drugs for addressing the MPX epidemic.


Monoclonal antibody-induced ErbB3 receptor internalization and degradation inhibits growth and migration of human melanoma cells.
Members of the ErbB receptor family are targets of a growing numbers of small molecules and monoclonal antibodies inhibitors currently under development for the treatment of cancer. Although historical efforts have been directed against ErbB1 (EGFR) and ErbB2 (HER2/neu), emerging evidences have pointed to ErbB3 as a key node in the activation of proliferation/survival pathways from the ErbB receptor family and have fueled enthusiasm toward the clinical development of anti-ErbB3 agents. In this study, we have evaluated the potential therapeutic efficacy of a set of three recently generated anti-human ErbB3 monoclonals, A2, A3 and A4, in human primary melanoma cells. We show that in melanoma cells expressing ErbB1, ErbB3 and ErbB4 but not ErbB2 receptor ligands activate the PI3K/AKT pathway, and this leads to increased cell proliferation and migration. While antibodies A3 and A4 are able to potently inhibit ligand-induced signaling, proliferation and migration, antibody A2 is unable to exert this effect. In attempt to understand the mechanism of action and the basis of this different behavior, we demonstrate, through a series of combined approaches, that antibody efficacy strongly correlates with antibody-induced receptor internalization, degradation and inhibition of receptor recycling to the cell surface. Finally, fine epitope mapping studies through a peptide array show that inhibiting vs. non-inhibiting antibodies have a dramatically different mode of binding to the to the receptor extracellular domain. Our study confirms the key role of ErbB3 and points to exploitation of novel combination therapies for treatment of malignant melanoma.


Contrasting cumulative risk and multiple individual risk models of the relationship between Adverse Childhood Experiences (ACEs) and adult health outcomes.
A very large body of research documents relationships between self-reported Adverse Childhood Experiences (srACEs) and adult health outcomes. Despite multiple assessment tools that use the same or similar questions, there is a great deal of inconsistency in the operationalization of self-reported childhood adversity for use as a predictor variable. Alternative conceptual models are rarely used and very limited evidence directly contrasts conceptual models to each other. Also, while a cumulative numeric 'ACE Score' is normative, there are differences in the way it is calculated and used in statistical models. We investigated differences in model fit and performance between the cumulative ACE Score and a 'multiple individual risk' (MIR) model that enters individual ACE events together into prediction models. We also investigated differences that arise from the use of different strategies for coding and calculating the ACE Score. We merged the 2011-2012 BRFSS data (N = 56,640) and analyzed 3 outcomes. We compared descriptive model fit metrics and used Vuong's test for model selection to arrive at best fit models using the cumulative ACE Score (as both a continuous or categorical variable) and the MIR model, and then statistically compared the best fit models to each other. The multiple individual risk model was a better fit than the categorical ACE Score for the 'lifetime history of depression' outcome. For the outcomes of obesity and cardiac disease, the cumulative risk and multiple individual risks models were of comparable fit, but yield different and complementary inferences. Additional information-rich inferences about ACE-health relationships can be obtained from including a multiple individual risk modeling strategy. Results suggest that investigators working with large srACEs data sources could empirically derive the number of items, as well as the exposure coding strategy, that are a best fit for the outcome under study. A multiple individual risk model could also be considered in addition to the cumulative risk model, potentially in place of estimation of unadjusted ACE-outcome relationships.


Microbiological Study of Tofu.
The microbiological qualities of tofu juice and cake were studied. Seven brands of tofu from four grocery stores were tested, at day 1 and after 30 d of storage in a refrigerator. The microbial load at day 1 was different from brand to brand, but cell counts in juice and cake of the same brand were correlated. The number of cells observed at day 30 was different from brand to brand but was not related to the initial cell count. The pH had a great effect on the type of contaminating microorganisms present. All brands spoiled after 30 d of storage at 7°C; 112 isolates from both the fresh juice and cake at day 1 and at day 30 were obtained. The most common gram-positive organisms isolated were Streptococcus sp., Pediococcus sp., and Lactobacillus sp., and the most common gram-negative bacteria were Pseudomonas putida , P. aeruginosa , Enterobacter agglomerans , and E. cloacae .


Constitutional aneuploidy and cancer predisposition.
Constitutional aneuploidies are rare syndromes associated with multiple developmental abnormalities and the alterations in the risk for specific cancers. Acquired somatic chromosomal aneuploidies are the most common genetic aberrations in sporadic cancers. Thus studies of these rare constitutional aneuploidy syndromes are important not only for patient counseling and clinical management, but also for deciphering the mechanisms by which chromosomal aneuploidy affect cancer initiation and progression. Here we review the major constitutional aneuploidy syndromes and suggest some general mechanisms for the associated cancer predisposition.


Computing human to human Avian influenza ℜ₀ via transmission chains and parameter estimation.
The transmission of avian influenza between humans is extremely rare, and it mostly affects individuals who are in contact with infected family member. Although this scenario is uncommon, there have been multiple outbreaks that occur in small infection clusters in Asia with relatively lowtransmissibility, and thus are too weak to cause an epidemic. Still, subcritical transmission from stut-tering chain data is vital for determining whether avian influenza is close to the threshold of ℜ₀ > 1.In this article, we will explore two methods of estimating ℜ₀ using transmission chains and parameterestimation through data fitting. We found that ℜ₀ = 0.2205 when calculating the ℜ₀ using the maxi-mum likelihood method. When we computed the reproduction number for human to human transmis-sion through differential equations and fitted the model to data from the cumulative cases, cumulativedeaths, and cumulative secondary cases, we estimated ℜ₀ = 0.1768. To avoid violating the assumptionof the least square method, we fitted the model to incidence data to obtain ℜ₀ = 0.1520. We tested thestructural and practical identifiability of the model, and concluded that the model is identifiable undercertain assumptions. We further use two more methods to estimate ℜ₀ : by the ℜ₀ definition whichgives an overestimate of 0.28 and by Ferguson approach which yields ℜ₀ = 0.1586. We conclude that ℜ₀ for human to human transmission was about 0.2.


Proteogenomics for environmental microbiology.
Proteogenomics sensu stricto refers to the use of proteomic data to refine the annotation of genomes from model organisms. Because of the limitations of automatic annotation pipelines, a relatively high number of errors occur during the structural annotation of genes coding for proteins. Whether putative orphan sequences or short genes encoding low-molecular-weight proteins really exist is still frequently a mystery. Whether start codons are well defined is also an open debate. These problems are exacerbated for genomes of microorganisms belonging to poorly documented genera, as related sequences are not always available for homology-guided annotation. The functional annotation of a significant proportion of genes is also another well-known issue when annotating environmental microorganisms. High-throughput shotgun proteomics has recently greatly evolved, allowing the exploration of the proteome from any microorganism at an unprecedented depth. The structural and functional annotation process may be usefully complemented with experimental data. Indeed, proteogenomic mapping has been successfully performed for a wide variety of organisms. Specific approaches devoted to systematically establishing the N-termini of a large set of proteins are being developed. N-terminomics is giving rise to datasets of experimentally proven translational start codons as well as validated peptide signals for secreted proteins. By extension, combining genomic and proteomic data is becoming routine in many research projects. The proteomic analysis of organisms with unfinished genome sequences, the so-called composite proteomics, and the search for microbial biomarkers by bottom-up and top-down combined approaches are some examples of proteogenomic-flavored studies. They illustrate the advent of a new era of environmental microbiology where proteomics and genomics are intimately integrated to answer key biological questions.


The role of RhoA and cytoskeleton in myofibroblast transformation in hyperoxic lung fibrosis.
Myofibroblast transformation is a key process in the pathogenesis of lung fibrosis. We have previously reported that hyperoxia induces RhoA activation in HFL-1 lung fibroblasts and RhoA mediates collagen synthesis in hyperoxic lung fibrosis. In this study, we investigated the role of RhoA and actin cytoskeleton in hyperoxia-induced myofibroblast transformation. Exposure of HFL-1 lung fibroblasts to hyperoxia stimulated actin filament formation, shift of G-actin to F-actin, nuclear colocalization of myocardin-related transcription factor-A (MRTF-A), recruitment of MRTF-A to the α-smooth muscle actin (α-SMA) gene promoter, myofibroblast transformation, and collagen-I synthesis. Inhibition of RhoA by C3 transferase CT-04 or dominant-negative RhoA mutant T19N, and inhibition of ROCK by Y27632, prevented myofibroblast transformation and collagen-I synthesis. Moreover, inhibition of RhoA by CT-04 prevented hyperoxia-induced actin filament formation, shift of G-actin to F-actin, and nuclear colocalization of MRTF-A. In addition, disrupting actin filaments with cytochalasin D or scavenging reactive oxygen species (ROS) with tiron attenuated actin filament formation, nuclear colocalization of MRTF-A, myofibroblast transformation, and collagen-I synthesis. Furthermore, overexpression of constitutively active RhoA mutant Q63L or stabilization of actin filaments recapitulated the effects of hyperoxia on the actin cytoskeleton and nuclear colocalization of MRTF-A, myofibroblast transformation, and collagen-I synthesis. Interestingly, knocking down MRTF-A prevented hyperoxia-induced increase in the recruitment of MRTF-A to the serum response factor transcriptional complex on the α-SMA gene promoter, myofibroblast transformation, and collagen-I synthesis. Finally, Y27632 and tiron attenuated hyperoxia-induced increases in α-SMA and collagen-I in mouse lungs. Together, these results indicate that the actin cytoskeletal reorganization due to the ROS/RhoA-ROCK pathway mediates myofibroblast transformation and collagen synthesis in lung fibrosis of oxygen toxicity. MRTF-A contributes to the regulatory effect of the actin cytoskeleton on myofibroblast transformation during hyperoxia.


