Cell signalling of glucagon-like peptide-1 action in rat skeletal muscle.
Glucagon-like peptide-1 (GLP-1), an incretin with glucose-dependent insulinotropic and insulin-independent antidiabetic properties, has insulin-like effects on glucose metabolism in extrapancreatic tissues participating in overall glucose homeostasis. These effects are exerted through specific receptors not associated with cAMP, an inositol phosphoglycan being a possible second messenger. In rat hepatocytes, activation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB), protein kinase C (PKC) and protein phosphatase 1 (PP-1) has been shown to be involved in the GLP-1-induced stimulation of glycogen synthase. We have investigated the role of enzymes known or suggested to mediate the actions of insulin in the GLP-1-induced increase in glycogen synthase a activity in rat skeletal muscle strips. We first explored the effect of GLP-1, compared with that of insulin, on the activation of PI3K, PKB, p70s6 kinase (p70s6k) and p44/42 mitogen-activated protein kinases (MAPKs) and the action of specific inhibitors of these kinases on the insulin- and GLP-1-induced increment in glycogen synthase a activity. The study showed that GLP-1, like insulin, activated PI3K/PKB, p70s6k and p44/42. Wortmannin (a PI3K inhibitor) reduced the stimulatory action of insulin on glycogen synthase a activity and blocked that of GLP-1, rapamycin (a 70s6k inhibitor) did not affect the action of GLP-1 but abolished that of insulin, PD98059 (MAPK inhibitor) was ineffective on insulin but blocked the action of GLP-1, okadaic acid (a PP-2A inhibitor) and tumour necrosis factor-alpha (a PP-1 inhibitor) were both ineffective on GLP-1 but abolished the action of insulin, and Ro 31-8220 (an inhibitor of some PKC isoforms) reduced the effect of GLP-1 while completely preventing that of insulin. It was concluded that activation of PI3K/PKB and MAPKs is required for the GLP-1-induced increment in glycogen synthase a activity, while PKC, although apparently participating, does not seem to play an essential role; unlike in insulin signaling, p70s6k, PP-1 and PP-2A do not seem to be needed in the action of GLP-1 upon glycogen synthase a activity in rat muscle.


Transforming growth factor-beta-induced growth inhibition in a Smad4 mutant colon adenoma cell line.
Transforming growth factor-beta (TGF-beta) inhibits growth and induces apoptosis of colon epithelial cells. Binding of TGF-beta to its receptor induces phosphorylation of the Smad proteins Smad2 and Smad3, which then form heteromeric complexes with Smad4, translocate to the nucleus, and activate gene transcription. Smad4 function has been considered an obligate requirement for TGF-beta signaling, and Smad4 mutations present in some cancers have been considered sufficient to inactivate TGF-beta signaling. In this work, we describe studies with a nontransformed human colon epithelial cell line that is mutant for Smad4 but remains growth-inhibited by TGF-beta. The colon cell line VACO-235 has lost one of its Smad4 alleles via a chromosome 18q deletion. The remaining allele bears two missense point mutations located in regions important for Smad4 trimer formation, which is thought necessary for Smad4 function. As expected, pSBE4-BV/Luc, a Smad4-activated transcriptional reporter, was inactive in VACO-235. Nonetheless, VACO-235 demonstrated 80% growth inhibition in response to TGF-beta, as well as retention of some TGF-beta-mediated activation of the p3TP-Lux transcriptional reporter. Transient transfection of the VACO-235 Smad4 mutant allele into a Smad4-null cell line confirmed that this allele is functionally inactive as assayed by both the pSBE4-BV and p3TP-Lux reporters. The simplest explanation of these results is that there is a non-Smad4-dependent pathway for TGF-beta-mediated signaling and growth inhibition in VACO-235 cells.


An NK Cell Perforin Response Elicited via IL-18 Controls Mucosal Inflammation Kinetics during Salmonella Gut Infection.
Salmonella Typhimurium (S.Tm) is a common cause of self-limiting diarrhea. The mucosal inflammation is thought to arise from a standoff between the pathogen's virulence factors and the host's mucosal innate immune defenses, particularly the mucosal NAIP/NLRC4 inflammasome. However, it had remained unclear how this switches the gut from homeostasis to inflammation. This was studied using the streptomycin mouse model. S.Tm infections in knockout mice, cytokine inhibition and -injection experiments revealed that caspase-1 (not -11) dependent IL-18 is pivotal for inducing acute inflammation. IL-18 boosted NK cell chemoattractants and enhanced the NK cells' migratory capacity, thus promoting mucosal accumulation of mature, activated NK cells. NK cell depletion and Prf-/- ablation (but not granulocyte-depletion or T-cell deficiency) delayed tissue inflammation. Our data suggest an NK cell perforin response as one limiting factor in mounting gut mucosal inflammation. Thus, IL-18-elicited NK cell perforin responses seem to be critical for coordinating mucosal inflammation during early infection, when S.Tm strongly relies on virulence factors detectable by the inflammasome. This may have broad relevance for mucosal defense against microbial pathogens.


Genomic modulators of gene expression in human neutrophils.
Neutrophils form the most abundant leukocyte subset and are central to many disease processes. Technical challenges in transcriptomic profiling have prohibited genomic approaches to date. Here we map expression quantitative trait loci (eQTL) in peripheral blood CD16+ neutrophils from 101 healthy European adults. We identify cis-eQTL for 3281 neutrophil-expressed genes including many implicated in neutrophil function, with 450 of these not previously observed in myeloid or lymphoid cells. Paired comparison with monocyte eQTL demonstrates nuanced conditioning of genetic regulation of gene expression by cellular context, which relates to cell-type-specific DNA methylation and histone modifications. Neutrophil eQTL are markedly enriched for trait-associated variants particularly autoimmune, allergy and infectious disease. We further demonstrate how eQTL in PADI4 and NOD2 delineate risk variant function in rheumatoid arthritis, leprosy and Crohn's disease. Taken together, these data help advance understanding of the genetics of gene expression, neutrophil biology and immune-related diseases.


Reflection and refraction of multipole radiation by an interface.
Reflection and refraction of electromagnetic multipole radiation by an interface is studied. The multipole can be electric or magnetic and is of arbitrary order (dipole, quadrupole). From the angular spectrum representation of the radiation emitted by the multipole, I have obtained the angular spectrum representations of the reflected and transmitted fields, which involve the Fresnel reflection and transmission coefficients. The intensity distribution in the far field is evaluated with the method of stationary phase. The result is very simple in appearance and can be expressed in terms of two auxiliary functions of a complex variable. By exchanging the Fresnel coefficients for s and p polarization, the result for an electric multipole can be obtained from the result for a magnetic multipole.


RosBREED: bridging the chasm between discovery and application to enable DNA-informed breeding in rosaceous crops.
The Rosaceae crop family (including almond, apple, apricot, blackberry, peach, pear, plum, raspberry, rose, strawberry, sweet cherry, and sour cherry) provides vital contributions to human well-being and is economically significant across the U.S. In 2003, industry stakeholder initiatives prioritized the utilization of genomics, genetics, and breeding to develop new cultivars exhibiting both disease resistance and superior horticultural quality. However, rosaceous crop breeders lacked certain knowledge and tools to fully implement DNA-informed breeding-a "chasm" existed between existing genomics and genetic information and the application of this knowledge in breeding. The RosBREED project ("Ros" signifying a Rosaceae genomics, genetics, and breeding community initiative, and "BREED", indicating the core focus on breeding programs), addressed this challenge through a comprehensive and coordinated 10-year effort funded by the USDA-NIFA Specialty Crop Research Initiative. RosBREED was designed to enable the routine application of modern genomics and genetics technologies in U.S. rosaceous crop breeding programs, thereby enhancing their efficiency and effectiveness in delivering cultivars with producer-required disease resistances and market-essential horticultural quality. This review presents a synopsis of the approach, deliverables, and impacts of RosBREED, highlighting synergistic global collaborations and future needs. Enabling technologies and tools developed are described, including genome-wide scanning platforms and DNA diagnostic tests. Examples of DNA-informed breeding use by project participants are presented for all breeding stages, including pre-breeding for disease resistance, parental and seedling selection, and elite selection advancement. The chasm is now bridged, accelerating rosaceous crop genetic improvement.


Localization and production of angiotensin II in the isolated perfused rat heart.
We used a modification of the isolated perfused rat heart, in which coronary effluent and interstitial transudate were separately collected, to investigate the localization and production of angiotensin II (Ang II) in the heart. During combined renin (0.7 to 1.5 pmol Ang I/mL per minute) and angiotensinogen (6 to 12 pmol/mL) perfusion (4 to 8 mL/min) for 60 minutes (n=3), the steady-state levels of Ang II in interstitial transudate in two consecutive 10-minute periods were 4.3+/-1.5 and 3.6+/-1.5 fmol/mL compared with 1.1+/-0.4 and 1.1+/-0.6 fmol/mL in coronary effluent (mean+/-half range). During perfusion with Ang II (n=5), steady-state Ang II in interstitial transudate was 32+/-19% of arterial Ang II compared with 65+/-16% in coronary effluent (mean+/-SD, P<.02). During perfusion with Ang I (n=5), Ang II in interstitial transudate was 5.1+/-0.6% of arterial Ang I compared with 2.2+/-0.3% in coronary effluent (P<.05). The tissue concentration of Ang II in the combined renin/angiotensinogen perfusions (per gram) was as high as the concentration in interstitial transudate (per milliliter). Addition of losartan (10(-6) mol/L) to the renin/angiotensinogen perfusion (n=3) had no significant effect on the tissue level of Ang II, whereas losartan in the perfusions with Ang I (n=5) or Ang II (n=5) decreased tissue Ang II to undetectably low levels. The results indicate that the heart is capable of producing Ang II and that this can lead to higher levels in tissue than in blood plasma. Cardiac Ang II does not appear to be restricted to the extracellular fluid. This is in part due to AT1-receptor-mediated cellular uptake of extracellular Ang II, but our results also raise the possibility of intracellular Ang II production.


