Expression and subcellular localization of efflux transporter ABCG2/BCRP in important tissue barriers of lactating dairy cows, sheep and goats.
Expression of efflux transporter ABCG2/BCRP in tissues barriers has shown to be associated with altered pharmaco- and toxicokinetics of xenobiotics. Until now, little is known about the functional expression of this transporter in dairy animals. We therefore systematically examined the expression and subcellular localization of ABCG2/BCRP in small intestine, colon, lung, liver, kidney and mammary gland in lactating cows, sheep and goats. Carrier expression was investigated by RT-PCR and Western blot analysis showing highest expression of ABCG2/BCRP in small intestine and mammary gland, high levels in liver and moderate amounts of protein in lung, colon and kidney. Regarding subcellular localization, BCRP was predominantly found at the apical plasma membrane of small intestine, colon, bronchial epithelium, bile ducts and overall in endothelial structures in all tested species. In the mammary gland, there was strong apical staining of the alveolar epithelial cells and most of the ducts in all dairy ruminants. We also detected significantly elevated protein expression in lactating mammary gland compared with nonlactating cows, sheep and goats. Our results contribute to the role of BCRP in cytoprotection and disposition in important tissue barriers and may have important implications for veterinary pharmacotherapy of dairy animals using drugs identified as BCRP substrates.


Assessing the Relative Effectiveness of Combining Self-Care with Practitioner-Delivered Complementary and Integrative Health Therapies to Improve Pain in a Pragmatic Trial.
Many health care systems are beginning to encourage patients to use complementary and integrative health (CIH) therapies for pain management. Many clinicians have anecdotally reported that patients combining self-care CIH therapies with practitioner-delivered therapies report larger health improvements than do patients using practitioner-delivered or self-care CIH therapies alone. However, we are unaware of any trials in this area. The APPROACH Study (Assessing Pain, Patient-Reported Outcomes and Complementary and Integrative Health) assesses the value of veterans participating in practitioner-delivered CIH therapies alone or self-care CIH therapies alone compared with the combination of self-care and practitioner-delivered care. The study is being conducted in 18 Veterans Health Administration sites that received funding as part of the Comprehensive Addiction and Recovery Act to expand availability of CIH therapies. Practitioner-delivered therapies under study include chiropractic care, acupuncture, and therapeutic massage, and self-care therapies include tai chi/qi gong, yoga, and meditation. The primary outcome will be improvement on the Brief Pain Inventory 6 months after initiation of CIH as compared with baseline scores. Patients will enter treatment groups on the basis of the care they receive because randomizing patients to specific CIH therapies would require withholding therapies routinely offered at VA. We will address selection bias and confounding by using sites' variations in business practices and other encouragements to receive different types of CIH therapies as a surrogate for direct randomization by using instrumental variable econometrics methods. Real-world evidence about the value of combining self-care and practitioner-delivered CIH therapies from this pragmatic trial will help guide the VA and other health care systems in offering specific nonpharmacological approaches to manage patients' chronic pain.


Emergence of microbial diversity due to cross-feeding interactions in a spatial model of gut microbial metabolism.
The human gut contains approximately 1014 bacteria, belonging to hundreds of different species. Together, these microbial species form a complex food web that can break down nutrient sources that our own digestive enzymes cannot handle, including complex polysaccharides, producing short chain fatty acids and additional metabolites, e.g., vitamin K. Microbial diversity is important for colonic health: Changes in the composition of the microbiota have been associated with inflammatory bowel disease, diabetes, obesity and Crohn's disease, and make the microbiota more vulnerable to infestation by harmful species, e.g., Clostridium difficile. To get a grip on the controlling factors of microbial diversity in the gut, we here propose a multi-scale, spatiotemporal dynamic flux-balance analysis model to study the emergence of metabolic diversity in a spatial gut-like, tubular environment. The model features genome-scale metabolic models (GEM) of microbial populations, resource sharing via extracellular metabolites, and spatial population dynamics and evolution. In this model, cross-feeding interactions emerge readily, despite the species' ability to metabolize sugars autonomously. Interestingly, the community requires cross-feeding for producing a realistic set of short-chain fatty acids from an input of glucose, If we let the composition of the microbial subpopulations change during invasion of adjacent space, a complex and stratified microbiota evolves, with subspecies specializing on cross-feeding interactions via a mechanism of compensated trait loss. The microbial diversity and stratification collapse if the flux through the gut is enhanced to mimic diarrhea. In conclusion, this in silico model is a helpful tool in systems biology to predict and explain the controlling factors of microbial diversity in the gut. It can be extended to include, e.g., complex nutrient sources, and host-microbiota interactions via the intestinal wall.


Traumatic Head Injury and the Diagnosis of Abuse: A Cluster Analysis.
Data guiding abusive head trauma (AHT) diagnosis rest on case-control studies that have been criticized for circularity. We wished to sort children with neurologic injury using mathematical algorithms, without reference to physicians' diagnoses or predetermined diagnostic criteria, and to compare the results to existing AHT data, physicians' diagnoses, and a proposed triad of findings. Unsupervised cluster analysis of an existing data set regarding 500 young patients with acute head injury hospitalized for intensive care. Three cluster algorithms were used to sort (partition) patients into subpopulations (clusters) on the basis of 32 reliable (κ > 0.6) clinical and radiologic variables. P values and odds ratios (ORs) identified variables most predictive of partitioning. The full cohort partitioned into 2 clusters. Variables substantially (P < .001 and OR > 10 in all 3 cluster algorithms) more prevalent in cluster 1 were imaging indications of brain hypoxemia, ischemia, and/or swelling; acute encephalopathy, particularly when lasting >24 hours; respiratory compromise; subdural hemorrhage or fluid collection; and ophthalmologist-confirmed retinoschisis. Variables substantially (P < .001 and OR < 0.10 in any cluster algorithm) more prevalent in cluster 2 were linear parietal skull fracture and epidural hematoma. Postpartitioning analysis revealed that cluster 1 had a high prevalence of physician-diagnosed abuse. Three cluster algorithms partitioned the population into 2 clusters without reference to predetermined diagnostic criteria or clinical opinion about the nature of AHT. Clinical difference between clusters replicated differences previously described in comparisons of AHT with non-AHT. Algorithmic partition was predictive of physician diagnosis and of the triad of findings heavily discussed in AHT literature.


Osteoinductive and Osteogenic Capacity of Freeze-Dried Bovine Bone Compared to Deproteinized Bovine Bone Mineral Scaffold in Human Umbilical Cord Mesenchymal Stem Cell Culture: An In Vitro Study.
Freeze-dried bovine bone scaffold (FDBB) or decellularized FDBB (dc-FDBB) was developed as an ideal scaffold with osteoinductive properties. This research aims to compare the osteoinductive properties marked by the expression of runt-related transcription factor-2 (RUNX2) and Osterix (OSX) and the osteogenic capacity of these scaffolds imbued with human umbilical cord mesenchymal stem cells (hUCMSCs). This study was performed in five experimental groups: a negative control group (C-) of hUCMSCs with a normal growth medium, a positive control group (C + ) of hUCMSCs with an osteogenic medium, experimental group 1 (E1) with an FDBB conditioned medium (CM), and experimental group 2 (E2) with a dc-FDBB-CM, and a third experimental group (E3) consisting of a DBBM-CM. Alizarin red staining was performed to qualitatively assess osteoinductive capacity. RUNX2 and OSX expression was quantified using real-time quantification polymerase chain reaction with two replications on day six (D6) and day 12 (D12) as fold changes. This experiment revealed that hUCMSCs were positively expressed by CD73, CD90, and CD105 but were not expressed by CD34. Alizarin red staining showed that E1 had the most calcium deposition on D6 and D12, followed by E3 and then E2 The RUNX2 and OSX expression was higher in E1 but this difference was not significant. The OSX expression in E1,E2,E3 was lower on D12 and C+ of OSX had the highest expression. There was a significant difference of fold change measured between all groups (p < 0.05), and there was no significant difference between any of the groups treated with OSX and RUNX2 on D6 and D12. FDBB osteoinduction and osteogenic capacity were higher when compared with DBBM and dc-FDBB.


Sustaining dry surfaces under water.
Rough surfaces immersed under water remain practically dry if the liquid-solid contact is on roughness peaks, while the roughness valleys are filled with gas. Mechanisms that prevent water from invading the valleys are well studied. However, to remain practically dry under water, additional mechanisms need consideration. This is because trapped gas (e.g. air) in the roughness valleys can dissolve into the water pool, leading to invasion. Additionally, water vapor can also occupy the roughness valleys of immersed surfaces. If water vapor condenses, that too leads to invasion. These effects have not been investigated, and are critically important to maintain surfaces dry under water. In this work, we identify the critical roughness scale, below which it is possible to sustain the vapor phase of water and/or trapped gases in roughness valleys - thus keeping the immersed surface dry. Theoretical predictions are consistent with molecular dynamics simulations and experiments.


Shared decision-making about cardiovascular disease medication in older people: a qualitative study of patient experiences in general practice.
To explore older people's perspectives and experiences with shared decision-making (SDM) about medication for cardiovascular disease (CVD) prevention. Semi-structured interviews with 30 general practice patients aged 75 years and older in New South Wales, Australia, who had elevated CVD risk factors (blood pressure, cholesterol) or had received CVD-related lifestyle advice. Data were analysed by multiple researchers using Framework analysis. Twenty eight participants out of 30 were on CVD prevention medication, half with established CVD. We outlined patient experiences using the four steps of the SDM process, identifying key barriers and challenges: Step 1. Choice awareness: taking medication for CVD prevention was generally not recognised as a decision requiring patient input; Step 2. Discuss benefits/harms options: CVD prevention poorly understood with emphasis on benefits; Step 3. Explore preferences: goals, values and preferences (eg, length of life vs quality of life, reducing disease burden vs risk reduction) varied widely but generally not discussed with the general practitioner; Step 4. Making the decision: overall preference for directive approach, but some patients wanted more active involvement. Themes were similar across primary and secondary CVD prevention, different levels of self-reported health and people on and off medication. Results demonstrate how older participants vary widely in their health goals and preferences for treatment outcomes, suggesting that CVD prevention decisions are preference sensitive. Combined with the fact that the vast majority of participants were taking medications, and few understood the aims and potential benefits and harms of CVD prevention, it seems that older patients are not always making an informed decision. Our findings highlight potentially modifiable barriers to greater participation of older people in SDM about CVD prevention medication and prevention in general.


