Steady-State Visual-Evoked Potentials as a Biomarker for Concussion: A Pilot Study.
A variety of assessment tools are currently available to help clinicians assess Sports Related Concussion (SRC). Currently, the most widely available tools are neither objective nor portable, and are therefore not ideal for assessment at the site and time of a suspected injury. A portable system was developed to deliver a measurement of the steady-state visual-evoked potential (SSVEP). This system involved a smartphone housed in a Google Cardboard frame, which delivered a 15-Hz flicker visual stimulus while an electroencephalography (EEG) headset recorded EEG signals. Sixty-five rugby union players were tested during their regular season and were stratified into healthy, concussed, and recovered groups based on clinical examination. Their SSVEP response was quantified into a signal-to-noise ratio (SNR). The SNRs of players in each study group were summarized. Additionally, the SNRs of individual players who had baseline, post-injury, and post-recovery readings were analyzed. Sixty-five participants completed a baseline evaluation to measure their SSVEP. Twelve of these participants sustained a medically diagnosed concussion and completed SSVEP re-testing within 72 h. Eight concussed players received follow-up SSVEP testing after recovery. Concussed participants had a lower SNR [2.20 (2.04-2.38)] when compared to their baseline [4.54 (3.79-5.10)]. When clinically recovered, participant SNR was not significantly different to their baseline [4.82 (4.13-5.18)]. The baseline SNRs of the players who experienced a concussion during the season were not different to those of players who did not experience a concussion [4.80 (4.07-5.68)]. This is the first study to identify differences in SSVEP responses in male amateur rugby union players with and without concussion. It is also the first SSVEP demonstration for concussion evaluation at point-of-care. SSVEPs are significantly attenuated in the presence of concussion in these male athletes. Individuals returned to their baseline SSVEP following clinical recovery from the concussive injury. The use of SSVEPs has the potential to be a supplemental aid for the assessment and management of concussion.


Necrotizing fasciitis in a plaster-casted limb: case report.
Casts may be associated with, and mask, serious life-threatening complications, including infection, compartment syndrome, and deep vein thrombosis with or without pulmonary embolism. A 43-year-old woman had necrotizing fasciitis associated with a closed-reduction casting of a tibial fracture. Her treatment highlights the importance of removing a cast and assessing the skin and tissue underneath for signs of infection in patients with suspected infection. Thorough assessment, early diagnosis, and early intervention in necrotizing fasciitis and sepsis are important to improve patients' outcomes.


Contribution of gastroenteropancreatic appetite hormones to protein-induced satiety.
Effects of protein intake on appetite-regulating hormones and their dynamics are unclear. We investigated the satiating effects of meals with varying protein contents and whether there was an effect of dose on appetite-regulating hormones and appetite ratings. Twenty-five men [mean ± SD age: 30.0 ± 8.7 y; body mass index (BMI; in kg/m(2)): 25.9 ± 4.7] participated in the 3-way, randomized, double-blind crossover study. Test meals were isocaloric with 30% of energy from fat and protein content adjusted at the expense of carbohydrate. Test meals were normal protein (NP; 14% of energy from protein), medium-high protein (MHP; 25% of energy from protein), and high protein (HP, 50% of energy from protein). Appetite ratings and blood samples were assessed every 0.5 h for 4 h. An ad libitum lunch was served 4 h after the meal. Protein increased dose-dependently glucagon-like peptide-1 (GLP-1), peptide YY (PYY) 3-36, and glucagon; MHP produced 10%, 7%, and 47% greater responses, respectively; and HP produced 20%, 14%, and 116% greater responses, respectively, than did NP (P < 0.03). Compared with NP, HP increased insulin and cholecystokinin and decreased ghrelin and glucose-dependent insulinotropic polypeptide (P < 0.05). Satiety and fullness dose-dependently increased by 7% and 6% for MHP and 16% and 19% for HP compared with NP (P < 0.001). Hunger and prospective consumption dose-dependently decreased by 15% and 13% for MHP and by 25% and 26% for HP compared with NP (P < 0.0003). There was a combined effect of GLP-1 and PYY 3-36 (P = 0.03) next to the additive effect of GLP-1 (P = 0.006) on the composite appetite score. No difference was shown in ad libitum energy intake. Protein dose-dependently increased satiety and GLP-1, PYY 3-36, and glucagon, which may, at least in part, be responsible for the satiety-stimulating effect of protein. This trial was registered at clinicaltrials.gov as NCT01561235.