Refractory hypertension: determination of prevalence, risk factors, and comorbidities in a large, population-based cohort.
Refractory hypertension is an extreme phenotype of antihypertensive treatment failure. Participants in the REasons for Geographic And Racial Differences in Stroke (REGARDS) Study, a large (n=30 239), population-based cohort were evaluated to determine the prevalence of refractory hypertension and associated cardiovascular risk factors and comorbidities. Refractory hypertension was defined as uncontrolled blood pressure (systolic/diastolic, ≥140/90 mm Hg) on ≥5 antihypertensive drug classes. Participants with resistant hypertension (systolic/diastolic, ≥140/90 mm Hg on ≥3 or <140/90 mm Hg on ≥4 antihypertensive classes) and all participants treated for hypertension served as comparator groups. Of 14 809 REGARDS participants receiving antihypertensive treatment, 78 (0.5%) had refractory hypertension. The prevalence of refractory hypertension was 3.6% among participants with resistant hypertension (n=2144) and 41.7% among participants on ≥5 antihypertensive drug classes. Among all participants with hypertension, black race, male sex, living in the stroke belt or buckle, higher body mass index, lower heart rate, reduced estimated glomerular filtration rate, albuminuria, diabetes mellitus, and history of stroke and coronary heart disease were associated with refractory hypertension. Compared with resistant hypertension, prevalence ratios for refractory hypertension were increased for blacks (3.00; 95% confidence interval, 1.68-5.37) and those with albuminuria (2.22; 95% confidence interval, 1.40-3.52) and diabetes mellitus (2.09; 95% confidence interval, 1.32-3.31). The median 10-year Framingham risk for coronary heart disease and stroke was higher among participants with refractory hypertension when compared with those with either comparator group. These data indicate that although resistant hypertension is relatively common among treated patients with hypertension, true antihypertensive treatment failure is rare.


The versatile biomedical applications of bismuth-based nanoparticles and composites: therapeutic, diagnostic, biosensing, and regenerative properties.
Studies of nanosized forms of bismuth (Bi)-containing materials have recently expanded from optical, chemical, electronic, and engineering fields towards biomedicine, as a result of their safety, cost-effective fabrication processes, large surface area, high stability, and high versatility in terms of shape, size, and porosity. Bi, as a nontoxic and inexpensive diamagnetic heavy metal, has been used for the fabrication of various nanoparticles (NPs) with unique structural, physicochemical, and compositional features to combine various properties, such as a favourably high X-ray attenuation coefficient and near-infrared (NIR) absorbance, excellent light-to-heat conversion efficiency, and a long circulation half-life. These features have rendered bismuth-containing nanoparticles (BiNPs) with desirable performance for combined cancer therapy, photothermal and radiation therapy (RT), multimodal imaging, theranostics, drug delivery, biosensing, and tissue engineering. Bismuth oxyhalides (BiOx, where X is Cl, Br or I) and bismuth chalcogenides, including bismuth oxide, bismuth sulfide, bismuth selenide, and bismuth telluride, have been heavily investigated for therapeutic purposes. The pharmacokinetics of these BiNPs can be easily improved via the facile modification of their surfaces with biocompatible polymers and proteins, resulting in enhanced colloidal stability, extended blood circulation, and reduced toxicity. Desirable antibacterial effects, bone regeneration potential, and tumor growth suppression under NIR laser radiation are the main biomedical research areas involving BiNPs that have opened up a new paradigm for their future clinical translation. This review emphasizes the synthesis and state-of-the-art progress related to the biomedical applications of BiNPs with different structures, sizes, and compositions. Furthermore, a comprehensive discussion focusing on challenges and future opportunities is presented.


Walnut Consumption Alters the Gastrointestinal Microbiota, Microbially Derived Secondary Bile Acids, and Health Markers in Healthy Adults: A Randomized Controlled Trial.
Epidemiologic data suggest that diets rich in nuts have beneficial health effects, including reducing total and cause-specific mortality from cancer and heart disease. Although there is accumulating preclinical evidence that walnuts beneficially affect the gastrointestinal microbiota and gut and metabolic health, these relations have not been investigated in humans. We aimed to assess the impact of walnut consumption on the human gastrointestinal microbiota and metabolic markers of health. A controlled-feeding, randomized crossover study was undertaken in healthy men and women [n = 18; mean age = 53.1 y; body mass index (kg/m2): 28.8]. Study participants received isocaloric diets containing 0 or 42 g walnuts/d for two 3-wk periods, with a 1-wk washout between diet periods. Fecal and blood samples were collected at baseline and at the end of each period to assess secondary outcomes of the study, including effects of walnut consumption on fecal microbiota and bile acids and metabolic markers of health. Compared with after the control period, walnut consumption resulted in a 49-160% higher relative abundance of Faecalibacterium, Clostridium, Dialister, and Roseburia and 16-38% lower relative abundances of Ruminococcus, Dorea, Oscillospira, and Bifidobacterium (P < 0.05). Fecal secondary bile acids, deoxycholic acid and lithocholic acid, were 25% and 45% lower, respectively, after the walnut treatment compared with the control treatment (P < 0.05). Serum LDL cholesterol and the noncholesterol sterol campesterol concentrations were 7% and 6% lower, respectively, after walnut consumption compared with after the control treatment (P < 0.01). Walnut consumption affected the composition and function of the human gastrointestinal microbiota, increasing the relative abundances of Firmicutes species in butyrate-producing Clostridium clusters XIVa and IV, including Faecalibacterium and Roseburia, and reducing microbially derived, proinflammatory secondary bile acids and LDL cholesterol. These results suggest that the gastrointestinal microbiota may contribute to the underlying mechanisms of the beneficial health effects of walnut consumption. This trial was registered at www.clinicaltrials.gov as NCT01832909.


N-butyldeoxynojirimycin-mediated inhibition of human immunodeficiency virus entry correlates with impaired gp120 shedding and gp41 exposure.
The alpha-glucosidase inhibitor N-butyldeoxynojirimycin (NB-DNJ) is an inhibitor of human immunodeficiency virus (HIV) replication and HIV-induced syncytium formation in vitro. Although an NB-DNJ-mediated change in viral envelope N-glycan composition inhibits HIV entry at the level of post-CD4 binding, the exact mechanism of inhibition remains to be established. In this study we have examined the effects of NB-DNJ on virion envelope composition and CD4-induced gp120 shedding and gp41 exposure. Virion composition analysis revealed an NB-DNJ-mediated reduction of 15% in overall virion envelope glycoprotein content and a reduction of 26% in the proteolytic maturation of virion gp160. Taken together, these two effects resulted in a reduction of approximately 40% in virion gp120 content. CD4-induced shedding of gp120 from the surfaces of envelope-transfected Cos cells was undetectable when gp120 was expressed in the presence of NB-DNJ. Similarly, the shedding of virion-associated gp120 was reduced 7.4-fold. CD4-induced exposure of cryptic gp41 epitopes on the surfaces of HIV-expressing ACH-2 cells was also greatly impaired, and the exposure of virion-associated gp41 epitopes was reduced 4.0-fold. Finally, CD4-induced increases in the binding of antibodies to the V3 loop of ACH-2-cell-expressed envelope glycoproteins were reduced 25-fold when the glycoproteins were expressed in the presence of NB-DNJ. These results suggest that the NB-DNJ-mediated retention of glycosylated N-glycans inhibits HIV entry by a combined effect of a reduction in virion gp120 content and a qualitative defect within the remaining gp120, preventing it from undergoing conformational changes after CD4 binding.


An 18-mo randomized, double-blind, placebo-controlled trial of DHA-rich fish oil to prevent age-related cognitive decline in cognitively normal older adults.
Fish oil trials in cognitively healthy older adults have yielded inconsistent results. Supplementation may differentially affect the domains that underpin cognitive performance, and effects may differ across sex or genotype. The aim of this study was to test whether docosahexaenoic acid (DHA)-rich fish oil slows 18-mo cognitive decline in cognitively healthy elders. In a double-blind, randomized, placebo-controlled, parallel-group trial, cognitively healthy Australian community-dwelling adults (aged 65-90 y) consumed either 1720 mg DHA and 600 mg eicosapentaenoic acid or low-polyphenolic olive oil daily, as capsules, for 18 mo. Groups were allocated by permuted-block randomization and stratified by age. Cognitive assessment was conducted at baseline and then every 6 mo. Primary analyses tested the difference between groups in the rate of 18-mo cognitive change via latent growth curve models on any of the following: reasoning, working memory, short-term memory, retrieval fluency, and cognitive speed-related constructs. Treatment interactions with sex and APOE-ε4 were tested. Secondary outcomes were self-reported changes in well-being and everyday functioning, blood pressure, biomarkers of n-3 (ω-3) long-chain polyunsaturated fatty acids (LC PUFAs), lipids, glucose metabolism, inflammation, oxidative stress, DNA damage, and Mini-Mental State Examination. A total of 403 people were randomly assigned. Data from those who completed baseline were analyzed (n = 390; intervention n = 194, control n = 196). Daily supplementation with 2.3 g DHA-rich fish oil for 18 mo did not maintain or improve cognitive performance. A small negative main effect was found on psychomotor speed (intervention = -0.02, 95% CI: -0.04 to 0.00; d = 0.24, P = 0.03). Treatment effects differed according to sex on retrieval fluency and some speed-based domains, including psychomotor speed, and according to APOE-ε4 carrier status on reaction time and reasoning. For secondary outcomes, treatment was associated with increased perceived cognitive mistakes (d = 0.24; P = 0.003), increased oxidative stress, and expected changes in fatty acid metabolism. Findings do not support supplementing older adults with fish oil to prevent cognitive decline. Treatment interactions with sex and APOE-ε4 carrier status warrant further investigation. This trial was registered at the Australia and New Zealand Clinical Trials Register (ANZCTR) as ACTRN12607000278437.


The outer membrane of Treponema pallidum: biological significance and biochemical properties.
Rabbits infected intravenously with Treponema pallidum were not markedly febrile, and the pyrogenicity of treponeme preparations administered intravenously to rabbits was negligible. The antibiotic polymyxin B did not induce any ultrastructural changes on the treponemal surface and was not lethal (immobilizing) for T. pallidum, which was, however, highly susceptible to detergents such as SDS. Extraction of treponemes with Triton X-100 removed the outer membrane (despite the presence of Mg2+) as shown by electron microscopy, and solubilized a limited number of proteins detectable by SDS-PAGE, including a dominant antigen (47 kDal) demonstrated by immunoblotting. None of the proteins were heat-modifiable. Periodic acid-silver staining of polyacrylamide gels for carbohydrate together with protease K digestion did not demonstrate major carbohydrate components in whole treponemes, or in the Triton-soluble fraction. Surface iodination of intact treponemes revealed very little surface exposure of treponemal proteins, although a protein which co-migrated with host albumin was labelled and appeared to be associated with the treponemal surface. Many treponemal proteins were, however, labelled when iodination was done in the presence of Triton. These observations, indicate that the outer membrane of T. pallidum differs significantly from those of many Gram-negative pathogens.


