Angiographic Morphometry of Internal Carotid Artery Circulation in Turkish Children.
Knowledge of cerebrovascular morphology is integral in planning neuroendovascular interventions, especially for procedures involving placement of stents, flow diverters or stentrievers. There is insufficient data on angiographic normative values of cerebral circulation in the pediatric age group since angiograms are uncommonly performed in children except for arteriovenous malformations in which arterial dimensions are larger than normal. We aimed to measure the diameters of internal carotid circulation (ICC) arteries on digital subtraction angiograms of pediatric patients and determine the growth trends. This is a retrospective cross-sectional study measurements of ICC arteries of 64 pediatric patients (4-122 months) with retinoblastoma undergoing intra-arterial chemotherapy. Petrous, cavernous, supraclinoid and choroidal segments of internal carotid artery (ICA) and anterior cerebral artery (ACA) diameters had significant correlation with age. Most of the growth was noted in the first 36-48 months of life. Middle cerebral artery (MCA) diameter did not show significant correlation with age. 87% of the adult diameter of the MCA was attained in the first 6 months of life. ICC arteries reached 81% to 99% of adult sizes in the first 48 months of life. On the contrary, the main iliac artery was only 59% of the adult diameter at this age group. Use of current intracranial stents in children appears tolerable due to the growth pattern of ICC arteries. Based on this data, the current armamentarium of intracranial stents or stent-like devices is sufficient to cover the need in the pediatric population.


Ia afferent excitation of motoneurones in the in vitro new-born rat spinal cord is selectively antagonized by kynurenate.
Intracellular recordings from motoneurones in in vitro preparations of new-born rat spinal cord were used to study the sensitivity of the Ia excitatory post-synaptic potential (e.p.s.p.) to antagonists of excitatory amino acids, in order to test whether group Ia primary afferents release L-glutamate, or a similar compound, as a neurotransmitter. The Ia e.p.s.p. was isolated for study by using low intensity stimulation of individual muscle nerves and by the addition to the superfusate of high concentrations of divalent cations which suppressed polysynaptic inputs to the motoneurones. The pattern of convergence of group Ia afferents from homonymous, heteronymous and antagonist muscle nerves onto motoneurones in the new-born rat was similar to that reported in the adult cat spinal cord. Homonymous muscle nerve stimulation evoked the largest amplitude Ia e.p.s.p.s while heteronymous muscle nerve stimulation elicited smaller e.p.s.p.s or had no effect. Stimulation of antagonist muscle nerves resulted in inhibitory post-synaptic potentials (i.p.s.p.). Superfusion of the specific N-methyl-D-aspartate (NMDA) receptor antagonist, 2-amino-5-phosphonovalerate, did not inhibit the Ia e.p.s.p. but did suppress later, polysynaptic components of the response evoked from dorsal roots. Kynurenate was a potent inhibitor of the Ia e.p.s.p. The site of action of kynurenate was examined by observing its effect on synaptic depression and was found to be consistent with a post-synaptic mechanism. Kynurenate selectively blocked the depolarization of motoneurones elicited by L-glutamate and had no effect on the depolarization evoked by carbachol. The selectivity of action of kynurenate was further examined by comparing its effect on the recurrent i.p.s.p. evoked by ventral root stimulation with its effect on the Ia e.p.s.p. The recurrent i.p.s.p. was antagonized by strychnine and dihydro-beta-erythroidine while kynurenate, at a concentration which greatly reduced the Ia e.p.s.p., had no effect. These results suggest that stimulation of group Ia primary afferents evokes the release of L-glutamate, or a similar compound, which activates non-NMDA excitatory amino acid receptors on motoneurones which, in turn, mediate the Ia e.p.s.p.


Efficacy of Burosumab in Adults with X-linked Hypophosphatemia (XLH): A Post Hoc Subgroup Analysis of a Randomized Double-Blind Placebo-Controlled Phase 3 Study.
The anti-fibroblast growth factor 23 monoclonal antibody burosumab corrects hypophosphatemia in adults with X-linked hypophosphatemia (XLH) and improves pain, stiffness, physical function, and fatigue. This post hoc subgroup analysis used data from the 24-week placebo-controlled period of a phase 3 study in 134 adults with XLH (ClinicalTrials.gov NCT02526160), to assess whether the benefits of burosumab are evident in 14 clinically relevant subgroups defined by baseline demographic and functional criteria, including sex, Brief Pain Inventory-short form (BPI-SF) Average And Worst Pain, region, race, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC®) Stiffness, Physical Function and Pain domains and total score, use of opioid/other pain medication, active fractures/pseudo-fractures, and 6-min walk test distance. There were no statistically significant interactions between any of the subgroups and treatment arm for any endpoint. Higher proportions of subjects achieved mean serum phosphate concentration above the lower limit of normal (the primary endpoint) with burosumab than with placebo in all subgroups. For the key secondary endpoints (WOMAC Stiffness and Physical Function; BPI-SF Worst Pain) individual subgroup categories showed improvements with burosumab relative to placebo. For additional efficacy endpoints, burosumab was favored in some subgroups but differences were not significant and confidence intervals were wide. For some endpoints the treatment effect is small at 24 weeks in all subjects. This subgroup analysis shows that burosumab was largely superior to placebo across endpoints in the 14 clinically relevant subgroup variables at 24 weeks and is likely to benefit all symptomatic adults with active XLH.


Use of acoustics to deter bark beetles from entering tree material.
Acoustic technology is a potential tool to protect wood materials and eventually live trees from colonization by bark beetles. Bark beetles such as the southern pine beetle Dendroctonus frontalis, western pine beetle D. brevicomis and pine engraver Ips pini (Coleoptera: Curculionidae) use chemical and acoustic cues to communicate and to locate potential mates and host trees. In this study, the efficacy of sound treatments on D. frontalis, D. brevicomis and I. pini entry into tree materials was tested. Acoustic treatments significantly influenced whether beetles entered pine logs in the laboratory. Playback of artificial sounds reduced D. brevicomis entry into logs, and playback of stress call sounds reduced D. frontalis entry into logs. Sound treatments had no effect on I. pini entry into logs. The reduction in bark beetle entry into logs using particular acoustic treatments indicates that sound could be used as a viable management tool.


Metal-free sequential [3 + 2]-dipolar cycloadditions using cyclooctynes and 1,3-dipoles of different reactivity.
Although metal-free cycloadditions of cyclooctynes and azides to give stable 1,2,3-triazoles have found wide utility in chemical biology and material sciences, there is an urgent need for faster and more versatile bioorthogonal reactions. We have found that nitrile oxides and diazocarbonyl derivatives undergo facile 1,3-dipolar cycloadditions with cyclooctynes. Cycloadditions with diazocarbonyl derivatives exhibited similar kinetics as compared to azides, whereas the reaction rates of cycloadditions with nitrile oxides were much faster. Nitrile oxides could conveniently be prepared by direct oxidation of the corresponding oximes with BAIB, and these conditions made it possible to perform oxime formation, oxidation, and cycloaddition as a one-pot procedure. The methodology was employed to functionalize the anomeric center of carbohydrates with various tags. Furthermore, oximes and azides provide an orthogonal pair of functional groups for sequential metal-free click reactions, and this feature makes it possible to multifunctionalize biomolecules and materials by a simple synthetic procedure that does not require toxic metal catalysts.


Henle fiber layer phase retardation measured with polarization-sensitive optical coherence tomography.
We developed a method based on polarization-sensitive optical coherence tomography (PS-OCT) to quantify the double pass phase retardation (DPPR) induced by Henle fiber layer in three subjects. Measurements of the retina were performed at a mean wavelength of 840 nm using two polarization states that were perpendicular in a Poincaré sphere representation and phase retardation contributions from tissue layers above and below the Henle fiber layer were excluded using appropriately placed reference and measurement points. These points were semi-automatically segmented from intensity data. Using a new algorithm to determine DPPR, the Henle fiber layer in three healthy subjects aged 50-60 years showed elevated DPPR in a concentric ring about the fovea, with an average maximum DPPR for the three subjects of 22.0° (range: 20.4° to 23.0°) occurring at an average retinal eccentricity of 1.8° (range: 1.5° to 2.25°). Outside the ring, a floor of approximately 6.8° was measured, which we show can mainly be attributed to phase noise that is induced in the polarization states. We also demonstrate the method can determine fast axis orientation of the retardation, which is found consistent with the known radial pattern of Henle fibers.


Deep sequencing of large library selections allows computational discovery of diverse sets of zinc fingers that bind common targets.
The Cys2His2 zinc finger (ZF) is the most frequently found sequence-specific DNA-binding domain in eukaryotic proteins. The ZF's modular protein-DNA interface has also served as a platform for genome engineering applications. Despite decades of intense study, a predictive understanding of the DNA-binding specificities of either natural or engineered ZF domains remains elusive. To help fill this gap, we developed an integrated experimental-computational approach to enrich and recover distinct groups of ZFs that bind common targets. To showcase the power of our approach, we built several large ZF libraries and demonstrated their excellent diversity. As proof of principle, we used one of these ZF libraries to select and recover thousands of ZFs that bind several 3-nt targets of interest. We were then able to computationally cluster these recovered ZFs to reveal several distinct classes of proteins, all recovered from a single selection, to bind the same target. Finally, for each target studied, we confirmed that one or more representative ZFs yield the desired specificity. In sum, the described approach enables comprehensive large-scale selection and characterization of ZF specificities and should be a great aid in furthering our understanding of the ZF domain.


Imaging, biomarker and invasive assessment of diffuse left ventricular myocardial fibrosis in atrial fibrillation.
Using cardiovascular magnetic resonance imaging (CMR), it is possible to detect diffuse fibrosis of the left ventricle (LV) in patients with atrial fibrillation (AF), which may be independently associated with recurrence of AF after ablation. By conducting CMR, clinical, electrophysiology and biomarker assessment we planned to investigate LV myocardial fibrosis in patients undergoing AF ablation. LV fibrosis was assessed by T1 mapping in 31 patients undergoing percutaneous ablation for AF. Galectin-3, coronary sinus type I collagen C terminal telopeptide (ICTP), and type III procollagen N terminal peptide were measured with ELISA. Comparison was made between groups above and below the median for LV extracellular volume fraction (ECV), followed by regression analysis. On linear regression analysis LV ECV had significant associations with invasive left atrial pressure (Beta 0.49, P = 0.008) and coronary sinus ICTP (Beta 0.75, P < 0.001), which remained significant on multivariable regression. LV fibrosis in patients with AF is associated with left atrial pressure and invasively measured levels of ICTP turnover biomarker.


