Porphyrins, porphyrin metabolism and porphyrias. I. Update.
The biosynthesis of porphyrins is one of the most conserved parthways known, about the same sequence of reactions taking place in all species. By associating different metals, porphyrins give rise to the "pigments of life": chlorophyll, haem and cobalamin. The unique tetrapyrrolic structure enables it to function in an array of reactions as a single electron carrier and as a catalyst for redox reactions. In this capacity, it constitutes the prosthetic group of enzymes participating in cellular respiration, in conversion reactions involving steroids and lipophilic xenobiotics, in protective mechanisms directed against oxidative stress and in pathways providing central messenger molecules. The formation of haem is accomplished by a sequence of eight dedicated enzymes encoded by different genes, some being active in ubiquitous as well as in erythroid isoforms. Large differences between the participating enzymes with regard to catalytic power, with low capacity steps positioned early in the catalytic chain, constitute a bar against substrate overloading of enzymes processing porphyrins, thus preventing accumulation in the body of these phototoxic compounds under physiological conditions. Most of the haem in the body is produced by the liver and bone marrow, but the mechanisms applied for the control of the synthesis differ between the two organs. The extremely potent hemeprotein enzymes formed in the liver are rapidly turned over in response to current metabolic needs. They have half-lives in the order of minutes or hours and are restored by fast-acting mechanisms for the de novo synthesis, when needed. Uninterrupted and instant availability of the compound is secured by acute deinhibition of the initial enzyme of the synthetic chain, ubiquitous 5-aminolevulinate synthase (ALAS-1), in response to drain of the free cellular haem pool caused by prevailing demands for hemeproteins or by increased catabolism of the compound. In contrast, in the erythroid progenitor cell the haem synthetic machinery is designed for uninterrupted production of huge amounts of haem for combination with globin chains to form hemoglobin at a steady rate. In the erythron the synthesis of the enzymes participating in the formation of haem is under control of erythropoietin, formed under hypoxic conditions. In the absence of iron, to be incorporated in the porphyrin formed in the last step of the synthesis, the mRNA of erythroid 5-aminolevulinate synthase (ALAS-2) is blocked by attachment of an iron-responsive element (IRE) binding cytosolic protein, and transcription of this key enzyme is inhibited. In humans, the genes for each of the haem synthetic enzymes may become the target of mutations that give rise to impaired cellular enzyme activity. Seven of the enzyme deficiencies are associated with accumulation of toxic intermediaries and with disease entities termed porphyrias. The acute porphyrias are characterized by attacks of neuropsychiatric symptoms, which may be due to a toxic surplus of the porphyrin presursor 5-aminolevulinic acid, or a consequence of a deficit of vital hemeproteins resulting from impaired synthesis of haem. In the cutaneous porphyrias, impairment of enzymatic steps where porphyrins are processed gives rise to solar hypersensitivity due to accumulation of phototoxic porphyrins in the skin. Early diagnosis, information to the patient regarding the nature of the illness and counselling aimed at avoidance of triggering factors are cornerstones in the handling of the porphyric diseases. Gene analysis is of incomparable diagnostic reliability in carrier detection, but biochemical methods must be applied in the important task of monitoring porphyric disease activity. In most forms of porphyria the gene carriers run the risk of development of associated diseases in liver or kidneys, a circumstance that prompts application of well-structured surveillance programs.


Transcriptional profiling of nitrogen fixation in Azotobacter vinelandii.
Most biological nitrogen (N(2)) fixation results from the activity of a molybdenum-dependent nitrogenase, a complex iron-sulfur enzyme found associated with a diversity of bacteria and some methanogenic archaea. Azotobacter vinelandii, an obligate aerobe, fixes nitrogen via the oxygen-sensitive Mo nitrogenase but is also able to fix nitrogen through the activities of genetically distinct alternative forms of nitrogenase designated the Vnf and Anf systems when Mo is limiting. The Vnf system appears to replace Mo with V, and the Anf system is thought to contain Fe as the only transition metal within the respective active site metallocofactors. Prior genetic analyses suggest that a number of nif-encoded components are involved in the Vnf and Anf systems. Genome-wide transcription profiling of A. vinelandii cultured under nitrogen-fixing conditions under various metal amendments (e.g., Mo or V) revealed the discrete complement of genes associated with each nitrogenase system and the extent of cross talk between the systems. In addition, changes in transcript levels of genes not directly involved in N(2) fixation provided insight into the integration of central metabolic processes and the oxygen-sensitive process of N(2) fixation in this obligate aerobe. The results underscored significant differences between Mo-dependent and Mo-independent diazotrophic growth that highlight the significant advantages of diazotrophic growth in the presence of Mo.


A near complete, chromosome-scale assembly of the black raspberry (Rubus occidentalis) genome.
The fragmented nature of most draft plant genomes has hindered downstream gene discovery, trait mapping for breeding, and other functional genomics applications. There is a pressing need to improve or finish draft plant genome assemblies. Here, we present a chromosome-scale assembly of the black raspberry genome using single-molecule real-time Pacific Biosciences sequencing and high-throughput chromatin conformation capture (Hi-C) genome scaffolding. The updated V3 assembly has a contig N50 of 5.1 Mb, representing an ∼200-fold improvement over the previous Illumina-based version. Each of the 235 contigs was anchored and oriented into seven chromosomes, correcting several major misassemblies. Black raspberry V3 contains 47 Mb of new sequences including large pericentromeric regions and thousands of previously unannotated protein-coding genes. Among the new genes are hundreds of expanded tandem gene arrays that were collapsed in the Illumina-based assembly. Detailed comparative genomics with the high-quality V4 woodland strawberry genome (Fragaria vesca) revealed near-perfect 1:1 synteny with dramatic divergence in tandem gene array composition. Lineage-specific tandem gene arrays in black raspberry are related to agronomic traits such as disease resistance and secondary metabolite biosynthesis. The improved resolution of tandem gene arrays highlights the need to reassemble these highly complex and biologically important regions in draft plant genomes. The updated, high-quality black raspberry reference genome will be useful for comparative genomics across the horticulturally important Rosaceae family and enable the development of marker assisted breeding in Rubus.


Discovery of WD Repeat-Containing Protein 5 (WDR5)-MYC Inhibitors Using Fragment-Based Methods and Structure-Based Design.
The frequent deregulation of MYC and its elevated expression via multiple mechanisms drives cells to a tumorigenic state. Indeed, MYC is overexpressed in up to ∼50% of human cancers and is considered a highly validated anticancer target. Recently, we discovered that WD repeat-containing protein 5 (WDR5) binds to MYC and is a critical cofactor required for the recruitment of MYC to its target genes and reported the first small molecule inhibitors of the WDR5-MYC interaction using structure-based design. These compounds display high binding affinity, but have poor physicochemical properties and are hence not suitable for in vivo studies. Herein, we conducted an NMR-based fragment screening to identify additional chemical matter and, using a structure-based approach, we merged a fragment hit with the previously reported sulfonamide series. Compounds in this series can disrupt the WDR5-MYC interaction in cells, and as a consequence, we observed a reduction of MYC localization to chromatin.


Exogenous glucose-dependent insulinotropic polypeptide worsens post prandial hyperglycemia in type 2 diabetes.
Glucose-dependent insulinotropic polypeptide (GIP), unlike glucagon-like peptide (GLP)-1, lacks glucose-lowering properties in patients with type 2 diabetes. We designed this study to elucidate the underlying pathophysiology. Twenty-two insulin-naïve subjects with type 2 diabetes were given either synthetic human GIP (20 ng x kg(-1) x min(-1)) or placebo (normal saline) over 180 min, starting with the first bite of a mixed meal (plus 1 g of acetaminophen) on two separate occasions. Frequent blood samples were obtained over 6 h to determine plasma GIP, GLP-1, glucose, insulin, glucagon, resistin, and acetaminophen levels. Compared with placebo, GIP induced an early postprandial increase in insulin levels. Intriguingly, GIP also induced an early postprandial augmentation in glucagon, a significant elevation in late postprandial glucose, and a decrease in late postprandial GLP-1 levels. Resistin and acetaminophen levels were comparable in both interventions. By immunocytochemistry, GIP receptors were present on human and mouse alpha-cells. In alphaTC1 cell line, GIP induced an increase in intracellular cAMP and glucagon secretion. CONCLUSIONS; GIP, given to achieve supraphysiological plasma levels, still had an early, short-lived insulinotropic effect in type 2 diabetes. However, with a concomitant increase in glucagon, the glucose-lowering effect was lost. GIP infusion further worsened hyperglycemia postprandially, most likely through its suppressive effect on GLP-1. These findings make it unlikely that GIP or GIP receptor agonists will be useful in treating the hyperglycemia of patients with type 2 diabetes.


Prospective external validation of the Predicting Out-of-OFfice Blood Pressure (PROOF-BP) strategy for triaging ambulatory monitoring in the diagnosis and management of hypertension: observational cohort study.
To prospectively validate the Predicting Out-of-OFfice Blood Pressure (PROOF-BP) algorithm to triage patients with suspected high blood pressure for ambulatory blood pressure monitoring (ABPM) in routine clinical practice. Prospective observational cohort study. 10 primary care practices and one hospital in the UK. 887 consecutive patients aged 18 years or more referred for ABPM in routine clinical practice. All underwent ABPM and had the PROOF-BP applied. The main outcome was the proportion of participants whose hypertensive status was correctly classified using the triaging strategy compared with the reference standard of daytime ABPM. Secondary outcomes were the sensitivity, specificity, and area under the receiver operator characteristic curve (AUROC) for detecting hypertension. The mean age of participants was 52.8 (16.2) years. The triaging strategy correctly classified hypertensive status in 801 of the 887 participants (90%, 95% confidence interval 88% to 92%) and had a sensitivity of 97% (95% confidence interval 96% to 98%) and specificity of 76% (95% confidence interval 71% to 81%) for hypertension. The AUROC was 0.86 (95% confidence interval 0.84 to 0.89). Use of triaging, rather than uniform referral for ABPM in routine practice, would have resulted in 435 patients (49%, 46% to 52%) being referred for ABPM and the remainder managed on the basis of their clinic measurements. Of these, 69 (8%, 6% to 10%) would have received treatment deemed unnecessary had they received ABPM. In a population of patients referred for ABPM, this new triaging approach accurately classified hypertensive status for most, with half the utilisation of ABPM compared with usual care. This triaging strategy can therefore be recommended for diagnosis or management of hypertension in patients where ABPM is being considered, particularly in settings with limited resources.