In vitro models of exosome biology and toxicology: New frontiers in biomedical research.
Exosomes are secreted membrane-bound vesicles containing a cargo of curated nucleic acids, proteins, and lipids that can alter gene expression in recipient cells. Toxic agents can alter exosome synthesis and bioactive cargo composition, thus allowing exosomes to serve as biomarkers of exposure and response. While human and animal studies have identified exosome biomarkers of organ toxicity, in vitro models are ideal to examine biological mechanisms of exosome function. Here, we discuss the importance of exosomes in toxicology research and describe applications of in vitro models in advancing our understanding of their role in exposure-associated disease. This discussion of new research frontiers is in commemoration of the invaluable contributions of Dr. Daniel Acosta to the field of in vitro biology and toxicology. Emerging studies have implicated exosomes as mediators of neurodegeneration by shuttling pollutant-induced pathogenic proteins and miRNAs from afflicted neurons to neighboring cells. Exosomes also provide a mechanistic link between inhalation exposures and airway inflammation, remodeling, and systemic effects. Exosomes provide the means for toxic agents to initiate oncogenic transformation and create favorable tumor microenvironments. Furthermore, exosome-mediated drug delivery can alter drug pharmacologic profiles. Expansion in this field using in vitro models is essential to unlock the potential applications of exosome biology in toxicology.


Protein polymer hydrogels: effects of endotoxin on biocompatibility.
Protein polymer-based hydrogels have shown potential for tissue engineering applications, but require biocompatibility testing for in vivo use. Enzymatically crosslinked protein polymer-based hydrogels were tested in vitro and in vivo to evaluate their biocompatibility. Endotoxins present in the hydrogel were removed by Trition X-114 phase separation. The reduction of endotoxins decreased TNF-α production by a macrophage cell line in vitro; however, significant inflammatory response was still present compared to collagen control gels. A branched PEG molecule and dexamethasone were added to the hydrogel to reduce the response. In vitro testing showed a decrease in the TNF-α levels with the addition of dexamethasone. In vivo implantations into the epididymal fat pad of C57/BL6 mice, however, indicated a decreased inflammatory mediated immune response with a hydrogel treated with both PEGylation and endotoxin reduction. This study demonstrates the importance of endotoxin testing and removal in determining the biocompatibility of biomaterials.


Extracellular vesicles as therapeutic tools in cardiovascular diseases.
Extracellular vesicles (EVs), including microvesicles (MVs) and exosomes, are small vesicles secreted from a wide variety of cells. Whereas MVs are particles released by the outward budding of the plasma membrane, exosomes are derived from endocytic compartments. Secretion of EVs can be enhanced by specific stimuli, and increased plasma circulating levels of EVs have been correlated with pathophysiological situations. MVs, already present in the blood of healthy individuals, are considerably elevated in several cardiovascular diseases associated with inflammation, suggesting that they can mediate deleterious effects such as endothelial dysfunction or thrombosis. Nonetheless, very recent studies also demonstrate that MVs may act as biological information vectors transferring proteins or genetic material to maintain cell homeostasis, favor cell repair, or even promote angiogenesis. Additionally, exosomes have also been shown to have pro-angiogenic and cardio-protective properties. These beneficial effects, therefore, reveal the potential therapeutical use of EVs in the field of cardiovascular medicine and regenerative therapy. In this review, we will provide an update of cellular processes modulated by EVs of specific interest in the treatment of cardiovascular pathologies. A special focus will be made on the morphogen sonic hedgehog (Shh) associated with EVs (EVs(Shh+)), which have been shown to mediate many pro-angiogenic effects. In addition to offer a potential source of cardiovascular markers, therapeutical potential of EVs reveal exciting opportunities to deliver specific agents by non-immunogenic means to cardiovascular system.


TBK1 gene duplication and normal-tension glaucoma.
Normal-tension glaucoma (NTG) is a common cause of vision loss. To investigate the role of TANK binding kinase 1 (TBK1) gene duplications in NTG to gain insights into the causes of glaucoma that occurs at low intraocular pressure (IOP). In this multicenter case-control study, we investigated patients who met the criteria for NTG, including glaucomatous optic neuropathy, visual field defects, and maximum recorded untreated IOP of 21 mm Hg or less, and matched controls. Participants (N = 755) were recruited from Southampton, United Kingdom (180 patients and 178 controls), Rochester, Minnesota (65 patients and 12 controls), New York, New York (96 patients and 16 controls), and Iowa City, Iowa (208 controls). Detection of TBK1 gene duplications and comparison of the extent of the identified DNA that is duplicated with prior TBK1 copy number variations associated with NTG. A TBK1 gene duplication was detected in 1 of 96 patients (1.0%) from New York and none of the controls. Analysis of duplication borders with comparative genome hybridization demonstrated that this patient has a novel duplication that has not been previously reported. No gene duplications were detected in any of the other cohorts of patients or controls. Duplication of the TBK1 gene is a rare cause of NTG. The identification of another case of NTG attributed to TBK1 gene duplication strengthens the case that this mutation causes glaucoma.


Widely Used Herpes Simplex Virus 1 ICP0 Deletion Mutant Strain dl1403 and Its Derivative Viruses Do Not Express Glycoprotein C Due to a Secondary Mutation in the gC Gene.
Herpes simplex virus 1 (HSV-1) ICP0 is a multi-functional phosphoprotein expressed with immediate early kinetics. An ICP0 deletion mutant, HSV-1 dl1403, has been widely used to study the roles of ICP0 in the HSV-1 replication cycle including gene expression, latency, entry and assembly. We show that HSV-1 dl1403 virions lack detectable levels of envelope protein gC, and that gC is not synthesized in infected cells. Sequencing of the gC gene from HSV-1 dl1403 revealed a single amino acid deletion that results in a frameshift mutation. The HSV-1 dl1403 gC gene is predicted to encode a polypeptide consisting of the original 62 N-terminal amino acids of the gC protein followed by 112 irrelevant, non-gC residues. The mutation was also present in a rescuant virus and in two dl1403-derived viruses, D8 and FXE, but absent from the parental 17+, suggesting that the mutation was introduced during the construction of the dl1403 virus, and not as a result of passage in culture.


Bimodal distribution of renal cytochrome P450 3A activity in humans.
It has been proposed that excessive intrarenal conversion of cortisol to 6 beta-hydroxycortisol by CYP3A may mediate increased tubular reabsorption of sodium, leading to a state of mild volume expansion and the clinical phenotype of salt-sensitive hypertension. Therefore, we characterized CYP3A activity in a bank of microsomes from human kidneys using the formation of 1'-hydroxymidazolam (1'-OHM) as a prototypical CYP3A-catalyzed reaction. Maximal rates of metabolite formation occurred at midazolam concentrations of 12.5-50 microM; higher concentrations resulted in dramatic substrate inhibition. At 12.5 microM midazolam, 4 of 27 kidneys exhibited relatively high mean +/- standard deviation 1'-OHM formation rate (184.0 +/- 14.4 pmol/hr/mg) compared with the remaining 23 samples, which had a mean formation rate of (10.1 +/- 6.4 pmol/hr/mg). Triacetyloleandomycin and anti-CYP3A antibody inhibited midazolam hydroxylation by 53% and 57%, respectively. The correlation between CYP3A5 content, determined through immunoblotting, and 1'-OHM formation rate was high (r2 = 0.84, 24 experiments). The expressions of mRNA corresponding to CYP3A3, CYP3A4, CYP3A5, and CYP3A7 were determined through polymerase chain reaction with specific oligonucleotides as primers. All kidneys examined (25 experiments) expressed CYP3A5 protein and contained the corresponding CYP3A5 mRNA. CYP3A4 mRNA was detected in 40% of the kidney samples, and 70% of those that contained detectable CYP3A4 mRNA also expressed detectable levels of the corresponding protein. Therefore, in contrast to hepatic tissue, in which CYP3A4 is universally expressed, CYP3A5 is the ubiquitously expressed member of the CYP3A family in renal tissue. The distribution of enzyme activity and protein content suggests bimodality and may represent induction of CYP3A5 in a select population and/or a genetically determined organ-specific pattern of expression.


Associations between circulating proteins and corresponding genes expressed in coronary thrombi in patients with acute myocardial infarction.
Several genes are expressed in aspirated coronary thrombi in acute myocardial infarction (AMI), exhibiting dynamic changes along ischemic time. Whether soluble biomarkers reflect the local gene environment and ischemic time is unclear. We explored whether circulating biomarkers were associated with corresponding coronary thrombi genes and total ischemic time. In 33 AMI patients undergoing percutaneous coronary intervention (PCI), blood samples were collected within 6-24h for markers related to plaque rupture (metalloproteinase 9, tissue inhibitor of metalloproteinases 1), platelet and endothelial cell activation (P-selectin, CD40 ligand, PAR-1), hemostasis (tissue factor, tissue plasminogen activator, plasminogen activator inhibitor 1, free and total tissue factor pathway inhibitor, D-dimer, prothrombin fragment 1+2), inflammation (interleukin 8 and 18, fractalkine, monocyte chemoattractant protein 1 (MCP-1), CXCL1, pentraxin 3, myeloperoxidase) and galectin 3, caspase 8 and epidermal growth factor (EGF). Laboratory analyses were performed by Proximity Extension Assay (Proseek Multiplex CVD I(96 × 96)), ELISAs and RT-PCR. Only circulating P-selectin correlated to the corresponding P-selectin gene expression in thrombi (r=0.530, p=0.002). Plasma galectin 3, fractalkine, MCP-1 and caspase 8 correlated inversely to ischemic time (r=-0.38-0.50, all p <0.05), while plasma MCP-1, galectin 3 and EGF were higher at short (≤ 4 h) vs. long (>4h) ischemic time (all p <0.05). The dynamic changes in circulating mediators along ischemic time were not reflected in the profile of locally expressed genes. These observations indicate a locally confined milieu within the site of atherothrombosis, which may be important for selective therapy.


Gender-specific influence of health behaviors on academic performance in Spanish adolescents: the AFINOS study.
New paradigms based on the multifactorial etiology of chronic diseases and behavioral outcomes suggest that a combination of health behaviors may have more impact on the outcome of interest than any single factor. To examine the independent and combined influence of four health behaviors on school performance in Spanish adolescents. A total of 1825 Spanish adolescents reported their grades in Language and Literature (LL) and Math. Body mass index, family structure and school-related factors (attitude to school, need to repeat > 1-yr and absenteeism) were self-reported. Adolescents were dichotomized as healthy or unhealthy based on meeting or not meeting lifestyle recommendations on physical activity, TV viewing, sleep and fruit intake. Each adolescent was also scored according to the number of healthy recommendations fulfilled. In boys, there were no associations between health behaviors and academic performance. Good academic performance in girls was associated with physical activity (P < 0.05) or fruit consumption (P < 0.05). Moreover, girls who scored 3-4 health behaviors showed higher odds of passing LL (OR = 3.18, P < 0.001), Math (OR = 1.75, P = 0.028) or LL+Math (OR = 2.32, P = 0.001) compared with those with 0-1 health behaviors. All the analyses were adjusted by weight status, family context and different school-related factors. A combination of health behaviors may have a positive influence on academic performance in adolescent girls.