Immunofluorescent triple-staining technique to identify sensory nerve endings in human thumb ligaments.
Ligament innervation purportedly plays a critical role in stability, proprioception and pathology of joints with minimal bony constraints. The human thumb carpometacarpal (CMC) joint is such a joint: with a complex saddle configuration and wide circumduction, its constraint is primarily ligamentous and it is prone to osteoarthritis. CMC reconstruction is the most commonly performed arthritis surgery in the upper extremity. Little, however, is known about CMC ligament innervation. We describe a novel triple-staining immunofluorescence technique using the markers for low-affinity neurotrophin receptor p75, the pan-neuronal marker protein gene product (PGP) 9.5 and 4',6'-diamidino-2-phenylindole (DAPI) to simultaneously detect and differentiate between specific sensory nerve endings: the Pacini corpuscles, the Ruffini endings and nerve fascicles. Five primary CMC ligaments (dorsal radial, dorsal central, posterior oblique, anterior oblique and ulnar collateral ligaments) were harvested from 10 fresh-frozen human cadaver hands. Following paraffin sectioning, each ligament was stained using a triple-stain technique and imaged with fluorescence microscopy. Multidimensional acquisition permitted simultaneous capture of images at different wavelengths. Pacini corpuscles were distinguished by their distinct p75 immunoreactive capsules, and Ruffini endings by their overlapping p75 and PGP9.5 immunoreactive dendritic nerve endings. Simultaneous use of PGP9.5, p75 and DAPI immunofluorescence to analyze innervation patterns in human ligaments provides descriptive analysis of staining patterns and receptor structure as well as clues as to the proprioceptive function of CMC ligaments and the joint as a whole. Our novel findings of CMC ligament innervation augment the study of normal and pathological joint mechanics in this joint so prone to osteoarthritis.


Antibody response profiles induced by Plasmodium falciparum glutamate-rich protein in naturally exposed individuals from a Brazilian area endemic for malaria.
The goal of this study was to evaluate the antibody response induced by Plasmodium falciparum glutamate-rich protein (GLURP) in naturally exposed individuals from the Brazilian Amazon region (Rondonia State). The results showed that most individuals had IgG against two well-defined regions within P. falciparum GLURP, the relatively conserved N-terminal nonrepeat region (R0) and the immunodominant repeat region (R2), 67% and 79%, respectively. The peptides S4 from R2 (53%) and P11 from R0 (49%) were identified as immunodominant B cell epitopes and induced higher levels of antibodies. The number of GLURP peptides recognized and the levels of IgG against S4 and P11 peptides showed a positive correlation with age and time of exposure in the malaria-endemic area studied. The antibody responses against GLURP epitopes appear to be modulated by HLA class II antigens. Interestingly, the GLURP immunodominant B cell epitopes in individuals from a Brazilian malaria-endemic area are distinguishable from those of the African malaria-endemic area. Considering the importance of GLURP as a malaria vaccine candidate and the increasing focus on the use of subunit vaccines in the control of infectious diseases, the concern of the influence of class II allele frequencies in ethnically diverse populations may be important before vaccine trials are conducted among people naturally exposed to malaria parasites.


What is the cost of diagnosis and management of drug resistant tuberculosis in South Africa?
Drug-resistant tuberculosis (DR-TB) is undermining TB control in South Africa. However, there are hardly any data about the cost of treating DR-TB in high burden settings despite such information being quintessential for the rational planning and allocation of resources by policy-makers, and to inform future cost-effectiveness analyses. We analysed the comparative 2011 United States dollar ($) cost of diagnosis and treatment of drug sensitive TB (DS-TB), MDR-TB and XDR-TB, based on National South African TB guidelines, from the perspective of the National TB Program using published clinical outcome data. Assuming adherence to national DR-TB management guidelines, the per patient cost of XDR-TB was $26,392, four times greater than MDR-TB ($6772), and 103 times greater than drug-sensitive TB ($257). Despite DR-TB comprising only 2.2% of the case burden, it consumed ~32% of the total estimated 2011 national TB budget of US $218 million. 45% and 25% of the DR-TB costs were attributed to anti-TB drugs and hospitalization, respectively. XDR-TB consumed 28% of the total DR-TB diagnosis and treatment costs. Laboratory testing and anti-TB drugs comprised the majority (71%) of MDR-TB costs while hospitalization and anti-TB drug costs comprised the majority (92%) of XDR-TB costs. A decentralized XDR-TB treatment programme could potentially reduce costs by $6930 (26%) per case and reduce the total amount spent on DR-TB by ~7%. Although DR-TB forms a very small proportion of the total case burden it consumes a disproportionate and substantial amount of South Africa's total annual TB budget. These data inform rational resource allocation and selection of management strategies for DR-TB in high burden settings.


A small molecule TrkB ligand reduces motor impairment and neuropathology in R6/2 and BACHD mouse models of Huntington's disease.
Loss of neurotrophic support in the striatum caused by reduced brain-derived neurotrophic factor (BDNF) levels plays a critical role in Huntington's disease (HD) pathogenesis. BDNF acts via TrkB and p75 neurotrophin receptors (NTR), and restoring its signaling is a prime target for HD therapeutics. Here we sought to determine whether a small molecule ligand, LM22A-4, specific for TrkB and without effects on p75(NTR), could alleviate HD-related pathology in R6/2 and BACHD mouse models of HD. LM22A-4 was administered to R6/2 mice once daily (5-6 d/week) from 4 to 11 weeks of age via intraperitoneal and intranasal routes simultaneously to maximize brain levels. The ligand reached levels in the R6/2 forebrain greater than the maximal neuroprotective dose in vitro and corrected deficits in activation of striatal TrkB and its key signaling intermediates AKT, PLCγ, and CREB. Ligand-induced TrkB activation was associated with a reduction in HD pathologies in the striatum including decreased DARPP-32 levels, neurite degeneration of parvalbumin-containing interneurons, inflammation, and intranuclear huntingtin aggregates. Aggregates were also reduced in the cortex. Notably, LM22A-4 prevented deficits in dendritic spine density of medium spiny neurons. Moreover, R6/2 mice given LM22A-4 demonstrated improved downward climbing and grip strength compared with those given vehicle, though these groups had comparable rotarod performances and survival times. In BACHD mice, long-term LM22A-4 treatment (6 months) produced similar ameliorative effects. These results support the hypothesis that targeted activation of TrkB inhibits HD-related degenerative mechanisms, including spine loss, and may provide a disease mechanism-directed therapy for HD and other neurodegenerative conditions.


Carrying capacity and demographic stochasticity: scaling behavior of the stochastic logistic model.
The stochastic logistic model is the simplest model that combines individual-level demography with density dependence. It explicitly or implicitly underlies many models of biodiversity of competing species, as well as non-spatial or metapopulation models of persistence of individual species. The model has also been used to study persistence in simple disease models. The stochastic logistic model has direct relevance for questions of limiting similarity in ecological systems. This paper uses a biased random walk heuristic to derive a scaling relationship for the persistence of a population under this model, and discusses its implications for models of biodiversity and persistence. Time to extinction of a species under the stochastic logistic model is approximated by the exponential of the scaling quantity U=(R-1)(2) N/R(R+1), where N is the habitat size and R is the basic reproductive number.


Granulocyte and Monocyte Apheresis for Induction of Remission in Children With New-Onset Inflammatory Bowel Colitis.
The aim of the study was to analyze the effect of granulocyte and monocyte apheresis (GMA) with mesalazine for induction of remission in pediatric patients with newly onset chronic inflammatory bowel disease (IBD) colitis. Thirteen pediatric patients with newly onset extensive IBD colitis were investigated per the ECCO/ESPGHAN IBD protocol. Of these 13, 12 received 10 treatments with Adacolumn (ADA) during a median of 6.25 weeks in combination with low-to-moderate doses of mesalazine, which was continued after apheresis. A control colonoscopy was performed 12 to 16 weeks after GMA treatment. Primary outcomes were mucosal healing (Mayo endoscopic score) and histopathologic grading of biopsies. A secondary outcome was disease activity as measured by the Pediatric Ulcerative Colitis Activity Index. Twelve children (6 girls) with a median age of 14.6 years and a median duration of symptoms at diagnosis of 3.2 months received all planned 10 treatment sessions with ADA. Ten of 12 patients had pancolitis and 2 of 12 extensive colitis. A final diagnosis, however, indicated ulcerative colitis in 10 children and Crohn disease in 2 children. At control colonoscopy, 8 of 12 children were in clinical remission and the Mayo endoscopic score showed significant improvement in 9 of 12 patients (P = 0.006). Complete microscopic remission, according to the Geboes score, was observed in 2 patients. In this small study GMA for induction of remission of newly onset pediatric IBD colitis was effective in 8 of 12 patients. Further controlled studies are warranted to confirm the efficacy of this treatment model.


Optimizing hepcidin measurement with a proficiency test framework and standardization improvement.
Hepcidin measurement advances insights in pathophysiology, diagnosis, and treatment of iron disorders, but requires analytically sound and standardized measurement procedures (MPs). Recent development of a two-level secondary reference material (sRM) for hepcidin assays allows worldwide standardization. However, no proficiency testing (PT) schemes to ensure external quality assurance (EQA) exist and the absence of a high calibrator in the sRM set precludes optimal standardization. We developed a pilot PT together with the Dutch EQA organization Stichting Kwaliteitsbewaking Medische Laboratoriumdiagnostiek (SKML) that included 16 international hepcidin MPs. The design included 12 human serum samples that allowed us to evaluate accuracy, linearity, precision and standardization potential. We manufactured, value-assigned, and validated a high-level calibrator in a similar manner to the existing low- and middle-level sRM. The pilot PT confirmed logistical feasibility of an annual scheme. Most MPs demonstrated linearity (R2>0.99) and precision (duplicate CV>12.2%), although the need for EQA was shown by large variability in accuracy. The high-level calibrator proved effective, reducing the inter-assay CV from 42.0% (unstandardized) to 14.0%, compared to 17.6% with the two-leveled set. The calibrator passed international homogeneity criteria and was assigned a value of 9.07±0.24 nmol/L. We established a framework for future PT to enable laboratory accreditation, which is essential to ensure quality of hepcidin measurement and its use in patient care. Additionally, we showed optimized standardization is possible by extending the current sRM with a third high calibrator, although international implementation of the sRM is a prerequisite for its success.