A recovery-oriented approach for an acute psychiatric ward: is it feasible and how does it affect staff satisfaction?
To evaluate professionals' attitudes to recovery and coercion, as well their satisfaction with working conditions before and after the implementation of a recovery-oriented ward concept on an admission ward. Longitudinal study design with two measurement times of the study sample, with a control group assessed at study end. Evaluating the implementation of the recovery concept, attitudes towards recovery, coercion, perceptions of the ward and working satisfaction were assessed with questionnaires and computed using Chi square and ANOVA variance analyses. The members of the intervention ward (n = 17) did not differ from the control group (n = 21), except that control group members were younger. The recovery-orientation of the study ward (ROSE questionnaire) increased significantly (alpha level = 0.05) from study begin to study end (p = 0.003), and compared to the control group (p = 0.002). The attitudes towards coercion did not change significantly in the intervention group, but did so compared to the control group. The contentedness (GMI) and the satisfaction with working conditions (ABB) of the intervention group members compared to control group was significantly higher (GMI: p = 0.004, ABB subscale working conditions: p = 0.043, satisfaction: p = 0.023). The study indicates that recovery-oriented principles can be implemented even in an acute admission ward, increasing team satisfaction with work, while attitudes towards coercion did not change significantly within this single-unit project.


Predicting major neurologic improvement and long-term outcome after thrombolysis using artificial neural networks.
To develop artificial neural network (ANN)-based functional outcome prediction models for patients with acute ischemic stroke (AIS) receiving intravenous thrombolysis based on immediate pretreatment parameters. The derived cohort consisted of 196 patients with AIS treated with intravenous thrombolysis between 2009 and 2017 at Shuang Ho Hospital in Taiwan. We evaluated the predictive value of parameters associated with major neurologic improvement (MNI) at 24 h after thrombolysis as well as the 3-month outcome. ANN models were applied for outcome prediction. The generalizability of the model was assessed through 5-fold cross-validation. The performance of the models was assessed according to the accuracy, sensitivity, specificity, and area under the receiver operating characteristic curve (AUC), RESULTS: The parameters associated with MNI were blood pressure (BP), heart rate, glucose level, consciousness level, National Institutes of Health Stroke Scale (NIHSS) score, and history of diabetes mellitus (DM). The parameters associated with the 3-month outcome were age, consciousness level, BP, glucose level, hemoglobin A1c, history of DM, stroke subtype, and NIHSS score. After adequate training, ANN Model 1 to predict MNI achieved an AUC of 0.944. Accuracy, sensitivity, and specificity were 94.6%, 89.8%, and 95.9%, respectively. ANN Model 2 to predict the 3-month outcome achieved an AUC of 0.933, with accuracy, sensitivity, and specificity of 88.8%, 94.7%, and 86.5%, respectively. The ANN-based models achieved reliable performance to predict MNI and 3-month outcomes after thrombolysis for AIS. The models proposed have clinical value to assist in decision-making, especially when invasive adjuvant strategies are considered.


Lesions of the anteroventral third ventricle and development of stress-induce hypertension in the borderline hypertensive rat.
The anteroventral third ventricle (AV3V) region plays a critical role in the pathogenesis of many forms of experimental hypertension. The present study sought to determine whether the integrity of this area was necessary for the development of stress-induced hypertension in the borderline hypertensive rat (BHR). Male BHRs were assigned to three groups at 8 weeks of age: 1) AV3V lesion, 2) sham lesion, and 3) maturation control. BHRs with AV3V and sham lesions were exposed to 12 weeks of conflict stress (2 hr/day, 5 days/wk). At the end of the conflict protocol period, direct measurement of resting mean arterial pressure indicated that BHRs with sham lesions had significantly higher blood pressure (153 +/- 2.9 mm Hg) than rats with AV3V lesions (126 +/- 5.2 mm Hg) and maturation control rats (133 +/- 4.3 mm Hg). Although AV3V lesions prevented stress-induced hypertension in BHRs, these rats were still capable of transiently raising blood pressure. Specifically, the results also indicate that BHRs with AV3V lesions showed greater increase in blood pressure in response to an electric foot-shock paradigm. This study suggests a critical role for this forebrain region in the production of stress-induced hypertension in genetically predisposed animals.


Zinc-mediated binding of a low-molecular-weight stabilizer of the host anti-viral factor apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3G.
Apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3G (APOBEC3G, A3G), is a human anti-virus restriction protein which works deaminase-dependently and -independently. A3G is known to be ubiquitinated by HIV-1 viral infectivity factor (Vif) protein, leading to proteasomal degradation. A3G contains two zinc ions at the N-terminal domain and the C-terminal domain. Four lysine residues, K(297), K(301), K(303), and K(334), are known to be required for Vif-mediated A3G ubiquitination and degradation. Previously, we reported compound SN-1, a zinc chelator that increases steady-state expression level of A3G in the presence of Vif. In this study, we prepared Biotin-SN-1, a biotinylated derivative of SN-1, to study the SN-1-A3G interaction. A pull-down assay revealed that Biotin-SN-1 bound A3G. A zinc-abstraction experiment indicated that SN-1 binds to the zinc site of A3G. We carried out a SN-1-A3G docking study using molecular operating environment. The calculations revealed that SN-1 binds to the C-terminal domain through Zn(2+), H(216), P(247), C(288), and Y(315). Notably, SN-1-binding covers the H(257), E(259), C(288), and C(291) residues that participate in zinc-mediated deamination, and the ubiquitination regions of A3G. The binding of SN-1 presumably perturbs the secondary structure between C(288) and Y(315), leading to less efficient ubiquitination.


Genetic parameters for stayability, stayability at calving, and stayability at weaning to specified ages for Hereford cows.
Genetic parameters for stayability to six ages (ST1, . . ., ST6), for five measures of stayability to calving (SC2, . . ., SC6), and for five measures of stayability to weaning (SW2, . . ., SW6), were estimated using records of 2,019 Hereford cows collected from 1964 to 1979 from a selection experiment with a control line and three lines selected for weaning weight, yearling weight, and an index of yearling weight and muscle score. The model included birth year of the cow as a fixed effect and the cow's sire as a random effect. Analyses were performed with 1) a generalized linear mixed model for binary data using a probit link with a penalized quasi-likelihood function, and 2) with a linear mixed model using REML. Genetic trends were estimated by regressing weighted means of estimated transmitting abilities (ETA) of sires by birth year of their daughters on birth year. Environmental trends were estimated by regressing solutions for year of birth on birth year. Estimates of heritability (SE) for ST were between 0.09 (0.08) and 0.30 (0.14) for threshold model and between 0.05 (0.04) and 0.19 (0.09) for linear model. Estimates of heritability from linear model analyses transformed to an underlying normal scale were between 0.09 and 0.35. Estimates of heritability (SE) for SC were between 0.29 (0.10) and 0.39 (0.11) and between 0.18 (0.09) and 0.25 (0.08) with threshold and linear models. Estimates of heritability transformed to an underlying normal scale were between 0.30 and 0.40. Estimates of heritability (SE) for SW were between 0.21 (0.14) and 0.47 (0.19) and between 0.12 (0.08) and 0.26 (0.12) with threshold and linear models, respectively. Estimates of heritability transformed to an underlying normal scale were between 0.21 and 0.50. Estimates of genetic and environmental trends for all lines were nearly zero for all traits. Correlations between ETA of sires for stayability to specific ages, for stayability to calving, and for stayability to weaning with threshold and linear models ranged from 0.09 to 0.82, from 0.68 to 0.90, and from 0.67 to 0.87, respectively. Selection for stayability would be possible in a breeding program and could be relatively effective as a result of the moderate estimates of heritability, which would allow selection of sires whose daughters are more likely to remain longer in the herd. Selection for weaning and yearling weights resulted in little correlated response for any of the measures of stayability.


Immunological Fingerprints of Controllers Developing Neutralizing HIV-1 Antibodies.
The induction of broadly neutralizing antibodies (bnAbs) is highly desired for an effective vaccine against HIV-1. Typically, bnAbs develop in patients with high viremia, but they can also evolve in some untreated HIV-1 controllers with low viral loads. Here, we identify a subgroup of neutralizer-controllers characterized by myeloid DCs (mDCs) with a distinct inflammatory signature and a superior ability to prime T follicular helper (Tfh)-like cells in an STAT4-dependent fashion. This distinct immune profile is associated with a higher frequency of Tfh-like cells in peripheral blood (pTfh) and an enrichment for Tfh-defining genes in circulating CD4+ T cells. Correspondingly, monocytes from this neutralizer controller subgroup upregulate genes encoding for chemotaxis and inflammation, and they secrete high levels of IL-12 in response to TLR stimulation. Our results suggest the existence of multi-compartment immune networks between mDCs, Tfh, and monocytes that may facilitate the development of bnAbs in a subgroup of HIV-1 controllers.


Evaluation issues in the Swedish Two-County Trial of breast cancer screening: An historical review.
Objectives To summarize debate and research in the Swedish Two-County Trial of mammographic screening on key issues of trial design, endpoint evaluation, and overdiagnosis, and from these to infer promising directions for the future. Methods A cluster-randomized controlled trial of the offer of breast cancer screening in Sweden, with a single screen of the control group at the end of the screening phase forms the setting for a historical review of investigations and debate on issues of design, analysis, and interpretation of results of the trial. Results There has been considerable commentary on the closure screen of the control group, ascertainment of cause of death, and cluster randomization. The issues raised were researched in detail and the main questions answered in publications between 1989 and 2003. Overdiagnosis issues still remain, but methods of estimation taking full account of lead time and of non-screening influences on incidence (taking place mainly before 2005) suggest that it is a minor phenomenon. Conclusion Despite resolution of issues relating to this trial in peer-reviewed publications dating from years, or even decades ago, issues that already have been addressed continue to be raised. We suggest that it would be more profitable to concentrate efforts on current research issues in breast cancer diagnosis, treatment, and prevention.


PCR-based detection in a micro-fabricated platform.
We present a novel, on-chip system for the electrokinetic capture of bacterial cells and their identification using the polymerase chain reaction (PCR). The system comprises a glass-silicon platform with a set of micro-channels, -chambers, and -electrodes. A platinum thin film resistor, placed in the proximity of the chambers, is used for temperature monitoring. The whole chip assembly is mounted on a Printed Circuit Board (PCB) and wire-bonded to it. The PCB has an embedded heater that is utilized for PCR thermal cycle and is controlled by a Lab-View program. Similar to our previous work, one set of electrodes on the chip inside the bigger chamber (0.6 microl volume) is used for diverting bacterial cells from a flowing stream into to a smaller chamber (0.4 nl volume). A second set of interdigitated electrodes (in smaller chamber) is used to actively trap and concentrate the bacterial cells using dielectrophoresis (DEP). In the presence of the DEP force, with the cells still entrapped in the micro-chamber, PCR mix is injected into the chamber. Subsequently, PCR amplification with SYBR Green detection is used for genetic identification of Listeria monocytogenes V7 cells. The increase in fluorescence is recorded with a photomultiplier tube module mounted over an epifluorescence microscope. This integrated micro-system is capable of genetic amplification and identification of as few as 60 cells of L. monocytogenes V7 in less than 90 min, in 600 nl volume collected from a sample of 10(4) cfu ml(-1). Specificity trials using various concentrations of L. monocytogenes V7, Listeria innocua F4248, and Escherichia coli O157:H7 were carried out successfully using two different primer sets specific for a regulatory gene of L. monocytogenes, prfA and 16S rRNA primer specific for the Listeria spp., and no cross-reactivity was observed.