New Insights into the Impact of Genome-Wide Copy Number Variations on Complex Congenital Heart Disease in Saudi Arabia.
Congenital heart diseases (CHDs) are complex traits that manifest in diverse clinical phenotypes such as the Tetralogy of Fallot (TOF), valvular and ventricular/atrial septal defects. Genetic mechanisms of CHDs have remained largely unclear to date. Copy number variations (CNVs) have been implicated in many complex diseases but their impact has not been examined extensively in various forms of CHD lesions. We report in this study, to the best of our knowledge, the largest cohort of Saudi Arab CHD patients to date who were evaluated using genome-wide CNV analysis. In a sample of 134 Saudi Arab patients with CHD, 66 exhibited pathogenic or likely pathogenic CNVs. Notably, 21 copy number gains and 11 copy number losses were detected that encompassed 141 genes and 146 genes, respectively. The most frequent gains were on 17q21.31, 8p11.21, and 22q11.23, whereas the losses were primarily localized to 16p11.2. Interestingly, all lesions have had gains at 17q21.31. Septal defects had also gains at 8p11.21 and 22q11.23, valvular lesions at 8p11.21, 22q11.23, and 2q13, and TOF at 16p11.2. Functional and network analyses demonstrated that cardiovascular and nervous system development and function as well as cell death/survival were most significantly associated with CNVs, thus highlighting the potentially important genes likely to be involved in CHD, including NPHP1, PLCB1, KANSL1, and NR3C1. In conclusion, this genome-wide analysis identifies a high frequency of CNVs mostly in patients with septal defects, primarily influencing cardiovascular developmental and functional pathways, thereby offering a deeper insight into the complex networks involved in CHD pathogenesis.


A dual role of lipocalin 2 in the apoptosis and deramification of activated microglia.
Activated microglia are thought to undergo apoptosis as a self-regulatory mechanism. To better understand molecular mechanisms of the microglial apoptosis, apoptosis-resistant variants of microglial cells were selected and characterized. The expression of lipocalin 2 (lcn2) was significantly down-regulated in the microglial cells that were resistant to NO-induced apoptosis. lcn2 expression was increased by inflammatory stimuli in microglia. The stable expression of lcn2 as well as the addition of rLCN2 protein augmented the sensitivity of microglia to the NO-induced apoptosis, while knockdown of lcn2 expression using short hairpin RNA attenuated the cell death. Microglial cells with increased lcn2 expression were more sensitive to other cytotoxic agents as well. Thus, inflammatory activation of microglia may lead to up-regulation of lcn2 expression, which sensitizes microglia to the self-regulatory apoptosis. Additionally, the stable expression of lcn2 in BV-2 microglia cells induced a morphological change of the cells into the round shape with a loss of processes. Treatment of primary microglia cultures with the rLCN2 protein also induced the deramification of microglia. The deramification of microglia was closely related with the apoptosis-prone phenotype, because other deramification-inducing agents such as cAMP-elevating agent forskolin, ATP, and calcium ionophore also rendered microglia more sensitive to cell death. Taken together, our results suggest that activated microglia may secrete LCN2 protein, which act in an autocrine manner to sensitize microglia to the self-regulatory apoptosis and to endow microglia with an amoeboid form, a canonical morphology of activated microglia in vivo.


Potential Risk Factors for the Development of Self-Injurious Behavior among Infants at Risk for Autism Spectrum Disorder.
Prevalence of self-injurious behavior (SIB) is as high as 50% among children with autism spectrum disorder (ASD). Identification of risk factors for the development of SIB is critical to early intervention and prevention. However, there is little empirical research utilizing a prospective design to identify early risk factors for SIB. The purpose of this study was to evaluate behavioral characteristics predicting SIB at age 2 years among 235 infants at high familial risk for ASD. Logistic regression results indicated that presence of SIB or proto-SIB and lower developmental functioning at age 12 months significantly predicted SIB at 24 months. A pattern of persistent SIB over this period was associated with a diagnosis of autism and poorer cognitive and adaptive outcomes.


Enlargement of Lyt-2-positive T cells is associated with functional impairment and autoimmune hemolytic anemia in New Zealand Black mice.
We investigated the relationship between the increased cell diameter of Lyt-2+ T cells and the development of autoimmune disease in aging NZB and NZB X NZW F1 hybrid (BW) mice. Individual animals were analyzed for Lyt-2+ T cell size (by narrow-angle forward light scatter), anti-erythrocyte autoantibodies, anemia, proteinuria, and splenomegaly. The peak light scatter of the Lyt-2+ T cells correlated with the level of anti-erythrocyte autoantibodies and severity of hemolytic anemia, but not with proteinuria or splenomegaly. The cell size of this T cell subset did not increase in old BW or in NZB mice homozygous for the xid gene (NZB.xid). The in vivo administration of bacterial lipopolysaccharide to young NZB mice did not stimulate the enlargement of Lyt-2+ T cells. Ly-2+ T cells from old NZB mice could be stimulated by concanavalin A (Con A) to express interleukin 2 (IL 2) receptors and to synthesize DNA in vitro. However, in vivo administration of Con A to old NZB mice did not induce the expression of IL 2 receptors on Lyt-2+ T cells. Further, in vivo T suppressor function was impaired in old NZB mice with enlarged Lyt-2+ T cells. Thus, the enlargement of Lyt-2+ T cells in old NZB mice appears related to impaired T cell function in vivo and is associated with the development of anti-erythrocyte autoantibodies and autoimmune hemolytic anemia.


Work-related asthma in teachers in Connecticut: association with chronic water damage and fungal growth in schools.
To review a series of 55 teachers from schools in Connecticut who presented consecutively to our clinic, and to summarize our clinical experience with work-related disease in this population, which could lead to a more formal study. Retrospective chart review; data were abstracted concerning clinical and environmental factors and entered into a statistical spreadsheet program (JMP for Windows). Of the 55 educators, 22 were diagnosed with upper respiratory syndromes (rhinitis or sinusitis), three with bronchitis, and 23 with asthma. Of the 23 with asthma, 20 presented with active or symptomatic asthma, and seven of these were incident cases of occupational asthma. In addition, four cases (7%) of granulomatous lung disease (two hypersensitivity pneumonitis and two sarcoidosis) were diagnosed. Finally, three patients (5%) received only nonrespiratory diagnoses (panic disorder, sicca syndrome, and vertigo). In 33 work-places, the exposures of concern were predominantly related to ongoing "dampness" or visible mold growth. The remaining 22 work-places were "dry." Symptoms varied according to the work-place environment, with more patients from water damaged (vs dry) work-places having upper respiratory symptoms (76% vs 45%) and asthma (45% vs 23%). All seven patients with incident asthma and all four patients with interstitial lung disease worked in schools with documented water incursion. Work-place exposures in water damaged school buildings are risk factors for development of work-related lower respiratory disease in schoolteachers and staff. Identification of such high-risk environments can be done by a simple but thorough qualitative evaluation during a walk-through inspection, and it should not require air sampling or surface sampling protocols for microbial contaminants.


The chronic effects of cannabis on memory in humans: a review.
Memory problems are frequently associated with cannabis use, in both the short- and long-term. To date, reviews on the long-term cognitive sequelae of cannabis use have examined a broad range of cognitive functions, with none specifically focused on memory. Consequently, this review sought to examine the literature specific to memory function in cannabis users in the nontoxicated state with the aim of identifying the existence and nature of memory impairment in cannabis users and appraising potentially related mediators or moderators. Literature searches were conducted to extract well-controlled studies that investigated memory function in cannabis users outside of the acute intoxication period, with a focus on reviewing studies published within the past 10 years. Most recent studies have examined working memory and verbal episodic memory and cumulatively, the evidence suggests impaired encoding, storage, manipulation and retrieval mechanisms in long-term or heavy cannabis users. These impairments are not dissimilar to those associated with acute intoxication and have been related to the duration, frequency, dose and age of onset of cannabis use. We consider the impact of not only specific parameters of cannabis use in the manifestation of memory dysfunction, but also such factors as age, neurodevelopmental stage, IQ, gender, various vulnerabilities and other substance-use interactions, in the context of neural efficiency and compensatory mechanisms. The precise nature of memory deficits in cannabis users, their neural substrates and manifestation requires much further exploration through a variety of behavioural, functional brain imaging, prospective and genetic studies.


Preconception care: caffeine, smoking, alcohol, drugs and other environmental chemical/radiation exposure.
As providing health education, optimizing nutrition, and managing risk factors can be effective for ensuring a healthy outcome for women and her yet un-conceived baby, external influences play a significant role as well. Alcohol, smoking, caffeine use and other similar lifestyle factors, have now become an integral part of the daily life of most men and women, who use/misuse one or more of these harmful substances regularly despite knowledge of their detrimental effects. The adverse health outcomes of these voluntary and involuntary exposures are of even greater concern in women of child bearing age where the exposure has the potential of inflicting harm to two generations. This paper is examining the available literature for the possible effects of caffeine consumption, smoking, alcohol or exposure to chemicals may have on the maternal, newborn and child health (MNCH). A systematic review and meta-analysis of the evidence was conducted to ascertain the possible impact of preconception usage of caffeine, tobacco, alcohol and other illicit drugs; and exposure to environmental chemicals and radiant on MNCH outcomes. A comprehensive strategy was used to search electronic reference libraries, and both observational and clinical controlled trials were included. Cross-referencing and a separate search strategy for each preconception risk and intervention ensured wider study capture. Heavy maternal preconception caffeine intake of >300 mg/d significantly increase the risk of a subsequent fetal loss by 31% (95% CI: 8-58%). On the other hand, preconception alcohol consumption leads to non-significant 30% increase in spontaneous abortion (RR 1.30; 95% CI: 0.85-1.97). Preconception counselling can lead to a significant decrease in the consumption of alcohol during the first trimester (OR 1.79; 95% CI: 1.08-2.97). Periconception smoking, on the other hand, was found to be associated with an almost 3 times increased risk of congenital heart defects (OR 2.80; 95% CI 1.76-4.47). While the review found limited evidence of preconception environmental exposure on maternal, newborn and child health outcomes, occupational exposure in female radiation workers before conception showed an increased impact in risk of early miscarriages. Identification of substance abuse and environmental history during preconception period provides an opportunity to assist women in reducing major health risks and identify key determinants of healthy pregnancy. Studies have shown that the aversion and prevention of exposure feasibility can play an important role in improving the health of women and their families, however, the results should be interpreted with great caution as there were few studies in each section. Therefore, there is a need for more rigorous studies to test the hypotheses.