Derivation and Application of Molecular Signatures to Prostate Cancer: Opportunities and Challenges.
Prostate cancer is a high-incidence cancer that requires improved patient stratification to ensure accurate predictions of risk and treatment response. Due to the significant contributions of transcription factors and epigenetic regulators to prostate cancer progression, there has been considerable progress made in developing gene signatures that may achieve this. Some of these are aligned to activities of key drivers such as the androgen receptor, whilst others are more agnostic. In this review, we present an overview of these signatures, the strategies for their derivation, and future perspectives on their continued development and evolution.


MyD88 or TRAM knockdown regulates interleukin (IL)-6, IL-8, and CXCL12 mRNA expression in human gingival and periodontal ligament fibroblasts.
In a previous report, it was shown that Toll-like receptor (TLR) 2 knockdown modulates interleukin (IL)-6 and IL-8 but not the chemokine CXCL12, an important mediator with inflammatory and proangiogenic effects, in human gingival fibroblasts (HGF) and human periodontal ligament fibroblasts (HPDLF). This study investigates whether knocking down two important TLR adaptor molecules, such as myeloid differentiation protein 88 (MyD88) and TRIF-related adaptor molecule (TRAM), could affect mRNA expression of IL-6, IL-8, and CXCL12 in HGF and HPDLF. After small interfering (si) RNA-mediated silencing of MyD88 or TRAM, HGF and HPDLF were stimulated with Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS) or two synthetic ligands of TLR2 (Pam2CSK4 and Pam3CSK4) for 6 hours. IL-6, IL-8, and CXCL12 mRNAs were evaluated by quantitative polymerase chain reaction. Knockdown of MyD88 or TRAM partially impaired the IL-8 mRNA upregulation in both fibroblast subpopulations. Similarly, IL-6 upregulation was partially prevented by siMyD88 or siTRAM in HGF stimulated with Pg LPS, as well as in both fibroblast subtypes challenged with Pam2CSK4. Conversely, constitutive CXCL12 mRNA levels were upregulated by MyD88 or TRAM knockdown in non-stimulated cells. These results suggest that TLR adaptor molecules knockdown, such as MyD88 or TRAM, can decrease IL-6 and IL-8 mRNA and increase CXCL12 mRNA expression in HGF and HPDLF. This can be an important step for better understanding the mechanisms that control the inflammatory cytokine and chemokine expression, which in turn contributes to periodontal pathogenesis.


Mitochondrial and redox dysfunction in post-menopause as risk factor of neurodegenerative disease: a pilot study testing the role of a validated Japanese functional food.
During the menopause women may experience increased oxidative stress and decreased antioxidant capacity and, together with the decline of neurosteroids, this represents a risk factor for Alzheimer's disease. The aim of the present study was to test a functional food (FPP-ORI, Osato Research Institute, Gifu, Japan) on redox and mitochondrial efficiency in post-menopausal women. The study population consisting of 69 untreated post-menopausal women were given supplements as follows: Group A was given a multivitamin (MV) 1c 2 times a day, and group B was given FPP 4.5 g 2 times a day. Group C consisted of 23 fertile premenopausal women as the control group. The tests carried out on entry, and at 3 and 6 months were erythrocyte redox parameters, plasma oxidated proteins, brain-derived neurotrophic factor (BDNF) and peripheral blood mononuclear cell (PBMC) mitochondria cytochrome c oxidase Vmax activity. Menopausal women showed an increased malondialdehyde (MDA) (p<0.05 vs control) which was normalized by both treatments (p<0.05), but MV failed to do so in the BMI ≥26 subgroup (p<0.05). All other redox enzymes and BDNF were significantly lower in menopausal women and they responded only to FPP (p<0.05). Carbonyl protein level was higher in "BMI ≥ 26" subgroup (p<0.05) and reduced only by FPP (p<0.05). The PBMC cyclooxygenase to citrate synthase activity was reduced (<40%) in the menopausal group (p<0.01) and only FPP caused a significant restoration (p<0.05). Although preliminary, these data confirm the redox and mitochondrial dysfunction occurring in post-menopause and responsive to FPP but very poorly to high dosage antioxidants. This may lead to potential preventive opportunities in menopause-associated neurodegenerative disease.


Blocking protein geranylgeranylation is essential for lovastatin-induced apoptosis of human acute myeloid leukemia cells.
Lovastatin is an inhibitor of the enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the major regulatory enzyme of the mevalonate pathway. We have previously reported that lovastatin induces a significant apoptotic response in human acute myeloid leukemia (AML) cells. To identify the critical biochemical mechanism(s) essential for lovastatin-induced apoptosis, add-back experiments were conducted to determine which downstream product(s) of the mevalonate pathway could suppress this apoptotic response. Apoptosis induced by lovastatin was abrogated by mevalonate (MVA) and geranylgeranyl pyrophosphate (GGPP), and was partially inhibited by farnesyl pyrophosphate (FPP). Other products of the mevalonate pathway including cholesterol, squalene, lanosterol, desmosterol, dolichol, dolichol phosphate, ubiquinone, and isopentenyladenine did not affect lovastatin-induced apoptosis in AML cells. Our results suggest that inhibiting geranylgeranylation of target proteins is the predominant mechanism of lovastatin-induced apoptosis in AML cells. In support of this hypothesis, the geranylgeranyl transferase inhibitor (GGTI-298) mimicked the effect of lovastatin, whereas the farnesyl transferase inhibitor (FTI-277) was much less effective at triggering apoptosis in AML cells. Inhibition of geranylgeranylation was monitored and associated with the apoptotic response induced by lovastatin and GGTI-298 in the AML cells. We conclude that blockage of the mevalonate pathway, particularly inhibition of protein geranylgeranylation holds a critical role in the mechanism of lovastatin-induced apoptosis in AML cells.


Neutrophil serine proteases in antibacterial defense.
Neutrophil serine proteases (NSPs) are critical for the effective functioning of neutrophils and greatly contribute to immune protection against bacterial infections. Thanks to their broad substrate specificity, these chymotrypsin-like proteases trigger multiple reactions that are detrimental to bacterial survival such as direct bacterial killing, generation of antimicrobial peptides, inactivation of bacterial virulence factors and formation of neutrophil extracellular traps. Recently, the importance of NSPs in antibacterial defenses has been further underscored by discoveries of unique bacterial evasion strategies to combat these proteases. Bacteria can indirectly disarm NSPs by protecting bacterial substrates against NSP cleavage, but also produce inhibitory molecules that potently block NSPs. Here we review recent insights in the functional contribution of NSPs in host protection against bacterial infections and the elegant strategies that bacteria use to counteract these responses.


The genome sequence of Methanohalophilus mahii SLP(T) reveals differences in the energy metabolism among members of the Methanosarcinaceae inhabiting freshwater and saline environments.
Methanohalophilus mahii is the type species of the genus Methanohalophilus, which currently comprises three distinct species with validly published names. Mhp. mahii represents moderately halophilic methanogenic archaea with a strictly methylotrophic metabolism. The type strain SLP(T) was isolated from hypersaline sediments collected from the southern arm of Great Salt Lake, Utah. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 2,012,424 bp genome is a single replicon with 2032 protein-coding and 63 RNA genes and part of the Genomic Encyclopedia of Bacteria and Archaea project. A comparison of the reconstructed energy metabolism in the halophilic species Mhp. mahii with other representatives of the Methanosarcinaceae reveals some interesting differences to freshwater species.


[Infestation of an urban area by Aedes aegypti and relation with socioeconomic levels].
To assess the association between Aedes aegypti larvae infestation rates and socioeconomic factors. Infestation rates in the urban area of the city of São José do Rio Preto, Southeastern Brazil, were calculated in January of 2005 and subsequently geocoded by address. The urban census tracts were grouped by means of main component analysis, thus producing four socioeconomic clusters (1-4, in a decreasing order of socioeconomic level) and a fifth cluster (5) with a level below the others and not belonging to the urban census tracts (new districts and irregular residential development projects). The Breteau index (BI), the house index (HI), and the container index (CI), as well as the averages of existing containers surveyed per home, were calculated for each cluster. The values of infestation indices did not show significant differences among socioeconomic clusters 1 to 4, even though they were lower than the indices obtained for cluster 5. The averages of existing containers surveyed were higher for cluster 1 in relation to clusters 2 to 4, but did not show significant differences when compared to cluster 5. Larval indices did not show association with the different socioeconomic levels of the cluster that corresponds to the urban census tracts. Nonetheless, new districts, irregular residential development projects and locations adjacent to those with the worst basic sanitary conditions showed the highest values for these indicators.


A critical role of two positively charged amino acids in the Jas motif of Arabidopsis JAZ proteins in mediating coronatine- and jasmonoyl isoleucine-dependent interactions with the COI1 F-box protein.
Coronatine is an important virulence factor produced by several pathovars of the bacterial pathogen Pseudomonas syringae. The structure of coronatine is similar to that of a class of plant hormones called jasmonates (JAs). An important step in JA signaling is the SCF(COI1) E3 ubiquitin ligase-dependent degradation of JAZ repressor proteins. We have recently shown that jasmonoyl isoleucine (JA-Ile) promotes physical interaction between Arabidopsis JAZ1 and COI1 (the F-box component of SCF(COI1)) proteins, and that the JA-Ile-dependent COI1-JAZ1 interaction could be reconstituted in yeast cells (i.e. in the absence of other plant proteins). Here we show that coronatine, but not its two biosynthetic precursors, also promotes interaction between Arabidopsis COI1 and multiple JAZ proteins. The C-terminal Jas motif, but not the N-terminal (NT) domain or central ZIM domain of JAZ proteins, is critical for JA-Ile/coronatine-dependent interaction with COI1. Two positively charged amino acid residues in the Jas domain were identified as essential for coronatine-dependent COI1-JAZ interactions. Mutations of these two residues did not affect the ability of JAZ1 and JAZ9 to interact with the transcription factor AtMYC2. Importantly, transgenic Arabidopsis plants expressing JAZ1 carrying these two mutations exhibited JA-insensitive phenotypes, including male sterility and enhanced resistance to P. syringae infection. These results not only suggest that coronatine and JA-Ile target the physical interaction between COI1 and the Jas domain of JAZ repressors, but also illustrate the critical role of positively charged amino acids in the Jas domain in mediating the JA-Ile/coronatine-dependent JAZ interaction with COI1.