Localizing B-Lines in Lung Ultrasonography by Weakly Supervised Deep Learning, In-Vivo Results.
Lung ultrasound (LUS) is nowadays gaining growing attention from both the clinical and technical world. Of particular interest are several imaging-artifacts, e.g., A- and B- line artifacts. While A-lines are a visual pattern which essentially represent a healthy lung surface, B-line artifacts correlate with a wide range of pathological conditions affecting the lung parenchyma. In fact, the appearance of B-lines correlates to an increase in extravascular lung water, interstitial lung diseases, cardiogenic and non-cardiogenic lung edema, interstitial pneumonia and lung contusion. Detection and localization of B-lines in a LUS video are therefore tasks of great clinical interest, with accurate, objective and timely evaluation being critical. This is particularly true in environments such as the emergency units, where timely decision may be crucial. In this work, we present and describe a method aimed at supporting clinicians by automatically detecting and localizing B-lines in an ultrasound scan. To this end, we employ modern deep learning strategies and train a fully convolutional neural network to perform this task on B-mode images of dedicated ultrasound phantoms in-vitro, and on patients in-vivo. An accuracy, sensitivity, specificity, negative and positive predictive value equal to 0.917, 0.915, 0.918, 0.950 and 0.864 were achieved in-vitro, respectively. Using a clinical system in-vivo, these statistics were 0.892, 0.871, 0.930, 0.798 and 0.958, respectively. We moreover calculate neural attention maps that visualize which components in the image triggered the network, thereby offering simultaneous weakly-supervised localization. These promising results confirm the capability of the proposed method to identify and localize the presence of B-lines in clinical lung ultrasonography.


Hydrocarbon-related microbial processes in the deep sediments of the Eastern Mediterranean Levantine Basin.
During the 2011 exploration season of the EV Nautilus in the Mediterranean Sea, we conducted a multidisciplinary study, aimed at exploring the microbial populations below the sediment-water interface (SWI) in the hydrocarbon-rich environments of the Levantine basin. Two c. 1000-m-deep locations were sampled: sediments fueled by methane seepage at the toe of the Palmachim disturbance and a patch of euxinic sediment with high sulfide and methane content offshore Acre, enriched by hydrocarbon from an unknown source. We describe the composition of the microbial population in the top 5 cm of the sediment with 1 cm resolution, accompanied by measurements of methane and sulfate concentrations, and the isotopic composition of this methane and sulfate (δ¹³C(CH₄), δ¹⁸O(SO₄), and δ³⁴S(SO₄)). Our geochemical and microbiological results indicate the presence of the anaerobic methane oxidation (AOM) coupled to bacterial sulfate reduction (BSR). We show that complex methane and sulfur metabolizing microbial populations are present in both locations, although their community structure and metabolic preferences differ due to potential variation in the hydrocarbon source.


Expression of retinoic acid receptor beta in human renal cell carcinomas correlates with sensitivity to the antiproliferative effects of 13-cis-retinoic acid.
The differentiation and growth suppressive effects of retinoic acid are mediated through retinoic acid nuclear receptors (RARs and RXRs), which are ligand-activated transcription factors. Recent data suggest that both altered and regulated expression of RARs are linked to retinoic acid response in a cell context-dependent manner. This study examined the antiproliferative effects of 13-cis-retinoic acid (cRA) on 12 renal cancer cell lines and correlated these findings with the basal and induced expression of RAR-alpha, -beta and -gamma. Eleven of 12 renal cancers that were either resistant to or only minimally inhibited by cRA did not basally express RAR-beta as determined by Northern blot analysis. In these cells, cRA treatment did not induce RAR-beta expression. In contrast, 1 of 12 cell lines (SK-RC-06) was >90% inhibited by cRA and basally expressed RARbeta. Furthermore, RAR-beta mRNA in SK-RC-06 cells was up-regulated by cRA treatment. Amplification of cDNA using PCR and RAR-beta isoform-specific primer pairs revealed that only SK-RC-06 cells expressed the RAR-beta1 isoform. Expression of RAR-alpha transcripts was abundant in all 12 cell lines examined, whereas low levels of RAR-gamma transcripts were detectable in 6 of 10 renal cancers. Expression of RAR-alpha and RAR-gamma was not affected by cRA. These data showing that the majority of renal cancer cell lines are resistant to cRA suggest that: (a) resistance to the antiproliferative action of cRA correlates with repressed RAR-beta mRNA expression; and (b) the antiproliferative effects of cRA in renal cancer cells are mediated through RAR-beta1.