Optimizing financial effects of HIE: a multi-party linear programming approach.
To describe an analytical framework for quantifying the societal savings and financial consequences of a health information exchange (HIE), and to demonstrate its use in designing pricing policies for sustainable HIEs. We developed a linear programming model to (1) quantify the financial worth of HIE information to each of its participating institutions and (2) evaluate three HIE pricing policies: fixed-rate annual, charge per visit, and charge per look-up. We considered three desired outcomes of HIE-related emergency care (modeled as parameters): preventing unrequired hospitalizations, reducing duplicate tests, and avoiding emergency department (ED) visits. We applied this framework to 4639 ED encounters over a 12-month period in three large EDs in Milwaukee, Wisconsin, using Medicare/Medicaid claims data, public reports of hospital admissions, published payer mix data, and use data from a not-for-profit regional HIE. For this HIE, data accesses produced net financial gains for all providers and payers. Gains, due to HIE, were more significant for providers with more health maintenance organizations patients. Reducing unrequired hospitalizations and avoiding repeat ED visits were responsible for more than 70% of the savings. The results showed that fixed annual subscriptions can sustain this HIE, while ensuring financial gains to all participants. Sensitivity analysis revealed that the results were robust to uncertainties in modeling parameters. Our specific HIE pricing recommendations depend on the unique characteristics of this study population. However, our main contribution is the modeling approach, which is broadly applicable to other populations.


[The formation of new characteristics in life cycle of the marsh frog (Rana ridibunda) in thermal pond conditions].
Using mark and recapture approach, the long-term population dynamics in the marsh frog (Rana ridibunda) was studied. Group-marking of metamorphs was conducted in a small thermal pond serving as a sedimentation basin for discharged waters from Nizhny Tagil metallurgic works. Depending on the time of metamorphosis, three groups of individuals could be singled out, namely: early ones (group I), middle ones (group II), and late ones that overwinter as tadpoles and complete metamorphosis in May of the next year (group III). Upon metamorphosis completion, individuals of group I were found to be significantly larger than those of group II, and individuals of both these groups to be significantly smaller than those of group III. After first wintering, immature individuals from group I were significantly larger than either individuals from group II or metamorphs from group III, though a growth rate of the latter was significantly higher than in groups I and II. These discrepancies were observed both between immature and adult individuals. Over the period from metamorphosis completion to the first wintering ending, survivorship in group I was significantly higher and did not differ between groups II and III. In adult frogs, maximum survivorship was registered in group III and minimum one in group II; the detected differences recurred in each age class till the fourth wintering. However, in age classes that overwintered 4 and 5 times, maximum survivorship was observed in group II, which can be treated as a compensation for rather low survivorship of this group at younger ages. All the events of tadpoles of this species overwintering (except in other thermal water bodies) that are described in literature, correspond to rare deviations from normal ontogenesis. Therefore, the revealed formation of a numerous group of overwintering tadpoles in successive generations should be considered as a new adaptation which sense is a decrease of competition between tadpole groups when using the highly productive resources of the thermal pond practically year-round. The advantage in body size and growth rate of not only tadpoles but also of metamorphs, immature and adult individuals of group III indicates that after metamorphosis the strategy of this group still remains successful. The reason for that is unusually large body size of metamorphs which provides higher postmetamorphic survivorship and greater female fecundity.


Role of muscarinic M1 receptors in inhibitory avoidance and contextual fear conditioning.
The objective of the present study was to observe the effects of pre-training or post-training administration of dicyclomine, a M1 muscarinic antagonist, on inhibitory avoidance (IA) and contextual fear conditioning (CFC) and to investigate if the effects observed with the pre-training administration of dicyclomine are state-dependent. For each behavioral procedure (IA and CFC) groups of Wistar male rats were treated with saline or dicyclomine either 30 min before training (pre-training), immediately after training or 30 min before training/30 min before test (pre-training/pre-test). The animals were tested 24 h after training. The acquisition of IA and CFC was impaired by pre-training administration of dicyclomine. The consolidation of both tasks was not affected by dicyclomine given immediately after training. Pre-training/pre-test administration of dicyclomine impaired both tasks, an effect similar to that observed in the group which only received pre-training administration. Pre-test treatment induced dissociation between both tasks, impairing CFC retrieval, without interfering with the animals avoidance response. These results show that the dicyclomine did not affect IA and CFC consolidation, suggesting specific involvement of M1 muscarinic receptor only in acquisition these tasks, and these effects was not state-dependent. However, it is possible that the retrieval of these tasks may be mediated, at least in part, by different neurochemical mechanisms and may be dissociated by dicyclomine.


[Infection as a cause of multiple organ failure. Definition, pathophysiology and diagnostic parameters].
Several clinical observations support the hypothesis that bacterial sepsis is the main aetiological factor in at least half of the patients developing multiple organ failure. Sepsis is defined as the pathophysiological alterations and life-threatening clinical consequences of the action of microorganisms or their products invading the blood stream from a focus of infection. The clinical course of sepsis is highlighted by initial multiorgan insufficiency progressing to severe multiple system organ failure. Pathogenetic bacteria and bacterial toxins arise from the septic focus, overcome the defence mechanisms of the body, continuously invade the blood stream, activate the biological cascade systems and initiate release of mediators from blood and tissue cells. Endotoxin and activated mediators cause endothelial and organ cell dysfunction and cell damage by at least three mechanisms: maldistribution of blood flow; cytotoxia; direct inhibition of oxygen-utilising cell enzymes. The septic disease begins with unspecific signs caused by invasion of bacteria, followed by alterations of the circulatory system, the blood clotting system, the metabolism, initiating vital organ dysfunctions and finally acute respiratory, renal, gastrointestinal, hepatic failure and septic encephalopathy. The clinical diagnosis of sepsis is based on the finding of an obvious septic focus with the presence of at least four of the following 5 criteria: (I) fever above 38.8 degrees C or hypothermia below 35.5 degrees C; (II) tachypnoea (greater than 24/min) or hypocapnia (PaCO2 less than 32 mmHg); (III) tachycardia (greater than 100 Bpm), (IV) leucocytosis (greater than or equal to 15.000/mm3) or leucopenia (greater than 5.000/mm3); (V) presence of at least one indicator for inadequate organ perfusion like mental alterations, hypoxaemia (PaCO2 less than 75 mmHg while breathing room air), hyperlactataemia (greater than 1,6 mmol/l), diuresis below 30 ml/h, drop in systolic blood pressure below 100 mmHg. A positive blood culture or a positive limulus test are frequent but are not considered to be obligatory for the diagnosis of sepsis.


[Effect of x-irradiation on the generation of O2- in rat liver microsomes].
A study was made of generation of O2- in NADPH-dependent chain of oxidation of liver microsomes in irradiated rats (7 and 10 Gy). The rate of O2- generation sharply increased at early times after irradiation. The data obtained prompt an assumption that the increase in the rate of O2- generation is perhaps connected with the changes in functioning of both NADPH-cytochrome P-450-reductase and cytochrome P-450.


Training nurses for interdisciplinary communication with families in the intensive care unit: an intervention.
Critical care nurse communication training has largely been limited to didactic materials, interactive training for nurse supervisors, or brief participatory learning programs within the context of comprehensive end-of-life care educational seminars. Preliminary evidence suggests that an interactive approach can also be effective in communication skills training for intensive care unit (ICU) nurses. We implemented a 1-day educational intervention in five acute care hospitals within Veterans Integrated Service Network (VISN) 3 (New York-New Jersey region) of the Department of Veterans Affairs and focused solely on communication skills and targeted specifically to nurses providing bedside care for critically ill patients. A "learner centered" approach to skills training that has several integral components was employed. AMONG THESE ARE: a cognitive, evidence-based foundation upon which to build new skills; a method such as role-play that allows participants to practice newly learned skills; and an affective component, during which trainees can freely discuss their impressions of the exercise or explore difficulties that may have been encountered. Before and after the program we conducted a detailed assessment of participants' self-rated communication skills and of the techniques and materials we used. Post-program responses documented significant improvement in self-evaluated skills for each of the core tasks we assessed. Evidence suggests that communication with patients and families in the ICU can be most effectively approached in an interdisciplinary way. For nurses to fully realize their potential for optimal communication as members of the multidisciplinary team, they must be equipped with the necessary skills. We believe this new program helps to expand the range of approaches for training nurses in essential communication skills.