Anatomical considerations of the internal iliac artery in association with the ilioinguinal approach for anterior acetabular fracture fixation.
Vascular injury may be encountered during an anterior approach to the pelvis or acetabulum-be it due to hematoma decompression, clot dislodgement during fracture manipulation, or iatrogenic. This can be associated with significant bleeding, hemodynamic instability, and subsequent morbidity. If the exact source of bleeding cannot be easily identified, compression of the internal iliac artery may be a lifesaving procedure. We describe an extension of the lateral window of the ilioinguinal (or Olerud) approach elaborated on cadavers. The approach allows emergent access the internal iliac artery and intraoperative cross-clamping of the internal iliac vessels to control bleeding. The approach allows rapid access to the internal iliac artery. The surgeon should be familiar, however, with the surgical anatomy of this region to avoid potential injury to the ureter, peritoneum, lymphatics, and sympathetic nerves overlying the vessels when using the approach described.


Minoxidil upregulates the expression of vascular endothelial growth factor in human hair dermal papilla cells.
The hair follicle dermal papilla which controls hair growth, is characterized in the anagen phase by a highly developed vascular network. We have demonstrated in a previous study that the expression of an angiogenic growth factor called vascular endothelial growth factor (VEGF) mRNA varied during the hair cycle. VEGF mRNA is strongly expressed in dermal papilla cells (DPC) in the anagen phase, but during the catagen and telogen phases. VEGF mRNA is less strongly expressed. This involvement of VEGF during the hair cycle allowed us to determine whether VEGF mRNA expression by DPC was regulated by minoxidil. In addition, the effect of minoxidil on VEGF protein synthesis in both cell extracts and DPC-conditioned medium, was investigated immunoenzymatically. Both VEGF mRNA and protein were significantly elevated in treated DPC compared with controls. DPC incubated with increasing minoxidil concentrations (0.2, 2, 6, 12 and 24 mumol/L) induced a dose-dependent expression of VEGF mRNA. Quantification of transcripts showed that DPC stimulated with 24 mumol/L minoxidil express six times more VEGF mRNA than controls. Similarly, VEGF protein production increases in cell extracts and conditioned media following minoxidil stimulation. These studies strongly support the likely involvement of minoxidil in the development of dermal papilla vascularization via a stimulation of VEGF expression, and support the hypothesis that minoxidil has a physiological role in maintaining a good vascularization of hair follicles in androgenetic alopecia.


Improving malaria treatment and prevention in India by aiding district managers to manage their programmes with local information: a trial assessing the impact of Lot Quality Assurance Sampling on programme outcomes.
This paper reports the first trial of Lot Quality Assurance Sampling (LQAS) assessing associations between access to LQAS data and subsequent improvements in district programming. This trial concerns India's approach to addressing an increase in malaria-attributable deaths by training community health workers to diagnose, treat and prevent malaria, while using LQAS to monitor sub-district performance and make programme improvements. The Ministry of Health introduced LQAS into four matched high malaria burden districts (Annual Parasite Incidence >5) (N > 5 million). In each sub-district, we sampled four populations in three 6-monthly surveys: households, children <5 years, people with fever in the last 2 weeks and community health workers. In three districts, trained local staff collected, analysed and used data for programme management; in one control district, non-local staff collected data and did not disseminate results. For eight indicators, we calculated the change in proportion from survey one to three and used a Difference-in-Differences test to compare the relative change between intervention and control districts. Coverage increased from survey one to three for 24 of 32 comparisons. Difference-in-Differences tests revealed that intervention districts exhibited significantly greater change in four of six vertical strategies (insecticide treated bed-nets and indoor residual spraying), one of six treatment-seeking behaviours and four of 12 health worker capacity indicators. The control district displayed greater improvement than two intervention districts for one health worker capacity indicator. One district with poor management did not improve. In this study, LQAS results appeared to support district managers to increase coverage in underperforming areas, especially for vertical strategies in the presence of diligent managers.


Selenium supplementation inhibits IGF-1 signaling and confers methionine restriction-like healthspan benefits to mice.
Methionine restriction (MR) dramatically extends the healthspan of several organisms. Methionine-restricted rodents have less age-related pathology and increased longevity as compared with controls, and recent studies suggest that humans might benefit similarly. Mechanistically, it is likely that the decreased IGF-1 signaling that results from MR underlies the benefits of this regimen. Thus, we hypothesized that interventions that decrease IGF-1 signaling would also produce MR-like healthspan benefits. Selenium supplementation inhibits IGF-1 signaling in rats and has been studied for its putative healthspan benefits. Indeed, we show that feeding mice a diet supplemented with sodium selenite results in an MR-like phenotype, marked by protection against diet-induced obesity, as well as altered plasma levels of IGF-1, FGF-21, adiponectin, and leptin. Selenomethionine supplementation results in a similar, albeit less robust response, and also extends budding yeast lifespan. Our results indicate that selenium supplementation is sufficient to produce MR-like healthspan benefits for yeast and mammals.


A New Normal or Business-as-Usual? Lessons for COVID-19 from Financial Crises in East and Southeast Asia.
Crises and dislocations home in on social, economic, and political weaknesses that are often sidestepped or pushed to the backburner in the interests of master plans of growth or development. Recovery from crises, then, provides the opportunity to address these underlying issues that preceded and, likely, contributed to the crises or dislocation; meanwhile, a return to the previous normalcy following such crises generally means exacerbation of these weaknesses that erode and threaten to fracture social, economic and political foundations. This paper documents social and economic policies across two financial crises, the Asian Financial Crisis and the Global Financial Crisis, for South Korea, the Philippines, and Indonesia, to reveal the problems from growth-centric recovery focus on economic fragilities, social cohesion, and political stability. Further, using evidence from the ground and survey data, we also show how recovery to a new normal with a reprioritization of social policies invigorates the social, political, and economic foundations. We round off the study with an examination of social policy changes under COVID-19 to assess how the efforts track against a recovery to business-as-usual economic normalcy or a new normal that reprioritizes social policies and the economy. The scope of change is high; as we show in the paper, it is also necessary.


Safety of oral ivermectin during pregnancy: a systematic review and meta-analysis.
About 3·7 billion doses of ivermectin have been distributed in mass drug administration (MDA) campaigns globally over the past 30 years. At 10-100 times higher than current human doses, ivermectin is a known teratogen in mammals. During these campaigns with recommended doses, pregnant women might be inadvertently exposed. We therefore aimed to evaluate the existing evidence for serious and non-serious adverse events after ivermectin exposure in pregnant women. For this systematic review and meta-analysis, we searched relevant databases and trial registry platforms on July 15, 2018, for randomised controlled trials (RCTs) and observational studies that reported adverse events in pregnant women. We did not use language or date restrictions. Outcomes of interest were spontaneous abortions, stillbirths, congenital anomalies, and neonatal death (serious adverse events), as well as maternal morbidity, preterm births, and low birthweight (adverse events). The risk of bias was assessed using the Newcastle-Ottawa Scale for observational studies and the Cochrane Risk of Bias Tool for RCTs. We did the meta-analysis of observational studies and RCTs separately. The quality of evidence was assessed using the GRADE approach. The study protocol is registered with PROSPERO, protocol CRD42016046914. We identified 147 records, of which only five observational studies and one RCT were included for quantitative analysis; these studies were published between 1990 and 2008, and were done in six African countries. 893 women with 899 pregancy outcomes were included, of whom 496 pregnant women (500 pregnancy outcomes) received ivermectin inadvertently during MDA campaigns in the observational studies and 397 pregnant women (399 pregnancy outcomes) purposely received ivermectin as part of the open-label RCT. No study reported neonatal deaths, maternal morbidity, preterm births, or low birthweight. It is unclear whether exposure to ivermectin during pregnancy increases the risk of spontaneous abortions and stillbirths (odds ratio [OR] 1·15 [95% CI 0·75-1·78] with very low certainty of evidence for the four observational studies and 0·62 [0·18-2·14] with very low certainty of evidence for the RCT) or congenital anomalies (OR 1·69 [95% CI 0·83-3·41] with very low certainty of evidence for the five observational studies and 1·10 [0·07-17·65] with very low certainty of evidence for the RCT). There is insufficient evidence to conclude on the safety profile of ivermectin during pregnancy. Treatment campaigns should focus additional efforts on preventing inadvertent treatment of pregnant women. Unitaid.


Energetic and ecological constraints on population density of reef fishes.
Population ecology has classically focused on pairwise species interactions, hindering the description of general patterns and processes of population abundance at large spatial scales. Here we use the metabolic theory of ecology as a framework to formulate and test a model that yields predictions linking population density to the physiological constraints of body size and temperature on individual metabolism, and the ecological constraints of trophic structure and species richness on energy partitioning among species. Our model was tested by applying Bayesian quantile regression to a comprehensive reef-fish community database, from which we extracted density data for 5609 populations spread across 49 sites around the world. Our results indicate that population density declines markedly with increases in community species richness and that, after accounting for richness, energetic constraints are manifested most strongly for the most abundant species, which generally are of small body size and occupy lower trophic groups. Overall, our findings suggest that, at the global scale, factors associated with community species richness are the major drivers of variation in population density. Given that populations of species-rich tropical systems exhibit markedly lower maximum densities, they may be particularly susceptible to stochastic extinction.


Effect of hyperbaric oxygenation on normal and chronic streptozotocin diabetic peripheral nerves.
Hyperbaric oxygenation is known to affect energy metabolism and endothelial cell structure and function, but its effects on peripheral nerve have not been reported. We investigated whether it would (i) reverse established streptozotocin-induced diabetic neuropathy, a condition in which endoneurial hypoxia exists; (ii) affect energy metabolism in nerve; and (iii) alter the blood-nerve barrier. Sprague-Dawley rats that had been diabetic for 3 months and age-matched controls were used in these studies. One diabetic group and one control group were treated with hyperbaric oxygenation (2 atm for 2 h, 5 days/week) for 4 weeks. Identical groups remained in room air. Sciatic nerve adenosine triphosphate (ATP), creatine phosphate, lactate, and glucose concentrations showed similar changes at rest in both room air and after hyperbaric oxygenation. Nerves of control and diabetic groups exhibited increased lactate production and increased utilization of glucose, ATP, and creatine phosphate after 15 min of anoxia. The albumin blood-nerve barrier index was increased in control and diabetic nerves after hyperbaric treatment. Nerve conduction velocity was reduced in the diabetic-room air group and not improved by hyperbaric oxygenation. Caudal nerve action potential, which was significantly reduced in this group, was normalized after hyperbaric treatment. Resistance to ischemic conduction failure was increased in untreated diabetic nerve but not significantly different from controls after hyperbaric exposure. These findings indicate that treatment with hyperbaric oxygenation will partially reverse the neuropathy encountered in chronic diabetes. The biochemical changes are suggestive of enhanced nerve energy metabolism induced by hyperbaric oxygenation. The altered albumin blood-nerve barrier index presumably results from the action of free radicals on endothelial cells.