Development, Evaluation, and Sensitivity Analysis of Parametric Finite Element Whole-Body Human Models in Side Impacts.
Occupant stature and body shape may have significant effects on injury risks in motor vehicle crashes, but the current finite element (FE) human body models (HBMs) only represent occupants with a few sizes and shapes. Our recent studies have demonstrated that, by using a mesh morphing method, parametric FE HBMs can be rapidly developed for representing a diverse population. However, the biofidelity of those models across a wide range of human attributes has not been established. Therefore, the objectives of this study are 1) to evaluate the accuracy of HBMs considering subject-specific geometry information, and 2) to apply the parametric HBMs in a sensitivity analysis for identifying the specific parameters affecting body responses in side impact conditions. Four side-impact tests with two male post-mortem human subjects (PMHSs) were selected to evaluate the accuracy of the geometry and impact responses of the morphed HBMs. For each PMHS test, three HBMs were simulated to compare with the test results: the original Total Human Model for Safety (THUMS) v4.01 (O-THUMS), a parametric THUMS (P-THUMS), and a subject-specific THUMS (S-THUMS). The P-THUMS geometry was predicted from only age, sex, stature, and BMI using our statistical geometry models of skeleton and body shape, while the S-THUMS geometry was based on each PMHS's CT data. The simulation results showed a preliminary trend that the correlations between the PTHUMS- predicted impact responses and the four PMHS tests (mean-CORA: 0.84, 0.78, 0.69, 0.70) were better than those between the O-THUMS and the normalized PMHS responses (mean-CORA: 0.74, 0.72, 0.55, 0.63), while they are similar to the correlations between S-THUMS and the PMHS tests (mean-CORA: 0.85, 0.85, 0.67, 0.72). The sensitivity analysis using the PTHUMS showed that, in side impact conditions, the HBM skeleton and body shape geometries as well as the body posture were more important in modeling the occupant impact responses than the bone and soft tissue material properties and the padding stiffness with the given parameter ranges. More investigations are needed to further support these findings.


Alterations in preload and ejection phase indices of left ventricular performance,.
Ejection phase indices such as mean velocity of circumferential fiber shortening (V-cf), ejection fraction (EF), and mean normalized systolic ejection rate (MSER) are thought to reflect the level of left ventricular (LV) inotropic state. Although known to be influenced by changes in afterload, their response to altered preload is less certain. This question was examined in ten normal subjects studied in the supine position and after 10 min of 75 degree head-up tilt. Echo VL diameter (D) and carotid pulse recordings were used to calculate LV ejection time, V-cf, MSER, LV end-diastolic and stroke volumes (EDV and SV, by D3 formula), and ejection fraction. Systemic blood pressure (BP)was measured by cuff, and heart rate (HR) counted from the ECG. Calculated LVEDV decreased with tilt (115 plus or minus 8 to 82 plus or minus 7 ml, P smaller than 0.001), as did SV (81 plus or minus 6 to 54 plus or minus 6 ml, P smaller than 0.001), and ejection time (0.31 plus or minus 0.01 to 0.27 plus or minus 0.00 sec, P smaller than 0.001), but there was no significant change in systolic or diastolic BP or HR. V-cf, EF, and MSER were not singificantly altered by tilt (1.09 plus or minsu 0.04 to 1.12 plus or minus 0.05 circ/sec; 0.70 plus or minus 0.02 to 0.65 plus or minus 0.02; and 2.29 plus or minus 0.07 to 2.43 plus or minus 0.08 end-diastolic volumes/sec, respectively). V-cf, MSER, and EF appearrelatively unaffected by acute alterations in preload. That compensatory sympathetic responses blunted potential changes cannot be excluded, although HR was not found to have increased.


Coagulation Profile in Patients with Different Etiologies for Cushing Syndrome: A Prospective Observational Study.
Previous studies reported a higher prevalence of venous-thromboembolic events among patients with Cushing disease (CD) compared to those with ACTH-independent Cushing syndrome (CS) from adrenal sources. The objective of the current study was to evaluate the coagulation profile of patients with CS from different etiologies. A prospective observational study was conducted at a clinical research center. The study included adult patients admitted for evaluation of suspected CS (n=85), that were divided into 3 groups: CD (n=22), ACTH-independent CS from an adrenal tumor/hyperplasia (adrenal CS, n=21), and a control group consisting of subjects with negative screening for CS (rule-out CS, n=42). Coagulation profiles were drawn before and 8.5±4.3 months after surgery (trans-sphenoidal or adrenalectomy, n=18), and included fibrinogen, Factor VIII (FVIII), von Willebrand factor antigen (vWF:Ag), plasminogen activator inhibitor-1 (PAI-1), antithrombin III (ATIII), Protein C (PC), Protein S (PS), α2-antiplasmin (α2AP), and aPTT measurements. Patients with CD had higher baseline mean cortisol levels, ATIII activity and vWF:Ag levels compared with adrenal CS. Differences in ATIII activity and vWF:Ag levels remained even after controlling for BMI, and ATIII after also controlling for 24-h urinary free cortisol collections. Our study showed for the first time the differences in coagulation profiles between various etiologies of CS. We assume that the higher cortisol burden among CD patients may explain the differences found in the coagulation profile as well as the higher risk for VTE compared with primary adrenal CS patients.


Component-resolved diagnosis of kiwifruit allergy with purified natural and recombinant kiwifruit allergens.
Kiwifruit is one of the most common causes of food allergic reactions. Component-resolved diagnostics may enable significantly improved detection of sensitization to kiwifruit. To evaluate the use of individual allergens for component-resolved in vitro diagnosis of kiwifruit allergy. Thirty patients with a positive double-blind placebo-controlled food challenge to kiwifruit, 10 atopic subjects with negative open provocation to kiwifruit, and 5 nonatopic subjects were enrolled in the study. Specific IgE to 7 individual allergens (nAct d 1-5 and rAct d 8-9) and allergen extracts was measured by ImmunoCAP. The diagnostic sensitivities of the commercial extract and of the sum of single allergens were 17% and 77%, respectively, whereas diagnostic specificities were 100% and 30%. A combination of the kiwi allergens Act d 1, Act d 2, Act d 4, and Act d 5 gave a diagnostic sensitivity of 40%, whereas diagnostic specificity remained high (90%). Exclusion of the Bet v 1 homolog recombinant (r) Act d 8 and profilin rAct d 9 from this allergen panel reduced sensitivity to 50% but increased specificity to 40%. Kiwifruit-monosensitized patients reacted more frequently (P < .001) with Act d 1 than polysensitized patients, whereas the latter group reacted more frequently with rAct d 8 (P = .004). Use of single kiwifruit allergen ImmunoCAP increases the quantitative test performance and diagnostic sensitivity compared with the commercial extract. Bet v 1 homolog and profilin are important allergens in pollen-related kiwifruit allergy, whereas actinidin is important in monoallergy to kiwifruit, in which symptoms are often more severe.


A spatial and temporal map of C. elegans gene expression.
The C. elegans genome has been completely sequenced, and the developmental anatomy of this model organism is described at single-cell resolution. Here we utilize strategies that exploit this precisely defined architecture to link gene expression to cell type. We obtained RNAs from specific cells and from each developmental stage using tissue-specific promoters to mark cells for isolation by FACS or for mRNA extraction by the mRNA-tagging method. We then generated gene expression profiles of more than 30 different cells and developmental stages using tiling arrays. Machine-learning-based analysis detected transcripts corresponding to established gene models and revealed novel transcriptionally active regions (TARs) in noncoding domains that comprise at least 10% of the total C. elegans genome. Our results show that about 75% of transcripts with detectable expression are differentially expressed among developmental stages and across cell types. Examination of known tissue- and cell-specific transcripts validates these data sets and suggests that newly identified TARs may exercise cell-specific functions. Additionally, we used self-organizing maps to define groups of coregulated transcripts and applied regulatory element analysis to identify known transcription factor- and miRNA-binding sites, as well as novel motifs that likely function to control subsets of these genes. By using cell-specific, whole-genome profiling strategies, we have detected a large number of novel transcripts and produced high-resolution gene expression maps that provide a basis for establishing the roles of individual genes in cellular differentiation.


Hand fine motor skill disability correlates with dementia severity.
The progression of dementia, which impairs motor skills and cognitive function, is a warning of greater disability. The present study investigated the association between hand fine motor skills, assessed according to the Functioning Disability Evaluation Scale - Adult Version (FUNDES-Adult), and dementia severity. People with mild and moderate to severe dementia were identified from the Taiwan Data Bank of Persons with Disability. The FUNDES-Adult was assessed for all enrollees, and the following hand fine motor skills were evaluated: pen-holding, buttoning, and knotting. Statistical analysis was performed using SAS, and P values < 0.05 were considered significant. Disability in all the 3 fin. motor skills was significantly greater in patients with moderate to severe dementia than in those with mild dementia. Disability in any of the skills was sensitive to distinguish mild from moderate to severe dementia (sensitivity: 78.1 %, specificity: 55.2 %, area under the curve: 0.739, 95 % confidence interval [CI]: 0.734-0.745). Those with fine motor skill disability were at a significantly higher risk of moderate to severe dementia (odds ratio: 3.71, 95 % CI: 3.53-3.90, P < .001). Hand fine motor skill disability was more prevalent in patients with moderate to severe dementia than in patients with mild dementia. A straightforward motor skill assessment can serve as a screening tool in the community to detect the progression of dementia.