Plasma β-Endorphin Concentration and Antipsychotic Treatment Outcome in Schizophrenia: 1-Year Follow-Up.
BACKGROUND Increased levels of endogenous opioids have been observed in patients with schizophrenia; however, the influence of these endogenous opioids on the biology of schizophrenia remains unclear. The aim of this study was to evaluate the impact of beta-endorphin (BE) on the course of schizophrenia and risk of relapse. MATERIAL AND METHODS The study included 25 patients hospitalized with schizophrenia and 47 controls. Their symptoms were evaluated using Positive and Negative Syndrome Scale (PANSS) and composite index at five points: at the onset of hospitalization; after 4, 6 and 10 weeks of treatment; and after 12 months. ß-endorphin plasma concentrations were assessed in patients at study enrollment and after 6 weeks of treatment. Data regarding rehospitalization during follow-up were also collected. RESULTS Patients had higher BE concentration than controls at study enrollment (P=0.002) and after 6 weeks (P=0.000). BE levels increased during treatment (mean 0.538ng/mL vs. mean 0.624 ng/mL; P=0.007). No correlation was found between BE concentration and PANSS subscale score at any stage of the study. A higher BE level at study enrollment was related to a predominance of negative symptoms after 1 year, measured with composite index (R=-0.404; P=0.045). Patients who were later hospitalized again were significantly more likely to demonstrate an increase in BE levels over 6 weeks (P=0.001). CONCLUSIONS Individuals with schizophrenia demonstrated higher BE concentrations than healthy controls; this tendency was particularly apparent in those affected by negative symptoms. The imbalance in the endogenous opioid system might adversely alter the course of disease and predispose patients to persistence of negative symptoms, despite antipsychotic treatment.


Causes of death among women with breast cancer: A follow-up study of 50 481 women with breast cancer in Finland.
Our study aims to assess mortality from causes other than breast cancer among women with breast cancer with focus on indications of joint aetiology. Data on female breast cancer patients were obtained from the Finnish Cancer Registry and their underlying causes of death in 54 categories from the Statistics Finland. Standardised mortality ratios (SMR) and their 95% confidence intervals (CIs) were calculated for 50 481 patients diagnosed between 1971 and 2000 and followed until December 2012, stratified by histology, age at diagnosis and time since diagnosis. The expected numbers of deaths were based on respective mortality rates among the Finnish general population. Hazard ratio (HR) was estimated from Poisson regression model to compare risks of cause of death by histology. 41% of 30 841 deaths were due to causes other than breast cancer. Significant excess mortality was observed for stomach cancer (SMR 1.43, 95% CI 1.26-1.62), circulatory system diseases (SMR 1.17, 95% CI 1.14-1.20) and suicide (SMR 1.51, 95% CI 1.28-1.78). In an age-adjusted analysis, significantly higher relative risk of stomach cancer mortality was observed for lobular vs ductal subtype (HR 2.00, 95% CI 1.32-3.02). Significantly increased SMRs were observed for cancers of respiratory organs among premenopausal women, and for other respiratory system diseases, dementia and Alzheimer disease among postmenopausal women. We conclude that female breast cancer patients are at increased risk of death from causes other than the breast cancer diagnosis including circulatory and respiratory system diseases and cancer of stomach, ovary and respiratory systems. The excess mortality because of different causes varies based on menopausal status and histology. There might be shared aetiological factors between the diagnosis of breast cancer and the causes of death among these patients.


Influence of clinical and neurocognitive factors in psychosocial functioning after a first episode non-affective psychosis: Differences between males and females.
Deficits in psychosocial functioning are present in the early stages of psychosis. Several factors, such as premorbid adjustment, neurocognitive performance, and cognitive reserve (CR), potentially influence functionality. Sex differences are observed in individuals with psychosis in multiple domains. Nonetheless, few studies have explored the predictive factors of poor functioning according to sex in first-episode psychosis (FEP). This study aimed to explore sex differences, examine changes, and identify predictors of functioning according to sex after onset. The initial sample comprised 588 individuals. However, only adults with non-affective FEP (n = 247, 161 males and 86 females) and healthy controls (n = 224, 142 males and 82 females) were included. A comprehensive assessment including functional, neuropsychological, and clinical scales was performed at baseline and at 2-year follow-up. A linear regression model was used to determine the predictors of functioning at 2-year follow-up. FEP improved their functionality at follow-up (67.4% of both males and females). In males, longer duration of untreated psychosis (β = 0.328, p = 0.003) and worse premorbid adjustment (β = 0.256, p = 0.023) were associated with impaired functioning at 2-year follow-up, while in females processing speed (β = 0.403, p = 0.003), executive function (β = 0.299, p = 0.020) and CR (β = -0.307, p = 0.012) were significantly associated with functioning. Our data indicate that predictors of functioning at 2-year follow-up in the FEP group differ according to sex. Therefore, treatment and preventative efforts may be adjusted taking sex into account. Males may benefit from functional remediation at early stages. Conversely, in females, early interventions centered on CR enhancement and cognitive rehabilitation may be recommended.


Tracking [Formula: see text] of COVID-19: A new real-time estimation using the Kalman filter.
We develop a new method for estimating the effective reproduction number of an infectious disease ([Formula: see text]) and apply it to track the dynamics of COVID-19. The method is based on the fact that in the SIR model, [Formula: see text] is linearly related to the growth rate of the number of infected individuals. This time-varying growth rate is estimated using the Kalman filter from data on new cases. The method is easy to implement in standard statistical software, and it performs well even when the number of infected individuals is imperfectly measured, or the infection does not follow the SIR model. Our estimates of [Formula: see text] for COVID-19 for 124 countries across the world are provided in an interactive online dashboard, and they are used to assess the effectiveness of non-pharmaceutical interventions in a sample of 14 European countries.


Post-radiation valvular and coronary artery disease.
The authors report a case of mitro-aortic valvulopathy associated with single-vessel coronary disease, both probably caused by thoracic irradiation, initially for Hodgkin's lymphoma, and later for carcinoma of the breast. The patient was subjected to aortic valve replacement with a mechanical prosthesis, partial mitral annuloplasty and myocardial revascularization. This report underscores the importance of long-term follow up of patients subjected to mediastinal radiotherapy, because of the significant cardiac morbidity caused by this type of treatment.


Fragile X family with unusual digital and facial abnormalities, cleft lip and palate, and epilepsy.
We present a fragile X family with unusual clinical manifestations. These findings, which often occur in the X-linked FG syndrome, include minor limb anomalies, cleft lip and palate, characteristic facial appearance, gastrointestinal problems and epilepsy, and intellectual disability. In a total sample of 54 fra(X) families, the frequency of minor limb anomalies was estimated to be 32% in the affected males and 19% in the female heterozygotes. These anomalies tend to occur in several members of the same family, where some craniofacial abnormalities reported as characteristic of the FG syndrome have also been encountered. Possible mechanisms for the occurrence of these unusual manifestations in the fra(X) families are discussed.


Human Antimicrobial Peptide Triggered Colloidal Transformations in Bacteria Membrane Lipopolysaccharides.
Growing concerns of bacterial resistance against conventional antibiotics shifts the research focus toward antimicrobial peptide (AMP)-based materials. Most AMPs kill gram-negative bacteria by destroying their inner membrane, but have to first pass the outer membrane covered with lipopolysaccharides (LPS). Their interplay with the LPS is crucial for bactericidal activity, but is yet to be elucidated in detail. In this study, self-assemblies of Escherichia coli LPS with the human cathelicidin AMP LL-37, free and encapsulated into glyceryl monooleate (GMO) lipid nanoparticles, are analyzed using synchrotron small angle X-ray scattering, dynamic light scattering, and cryogenic transmission electron microscopy. Circular dichroism spectroscopy is used to study modifications in LL-37's secondary structure. LPS is found to form elongated micelles and the addition of LL-37 induces their transformation to multilamellar structures. LPS' addition to GMO cubosomes triggers the swelling of the internal cubic structure, while in multilamellar GMO/LL-37 nanocarriers it causes transitions into unstructured particles. The insights on the interactions among LPS and LL-37, in its free form or encapsulated in GMO dispersions, may guide the design of LPS-responsive antimicrobial nanocarriers. The findings may further assist the formulation of antimicrobial nanomaterials with enhanced penetration of LPS layers for improved destruction of bacterial membranes.