Using Mendelian Randomization to Decipher Mechanisms of Bone Disease.
This review summarizes the basic principles of Mendelian randomization (MR) and provides evidence for the causal effect of multiple modifiable factors on bone outcomes. Several studies using MR approach have provided support for the causal effect of obesity on bone mineral density (BMD). Strikingly, studies have failed to prove a causal association between elevated 25(OH) D concentrations and higher BMD in community-dwelling individuals. The MR approach has been successfully used to evaluate multiple factors related to bone mineral density variation and/or fracture risk. The MR approach avoids some of the classical observational study limitations and provides more robust causal evidence, ensuring bigger success of the clinical trials. The selection of interventions based on genetic evidence could have a substantial impact on clinical practice.


A return to diploma-prepared registered nurses.
A quarter of a century ago, the nursing profession in Canada took a bold step: to establish baccalaureate preparation in nursing as the standard for practice as a registered nurse in Canada. Towards that end, hospital schools of nursing gave way to collaborative programs designed to help hospital and college diploma nursing students make the transition into baccalaureate nursing programs. It is thus paradoxical that, today, the nursing profession in Canada should do anything to jeopardize its goal to have RNs prepared at the baccalaureate level. The purpose of this paper is to focus Canadian nurse leaders' attention on that very possibility, given that some professional nursing organizations in Canada have conceptualized a future RN practice that could result in the emergence of a new kind of healthcare professional and a return to diploma preparation of RNs.


Graphene-extracted membrane lipids facilitate the activation of integrin αvβ8.
Despite the remarkable electrochemical properties of graphene, strong van der Waals attraction between graphene and biomolecules often causes cytotoxicity, which hinders its applications in the biomedical field. Unfortunately, surface passivation of graphene might stimulate undesired immune response as the nanomaterial triggers cytokine production through membrane receptor activation. Herein, we use all-atom Molecular Dynamics (MD) simulations to unravel the underlying mechanism of graphene-induced inside-out activation of integrin αvβ8, a prominent membrane receptor expressed in immune cells. We model the transmembrane (TM) domains of integrin αvβ8 in a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) lipid bilayer and observe the structural changes in the integrin-membrane complex when interacting with a graphene nanosheet across the membrane. We find that the β8 TM domain interacts with the graphene nanosheet directly or indirectly through extracted lipids, facilitating the pulling of a β8 subunit away from an αv subunit and thus leading to the disruption of the TM domain association by breaking the hydrophobic cluster in the cytoplasmic domains of the αv and β8 subunits. Alanine substitution of two conserved phenylalanine residues on the αv subunit at this hydrophobic cluster further reveals the importance of a stable T-shaped structure in retaining integrin in its inactive state. Our results agree with previous studies on the interactions between other integrin subtypes and their endogenous activators, suggesting an intriguing role that the graphene nanosheet may play in the integrin-related signal transduction during its interaction with the membrane.


Clearance of 99mTc DTPA from guinea pig nasal, tracheobronchial, and bronchoalveolar airways.
Technetium-99m labelled diethylenetriamine penta-acetate (99mTc-DTPA) was used to compare small solute absorption (clearance) from nasal, tracheobronchial, and bronchoalveolar airways in anaesthetised guinea pigs. 99mTc DTPA dissolved in saline was superfused through nasal and orolaryngeal catheters on to nasal and tracheobronchial airways; a small particle aerosol of nebulised 99mTc DTPA was delivered to the bronchoalveolar airways through a tracheostomy. Radioactivity over the appropriate region was then determined with a gamma camera. Mucociliary transport of 99mTc DTPA appeared not to contribute to the disappearance of 99mTc DTPA. Time-activity curves were obtained and half life values calculated by fitting a monoexponential equation to the experimental data. A progressive reduction in 99mTc DTPA was recorded from the nasal and tracheobronchial airways and from the bronchoalveolar airway, suggesting that absorption was occurring. The disappearance of 99mTc DTPA was fastest from the bronchoalveolar region, which also had the largest mucosal surface. The similar shape of the retention curves for the nasal and tracheobronchial regions suggests that the characteristics of nasal absorption of 99mTc DTPA could prove applicable to the tracheobronchial region. It is proposed that the present methods are suited for comparing the pharmacology of small solute absorption across nasal, tracheobronchial, and bronchoalveolar airway mucosa.


Proteomics in uveal melanoma research: opportunities and challenges in biomarker discovery.
Uveal melanoma (UM) is the most frequent primary intraocular tumor in adult humans. Despite the significant advances in diagnosis and treatment of UM in the last decades, the prognosis of UM sufferers is still poor. Metastatic liver disease is the leading cause of death in UM and can develop after a long disease-free interval, suggesting the presence of occult micrometastasis. Proteomics technology has opened new opportunities for elucidating the molecular mechanism of complex diseases, such as cancer. This article will review the recent developments in biomarker discovery for UM research by proteomics. In the last few years, the first UM proteomics-based analyses have been launched, yielding promising results. An update on recent developments on this field is presented.


Retinal vessel segmentation: an efficient graph cut approach with retinex and local phase.
Our application concerns the automated detection of vessels in retinal images to improve understanding of the disease mechanism, diagnosis and treatment of retinal and a number of systemic diseases. We propose a new framework for segmenting retinal vasculatures with much improved accuracy and efficiency. The proposed framework consists of three technical components: Retinex-based image inhomogeneity correction, local phase-based vessel enhancement and graph cut-based active contour segmentation. These procedures are applied in the following order. Underpinned by the Retinex theory, the inhomogeneity correction step aims to address challenges presented by the image intensity inhomogeneities, and the relatively low contrast of thin vessels compared to the background. The local phase enhancement technique is employed to enhance vessels for its superiority in preserving the vessel edges. The graph cut-based active contour method is used for its efficiency and effectiveness in segmenting the vessels from the enhanced images using the local phase filter. We have demonstrated its performance by applying it to four public retinal image datasets (3 datasets of color fundus photography and 1 of fluorescein angiography). Statistical analysis demonstrates that each component of the framework can provide the level of performance expected. The proposed framework is compared with widely used unsupervised and supervised methods, showing that the overall framework outperforms its competitors. For example, the achieved sensitivity (0:744), specificity (0:978) and accuracy (0:953) for the DRIVE dataset are very close to those of the manual annotations obtained by the second observer.


Design and analysis of a model predictive controller for active queue management.
Model predictive (MP) control as a novel active queue management (AQM) algorithm in dynamic computer networks is proposed. According to the predicted future queue length in the data buffer, early packets at the router are dropped reasonably by the MPAQM controller so that the queue length reaches the desired value with minimal tracking error. The drop probability is obtained by optimizing the network performance. Further, randomized algorithms are applied to analyze the robustness of MPAQM successfully, and also to provide the stability domain of systems with uncertain network parameters. The performances of MPAQM are evaluated through a series of simulations in NS2. The simulation results show that the MPAQM algorithm outperforms RED, PI, and REM algorithms in terms of stability, disturbance rejection, and robustness.


Composite Bioscaffolds Incorporating Decellularized ECM as a Cell-Instructive Component Within Hydrogels as In Vitro Models and Cell Delivery Systems.
Decellularized tissues represent promising biomaterials, which harness the innate capacity of the tissue-specific extracellular matrix (ECM) to direct cell functions including stem cell proliferation and lineage-specific differentiation. However, bioscaffolds derived exclusively from decellularized ECM offer limited versatility in terms of tuning biomechanical properties, as well as cell-cell and cell-ECM interactions that are important mediators of the cellular response. As an alternative approach, in the current chapter we describe methods for incorporating cryo-milled decellularized tissues as a cell-instructive component within a hydrogel carrier designed to crosslink under mild conditions. This composite strategy can enable in situ cell encapsulation with high cell viability, allowing efficient seeding with a homogeneous distribution of cells and ECM. Detailed protocols are provided for the effective decellularization of human adipose tissue and porcine auricular cartilage, as well as the cryo-milling process used to generate the ECM particles. Further, we describe methods for synthesizing methacrylated chondroitin sulphate (MCS) and for performing UV-initiated and thermally induced crosslinking to form hydrogel carriers for adipose and cartilage regeneration. The hydrogel composites offer great flexibility, and the hydrogel phase, ECM source, particle size, cell type(s) and seeding density can be tuned to promote the desired cellular response. Overall, these systems represent promising platforms for the development of tissue-specific 3-D in vitro cell culture models and in vivo cell delivery systems.


Evidence for involvement of ventral tegmental area cyclic AMP systems in behavioral sensitization to psychostimulants.
The present study investigated the role of ventral tegmental area (VTA) cyclic AMP (cAMP) systems in the behavioral sensitivity to psychostimulants in male Sprague-Dawley rats. Bilateral microinjections of cholera toxin (CTX) into the VTA (50-500 ng/500 nl/side) dose-dependently sensitized animals to the locomotor stimulant effects of systemic d-amphetamine, cocaine and apomorphine, but were without effects on morphine-induced locomotion 24 hr after microinjection. The CTX-induced behavioral sensitization to amphetamine was even greater 72 hr after microinjection, but was no longer present 14 days after intra-VTA CTX pretreatment. Coadministration of the cAMP-dependent protein kinase inhibitor H8 into the VTA blocked CTX-induced sensitization to amphetamine, suggesting that the sensitization is dependent on phosphorylation events in the VTA mediated by cAMP-dependent protein kinase. Pretreatment with CTX did not enhance amphetamine-induced dopamine release in the nucleus accumbens relative to saline controls 24 hr after microinjection. A single bilateral injection of d-amphetamine into the VTA (5 micrograms/side) produced a significant sensitization to systemic amphetamine challenge 72 hr later, and this effect was also blocked by coadministration of H8 into the VTA. These results extend previous studies which have established the importance of the VTA in the development of behavioral sensitization and suggest that cAMP systems may play a crucial role in this neuroadaptive process.


Controversies in the management of alcoholic liver disease.
Alcohol is a risk factor for chronic disease burden in developed countries. Alcoholic liver disease affects 1% of the North American population and is the second most frequent indication for liver transplantation in the United States. It is a spectrum that ranges from simple hepatic steatosis to alcoholic hepatitis to steatohepatitis and eventually cirrhosis. The clinical spectrum of alcoholic hepatitis is wide and ranges from the asymptomatic patient to overt liver failure and death. Liver biopsy as a means of prognostication in alcoholic hepatitis has mostly been replaced with less invasive scoring systems. The management of alcoholic liver disease is challenging. Abstinence is the cornerstone of therapy and should include rehabilitation with a multidisciplinary approach. No specific treatment is required in mild to moderate alcoholic hepatitis. In patients with severe hepatitis, there appears to be a moderate survival benefit from the use of either corticosteroids or pentoxifylline in the absence of contraindications to their use. Nonresponders should have steroid therapy withdrawn by day 7, as persistence with therapy is not beneficial. Orthotopic liver transplantation remains the definitive therapy for decompensated alcoholic cirrhosis despite alcohol abstinence. More studies are needed to define the optimal timing of orthotopic liver transplantation and patients at risk of alcohol relapse post-transplant. Mt Sinai J Med 76:484-498, 2009. (c) 2009 Mount Sinai School of Medicine.