Pro-inflammatory cytokines attenuate glucose-stimulated insulin secretion from INS-1E insulinoma cells by restricting mitochondrial pyruvate oxidation capacity - Novel mechanistic insight from real-time analysis of oxidative phosphorylation.
Pro-inflammatory cytokines cause pancreatic beta cell failure during the development of type 2 diabetes. This beta cell failure associates with mitochondrial dysfunction, but the precise effects of cytokines on mitochondrial respiration remain unclear. To test the hypothesis that pro-inflammatory cytokines impair glucose-stimulated insulin secretion (GSIS) by inhibiting oxidative ATP synthesis, we probed insulin release and real-time mitochondrial respiration in rat INS-1E insulinoma cells that were exposed to a combination of 2 ng/mL interleukin-1-beta and 50 ng/mL interferon-gamma. We show that 24-h exposure to these cytokines dampens both glucose- and pyruvate-stimulated insulin secretion (P < 0.0001 and P < 0.05, respectively), but does not affect KCl-induced insulin release. Mirroring secretory defects, glucose- and pyruvate-stimulated mitochondrial respiration are lowered after cytokine exposure (P < 0.01). Further analysis confirms that cytokine-induced mitochondrial respiratory defects occur irrespective of whether fuel oxidation is coupled to, or uncoupled from, ATP synthesis. These observations demonstrate that pro-inflammatory cytokines attenuate GSIS by restricting mitochondrial pyruvate oxidation capacity. Interleukin-1-beta and interferon-gamma also increase mitochondrial superoxide levels (P < 0.05), which may reinforce the inhibition of pyruvate oxidation, and cause a modest (20%) but significant (P < 0.01) loss of INS-1E cells. Cytokine-induced INS-1E cell failure is insensitive to palmitoleate and linoleate, which is at odds with the cytoprotection offered by unsaturated fatty acids against harm caused by nutrient excess. Our data disclose a mitochondrial mechanism for cytokine-impaired GSIS in INS-1E cells, and suggest that inflammatory and nutrient-related beta cell failure emerge, at least partly, through distinct paths.


Description of a female and variation of Microgomphus alani Kosterin, 2016 (Odonata: Gomphidae) in Cambodia, with a note on sexual dimorphism in Microgomphus spp.
Microgomphus alani Kosterin, 2016 was described from two teneral males from the Cardamom Mts. in SW Cambodia; it turned out that the description was based on the holotype with the anal appendages not fully expanded. Further specimens of M. alani (all mature) are reported: a male and two females from the Phnom Kulen Mts, NW Cambodia and five males and one female from Mondulkiri Province, E Cambodia. The appendage shape in mature males is illustrated and discussed and the female is described for the first time. The specimens from E Cambodia are ca 15-20% larger than those from SW and NW Cambodia and exhibit some differences in the shapes of the male epiproct and anterior hamulus and face maculation, however evaluated as insufficient to erect a new taxon. A hitherto neglected manifestation of sexual dimorphism in probably most Microgomphus spp. and at least some Heliogomphus spp. is pointed out. It concerns the mid- and metafemur, in males armed with numerous, dense and small spinulets but with two rows of long, sparse spines in females. An earlier report of Microgomphus jurzitzai Karube, 2000 for Cambodia was based on a misidentified M. alani specimen and is cancelled. A female specimen earlier identified and described from Thailand as Heliogomphus selysi Fraser, 1925 most probably was M. alani as well.


Uneven distribution of postjunctional alpha 1-and alpha 2-like adrenoceptors in canine arterial and venous smooth muscle.
We studied isolated canine arteries and veins to compare the pharmacological properties of their postjunctional alpha-adrenoceptors. Rings of femoral and splenic arteries and of femoral and saphenous veins were mounted for isometric tension recording in organ chambers filled with Krebs-Ringer bicarbonate solution. The four blood vessels contracted when exposed to methoxamine, norepinephrine, phenylephrine, and tramazoline; clonidine failed to induce contraction only in the splenic artery. The relative sensitivity to methoxamine was comparable in the arteries and veins, but that for phenylephrine was larger in the former. The veins, but not the arteries, were more sensitive to clonidine and tramazoline than to phenylephrine or methoxamine. Phentolamine was a competitive antagonist against norepinephrine in the arteries and the veins. Prazosin was a competitive antagonist arteries. The competitive antagonistic properties of yohimbine were more pronounced in the veins than in the arteries. Verapamil depressed to the same extent the contractile responses of saphenous veins to clonidine and norepinephrine, but reduced the contractions caused by methoxamine more than those due to norepinephrine. These results indicate the presence of both alpha 1- and alpha 2-like adrenoceptors on venous smooth muscle cells, whereas arterial smooth muscle cells contain mainly postjunctional alpha 1-adrenoceptors.


Self-reported noise exposure as a risk factor for long-term sickness absence.
Self-reported noise exposure is on the rise in Denmark. Little is known, however, about the social consequences, including sickness absence, of noise exposure. The aim of this paper was to investigate the association between self-reported noise exposure and long-term sickness absence. The association was investigated using the Cox proportional hazards model to analyze outcomes in Danish register data on the basis of Danish survey data (5357 employees aged 18-69 in 2000). The analyses showed that self-reported noise exposure was significantly associated with long-term sickness absence for both men and women when adjusting for demographic factors and health behavior. After further adjustment for physical workload at work the association between noise exposure and sickness absence disappeared for women, but not for men. Men that reported to be exposed to loud noise between one-quarter and three-quarters of their time at work had an increased risk of 43% (CI: 10-85%) for sickness absence of two weeks or longer compared to men that reported never to be exposed to loud noise. Men that reported to be little/rarely exposed to loud noise had an increased risk of 37% (CI: 7-76%). Men that reported to be exposed to loud noise more than three-quarters of their time at work did not have an increased risk of sickness absence. This latter result might be due a healthy worker effect and/or more frequent use of hearing protection in this group. Along with evidence from previous studies these results provide further support for an association between occupational noise exposure and sickness absence.


Exact Distribution of Linkage Disequilibrium in the Presence of Mutation, Selection, or Minor Allele Frequency Filtering.
Linkage disequilibrium (LD), often expressed in terms of the squared correlation (r 2) between allelic values at two loci, is an important concept in many branches of genetics and genomics. Genetic drift and recombination have opposite effects on LD, and thus r 2 will keep changing until the effects of these two forces are counterbalanced. Several approximations have been used to determine the expected value of r 2 at equilibrium in the presence or absence of mutation. In this paper, we propose a probability-based approach to compute the exact distribution of allele frequencies at two loci in a finite population at any generation t conditional on the distribution at generation t - 1. As r 2 is a function of this distribution of allele frequencies, this approach can be used to examine the distribution of r 2 over generations as it approaches equilibrium. The exact distribution of LD from our method is used to describe, quantify, and compare LD at different equilibria, including equilibrium in the absence or presence of mutation, selection, and filtering by minor allele frequency. We also propose a deterministic formula for expected LD in the presence of mutation at equilibrium based on the exact distribution of LD.


T1 mapping in patients with acute myocardial infarction.
Pixel-by-pixel calculation of T1 values (T1 mapping) has been used in different tissues to focus on T1 changes in a quantitative fashion. The aim of this study was to establish T1 mapping of human myocardium on a 1.5 Tesla system and to examine its diagnostic potential in patients with acute myocardial infarction (AMI). 8 patients with reperfused AMI (day 3 +/- 1) underwent multi-breath-hold MRI in a 1.5 Tesla system. Sets of five images with varying T1 weighting were acquired prior to and after the administration of contrast agent to generate images from calculated T1 values (T1 mapping). Prior to the contrast agent administration, all patients showed T1 prolongation in the area of infarction, which was identified in separate measurements using the delayed enhancement approach. Compared to noninfarcted areas, T1 values in the infarcted areas were increased by 18 +/- 7% (SE, p < 0.05). The spatial extent of the area of T1 prolongation was larger than that of the hyper-enhanced areas in conventional contrast-enhanced images. T1 maps obtained after the application of Gadolinium-DTPA revealed a T1 reduction of 27 +/- 4% in infarcted tissue compared to noninfarcted areas (p < 0.05). The areas showing T1 reduction were in agreement with the hyper-enhanced regions in conventional T1-weighted images. T1 mapping visualizes changes in the longitudinal relaxation time induced by AMI. T1 mapping can detect myocardial necrosis without the use of contrast media. Information that can be extracted from a combination of pre- and postcontrast T1 maps exceeds that from conventional contrast studies.


Dietary glycemic load and cancer recurrence and survival in patients with stage III colon cancer: findings from CALGB 89803.
The influence of glycemic load and related measures on survival among colon cancer patients remains largely unknown. We conducted a prospective, observational study of 1011 stage III colon cancer patients reporting dietary intake during and 6 months after participation in an adjuvant chemotherapy trial. We examined the influence of glycemic load, glycemic index, fructose, and carbohydrate intakes on cancer recurrence and mortality using Cox proportional hazards regression; all tests of statistical significance were two-sided. Stage III colon cancer patients in the highest quintile of dietary glycemic load experienced an adjusted hazard ratio (HR) for disease-free survival of 1.79 (95% confidence interval [CI] = 1.29 to 2.48), compared with those in the lowest quintile (P (trend) across quintiles <.001). Increased glycemic load was associated with similar detriments in recurrence-free (P (trend) across quintiles <.001) and overall survival (P (trend) across quintiles <.001). These associations differed statistically significant by body mass index (BMI) (P (interaction) =.01). Whereas glycemic load was not associated with disease-free survival in patients with BMI < 25kg/m(2), higher glycemic load was statistically significant associated with worse disease-free survival among overweight or obese participants (BMI ≥ 25kg/m(2); HR = 2.26; 95% CI = 1.53 to 3.32; P (trend) across quintiles <.001). Increasing total carbohydrate intake was similarly associated with inferior disease-free, recurrence-free, and overall survival (P (trend) across quintiles <.001). Higher dietary glycemic load and total carbohydrate intake were statistically significant associated with an increased risk of recurrence and mortality in stage III colon cancer patients. These findings support the role of energy balance factors in colon cancer progression and may offer potential opportunities to improve patient survival.