Targeting early changes in the synovial microenvironment: a new class of immunomodulatory therapy?
Controlled immune responses rely on integrated crosstalk between cells and their microenvironment. We investigated whether targeting proinflammatory signals from the extracellular matrix that persist during pathological inflammation provides a viable strategy to treat rheumatoid arthritis (RA). Monoclonal antibodies recognising the fibrinogen-like globe (FBG) of tenascin-C were generated by phage display. Clones that neutralised FBG activation of toll-like receptor 4 (TLR4), without impacting pathogenic TLR4 activation, were epitope mapped by crystallography. Antibodies stained synovial biopsies of patients at different stages of RA development. Antibody efficacy in preventing RA synovial cell cytokine release, and in modulating collagen-induced arthritis in rats, was assessed. Tenascin-C is expressed early in the development of RA, even before disease diagnosis, with higher levels in the joints of people with synovitis who eventually developed RA than in people whose synovitis spontaneously resolved. Anti-FBG antibodies inhibited cytokine release by RA synovial cells and prevented disease progression and tissue destruction during collagen-induced arthritis. Early changes in the synovial microenvironment contribute to RA progression; blocking proinflammatory signals from the matrix can ameliorate experimental arthritis. These data highlight a new drug class that could offer early, disease-specific immune modulation in RA, without engendering global immune suppression.


Tarsocrural joint polymyxin B concentrations achieved following intravenous regional limb perfusion of the drug via a saphenous vein to healthy standing horses.
To determine whether therapeutic concentrations (> 0.5 to 1.0 μg/mL) of polymyxin B (PB) were achieved in the tarsocrural joint of horses when the drug was administered by IV regional limb perfusion (IV-RLP) via a saphenous vein at doses of 25, 50, and 300 mg and to describe any adverse systemic or local effects associated with such administration. 9 healthy adult horses. In the first of 2 experiments, 6 horses each received 25 and 50 mg of PB by IV-RLP via a saphenous vein with at least 2 weeks between treatments. For each treatment, a tourniquet was placed at the midmetatarsus and another was placed midway between the stifle joint and tarsus. Both tourniquets were removed 30 minutes after the assigned dose was administered. Blood and tarsocrural joint fluid samples were collected for determination of PB concentration before and at predetermined times after drug administration. In experiment 2, 4 horses were administered 300 mg of PB by IV-RLP in 1 randomly selected pelvic limb in a manner identical to that used in experiment 1. For all 3 doses, the mean synovial fluid PB concentration was > 10 times the therapeutic concentration and below the level of quantification at 30 and 1,440 minutes after drug administration, respectively. No adverse systemic or local effects were observed following PB administration. Results suggested that IV-RLP of PB might be a viable alternative for treatment of horses with synovial infections caused by gram-negative bacteria.


Deep sea minerals prolong life span of streptozotocin-induced diabetic rats by compensatory augmentation of the IGF-I-survival signaling and inhibition of apoptosis.
Consumption of deep sea minerals (DSM), such as magnesium, calcium, and potassium, is known to reduce hypercholesterolemia-induced myocardial hypertrophy and cardiac-apoptosis and provide protection against cardiovascular diseases. Heart diseases develop as a lethal complication among diabetic patients usually due to hyperglycemia-induced cardiac-apoptosis that causes severe cardiac-damages, heart failure, and reduced life expectancy. In this study, we investigated the potential of DSM and its related cardio-protection to increase the life expectancy in diabetic rats. In this study, a heart failure rat model was developed by using streptozotocin (65 mg kg(-1) ) IP injection. Different doses of DSM-1× (37 mg kg(-1) day(-1) ), 2× (74 mg kg(-1) day(-1) ) and 3× (111 mg kg(-1) day(-1) ), were administered to the rats through gavages for 4 weeks. The positive effects of DSM on the survival rate of diabetes rats were determined with respect to the corresponding effects of MgSO4 . Further, to understand the mechanism by which DSM enhances the survival of diabetic rats, their potential to regulate cardiac-apoptosis and control cardiac-dysfunction were examined. Echocardiogram, tissue staining, TUNEL assay, and Western blotting assay were used to investigate modulations in the myocardial contractile function and related signaling protein expression. The results showed that DSM regulate apoptosis and complement the cardiomyocyte proliferation by enhancing survival mechanisms. Moreover DSM significantly reduced the mortality rate and enhanced the survival rate of diabetic rats. Experimental results show that DSM administration can be an effective strategy to improve the life expectancy of diabetic subjects by improving cardiac-cell proliferation and by controlling cardiac-apoptosis and associated cardiac-dysfunction. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 769-781, 2016.


Predicting and Tracking Short Term Disease Progression in Amnestic Mild Cognitive Impairment Patients with Prodromal Alzheimer's Disease: Structural Brain Biomarkers.
Early Alzheimer's disease (AD) detection using cerebrospinal fluid (CSF) biomarkers has been recommended as enrichment strategy for trials involving mild cognitive impairment (MCI) patients. To model a prodromal AD trial for identifying MRI structural biomarkers to improve subject selection and to be used as surrogate outcomes of disease progression. APOE ɛ4 specific CSF Aβ42/P-tau cut-offs were used to identify MCI with prodromal AD (Aβ42/P-tau positive) in the WP5-PharmaCog (E-ADNI) cohort. Linear mixed models were performed 1) with baseline structural biomarker, time, and biomarker×time interaction as factors to predict longitudinal changes in ADAS-cog13, 2) with Aβ42/P-tau status, time, and Aβ42/P-tau status×time interaction as factors to explain the longitudinal changes in MRI measures, and 3) to compute sample size estimation for a trial implemented with the selected biomarkers. Only baseline lateral ventricle volume was able to identify a subgroup of prodromal AD patients who declined faster (interaction, p = 0.003). Lateral ventricle volume and medial temporal lobe measures were the biomarkers most sensitive to disease progression (interaction, p≤0.042). Enrichment through ventricular volume reduced the sample size that a clinical trial would require from 13 to 76%, depending on structural outcome variable. The biomarker needing the lowest sample size was the hippocampal subfield GC-ML-DG (granule cells of molecular layer of the dentate gyrus) (n = 82 per arm to demonstrate a 20% atrophy reduction). MRI structural biomarkers can enrich prodromal AD with fast progressors and significantly decrease group size in clinical trials of disease modifying drugs.


Biophysical analysis of Plasmodium falciparum Hsp70-Hsp90 organising protein (PfHop) reveals a monomer that is characterised by folded segments connected by flexible linkers.
Plasmodium falciparum causes the most lethal form of malaria. The cooperation of heat shock protein (Hsp) 70 and 90 is thought to facilitate folding of select group of cellular proteins that are crucial for cyto-protection and development of the parasites. Hsp70 and Hsp90 are brought into a functional complex that allows substrate exchange by stress inducible protein 1 (STI1), also known as Hsp70-Hsp90 organising protein (Hop). P. falciparum Hop (PfHop) co-localises and occurs in complex with the parasite cytosolic chaperones, PfHsp70-1 and PfHsp90. Here, we characterised the structure of recombinant PfHop using synchrotron radiation circular dichroism (SRCD) and small-angle X-ray scattering. Structurally, PfHop is a monomeric, elongated but folded protein, in agreement with its predicted TPR domain structure. Using SRCD, we established that PfHop is unstable at temperatures higher than 40°C. This suggests that PfHop is less stable at elevated temperatures compared to its functional partner, PfHsp70-1, that is reportedly stable at temperatures as high as 80°C. These findings contribute towards our understanding of the role of the Hop-mediated functional partnership between Hsp70 and Hsp90.


The glucoregulatory response to high-intensity aerobic exercise following training in rats with insulin-treated type 1 diabetes mellitus.
An acute bout of exercise elicits a rapid, potentially deleterious, reduction in blood glucose in patients with type 1 diabetes mellitus (T1DM). In the current study, we examined whether a 10-week aerobic training program could alleviate the rapid exercise-associated reduction in blood glucose through changes in the glucoregulatory hormonal response or increased hepatic glycogen storage in an insulin-treated rat model of T1DM. Thirty-two male Sprague-Dawley rats were divided evenly into 4 groups: non-T1DM sedentary (C) (n = 8), non-T1DM exercised (CX) (n = 8), T1DM sedentary (D) (n = 8), and T1DM exercised (DX) (n = 8). Exercise training consisted of treadmill running for 5 days/week (1 h, 27 m/min, 6% grade) for 10 weeks. T1DM was induced by multiple streptozotocin injections (20 mg/kg) followed by implantation of subcutaneous insulin pellets. At week 1, an acute exercise bout led to a significant reduction in blood glucose in DX (p < 0.05), whereas CX exhibited an increase in blood glucose (p < 0.05). During acute exercise, serum epinephrine was increased in both DX and CX (p < 0.05), whereas serum glucagon was increased during recovery only in CX (p < 0.01). Following aerobic training in DX, the exercise-mediated reduction in blood glucose remained; however, serum glucagon increased to the same extent as in CX (p < 0.05). DX exhibited significantly less hepatic glycogen (p < 0.001) despite elevations in glycogenic proteins in the liver (p < 0.05). Elevated serum epinephrine and decreased hepatic adrenergic receptor expression were also evident in DX (p < 0.05). In summary, despite aerobic training in DX, abrupt blood glucose reductions and hepatic glycogen deficiencies were evident. These data suggest that sympathetic overactivity may contribute to deficiencies in hepatic glycogen storage.


Prevention of Congenital Disorders and Care of Affected Children: A Consensus Statement.
As the Sustainable Development Goals are adopted by United Nations member states, children with congenital disorders remain left behind in policies, programs, research, and funding. Although this finding was recognized by the creation and endorsement of the 63rd World Health Assembly Resolution in 2010 calling on United Nations member states to strengthen prevention of congenital disorders and the improvement of care of those affected, there has been little to no action since then. The Sustainable Development Goals call for the global health and development community to focus first and foremost on the most vulnerable and those left behind in the Millennium Development Goal era. To maximize the opportunity for every woman and couple to have a healthy child and to reduce the mortality and severe disability associated with potentially avoidable congenital disorders and their consequences for the children affected, their families and communities, and national health care systems, we propose priority measures that should be taken urgently to address this issue.