Apoptosis is a major cause of so-called "caseous necrosis" in mycobacterial granulomas in HIV-infected patients.
To demonstrate that so-called "caseous necrosis" is the result of apoptosis and investigate the association of B and T cells, and macrophages with the granulomas and their relationship to some apoptosis-related proteins. Cervical lymph node biopsy specimens from 55 HIV-infected Thai patients with caseating granulomas, confluent caseating granulomas, sarcoid-like granulomas, foamy macrophage response, pseudo-inflammatory tumour response or non-specific lymphoid hyperplasia were examined histologically and for apoptosis by immunostaining for caspase 3 and TUNEL. Classic tuberculoid caseating granulomas in cervical lymph node and lungs from non-HIV-infected patients were also stained with caspase 3. All areas of caseous necrosis frequently displayed extensive apoptosis that readily accounted for the so-called "necrosis". Small foci of apoptosis were present in the other reaction patterns and fibrotic granulomas often showed residual apoptosis. The extent of apoptosis was inversely related to the numbers of identifiable acid-fast bacilli; all epithelioid macrophages revealed strong immunoexpression of the pro-apoptotic proteins Bax and Fas, whereas the anti-apoptotic protein Bcl-2 was not present. Apoptosis occurred in CD68+ macrophages and CD3+ CD8+ T cells; all nodes were deficient of CD4+ cells. CD8+ T cells were intimately related to the apoptotic foci, suggesting a role in the process, particularly in the absence of CD4+ cells. In non-HIV-infected cases, similar extensive apoptosis was confirmed with caspase 3. So-called "caseous necrosis" is shown for the first time to be the result of apoptosis. In the absence of CD4+ cells the findings negate many of the postulated mechanisms of apoptosis in the murine model and have implications for the treatment of mycobacterial infections.


The effect of hypoxia from birth on the regulation of aldosterone in the 7-day-old rat: plasma hormones, steroidogenesis in vitro, and steroidogenic enzyme messenger ribonucleic acid.
Adaptation to hypoxia in the neonate requires an appropriate adrenocortical response. The purpose of this study was to examine the adaptation of the aldosterone pathway in rat pups exposed to hypoxia in vivo from birth to 7 days of age. Neonatal rats (with their lactating dams) were exposed to normoxia (21% O2) or hypoxia (12% O2) continuously for 7 days from birth. Trunk blood was collected, and entire adrenal glands were processed from 7-day-old rats to study the activity of the steroidogenic pathway in dispersed cells and isolated mitochondria, for measurement of expression of the steroidogenic enzyme messenger RNAs (mRNAs) by RT-competitive PCR and in situ hybridization histochemistry, for measurement of zona glomerulosa width by immunohistofluorescent staining for P450c11AS protein, and for measurement of mitochondrial number and distribution by transmission electron microscopy. Exposure to hypoxia for 7 days from birth resulted in a marked increase in plasma ACTH, corticosterone, and aldosterone with no change in PRA. Aldosteronogenesis and P450c11AS activity were both augmented in dispersed cells; this effect was lost in isolated mitochondria (from entire adrenal glands) using a permeable substrate for P450c11AS. There was no significant effect of hypoxia on expression of the steroidogenic enzyme mRNAs measured by RT-competitive PCR or in situ hybridization histochemistry. Finally, hypoxia had no effect on mitochondrial number or stereology as assessed by transmission electron microscopy or on zona glomerulosa width as assessed by staining for P450c11AS protein. We conclude that, as opposed to that in adults, hypoxia in the neonate results in an augmentation of aldosteronogenesis. This effect is not accounted for by a change in steroidogenic enzyme mRNA expression, zona glomerulosa width (i.e. hyperplasia), or mitochondrial number or distribution. This functional augmentation of aldosteronogenesis may be due to a change in mitochondrial permeability to steroid substrates and/or the effect of cytosolic factors that control mitochondrial steroidogenesis.


A multifunctional nanodiamond-based nanoplatform for the enhanced mild-temperature photothermal/chemo combination therapy of triple negative breast cancer via an autophagy regulation strategy.
Owing to its aggressive biological behavior, the lack of specific targets, and the strong therapeutic resistance of triple negative breast cancer (TNBC), current therapeutic strategies are still limited. The combination of multiple treatments has been confirmed as a promising strategy for TNBC therapy. However, the efficacy of combination therapy can be restricted due to increasing therapeutic resistance to various treatments. Herein, we constructed a nanodiamond (ND)-based nanoplatform for augmented mild-temperature photothermal/chemo combination therapy against TNBC, weakening the therapeutic resistance via autophagy inhibition enabled by the NDs. A layer-by-layer self-assembly approach was utilized to construct the ND-based nanoplatform. First, the NDs were modified with protamine sulphate (PS). Meanwhile, the photosensitizer indocyanine green (ICG) and the HSP70 small molecule inhibitor apoptozole (APZ) could be synchronously incorporated to form positively charged PS@ND (ICG + APZ). Then negatively charged hyaluronic acid (HA) was assembled onto the outer face of PS@ND (ICG + APZ) to form the NPIAs. Finally, the positively charged small molecule anti-cancer drug doxorubicin (DOX) could be adsorbed onto the surface of the NPIAs through electrostatic interactions (NPIADs). The resulting NPIADs could be triggered by NIR laser irradiation to exhibit enhanced mild-temperature photothermal therapy (PTT) effects via suppressing the expression of HSP70, and PTT combined with chemotherapy could further enhance the anti-tumor efficacy. Subsequently, the sensitivity of MDA-MB-231 cells could be significantly improved through the weakening of the thermal/drug resistance via autophagy inhibition, leading to augmented combination therapy that is efficient both in vitro and in vivo. Furthermore, the NPIADs could be used as a theranostic nanoplatform for fluorescence (FL) and photoacoustic (PA) imaging. Taken together, this study demonstrated a multifunctional ND-based nanoplatform for FL/PA imaging-guided augmented mild-temperature photothermal/chemo combination therapy via an autophagy regulation strategy against TNBC.