Solitary superficial microcystic lymphatic malformation (lymphangioma circumscriptum) of the gingiva.
Lymphatic malformations are uncommon, hamartomatous, developmental aberrations of the lymphatic system. The case presented in this report is a rare solitary gingival involvement of a microcystic-type lymphatic malformation. The lesion presented clinically as a small vesicular swelling of a buccal interdental papilla in a 16-year-old girl. Involved tissues were excised and submitted for routine histologic examination. The expression of the endothelial marker CD34 was investigated using immunohistochemical staining. A physical examination failed to reveal similar or other abnormalities elsewhere in the body of the patient, including the oral cavity. Histopathologic analysis of the specimen demonstrated the presence of subepithelial, thin-walled, distended vascular cavities forming confluent vesicles containing lymph. The dilated lymphatic formations were lined by flattened CD34-negative endothelial cells. These features were consistent with a microcystic gingival lymphatic malformation. To the best of our knowledge, only two additional reports of this malformation have been published to date, but both presented with bilateral gingival involvement. Even though lymphatic malformations are encountered very infrequently on gingiva, they should be considered in the differential diagnosis of related conditions with a vesicular clinical appearance.


Acute effects of night work and meals on blood glucose levels.
Employees working at night are at increased risk of diabetes. A possible mechanism is related to differences in glucose regulation at night. Laboratory simulated night work studies show regulation of blood glucose is impaired at night. Regular exposure to high glucose levels at night may explain the observed relationship between night work and diabetes. We performed a field study of 19 nonsmoking women from the health-care sector to investigate how night work and the composition of meals affect post-prandial blood glucose levels. Blood glucose levels were self-assessed by finger-prick blood sampling using the Beurer blood glucose monitoring system. Measurements were done before and 15, 30, 60, and 120 min after different test meals: a nighttime high sugar meal during a night shift and during a day shift, and a reference (low sugar) meal under these same two conditions. There was a statistically significant difference in blood glucose concentration between the four test meal conditions (P = .0086). Post-meal blood glucose levels following the night-shift meals, compared to following daytime meals, rose faster and remained elevated for longer a duration of time. At the 15 min time point following the high sugar test meal, the blood glucose concentration was 8.3 mmol/L when consumed at night vs. 7.3 mmol/L when consumed during the day. We found no difference in area under the blood glucose concentration-time curve (AUC) after consumption of the high or low sugar test meals during the night shift compared with consumption of them during the day. Our findings indicate the glucose levels in response to food intake by female night working healthcare assistants are higher following the nighttime compared with daytime consumption of a high sugar content meal. However, we did not find a difference in total glucose exposure across time (assessed as AUC) after eating a high vs. low sugar meal during the night shift.


Identification, cloning and expression of the mouse N-acetylglutamate synthase gene.
In ureotelic animals, N-acetylglutamate (NAG) is an essential allosteric activator of carbamylphosphate synthetase I (CPSI), the first enzyme in the urea cycle. NAG synthase (NAGS; EC 18.104.22.168) catalyses the formation of NAG from glutamate and acetyl-CoA in liver and intestinal mitochondria. This enzyme is supposed to regulate ureagenesis by producing variable amounts of NAG, thus modulating CPSI activity. Moreover, inherited deficiencies in NAGS have been associated with hyperammonaemia, probably due to the loss of CPSI activity. Although the existence of the NAGS protein in mammals has been known for decades, the gene has remained elusive. We identified the mouse (Mus musculus) and human NAGS genes using their similarity to the respective Neurospora crassa gene. NAGS was cloned from a mouse liver cDNA library and was found to encode a 2.3 kb message, highly expressed in liver and small intestine with lower expression levels in kidney, spleen and testis. The deduced amino acid sequence contains a putative mitochondrial targeting signal at the N-terminus. The cDNA sequence complements an argA (NAGS)-deficient Escherichia coli strain, reversing its arginine auxotrophy. His-tagged versions of the pre-protein and two putative mature proteins were each overexpressed in E. coli, and purified to apparent homogeneity by using a nickel-affinity column. The pre-protein and the two putative mature proteins catalysed the NAGS reaction but one of the putative mature enzymes had significantly higher activity than the pre-protein. The addition of l-arginine increased the catalytic activity of the purified recombinant NAGS enzymes by approx. 2-6-fold.


Cationic lipid-assisted polymeric nanoparticle mediated GATA2 siRNA delivery for synthetic lethal therapy of KRAS mutant non-small-cell lung carcinoma.
Synthetic lethal interaction provides a conceptual framework for the development of wiser cancer therapeutics. In this study, we exploited a therapeutic strategy based on the interaction between GATA binding protein 2 (GATA2) downregulation and the KRAS mutation status by delivering small interfering RNA targeting GATA2 (siGATA2) with cationic lipid-assisted polymeric nanoparticles for treatment of non-small-cell lung carcinoma (NSCLC) harboring oncogenic KRAS mutations. Nanoparticles carrying siGATA2 (NPsiGATA2) were effectively taken up by NSCLC cells and resulted in targeted gene suppression. NPsiGATA2 selectively inhibited cell proliferation and induced cell apoptosis in KRAS mutant NSCLC cells. However, this intervention was harmless to normal KRAS wild-type NSCLC cells and HL7702 hepatocytes, confirming the advantage of synthetic lethality-based therapy. Moreover, systemic delivery of NPsiGATA2 significantly inhibited tumor growth in the KRAS mutant A549 NSCLC xenograft murine model, suggesting the therapeutic promise of NPsiGATA2 delivery in KRAS mutant NSCLC therapy.


Performance improvement CME: schizoaffective disorder.
Correctly diagnosing schizoaffective disorder depends on having an accurate history of the patient's signs and symptoms, including documentation from medical and psychiatric records and information gathered from patients and their family members or caregivers. Placing the patient's symptoms within a diagnostic framework helps to differentiate schizoaffective disorder from other psychotic disorders so that an individualized, evidence-based treatment plan can be formulated. Clinicians should monitor treatment adherence, which is closely correlated with treatment outcomes; using multiple, objective measures to assess adherence is optimal. This performance improvement activity provides opportunities to improve strategies for diagnosing schizoaffective disorder, individualizing treatment, and assessing and managing medication nonadherence.


Dissipative soliton resonance in a passively mode-locked fiber laser.
The phenomenon of dissipative soliton resonance (DSR) predicts that an increase of pulse energy by orders of magnitude can be obtained in laser oscillators. Here, we prove that DSR is achievable in a realistic ring laser cavity using nonlinear polarization evolution as the mode-locking mechanism, whose nonlinear transmission function is adjusted through a set of waveplates and a passive polarizer. The governing model accounts explicitly for the arbitrary orientations of the waveplates and the polarizer, as well as the gain saturation in the amplifying medium. It is shown that DSR is achievable with realistic laser settings. Our findings provide an excellent design tool for optimizing the mode-locking performance and the enhancement of energy delivered per pulse by orders of magnitude.


An atypical case of vasospastic angina: demonstrating the usefulness of Holter monitoring.
Prinzmetal variant angina or vasospastic angina is a clinical syndrome characterised by episodic chest pain and transient reversible ST segment changes in ECG. The aetiology and treatment of the condition are quite different compared to those of conventional coronary artery disease. The crux of the problem in variant angina patients remains the diagnosis, especially if coexisting coronary artery disease is present. Timely diagnosis not only helps relieve symptoms, it also prevents complications such as ventricular tachycardia. Conventional provocative tests are used for diagnosis of such conditions; they have varied sensitively and specificity, and are cumbersome to perform. We present a case of an elderly man who presented with recurrent episodes of breathlessness and chest pain, where Holter monitoring proved to be a better mode for diagnosing variant angina than more invasive provocative tests. In this report, we reemphasis the role of this simple non-invasive diagnostic mode for diagnosis of variant angina.


Electrophysiological evidence for an attentional bias in processing body stimuli in bulimia nervosa.
Empirical evidence suggests abnormalities in the processing of body stimuli in bulimia nervosa (BN). This study investigated central markers of processing body stimuli by means of event-related potentials in BN. EEG was recorded from 20 women with BN and 20 matched healthy controls while watching and evaluating underweight, normal and overweight female body pictures. Bulimics evaluated underweight bodies as less unpleasant and overweight bodies as bigger and more arousing. A higher P2 to overweight stimuli occurred in BN only. In contrast to controls, no N2 increase to underweight bodies was observed in BN. P3 was modulated by stimulus category only in healthy controls; late slow waves to underweight bodies were more pronounced in both groups. P2 amplitudes to overweight stimuli were correlated with drive for thinness and body dissatisfaction. We present novel support for altered perceptual and cognitive-affective processing of body images in BN on the subjective and electrophysiological level.


Microfluidic mixer designed for performing single-molecule kinetics with confocal detection on timescales from milliseconds to minutes.
Microfluidic mixing in combination with single-molecule spectroscopy allows the investigation of complex biomolecular processes under non-equilibrium conditions. Here we present a protocol for building, installing and operating microfluidic mixing devices optimized for this purpose. The mixer is fabricated by replica molding with polydimethylsiloxane (PDMS), which allows the production of large numbers of devices at a low cost using a single microfabricated silicon mold. The design is based on hydrodynamic focusing combined with diffusive mixing and allows single-molecule kinetics to be recorded over five orders of magnitude in time, from 1 ms to ∼100 s. Owing to microfabricated particle filters incorporated in the inlet channels, the devices provide stable flow for many hours to days without channel blockage, which allows reliable collection of high-quality data. Modular design enables rapid exchange of samples and mixing devices, which are mounted in a specifically designed holder for use with a confocal microscopy detection system. Integrated Peltier elements provide temperature control from 4 to 37 °C. The protocol includes the fabrication of a silicon master, production of the microfluidic devices, instrumentation setup and data acquisition. Once a silicon master is available, devices can be produced and experiments started within ∼1 d of preparation. We demonstrate the performance of the system with single-molecule Förster resonance energy transfer (FRET) measurements of kinetics of protein folding and conformational changes. The dead time of 1 ms, as predicted from finite element calculations, was confirmed by the measurements.