Diallyl disulphide, a beneficial component of garlic oil, causes a redistribution of cell-cycle growth phases, induces apoptosis, and enhances butyrate-induced apoptosis in colorectal adenocarcinoma cells (HT-29).
Colon cancer is a leading and expanding cause of death worldwide. A major contributory factor to this disease is diet composition; some components are beneficial (e.g, dietary fiber), whereas others are detrimental (e.g., alcohol). Garlic oil is a prominent dietary constituent that prevents the development of colorectal cancer. This effect is believed to be mainly due to diallyl disulphide (DADS), which selectively induces redox stress in cancerous (rather than normal) cells that leads to apoptotic cell death. However, the detailed mechanism by which DADS causes apoptosis remains unclear. We show that DADS treatment of colonic adenocarcinoma cells (HT-29) initiates a cascade of molecular events characteristic of apoptosis. These include a decrease in cellular proliferation, translocation of phosphatidylserine to the plasma-membrane outer-layer, activation of caspase-3 and -9, genomic DNA fragmentation, and G(2)/M phase cell-cycle arrest. Short-chain fatty acids (SCFAs), particularly butyrate (abundantly produced in the gut by bacterial fermentation of dietary polysaccharides), enhance colonic cell integrity but, in contrast, inhibit colonic cancer cell growth. Combining DADS with butyrate augmented the apoptotic effect of butyrate on HT-29 cells. These results suggest that the anticancerous properties of DADS afford greater benefit when supplied with other favorable dietary factors (short chain fatty acids/polysaccharides) that likewise reduce colonic tumor development.


A homomorphic neural network for modeling and prediction.
A homomorphic feedforward network (HFFN) for nonlinear adaptive filtering is introduced. This is achieved by a two-layer feedforward architecture with an exponential hidden layer and logarithmic preprocessing step. This way, the overall input-output relationship can be seen as a generalized Volterra model, or as a bank of homomorphic filters. Gradient-based learning for this architecture is introduced, together with some practical issues related to the choice of optimal learning parameters and weight initialization. The performance and convergence speed are verified by analysis and extensive simulations. For rigor, the simulations are conducted on artificial and real-life data, and the performances are compared against those obtained by a sigmoidal feedforward network (FFN) with identical topology. The proposed HFFN proved to be a viable alternative to FFNs, especially in the critical case of online learning on small- and medium-scale data sets.


Pseudo-Bayesian Approach for Robust Mode Detection and Extraction Based on the STFT.
This paper addresses the problem of disentangling nonoverlapping multicomponent signals from their observation being possibly contaminated by external additive noise. We aim to extract and to retrieve the elementary components (also called modes) present in an observed nonstationary mixture signal. To this end, we propose a new pseudo-Bayesian algorithm to perform the estimation of the instantaneous frequency of the signal modes from their time-frequency representation. In a second time, a detection algorithm is developed to restrict the time region where each signal component behaves, to enhance quality of the reconstructed signal. We finally deal with the presence of noise in the vicinity of the estimated instantaneous frequency by introducing a new reconstruction approach relying on nonbinary band-pass synthesis filters. We validate our methods by comparing their reconstruction performance to state-of-the-art approaches through several experiments involving both synthetic and real-world data under different experimental conditions.


Herpes simplex virus entry is associated with tyrosine phosphorylation of cellular proteins.
The initial step in herpes simplex virus (HSV) entry is binding of virion glycoprotein (g)C and/or gB to cell surface heparan sulfate. After this initial attachment, gD interacts with cell surface receptor or receptors, and the virion envelope fuses with the cell membrane. Fusion requires viral glycoproteins gB, gD, gL, and gH, but the cellular factors that participate in or the pathways activated by viral entry have not been defined. To determine whether signal transduction pathways are triggered by viral-cell fusion, we examined the association of viral entry with tyrosine phosphorylation of cellular proteins. Using immunoprecipitation and Western blotting, we found that at least three cytoplasmic host cell proteins, designated p80, p104, and p140, become tyrosine phosphorylated within 5-10 min after exposure to HSV-1 or HSV-2. However, no phosphorylation is detected when cells are exposed to a mutant virus deleted in gL that binds but fails to penetrate. Phosphorylation is restored when the gL-deletion virus is grown on a complementing cell line. Viral entry and the phosphorylation of p80, p104, and p140 are inhibited when cells are infected with virus in the presence of protein tyrosine kinase inhibitors. Taken together, these studies suggest that tyrosine phosphorylation of host cellular proteins is triggered by viral entry.


Energy transfer within self-assembled cyclic multichromophoric arrays based on orthogonally arranged donor-acceptor building blocks.
We herein present the coordination-driven supramolecular synthesis and photophysics of a [4+4] and a [2+2] assembly, built up by alternately collocated donor-acceptor chromophoric building blocks based, respectively, on the boron dipyrromethane (Bodipy) and perylene bisimide dye (PBI). In these multichromophoric scaffolds, the intensely absorbing/emitting dipoles of the Bodipy subunit are, by construction, cyclically arranged at the corners and aligned perpendicular to the plane formed by the closed polygonal chain comprising the PBI units. Steady-state and fs time-resolved spectroscopy reveal the presence of efficient energy transfer from the vertices (Bodipys) to the edges (PBIs) of the polygons. Fast excitation energy hopping - leading to a rapid excited state equilibrium among the low energy perylene-bisimide chromophores - is revealed by fluorescence anisotropy decays. The dynamics of electronic excitation energy hopping between the PBI subunits was approximated on the basis of a theoretical model within the framework of Förster energy transfer theory. All energy-transfer processes are quantitatively describable with Förster theory. The influence of structural deformations and orientational fluctuations of the dipoles in certain kinetic schemes is discussed.


M. Gazi Yaşargil: the time in Little Rock.
Microneurosurgery is Professor Yaşargil's legacy. Its impact on patient outcomes, surgeons' abilities, the field of neurosurgery in particular, and the art of surgery in general is great, profound, and everlasting. Professor Yaşargil led a revolution that has transformed neurosurgery into the fine art we practice today. His ingenuity, devotion, energy, and masterful technique have made his contributions so salient that they have earned him recognition as Neurosurgery's Man of the Century. It has been an honor, a great opportunity, and a phenomenal experience to spend the last 5 years with him at the University of Arkansas for Medical Sciences in Little Rock. There, he represents the very best in knowledge, expertise, dexterity, and, above all, devotion to advancing the field of neurosurgery. Most enjoyable have been his stimulating intellect and inspiring vision.


Some issues in predicting patient recruitment in multi-centre clinical trials.
A key paper in modelling patient recruitment in multi-centre clinical trials is that of Anisimov and Fedorov. They assume that the distribution of the number of patients in a given centre in a completed trial follows a Poisson distribution. In a second stage, the unknown parameter is assumed to come from a Gamma distribution. As is well known, the overall Gamma-Poisson mixture is a negative binomial. For forecasting time to completion, however, it is not the frequency domain that is important, but the time domain and that of Anisimov and Fedorov have also illustrated clearly the links between the two and the way in which a negative binomial in one corresponds to a type VI Pearson distribution in the other. They have also shown how one may use this to forecast time to completion in a trial in progress. However, it is not just necessary to forecast time to completion for trials in progress but also for trials that have yet to start. This suggests that what would be useful would be to add a higher level of the hierarchy: over all trials. We present one possible approach to doing this using an orthogonal parameterization of the Gamma distribution with parameters on the real line. The two parameters are modelled separately. This is illustrated using data from 18 trials. We make suggestions as to how this method could be applied in practice.


Symbolic understanding of pictures in low-functioning children with autism: the effects of iconicity and naming.
This research investigated whether symbolic understanding of pictures in low-functioning children with autism is mediated by iconicity and language. In Experiment 1, participants were taught novel words paired with unfamiliar pictures that varied in iconicity (black-and-white line drawings, greyscale photographs, colour line drawings, colour photographs). Unlike mental-age matched typically developing peers, children with autism generally mapped words onto pictures rather than depicted referents, however, they generalised labels more frequently in colour picture conditions. In Experiment 2, children with autism categorised a line drawing with its referent, rather than another picture, regardless of whether it was named. Typically developing children only viewed pictures as symbols when they were labelled. Overall, symbolic understanding of pictures in children with autism is facilitated by iconicity (particularly colour), but not language.


Allium Stents: A Novel Solution for the Management of Upper and Lower Urinary Tract Strictures.
Stents are widely use in endoscopic urological procedures. One of the most important indications is the treatment of urinary tract strictures. Allium™ Medical has introduced several types of stents for the treatment of different types of urinary tract strictures, based on anatomic location. All the stents are made of nitinol and coated with a co-polymer that reduces encrustations. These stents are self-expandable and have a large caliber and a high radial force. They have different shapes, designed especially for the treatment of each type of stricture. One of the most important features of Allium-manufactured stents is the ease of removal, due to their special unraveling feature. The company has introduced the Bulbar Urethral Stent (BUS) for treatment of bulbar urethral strictures; a rounded stent available in different lengths. Initial data on 64 patients with bulbar urethral stricture treated with the BUS showed a significant improvement in symptoms, with minimal complications and few adverse events. For treatment of prostate obstruction in patients unfit for surgery or unwilling to undergo a classical prostatic surgery, the Triangular Prostatic Stent (TPS) was introduced, which has a triangular shape that fits in the prostatic urethra. Its body has a high radial force attached to an anchor (which prevents migration) through a trans-sphincteric wire (which reduces incontinence rate). Initial data on 51 patients showed significant improvement in symptoms and in urinary peak flow rate, with a relatively small number of complications. The Round Posterior Stent (RPS) was designed for treatment of post radical prostatectomy bladder neck contracture. This short, round stent has an anchor, which is placed in the bladder neck. This stent being relatively new, the clinical data are still limited. Ureteral strictures can be treated with the Ureteral Stent (URS), which is round-shaped, available in different lengths, and has an anchor option (for very distal or very proximal strictures). We have previously published data on 107 URSs inserted in patients with ureteral stricture due to several etiologies, including patients who failed previous treatment. All patients were asymptomatic for a long period of follow-up after stent removal, with only one case of re-stenosis. In this paper, we review the urological "covered" stents produced by Allium Medical with the relevant clinical data available at the present time.