Male vocalization and female choice in the hybridogenetic Rana lessonae/Rana esculenta complex.
In many species, females can improve their fitness by preferring particular males over others. In Palaearctic water frogs of the Rana lessonae/R. esculenta complex the consequences of such mate choice are particularly pronounced. To produce viable offspring, the hybrid R. esculenta (genotype RL) must mate with the parental species R. lessonae (LL); but R. lessonae should avoid mating with R. esculenta, because the resulting hybrid offspring will eliminate the L genome from the germline (hybridogenesis). Hence, there exists a conflict between the sexual parasite (RL) and its sexual host (LL) over the best mating partner. Previous studies have shown a preference for LL males in LL and RL females; but they have also shown that females cannot usually realize their choice when in close proximity to males, because the males forcefully and indiscriminately amplex them. We tested whether females use male vocalizations as a long-distance signal to increase their chances of mating with the preferred LL males. We exposed female R. lessonae and R. esculenta to playbacks of single LL and RL mating calls (experiment 1) and to choruses with a 3:1 excess of LL and RL calls, respectively (experiment 2). In experiment 1, both female types were attracted more by the LL than by the RL calls. In experiment 2, no discrimination between LL- and RL-dominated choruses was observed. The results suggest that females do not use distant male vocalization to approach preferentially ponds or arenas within a pond that hold an excess of LL males. But once they have arrived in a chorus, mating calls from nearby males can direct them to the preferred LL mates. We discuss possible reasons for the failure to discriminate between choruses and the chances for successful choice between individuals within choruses. Copyright 2000 The Association for the Study of Animal Behaviour.


An unusual presentation of adenoid cystic carcinoma of the minor salivary glands with cranial nerve palsy: a case study.
Adenoid Cystic Carcinoma (ACC) is a rare tumor entity and comprises about 1% of all malignant tumor of the oral and maxillofacial region. It is slow growing but a highly invasive cancer with a high recurrence rate. Intracranial ACC is even more infrequent and could be primary or secondary occurring either by direct invasion, hematogenous spread, or perineural spread. We report the first case of the 5th and 6th nerve palsy due to cavernous sinus invasion by adenoid cystic carcinoma. A 49-year-old African American female presented to the emergency room complaining of severe right-sided headache, photophobia, dizziness and nausea, with diplopia. The patient had a 14 year history migraine headaches, hypertension, and mild intermittent asthma. Physical examination revealed right lateral rectus muscle palsy with esotropia. There was numbness in all three divisions of the right trigeminal nerve. Motor and sensory examination of extremities was normal. An MRI of the brain/brain stem was obtained which showed a large mass in the clivus extending to involve the nasopharynx, pterygoid plate, sphenoid and right cavernous sinuses. Biopsy showed an ACC tumor with a cribriform pattern of the minor salivary glands. The patient underwent total gross surgical resection and radiation therapy. This is a case of ACC of the minor salivary glands with intracranial invasion. The patient had long history of headaches which changed in character during the past year, and symptoms of acute 5th and 6th cranial nerve involvement. Our unique case demonstrates direct invasion of cavernous sinus and could explain the 5th and 6th cranial nerve involvement as histopathology revealed no perineural invasion.


Intracellular recordings from neurobiotin-labeled cells in brain slices of the rat medial nucleus of the trapezoid body.
Principal cells in the medial nucleus of the trapezoid body (MNTB) are believed to be critical components in the circuit subserving sound localization. These cells, located in the superior olivary complex, convert excitatory inputs, arriving from the contralateral cochlear nucleus by way of large somatic synapses (the calyces of Held), to inhibitory projections onto principal cells in the ipsilateral lateral superior olive (LSO). We have characterized a population of cells in the rat MNTB using intracellular recording and labeling techniques in a brain slice preparation. MNTB principal cells had spherical or ellipsoid somata that gave rise to single large-diameter dendrites, which branched extensively and often extended beyond the borders of MNTB. Commonly observed axonal projection targets included LSO, the superior paraolivary nucleus, and the medial superior olive, and occasionally the lateral nucleus of the trapezoid body. The projections of individual MNTB cells showed an orderly topography that is consistent with the known tonotopic maps of the nuclei. In response to current injection, principal cells exhibited several nonlinearities, including rectification for depolarizing currents and a "sag" in the membrane potential for hyperpolarizing currents. Superthreshold depolarizing currents elicited transient firing behavior. Application of the potassium channel blocker 4-aminopyridine reduced or eliminated the rectification in the current-voltage relationships and caused depolarizing currents to elicit repetitive firing. Stimulation of afferent inputs elicited short-latency spikes, presumably driven by calyceal synaptic inputs; long-latency, presumably polysynaptic, EPSPs; and short- and long-latency IPSPs. The duration of synaptic events was strongly dependent on membrane potential, and this effect was probably due to the intrinsic membrane properties of the cell. In all cases tested, EPSPs were blocked by CNQX or DNQX, and IPSPs were blocked by strychnine. Two injected non-principal cells differed from principal cells in their morphologies and physiological characteristics.