Working and strategic memory deficits in schizophrenia.
Working memory and its contribution to performance on strategic memory tests in schizophrenia were studied. Patients (n = 18) and control participants (n = 15), all men, received tests of immediate memory (forward digit span), working memory (listening, computation, and backward digit span), and long-term strategic (free recall, temporal order, and self-ordered pointing) and nonstrategic (recognition) memory. Schizophrenia patients performed worse on all tests. Education, verbal intelligence, and immediate memory capacity did not account for deficits in working memory in schizophrenia patients. Reduced working memory capacity accounted for group differences in strategic memory but not in recognition memory. Working memory impairment may be central to the profile of impaired cognitive performance in schizophrenia and is consistent with hypothesized frontal lobe dysfunction associated with this disease. Additional medial-temporal dysfunction may account for the recognition memory deficit.


Evaluation of a biodegradable graft substitute in rabbit bone defect model.
To evaluate a new biodegradable copolymer calcium sulfate/poly amino acid (CS/PAA) as a graft substitute for the repair of the surgically created cancellous bone defects in rabbits and its biological properties in vivo. Cancellous bone defects were created by drilling holes in the unilateral lateral aspect of the femoral condyle of New Zealand white rabbits. Three groups were assigned: Group A rabbits were grafted with 80% CS/PAA and group B rabbits were grafted with 95% CS/PAA as two treatment groups; group C was sham-operation control group. To study the osteogenic capability in vivo, specimens were harvested at 4, 8, 12, and 16 weeks after implantation and were evaluated by gross assessment, X-ray, histological examination, and histomorphometry. In order to identify the molecular mechanism of bone defect repair, the expression of bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) was detected using Western blot at 4 weeks. Group A and group B showed more vigorous and rapid repair leading to regeneration of cancellous bone than sham-operation control group on gross observation, radiology, and histomorphometry. There was no significant difference between groups A and B. Morphological observation and histological examination showed that the copolymers degraded in sync with the new bone formation process. The expression of BMP-2 and VEGF in implantation groups was higher than that in control group by western blot. These findings demonstrated that the novel biodegradable copolymers can repair large areas of cancellous bone defects. With its controllable degradation rate, it suggests that CS/PAA may be a series of useful therapeutic substitute for bone defects.


Automated two-site immunofluorescent assay for the measurement of serum chromogranin A.
Chromogranin A (CgA) is the best-characterized biological marker common to neuroendocrine tumours and is therefore recommended for their diagnosis. The measurement of serum CgA is of great importance for reaching an early diagnosis and thus reducing the delay before treatment is instigated. The Kryptor CgA assay is the first fully automated assay available. The aim of this study was to evaluate its analytical performance. The imprecision and linearity of the Kryptor CgA assay were evaluated. This assay was compared with the Cis Bio CgA RIA assay in 78 serum samples. Its clinical utility was assessed in serum from 229 patients. The study performed on imprecision of Kryptor measurements showed intra- and inter-run CVs ≤ 5%. The study of linearity showed a satisfactory recovery rate for CgA concentrations up to 1200 μg/L. The Kryptor and RIA assays agreed well on the basis of the cut-off values provided by the two manufacturers. The Bland and Altman plot of the values obtained (range: 20-5560 μg/L) provided a mean difference of -10.1 μg/L (SD: 116). The clinical sensitivities of Kryptor CgA for diagnosis of pheochromocytoma and paraganglioma (n 20) and gastroenteropancreatic NETs (n 17) were respectively 100 and 94%. The Kryptor assay for CgA shows reliable analytical and clinical characteristics and allows a fast delivery of results.


Soil microbiome indicators can predict crop growth response to large-scale inoculation with arbuscular mycorrhizal fungi.
Alternative solutions to mineral fertilizers and pesticides that reduce the environmental impact of agriculture are urgently needed. Arbuscular mycorrhizal fungi (AMF) can enhance plant nutrient uptake and reduce plant stress; yet, large-scale field inoculation trials with AMF are missing, and so far, results remain unpredictable. We conducted on-farm experiments in 54 fields in Switzerland and quantified the effects on maize growth. Growth response to AMF inoculation was highly variable, ranging from -12% to +40%. With few soil parameters and mainly soil microbiome indicators, we could successfully predict 86% of the variation in plant growth response to inoculation. The abundance of pathogenic fungi, rather than nutrient availability, best predicted (33%) AMF inoculation success. Our results indicate that soil microbiome indicators offer a sustainable biotechnological perspective to predict inoculation success at the beginning of the growing season. This predictability increases the profitability of microbiome engineering as a tool for sustainable agricultural management.


Cross-coupling between insulin and estrogen receptor in human neuroblastoma cells.
Insulin is a well known mitotic agent for neuroblastoma cells. Human SK-N-BE neuroblastoma cells stably transfected with the estrogen receptor, however, undergo growth arrest and differentiation when treated with insulin. These effects were shown to be due to an insulin-dependent activation of the unliganded estrogen receptor. Here, we demonstrate that this activation involves the AF-2 COOH-terminal domain of the estrogen receptor and that the communication between estrogen and insulin receptor systems occurs via selected and specific transduction signals. In fact, by the use of dominant negative and dominant positive mutants we demonstrate that p21ras is essential for insulin and estrogen receptor coupling. With pharmacological tools, we prove that PI 3'kinase does not contribute to this cross-talk and that protein kinase C triggers transduction signals that act in synergism with p21ras. These results prove the intricacy of all these intracellular paths of communication. The finding that, in neuroblastoma cells, selected signal transduction systems are involved in the insulin-dependent activation of estrogen receptor is of particular interest considering that estrogen receptor might restrict the role played by insulin during the differentiation of neural cells and interfere with its proliferative potential while allowing its regulation of other functions related to cell survival.


Purification and properties of a cellulase from Aspergillus niger.
A cellulolytic enzyme was isolated from a commercial cellulase preparation form Aspergillus niger. A yield of about 50mg of enzyme was obtained per 100g of commerial cellulase. The isolated enzyme was homogeneous in the ultracentrifuge at pH 4.0 and 8.0, and in sodium dodecyl sulphate/polyacrylamide-gel electrophoresis but showed one major and two minor bands in disc gel electrophoresis. No carbohydrate was associated with the protein. Amino acid analysis revealed that the enzyme was rich in acidic and aromatic amino acids. Data from the amino acid composition and dodecyl sulphate/polyacrylamide-gel electrophoresis indicated a molecular weight of 26000. The purified enzyme was active towards CM-cellulose, but no activity towards either cellobiose or p-nitrophenyl beta-D-glucoside was detected under the assay conditions used. The pH optimum for the enzyme was pH 3.8-4.0, and it was stable at 25 degrees C over the range pH 1-9; maximum activity (at pH 4.0) was obtained at 45 degrees C. The cellulase was more stable to heat treatment at pH 8.0 than at 4.0. Kinetic studies gave pK values between 4.2 and 5.3 for groups involved in the enzyme-substrate complex.


A compartmental model for simulation of IGF-I kinetics and metabolism.
A compartmental model for the simulation of in-vivo insulin-like growth factor one (IGF-I) kinetics and metabolism is presented. The model is based on the concept that IGF binding proteins (IGFBPs) act as a reservoir for IGF-I in plasma, thereby restricting its access to the extravascular space. The model is divided into two subsystems; a binding model for IGF-I and IGFBP, and an IGF-I clearance model, which includes the binding of IGF-I to IGF-I receptors. The non-linear differential equations describing the model are based on the law of mass action. Given the steady state condition, the linearized form of the model, describing tracer kinetics of IGF-I and IGFBPs is developed. It is demonstrated that using the available data, it is possible to assign values to the model parameters, thereby allowing simulation of the model to obtain information not accessible by direct observation.


Characterizing the expression of the human olfactory receptor gene family using a novel DNA microarray.
Olfactory receptor (OR) genes were discovered more than a decade ago, when Buck and Axel observed that, in rats, certain G-protein coupled receptors are expressed exclusively in the olfactory epithelium. Subsequently, protein sequence similarity was used to identify entire OR gene repertoires of a number of mammalian species, but only in mouse were these predictions followed up by expression studies in olfactory epithelium. To rectify this, we have developed a DNA microarray that contains probes for most predicted human OR loci and used that array to examine OR gene expression profiles in olfactory epithelium tissues from three individuals. We detected expression of 437 (76%) human OR genes in these olfactory epithelia. Interestingly, we detected widespread expression of OR pseudogenes, an observation that may shed light on the mechanism of OR gene choice in the olfactory sensory neurons. To address the hypothesis that OR genes may carry out additional functions, we also characterized the expression of OR genes in a number of non-olfactory tissues. While our results corroborate the functional annotation of the majority of predicted human odorant receptors, we find that a large number of putative human OR genes are expressed in non-olfactory tissues, sometimes exclusively so. Our evolutionary analysis of ectopically expressed human OR genes does not lend support to the hypothesis that these genes have alternative functions.