VET: Vasculature Extraction Transformer for Single-Scan Optical Coherence Tomography Angiography.
Optical coherence tomography angiography (OCTA) is a non-invasive imaging modality for analyzing skin microvasculature, enabling non-invasive diagnosis and treatment monitoring. Traditional OCTA algorithms necessitate at least two-repeated scans to generate microvasculature images, while image quality is highly dependent on the repetitions of scans (e.g., 4-8). Nevertheless, a higher repetition count increases data acquisition time, causing patient discomfort and more unpredictable motion artifacts, which can result in potential misdiagnosis. To address these limitations, we proposed a vasculature extraction pipeline based on the novelty vasculature extraction transformer (VET) to generate OCTA images by using a single OCT scan. Distinct from the vision Transformer, VET utilizes convolutional projection to better learn the spatial relationships between image patches. This study recruited 15 healthy participants. The OCT scans were performed in five various skin sites, i.e., palm, arm, face, neck, and lip. Our results show that in comparison to OCTA images obtained by the speckle variance OCTA (peak-signal-to-noise ratio (PSNR): 16.13) and eigen-decomposition OCTA (PSNR: 17.08) using four repeated OCT scans, OCTA images extracted by the proposed pipeline exhibit a better PSNR (18.03) performance while reducing the data acquisition time by 75%. Visual comparisons show that the proposed pipeline outperformed traditional OCTA algorithms, particularly in the imaging of lip and face areas, where artifacts are commonly encountered. This study is the first to demonstrate that the VET can efficiently extract high-quality vasculature images from a single, rapid OCT scan. This capability significantly enhances diagnostic accuracy for patients and streamlines the imaging process.


Electrophysiological correlates of figure-ground segregation directly reflect perceptual saliency.
In a figure identification task, we investigated the influence of different visual cue configurations (spatial frequency, orientation or a combination of both) on the human EEG. Combining psychophysics with ERP and time-frequency analysis, we show that the neural response at about 200ms reflects perceptual saliency rather than physical cue contrast. Increasing saliency caused (i) a negative shift of the posterior P2 coinciding with a power decrease in the posterior theta-band and (ii) an amplitude and latency increase of the posterior P3. We demonstrate that visual cues interact for a percept that is non-linearly related to the physical figure-ground properties.


Distribution and evolution of nitrogen fixation genes in the phylum Bacteroidetes.
Diazotrophs had not previously been identified among bacterial species in the phylum Bacteroidetes until the rapid expansion of bacterial genome sequences, which revealed the presence of nitrogen fixation (nif) genes in this phylum. We herein determined the draft genome sequences of Bacteroides graminisolvens JCM 15093(T) and Geofilum rubicundum JCM 15548(T). In addition to these and previously reported 'Candidatus Azobacteroides pseudotrichonymphae' and Paludibacter propionicigenes, an extensive survey of the genome sequences of diverse Bacteroidetes members revealed the presence of a set of nif genes (nifHDKENB) in strains of Dysgonomonas gadei, Dysgonomonas capnocytophagoides, Saccharicrinis fermentans, and Alkaliflexus imshenetskii. These eight species belonged to and were distributed sporadically within the order Bacteroidales. Acetylene reduction activity was detected in the five species examined, strongly suggesting their diazotrophic nature. Phylogenetic analyses showed monophyletic clustering of the six Nif protein sequences in the eight Bacteroidales species, implying that nitrogen fixation is ancestral to Bacteroidales and has been retained in these species, but lost in many other lineages. The identification of nif genes in Bacteroidales facilitates the prediction of the organismal origins of related sequences directly obtained from various environments.


Illinois medical home project: pilot intervention and evaluation.
A "medical home" provides accessible, continuous, comprehensive, family-centered, coordinated, compassionate, and culturally effective care. This study implemented and evaluated the pilot phase of a pediatric practice-directed medical home quality improvement (QI) project focused on improving primary care for children and youth with special health care needs (CYSHCN). Six practices received training, QI team development and facilitation, and receipt of mini-grants for QI projects. Practice-level and parent-level evaluations were obtained at pre-intervention and post-intervention. The intervention was well-received. Many areas of improvement were found on practice-level evaluations, especially family-centered care and QI activities. Poor response rates limited the interpretation of family-level outcomes; 26% completed baseline outcome evaluations, and 64% of initial respondents completed them post-intervention. Practice education and implementation of QI teams to plan individualized strategies for the practice are feasible ways to seek to improve outcomes for CYSHCN. Modifications of evaluation strategies are needed to enhance response rates for family-level evaluations.


Ca2+ influx and tyrosine kinases trigger Bordetella adenylate cyclase toxin (ACT) endocytosis. Cell physiology and expression of the CD11b/CD18 integrin major determinants of the entry route.
Humans infected with Bordetella pertussis, the whooping cough bacterium, show evidences of impaired host defenses. This pathogenic bacterium produces a unique adenylate cyclase toxin (ACT) which enters human phagocytes and catalyzes the unregulated formation of cAMP, hampering important bactericidal functions of these immune cells that eventually cause cell death by apoptosis and/or necrosis. Additionally, ACT permeabilizes cells through pore formation in the target cell membrane. Recently, we demonstrated that ACT is internalised into macrophages together with other membrane components, such as the integrin CD11b/CD18 (CR3), its receptor in these immune cells, and GM1. The goal of this study was to determine whether ACT uptake is restricted to receptor-bearing macrophages or on the contrary may also take place into cells devoid of receptor and gain more insights on the signalling involved. Here, we show that ACT is rapidly eliminated from the cell membrane of either CR3-positive as negative cells, though through different entry routes, which depends in part, on the target cell physiology and characteristics. ACT-induced Ca(2+) influx and activation of non-receptor Tyr kinases into the target cell appear to be common master denominators in the different endocytic strategies activated by this toxin. Very importantly, we show that, upon incubation with ACT, target cells are capable of repairing the cell membrane, which suggests the mounting of an anti-toxin cell repair-response, very likely involving the toxin elimination from the cell surface.


The genome of the domesticated apple (Malus × domestica Borkh.).
We report a high-quality draft genome sequence of the domesticated apple (Malus × domestica). We show that a relatively recent (>50 million years ago) genome-wide duplication (GWD) has resulted in the transition from nine ancestral chromosomes to 17 chromosomes in the Pyreae. Traces of older GWDs partly support the monophyly of the ancestral paleohexaploidy of eudicots. Phylogenetic reconstruction of Pyreae and the genus Malus, relative to major Rosaceae taxa, identified the progenitor of the cultivated apple as M. sieversii. Expansion of gene families reported to be involved in fruit development may explain formation of the pome, a Pyreae-specific false fruit that develops by proliferation of the basal part of the sepals, the receptacle. In apple, a subclade of MADS-box genes, normally involved in flower and fruit development, is expanded to include 15 members, as are other gene families involved in Rosaceae-specific metabolism, such as transport and assimilation of sorbitol.


Reduction of microalbuminuria by using losartan in normotensive patients with type 2 diabetes mellitus: A randomized controlled trial.
Type 2 diabetes mellitus (T2DM) is a worldwide pandemic that may lead to diabetic kidney disease (DKD), a complication which is the single most important and globally prevalent cause of chronic kidney disease. Microalbuminuria has been shown to be an early indicator of DKD and data suggest that angiotensin receptor blockers (ARBs) reduce urinary albumin excretion and retard the progression of renal disease in hypertensive T2DM patients. However, the effects of ARBs on preventing microalbuminuria and ensuing DKD in normotensive patients with T2DM is yet to be fully established. The objective of this study is to assess the anti-microalbuminuric effects of losar-tan therapy versus placebo in normotensive T2DM patients. This randomized single blinded controlled trial was performed at the Diabetic Clinic, Jinnah Hospital, Lahore over a period of 10 months. A total of 361 normotensive patients with T2DM and microalbuminuria were selected; of them, 171 patients were randomly allocated to the test group and 190 enrolled into the control group. The patients in the test group were started on losartan 50 mg/day for a six month period while those in the control group were put on vitamin B-12 500 mcg/day. The patients as well as the primary attending phy-sicians/lab evaluators were blinded to the study. All study patients were followed up on a monthly basis. Quantitative microalbuminuria was tested at the beginning and at the end of the study. Out of the 171 patients in the test group, 149 (87.1%) had significant reduction of albuminuria by > 30% of their baseline (mean 101.9 +/- 21.7 baseline and, 47.5 +/- 12.9 post-therapy). The corresponding values for albuminuria in the 190 patients in the control group was mean 104.7 +/- 26.3 baseline and post 6-month mean 103.9 +/- 22.9, with P< 0.0001. The anti-albuminuric effect of losartan was reversible as seen on re-checking the urinary albumin two months after discontinuation of treatment. Our study shows that losartan was well tolerated and demonstrated significant anti-proteinuric effects in patients with T2DM with early nephropathy independent of hypertension.


Key principles for the improved conduct of health technology assessments for resource allocation decisions.
Health technology assessment (HTA) is a dynamic, rapidly evolving process, embracing different types of assessments that inform real-world decisions about the value (i.e., benefits, risks, and costs) of new and existing technologies. Historically, most HTA agencies have focused on producing high quality assessment reports that can be used by a range of decision makers. However, increasingly organizations are undertaking or commissioning HTAs to inform a particular resource allocation decision, such as listing a drug on a national or local formulary, defining the range of coverage under insurance plans, or issuing mandatory guidance on the use of health technologies in a particular healthcare system. A set of fifteen principles that can be used in assessing existing or establishing new HTA activities is proposed, providing examples from existing HTA programs. The principal focus is on those HTA activities that are linked to, or include, a particular resource allocation decision. In these HTAs, the consideration of both costs and benefits, in an economic evaluation, is critical. It is also important to consider the link between the HTA and the decision that will follow. The principles are organized into four sections: (i) "Structure" of HTA programs; (ii) "Methods" of HTA; (iii) "Processes for Conduct" of HTA; and (iv) "Use of HTAs in Decision Making."