Associations between null mutations in GSTT1 and GSTM1, the GSTP1 Ile(105)Val polymorphism, and mortality in breast cancer survivors.
Here we assessed associations between null mutations in glutathione-S-transferase (GST)T1 and GSTM1 genes, and the rs1695 polymorphism in GSTP1 (Ile(105)Val), and risk of breast cancer-specific (n=45) and all-cause (n=99) mortality in a multiethnic, prospective cohort of 533 women diagnosed with stage I-IIIA breast cancer in 1995-1999, enrolled in the Health, Eating, Activity, and Lifestyle (HEAL) Study. We measured the presence of the null mutation in GSTT1 and GSTM1, and the rs1695 polymorphism in GSTP1 by polymerase chain reaction. We assessed associations between breast-cancer specific and all-cause mortality using Cox proportional hazards models. Participants with ER-negative tumors were more likely to be GSTT1 null (χ(2)=4.52; P=0.03), and African American women were more likely to be GSTM1 null (χ(2)=34.36; P<0.0001). Neither GSTM1 nor GSTT1 null mutations were associated with breast cancer-specific or all-cause mortality. In a model adjusted for body mass index, race/ethnicity, tumor stage and treatment received at diagnosis, the variant Val allele of rs1695 was associated with increased risk of all-cause (HR=1.81, 95% CI 1.16-2.82, P=0.008), but not breast cancer-specific mortality. The GSTT1 null mutation was associated with significantly higher levels of C-reactive protein. GSTM1 and GSTT1 null genotypes had no effect on outcome; however the variant allele of rs1695 appears to confer increased risk for all-cause mortality in breast-cancer survivors. Given the limited sample size of most studies examining associations between GST polymorphisms with breast cancer survival, and the lack of women undergoing more contemporary treatment protocols (treated prior to 1999), it may be helpful to re-examine this issue among larger samples of women diagnosed after the late 1990s, who all received some form of chemotherapy or radiotherapy.


Sonoprinting liposomes on tumor spheroids by microbubbles and ultrasound.
Ultrasound-triggered drug-loaded microbubbles have great potential for drug delivery due to their ability to locally release drugs and simultaneously enhance their delivery into the target tissue. We have recently shown that upon applying ultrasound, nanoparticle-loaded microbubbles can deposit nanoparticles onto cells grown in 2D monolayers, through a process that we termed "sonoprinting". However, the rigid surfaces on which cell monolayers are typically growing might be a source of acoustic reflections and aspherical microbubble oscillations, which can influence microbubble-cell interactions. In the present study, we aim to reveal whether sonoprinting can also occur in more complex and physiologically relevant tissues, by using free-floating 3D tumor spheroids as a tissue model. We show that both monospheroids (consisting of tumor cells alone) and cospheroids (consisting of tumor cells and fibroblasts, which produce an extracellular matrix) can be sonoprinted. Using doxorubicin-liposome-loaded microbubbles, we show that sonoprinting allows to deposit large amounts of doxorubicin-containing liposomes to the outer cell layers of the spheroids, followed by doxorubicin release into the deeper layers of the spheroids, resulting in a significant reduction in cell viability. Sonoprinting may become an attractive approach to deposit drug patches at the surface of tissues, thereby promoting the delivery of drugs into target tissues.


Mesenchymal cystic hamartoma presenting with spontaneous pneumothorax.
A 53-year-old woman presented with a spontaneous right-sided pneumothorax. Computed tomography chest scan demonstrated a large bulla involving the right middle lobe. Recurrence of the pneumothorax after initial closed chest tube drainage necessitated thoracotomy and wedge resection. Histopathological examination revealed features consistent with a benign mesenchymal cystic hamartoma. Pneumothorax is a recognized complication of mesenchymal cystic hamartoma, a lesion that also has malignant potential.