Transient receptor potential canonical type 3 channels facilitate endothelium-derived hyperpolarization-mediated resistance artery vasodilator activity.
Microdomain signalling mechanisms underlie key aspects of artery function and the modulation of intracellular calcium, with transient receptor potential (TRP) channels playing an integral role. This study determines the distribution and role of TRP canonical type 3 (C3) channels in the control of endothelium-derived hyperpolarization (EDH)-mediated vasodilator tone in rat mesenteric artery. TRPC3 antibody specificity was verified using rat tissue, human embryonic kidney (HEK)-293 cells stably transfected with mouse TRPC3 cDNA, and TRPC3 knock-out (KO) mouse tissue using western blotting and confocal and ultrastructural immunohistochemistry. TRPC3-Pyr3 (ethyl-1-(4-(2,3,3-trichloroacrylamide)phenyl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate) specificity was verified using patch clamp of mouse mesenteric artery endothelial and TRPC3-transfected HEK cells, and TRPC3 KO and wild-type mouse aortic endothelial cell calcium imaging and mesenteric artery pressure myography. TRPC3 distribution, expression, and role in EDH-mediated function were examined in rat mesenteric artery using immunohistochemistry and western blotting, and pressure myography and endothelial cell membrane potential recordings. In rat mesenteric artery, TRPC3 was diffusely distributed in the endothelium, with approximately five-fold higher expression at potential myoendothelial microdomain contact sites, and immunoelectron microscopy confirmed TRPC3 at these sites. Western blotting and endothelial damage confirmed primary endothelial TRPC3 expression. In rat mesenteric artery endothelial cells, Pyr3 inhibited hyperpolarization generation, and with individual SK(Ca) (apamin) or IK(Ca) (TRAM-34) block, Pyr3 abolished the residual respective IK(Ca)- and SK(Ca)-dependent EDH-mediated vasodilation. The spatial localization of TRPC3 and associated channels, receptors, and calcium stores are integral for myoendothelial microdomain function. TRPC3 facilitates endothelial SK(Ca) and IK(Ca) activation, as key components of EDH-mediated vasodilator activity and for regulating mesenteric artery tone.


Regulation of sodium transport in M-1 cells.
The M-1 cell line, derived from the mouse cortical collecting duct (CCD), is being used as a mammalian model of the CCD to study Na+ transport. The present studies aimed to further define the role of various hormones in affecting Na+ transport in M-1 cells grown in defined media. M-1 cells on permeable support, in serum-free media, developed amiloride-sensitive current 4-5 days after seeding. As expected for the involvement of epithelial Na+ channels, alpha-, beta-, and gamma-subunits of the epithelial Na+ channel were identified by RT-PCR. Either dexamethasone (Dex, 10-100 nM) or aldosterone (Aldo, 10(-6)-10(-7) M) for 24 h stimulated transport. Cells grown in the presence of Aldo and Dex had higher transport than with Dex alone. Spironolactone added to Dex media decreased transport. The acute effects of hormones reported to inhibit Na+ transport in CCD were also examined. Epidermal growth factor, phorbol esters, and increased intracellular Ca2+ with thapsigargin did not alter transport. Arginine vasopressin caused a transient increase in transport (probably Cl- secretion), which was not amiloride sensitive. Also, the protease inhibitor aprotinin decreased Na+ transport; in aprotinin-treated cells, trypsin stimulated transport. This study demonstrates that adrenal steroids (Dex > Aldo) stimulate Na+ transport in M-1 cells. At least part of this response may represent activation of mineralocorticoid receptors based on an additive effect of Dex and Aldo, as well as inhibition by spironolactone. Responses to immediate-acting hormones is limited. However, an endogenous protease activity, which activates Na+ transport, is present in these cells.


Roles of tissue-resident immune cells in immunotherapy of non-small cell lung cancer.
Non-small cell lung cancer (NSCLC) is the most common and lethal type of lung cancer, with limited treatment options and poor prognosis. Immunotherapy offers hope for improving the survival and quality of life of NSCLC patients, but its efficacy depends on the tumor immune microenvironment (TME). Tissue-resident immune cells are a subset of immune cells that reside in various tissues and organs, and play an important role in fighting tumors. In NSCLC, tissue-resident immune cells are heterogeneous in their distribution, phenotype, and function, and can either promote or inhibit tumor progression and response to immunotherapy. In this review, we summarize the current understanding on the characteristics, interactions, and roles of tissue-resident immune cells in NSCLC. We also discuss the potential applications of tissue-resident immune cells in NSCLC immunotherapy, including immune checkpoint inhibitors (ICIs), other immunomodulatory agents, and personalized cell-based therapies. We highlight the challenges and opportunities for developing targeted therapies for tissue-resident immune cells and optimizing existing immunotherapeutic approaches for NSCLC patients. We propose that tissue-resident immune cells are a key determinant of NSCLC outcome and immunotherapy response, and warrant further investigation in future research.


A parapoxviral virion protein targets the retinoblastoma protein to inhibit NF-κB signaling.
Poxviruses have evolved multiple strategies to subvert signaling by Nuclear Factor κB (NF-κB), a crucial regulator of host innate immune responses. Here, we describe an orf virus (ORFV) virion-associated protein, ORFV119, which inhibits NF-κB signaling very early in infection (≤ 30 min post infection). ORFV119 NF-κB inhibitory activity was found unimpaired upon translation inhibition, suggesting that virion ORFV119 alone is responsible for early interference in signaling. A C-terminal LxCxE motif in ORFV119 enabled the protein to interact with the retinoblastoma protein (pRb) a multifunctional protein best known for its tumor suppressor activity. Notably, experiments using a recombinant virus containing an ORFV119 mutation which abrogates its interaction with pRb together with experiments performed in cells lacking or with reduced pRb levels indicate that ORFV119 mediated inhibition of NF-κB signaling is largely pRb dependent. ORFV119 was shown to inhibit IKK complex activation early in infection. Consistent with IKK inhibition, ORFV119 also interacted with TNF receptor associated factor 2 (TRAF2), an adaptor protein recruited to signaling complexes upstream of IKK in infected cells. ORFV119-TRAF2 interaction was enhanced in the presence of pRb, suggesting that ORFV119-pRb complex is required for efficient interaction with TRAF2. Additionally, transient expression of ORFV119 in uninfected cells was sufficient to inhibit TNFα-induced IKK activation and NF-κB signaling, indicating that no other viral proteins are required for the effect. Infection of sheep with ORFV lacking the ORFV119 gene led to attenuated disease phenotype, indicating that ORFV119 contributes to virulence in the natural host. ORFV119 represents the first poxviral protein to interfere with NF-κB signaling through interaction with pRb.


Disease-Associated Mutation A554V Disrupts Normal Autoinhibition of DNMT1.
DNA methyltransferase 1 (DNMT1) is the enzyme primarily responsible for propagation of the methylation pattern in cells. Mutations in DNMT1 have been linked to the development of adult-onset neurodegenerative disorders; these disease-associated mutations occur in the regulatory replication foci-targeting sequence (RFTS) domain of the protein. The RFTS domain is an endogenous inhibitor of DNMT1 activity that binds to the active site and prevents DNA binding. Here, we examine the impact of the disease-associated mutation A554V on normal RFTS-mediated inhibition of DNMT1. Wild-type and mutant proteins were expressed and purified to homogeneity for biochemical characterization. The mutation increased DNA binding affinity ~8-fold. In addition, the mutant enzyme exhibited increased DNA methylation activity. Circular dichroism (CD) spectroscopy revealed that the mutation does not significantly impact the secondary structure or relative thermal stability of the isolated RFTS domain. However, the mutation resulted in changes in the CD spectrum in the context of the larger protein; a decrease in relative thermal stability was also observed. Collectively, this evidence suggests that A554V disrupts normal RFTS-mediated autoinhibition of DNMT1, resulting in a hyperactive mutant enzyme. While the disease-associated mutation does not significantly impact the isolated RFTS domain, the mutation results in a weakening of the interdomain stabilizing interactions generating a more open, active conformation of DNMT1. Hyperactive mutant DNMT1 could be responsible for the increased DNA methylation observed in affected individuals.