Human immunodeficiency virus type 1 viral protein R (Vpr) induces CCL5 expression in astrocytes via PI3K and MAPK signaling pathways.
Neurocognitive impairments remain prevalent in HIV-1 infected individuals despite current antiretroviral therapies. It is increasingly becoming evident that astrocytes play a critical role in HIV-1 neuropathogenesis through the production of proinflammatory cytokines/chemokines. HIV-1 viral protein R (Vpr) plays an important role in neuronal dysfunction; however, its role in neuroinflammation is not well characterized. The major objective of this study was to determine the effect of Vpr in induction of proinflammatory chemokine CCL5 in astrocytes and to define the underlying mechanism(s). SVGA astrocytes were either mock transfected or were transfected with a plasmid encoding HIV-1 Vpr, and the cells were harvested at different time intervals. The mRNA level of CCL5 expression was quantified using real-time RT-PCR, and cell culture supernatants were assayed for CCL5 protein concentration. Immunocytochemistry was performed on HIV-1 Vpr transfected astrocytes to check CCL5 expression. Various signaling mechanisms such as p38 MAPK, PI3K/Akt, NF-κB and AP-1 were explored using specific chemical inhibitors and siRNAs. HIV-1 Vpr transfected astrocytes exhibited time-dependent induction of CCL5 as compared to mock-transfected astrocytes at both the mRNA and protein level. Immunostained images of astrocytes transfected with HIV-1 Vpr also showed much higher accumulation of CCL5 in comparison to untransfected and mock-transfected astrocytes. Pre-treatment with NF-κB (SC514) and PI3K/Akt (LY294002) inhibitor partially abrogated CCL5 mRNA and protein expression levels as opposed to untreated controls after HIV-1 Vpr transfection. Specific siRNAs against p50 and p65 subunits of NF-κB, p38δ MAPK, Akt-2 and Akt-3, and AP-1 transcription factor substantially inhibited the production of CCL5 in HIV-1 Vpr transfected astrocytes. These results demonstrate the ability of HIV-1 Vpr to induce CCL5 in astrocytes in a time-dependent manner. Furthermore, this effect was observed to be mediated by transcription factors NF-κB and AP-1 and involved the p38-MAPK and PI3K/Akt pathway.


Targeted Sequencing and Meta-Analysis of Preterm Birth.
Understanding the genetic contribution(s) to the risk of preterm birth may lead to the development of interventions for treatment, prediction and prevention. Twin studies suggest heritability of preterm birth is 36-40%. Large epidemiological analyses support a primary maternal origin for recurrence of preterm birth, with little effect of paternal or fetal genetic factors. We exploited an "extreme phenotype" of preterm birth to leverage the likelihood of genetic discovery. We compared variants identified by targeted sequencing of women with 2-3 generations of preterm birth with term controls without history of preterm birth. We used a meta-genomic, bi-clustering algorithm to identify gene sets coordinately associated with preterm birth. We identified 33 genes including 217 variants from 5 modules that were significantly different between cases and controls. The most frequently identified and connected genes in the exome library were IGF1, ATM and IQGAP2. Likewise, SOS1, RAF1 and AKT3 were most frequent in the haplotype library. Additionally, SERPINB8, AZU1 and WASF3 showed significant differences in abundance of variants in the univariate comparison of cases and controls. The biological processes impacted by these gene sets included: cell motility, migration and locomotion; response to glucocorticoid stimulus; signal transduction; metabolic regulation and control of apoptosis.


A portable energy-sensitive cosmic neutron detection instrument.
The construction and testing of a portable energy-sensitive neutron instrument are described. This instrument has been designed and constructed for the primary purpose of characterizing cosmic-ray neutron fields in the upper atmosphere and in cosmic reference field facilities. The instrument comprises a helium-3 proportional counter surrounded by 15 mm of lead and 140 mm of polyethylene creating a spherical structure with a diameter of 34 cm. The instrument also incorporates 12 boron-coated diodes, six on the outside of the polyethylene layer with six placed within the structure. The dimensions, materials, and arrangement of these in the instrument have previously been optimized with the MCNPX Monte Carlo simulation software to provide a compromise between the requirements of portability and spectral response. Testing took place at several locations and experimental data from the instrument's operation at the high-altitude Jungfraujoch laboratory in the Swiss alps are presented.


Auditing SNOMED CT hierarchical relations based on lexical features of concepts in non-lattice subgraphs.
We introduce a structural-lexical approach for auditing SNOMED CT using a combination of non-lattice subgraphs of the underlying hierarchical relations and enriched lexical attributes of fully specified concept names. Our goal is to develop a scalable and effective approach that automatically identifies missing hierarchical IS-A relations. Our approach involves 3 stages. In stage 1, all non-lattice subgraphs of SNOMED CT's IS-A hierarchical relations are extracted. In stage 2, lexical attributes of fully-specified concept names in such non-lattice subgraphs are extracted. For each concept in a non-lattice subgraph, we enrich its set of attributes with attributes from its ancestor concepts within the non-lattice subgraph. In stage 3, subset inclusion relations between the lexical attribute sets of each pair of concepts in each non-lattice subgraph are compared to existing IS-A relations in SNOMED CT. For concept pairs within each non-lattice subgraph, if a subset relation is identified but an IS-A relation is not present in SNOMED CT IS-A transitive closure, then a missing IS-A relation is reported. The September 2017 release of SNOMED CT (US edition) was used in this investigation. A total of 14,380 non-lattice subgraphs were extracted, from which we suggested a total of 41,357 missing IS-A relations. For evaluation purposes, 200 non-lattice subgraphs were randomly selected from 996 smaller subgraphs (of size 4, 5, or 6) within the "Clinical Finding" and "Procedure" sub-hierarchies. Two domain experts confirmed 185 (among 223) suggested missing IS-A relations, a precision of 82.96%. Our results demonstrate that analyzing the lexical features of concepts in non-lattice subgraphs is an effective approach for auditing SNOMED CT.


Oxytocin Mediates Entrainment of Sensory Stimuli to Social Cues of Opposing Valence.
Meaningful social interactions modify behavioral responses to sensory stimuli. The neural mechanisms underlying the entrainment of neutral sensory stimuli to salient social cues to produce social learning remain unknown. We used odor-driven behavioral paradigms to ask if oxytocin, a neuropeptide implicated in various social behaviors, plays a crucial role in the formation of learned associations between odor and socially significant cues. Through genetic, optogenetic, and pharmacological manipulations, we show that oxytocin receptor signaling is crucial for entrainment of odor to social cues but is dispensable for entrainment to nonsocial cues. Furthermore, we demonstrate that oxytocin directly impacts the piriform, the olfactory sensory cortex, to mediate social learning. Lastly, we provide evidence that oxytocin plays a role in both appetitive and aversive social learning. These results suggest that oxytocin conveys saliency of social stimuli to sensory representations in the piriform cortex during odor-driven social learning.


Effect of the selective kappa-opioid receptor antagonist JDTic on nicotine antinociception, reward, and withdrawal in the mouse.
Several lines of evidence support a role for the endogenous opioid system in mediating behaviors associated with drug dependence. Specifically, recent findings suggest that the kappa-opioid receptor (KOR) may play a role in aspects of nicotine dependence, which contribute to relapse and continued tobacco smoking. The objective of this study is to determine the involvement of the KOR in the initial behavioral responses of nicotine, nicotine reward, and nicotine withdrawal using the highly selective KOR antagonist JDTic. JDTic doses of 1, 4, 8, or 16 mg/kg were administered subcutaneously (s.c.) 18 h prior to nicotine treatment. JDTic dose-dependently blocked acute nicotine-induced antinociception in the tail-flick but not the hot-plate test and did not significantly attenuate morphine's antinociceptive effect in either the tail-flick or hot-plate test. Furthermore, JDTic (8 and 16 mg/kg, s.c.) failed to block the expression of nicotine reward as measured by the conditioned place preference model. In contrast, JDTic and the KOR antagonist norBNI attenuated the expression of both the physical (somatic signs and hyperalgesia) and affective (anxiety-related behavior and conditioned place aversion) nicotine withdrawal signs. Our findings clearly show that the KOR is involved in mediating the withdrawal aspects of nicotine dependence. The results from this study suggest that blockade of the KOR by selective KOR antagonists may be useful smoking cessation pharmacotherapies.


Personality characteristics in a Swedish national sample of identifiable oocyte donors.
To study the personality characteristics of identifiable oocyte donors in a national sample in comparison with normal values. Descriptive study. All Swedish donation programmes. In total, 181 women out of 221 donors recruited during 2005-2008. Standardised questionnaires were used to measure personality characteristics. Demographics, temperament and character inventory (TCI). The majority (69%) of the donors had biological children of their own. The results from the TCI indicate that the oocyte donors were all within the normal range of character. With regard to personality, a significant difference was evident between the two groups: oocyte donors showed lower means for harm avoidance and higher scores for persistence than the controls. This indicates that the donors felt less worried, and displayed a lower level of fear of uncertainty, shyness and fatiguability, and a higher level of persistence, than the controls. In the present sample, 29 (16%) of the donors were so-called 'known donors', that is the recipient couples and the donors were known to each other. 'Known donors' displayed a mature and stable character. We found that the women who had been accepted for inclusion in this nationwide oocyte donor programme were all well adjusted and mature.


Rhesus macaque θ-defensin RTD-1 inhibits proinflammatory cytokine secretion and gene expression by inhibiting the activation of NF-κB and MAPK pathways.
θ-Defensins are pleiotropic, macrocyclic peptides that are expressed uniquely in Old World monkeys. The peptides are potent, broad-spectrum microbicides that also modulate inflammatory responses in vitro and in animal models of viral infection and polymicrobial sepsis. θ-Defensins suppress proinflammatory cytokine secretion by leukocytes stimulated with diverse Toll-like receptor (TLR) ligands. Studies were performed to delineate anti-inflammatory mechanisms of rhesus θ-defensin 1 (RTD-1), the most abundant θ-defensin isoform in macaque granulocytes. RTD-1 reduced the secretion of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-8 in lipopolysaccharide (LPS)-stimulated human blood monocytes and THP-1 macrophages, and this was accompanied by inhibition of nuclear factor κB (NF-κB) activation and mitogen-activated protein kinase (MAPK) pathways. Peptide inhibition of NF-κB activation occurred following stimulation of extracellular (TLRs 1/2 and 4) and intracellular (TLR9) receptors. Although RTD-1 did not inhibit MAPK in unstimulated cells, it induced phosphorylation of Akt in otherwise untreated monocytes and THP-1 cells. In the latter, this occurred within 10 min of RTD-1 treatment and produced a sustained elevation of phosphorylated Akt (pAkt) for at least 4 h. pAkt is a negative regulator of MAPK and NF-κB activation. RTD-1 inhibited IκBα degradation and p38 MAPK phosphorylation, and stimulated Akt phosphorylation in LPS-treated human primary monocytes and THP-1 macrophages. Specific inhibition of phosphatidylinositol 3-kinase (PI3K) blocked RTD-1-stimulated Akt phosphorylation and reversed the suppression of NF-κB activation by the peptide. These studies indicate that the anti-inflammatory properties of θ-defensins are mediated by activation of the PI3K/Akt pathway and suppression of proinflammatory signals in immune-stimulated cells.