Comparison of regional patch collection vs. whole body washdown for measuring sweat sodium and potassium loss during exercise.
This study compared simultaneous whole body washdown (WBW) and regional skin surface (REG) sweat collections to generate regression equations to predict WBW sweat Na(+) concentration ([Na(+)]) and K(+) concentration ([K(+)]) from single- and five-site REG sweat patch collections. Athletes (10 men, 10 women) cycled in a plastic chamber for 90 min in the heat. Before exercise, the subject and bike were washed with deionized water. After the onset of sweating, sterile patches were attached to the forearm, back, chest, forehead, and thigh and removed on saturation. After exercise, the subject and bike were washed with ammonium sulfate solution to collect all sweat electrolyte loss and determine the volume of unevaporated sweat. All individual patch sites and five-site REG (weighted for local sweat rate and body surface area) were significantly (P = 0.000) correlated with WBW sweat [Na(+)]. The equation for predicting WBW sweat [Na(+)] from five-site REG was y = 0.68x + 0.44 [r = 0.97, intraclass correlation coefficient (ICC) = 0.70] and did not differ between sexes. There were sex differences in the regression results between five-site REG and WBW sweat [K(+)] (men: y = 0.74x + 0.30, r = 0.89, ICC = 0.73; women: y = 0.04x + 3.18, r = 0.03, ICC = 0.00). Five-site REG sweat [Na(+)] and [K(+)] significantly overestimated that of WBW sweat (59 +/- 27 vs. 41 +/- 19 meq/l, P = 0.000 and 4.4 +/- 0.7 vs. 3.6 +/- 0.7 meq/l, P = 0.000, respectively). For both sexes, the best sites for predicting WBW sweat [Na(+)] and [K(+)] were the thigh (1 +/- 8 meq/l < WBW, P = 1.000, y = 0.75x + 11.37, r = 0.96, ICC = 0.93) and chest (0.2 +/- 0.3 meq/l > WBW, P = 1.000, y = 0.76x + 0.55, r = 0.89, ICC = 0.87), respectively. In conclusion, regression equations can be used to accurately and reliably predict WBW sweat [Na(+)] and [K(+)] from REG sweat collections when study conditions and techniques are similar to that of the present protocol.


Characteristics of adult primary care patients as predictors of future health services charges.
Utilization risk assessment is potentially useful for allocation of health care resources, but precise measurement is difficult. Test the hypotheses that health-related quality of life (HRQOL), severity of illness, and diagnoses at a single primary care visit are comparable case-mix predictors of future 1-year charges in all clinical settings within a large health system, and that these predictors are more accurate in combination than alone. Longitudinal observational study in which subjects' characteristics were measured at baseline, and their outpatient clinic visits and charges and their inpatient hospital days and charges were tracked for 1 year. Adult primary care patients. Duke Health Profile for HRQOL, Duke Severity of Illness Checklist for severity of illness, and Johns Hopkins Ambulatory Care Groups for diagnostic groups classification. Of 1,202 patients, 84.4% had follow up in the primary care clinic, 63.2% in subspecialty clinics, 14.8% in the emergency room, and 9.6% in the hospital. Of $6,290,775 total charges, $779,037 (12.2%) was for follow-up primary care. The highest accuracy was found for predicting primary care charges, where R2 for predictors ranged from 0.083 for medical record auditor-reported severity of illness to 0.107 for HRQOL. When predictors were combined, the highest R2 of 0.125 was found for the combination of HRQOL and diagnostic groups. Baseline HRQOL, severity of illness, and diagnoses were comparable predictors of 1-year health services charges in all clinical sites but most predictive for primary care charges, and were more accurate in combination than alone.


Evaluation of microwave oven heating for prediction of drug-excipient compatibilities and accelerated stability studies.
Microwave ovens have been used extensively in organic synthesis in order to accelerate reaction rates. Here, a set up comprising a microwave oven combined with silicon carbide (SiC) plates for the controlled microwave heating of model formulations has been applied in order to investigate, if a microwave oven is applicable for accelerated drug stability testing. Chemical interactions were investigated in three selected model formulations of drug and excipients regarding the formation of ester and amide reaction products. In the accelerated stability studies, a design of experiments (DoE) approach was applied in order to be able to rank excipients regarding reactivity: Study A: cetirizine with PEG 400, sorbitol, glycerol and propylene glycol. Study B: 6-aminocaproic acid with citrate, acetate, tartrate and gluconate. Study C: atenolol with citric, tartaric, malic, glutaric, and sorbic acid. The model formulations were representative for oral solutions (co-solvents), parenteral solutions (buffer species) and solid dosage forms (organic acids applicable for solubility enhancement). The DoE studies showed overall that the same impurities were generated by microwave oven heating leading to temperatures between 150°C and 180°C as compared to accelerated stability studies performed at 40°C and 80°C using a conventional oven. Ranking of the reactivity of the excipients could be made in the DoE studies performed at 150-180°C, which was representative for the ranking obtained after storage at 40°C and 80°C. It was possible to reduce the time needed for drug-excipient compatibility testing of the three model formulations from weeks to less than an hour in the three case studies. The microwave oven is therefore considered to be an interesting alternative to conventional thermal techniques for the investigation of drug-excipient interactions during preformulation.


Systems biology identifies preserved integrity but impaired metabolism of mitochondria due to a glycolytic defect in Alzheimer's disease neurons.
Mitochondrial dysfunction is implicated in most neurodegenerative diseases, including Alzheimer's disease (AD). We here combined experimental and computational approaches to investigate mitochondrial health and bioenergetic function in neurons from a double transgenic animal model of AD (PS2APP/B6.152H). Experiments in primary cortical neurons demonstrated that AD neurons had reduced mitochondrial respiratory capacity. Interestingly, the computational model predicted that this mitochondrial bioenergetic phenotype could not be explained by any defect in the mitochondrial respiratory chain (RC), but could be closely resembled by a simulated impairment in the mitochondrial NADH flux. Further computational analysis predicted that such an impairment would reduce levels of mitochondrial NADH, both in the resting state and following pharmacological manipulation of the RC. To validate these predictions, we utilized fluorescence lifetime imaging microscopy (FLIM) and autofluorescence imaging and confirmed that transgenic AD neurons had reduced mitochondrial NAD(P)H levels at rest, and impaired power of mitochondrial NAD(P)H production. Of note, FLIM measurements also highlighted reduced cytosolic NAD(P)H in these cells, and extracellular acidification experiments showed an impaired glycolytic flux. The impaired glycolytic flux was identified to be responsible for the observed mitochondrial hypometabolism, since bypassing glycolysis with pyruvate restored mitochondrial health. This study highlights the benefits of a systems biology approach when investigating complex, nonintuitive molecular processes such as mitochondrial bioenergetics, and indicates that primary cortical neurons from a transgenic AD model have reduced glycolytic flux, leading to reduced cytosolic and mitochondrial NAD(P)H and reduced mitochondrial respiratory capacity.


Determination and comparison of mean random urine calcium between children with vesicoureteral reflux and those with improved vesicoureteral reflux.
Urinary reflux can cause irreversible complications such as reflux nephropathy and ESRD. Diagnostic imaging such as voiding cystourethrogram (VCUG) is invasive and causes irradiation. Several studies have shown that markers such as urine IL-8 or serum Procalcitonin might be useful for the diagnosis of vesico-ureteral reflux (VUR) as a substitution for invasive methods. The aim of this study was to determine and compare the mean urine Ca/Cr ratio and hypercalciuria between two groups of children aged 2-10 years affected by VUR. This is a cross-sectional study in which 32 children having the following entrance criteria were included: children 2-10 years old not affected by urinary tract infection during the last three months and their reflux or recovery having been diagnosed by VCUG or direct radionuclide cystography, divided into two groups of affected by reflux and recovered from reflux. Then, the point urine specimens were collected with permission of their parents in a single laboratory and urine Ca and Cr for each specimen were measured. The Ca/Cr ratio was calculated for each child to evaluate Ca excretion from the urine, which was possible without collecting the 24-hours urine. The Ca/Cr ratio mean and the hypercalciuria were compared between the groups. To analyze our data, the Mann-Whiney test and the Chi-square test were used, using SPSS V.15. Thirty-two children, including 18 children recovered from reflux and 14 affected by reflux, were entered in our study. The Ca/Cr ratio mean was 0.692 ± 1.874 for the affected and 0.118 ± 0.187 for the recovered group. The Ca/Cr ratio mean for the affected male and female groups was 0.012 ± 0.008 and 0.805 ± 1.0913, respectively. This amount was 0.0175 ± 0.01767 for the recovered male and 0.131 ± 0.195 for the recovered female group. There was no significant statistical difference between the groups with respect to the Ca/Cr ratio mean, but there was a significant statistical difference between the two groups of females (P-value = 0.026). Also, there was no significant statistical difference between the two groups of males relative frequency of hypercalciuria was significantly higher in the group affected by reflux than in the recovered from reflux group (P-value = 0.017). Urine Ca excretion is elevated in children with reflux, and may be more useful as an appropriate marker for the diagnosis of VUR than other invasive methods.


Image reconstruction in optical tomography.
Optical tomography is a new medical imaging modality that is at the threshold of realization. A large amount of clinical work has shown the very real benefits that such a method could provide. At the same time a considerable effort has been put into theoretical studies of its probable success. At present there exist gaps between these two realms. In this paper we review some general approaches to inverse problems to set the context for optical tomography, defining both the terms forward problem and inverse problem. An essential requirement is to treat the problem in a nonlinear fashion, by using an iterative method. This in turn requires a convenient method of evaluating the forward problem, and its derivatives and variance. Photon transport models are described for obtaining analytical and numerical solutions for the most commonly used ones are reviewed. The inverse problem is approached by classical gradient-based solution methods. In order to develop practical implementations of these methods, we discuss the important topic of photon measurement density functions, which represent the derivative of the forward problem. We show some results that represent the most complex and realistic simulations of optical tomography yet developed. We suggest, in particular, that both time-resolved, and intensity-modulated systems can reconstruct variations in both optical absorption and scattering, but that unmodulated, non-time-resolved systems are prone to severe artefact. We believe that optical tomography reconstruction methods can now be reliably applied to a wide variety of real clinical data. The expected resolution of the method is poor, meaning that it is unlikely that the type of high-resolution images seen in computed tomography or medical resonance imaging can ever be obtained. Nevertheless we strongly expect the functional nature of these images to have a high degree of clinical significance.


A human D1 dopamine receptor gene is located on chromosome 5 at q35.1 and identifies an EcoRI RFLP.
Dopaminergic neurons have been shown to affect voluntary movement, hormone secretion, and emotional tone. Mediating these activities are two receptor subtypes, D1 and D2, which are biochemically and pharmacologically distinct. The D1 subtype, the most abundant form of dopamine receptor in the central nervous system, stimulates adenylate cyclase, modulates D2 receptor activity, regulates neuron growth and differentiation, and mediates several behavioral responses. Recently we reported the cloning of a human D1 dopamine receptor gene (DRD1). High-stringency hybridization of the DRD1 clone to human genomic blots suggests that DRD1 is single copy. When used to probe a Southern blot made with DNAs from a rodent-human somatic cell hybrid panel, DRD1 hybridized to a 6.5-kb EcoRI restriction fragment which was assigned to chromosome 5. Fluorescent in situ hybridization of this gene to human metaphase chromosomes refined the location of DRD1 to 5q35.1. A search for RFLPs associated with DRD1 identified a two-allele EcoRI RFLP, allowing confirmation of DRD1's localization by linkage analysis in Centre d'Etude du Polymorphisme Humain families.