Cycloheximide congeners produced by Streptomyces sp. SC0581 and photoinduced interconversion between (E)- and (Z)-2,3-dehydroanhydrocycloheximides.
Three new cycloheximide congeners, 2,3-dehydro-α-epi-isocycloheximide (1), (E)- and (Z)-2,3-dehydroanhydrocycloheximides (2 and 3), together with three known compounds, anhydroisoheximide (4), cycloheximide (5), and isocycloheximide (6), were obtained from the cultures of Streptomyces sp. SC0581. Their structures were elucidated by extensive spectroscopic analysis in combination with theoretical conformational analysis and ECD computations. The photoinduced interconversion between 2 and 3 was observed and verified and the possible reaction path and mechanism were proposed by theoretical computations. The antifungal and cytotoxic activities of 1-6 were evaluated and suggested that 2,3-dehydrogenation results in the loss of the activities and supported that the OH-α is important to the activities of cycloheximide congeners.


Development of a novel intraoral measurement device to determine the biomechanical characteristics of the human periodontal ligament.
Periodontal diseases like gingivitis and periodontitis have damaging effects on the periodontium and commonly affect the mechanical properties of the periodontal ligament (PDL), which in the end might lead to loss of teeth. Monitoring tooth mobility and changes of the material properties of the PDL might help in early diagnosis of periodontal diseases and improve their prognosis. It was the aim of this study to develop a novel intraoral device to determine the biomechanical characteristics of the periodontal ligament. This includes the measurement of applied forces and resulting tooth displacement in order to investigate the biomechanical behaviour of the periodontium with varying loading protocols with respect to velocity and tooth displacement. The developed device uses a piezoelectric actuator to apply a displacement to a tooth's crown, and the resulting force is measured by an integrated force sensor. To measure the tooth displacement independently and non-invasively, two magnets are fixed on the teeth. The change in the magnetic field caused by the movement of the magnets is measured by a total of 16 Hall sensors. The displacement of the tooth is calculated from the movement of the magnets. The device was tested in vitro on premolars of four porcine mandibular segments and in vivo on two volunteers. The teeth were loaded with varying activation curves. Comparing the force progression of different activation velocities, the forces decreased with decreasing velocity. Intensive testing demonstrated that the device fulfils all requirements. After acceptance of the ethical committee, further testing in clinical measurements is planned.


Transfer of photon number statistics from coupling light to stored and retrieved probe light.
We report on the transfer of statistical characteristics from writing and reading coupling light to stored and retrieved probe light pulses in a Λ-type electromagnetically induced transparency (EIT) scheme based on the 5S(1/2)-5P(1/2) transition of (87)Rb atoms. When the coherent probe laser pulse was stored in the EIT medium using the pseudo-thermal coupling light, the characteristics of the pseudo-thermal writing coupling light were transferred to the stored probe laser pulse because of the strong correlation between the probe and coupling light in the EIT medium. The photon number statistics of the retrieved probe light changed from Poisson distribution to Bose-Einstein distribution. Additionally, we measured the change in the properties of the retrieved light pulse due to the photon number statistics of the pseudo-thermal reading coupling light.


Risk factors for unplanned pregnancy among young women in Tanzania.
With effective contraceptives available, unplanned pregnancies are preventable and educational interventions have been cited as a promising platform to increase contraceptive use through improving knowledge. However, results from trials of educational interventions have been disappointing. In order to effectively target future interventions, this study aimed to identify risk factors for unplanned pregnancy among young women in Mwanza, Tanzania. Data were analysed from the MEMA kwa Vijiana Trial Long-term Evaluation Survey, a cross-sectional study of 13,814 young adults aged 15-30 years in Mwanza, Tanzania. Potential risk factors for unplanned pregnancy were grouped under three headings: socio-demographic, knowledge of and attitude towards sexual health, and sexual behaviour and contraceptive use. Conditional logistic regression was used to identify predictors of reported unplanned pregnancy among all sexually active women. Increasing age, lower educational level, not being currently married, knowing where to access condoms, increasing number of sexual partners and younger reported age at sexual debut were associated with unplanned pregnancy. A number of demographic and sexual behaviour risk factors for pregnancy are identified which will help guide future intervention programmes aiming to reduce unplanned pregnancies. This study suggests effective measures to prevent unplanned pregnancies should focus on encouraging girls to stay in school.


Novel morphology-controlled hierarchical core@shell structural organo-layered double hydroxides magnetic nanovehicles for drug release.
Novel hierarchical core@shell structured salicylate (SA) intercalated ZnAl-LDH (layered double hydroxides) magnetic nanovehicles were obtained via a special double-drop coprecipitation strategy assembling organo-ZnAl-LDH nanocrystals onto the surface of Fe3O4 submicrospheres (∼480 nm) from cheap aspirin and Zn- and Al-nitrates in alkaline solutions. The obtained Fe3O4@SA-LDH-r nanovehicles exhibit varied morphologies with hexagonal LDH ab-face horizontal, vertical, and vertical/slant/horizontal to the surfaces of Fe3O4 upon proper mass ratio (r) of Zn-salt to Fe3O4 from 1.93 to 7.71 in a low supersaturation system and possess moderate drug loadings and strong superparamagnetism. An in vitro release study reveals that under "no MF" mode (without external magnetic field) the SA release exhibits the higher accumulated release amount and smaller half-life (t0.5) for firstname.lastname@example.com (41.2%, 1.63 min) and firstname.lastname@example.com (51.1%, 1.66 min) probably owing to their mainly vertical LDH orientations, while the dramatically reduced SA release (10.0%) and greatly elongated t0.5 (25.6 min) for email@example.com may be due to its relatively stronger host-guest interaction and compact horizontally oriented LDH shell stack. Under "MF on" mode, all the magnetic samples show a detectable reduced SA release owing to the particle-particle interactions among the magnetic nanovehicles. The kinetic fittings show that the release processes of all the samples involve the bulk and surface diffusion. The SA release from firstname.lastname@example.com is mainly determined by the interparticle diffusion among the horizontally oriented LDH shell nanocrystals while those of firstname.lastname@example.com and email@example.com mainly involve the interlayer intraparticle diffusion between LDHs layers due to their largely vertical LDH shell nanocrystals.


Compact hybrid TM-pass polarizer for silicon-on-insulator platform.
Hybrid waveguides consisting of a metal plane separated from a high-index medium by a low-index spacer have recently attracted a lot of interest. TM and TE modes are guided in two different layers in these structures and their properties can be controlled in different manners by changing the waveguide dimensions and material properties. We examine the effects of different parameters on the characteristics of the two modes in such structures. We show that by properly choosing the dimensions, it is possible to cut off the TE mode while the TM mode can still be guided in a well-confined manner. Using this property of the hybrid guide, we propose a TM-pass polarizer. The proposed device is very compact and compatible with the silicon-on-insulator platform. Finite-difference time-domain simulation indicates that such a polarizer can provide a high extinction of the TE mode for a reasonable insertion loss of the TM mode.


Heavy metals contamination in water and three species of most consumed fish sampled from Caspian Sea, 2011.
The present study is an attempt to evaluate the heavy metal contamination in the marine environment of the Caspian Sea of Iran. The concentrations of zinc, chromium, cadmium, and lead were measured in water and dorsal muscle of fish sampled from 10 selected stations along Gorgan coast using atomic absorption spectrophotometry during summer of 2011. The average concentrations of lead in water and fish samples (115.29 ± 14.78 and 113.23 ± 27.01 μg/L) were significantly higher than that of cadmium and chromium (P < 0.05). The highest heavy metals content was observed in Cyprinus carpio samples. The heavy metals concentrations in the samples were within the acceptable recommended standards. But, elevating levels of heavy metals in water and fish made a serious concern about ecosystem and food chain contamination. It is recommended that protective authorities should carry out a continual assessment on the levels of pollutants in the sea.


Fine characterization of the Iceman's mtDNA haplogroup.
Starting from specimens of the intestinal contents of the so-called Tyrolean Iceman or Otzi (5,350-5,100 years before present), it was possible by polymerase chain reaction to amplify fragments of the human mitochondrial DNA (mtDNA) control region that correspond to the sequence found in 1994 at the Munich and Oxford laboratories and which had been attributed to the original DNA of the mummy. The particularly favorable condition of the specimens, showing very low contamination levels, made it easier to extend the analyses to the coding region, which had not previously been considered. The mtDNA of the European population is currently divided into nine (H, T, U, V, W, X, I, J, and K) main groups (haplogroups). The K haplogroup, in particular, is composed of two (K1 and K2) subclusters. The results demonstrate that the Iceman's mtDNA belongs to the K1 subcluster, yet it does not fit any of the three known branches (a, b, and c) into which the K1 subcluster is presently divided. In addition, some other sites, reported to be linked to environmental adaptation or pathologies, were investigated.


Developmentally regulated association of plastid division protein FtsZ1 with thylakoid membranes in Arabidopsis thaliana.
FtsZ is a key protein involved in bacterial and organellar division. Bacteria have only one ftsZ gene, while chlorophytes (higher plants and green alga) have two distinct FtsZ gene families, named FtsZ1 and FtsZ2. This raises the question of why chloroplasts in these organisms need distinct FtsZ proteins to divide. In order to unravel new functions associated with FtsZ proteins, we have identified and characterized an Arabidopsis thaliana FtsZ1 loss-of-function mutant. ftsZ1-knockout mutants are impeded in chloroplast division, and division is restored when FtsZ1 is expressed at a low level. FtsZ1-overexpressing plants show a drastic inhibition of chloroplast division. Chloroplast morphology is altered in ftsZ1, with chloroplasts having abnormalities in the thylakoid membrane network. Overexpression of FtsZ1 also induced defects in thylakoid organization with an increased network of twisting thylakoids and larger grana. We show that FtsZ1, in addition to being present in the stroma, is tightly associated with the thylakoid fraction. This association is developmentally regulated since FtsZ1 is found in the thylakoid fraction of young developing plant leaves but not in mature and old plant leaves. Our results suggest that plastid division protein FtsZ1 may have a function during leaf development in thylakoid organization, thus highlighting new functions for green plastid FtsZ.


T cell receptor-independent basal signaling via Erk and Abl kinases suppresses RAG gene expression.
Signal transduction pathways guided by cellular receptors commonly exhibit low-level constitutive signaling in a continuous, ligand-independent manner. The dynamic equilibrium of positive and negative regulators establishes such a tonic signal. Ligand-independent signaling by the precursors of mature antigen receptors regulates development of B and T lymphocytes. Here we describe a basal signal that controls gene expression profiles in the Jurkat T cell line and mouse thymocytes. Using DNA microarrays and Northern blots to analyze unstimulated cells, we demonstrate that expression of a cluster of genes, including RAG-1 and RAG-2, is repressed by constitutive signals requiring the adapter molecules LAT and SLP-76. This TCR-like pathway results in constitutive low-level activity of Erk and Abl kinases. Inhibition of Abl by the drug STI-571 or inhibition of signaling events upstream of Erk increases RAG-1 expression. Our data suggest that physiologic gene expression programs depend upon tonic activity of signaling pathways independent of receptor ligation.