Notch ligands regulate the muscle stem-like state ex vivo but are not sufficient for retaining regenerative capacity.
Myogenic stem cells are a promising avenue for the treatment of muscular disorders. Freshly isolated muscle stem cells have a remarkable engraftment ability in vivo, but their cell number is limited. Current conventional culture conditions do not allow muscle stem cells to expand in vitro with their bona fide engraftment efficiency, requiring the improvement of culture procedures for achieving successful cell-therapy for muscle disorders. Here we expanded mouse muscle stem cells and human myoblasts with Notch ligands, DLL1, DLL4, and JAG1 to activate Notch signaling in vitro and to investigate whether these cells could retain their engraftment efficiency. Notch signaling promotes the expansion of Pax7+MyoD- mouse muscle stem-like cells and inhibits differentiation even after passage in vitro. Treatment with Notch ligands induced the Notch target genes and generated PAX7+MYOD- stem-like cells from human myoblasts previously cultured on conventional culture plates. However, cells treated with Notch ligands exhibit a stem cell-like state in culture, yet their regenerative ability was less than that of freshly isolated cells in vivo and was comparable to that of the control. These unexpected findings suggest that artificial maintenance of Notch signaling alone is insufficient for improving regenerative capacity of mouse and human donor-muscle cells and suggest that combinatorial events are critical to achieve muscle stem cell and myoblast engraftment potential.


Biological markers for anxiety disorders, OCD and PTSD: A consensus statement. Part II: Neurochemistry, neurophysiology and neurocognition.
Biomarkers are defined as anatomical, biochemical or physiological traits that are specific to certain disorders or syndromes. The objective of this paper is to summarise the current knowledge of biomarkers for anxiety disorders, obsessive-compulsive disorder (OCD) and posttraumatic stress disorder (PTSD). Findings in biomarker research were reviewed by a task force of international experts in the field, consisting of members of the World Federation of Societies for Biological Psychiatry Task Force on Biological Markers and of the European College of Neuropsychopharmacology Anxiety Disorders Research Network. The present article (Part II) summarises findings on potential biomarkers in neurochemistry (neurotransmitters such as serotonin, norepinephrine, dopamine or GABA, neuropeptides such as cholecystokinin, neurokinins, atrial natriuretic peptide, or oxytocin, the HPA axis, neurotrophic factors such as NGF and BDNF, immunology and CO2 hypersensitivity), neurophysiology (EEG, heart rate variability) and neurocognition. The accompanying paper (Part I) focuses on neuroimaging and genetics. Although at present, none of the putative biomarkers is sufficient and specific as a diagnostic tool, an abundance of high quality research has accumulated that should improve our understanding of the neurobiological causes of anxiety disorders, OCD and PTSD.


A transcript profiling approach reveals the zinc finger transcription factor ZNF191 is a pleiotropic factor.
The human zinc finger protein 191 (ZNF191) is a member of the SCAN domain family of Krüppel-like zinc finger transcription factors. ZNF191 shows 94% identity to its mouse homologue zinc finger protein 191(Zfp191), which is the most highly conserved among the human-mouse SCAN family member orthologues pairs. Zfp191 is widely expressed during early embryogenesis and in adult organs. Moreover, Zfp191-/- embryos have been shown to be severely retarded in development and die approximately at embryonic day E7.5. ZNF191 can specifically interact with the widespread TCAT motif which constitutes the HUMTH01 microsatellite in the tyrosine hydroxylase (TH) gene. Allelic variations of HUMTH01 have been stated to have a quantitative silencing effect on TH gene expression and to correlate with quantitative and qualitative changes in the binding by ZNF191. In addition, ZNF191 displays a suppressive effect on the transcription; however, little downstream targets have identified. We searched for ZNF191 target genes by using a transient overexpression and knockdown strategy in the human embryo kidney (HEK293) cells. Microarray analyses identified 6094 genes modulated by overexpression of ZNF191 and 3332 genes regulated by knockdown of ZNF191, using a threshold of 1.2-fold. Several interested candidate genes, validated by real time RT-PCR, were correlated well with the array data. Interestingly, 1456 genes were identified in both transient overexpression and transient knockdown strategies. The GenMAPP and MappFinder software packages were further used for pathway analysis of these significantly altered genes. Several gene pathways were found to be involved in processes of the regulation of kinase activity, transcription, angiogenesis, brain development and response to DNA damage. Our analysis reveals for the first time that ZNF191 is a pleiotropic factor that has a role in hematopoiesis, brain development and cancers.


Phase I dose-escalation study of AZD7762, a checkpoint kinase inhibitor, in combination with gemcitabine in US patients with advanced solid tumors.
AZD7762 is a Chk1 kinase inhibitor which increases sensitivity to DNA-damaging agents, including gemcitabine. We evaluated the safety of AZD7762 monotherapy and with gemcitabine in advanced solid tumor patients. In this Phase I study, patients received intravenous AZD7762 on days 1 and 8 of a 14-day run-in cycle (cycle 0; AZD7762 monotherapy), followed by AZD7762 plus gemcitabine 750-1,000 mg/m(2) on days 1 and 8, every 21 days, in ascending AZD7762 doses (cycle 1; combination therapy). Forty-two patients received AZD7762 6 mg (n = 9), 9 mg (n = 3), 14 mg (n = 6), 21 mg (n = 3), 30 mg (n = 7), 32 mg (n = 6), and 40 mg (n = 8), in combination with gemcitabine. Common adverse events (AEs) were fatigue [41 % (17/42) patients], neutropenia/leukopenia [36 % (15/42) patients], anemia/Hb decrease [29 % (12/42) patients] and nausea, pyrexia and alanine aminotransferase/aspartate aminotransferase increase [26 % (11/42) patients each]. Grade ≥3 AEs occurred in 19 and 52 % of patients in cycles 0 and 1, respectively. Cardiac dose-limiting toxicities occurred in two patients (both AZD7762 monotherapy): grade 3 troponin I increase (32 mg) and grade 3 myocardial ischemia with chest pain, electrocardiogram changes, decreased left ventricular ejection fraction, and increased troponin I (40 mg). AZD7762 exposure increased linearly. Gemcitabine did not affect AZD7762 pharmacokinetics. Two non-small-cell lung cancer patients achieved partial tumor responses (AZD7762 6 mg/gemcitabine 750 mg/m(2) and AZD7762 9 mg cohort). The maximum-tolerated dose of AZD7762 in combination with gemcitabine 1,000 mg/m(2) was 30 mg. Although development of AZD7762 is not going forward owing to unpredictable cardiac toxicity, Chk1 remains an important therapeutic target.


Acinetobacter baumannii IC2 and IC5 Isolates with Co-Existing blaOXA-143-like and blaOXA-72 and Exhibiting Strong Biofilm Formation in a Mexican Hospital.
Acinetobacter baumannii is an opportunistic pathogen responsible for healthcare-associated infections (HAIs) and outbreaks. Antimicrobial resistance mechanisms and virulence factors allow it to survive and spread in the hospital environment. However, the molecular mechanisms of these traits and their association with international clones are frequently unknown in low- and middle-income countries. Here, we analyze the phenotype and genotype of seventy-six HAIs and outbreak-causing A. baumannii isolates from a Mexican hospital over ten years, with special attention to the carbapenem resistome and biofilm formation. The isolates belonged to the global international clone (IC) 2 and the Latin America endemic IC5 and were predominantly extensively drug-resistant (XDR). Oxacillinases were identified as a common source of carbapenem resistance. We noted the presence of the blaOXA-143-like family (not previously described in Mexico), the blaOXA-72 and the blaOXA-398 found in both ICs. A low prevalence of efflux pump overexpression activity associated with carbapenem resistance was observed. Finally, strong biofilm formation was found, and significant biofilm-related genes were identified, including bfmRS, csuA/BABCDE, pgaABCD and ompA. This study provides a comprehensive profile of the carbapenem resistome of A. baumannii isolates belonging to the same pulse type, along with their significant biofilm formation capacity. Furthermore, it contributes to a better understanding of their role in the recurrence of infection and the endemicity of these isolates in a Mexican hospital.


Nanoprobe-based force spectroscopy as a versatile platform for probing the mechanical adhesion of bacteria.
The first stage of biofilm-associated infections is commonly caused by initial adhesion of bacteria to intravascular tubes, catheters and other medical devices. The overuse of antibiotics to treat these infections has led to the spread of antibiotic resistance, which has made infections difficult to eradicate. It is crucial to develop advanced strategies to inhibit biofilm formation, avoiding the emergence of antibiotic resistance. Previously, it has been reported that substrate stiffness plays an important role in the initial attachment of bacteria. However, the mechanism of how the stiffness modulates the initial adhesion of bacteria remains unclear. Here, we developed magnetic nanoprobe-based force-induced remnant magnetization spectroscopy (FIRMS) as a new platform to measure the adhesion force of bacteria. Through examining the initial adhesion force and the adhesive protein, fibronectin-binding protein (FnBP), of Staphylococcus aureus (S. aureus), we found that the increase of the substrate stiffness promoted the expression of FnBP, thus enhancing the initial adhesion force of bacteria. Following the formation of initial adhesion, the substrates with soft stiffness delayed the biofilm formation, whereas those with moderate stiffness showed preferential promotion of the biofilm formation. We expect this versatile platform to be beneficial to the study of adhesion behaviors of bacteria that sheds light on the design of new medical materials to treat microbial infections.