Treatment of cervical stenotic myelopathy: a cost and outcome comparison of laminoplasty versus laminectomy and lateral mass fusion.
Cervical stenotic myelopathy due to spondylosis or ossification of the posterior longitudinal ligament is often treated with laminoplasty or cervical laminectomy (with fusion). The goal of this study was to compare outcomes, radiographic results, complications, and implant costs associated with these 2 treatments. The authors analyzed the records of 56 patients (age range 42–81 years) who were surgically treated for cervical stenosis. Of this group, 30 underwent laminoplasty and 26 underwent laminectomy with fusion. Patients who had cervical kyphosis or spondylolisthesis were excluded. An average of 4 levels were instrumented in the laminoplasty group and 5 levels in the fusion group (p < 0.01). Forty-two percent of the fusions crossed the cervicothoracic junction, but no laminoplasty instrumentation crossed the cervicothoracic junction, and it only reached C-7 in one-third of the cases. Preoperative and postoperative Nurick grades and modified Japanese Orthopaedic Association (mJOA) scores were obtained. Outcomes were also assessed with neck pain visual analog scale (VAS) scores and the Odom outcome criteria. Postoperative length of stay, complications, and implant costs were calculated. The mean duration of follow-up, average patient age, and length of hospital stay were similar for both groups. The mean Nurick scores were also similar in the 2 groups and improved an average of 1.4 points in both (p < 0.01 for preoperative-postoperative comparison in each group). The mean mJOA scores improved 2.7 points in laminoplasty patients and 2.8 points in fusion patients (p < 0.01 for each group). The mean VAS scores for neck pain did not change significantly in the laminoplasty cohort (3.2 ± 2.8 [SD] preoperatively vs 3.4 ± 2.6 postoperatively, p = 0.50). In the fusion cohort, the mean VAS scores improved from 5.8 ± 3.2 to 3.0 ± 2.3 (p < 0.01). Excellent or good Odom outcomes were observed in 76.7% of the patients in the laminoplasty cohort and 80.8% of those in the fusion cohort (p = 0.71). In the fusion group, complications were twice as common and implant costs were nearly 3 times as high as in the laminoplasty group. When cases involving fusions crossing the cervicothoracic junction were excluded, analysis showed similar complication rates in the 2 groups. Patients treated with laminoplasty and patients treated with laminectomy and fusion had similar improvements in Nurick scores, mJOA scores, and Odom outcomes. Patients who underwent fusion typically had higher preoperative neck pain scores, but their neck pain improved significantly after surgery. There was no significant change in the neck pain scores of patients treated with laminoplasty. Our series suggests cervical fusion significantly reduces neck pain in patients with stenotic myelopathy, but that the cost of the implant and rate of reoperation are greater than in laminoplasty.


Immunologic changes during unplanned treatment interruptions of highly active antiretroviral therapy in children with human immunodeficiency virus type 1 infection.
There are few data on the decline of CD4 cells or percentage during unplanned treatment interruptions in HIV-infected children. Data were analyzed on children undergoing interruptions of three or four drug antiretroviral therapy (HAART) in the United Kingdom and Irish Collaborative HIV Paediatric Study and from Sophia Children's Hospital, Rotterdam. Children were included if they had taken HAART continuously for 3 months or longer before interruption and stopped for at least 4 weeks. The effects of CD4 at treatment initiation and of age, CD4, HIV RNA and disease stage at interruption on the CD4 decline during interruption were explored with linear mixed models. Seventy-one children (median age, 7.0 years) had experienced 82 unplanned interruptions of median duration 4.1 (interquartile range, 2.3 to 7.6) months. HAART was restarted after 59 (72%) interruptions (17 with the same and 42 with a new regimen). In children restarting HAART, median CD4% increased toward preinterruption levels by 6 months, when 46% had HIV RNA <400 copies/ml (vs. 15% at interruption). The rate of CD4% decline in 51 children with a median of 3 (range, 2 to 10) CD4 measurements was 0.52% per month (6.2% annually); it was independent of age, CD4 at HAART, preinterruption CD4 or HIV RNA or previous AIDS. Although the average CD4 decline after stopping HAART appears similar to that in adults, large variability suggests that when designing pediatric trials of planned treatment interruptions, interruption length could be determined by time taken for CD4 to decline below a threshold, rather than by imposing interruptions of fixed duration.


Identification of a mutation in the ubiquitin-fold modifier 1-specific peptidase 2 gene, UFSP2, in an extended South African family with Beukes hip dysplasia.
Beukes hip dysplasia (BHD) is an autosomal dominant disorder of variable penetrance that was originally identified in a large South African family of European origin. BHD is characterised by bilateral dysmorphism of the proximal femur, which results in severe degenerative osteoarthropathy. Previous studies mapped the disorder to a 3.34 Mb region on chromosome 4q35. To fine-map the BHD locus and identify the disease-causing mutation by direct sequencing. The linked BHD allele was refined to 1.33 Mb, reducing the number of candidate genes from 25 to 16. Analysis of protein coding and invariant splice-site sequences in three distantly related individuals identified a single-candidate disease-causing variant c.868T>C within exon 8 of the ubiquitin-fold modifier 1 (Ufm1)-specific peptidase 2 gene, UFSP2. The presence of this unique mutation was confirmed in all 17 affected members of the BHD family who were genotyped. The mutation segregated with the BHD phenotype in the extended family with a two-point (single marker) LOD score of 10.4 (θ=0.0 and 80% penetrance). The mutation predicts the substitution of a highly conserved amino acid, p.Tyr290His, in the encoded protein. In vitro functional assays performed using purified recombinant wild-type and mutant UFSP2 protein demonstrated that the BHD mutation abolishes UFSP2-mediated C-terminal cleavage of its substrate, Ufm1. We report a unique UFSP2 mutation that segregates with the BHD phenotype. The predicted amino acid substitution inactivates UFSP2 proteolytic function, thus implicating the ubiquitin-fold modifier 1 cascade in this form of severe hip osteoarthropathy. The facile polymerase chain reaction-based assay we describe could be used to confirm the diagnosis of BHD, or for presymptomatic testing of members of the extended BHD family.


The effect of concomitant ankylosing spondylitis on long-term outcome of patients with inflammatory bowel disease.
The aim of the present study was to compare the demographic features and long-term outcomes of patients with inflammatory bowel disease (IBD) with or without ankylosing spondylitis (AS). Among 1640 IBD (Crohn's disease and ulcerative colitis), 76 patients with IBD+AS were identified. The study group consisted of 76 patients with IBD with synchronous AS. The control group consisted of patients with only IBD, and those were selected according to their registry sequence number being the previous and next case to the diseased case with IBD+AS. The primary endpoint was to compare the rate of intestinal resections between both groups (IBD vs. IBD+AS). Among 76 patients with IBD+AS, 52 (68%) first presented with IBD, 11 (15%) with AS, and the remaining 13 (17%) had both diagnoses at the same time. The mean follow-up time was significantly longer in patients with IBD+AS (43.4 vs. 27.8 months; p=0.01). Twenty-two percent of patients with IBD and 14% of those with IBD+AS had an intestinal resection (p=NS). Biologic and systemic corticosteroid treatments were significantly more common among patients with IBD+AS (32% vs. 7% for biologics, p<0.0001 and 44% vs. 28% for corticosteroids, p=0.042). Age-sex-adjusted regression analysis for both groups disclosed IBD duration as the only independent predictor for resection (R2=0.178; p=0.016). The present study shows that up to 5% of patients with IBD may have AS. Patients with IBD+AS do not have a worse disease outcome than solo patients with IBD.


Using citrate-functionalized TiO2 nanoparticles to study the effect of particle size on zebrafish embryo toxicity.
TiO2 nanoparticles (NPs) are photoactive, potentially producing toxicity in vivo in the presence of sunlight. We have previously demonstrated photodependent toxicity in zebrafish embryos exposed to TiO2 NPs. Here we investigate the effect of particle size on developing zebrafish exposed to 6, 12 and 15 nm citrate-functionalized anatase TiO2 NPs under either simulated sunlight illumination or in the dark. All three sizes of TiO2 NPs caused photo-dependent toxicity. Under simulated sunlight illumination, the acute toxicity of the 6 nm citrate-TiO2 NPs (120 h LC50 of 23.4 mg L(-1)) exceeded that of the 12 and 15 nm citrate-TiO2 NPs. Exposure to 6 nm particles under illumination also caused a higher incidence of developmental defects than the larger particles. These abnormalities included pericardial edema, yolk-sac edema, craniofacial malformation, and opaque yolk. To gain insight into the mechanisms of toxicity, we measured hydroxyl radicals (˙OH) generated by NPs in vitro and reactive oxygen species (ROS) produced in vivo. We found that on a mass basis, smaller particles generated higher levels of ROS both in vitro and in vivo, and the 6 nm citrate-TiO2 NPs induced more oxidative stress than larger particles in the zebrafish embryo. We examined oxidative DNA damage by measuring 8-hydroxydeoxyguanosine in zebrafish exposed to different-sized citrate-TiO2 NPs and found that 6 nm particles caused more DNA damage than did larger particles (12 and 15 nm) under illumination. Our results indicate a photo-dependent toxicity of citrate-TiO2 NPs to zebrafish embryos, with an inverse relationship between particle size and toxicity. Production of more ROS, resulting in more oxidative stress and more DNA damage, represents one possible mechanism of the higher toxicity of smaller citrate-TiO2 NPs. These results highlight the relationship between citrate-TiO2 NP size and toxicity/oxidative stress in developing zebrafish embryos.