Short duration combination chemotherapy in the treatment of small cell lung cancer.
Ninety five patients (57 with limited disease and 38 with extensive disease) with previously untreated small cell lung cancer were entered into a study of short duration combination chemotherapy with intravenous cyclophosphamide (750 mg/m2) on day 1, adriamycin (40 mg/m2) on day 1, and etoposide VP-16 (100 mg/m2) on days 1, 2, and 3, with the addition on day 10 of methotrexate 50 mg/m2 with folinic acid rescue and vincristine 2 mg. The treatment was repeated on day 22 and only three courses were given. No maintenance chemotherapy was given, though patients with a complete response received radiotherapy (30-40 Gy (3000-4000 rads] to the primary site in most cases. Forty nine patients (86%) with limited disease achieved a response, with 26 (46%) complete remissions. Twenty five patients (66%) with extensive disease had a response, but only eight (21%) had a complete response. Actuarial survival analysis for the whole patient population showed a median survival of 13 months for patients with limited disease and seven months for those with extensive disease. The median survival was 14 months for those patients with limited disease who achieved a complete response, but only 10 months for non-responders. Myelosuppression was the major expression of toxicity. There were three deaths related to treatment and seven patients had febrile episodes during neutropenia that required antibiotics. Mucositis, which was usually mild, occurred in 49% of patients. The primary site was the main site of initial relapse in 56% of the patients who relapsed. Among patients with limited disease who achieved a complete response, relapses at the primary site were less common in those who received radiotherapy (five out of 12) than in those who did not (all eight). The results indicate that this short duration chemotherapy in small cell lung cancer gives response rates and the potential for long term survival similar to those obtained in other series while allowing patients the maximum time free from treatment.


A comparison of species specific sensitivities to changing light and carbonate chemistry in calcifying marine phytoplankton.
Coccolithophores are unicellular marine phytoplankton and important contributors to global carbon cycling. Most work on coccolithophore sensitivity to climate change has been on the small, abundant bloom-forming species Emiliania huxleyi and Gephyrocapsa oceanica. However, large coccolithophore species can be major contributors to coccolithophore community production even in low abundances. Here we fit an analytical equation, accounting for simultaneous changes in CO2 and light intensity, to rates of photosynthesis, calcification and growth in Scyphosphaera apsteinii. Comparison of responses to G. oceanica and E. huxleyi revealed S. apsteinii is a low-light adapted species and, in contrast, becomes more sensitive to changing environmental conditions when exposed to unfavourable CO2 or light. Additionally, all three species decreased their light requirement for optimal growth as CO2 levels increased. Our analysis suggests that this is driven by a drop in maximum rates and, in G. oceanica, increased substrate uptake efficiency. Increasing light intensity resulted in a higher proportion of muroliths (plate-shaped) to lopadoliths (vase shaped) and liths became richer in calcium carbonate as calcification rates increased. Light and CO2 driven changes in response sensitivity and maximum rates are likely to considerably alter coccolithophore community structure and productivity under future climate conditions.


snow white, a zebrafish model of Hermansky-Pudlak Syndrome type 5.
Hermansky-Pudlak Syndrome (HPS) is a set of genetically heterogeneous diseases caused by mutations in one of nine known HPS genes. HPS patients display oculocutaneous hypopigmentation and bleeding diathesis and, depending on the disease subtype, pulmonary fibrosis, congenital nystagmus, reduced visual acuity, and platelet aggregation deficiency. Mouse models for all known HPS subtypes have contributed greatly to our understanding of the disease, but many of the molecular and cellular mechanisms underlying HPS remain unknown. Here, we characterize ocular defects in the zebrafish (Danio rerio) mutant snow white (snw), which possesses a recessive, missense mutation in hps5 (hps5I76N). Melanosome biogenesis is disrupted in snw/hps5 mutants, resulting in hypopigmentation, a significant decrease in the number, size, and maturity of melanosomes, and the presence of ectopic multi-melanosome clusters throughout the mutant retina and choroid. snw/hps5I76N is the first Hps5 mutation identified within the N-terminal WD40 repeat protein-protein binding domain. Through in vitro coexpression assays, we demonstrate that Hps5I76N retains the ability to bind its protein complex partners, Hps3 and Hps6. Furthermore, while Hps5 and Hps6 stabilize each other's expression, this stabilization is disrupted by Hps5I76N. The snw/hps5I76N mutant provides a valuable resource for structure-function analyses of Hps5 and enables further elucidation of the molecular and cellular mechanisms underlying HPS.