Perfluorooctanoic acid exposure is associated with elevated homocysteine and hypertension in US adults.
To investigate the association between serum perfluorooctanoic acid (PFOA) concentration and cardiovascular disease, as measured by homocysteine level and blood pressure in a representative sample of US adults. A cross-sectional study of 2934 adults (≥20 years) who participated in the 2003-2004 and 2005-2006 National Health and Nutrition Examination Survey and had detectable levels of PFOA in their serum. The health effects analysed as potentially associated with PFOA exposure included homocysteine level and blood pressure. The geometric mean value (95% CI) of the study participants' serum PFOA concentration was 4.00 μg/l (95% CI 3.86 to 4.13). The homocysteine and systolic blood pressure were shown to increase significantly with an increase in the log-transformed serum PFOA concentration, after adjusting for potential confounding variables. Adjusted ORs comparing participants at the 80th versus the 20th percentiles were 2.62 for hypertension (95% CI 2.09 to 3.14), and a positive association was also evident in models based on quartiles or based on restricted cubic splines. These findings suggest that background exposure to PFOA may continue a risk factor for the development of cardiovascular diseases.


Persistent, consistent, widespread, and robust? Another look at recent trends in old-age disability.
The purpose of this study was to provide new evidence on disability trends among elderly persons from 1982 to 1996. The sample includes 124,949 participants aged 70 and older in the 1982-1996 National Health Interview Surveys. Logistic analysis was used to estimate the trend in disability prevalence after controlling for various sociodemographic factors. We found that: (a) the prevalence of disability has declined, but the gains did not persist throughout the entire period or accelerate over time; (b) only routine care disability has declined, whereas more severe personal care disability shows no improvements; (c) estimates are robust to the exclusion of the nursing home population but may be sensitive to growth in the assisted living population; (d) estimates of decline in disability prevalence are fairly consistent across five national surveys; (e) gains have been concentrated among the most educated elderly persons; and (f) gains in education appear to be an important confounder of the improvements. Evidence from several surveys using various measures indicates that disability has declined among elderly persons. Determining the causes of the improvements should be a high priority in future research efforts.


Galectin-3 Participates in Cardiovascular Remodeling Associated With Obesity.
Remodeling, diastolic dysfunction, and arterial stiffness are some of the alterations through which obesity affects the cardiovascular system. Fibrosis and inflammation are important mechanisms underlying cardiovascular remodeling, although the precise promoters involved in these processes are still unclear. Galectin-3 (Gal-3) induces inflammation and fibrosis in the cardiovascular system. We have investigated the potential role of Gal-3 in cardiac damage in morbidly obese patients, and we have evaluated the protective effect of the Gal-3 inhibition in the occurrence of cardiovascular fibrosis and inflammation in an experimental model of obesity. Morbid obesity is associated with alterations in cardiac remodeling, mainly left ventricular hypertrophy and diastolic dysfunction. Obesity and hypertension are the main determinants of left ventricular hypertrophy. Insulin resistance, left ventricular hypertrophy, and circulating levels of C-reactive protein and Gal-3 are associated with a worsening of diastolic function in morbidly obese patients. Obesity upregulates Gal-3 production in the cardiovascular system in a normotensive animal model of diet-induced obesity by feeding for 6 weeks a high-fat diet (33.5% fat). Gal-3 inhibition with modified citrus pectin (100 mg/kg per day) reduced cardiovascular levels of Gal-3, total collagen, collagen I, transforming and connective growth factors, osteopontin, and monocyte chemoattractant protein-1 in the heart and aorta of obese animals without changes in body weight or blood pressure. In morbidly obese patients, Gal-3 levels are associated with diastolic dysfunction. In obese animals, Gal-3 blockade decreases cardiovascular fibrosis and inflammation. These data suggest that Gal-3 could be a novel therapeutic target in cardiac fibrosis and inflammation associated with obesity.


Injection Molded Capsules for Colon Delivery Combining Time-Controlled and Enzyme-Triggered Approaches.
A new type of colon targeting system is presented, combining time-controlled and enzyme-triggered approaches. Empty capsule shells were prepared by injection molding of blends of a high-amylose starch and hydroxypropyl methylcellulose (HPMC) of different chain lengths. The dissolution/erosion of the HPMC network assures a time-controlled drug release, i.e., drug release starts upon sufficient shell swelling/dissolution/erosion. In addition, the presence of high-amylose starch ensures enzyme-triggered drug release. Once the colon is reached, the local highly concentrated bacterial enzymes effectively degrade this polysaccharide, resulting in accelerated drug release. Importantly, the concentration of bacterial enzymes is much lower in the upper gastrointestinal tract, thus enabling site-specific drug delivery. The proposed capsules were filled with acetaminophen and exposed to several aqueous media, simulating the contents of the gastrointestinal tract using different experimental setups. Importantly, drug release was pulsatile and occurred much faster in the presence of fecal samples from patients. The respective lag times were reduced and the release rates increased once the drug started to be released. It can be expected that variations in the device design (e.g., polymer blend ratio, capsule shell geometry and thickness) allow for a large variety of possible colon targeting release profiles.


Social-emotional processing in nonverbal individuals with Angelman syndrome: evidence from brain responses to known and novel names.
The combination of intellectual, communicative and motor deficits limits the use of standardised behavioural assessments in individuals with Angelman syndrome (AS). The current study aimed to objectively evaluate the extent of social-emotional processing in AS using auditory event-related potentials (ERPs) during passive exposure to spoken stimuli. Auditory ERP responses were recorded in 13 nonverbal individuals with the deletion subtype of AS, age 4-45 years, during the name recognition paradigm, in which their own names and names of close others (relative or friend) were presented among novel names. No behavioural responses were required. Contrary to findings in typical children and adults, there was no significant evidence of differential neural response to known vs. novel names in participants with AS. Nevertheless, greater amplitude differences between known and unknown names demonstrated the predicted association with better interpersonal relationships and receptive communication abilities. These findings indicate good tolerability of ERP procedures (85% success rate). The lack of own name differentiation is consistent with increased incidence of the autism-related symptoms in AS. Strong associations between the caregiver reports of adaptive functioning and neural indices of known name recognition support the utility of brain-based measures for objectively evaluating cognitive and affective processes in nonverbal persons with neurodevelopmental disorders.


[What has spinal stabilization contributed for the paraplegic patient?].
The aim of spondylodesis is to restore the injured spinal column to its previous shape and stability. In the transverse SCI patient, it entails easier intensive care in case of multiple injury as well as easier general nursing tasks, and greater intensity of physiotherapy in the acute and early phases. Compared to conservative management of the injured spinal column, mobilization is effected some 21 to 39 days earlier in surgical patients, total hospitalization in tetraplegics is shortened by 5 days, and by 34 to 57 days in paraplegic patients. Functionally valuable neurological recovery in primary complete paralysis occurs in 19%/20% of conservative/surgical cases, in primary partial paralysis in 67%/58%. Stabilization of the spinal column however does not at the same time achieve a stabilization of the transverse SCI patient's entire situation--physical, psychological, social. Irrespective of the scope of paralysis, the advantages of a spondylodesis should be considered also in case of total paraplegia. Conservative management retains its importance in cases where surgery is not indicated, or even contraindicated. Comparable results are very rarely achieved outside the specialized centres. Acute cases of SCI should therefore be referred to spinal centres as soon as possible, which for their part have to be able to admit patients 24 hours a day, as well as to provide adequate care until completion of the entire treatment.


[Effect of psychotropic substances on dopamine uptake by synaptosomes and glial cells].
Dophamine-H3 uptake by synaptosomes and glial cells of the cerebral cortex in rabbits and by the synaptosomes of the cerebral cortex in rats was studied. KM value of the dophamine-H3 uptake was found to be the same (0.075 +/- +/- 0.01 micrometer) in case of synaptosomes and glial cells. The rate of dophamine-H3 uptake by the rabbit cerebral cortex synaptosomes was twice that of the gial cells (as compared by protein). Among the psychotropic agents studied the most active symaptosome uptake inhibitors were phenamine and cocaine; they were less active in respect to glial uptake. Both types of uptake were inhibited by neuroleptics and antidepressants to the same degree. Results of this work conformed to the view on the participation of the strial dophaminergic link in the action mechanism of psychostimulants.


Left ventricular thrombi identified by cross-sectional echocardiography.
We studied 25 patients with anterior myocardial infarction and two with congestive cardiomyopathy to evaluate two-dimensional echocardiography in the diagnosis of left ventricular thrombi. Five coronary patients had systemic emboli. Four of these patients manifested apical filling defects on cineangiogram, while a levophase cine was equivocal for clot in the fifth patient. Neither echocardiography nor cineangiography visualized ventricular thrombi in the nonembolus coronary patients. Echoes from a distinct apical mass, however, were visualized in all five patients in the embolus group by two-dimensional echocardiography at the cardiac apex. Apical thrombi were confirmed in all four patients in the embolus group undergoing surgery. The irregular configuration of recent thrombi in the coronary patients differed from the circumscribed appearance of chronic thrombi in the cardiomyopathy patients on two-dimensional echocardiogram. Thus, two-dimensional echocardiography can be used to detect and characterize left ventricular thrombi.