3-D nasal cultures: Systems toxicological assessment of a candidate modified-risk tobacco product.
In vitro toxicology approaches have evolved from a focus on molecular changes within a cell to understanding of toxicity-related mechanisms in systems that can mimic the in vivo environment. The recent development of three dimensional (3-D) organotypic nasal epithelial culture models offers a physiologically robust system for studying the effects of exposure through inhalation. Exposure to cigarette smoke (CS) is associated with nasal inflammation; thus, the nasal epithelium is relevant for evaluating the pathophysiological impact of CS exposure. The present study investigated further the application of in vitro human 3-D nasal epithelial culture models for toxicological assessment of inhalation exposure. Aligned with 3Rs strategy, this study aimed to explore the relevance of a human 3-D nasal culture model to assess the toxicological impact of aerosols generated from a candidate modified risk tobacco product (cMRTP), the Tobacco Heating System (THS) 2.2, as compared with smoke generated from reference cigarette 3R4F. A series of experimental repetitions, where multiple concentrations of THS2.2 aerosol and 3R4F smoke were applied, were conducted to obtain reproducible measurements to understand the cellular/molecular changes that occur following exposure. In agreement with "Toxicity Testing in the 21st Century - a Vision and a Strategy", this study implemented a systems toxicology approach and found that for all tested concentrations the impact of 3R4F smoke was substantially greater than that of THS2.2 aerosol in terms of cytotoxicity levels, alterations in tissue morphology, secretion of pro-inflammatory mediators, impaired ciliary function, and increased perturbed transcriptomes and miRNA expression profiles.


Disseminated unicellular green algal infection in two sheep in India.
Unicellular green algae infected and disseminated in two adult Nellore brown rams in India. At necropsy multiple green lesions were observed in the lungs, liver and kidneys of both sheep. Histological examination revealed necrotizing granulomas that contained numerous spheroidal to elliptical organisms ranging from 5-20 micrometers in diameter. These organisms occurred in several reproductive stages: small (5-9 micrometers) single, undifferentiated cells; intermediate (9-15 micrometers) cells undergoing nuclear and cytoplasmic cleavage; and large, mature, endosporulating cells containing two to six or more endospores. In addition, occasional large (20-35 micrometers), round thick-walled forms were among the typical cells. The cytoplasm of the algal cells contained numerous, large, irregularly-shaped and densely packed granules of varying sizes, which were strongly stained by the Gridley fungus, periodic acid-Schiff, and Gomori methenamine silver procedures. Electron microscopic studies revealed chloroplasts in the cytoplasm of these organisms. The chloroplasts consisted of smoothly contoured electron-lucent or electron-dense granules, 0.5 to 3.0 micrometers in diameter, that were either surrounded by or contiguous to two or more tightly appressed membranes or multidisc bands. Some of the alternating membranes appeared to be spirally twisted or associated with several cytoplasmic granules in one plane of section. This is the first record of algal infections in India.


Algorithms for modeling global and context-specific functional relationship networks.
Functional genomics has enormous potential to facilitate our understanding of normal and disease-specific physiology. In the past decade, intensive research efforts have been focused on modeling functional relationship networks, which summarize the probability of gene co-functionality relationships. Such modeling can be based on either expression data only or heterogeneous data integration. Numerous methods have been deployed to infer the functional relationship networks, while most of them target the global (non-context-specific) functional relationship networks. However, it is expected that functional relationships consistently reprogram under different tissues or biological processes. Thus, advanced methods have been developed targeting tissue-specific or developmental stage-specific networks. This article brings together the state-of-the-art functional relationship network modeling methods, emphasizes the need for heterogeneous genomic data integration and context-specific network modeling and outlines future directions for functional relationship networks.


Activation of rat thymocytes selectively upregulates the expression of somatostatin receptor subtype-1.
Somatostatin and other neuropeptides are known to modulate the proliferative capacity of immune cells. In the present study, we investigated the expression of Somatostatin receptor (SSTR) subtypes on rat thymocytes. RT-PCR analysis of fresh thymocytes showed significant levels of transcripts for the SSTR2 whereas transcripts for the SSTR1 and SSTR3 were not detectable. Interestingly, when the thymocytes were activated with low concentration of Phytohemagglutinin and interleukin 1, the transcript for SSTR1 was markedly increased. Lymphokine induced activation of thymocytes selectively upregulated the SSTR1 since, transcripts for SSTR2 remained the same after activation and SSTR3 was not detectable. PCR amplified fragment of SSTR1 from the activated thymocytes showed identical sequence to the rat brain receptor. The physiological significance of the increase of SSTR1 mRNA in thymocytes after activation remains to be elucidated but it may be possible that these two different subsets of receptors (SSTR1 and SSTR2) are involved in the modulation of thymocyte proliferation and differentiation.


Oxalate nephropathy complicating Roux-en-Y Gastric Bypass: an underrecognized cause of irreversible renal failure.
The most common bariatric surgery is Roux-en-Y gastric bypass (RYGB), which has been associated with hyperoxaluria and nephrolithiasis. We report a novel association of RYGB with renal insufficiency as a result of oxalate nephropathy. Eleven cases of oxalate nephropathy after RYGB were identified from the Renal Pathology Laboratory of Columbia University. The clinical features, pathologic findings, and outcomes are described. Patients were predominantly white (72.7%) with a mean age of 61.3 yr. Indications for RYGB included morbid obesity (eight patients) and reconstruction after total gastrectomy for gastric cancer (three patients). All 11 patients had a history of hypertension, and 9 were diabetic. Patients presented with acute renal failure, often superimposed on mild chronic renal insufficiency (n = 7), at a median of 12 mo after RYGB. The mean creatinine at baseline, at discovery of acute renal failure, and at biopsy was 1.5, 5.0, and 6.5 mg/dl, respectively. Renal biopsies revealed diffuse tubular degenerative changes, abundant tubular calcium oxalate deposits, and varying degrees of tubulointerstitial scarring. In addition, seven biopsies had underlying diabetic glomerulosclerosis and two had glomerulosclerosis attributable to obesity and hypertension. Eight of 11 patients rapidly progressed to ESRD and required hemodialysis at a mean of 3.2 wk after renal biopsy. The remaining three patients were left with significant chronic kidney disease. Oxalate nephropathy is an underrecognized complication of RYGB and typically results in rapid progression to ESRD. Patients with pre-existing renal disease may be at higher risk for this complication.


Menstrual Patterns in the First Gynecological Year: A Systematic Review.
Understanding what constitutes a normal menstrual cycle during the first gynecological year (GY1) is a common concern of adolescents and clinicians. However, limited high-quality evidence exists. We aimed to summarize published literature regarding menstrual and ovulatory patterns in GY1. DESIGN, SETTING, PARTICIPANTS, INTERVENTIONS, AND MAIN OUTCOME MEASURES: Electronic databases (MEDLINE, Pre-MEDLINE, Embase, Web of Science, CINAHL, Cochrane Library) were systematically searched from database inception to 2018. Eligible studies described menstrual cycles, symptoms, or validated ovulatory data in healthy adolescents in GY1. Two authors independently screened studies, extracted data, and assessed methodological quality. Twenty-two studies involving more than 2000 adolescents were included. Thirteen recorded menstrual cycle and/or symptom data and 14 measured ovulation. Mean cycle length ranged from 32 to 61 days and decreased throughout GY1. Mean menses length was 4.9 to 5.4 days. Frequent menstrual bleeding was reported in up to 23% of participants, infrequent menstrual bleeding in up to one-third, and "irregular menstrual bleeding" in up to 43%. Dysmenorrhea was reported by 30%-89% of participants. Prevalence of ovulatory cycles identified using luteal phase serum or salivary progesterone or urinary pregnanediol was 0 to 45% and increased throughout GY1. However, all used definitions that would be considered subovulatory in clinical practice. Menstrual and ovulatory patterns in GY1 are diverse and differ from those of adults. A transitional phase of menstrual and ovulatory immaturity is common. However, ovulation, irregular cycles, and dysmenorrhea are not uncommon. As such, safe sexual practice should be advocated and prompt medical management should be accessible.


An outbreak of Shigella dysenteriae in Sweden, May-June 2009, with sugar snaps as the suspected source.
We report an outbreak of Shigella dysenteriae type 2 infections during May-June 2009 in Sweden, involving 47 suspected cases of whom 35 were laboratory-confirmed. The epidemiological investigation based on interviews with the patients pointed at sugar snaps from Kenya as the source. Shigella was not detected in samples of sugar snaps. However, Escherichia coli was confirmed in three of four samples indicating contamination by faecal material. During April to May 2009 outbreaks with Shigella connected to sugar snaps from Kenya were reported from Norway and Denmark. In the three countries trace back of the indicated sugar snaps revealed a complex system with several involved import companies and distributers. In Sweden one wholesale company was identified and connections were seen to the Danish trace back. These three outbreaks question whether the existing international certification and quality standards that are in place to prevent products from contamination by faecal pathogens are strict enough.


Structure of the major antigenic loop of foot-and-mouth disease virus complexed with a neutralizing antibody: direct involvement of the Arg-Gly-Asp motif in the interaction.
The crystal structure of a synthetic peptide representing the major antigenic loop of foot-and-mouth disease virus (FMDV), complexed with the Fab fragment of a neutralizing monoclonal antibody raised against the virus, has been determined at 2.8 A resolution. The peptide shows a high degree of internal structure with a nearly cyclic conformation. The conserved Arg-Gly-Asp motif, involved in the viral attachment of aphtoviruses to cells, participates directly in the interaction with several complementarity determining regions of the antibody molecule. The Arg-Gly-Asp triplet shows the same open turn conformation found in the reduced form of FMDV of another serotype and also in integrin binding proteins. The observed interactions provide a molecular interpretation of the amino acid replacements observed to occur in mutants resistant to neutralization by this antibody. The structure also suggests a number of restrictions to variation within the epitope which are imposed to keep the Arg-Gly-Asp motif in its functional conformation.


Association of Eotaxin gene family with asthma and serum total IgE.
The Eotaxin gene family (Eotaxin1, Eotaxin2 and Eotaxin3) recruits and activates CCR3-bearing cells such as eosinophils, mast cells and Th2 lymphocytes that play a major role in allergic disorders. To date, the effect of polymorphisms of Eotaxin genes on asthma phenotypes has not been thoroughly examined. In our research, we sequenced whole regions of the Eotaxin gene family to identify polymorphisms, which may be involved in the development of asthma and total serum IgE. We have identified 37 SNPs in the Exotaxin gene family (Exotaxin1, 2 and 3), and 17 common polymorphic sites were selected for genotyping in our asthma cohort (n=721). Statistical analysis revealed that the EOT2+1265A>G G* allele showed significantly lower frequency in asthmatics than in normal healthy controls (0.14 versus 0.23, P=0.002), and that distribution of the EOT2+1265A>G G* allele-containing genotypes was also much lower in asthmatics (26.3 versus 40.8%, P=0.003). In addition, a non-synonymous SNP in Eotaxin1, EOT1+123Ala>Thr showed significant association with total serum IgE levels (P=0.002-0.02). The effect of EOT1+123Ala>Thr on total serum IgE appeared in a gene-dose-dependent manner. Our findings suggest that the development of asthma may be associated with EOT2+1265A>G polymorphisms, and the susceptibility to high IgE production may be attributed to the EOT1+123Ala>Thr polymorphism. Eotaxin variation/haplotype information identified in this study might provide valuable insights into strategies for the control of asthma.