Glucosepane: a poorly understood advanced glycation end product of growing importance for diabetes and its complications.
Advanced glycation end products (AGEs) represent a family of protein, peptide, amino acid, nucleic acid and lipid adducts formed by the reaction of carbonyl compounds derived directly or indirectly from glucose, ascorbic acid and other metabolites such as methylglyoxal. AGE formation in diabetes is of growing importance for their role as markers and potential culprits of diabetic complications, in particular retinopathy, nephropathy and neuropathy. Development of sensitive and specific assays utilizing liquid chromatography mass spectrometry with isotope dilution method has made it possible to detect and quantitate non-UV active AGEs such as carboxymethyl-lysine and glucosepane, the most prevalent AGE and protein crosslink of the extracellular matrix. Below we review studies on AGE formation in two skin biopsies obtained near the closeout of the Diabetes Control and Complications Trial (DCCT), one of which was processed in 2011 for assay of novel AGEs. The results of these analyses show that while several AGEs are associated and predict complication progression, the glucose/fructose-lysine/glucosepane AGE axis is one of the most robust markers for microvascular disease, especially retinopathy, in spite of adjustment for past or future average glycemia. Yet overall little biological and clinical information is available on glucosepane, making this review a call for data in a field of growing importance for diabetes and chronic metabolic diseases of aging.


Are the spatial features of bodily threat limited to the exact location where pain is expected?
Previous research has revealed that anticipating pain at a particular location of the body prioritizes somatosensory input presented there. The present study tested whether the spatial features of bodily threat are limited to the exact location of nociception. Participants judged which one of two tactile stimuli, presented to either hand, had been presented first, while occasionally experiencing a painful stimulus. The distance between the pain and tactile locations was manipulated. In Experiment 1, participants expected pain either proximal to one of the tactile stimuli (on the hand; near condition) or more distant on the same body part (arm; far condition). In Experiment 2, the painful stimulus was expected either proximal to one of the tactile stimuli (hand; near) or on a different body-part at the same body side (leg; far). The results revealed that in the near condition of both experiments, participants became aware of tactile stimuli presented to the "threatened" hand more quickly as compared to the "neutral" hand. Of particular interest, the data in the far conditions showed a similar prioritization effect when pain was expected at a different location of the same body part as well as when pain was expected at a different body part at the same body side. In this study, the encoding of spatial features of bodily threat was not limited to the exact location where pain was anticipated but rather generalized to the entire body part and even to different body parts at the same side of the body.


Free vascularised tissue transfer and brachytherapy for soft-tissue sarcomas of the extremities.
We reviewed the medical files of nine patients with localised soft-tissue sarcomas of the extremities treated with surgical resection, free vascularised tissue transfer, and computerised after-loading iridium-192 high dose rate brachytherapy over a period of 8 years. All patients had external beam radiation therapy in fractions with a total dose of 48-52 Gy, followed by surgery at approximately 4-5 weeks post-irradiation. Surgical treatment included wide resection of the primary tumour in six patients, and marginal resection in three patients. Microsurgical reconstruction of the soft-tissue defect used a free vascularised musculocutaneous flap that included the gracilis flap in five patients, the latissimus dorsi flap in four patients, and the radiovolar flap in one patient. Postoperative brachytherapy dose ranged from 12 to 24 Gy. Adjuvant chemotherapy was administered in five patients with large tumours and marginal resection. At a mean follow-up of 36 months, eight patients were still alive with no evidence of local recurrence or distant metastases; all these patients had good cosmetic and functional outcomes. Two patients died with lung metastases; one of these patients had local tumour recurrence. Wound healing problems and seroma formations were observed in two patients. Intramedullary nailing was required in one patient for a pathological fracture. Tumour resection for soft-tissue sarcomas combined with microsurgical reconstruction of the soft-tissue defect, preoperative external beam radiation and high dose rate brachytherapy results in a high degree of local control with acceptable complications.


ICP0 antagonizes ICP4-dependent silencing of the herpes simplex virus ICP0 gene.
ICP0 is a regulatory protein that plays a critical role in the replication-latency balance of herpes simplex virus (HSV). Absence of ICP0 renders HSV prone to establish quiescent infections, and thus cellular repressor(s) are believed to silence HSV mRNA synthesis when ICP0 fails to accumulate. To date, an ICP0-antagonized repressor has not been identified that restricts HSV mRNA synthesis by more than 2-fold. We report the unexpected discovery that HSV's major transcriptional regulator, ICP4, meets the criteria of a bona fide ICP0-antagonized repressor of viral mRNA synthesis. Our study began when we noted a repressive activity that restricted ICP0 mRNA synthesis by up to 30-fold in the absence of ICP0. When ICP0 accumulated, the repressor only restricted ICP0 mRNA synthesis by 3-fold. ICP4 proved to be necessary and sufficient to repress ICP0 mRNA synthesis, and did so in an ICP4-binding-site-dependent manner. ICP4 co-immunoprecipitated with FLAG-tagged ICP0; thus, a physical interaction likely explains how ICP0 antagonizes ICP4's capacity to silence the ICP0 gene. These findings suggest that ICP0 mRNA synthesis is differentially regulated in HSV-infected cells by the virus-encoded repressor activity embedded in ICP4, and a virus-encoded antirepressor, ICP0. Bacteriophage lambda relies on a similar repression-antirepression regulatory scheme to "decide" whether a given infection will be productive or silent. Therefore, our findings appear to add to the growing list of inexplicable similarities that point to a common evolutionary ancestry between the herpesviruses and tailed bacteriophage.


Epistatic mutations in PUMA BH3 drive an alternate binding mode to potently and selectively inhibit anti-apoptotic Bfl-1.
Overexpression of anti-apoptotic Bcl-2 family proteins contributes to cancer progression and confers resistance to chemotherapy. Small molecules that target Bcl-2 are used in the clinic to treat leukemia, but tight and selective inhibitors are not available for Bcl-2 paralog Bfl-1. Guided by computational analysis, we designed variants of the native BH3 motif PUMA that are > 150-fold selective for Bfl-1 binding. The designed peptides potently trigger disruption of the mitochondrial outer membrane in cells dependent on Bfl-1, but not in cells dependent on other anti-apoptotic homologs. High-resolution crystal structures show that designed peptide FS2 binds Bfl-1 in a shifted geometry, relative to PUMA and other binding partners, due to a set of epistatic mutations. FS2 modified with an electrophile reacts with a cysteine near the peptide-binding groove to augment specificity. Designed Bfl-1 binders provide reagents for cellular profiling and leads for developing enhanced and cell-permeable peptide or small-molecule inhibitors.


Effects of forced movements on learning: Findings from a choice reaction time task in rats.
To investigate how motor sensation facilitates learning, we used a sensory-motor association task to determine whether the sensation induced by forced movements contributes to performance improvements in rats. The rats were trained to respond to a tactile stimulus (an air puff) by releasing a lever pressed by the stimulated (compatible condition) or nonstimulated (incompatible condition) forepaw. When error rates fell below 15%, the compatibility condition was changed (reversal learning). An error trial was followed by a lever activation trial in which a lever on the correct or the incorrect response side was automatically elevated at a preset time of 120, 220, 320, or 420 ms after tactile stimulation. This lever activation induced forepaw movement similar to that in a voluntary lever release response, and also induced body movement that occasionally caused elevation of the other forepaw. The effects of lever activation may have produced a sensation similar to that of voluntary lever release by the forepaw on the nonactivated lever. We found that the performance improvement rate was increased by the lever activation procedure on the incorrect response side (i.e., with the nonactivated lever on the correct response side). Furthermore, the performance improvement rate changed depending on the timing of lever activation: Facilitative effects were largest with lever activation on the incorrect response side at 320 ms after tactile stimulation, whereas hindering effects were largest for lever activation on the correct response side at 220 ms after tactile stimulation. These findings suggest that forced movements, which provide tactile and proprioceptive stimulation, affect sensory-motor associative learning in a time-dependent manner.


Factors associated with DELAY in diagnosis among tuberculosis patients in Hohoe Municipality, Ghana.
Any delay in diagnosis and consequently treatment of TB patients not only increases the infectivity of the disease in the community, but may also lead to more advance disease state, which may result in more complications and expose patients to higher risk of death. The aim of this study was to assess delays in diagnosing new TB patients and the factors associated with these delays in Hohoe Municipality of Ghana. A cross sectional study was carried out among 73 new TB Patients, 15 years or older, registered between 1st June, 2013 and 31st May, 2014 in Hohoe Municipality. Questionnaires were administered to patients to evaluate factors related to delay by patients in seeking care, delays at healthcare facilities, and total diagnostic delay. Logistic regression was used to determine the factors associated with patient delay (>30 days), healthcare services delay (>15 days), and total delay (>45 days). The median total delay was 104 days (inter-quartile range (IQR):17-191). The median patient delay was 59 days (IQR: 5-123), and the median healthcare services delay was 45 days (IQR: 38-128). Not medically insured (AOR = 6.12; 95 % CI: 1.26-29.88; P < 0.025) and perceived stigma (AOR = 5.30; 95 % CI: 1.33-21.18; P < 0.018) were risk factors associated with prolonged patient delay. Multiple healthcare contact following signs and symptoms (AOR = 10.26; 95 %CI: 2.95-35.72; P < 0.0001) was the only risk factor associated with prolonged healthcare services delay. There is a considerable delay in TB case detection mainly due to patients delay in seeking healthcare. The factors associated with patients' delay include lack of medical insurance, perceived stigma, and making multiple healthcare encounters. Health system strengthening towards decentralizing TB diagnosis and management, raising public awareness about the disease, training of healthcare providers, and collaborating with non-formal healthcare providers may reduce long delays in the management of TB.