All-optical manipulation of singlet exciton transport in individual supramolecular nanostructures by triplet gating.
Directed transport of singlet excitation energy is a key process in natural light-harvesting systems and a desired feature in assemblies of functional organic molecules for organic electronics and nanotechnology applications. However, progress in this direction is hampered by the lack of concepts and model systems. Here we demonstrate an all-optical approach to manipulate singlet exciton transport pathways within supramolecular nanostructures via singlet-triplet annihilation, i.e., to enforce an effective motion of singlet excitons along a predefined direction. For this proof-of-concept, we locally photo-generate a long-lived triplet exciton population and subsequently a singlet exciton population on single bundles of H-type supramolecular nanofibres using two temporally and spatially separated laser pulses. The local triplet exciton population operates as a gate for the singlet exciton transport since singlet-triplet annihilation hinders singlet exciton motion across the triplet population. We visualize this manipulation of singlet exciton transport via the fluorescence signal from the singlet excitons, using a detection-beam scanning approach combined with time-correlated single-photon counting. Our reversible, all-optical manipulation of singlet exciton transport can pave the way to realising new design principles for functional photonic nanodevices.


Methodology of the Stroke Self-Management Rehabilitation Trial: an international, multisite pilot trial.
Stroke is a major cause of long-term adult disability with many survivors living in the community relying on family members for on-going support. However, reports of inadequate understanding of rehabilitation techniques are common. A self-management DVD-based observational learning tool may help improve functional outcomes for survivors of stroke and reduce caregivers' burden. This article describes the methodology of the stroke self-management rehabilitation trial. The overall aim of this pilot trial is to assess the feasibility and preliminary efficacy of a DVD-based intervention for improving functional outcomes of survivors of stroke 2 months postrandomization to inform the design of a full-scale randomized clinical trial. Recruitment of a minimum of 20 survivors of stroke and their informal caregivers (where available) in each of the participating centers will occur across multiple international sites. After baseline assessments, participants will be randomly assigned to an intervention or standard care group. The intervention comprises a structured DVD observation and practice schedule over 8 weeks. All participants will complete follow-up assessments. The outcome measures will include a global shift in the Rankin Scale scores and dichotomized scores, changes in quality of life, general health, depression, and caregiver burden at 2 months postrandomization. A qualitative analysis of the effects of the intervention will also be undertaken. The results of the pilot study will provide knowledge of whether observational learning techniques delivered via DVD can effectively improve recovery after stroke and reduce caregiver burden.


The challenge of rapid management of fever in children under 5 in Guinea.
In 2013, the Guinean health authority had to reorganise and run a national response against malaria as a priority. The review of the National Strategic Plan to fight malaria in Guinea was carried out and one of its critical components was the prevention and rapid management of fever (RMF) attributable to malaria in children. The study reports on the demographic and health determinants of this rapid management in children under 5. The participants were 4786 children from 2874 representative households. RMF was defined in terms of recourse to primary care. The recourse was defined by child's reference for the treatment of fever which led or not to treatment of malaria. We found that 1491 children (31.2%) had a bout of fever within the 2 weeks that preceded the survey. The prevalence of malaria was 45.4% among those children who have a bout of fever. The recourse to traditional healers was estimated at 9.6% and the use of health facilities was estimated at 71.5%. Overall, 74.9% of children with fever received treatment within the recommended timeliness (24 h), with regional disparity in this rapid response. The high proportion of recourse to traditional healers is still a matter of concern. New control and prevention strategies should be extended to traditional healers for their training and involvement in directing febrile children to health facilities.


Mathematical modeling and parametrical analysis of the temperature dependency of control drug release from biodegradable nanoparticles.
In this study we describe a mathematical analysis that considers the temperature effects of the controlled drug release process from biodegradable poly-d,l-lactide-co-glycolide (PLGA) nanoparticles. Temperature effects are incorporated and applied to two drug release models. The first one consists of a two-stage release process that considers only simultaneous contributions of initial burst and nanoparticle degradation-relaxation (BR model). The second one is a three release stage model that considers, additionally, a simultaneous drug diffusion (BRD model) step. In these models, the temperature dependency of the release parameters, initial burst constant, k b, the rate of degradation-relaxation constant, k r, time to achieve 50% of release, t max, and effective diffusion coefficient constant (D e), are determined using mathematical expressions analogous to the Arrhenius equation. The temperature dependent models are used to analyze the release of previously encapsulated Rhodamine 6G dye as a model drug in polyethylene glycol modified PLGA nanoparticles. The experimental data used to develop the mathematical model was obtained from release studies carried out in phosphate buffer pH 7.4 at 37 °C, 47 °C, and 57 °C. Multiphasic release behaviors with an overall increase rate associated with the incubation temperature were observed. The study incorporates a parametrical analysis that can evaluate diverse temperature variation effects of the controlled release parameters for the two models.


Awards for Distinguished Scientific Contributions.
The Awards for Distinguished Scientific Contributions are presented to persons who, in the opinion of the Committee on Scientific Awards, have made distinguished theoretical or empirical contributions to basic research in psychology. The 2018 recipients of the APA Scientific Contribution Awards were recognized by the 2017 Board of Scientific Affairs and selected by the 2017 Committee on Scientific Awards. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Laminin chains in developing and adult human myotendinous junctions.
In addition to being the specialized site for transmission of force from the muscle to the tendon, the myotendinous junction (MTJ) also plays an important role in muscle splitting during morphogenesis. An early event in the formation of the MTJ is a regional deposition of basement membranes. We used immunocytochemistry to investigate the distribution of laminin chains during the development of MTJs in human limb muscle at 8-22 weeks of gestation (wg) and in adult MTJs. We used polyclonal antibodies and a new monoclonal antibody (MAb) against the human laminin alpha1 G4/G5 domains. At 8-10 wg, laminin alpha1 and laminin alpha5 chains were specifically localized to the MTJ. Laminin alpha1 chain remained restricted to the MTJ at 22 wg as the laminin beta2 chain had appeared, whereas the laminin alpha5 chain became deposited along the entire length of the myotubes from 12 wg. In the adult MTJ, only vestigial amounts of laminin alpha1 and laminin alpha5 chains could be detected. On the basis of co-distribution data, we speculate that laminin alpha1 chain in the forming MTJ undergoes an isoform switch from laminin 1 to laminin 3. Our data indicate a potentially important role for laminin alpha1 chain in skeletal muscle formation. (J Histochem Cytochem 48:201-209, 2000)


Larval green and white sturgeon swimming performance in relation to water-diversion flows.
Little is known of the swimming capacities of larval sturgeons, despite global population declines in many species due in part to fragmentation of their spawning and rearing habitats by man-made water-diversion structures. Larval green (Acipenser medirostris) and white sturgeon (Acipenser transmontanus) inhabit the highly altered Sacramento-San Joaquin watershed, making them logical species to examine vulnerability to entrainment by altered water flows. The risk of larval sturgeon entrainment is influenced by the ontogeny of swimming capacity and dispersal timing and their interactions with water-diversion structure operations. Therefore, the aim of this study was to describe and compare the ontogeny and allometry of larval green and white sturgeon swimming capacities until completion of metamorphosis into juveniles. Despite the faster growth rates and eventual larger size of larval white sturgeon, green sturgeon critical swimming velocities remained consistently, though modestly, greater than those of white sturgeon throughout the larval life stage. Although behavioural interactions with water-diversion structures are also important considerations, regarding swimming capacity, Sacramento-San Joaquin sturgeons are most vulnerable to entrainment in February-May, when white sturgeon early larvae are in the middle Sacramento River, and April-May, when green sturgeon early larvae are in the upper river. Green sturgeon migrating downstream to the estuary and bays in October-November are also susceptible to entrainment due to their movements combined with seasonal declines in their swimming capacity. An additional inter-species comparison of the allometric relationship between critical swimming velocities and total length with several sturgeon species found throughout the world suggests a similar ontogeny of swimming capacity with growth. Therefore, although dispersal and behaviour differ among river systems and sturgeon species, similar recommendations are applicable for managers seeking to balance water demands with restoration and conservation of sturgeons worldwide.


Pharmacokinetic properties and interactions with blood components of N4-hexadecyl-1-beta-D-arabinofuranosylcytosine (NHAC) incorporated into liposomes.
N4-Hexadecyl-1-beta-D-arabinofuranosylcytosine (NHAC) is a new lipophilic derivative of 1-beta-D-arabinofuranosylcytosine (ara-C) with strong antitumour activity. The interactions of NHAC incorporated into small unilamellar liposomes of different compositions with blood components were evaluated. In comparison with ara-C, NHAC is highly protected against deamination to inactive arabinofuranosyluracil (ara-U) in human plasma, resulting in only 2% conversion into ara-U after 4 h incubation at 37 degrees C, whereas from ara-C more than 80% was deaminated. In in-vitro incubations with human blood, it was found that NHAC was transferred from the liposomes at about 47% efficiency to plasma proteins, particularly to albumin and to the high and low density lipoproteins. The remaining part of NHAC was bound to erythrocytes (50%) and to leucocytes (3%). The addition of poly(ethylene) glycol-modified phospholipids to the liposomes (PEG liposomes), which were composed of soy phosphatidylcholine and cholesterol (plain liposomes), did not significantly prevent the fast transfer of NHAC from the liposomes to the blood components. Pharmacokinetic studies in mice revealed that NHAC had biphasic kinetics in blood with a t1/2 alpha of 16 min and a t1/2 beta of 3.8 h when the drug was formulated in plain liposomes and a t1/2 alpha of 15 min and a t1/2 beta of 9.67 h in PEG liposomes, respectively. NHAC was predominantly distributed in the liver with 29% of the injected dose found after 30 min.(ABSTRACT TRUNCATED AT 250 WORDS)


Improving the neurodevelopmental outcomes of low-birthweight infants.
Infants born with low birthweight (LBW) have poorer neurodevelopmental outcomes compared with their term counterparts with appropriate weight for gestational age. The perinatal period is a time of high energy and high nutrient needs, and any process, such as preterm birth, poor nutrition or placental insufficiency, that interrupts the concentrated flow of nutrients to the fetus may result in babies with LBW. Therefore, it makes logical sense that at least part of the cognitive deficits may be explained by nutritional deprivation. The nutrients commonly deficient in LBW infants include protein and energy and micronutrients such as iron, zinc and long chain polyunsaturated fatty acids. In this review, we aimed to determine the effect of nutrient supplementation on neurodevelopment in LBW infants. While few trials have supported the hypothesis that nutritional supplementation improves neurodevelopment, many studies are limited by sample size and methodological shortcomings. Further large-scale rigorously designed intervention trials, with long-term neurodevelopment follow-up, are required to determine the optimal nutritional supplements and the timing of their administration to LBW infants.