AtAIRP2 E3 Ligase Affects ABA and High-Salinity Responses by Stimulating Its ATP1/SDIRIP1 Substrate Turnover.
AtAIRP2 is a cytosolic RING-type E3 ubiquitin ligase that positively regulates an abscisic acid (ABA) response in Arabidopsis (Arabidopsis thaliana). Yeast two-hybrid screening using AtAIRP2 as bait identified ATP1 (AtAIRP2 Target Protein1) as a substrate of AtAIRP2. ATP1 was found to be identical to SDIRIP1, which was reported recently to be a negative factor in ABA signaling and a target protein of the RING E3 ligase SDIR1. Accordingly, ATP1 was renamed ATP1/SDIRIP1. A specific interaction between AtAIRP2 and ATP1/SDIRIP1 and ubiquitination of ATP1/SDIRIP1 by AtAIRP2 were demonstrated in vitro and in planta. The turnover of ATP1/SDIRIP1 was regulated by AtAIRP2 in cell-free degradation and protoplast cotransfection assays. The ABA-mediated germination assay of 35S:ATP1/SDIRIP1-RNAi/atairp2 double mutant progeny revealed that ATP1/SDIRIP1 acts downstream of AtAIRP2. AtAIRP2 and SDIR1 reciprocally complemented the ABA- and salt-insensitive germination phenotypes of sdir1 and atairp2 mutants, respectively, indicating their combinatory roles in seed germination. Subcellular localization and bimolecular fluorescence complementation experiments in the presence of MG132, a 26S proteasome inhibitor, showed that AtAIRP2 and ATP1/SDIRIP1 were colocalized to the cytosolic spherical body, which lies in close proximity to the nucleus, in tobacco (Nicotiana benthamiana) leaf cells. The 26S proteasome subunits RPN12a and RPT1 and the molecular chaperones HSP70 and HSP101 were colocalized to these discrete punctae-like structures. These results raised the possibility that AtAIRP2 and ATP1/SDIRIP1 interact in the cytosolic spherical compartment. Collectively, our data suggest that the down-regulation of ATP1/SDIRIP1 by AtAIRP2 and SDIR1 RING E3 ubiquitin ligases is critical for ABA and high-salinity responses during germination in Arabidopsis.


Strong environmental determination of a carotenoid-based plumage trait is not mediated by carotenoid availability.
Carotenoid-based colours are recognized as having an important signalling function, yet the nature of the mechanisms that maintain their honesty is not well understood. By combining a carotenoid-feeding experiment with a quantitative genetic experiment in a wild population of blue tits (Parus caeruleus), we were able to test predictions that differentiate between proposed mechanisms. If variation in carotenoid ingestion underlies variation in carotenoid-based colour expression, then carotenoid-supplemented birds should have reduced variance in colour. In this study, carotenoid supplementation produced a small but significant change in plumage colouration, but no significant change in variation. These results suggest that variation in carotenoid acquisition is not an important source of variation for this colour trait, and that variation in post-ingestion processes are likely to be more important. The low heritability of this colour trait suggests environmental factors are likely to underlie the majority of variation in these processes.


[A checklists' jungle].
The steps of any procedure can be summed up in a checklist. The question is whether that is efficient and effective. A patient safety checklist must be more than a collection of tick boxes. An effective safety checklist in healthcare should cover only complex procedures with an inherent risk hazard, being performed in a complex environment, where memory and situational awareness of a team can be channelled by using a checklist. It is counterproductive to have separate checklists, each covering an isolated part of a process. The use of technical equipment by medical personnel is prone to error. Knowledge of how to use that piece of equipment is essential; medical specialists have their own responsibility in terms of possession of adequate skills. Maintenance and function checks are key to safe use. Checks of equipment or devices prior to surgery should be done as part of a generic perioperative checklist.


Family factors in the development and management of anxiety disorders.
Family variables are thought to play a key role in a wide variety of psychopathology according to many theories. Yet, specific models of the development of anxiety disorders place little emphasis on general family factors despite clear evidence that anxiety runs in families. The current review examines evidence for the involvement of a number of family-related variables in the development of anxiety disorders as well as the importance of families in their management. Evidence across most areas is shown to be weak and inconsistent, with the one exception being an extensive literature on the role of parenting in the development of anxiety. There is also currently little evidence that family factors have a strong role to play in the treatment of anxiety, aside from research demonstrating the value of parents and partners as non-critical supports in therapy. The promises and hints in the literature, combined with the currently inconsistent methods, suggest that considerably more research is needed to determine whether specific family factors may yet be shown to play a key role in the development and management of anxiety disorders.


[Release of Ca2+ ions from the sarcoplasmic reticulum of skeletal muscles after treatment with caffeine].
Using a Ca2+-selective electrode and Quin 2 and chlortetracycline fluorescence spectra, a comparative study of caffeine- and Ca2+-induced release of Ca2+ from the terminal cisterns of rabbit fast skeletal muscle sarcoplasmic reticulum was carried out. It was shown that the caffeine-induced release of Ca2+ depends on Ca2+ and Mg2+ concentration in the medium; Mg2+ inhibit, while Ca2+ stimulate this process. The caffeine-induced transport of Ca2+ is blocked by ruthenium red, tetracaine and dimethylsulfoxide. The Ca2+ release induced by Ca2+ was shown to occur in two ways, i. e., via Mg2+-dependent (inhibited by Mg2+ and caffeine blockers) and Mg2+-independent (insensitive to caffeine inhibitors, including Mg2+) routes. It was assumed that caffeine stimulates the Mg2+-dependent, Ca2+-induced release of Ca2+. The sensitivity of Ca2+ transport to caffeine testifies to the fact that about 80% of the total Ca2+ transport activity of fast skeletal muscle homogenates belongs to terminal cisterns. The total amount of sarcoplasmic reticulum membranes in the muscle makes up to 15-20 mg of protein/g of tissue.


Separate and combined effects of neurotoxic and lytic compounds of Alexandrium strains on Mytilus edulis feeding activity and hemocyte function.
Multiple toxic and bioactive compounds produced by Alexandrium spp. cause adverse effects on bivalves, but these effects are frequently difficult to attribute to a single compound class. To disentangle the effect of neurotoxic vs lytic secondary metabolites, we exposed blue mussels to either a paralytic shellfish toxin (PST) producing Alexandrium spp. strain, or to an exclusively lytic compound (LC) producing strain, or a strain containing both compound classes, to evaluate the time dependent effects after 3 and 7 days of feeding. Tested parameters comprised signs of paralysis, feeding activity, and immune cell integrity (hemocyte numbers and viability; lysosomal membrane destabilization) and function (ROS production). Both compound classes caused paralysis and immune impairment. The only effect attributable exclusively to PST was increased phagocytic activity after 3 days and impaired feeding activity after 7 days, which curtailed toxin accumulation in digestive glands. Lysosomal membrane destabilization were more closely, but not exclusively, matched with LC exposure. Effects on circulating hemocyte integrity and immune related functions were mostly transient or remained stable within 7 days; except for increased lysosomal labialization and decreased extracellular ROS production when mussels were exposed to the toxin combination. M. edulis displays adaptive fitness traits to survive and maintain immune capacity upon prolonged exposure to environmentally relevant concentrations of PST and/or LC producing Alexandrium strains.


Diffractive distortion of a pixelated computer-generated hologram with oblique illumination.
Computer-generated holograms (CGHs) have their phase and/or amplitude modulation pattern calculated rather than recorded as for traditional holograms. In practice, the CGH devices are normally pixelated, no matter if they are passive or active ones. In many cases, the reconstruction light illuminates on the CGH devices obliquely, and the pattern generated on the target plane will be distorted from the originally desired one, even if the modulation on the CGH devices has been calibrated for the corresponding illumination angle in CGH calculation and optimization. The distortion is purely related to the diffraction behavior resulting from the geometry of the pixel on the CGH, and therefore diffractive distortion has been coined for this specific phenomenon. In this paper, quantitative analysis of diffractive distortion and a corresponding scheme for correction have been given based on scalar diffraction theory. The proposed concept is that the distortion of the reconstructed image is proportional to the distortion of the signal window. An experiment has been conducted with a phase-type liquid crystal on silicon spatial light modulator (SLM). Both the distortion analysis and the correction scheme have been verified quantitatively for various illumination angles and the direction of the reconstruction light.