Comparison of Preoperative Remission Scores and Diabetes Duration Alone as Predictors of Durable Type 2 Diabetes Remission and Risk of Diabetes Complications After Bariatric Surgery: A Post Hoc Analysis of Participants From the Swedish Obese Subjects Study.
Bariatric surgery is associated with diabetes remission and prevention of diabetes-related complications. The ABCD, DiaRem, Ad-DiaRem, DiaBetter, and individualized metabolic surgery scores were developed to predict short- to medium-term diabetes remission after bariatric surgery. However, they have not been tested for predicting durable remission nor the risk of diabetes complications, nor compared with diabetes duration alone. We identified 363 individuals from the surgically treated group in the prospective Swedish Obese Subjects study with preoperative type 2 diabetes and for whom data (preoperative age, BMI, C-peptide, HbA1c, oral diabetes medications, insulin use, and diabetes duration) were available for calculation of remission scores. Partial remission (after 2 and 10 years) was defined as blood glucose <6.1 mmol/L or HbA1c <6.5% (48 mmol/mol) and no diabetes medication. Information on diabetes complications (at baseline and over 15 years of follow-up) was obtained from national health registers. Discrimination was evaluated by area under receiver operating characteristic curves (AUROCs). For 2-year diabetes remission, AUROCs were between 0.79 and 0.88 for remission scores and 0.84 for diabetes duration alone. After 10 years, the predictive ability of scores decreased markedly (AUROCs between 0.70 and 0.76), and no score had higher predictive capacity than diabetes duration alone (AUROC = 0.73). For development of microvascular and macrovascular diabetes complications over 15 years, AUROCs for remission scores were 0.70-0.80 and 0.62-0.71, respectively, and AUROCs for diabetes duration alone were 0.77 and 0.66, respectively. Remission scores and diabetes duration are good predictors of short-term diabetes remission. However, for durable remission and risk of complications, remission scores and diabetes duration alone have limited predictive ability.


Complement component C3 and risk of age-related macular degeneration.
To explore the association between polymorphisms in the complement component 3 (C3) gene and age-related macular degeneration (AMD), and to investigate the modifying effect of complement factor H (CFH) Y402H, LOC387715 A69S and smoking. Pooled data from the prospective, population-based Rotterdam Study (enrolment between 1990 and 1993, and 3 follow-up examinations between September 1, 1993, and December 31, 2004) and an independent case-control study from the Netherlands. The Rotterdam Study comprised a total of 6418 persons aged >or=55 years who had gradable fundus photographs. The case-control study consisted of 357 unrelated AMD patients and 173 control individuals aged >or=55 years. The variants R102G and P314L of the C3 gene, CFH Y402H and LOC387715 A69S, were genotyped in all study participants. Information on cigarette smoking was obtained by interview at baseline. Early and late stages of prevalent and incident AMD, graded according to the international classification and grading system for AMD. We found a population frequency of 0.217 for R102G and 0.211 for P314L in the Rotterdam Study. Both alleles significantly increased the risk of early AMD and all subtypes of late AMD, and this risk seemed to be independent of CFH Y402H, LOC387715 A69S, and smoking. Detailed analysis showed that the haplotype carrying both alleles had the highest frequency difference between cases and controls (P=0.006). We estimated a total population-attributable risk of 14.6%. A meta-analysis of all currently available data yielded a pooled odds ratio (OR) of 1.61 (95% confidence interval [CI], 1.46-1.78) for the R102G allele, and an OR of 1.50 (95% CI, 1.31-1.71) for the P314L allele. Our study showed a significant association between variants in the C3 gene and AMD and further highlights the crucial role of the complement pathway in the etiology of AMD.


A multistage motion vector processing method for motion-compensated frame interpolation.
In this paper, a novel, low-complexity motion vector processing algorithm at the decoder is proposed for motion-compensated frame interpolation or frame rate up-conversion. We address the problems of having broken edges and deformed structures in an interpolated frame by hierarchically refining motion vectors on different block sizes. Our method explicitly considers the reliability of each received motion vector and has the capability of preserving the structure information. This is achieved by analyzing the distribution of residual energies and effectively merging blocks that have unreliable motion vectors. The motion vector reliability information is also used as a prior knowledge in motion vector refinement using a constrained vector median filter to avoid choosing identical unreliable one. We also propose using chrominance information in our method. Experimental results show that the proposed scheme has better visual quality and is also robust, even in video sequences with complex scenes and fast motion.


Genome-wide patterns of transposon proliferation in an evolutionary young hybrid fish.
Hybridization can induce transposons to jump into new genomic positions, which may result in their accumulation across the genome. Alternatively, transposon copy numbers may increase through nonallelic (ectopic) homologous recombination in highly repetitive regions of the genome. The relative contribution of transposition bursts versus recombination-based mechanisms to evolutionary processes remains unclear because studies on transposon dynamics in natural systems are rare. We assessed the genomewide distribution of transposon insertions in a young hybrid lineage ("invasive Cottus", n = 11) and its parental species Cottus rhenanus (n = 17) and Cottus perifretum(n = 9) using a reference genome assembled from long single molecule pacbio reads. An inventory of transposable elements was reconstructed from the same data and annotated. Transposon copy numbers in the hybrid lineage increased in 120 (15.9%) out of 757 transposons studied here. The copy number increased on average by 69% (range: 10%-197%). Given the age of the hybrid lineage, this suggests that they have proliferated within a few hundred generations since admixture began. However, frequency spectra of transposon insertions revealed no increase in novel and rare insertions across assembled parts of the genome. This implies that transposons were added to repetitive regions of the genome that remain difficult to assemble. Future studies will need to evaluate whether recombination-based mechanisms rather than genomewide transposition may explain the majority of the recent transposon proliferation in the hybrid lineage. Irrespectively of the underlying mechanism, the observed overabundance in repetitive parts of the genome suggests that gene-rich regions are unlikely to be directly affected.


The potential protective effect of green, black, red and white tea infusions against adverse effect of cadmium and lead during chronic exposure - A rat model study.
The protective effect of green (GT), black (BT), red (RT) and white (WT) tea infusions on the lungs, brains, hearts, livers and kidneys of adult Wistar rats exposed to Cd (7 mg/kg) and Pb (50 m/kg) was studied. The degree of reduction in the absorption of Cd and Pb in the organs compared to control group and the activity of SOD, CAT and GPx as well as GSH level was evaluated. It was determined that tea significant reduced the accumulation of Cd in the tissues. A significant reduction in the accumulation of Pb was recorded in the brain (WT), liver (GT, WT) and kidneys (BT, GT, RT, WT). A significant increase was observed in the activity of SOD, CAT and GPx in the organs of all rats from tea groups. It was found that the results obtained in rats receiving black, red and white tea were overall not worse than those recorded for rats receiving green tea. The obtained results suggest that drinking tea could be an effective method of reducing the adverse effect of environmental Cd and Pb pollution on the human body.


Respiratory symptoms questionnaire for asthma epidemiology: validity and reproducibility.
There is a need for a new respiratory symptoms questionnaire for use in epidemiological research in asthma. A questionnaire was designed following a pilot study in 78 subjects. It contains nine questions on symptoms such as wheeze and difficulty with breathing in defined circumstances such as exercise and sleep. It was completed by 211 adults and validated by comparison with a self reported history of asthma and with bronchial hyperresponsiveness to histamine. Its short term reproducibility was measured by three repeat administrations over two weeks. Subjects with asthma (n = 33), particularly those having had an asthma attack in the last year (n = 23), were more likely to report any symptom and to report a greater number of symptoms than those without asthma. The same relationship was found for bronchial hyperresponsiveness and symptoms. Either two or more, or three or more, symptoms appeared to be good indices of self reported asthma and bronchial hyperresponsiveness, or both, with a high sensitivity (65-91%) and specificity (85-96%). Reproducibility was good, with few subjects changing the number of symptoms reported by more than one symptom and none by more than four symptoms. The results compared favourably with those from questions on phlegm production from the MRC questionnaire and were better than those reported for the MRC wheeze questions. The questionnaire will be useful for epidemiological research on asthma and could form part of a new standardised questionnaire with wide applications.


Desmocollin 1 expression and desmosomal remodeling during terminal differentiation of human anagen hair follicle: an electron microscopic study.
The terminal differentiation (TD) program of keratinocytes of the human hair follicle (HF) occurs with specific temporal and spatial features in the various layers of the inner root sheath (IRS) and in the innermost layer of the outer root sheath (companion layer). This process is characterized by complex nuclear and cytoplasmic morphological changes, accompanied by profound modifications in intercellular junctions. As no correlation exists between the structure and the molecular composition of desmosomes during TD of the IRS/companion unit, the aim of our study was to investigate by transmission electron microscopy the remodeling of desmosomes in keratinizing cells of these compartments. By immunogold post embedding technique, we studied in anagen HFs the modulation of the synthesis of desmocollin 1 (Dsc1), a transmembrane glycoprotein specifically synthesized in the IRS and in the companion layer. Dsc1 immunoreactivity was actually confined to these compartments and tended to increase just before the level of TD, particularly in the Henle's layer and in the IRS cuticle. In Huxley's layer, the immunolabeling was patchy and in the companion layer Dsc1 synthesis was detected above the level of keratinization of Huxley's layer. In the whole IRS, concomitantly with TD, there was an abrupt and almost complete disappearance of Dsc1 synthesis. An asymmetric distribution of Dsc1 was noticed (i) between cells at different stages of differentiation and (ii) between cells belonging to layers with different spatial/temporal features of TD. Our results show that the ultrastructural modifications of desmosomes during TD of HF are paralleled by the modulation of the synthesis of desmocollin 1.


Intravenously administered vitamin C as cancer therapy: three cases.
Early clinical studies showed that high-dose vitamin C, given by intravenous and oral routes, may improve symptoms and prolong life in patients with terminal cancer. Double-blind placebo-controlled studies of oral vitamin C therapy showed no benefit. Recent evidence shows that oral administration of the maximum tolerated dose of vitamin C (18 g/d) produces peak plasma concentrations of only 220 micromol/L, whereas intravenous administration of the same dose produces plasma concentrations about 25-fold higher. Larger doses (50-100 g) given intravenously may result in plasma concentrations of about 14,000 micromol/L. At concentrations above 1000 micromol/L, vitamin C is toxic to some cancer cells but not to normal cells in vitro. We found 3 well-documented cases of advanced cancers, confirmed by histopathologic review, where patients had unexpectedly long survival times after receiving high-dose intravenous vitamin C therapy. We examined clinical details of each case in accordance with National Cancer Institute (NCI) Best Case Series guidelines. Tumour pathology was verified by pathologists at the NCI who were unaware of diagnosis or treatment. In light of recent clinical pharmacokinetic findings and in vitro evidence of anti-tumour mechanisms, these case reports indicate that the role of high-dose intravenous vitamin C therapy in cancer treatment should be reassessed.