Detection of arecoline by simple high-performance thin-layer chromatographic method in Indian nontobacco pan masala.
Chewing the habit of blended pan masala containing areca nut with or without tobacco is a common practice in the Indian subcontinent. Arecoline, a pyridine alkaloid presence in areca nut alarmed for oral carcinogenesis and strictly prohibited in the western world. However, in India using blended pan masala is very popular among young and old individuals. In this context, we aimed to detect arecoline in Indian blended nontobacco pan masala sold in Kolkata using a simple densitometric high-performance thin-layer chromatographic (HPTLC) method and for alarming their use in common people. Eleven popularly Indian blended nontobacco pan masala were collected from the territory of Kolkata and isolated arecoline, following solvent extraction method derived for pyridine alkaloid. The quantitative analysis of arecoline was measured using automated software-based HPTLC instruments and validated the method according to International Conference on Harmonization guidelines. Arecoline was detected in all 11 blended nontobacco pan masala samples in a range of minimum 130 to maximum 415 μg/g dry samples. Arecoline is hazardous carcinogenic compound, so the use of Indian blended nontobacco pan masala should be restricted. Further, the method was found suitable for routine quantitative analysis of arecoline in areca nut containing substances.


Portrait of the potential barrier at metal-organic nanocontacts.
Electron transport through metal-molecule contacts greatly affects the operation and performance of electronic devices based on organic semiconductors and is at the heart of molecular electronics exploiting single-molecule junctions. Much of our understanding of the charge injection and extraction processes in these systems relies on our knowledge of the potential barrier at the contact. Despite significant experimental and theoretical advances a clear rationale of the contact barrier at the single-molecule level is still missing. Here, we use scanning tunnelling microscopy to probe directly the nanocontact between a single molecule and a metal electrode in unprecedented detail. Our experiments show a significant variation on the submolecular scale. The local barrier modulation across an isolated 4-[trans-2-(pyrid-4-yl-vinyl)] benzoic acid molecule bound to a copper(111) electrode exceeds 1 eV. The giant modulation reflects the interaction between specific molecular groups and the metal and illustrates the critical processes determining the interface potential. Guided by our results, we introduce a new scheme to locally manipulate the potential barrier of the molecular nanocontacts with atomic precision.


The Advanced Glaucoma Intervention Study (AGIS): 13. Comparison of treatment outcomes within race: 10-year results.
To present for black and white patients with medically uncontrolled glaucoma 10-year results of treatment with 1 of 2 randomly assigned surgical intervention sequences. Randomized clinical trial. Three hundred thirty-two black patients (451 eyes) and 249 white patients (325 eyes). Eyes had glaucoma that could not be controlled with medications alone. Eyes were randomly assigned to 1 of 2 sequences: argon laser trabeculoplasty (ALT)-trabeculectomy-trabeculectomy (ATT) or trabeculectomy-ALT-trabeculectomy (TAT). Second and third interventions were offered after failure of the preceding intervention. Minimum required intraocular pressure (IOP) for intervention failure ranged upward from 18 mmHg, the value depending on whether recent optic disc or visual field (VF) deterioration occurred, and on the magnitude of the field defect. Patients were observed every 6 months, with total potential follow-up ranging from 8 years, 4 months to 13 years. The averages over follow-up of (1) the percentage of eyes having moderate loss of VF and (2) the percentage of eyes having moderate loss of visual acuity (VA). Race-treatment interactions in VF and VA loss are significant for the 2 main outcome measures; therefore, results of treatment sequence differences are presented by race. In black patients the average percent of eyes with VF loss was less in the ATT sequence than in the TAT sequence, a difference that is not statistically significant at any visit. In white patients, conversely, after 18 months the average percent of eyes with VF loss was less in the TAT sequence, a difference that increases and is statistically significant in years 8 to 10. In both black and white patients, the average percent of eyes with VA loss was less in the ATT sequence; this difference is statistically significant throughout 10 follow-up years in black patients and is statistically significant only for the first year in white patients. In both black and white patients, average IOP reductions were greater in the TAT sequence, though the TAT-ATT difference was substantially greater in white patients. In both black and white patients, first-intervention failure rates were substantially lower for trabeculectomy than for trabeculoplasty. Ten-year cumulative incidence of unilateral VF impairment comparable to legal blindness was modest in eyes of black (ATT 11.9%, TAT 18.5%) and white (ATT 9.9%, TAT 7.3%) patients. Although IOP was lowered in both sequences in black and white patients with medically uncontrolled glaucoma, long-term visual function outcomes were better for the ATT sequence in black patients and better for the TAT sequence in white patients.


Effects of Oxidative Stress and Testosterone on Pro-Inflammatory Signaling in a Female Rat Dopaminergic Neuronal Cell Line.
Parkinson's disease, a progressive neurodegenerative disorder, is associated with oxidative stress and neuroinflammation. These pathological markers can contribute to the loss of dopamine neurons in the midbrain. Interestingly, men have a 2-fold increased incidence for Parkinson's disease than women. Although the mechanisms underlying this sex difference remain elusive, we propose that the primary male sex hormone, testosterone, is involved. Our previous studies show that testosterone, through a putative membrane androgen receptor, can increase oxidative stress-induced neurotoxicity in dopamine neurons. Based on these results, this study examines the role of nuclear factor κ B (NF-κB), cyclooxygenase-2 (COX2), and apoptosis in the deleterious effects of androgens in an oxidative stress environment. We hypothesize, under oxidative stress environment, testosterone via a putative membrane androgen receptor will exacerbate oxidative stress-induced NF-κB/COX2 signaling in N27 dopaminergic neurons, leading to apoptosis. Our data show that testosterone increased the expression of COX2 and apoptosis in dopamine neurons. Inhibiting the NF-κB and COX2 pathway with CAPE and ibuprofen, respectively, blocked testosterone's negative effects on cell viability, indicating that NF-κB/COX2 cascade plays a role in the negative interaction between testosterone and oxidative stress on neuroinflammation. These data further support the role of testosterone mediating the loss of dopamine neurons under oxidative stress conditions, which may be a key mechanism contributing to the increased incidence of Parkinson's disease in men compared with women.


Database Extraction of Metabolite Information of Drug Candidates: Analysis of 27 AstraZeneca Compounds with Human Absorption, Distribution, Metabolism, and Excretion Data.
As part of the drug discovery and development process, it is important to understand the human metabolism of a candidate drug prior to clinical studies. Preclinical in vitro and in vivo experiments across species are conducted to build knowledge concerning human circulating metabolites in preparation for clinical studies; therefore, the quality of these experiments is critical. Within AstraZeneca, all metabolite identification (Met-ID) information is stored in a global database using ACDLabs software. In this study, the Met-ID information derived from in vitro and in vivo studies for 27 AstraZeneca drug candidates that underwent human absorption, distribution, metabolism, and excretion studies was extracted from the database. The retrospective analysis showed that 81% of human circulating metabolites were previously observed in preclinical in vitro and/or in vivo experiments. A detailed analysis was carried out to understand which human circulating metabolites were not captured in the preclinical experiments. Metabolites observed in human hepatocytes and rat plasma but not seen in circulation in humans (extraneous metabolites) were also investigated. The majority of human specific circulating metabolites derive from multistep biotransformation reactions that may not be observed in in vitro studies within the limited time frame in which cryopreserved hepatocytes are active. Factors leading to the formation of extraneous metabolites in preclinical studies seemed to be related to species differences with respect to transporter activity, secondary metabolism, and enzyme kinetics. This retrospective analysis assesses the predictive value of Met-ID experiments and improves our ability to discriminate between metabolites expected to circulate in humans and irrelevant metabolites seen in preclinical studies.


Genotoxicity and Cytotoxicity of Cadmium Sulfide Nanomaterials to Mice: Comparison Between Nanorods and Nanodots.
Cadmium sulfide (CdS) nanomaterials (such as CdS nanodots or nanorods) are widely used in optical, electronic, and biological applications. Large-scale production and use of these materials will likely result in accidental and incidental releases, which raise concerns about their potential environmental and human-health impacts. Most studies on toxicity of Cd-containing nanomaterials have focused on nanodots, and the relative toxicity of Cd-containing nanorods is not well understood. Here, we compared genotoxicity and cytotoxicity of CdS nanorods (30-50 nm diameter, 500-1100 nm length) and cubic CdS nanodots (3-5 nm) in mice by examining total cadmium accumulation in organs, acute toxicity, DNA damage, spermatozoon viability and abnormality, kidney and liver damage, and oxidative stress. Compared with (smaller) nanodots, nanorods resulted in relatively low bioaccumulation, acute toxicity, and damage to spermatozoa and the tested organs. Differences in toxicity between CdS nanodots and nanorods could not be fully explained by differences in their metal ion (Cd2+) release patterns, based on control tests with mice gavaged with dissolved CdCl2 at equivalent concentrations. This underscores that toxicity of metallic nanomaterials could not be solely predicted based either on their elemental composition or on the amount of ions released before receptor intake. Particle morphology (including size) may also need to be considered.


Formation of biodegradable microcapsules utilizing 3D, selectively surface-modified PDMS microfluidic devices.
We have successfully demonstrated the formation of biodegradable microcapsules utilizing PDMS double-emulsification devices. Specially designed 3D PDMS microchannels with surfaces selectively modified by a self-aligned photografting process are employed to generate monodisperse water-in-organic-solvent-in-water (W/O/W) emulsions in a controlled manner. Mainly by varying the outer and inner fluid flow-rates, the dimensions of resulting double emulsions can be adjusted as desired. Meanwhile, biodegradable materials are dissolved in the middle organic solvent (in this work ethyl acetate is used), and solidified into microcapsules once the solvent is extracted. In the prototype demonstration, microcapsules made up of poly(L-lactic acid), trilaurin, and phosphocholine were successfully fabricated. In addition, it was also demonstrated that gamma-Fe(2)O(3) nanoparticles can be simultaneously embedded into the microcapsules, which consequently become responsive to electromagnetic stimulation. As such, the presented PDMS microfluidic devices could potentially serve as versatile encapsulation apparatus, and the fabricated biodegradable microcapsules could function as controlled delivery systems, which are desired for a variety of biological and pharmaceutical applications.