Cloning and expression of Aplysia carboxypeptidase D, a candidate prohormone-processing enzyme.
Many peptide hormones in a variety of species are produced from larger precursors by limited proteolysis at basic amino acid-containing sites. The marine mollusc Aplysia has homologs of mammalian peptide-processing enzymes, including furin, prohormone convertase 1 (PC1), PC2, and carboxypeptidase E (CPE). A novel neuronal Aplysia enzyme was recently identified that was most closely related to carboxypeptidase D (CPD; Fan and Nagle, DNA Cell Biol. 15, 937-945, 1996), a second carboxypeptidase thought to be present in the secretory pathway and to contribute to peptide hormone processing. We have identified and cloned multiple overlapping bag-cell neuron cDNAs that encode two proteins that are members of the CPD family. Sequence analyses demonstrate that the longer CPD protein (1446 residues) contains an N-terminal signal peptide and four carboxypeptidase-like domains; the third and fourth domains are not predicted to form active enzymes, as several critical residues are absent. The shorter CPD protein is predicted to contain two active carboxypeptidase-like domains. Northern blot analysis identified a major Aplysia CPD mRNA (5.3 kb) and several smaller minor transcripts in central nervous system tissue. The CPD was purified from Aplysia ovotestis using a method previously developed for mammalian CPD. The purified Aplysia CPD binds antisera raised against regions of the protein encoded by the Aplysia cDNA clone, as well as an antiserum raised against duck CPD. The enzymatic properties of purified Aplysia CPD are generally similar to those of mammalian CPD. Aplysia CPD is a candidate prohormone-processing enzyme that may play a role in the processing of Aplysia prohormones in the secretory pathway.


In vitro and in vivo anti-leukemic activity of the peptidase-potentiated alkylator melflufen in acute myeloid leukemia.
The novel aminopeptidase potentiated alkylating agent melflufen, was evaluated for activity in acute myeloid leukemia in a range of in vitro models, as well as in a patient derived xenograft study. All tested AML cell lines were highly sensitive to melflufen while melphalan was considerably less potent. In the HL-60 cell line model, synergy was observed for the combination of melflufen and cytarabine, an interaction that appeared sequence dependent with increased synergy when melflufen was added before cytarabine. Also, in primary cultures of AML cells from patients melflufen was highly active, while normal PBMC cultures appeared less sensitive, indicating a 7-fold in vitro therapeutic index. Melphalan, on the other hand, was only 2-fold more potent in the AML patient samples compared with PBMCs. Melflufen was equally active against non-malignant, immature CD34+ progenitor cells and a more differentiated CD34+ derived cell population (GM14), whereas the stem cell like cells were less sensitive to melphalan. Finally, melflufen treatment showed significant anti-leukemia activity and increased survival in a patient derived xenograft of AML in mice. In conclusion, melflufen demonstrates high and significant preclinical activity in AML and further clinical evaluation seem warranted in this disease.


Profiling Interfacility Transfers for Hospitalized Pediatric Patients.
The hospital-to-hospital transfer of pediatric patients is a common practice that is poorly understood. To better understand this practice, we examined a national database to profile pediatric interfacility transfers. We used the 2012 Kids' Inpatient Database to examine characteristics of hospitalized pediatric patients (<21 years; excluding pregnancy diagnoses) with a transfer admission source. We performed descriptive statistics to compare patient characteristics, utilization, and hospital characteristics between those admitted by transfer versus routine admission. We constructed a multivariable logistic regression model to identify patient characteristics associated with being admitted by transfer versus routine admission. Of the 5.95 million nonpregnancy hospitalizations in the United States in 2012, 4.4% were admitted by transfer from another hospital. Excluding neonatal hospitalizations, this rate increased to 9.4% of the 2.10 million nonneonatal, nonpregnancy hospitalizations. Eighty-six percent of transfers were to urban teaching hospitals. The most common transfer diagnoses to all hospitals nationally were mood disorder (8.9%), other perinatal conditions (8.7%), prematurity (4.8%), asthma (4.2%), and bronchiolitis (3.8%). In adjusted analysis, factors associated with higher odds of being admitted by transfer included having a neonatal principal diagnosis, male gender, white race, nonprivate insurance, rural residence, higher illness severity, and weekend admission. Interfacility transfers are relatively common among hospitalized pediatric patients. Higher odds of admission by transfer are associated not only with higher illness severity but also with principal diagnosis, insurance status, and race. Further studies are needed to identify the etiologies and clinical impacts of identified transfer differences.


Comparison of laser mode calculations.
In this paper we show that numerical and kernel expansion procedures for solving the laser mode problem do not differ in essence; both convert the integral equation into a matrix equation. Furthermore, the Fox and Li iterative method is shown to be a matrix diagonalization technique. A particular kernel expansion using Gaussian-Hermite functions is discussed, as are matrix diagonalization techniques. Numerical results are compared with other published values. We conclude that the optimum procedure is to use gaussian quadrature numerical integration to convert to a matrix equation and diagonalize the matrix with the computer program ALLMAT. This method is computationally simple and simultaneously determines many modes. Also, it is applicable to unstable and/or tilted mirror resonators with selectively coated reflectors.


The challenge of abstract concepts.
concepts ("freedom") differ from concrete ones ("cat"), as they do not have a bounded, identifiable, and clearly perceivable referent. The way in which abstract concepts are represented has recently become a topic of intense debate, especially because of the spread of the embodied approach to cognition. Within this framework concepts derive their meaning from the same perception, motor, and emotional systems that are involved in online interaction with the world. Most of the evidence in favor of this view, however, has been gathered with regard to concrete concepts. Given the relevance of abstract concepts for higher-order cognition, we argue that being able to explain how they are represented is a crucial challenge that any theory of cognition needs to address. The aim of this article is to offer a critical review of the latest theories on abstract concepts, focusing on embodied ones. Starting with theories that question the distinction between abstract and concrete concepts, we review theories claiming that abstract concepts are grounded in metaphors, in situations and introspection, and in emotion. We then introduce multiple representation theories, according to which abstract concepts evoke both sensorimotor and linguistic information. We argue that the most promising approach is given by multiple representation views that combine an embodied perspective with the recognition of the importance of linguistic and social experience. We conclude by discussing whether or not a single theoretical framework might be able to explain all different varieties of abstract concepts. (PsycINFO Database Record


Action and emotion recognition from point light displays: an investigation of gender differences.
Folk psychology advocates the existence of gender differences in socio-cognitive functions such as 'reading' the mental states of others or discerning subtle differences in body-language. A female advantage has been demonstrated for emotion recognition from facial expressions, but virtually nothing is known about gender differences in recognizing bodily stimuli or body language. The aim of the present study was to investigate potential gender differences in a series of tasks, involving the recognition of distinct features from point light displays (PLDs) depicting bodily movements of a male and female actor. Although recognition scores were considerably high at the overall group level, female participants were more accurate than males in recognizing the depicted actions from PLDs. Response times were significantly higher for males compared to females on PLD recognition tasks involving (i) the general recognition of 'biological motion' versus 'non-biological' (or 'scrambled' motion); or (ii) the recognition of the 'emotional state' of the PLD-figures. No gender differences were revealed for a control test (involving the identification of a color change in one of the dots) and for recognizing the gender of the PLD-figure. In addition, previous findings of a female advantage on a facial emotion recognition test (the 'Reading the Mind in the Eyes Test' (Baron-Cohen, 2001)) were replicated in this study. Interestingly, a strong correlation was revealed between emotion recognition from bodily PLDs versus facial cues. This relationship indicates that inter-individual or gender-dependent differences in recognizing emotions are relatively generalized across facial and bodily emotion perception. Moreover, the tight correlation between a subject's ability to discern subtle emotional cues from PLDs and the subject's ability to basically discriminate biological from non-biological motion provides indications that differences in emotion recognition may - at least to some degree - be related to more basic differences in processing biological motion per se.


Targeted radionuclide therapy.
Targeted radionuclide therapy (TRT) seeks molecular and functional targets within patient tumor sites. A number of agents have been constructed and labeled with beta, alpha, and Auger emitters. Radionuclide carriers spanning a broad range of sizes; e.g., antibodies, liposomes, and constructs such as nanoparticles have been used in these studies. Uptake, in percent-injected dose per gram of malignant tissue, is used to evaluate the specificity of the targeting vehicle. Lymphoma (B-cell) has been the primary clinical application. Extension to solid tumors will require raising the macroscopic absorbed dose by several-fold over values found in present technology. Methods that may effect such changes include multistep targeting, simultaneous chemotherapy, and external sequestration of the agent. Toxicity has primarily involved red marrow so that marrow replacement can also be used to enhance future TRT treatments. Correlation of toxicities and treatment efficiency has been limited by relatively poor absorbed dose estimates partly because of using standard (phantom) organ sizes. These associations will be improved in the future by obtaining patient-specific organ size and activity data with hybrid SPECT/CT and PET/CT scanners.


Development of a new mouse model of acute pancreatitis induced by administration of L-arginine.
The pathogenesis of acute pancreatitis is not fully understood. Experimental animal models that mimic human disease are essential to better understand the pathophysiology of the disease and to evaluate potential therapeutic agents. Given that the mouse genome is known completely and that a large number of strains with various genetic deletions are available, it is advantageous to have multiple reliable mouse models of acute pancreatitis. Presently, there is only one predominant model of acute pancreatitis in mice, in which hyperstimulatory doses of cholecystokinin or its analog caerulein are administered. Therefore, the aim of this study was to develop another mouse model of acute pancreatitis. In this study, C57BL/6 mice were injected intraperitoneally with L-arginine in two doses of 4 g/kg each, 1 h apart. Serum amylase, myeloperoxidase, and histopathology were examined at varying time points after injection to assess injury to the pancreas and lung. We found that injection of L-arginine was followed by significant increases in plasma amylase and pancreatic myeloperoxidase accompanied by marked histopathological changes. The injury to the pancreas was slow to develop and peaked at 72 h. Subsequent to peak injury, the damaged areas contained collagen fibers as assessed by increased Sirius red staining. In contrast, D-arginine or other amino acids did not cause injury to the pancreas. In addition, acute inflammation in the pancreas was associated with lung injury. Our results indicate that administration of L-arginine to mice results in severe acute pancreatitis. This model should help in elucidating the pathophysiology of pancreatitis.