Dietary leucine--an environmental modifier of insulin resistance acting on multiple levels of metabolism.
Environmental factors, such as the macronutrient composition of the diet, can have a profound impact on risk of diabetes and metabolic syndrome. In the present study we demonstrate how a single, simple dietary factor--leucine--can modify insulin resistance by acting on multiple tissues and at multiple levels of metabolism. Mice were placed on a normal or high fat diet (HFD). Dietary leucine was doubled by addition to the drinking water. mRNA, protein and complete metabolomic profiles were assessed in the major insulin sensitive tissues and serum, and correlated with changes in glucose homeostasis and insulin signaling. After 8 weeks on HFD, mice developed obesity, fatty liver, inflammatory changes in adipose tissue and insulin resistance at the level of IRS-1 phosphorylation, as well as alterations in metabolomic profile of amino acid metabolites, TCA cycle intermediates, glucose and cholesterol metabolites, and fatty acids in liver, muscle, fat and serum. Doubling dietary leucine reversed many of the metabolite abnormalities and caused a marked improvement in glucose tolerance and insulin signaling without altering food intake or weight gain. Increased dietary leucine was also associated with a decrease in hepatic steatosis and a decrease in inflammation in adipose tissue. These changes occurred despite an increase in insulin-stimulated phosphorylation of p70S6 kinase indicating enhanced activation of mTOR, a phenomenon normally associated with insulin resistance. These data indicate that modest changes in a single environmental/nutrient factor can modify multiple metabolic and signaling pathways and modify HFD induced metabolic syndrome by acting at a systemic level on multiple tissues. These data also suggest that increasing dietary leucine may provide an adjunct in the management of obesity-related insulin resistance.


Establishing a computed tomography screening clinic.
Since the publication of the National Lung Screening Trial results, there has been increasing interest among radiologists to establish computed tomography (CT) screening centers. During the past 10 years, we have operated a CT screening center in suburban Boston, MA. The purpose of this paper is to describe our process for performing self-referred CT lung screening exams, including our organizational structure, marketing, patient selection process, and standardized workflow. Direct communication between the patient and radiologist, both before and after the CT scan, in our opinion, is critical to a successful screening program. In many ways, CT screening offers radiologists a wonderful opportunity to interact directly with patients. Showing abnormal lung findings to patients on their CT scans may help create a unique "teachable" moment for current smokers and may help motivate patients to quit smoking.


"Did I do as best as the system would let me?" Healthcare professional views on hospital to home care transitions.
Patients are vulnerable to poor quality, fragmented care as they transition from hospital to home. Few studies examine the discharge process from the perspectives of multiple healthcare professionals. To understand care transitions from the perspective of diverse healthcare professionals, and identify recommendations for process improvement. Cross sectional qualitative study. Clinicians, care teams, and administrators from the inpatient general medicine services at one urban, academic hospital; two outpatient primary care clinics; and one Medicaid managed care plan. We conducted 13 focus groups and two in-depth interviews with participants prior to initiating a hospital-funded, multi-component transitional care intervention for uninsured and low-income publicly insured patients, the Care Transitions Innovation (C-TraIn). We used thematic analysis to identify emergent themes and a cross-case comparative analysis to describe variation by participant role and setting. Poor transitional care reflected healthcare system fragmentation, limiting the ability of healthcare professionals to provide optimal patient care. Lack of standardized processes, poor multidisciplinary communication within the hospital, and fragmented communication across settings led to chaotic, unsystematic transitions, poor patient outcomes, and feelings of futility and dissatisfaction among providers. Patients with complex psychosocial needs were especially vulnerable during care transitions. Recommended changes to improve transitional care included improving hospital multidisciplinary hospital rounds, clarifying accountability as patients move across settings, standardizing discharge processes, and providing additional medical staff training. Hospital to home care transitions are critical junctures that can impact health outcomes, experience of care, and costs. Transitional care quality improvement initiatives must address system fragmentation, reduce communication barriers within and between settings, and ensure adequate professional training.


Carbon metabolism and adaptation of hyperalkaliphilic microbes in serpentinizing spring of Manleluag, the Philippines.
Reduced substrates produced by the serpentinization reaction under hydration of olivine may have fuelled biological processes on early Earth. To understand the adaptive strategies and carbon metabolism of the microbes in the serpentinizing ecosystems, we reconstructed 18 draft genomes representing dominant species of Omnitrophicaeota, Gammaproteobacteria and Methanobacteria from the Manleluag serpentinizing spring in Zambales, Philippines (hyperalkaline and rich in methane and hydrogen). Phylogenomics revealed that two genomes were affiliated with a candidate phylum NPL-UPA2 and the references of all our genomes were derived from ground waters, hot springs and the deep biosphere. C1 metabolism appears to be widespread as most of the genomes code for methanogenesis, CO oxidation and CO2 fixation. However, likely due to the low CO2 concentration and election acceptors, the biomass in the spring was extremely low (<103 cell/ml). Various Na+ and K+ transporters and Na+ -driving ATPases appear to be encoded by these genomes, suggesting that nutrient acquisition, bioenergetics and normal cytoplasmic pH were dependent on Na+ and K+ pumps. Our results advance our understanding of the metabolic potentials and bioenergetics of serpentinizing springs and provide a framework of the ecology of early Earth.


Fluoride concentrations in dental plaque and saliva after the use of a fluoride dentifrice preceded by a calcium lactate rinse.
Plaque fluoride concentrations ([F]) are directly related to plaque calcium concentrations [Ca]. Attempts to increase plaque F uptake from dentifrices or rinses have used methods designed to increase plaque [Ca] but with inconsistent results. This double-blind, double-crossover study tested the effect of a 150 mM calcium lactate rinse used prior to brushing with placebo or fluoridated dentifrices (1030 p.p.m. as NaF) on plaque and salivary [F] and [Ca]. Sixteen children (8-10 yr of age) were randomly assigned to four different groups according to the four treatments (placebo dentifrice or fluoridated dentifrice preceded by calcium lactate or deionized water prerinses). Plaque and saliva were collected 1 and 12 h after brushing on day 7 after starting to use the dentifrices. F was determined using the electrode and Ca was determined using atomic absorption spectrometry. Plaque and salivary [Ca] were not significantly increased after use of the calcium lactate prerinse, except for plaque [Ca] 1 h after the use of the placebo dentifrice. A significant increase in salivary [F] was associated with the calcium lactate prerinse only at 1 h after the use of the fluoridated dentifrice. The the calcium lactate prerinse did not significantly affect plaque [F] under any condition.


Diffractive phase elements that synthesize color pseudo-nondiffracting beams.
The design of diffractive phase elements (DPE's) for generating color pseudo-nondiffracting beams (PNDB's) in a multiple-wave illuminating system by the conjugate-gradient method is described. The axial-intensity distributions for dual-color PNDB's generated by the DPE's are shown. The color PNDB's behave as segmented axial-intensity distributions; in each of segments only one color persists. The sequence of wavelength components in the color PNDB's can be arbitrarily preset. Three-dimensional plots of a dual-color PNDB indicate the characteristics of a beam with high transverse resolution.


Modulation of glutathione and antioxidant enzymes by Ocimum sanctum and its role in protection against radiation injury.
Aqueous extract (OE) of the leaves of Ocimum sanctum, the Indian holy basil, has been found to protect mouse against radiation lethality and chromosome damage and to possess significant antioxidant activity in vitro. Therefore a study was conducted to see if OE protects against radiation induced lipid peroxidation in liver and to determine the role, if any, of the inherent antioxidant system in radioprotection by OE. Adult Swiss mice were injected intraperitoneally (i.p.) with 10 mg/kg of OE for 5 consecutive days and exposed to 4.5 Gy of gamma radiation 30 min after the last injection. Glutathione (GSH) and the antioxidant enzymes glutathione transferase (GST), reductase (GSRx), peroxidase (GSPx) and superoxide dismutase (SOD), as well as lipid peroxide (LPx) activity were estimated in the liver at 15 min, 30 min, 1, 2, 4 and 8 hr post-treatment. LPx was also studied after treatment with a single dose of 50 mg/kg of OE with/without irradiation. OE itself increased the GSH and enzymes significantly above normal levels whereas radiation significantly reduced all the values. The maximum decline was at 30-60 min for GSH and related enzymes and at 2 hr for SOD. Pretreatment with the extract checked the radiation induced depletion of GSH and all the enzymes and maintained their levels within or above the control range. Radiation significantly increased the lipid peroxidation rate, reaching a maximum value at 2 hr after exposure (approximately 3.5 times that of control). OE pretreatment significantly (P < 0.0001) reduced the lipid peroxidation and accelerated recovery to normal levels. The results indicate that Ocimum extract protects against radiation induced lipid peroxidation and that GSH and the antioxidant enzymes appear to have an important role in the protection.


The sex-specific association between BMI and coronary heart disease: a systematic review and meta-analysis of 95 cohorts with 1·2 million participants.
The risk of developing coronary heart disease differs by sex, and accumulating evidence suggests that sex differences exist in the effect of coronary risk factors on vascular risk. So far, the existence of a sex difference in the association between BMI and coronary heart disease has not been systematically studied. Since sexual dimorphisms in body composition exist, we postulated that the association between BMI and coronary heart disease would differ between women and men. We did systematic searches of PubMed and Embase up to Feb 20, 2015, for studies of the longitudinal association between BMI and coronary heart disease in women and men from population-based cohorts. We excluded studies if they contained duplicate data from the same study, reported estimates only for Z scores or percentiles of BMI, did not report estimate uncertainty, did not report sex-specific estimates, recruited mainly individuals with a previous history of cardiovascular disease or from within selected populations, and those for which the full text was not available in English. We also included individual participant data from four large studies. Study results were pooled using random-effect models with inverse variance weighting. Our predefined primary endpoint was the pooled women-to-men ratio of the age-adjusted hazard ratios (HRs), or equivalent, relating (continuous and categorical) BMI to coronary heart disease. We reviewed a total of 8561 original entries twice for inclusion in the analysis, of which 32 published studies were eligible for inclusion. Data from 95 cohorts, 1,219,187 participants, and 37,488 incident cases of coronary heart disease were included. Higher BMI was significantly associated with age-adjusted coronary heart disease: for a one-unit (kg/m(2)) increment in BMI; the HR was 1·04 (95% CI 1·03-1·05) in women and 1·05 (1·04-1·07) in men. Compared with people of a normal weight, the age-adjusted HR of coronary heart disease for the underweight group was 1·25 (1·05-1·49) in women and 1·09 (0·91-1·23) in men; for the overweight group 1·20 (1·12-1·29) in women and 1·22 (1·12-1·32) in men; and for the obese group 1·61 (1·42-1·82) in women and 1·60 (1·43-1·79) in men. Overall, these associations did not differ between the sexes. The women-to-men ratios of the HRs were 0·99 (95% CI 0·98-1·00) for a one-unit increment in BMI, 1·10 (0·91-1·32) for the underweight group, 0·99 (0·92-1·07) for the overweight group, and 1·06 (0·95-1·17) for the obese group, relative to the normal weight group. Similar results were obtained after multiple adjustment and in a range of sensitivity analyses. Increased BMI, measured either continuously or categorically, has the same deleterious effects on the risk of coronary heart disease in women and men across diverse populations. None.