Effects of coffee and caffeine anhydrous on strength and sprint performance.
Caffeine and coffee are widely used among active individuals to enhance performance. The purpose of the current study was to compare the effects of acute coffee (COF) and caffeine anhydrous (CAF) intake on strength and sprint performance. Fifty-four resistance-trained males completed strength testing, consisting of one-rep max (1RM) and repetitions to fatigue (RTF) at 80% of 1RM for leg press (LP) and bench press (BP). Participants then completed five, 10-second cycle ergometer sprints separated by one minute of rest. Peak power (PP) and total work (TW) were recorded for each sprint. At least 48 hours later, participants returned and ingested a beverage containing CAF (300 mg flat dose; yielding 3-5 mg/kg bodyweight), COF (8.9 g; 303 mg caffeine), or placebo (PLA; 3.8 g non-caloric flavouring) 30 minutes before testing. LP 1RM was improved more by COF than CAF (p = .04), but not PLA (p = .99). Significant interactions were not observed for BP 1RM, BP RTF, or LP RTF (p > .05). There were no sprint × treatment interactions for PP or TW (p > .05). 95% confidence intervals revealed a significant improvement in sprint 1 TW for CAF, but not COF or PLA. For PLA, significant reductions were observed in sprint 4 PP, sprint 2 TW, sprint 4 TW, and average TW; significant reductions were not observed with CAF or COF. Neither COF nor CAF improved strength outcomes more than PLA, while both groups attenuated sprint power reductions to a similar degree. Coffee and caffeine anhydrous may be considered suitable pre-exercise caffeine sources for high-intensity exercise.


Optically addressed ferroelectric memory with nondestructive readout.
We present a review of the emerging optically addressed ferroelectric memory with nondestructive readout as a nonvolatile memory technology, identify its high-impact applications, and project on some novel device designs and architectures that will enable its realization. Based on the high-speed bidirectional polarization-dependent photoresponse, simulation of a readout circuit for a 16-kbit VLSI ferromemory chip yields read-access times of ~20 ns and read-cycle times of ~30 ns (~34 ns and ~44 ns, respectively, within a framework of a radiation-hard environment), easily surpassing those of the conventional electrical destructive readout. Extension of the simulation for a 64-kbit memory shows that the read-access and -cycle times are only marginally increased to ~21 ns and ~31 ns, respectively (~38 ns and ~48 ns, with a radiation-hard readout circuitry). Commercial realization of the optical nondestructive readout, however, would require a reduction in the incident (optical) power by roughly an order of magnitude for the readout or an enhancement in the delivered power-to-size ratio of semiconductor lasers for compact implementation. We present a new two-capacitor memory-cell configuration that provides an enhanced bipolar optoelectronic response from the edges of the capacitor at incident power as low as ~ 2 mW/µm(2). A novel device design based on lead zirconate titanate with the c axis parallel to the substrate is suggested to reduce the requirement of incident optical power further by orders of magnitude.


Randomized, double-blind trial of long-chain polyunsaturated fatty acid supplementation with fish oil and borage oil in preterm infants.
To test the efficacy and safety of long-chain polyunsaturated fatty acid (LCPUFA) supplementation with gamma-linolenic acid, a precursor of arachidonic acid, and docosahexaenoic acid in preterm infants. Preterm (<35 weeks, < or =2000 g birth weight) infants (n=238) randomly assigned to unsupplemented or LCPUFA-supplemented formula to 9 months after term. The main outcome measure was the Bayley Mental and Psychomotor Indexes (MDI, PDI) at 18 months after term. Safety outcome measures were anthropometry (9 and 18 months), feed tolerance, infection, and clinical complications. There were no significant differences in neurodevelopment between groups overall. In preplanned subgroup analyses, LCPUFA-supplemented boys had significantly higher Bayley MDI than did control boys (difference, 5.7 points; 95% CI, 0.3 to 11.1; P=.04). LCPUFA-supplemented infants showed significantly greater weight gain (difference, 310 g; 95% CI, 30 to 590 g; P=.03) and length gain (difference, 1.0 cm; 95% CI, 0.02 to 1.9; P=.05) between birth and 9 months, with greater effect in boys (weight difference at 9 months, 510 g; 95% CI, 80 to 930 g; P=.02; length difference at 18 months, 1.8 cm; 95% CI, 0.1 to 1.8; P=.03). This trial, using the strategy of providing gamma-linolenic acid as a source of arachidonic acid, showed efficacy for growth and for neurodevelopment in boys, with no adverse effects. These data have important implications for LCPUFA-supplementation strategy in preterm infants.