PET/MR imaging: technical aspects and potential clinical applications.
Instruments that combine positron emission tomography (PET) and magnetic resonance (MR) imaging have recently been assembled for use in humans, and may have diagnostic performance superior to that of PET/computed tomography (CT) for particular clinical and research applications. MR imaging has major strengths compared with CT, including superior soft-tissue contrast resolution, multiplanar image acquisition, and functional imaging capability through specialized techniques such as diffusion-tensor imaging, diffusion-weighted (DW) imaging, functional MR imaging, MR elastography, MR spectroscopy, perfusion-weighted imaging, MR imaging with very short echo times, and the availability of some targeted MR imaging contrast agents. Furthermore, the lack of ionizing radiation from MR imaging is highly appealing, particularly when pediatric, young adult, or pregnant patients are to be imaged, and the safety profile of MR imaging contrast agents compares very favorably with iodinated CT contrast agents. MR imaging also can be used to guide PET image reconstruction, partial volume correction, and motion compensation for more accurate disease quantification and can improve anatomic localization of sites of radiotracer uptake, improve diagnostic performance, and provide for comprehensive regional and global structural, functional, and molecular assessment of various clinical disorders. In this review, we discuss the historical development, software-based registration, instrumentation and design, quantification issues, potential clinical applications, potential clinical roles of image segmentation and global disease assessment, and challenges related to PET/MR imaging. http://radiology.rsna.org/lookup/suppl/doi:10.1148/radiol.13121038/-/DC1.


Pharmacy on-site overdose protocols and prevention of overdose.
Opioid overdose is a preventable injury leading to high morbidity and premature mortality in communities across the United States. Overdoses take place where people use drugs, including commercial and public locations like community pharmacies, and necessitate swift detection and response to avoid harm and, even more seriously, death. The presence of emergency and safety protocols improves occupational health and safety for all in the workplace. The aim of this study was to assess the prevalence of experience with on-site pharmacy overdose and to explore pharmacist and site characteristics associated with having a known protocol for responding to on-site overdose emergencies. Methods: An anonymous, online survey about naloxone provision and opioid safety was delivered by email, through professional pharmacy associations and continuing education attendance lists, to 3,100 pharmacists in Massachusetts and Rhode Island between October 2017 and January 2018. Survey items gauged socio-demographics, practice site characteristics, safer opioid dispensing and naloxone provision. Summary statistics and bivariate analyses were conducted to describe characteristics associated with items pertaining to on-site overdose policy awareness. Results: Of the 357 respondents (11.5% response rate), 154 (5.0%) answered the questions of interest: 17.5% reported having at least one suspected overdose on-site at their practice location, while 42.9% reported that they were knowledgeable about and could locate at their practice location an on-site overdose protocol detailing how to respond to an overdose. Pharmacists who were knowledgeable about protocols were also more likely to offer naloxone to patients (p = 0.02) and did not practice at a chain pharmacy (p = 0.01). Conclusions: Community pharmacies that stock and distribute naloxone are key parts of community efforts to address the opioid crisis. Pharmacies and other healthcare settings should develop and implement on-site overdose response protocols and cultivate a norm of naloxone provision to patients.


An I/O device driver for bioinformatics tools: the case for BLAST.
There are many bioinformatics tools that deal with input/output (I/O) issues by using filing systems from the most common operating systems, such as Linux or MS Windows. However, as data volumes increase, there is a need for more efficient disk access, ad hoc memory management and specific page-replacement policies. We propose a device driver that can be used by multiple applications. It keeps the application code unchanged, providing a non-intrusive and flexible strategy for I/O calls that may be adopted in a straightforward manner. With our approach, database developers can define their own I/O management strategies. We used our device driver to manage Basic Local Alignment Search Tool (BLAST) I/O calls. Based on preliminary experimental results with National Center for Biotechnology Information (NCBI) BLAST, this approach can provide database management systems-like data management features, which may be used for BLAST and many other computational biology applications.


High dose, dose-intensive chemotherapy with doxorubicin and cyclophosphamide for the treatment of advanced breast cancer.
Eighteen patients with advanced breast cancer were commenced on treatment with high dose doxorubicin (100 mg m-2) or doxorubicin (100 mg m-2) and cyclophosphamide (500 mg m-2) at 2 weekly intervals. Three cycles of treatment were planned. rG-CSF was given subcutaneously for 10 days, starting 24 h after each cycle of chemotherapy. Sixteen out of 18 patients responded (89%) of whom six (33%) achieved a complete remission. Twelve (67%) completed the three planned cycles, four (22%) received two cycles and two (11%) received one cycle only. The median time to progression was 5 1/2 months and the median survival was 18 1/2 months. Neutropenia occurred after 89% of courses and 65% of courses were accompanied by a significant (WHO grade III or IV) infection. The duration of neutropenia was short (mean 5.4 days) and mean time to absolute neutrophil count recovery (ANC > 1,000 x 10(6) litre) from the start of treatment was 11 days. Moderate to severe epithelial toxicity (WHO grade 3 or 4) accompanied 43% of courses and was dose limiting. High dose, dose intensive chemotherapy has an excellent initial therapeutic effect in advanced breast cancer but does not prolong duration of remission or overall survival beyond that of standard treatment. Although subcutaneous rG-CSF curtailed the expected duration of neutropenia substantially, the overall incidence of neutropenia and of infections requiring intravenous antibiotics was high. Furthermore, almost half of the courses were complicated by moderate to severe oral mucositis and/or mild to moderate palmar and plantar inflammation. The lack of survival benefit and excess toxicity seriously limits the wider application of this regime. It should not be used in place of standard dose palliative chemotherapy for metastatic breast cancer.


Structural specificity in the renal tubular transport of tyrosine.
The structural requirements for aromatic amino acid reabsorption and secretion in the dog were investigated using a series of tyrosine analogs. In clearance experiments, l-tyrosine and l-phenylalanine underwent net reabsorption at rates 5 to 6 times greater than those of the corresponding d-enantiomers and about 1.5 time those of o- and m-dl-tyrosine. 3-(p-hydroxyphenyl)Propionic acid underwent net secretion under conditions of polyuria and high urinary pH and net reabsorption under conditions of oliguria and low pH. In stop-flow experiments, (U/Pl-amino acid)/(U/Pln) ratios, where U/P = concentration in urine and plasma and In = inulin, of proximal tubular samples were 1.5 to 5-fold greater than the control (free-flow) values of 0.1 to 0.3, indicating tubular secretion. Secretion was inhibited, and net reabsorption enhanced, by probenecid, p-aminohippuric acid and 2,4-dinitrophenol. (U/Pd-amino acid)/(U/PIN) ratios of proximal tubular samples were below the control values of 0.6 to 0.8, indicating tubular reabsorption. Probenecid had no effect on the overall pattern but caused a slight decrease in the relative clearance values. The stop-flow pattern of 3-(p-hydroxyphenyl)propionic acid excretion was similar to that of the I-amino acids. It was concluded that the I-amino acids undergo bidirectional transport and that the I-configuration and amino group are required for optimal active reabsorption, whereas ring hydroxylation has little effect on reabsorption transport. No absolute structural requirements for active secretion were elucidated.


Dendrimer crosslinked collagen as a corneal tissue engineering scaffold: mechanical properties and corneal epithelial cell interactions.
Generation 2 polypropyleneimine octaamine dendrimers were used to generate highly crosslinked collagen with mechanical properties that would make it appropriate for use as a corneal tissue-engineering scaffold. Crosslinking of a highly concentrated collagen solution (2-4%) was effected using the water-soluble carbodiimide 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC). The multifunctional dendrimers were introduced as novel multifunctional crosslinkers after the activation of the carboxylic acid groups of glutamic and aspartic acid residues in collagen. Glutaraldehyde, a common collagen crosslinker, was used as comparison, as was EDC, itself an alternative crosslinker, which forms "zero-length or self-crosslinking". The mechanical properties resultant gels were determined. Young's modulus of the dendrimer crosslinked gels was significantly higher than that observed with the other crosslinkers, increasing to 5 MPa compared with 0.1 MPa for the EDC crosslinked gels. Transmission electron microscopy (TEM) analysis of the gels demonstrated the presence of fibrils in the thermally gelled collagen controls; no fibrils were observed in the dendrimer crosslinked gels. As a result, the optical transparency of the dendrimer crosslinked collagen was significantly better than that of the collagen thermal gels. The EDC and glutaraldehyde crosslinked gels were generally less transparent than those crosslinked with the dendrimers. Glucose permeation results demonstrated that the dendrimer crosslinked collagen had higher glucose permeability than natural human cornea. Dendrimer crosslinked collagen gels supported human corneal epithelial cell growth and adhesion, with no cell toxicity. In comparison, some potentially cytotoxic effects were observed with glutaraldehyde crosslinked collagen. Overall, the dendrimer crosslinked collagen gels showed promising properties that suggest that these might be suitable scaffolds for corneal tissue engineering and potentially other tissue engineering applications.