Body size and risk of spontaneous abortion among danish pregnancy planners.
Several studies indicate that female obesity increases the risk of spontaneous abortion (SAB). Central adiposity, height, and location of typical weight gain have not been examined as risk factors for SAB. We examined the associations between selected anthropometric factors and risk of SAB among 5132 women enrolled in a Danish Internet-based prospective cohort study of pregnancy planners. We used Cox proportional hazards regression models, with gestational weeks as the time scale, to compute hazard ratios (HRs) of SAB and 95% confidence intervals (CIs). After adjustment for potential confounders, the HRs for SAB among underweight (body mass index (BMI, kg/m(2) ) <20), overweight (BMI: 25-29) and obese (BMI ≥30) women were 1.00 [95% CI: 0.81, 1.24], 0.90 [95% CI: 0.73, 1.09] and 1.23 [95% CI: 0.98, 1.54], respectively, compared with normal weight women (BMI 20-24). The association between obesity and SAB was stronger for early SAB (<8 weeks gestation); HR: 1.34 95% CI: 1.01, 1.77. The HR for height ≥174 cm vs. <166 cm was 0.81 [95% CI: 0.66, 1.00]. Increased waist-to-hip ratio (WHR) was inversely associated with risk of SAB (HR: 0.81; 95% CI: 0.63, 1.05). Waist circumference and location of typical weight gain were not appreciably associated with SAB risk. This study confirms previous studies that have shown a small positive association between obesity and SAB risk. Our results suggest that obesity is a stronger risk factor for early pregnancy losses, and that small stature and low WHR are associated with an increased risk of SAB.


Introduction of oral vitamin D supplementation and the rise of the allergy pandemic.
The history of the allergy pandemic is well documented, enabling us to put the vitamin D hypothesis into its historical context. The purpose of this study is to compare the prevalence of rickets, vitamin D supply, and allergy prevalence at 50-year intervals by means of a retrospective analysis of the literature since 1880. English cities in 1880 were characterized by an extremely high rickets prevalence, the beginning of commercial cod liver oil production, and the near absence of any allergic diseases. By 1930 hay fever prevalence had risen to about 3% in English-speaking countries where cod liver oil was preferentially used for the treatment of rickets. In 1980 vitamin D was used nation-wide in all industrialized countries as supplement to industrial baby food, thus eradicating nearly all cases of rickets. At the same time the allergy prevalence reached an all-time high, affecting about 30% of the population. Time trends are therefore compatible with the vitamin D hypothesis although direct conclusions cannot be drawn. It is interesting, however, to note that there are at least two earlier research papers linking synthesized vitamin D intake and allergy (Reed 1930 and Selye 1962) published prior to the modern vitamin D hypothesis first proposed in 1999.


Transmission of Plasmodium vivax in south-western Uganda: report of three cases in pregnant women.
Plasmodium vivax is considered to be rare in the predominantly Duffy negative populations of Sub-Saharan Africa, as this red blood cell surface antigen is essential for invasion by the parasite. However, despite only very few reports of molecularly confirmed P. vivax from tropical Africa, serological evidence indicated that 13% of the persons sampled in Congo had been exposed to P. vivax. We identified P. vivax by microscopy in 8 smears from Ugandan pregnant women who had been enrolled in a longitudinal study of malaria in pregnancy. A nested polymerase chain reaction (PCR) protocol was used to detect and identify the Plasmodium parasites present. PCR analysis confirmed the presence of P. vivax for three of the women and analysis of all available samples from these women revealed clinically silent chronic low-grade vivax infections for two of them. The parasites in one woman carried pyrimethamine resistance-associated double non-synonymous mutations in the P. vivax dihydrofolate reductase gene. The three women found infected with P. vivax were Duffy positive as were nine of 68 women randomly selected from the cohort. The data presented from these three case reports is consistent with stable transmission of malaria in a predominantly Duffy negative African population. Given the substantial morbidity associated with vivax infection in non-African endemic areas, it will be important to investigate whether the distribution and prevalence of P. vivax have been underestimated in Sub-Saharan Africa. This is particularly important in the context of the drive to eliminate malaria and its morbidity.


Initiation of carbon nanotube growth by well-defined carbon nanorings.
Carbon nanotubes (CNTs), tubular molecular entities that consist of sp(2)-hybridized carbon atoms, are currently produced as mixtures that contain tubes of various diameters and different sidewall structures. The electronic and optical properties of CNTs are determined by their diameters and sidewall structures and so a controlled synthesis of uniform-diameter, single-chirality CNTs-a significant chemical challenge-would provide access to pure samples with predictable properties. Here we report a rational bottom-up approach to synthesize structurally uniform CNTs using carbon nanorings (cycloparaphenylenes) as templates and ethanol as the carbon source. The average diameter of the CNTs formed is close to that of the carbon nanorings used, which supports the operation of a 'growth-from-template' mechanism in CNT formation. This bottom-up organic chemistry approach is intrinsically different from other conventional approaches to making CNTs and, if it can be optimized sufficiently, offers a route to the programmable synthesis of structurally uniform CNTs.


Importance of a three-dimensional measure of humeral head subluxation in osteoarthritic shoulders.
During total shoulder arthroplasty (TSA), humeral head subluxation may be difficult to manage. Furthermore, there is a risk for postoperative recurrence of subluxation, affecting the outcome of TSA. An accurate evaluation of the subluxation is necessary to evaluate this risk. Currently, subluxation is measured in 2 dimensions (2D), usually relative to the glenoid face. The goal of this study was to extend this measure to 3 dimensions (3D) to compare glenohumeral and scapulohumeral subluxation and to evaluate the association of subluxation with the glenoid version. The study analyzed 112 computed tomography scans of osteoarthritic shoulders. We extended the usual 2D definition of glenohumeral subluxation, scapulohumeral subluxation, and glenoid version by measuring their orientation in 3D relative to the scapular plane and the scapular axis. We evaluated statistical associations between subluxation and version in 2D and 3D. Orientation of subluxation and version covered all sectors of the glenoid surface. Scapulohumeral subluxation and glenoid version were highly correlated in amplitude (R(2) = 0.71; P < .01) and in orientation (R(2) = 0.86; P < .01). Approximately every degree of glenoid version induced 1% of scapulohumeral subluxation in the same orientation of the version. Conversely, glenohumeral subluxation was not correlated to glenoid version in 2D or in 3D. Orientation of the humeral subluxation is rarely within the arbitrary computed tomography plane and should therefore be measured in 3D to detect out-of-plane subluxation. Scapulohumeral subluxation and glenoid version measured in 3D could bring valuable information for decision making during TSA.


Development of aquatic toxicity benchmarks for oil products using species sensitivity distributions.
Determining the sensitivity of a diversity of species to spilled oil and chemically dispersed oil continues to be a significant challenge in spill response and impact assessment. We used standardized tests from the literature to develop species sensitivity distributions (SSDs) of acute aquatic toxicity values for several petroleum products and 2 Corexit oil dispersants. Fifth percentile hazard concentrations (HC5s) were computed from the SSDs and used to assess relative oil product toxicity and in evaluating the feasibility of establishing toxicity benchmarks for a community of species. The sensitivity of mysids (Americamysis bahia) and silversides (Menidia beryllina) were evaluated within the SSDs to determine if these common test species were appropriate surrogates for a broader range of species. In general, SSD development was limited by the availability of acute toxicity values that met standardization criteria for a diversity of species. Pooled SSDs were also developed for crude oil and Corexit dispersants because there was only small variability in the HC5s among the individual oil or dispersant products. The sensitivity of mysids and silversides varied across the oil and dispersant products, with the majority of toxicity values greater than the HC5. Application of SSDs appears to be a reasonable approach to developing oil product toxicity benchmarks, but additional toxicity data are needed for a larger range of species conducted under standardized test conditions.


Familial evaluation in arrhythmogenic right ventricular cardiomyopathy: impact of genetics and revised task force criteria.
With recognition of disease-causing genes in arrhythmogenic right ventricular cardiomyopathy, mutation analysis is being applied. The role of genotyping in familial assessment for arrhythmogenic right ventricular cardiomyopathy was investigated, including the prevalence of mutations in known causal genes, the penetrance and expressivity in genotyped families, and the utility of the 2010 Task Force criteria in clinical diagnosis. Clinical and molecular genetic evaluation was performed in 210 first-degree and 45 second-degree relatives from 100 families. In 51 families, the proband was deceased. The living probands had a high prevalence of ECG abnormalities (89%) and ventricular arrhythmia (78%) and evidence of more severe disease than relatives. Definite or probable causal mutations were found in 58% of families and 73% of living probands, of whom 28% had an additional desmosomal variant (ie, mutation or polymorphism). Ninety-three relatives had a causal mutation; 33% fulfilled the 2010 criteria, whereas only 19% satisfied the 1994 version (P=0.03). An additional desmosomal gene variant was found in 10% and was associated with a 5-fold increased risk of developing penetrant disease (odds ratio, 4.7; 95% confidence interval, 1.1 to 20.4; P=0.04). Arrhythmogenic right ventricular cardiomyopathy is a genetically complex disease characterized by marked intrafamilial phenotype diversity. Penetrance is definition dependent and is greater with the 2010 criteria compared with the 1994 criteria. Relatives harboring >1 genetic variant had significantly increased risk of developing clinical disease, potentially an important determinant of the phenotypic heterogeneity seen within families with arrhythmogenic right ventricular cardiomyopathy.