Medial meniscal extrusion: a validation study comparing different methods of assessment.
Longitudinal cohort studies of knee OA aetiology use MRI to assess meniscal extrusion within the same knee at sequential time points. A validated method of assessment is required to ensure that extrusion is measured at the same location within the knee at each time point. Absolute perpendicular extrusion from the tibial edge can be assessed using the reference standard of segmentation of the tibia and medial meniscus. This is labour intensive and unsuitable for large cohorts. Two methods are commonly used as proxy measurements. Firstly, the apex of the medial tibial spine is used to identify a reproducible MRI coronal slice, from which extrusion is measured. Secondly, the coronal MRI slice of the knee demonstrating the greatest extrusion is used. The purpose of this study was to validate these two methods against the reference standard and to determine the most appropriate method to use in longitudinal cohort studies. We hypothesised that there is no difference in absolute meniscal extrusion measurements between methods. Twenty high-resolution knee MRI scans were obtained in asymptomatic subjects. The tibia and medial meniscus were manually segmented. A custom MATLAB program was used to determine the difference in medial meniscal extrusion of the knee using the reference standard compared to the two other methods. Assessing extrusion using the single coronal MRI slice demonstrating the greatest extrusion overestimates the true extrusion of the medial meniscus. It incorrectly places the greatest meniscal extrusion at the anterior part of the tibia. Assessing extrusion using a consistent anatomical landmark, such as the medial tibial spine, most reliably corresponds to the reference of segmentation and measurement of true perpendicular extrusion from the tibial edge. Clinicians and researchers should consider this when assessing meniscal extrusion in the knee, and how it changes over time. This study suggests measuring meniscal extrusion on the coronal MRI slice corresponding to the apex of the medial tibial spine as this correlates most closely with the true perpendicular extrusion measurements obtained from manually segmented models. Diagnostic, Level I.


Elimination of typhoid: Possibility or pipe dream?
The elimination of typhoid would have a substantial impact on the health of millions of people living in South and Southeast Asia and sub-Saharan Africa. Like many diseases before it, there are significant challenges to elimination, including access to safe drinking water and adequate sanitation practices, the availability of a vaccine, the identification and management of chronic carriers and the emergence of anti-microbial resistance. While provision of clean water and adequate sanitation must remain the ultimate goal, in the short term, required to cornerstone elimination in endemic areas, is an efficacious vaccine. Recent evidence of effectiveness with a tetanus-toxoid conjugate vaccine offers an opportunity for an immunogenic and safe vaccine able to be delivered to young children. Shown to be efficacious in Nepal, where infection is endemic, it may be a viable option in public health programmes moving towards elimination.


Luteinizing hormone in premenopausal women may stimulate uterine leiomyomata development.
Human chorionic gonadotropin (hCG) has proliferative effects on uterine smooth muscle and leiomyoma tissue in vitro. We hypothesized that luteinizing hormone (LH) would have the same effect by activating the LH/hCG receptor, and it would follow that premenopausal women with higher basal LH levels would be more likely to have leiomyomata. Randomly selected women, aged 35 to 49 years, from a prepaid health plan were screened for leiomyomata with pelvic ultrasound. Urine samples collected during the first or last 5 days of the menstrual cycle were analyzed for LH by immunofluorometric assay, and concentrations were corrected for creatinine (n = 523). Logistic regression and Bayes analyses were used to evaluate the association of LH with presence and size of leiomyomata, adjusting for age, and other risk factors. Women with higher LH were more likely to have leiomyomata (adjusted odds ratios for second and third tertiles were 1.7 and 2.0 compared with lower tertile; 95% confidence intervals, 1.0 to 2.7 and 1.2 to 3.4, respectively). The association was stronger for large leiomyomata. Bayes analyses designed to estimate LH effects on tumor onset separately from tumor growth showed significantly accelerated tumor onset but little evidence of effects on tumor growth. Age, an independent risk factor for leiomyomata, was not affected by inclusion of LH in the logistic models. As hypothesized, women with higher LH were more likely to have leiomyomata, but this did not explain the age-related increase in leiomyomata during perimenopausal ages. Determining whether LH is causal or a marker for susceptibility will require further research.


Predictors of outcome after treatment of mild traumatic brain injury: a pilot study.
To determine factors affecting outcome of comprehensive outpatient rehabilitation of individuals who sustained a mild traumatic brain injury. From a 4-year series of referrals, 49 nonconsecutive participants met criteria for mild traumatic brain injury (ie, loss of consciousness <30 minutes, Glasgow Coma Scale score >12). Outpatient, community-based postconcussion clinic at a rehabilitation hospital. Participants and therapy staff completed the Mayo-Portland Adaptability Inventory-Fourth Edition (MPAI-4) at the initiation and conclusion of treatment. Participants were also administered the Trail Making Test at the start of treatment. Participants generally gave poorer adaptability ratings than staff at the beginning and discharge of treatment. Regression analyses revealed that after controlling for baseline ratings, psychiatric history was associated with worse participant-rated MPAI-4 Adjustment scores at treatment discharge, whereas better Trail Making Test Part B performance at initiation of treatment predicted better participant-rated MPAI-4 Ability at treatment discharge. Premorbid demographic and baseline neurocognitive factors should be taken into account prior to comprehensive treatment of mild traumatic brain injury, as they can influence long-term outcomes. Adaptability ratings from both staff and participants can be useful in gaining different perspectives and assessing factors affecting recovery.


Ministry of health clinical practice guidelines: cancer screening.
The Ministry of Health publishes national clinical practice guidelines to provide doctors and patients in Singapore with evidence-based guidance on managing important medical conditions. This article reproduces the introduction and executive summary (with key recommendations from the guidelines) from the Ministry of Health clinical practice guidelines on cancer screening, for the information of readers of the Singapore Medical Journal. Chapters and page numbers mentioned in the reproduced extract refer to the full text of the guidelines, which are available from the Ministry of Health website (http://www.moh.gov. sg/mohcorp/publications.aspx?id=24018). The recommendations should be used with reference to the full text of the guidelines. Following this article are multiple choice questions based on the full text of the guidelines.


Liver transplantation for alcoholic liver disease.
Alcoholic liver disease (ALD) is the major cause of life-threatening liver disease in Western countries. Abstinence from alcohol is the foundation of all treatment programmes for patients with ALD. Liver transplantation is a valuable option for patients with life-threatening ALD. Although the role of liver transplantation in the treatment of alcoholic hepatitis that is unresponsive to medical therapy is controversial, the latest prospective studies support this approach. No single measure gives a reliable estimate of the risk of drinking relapses before or after liver transplantation, but careful evaluation by an addiction specialist with a particular interest in transplant medicine is the best available approach. Survival, both on the waiting list and after the operation, is better in patients with ALD than in patients with HCV infection. Alcohol relapse may lead to liver damage and increased mortality, albeit usually after many years of renewed drinking. After liver transplantation, patients with ALD have increased rates of mortality and morbidity that are attributable to cardiovascular disease and new-onset cancers of the aerodigestive tract. The latter are probably linked to the high prevalence of smoking in this population. Cessation of smoking is thus an important goal in the care of patients with ALD after they have undergone liver transplantation.


Autism, an extreme challenge to integrative medicine. Part 2: medical management.
Autism and allied autistic spectrum disorders (ASD) present myriad behavioral, clinical, and biochemical abnormalities. Parental participation, advanced testing protocols, and eclectic treatment strategies have driven progress toward cure. Behavioral modification and structured education are beneficial but insufficient. Dietary restrictions, including removal of milk and other casein dairy products, wheat and other gluten sources, sugar, chocolate, preservatives, and food coloring are beneficial and prerequisite to benefit from other interventions. Individualized IgG or IgE testing can identify other troublesome foods but not non-immune mediated food sensitivities. Gastrointestinal improvement rests on controlling Candida and other parasites, and using probiotic bacteria and nutrients to correct dysbiosis and decrease gut permeability. Detoxification of mercury and other heavy metals by DMSA/DMPS chelation can have marked benefit. Documented sulfoxidation-sulfation inadequacies call for sulfur-sulfhydryl repletion and other liver p450 support. Many nutrient supplements are beneficial and well tolerated, including dimethylglycine (DMG) and a combination of pyridoxine (vitamin B6) and magnesium, both of which benefit roughly half of ASD cases. Vitamins A, B3, C, and folic acid; the minerals calcium and zinc; cod liver oil; and digestive enzymes, all offer benefit. Secretin, a triggering factor for digestion, is presently under investigation. Immune therapies (pentoxifyllin, intravenous immunoglobulin, transfer factor, and colostrum) benefit selected cases. Long-chain omega-3 fatty acids offer great promise. Current pharmaceuticals fail to benefit the primary symptoms and can have marked adverse effects. Individualized, in-depth clinical and laboratory assessments and integrative parent-physician-scientist cooperation are the keys to successful ASD management.


Design, synthesis and biological evaluation of water soluble and non-aggregated silicon phthalocyanines, naphthalocyanines against A549, SNU-398, SK-MEL128, DU-145, BT-20 and HFC cell lines as potential anticancer agents.
Cancer has become an important public problem in worldwide since cancer incidence and mortality are growing rapidly. In this study, water soluble and non-aggregated silicon (IV) phthalocyanines and naphthalocyanines containing (3,5-bis{3-[3-(diethylamino)phenoxy]propoxy}phenyl)methoxy groups have been synthesized and characterized to investigate their anticancer potential. Their DNA binding/nuclease, topoisomerases inhibition were investigated using UV-Vis absorption, thermal denaturation and agarose gel electrophoresis. The in vitro cytotoxic properties of the compounds on human lung (A549), breast (BT-20), liver (SNU-398), prostate (DU-145), melanoma (SK-Mel 128) carcinoma and human fibroblast (HFC) normal cell lines were evaluated by using MTT assay. In order to determine the mechanism of cancer cell growth suppression, cell cycle analysis was carried out using flow cytometer on A549 cell line. The Kb values of SiPc1a and SiNc2a were 6.85 ± (0.35) × 106 and 1.72 ± (0.16) × 104 M-1 and Tm values of ct-DNA were calculated as 82.02 °C and 78.07 °C, respectively in the presence of both compounds. The ΔTm values of SiPc1a and SiNc2a were calculated as 6.45 and 2.50 °C, respectively. The nuclease effects of SiPc1a and SiNc2a with supercoiled plasmid pBR322 DNA demonstrated that both compounds did not trigger any DNA nuclease effects at the lowest concentrations without irradiation whereas both compounds in the presence of activating agent (H2O2) showed significant plasmid DNA nuclease actions under irradiation (22.5 J/cm2). SiPc1a and SiNc2a inhibited to topoisomerase I on increasing concentrations whilst they had lower inhibition action toward topoisomerase II that of topoisomerase I. The in vitro cytotoxicity studies displayed that SiPc1a had the highest cytotoxic effects among the tested compounds against A549, SNU-398, SK-MEL128, DU-145, BT-20 and HFC cell lines with CC50 values ranged from 0.49 to 2.99 µM. Furthermore, SiPc1a inhibited cell proliferation by cell cycle arrest in G0/G1 phase. All of these results suggested that SiPc1a is a promising candidate as an anticancer agent.