Isolated new onset 'atypical' optic neuritis in the NMO clinic: serum antibodies, prognoses and diagnoses at follow-up.
Severe, recurrent or bilateral optic neuritis (ON) often falls within the neuromyelitis optica spectrum disorders (NMOSD), but the diagnosis can be particularly challenging and has important treatment implications. We report the features, course and outcomes of patients presenting with atypical ON when isolated at onset. We retrospectively analyzed 69 sequential patients referred to a single UK NMO center with isolated ON at onset. Aquaporin-4 antibody (AQP4-Ab) assessment was performed in all patients and IgG1 myelin-oligodenrocyte glycoprotein (MOG-Ab) in AQP4-Ab(neg) patients. 37 AQP4-Ab positive (AQP4-Ab(pos)) and 32 AQP4-Ab negative (AQP4-Ab(neg)) patients (8 with MOG-Ab) were identified. The AQP4-Ab(neg) group included heterogeneous diagnoses: multiple sclerosis (MS), NMO, relapsing isolated ON (RION), monophasic isolated ON and relapsing acute disseminated encephalomyelitis (ADEM)-like syndromes. Compared to AQP4-Ab(neg) patients, AQP4-Ab(pos) patients had a worse residual visual outcome from first attack (median VFSS 4 vs. 0, p = 0.010) and at last assessment (median VFSS 5 versus 2, p = 0.005). However, AQP4-Ab(neg) patients with RION also had poor visual outcome. Up to 35% of AQP4-Ab(neg) patients developed a LETM and two developed low positivity for AQP4-Ab over time. Eight AQP4-Ab(neg) patients (25%) were MOG-Ab positive, covering a range of phenotypes excluding MS; the first ON attack was often bilateral and most had relapsing disease with a poor final visual outcome [VFSS 4, range (0-6)]. In conlcusion, AQP4-Ab positivity is confirmed as a predictor of poor visual outcome but AQP4-Ab(neg) RION also had a poor visual outcome. Of those without AQP4-Ab, 25% had MOG-Ab and another 25% developed MS; thus, MOG-Ab is associated with AQP4-Ab(neg) non-MS ON.


Electrocardiography Screening for Hypertrophic Cardiomyopathy.
Hypertrophic cardiomyopathy (HCM) is the leading cause of sudden cardiac death in athletes. However, preparticipation electrocardiogram (ECG) screening has been criticized for failing to meet cost-effectiveness thresholds, in part because of high false-positive rates. We sought to evaluate whether a highly automated software algorithm could be used for a high throughput, population-based screening program and address several of the limitations seen with population-based screening A proprietary computed algorithm was created based on both voltage- as well as Seattle-based ECG criteria. Different cut points for Q-wave depth, Q-wave length, the degree of ST depression, the degree of T-wave inversion, and left ventricular voltage were analyzed for optimum sensitivity and specificity. After developing receiver operating characteristic curves for each criterion, different cut points were trialed together on our data set to obtain settings to optimize sensitivity and specificity. The automated algorithm was capable of identifying patients with HCM based on ECG with 88.6% sensitivity and 98% specificity, compared to a sensitivity of 90.2% and specificity of 96% when the ECGs were read by physicians according to the Seattle Criteria. Adding voltage criteria improved the sensitivity of the algorithm with a mild decrease in specificity. Optimum sensitivity with this automated software was 98%; optimum specificity was 96%. Computer-automated ECG screening for HCM is feasible. Evaluation of automated ECG algorithms in larger and more diverse populations is warranted.


Evaluation of a safe and sensitive Spike protein-based immunofluorescence assay for the detection of antibody responses to SARS-CoV.
Previously, we have identified a truncated antigenic fragment named protein C [441 to 700 amino acids (a.a.)] as the immunodominant fragment of Spike (S) protein of severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV). We have now successfully expressed protein C using the baculovirus system in S. frugiperda (Sf-9) cells. This recombinant baculovirus expressing protein C was first characterized using five SARS convalescent human sera and five normal human sera. The results showed that protein C is an authentic antigen against SARS-CoV antibody. Our Spike protein-based immunoflourescence assay (IFA) based on this recombinant baculovirus-Sf-9 system was further assessed with a panel of 163 clinical samples collected during the SARS epidemic in Singapore, which include samples from 21 clinically confirmed SARS, 42 non-SARS patient sera, and 100 normal sera. The results were compared to a commercial SARS IFA kit (EUROIMMUN, Germany) and a conventional IFA test performed in Singapore General Hospital. All of the 21 SARS-positive serum samples could be recognized by our IFA, giving a specificity and sensitivity of 100%, which was compatible with both whole virus-based IFA assays. No cross-reactivity with serum samples against infectious bronchitis virus (IBV) and transmissible gastroenteritis virus (TGEV) were detected in our assays. Thus, our Spike protein-based IFA could offer a safer procedure which can be performed in a BSL-2 laboratory as it could mimic the whole virus based-IFA without any loss of sensitivity and specificity. It is also more user-friendly and cost-effective than the whole virus-based IFA.


Perception-action learning as an epistemologically-consistent model for self-updating cognitive representation.
As well as having the ability to formulate models of the world capable of experimental falsification, it is evident that human cognitive capability embraces some degree of representational plasticity, having the scope (at least in infancy) to modify the primitives in terms of which the world is delineated. We hence employ the term 'cognitive bootstrapping' to refer to the autonomous updating of an embodied agent's perceptual framework in response to the perceived requirements of the environment in such a way as to retain the ability to refine the environment model in a consistent fashion across perceptual changes.We will thus argue that the concept of cognitive bootstrapping is epistemically ill-founded unless there exists an a priori percept/motor interrelation capable of maintaining an empirical distinction between the various possibilities of perceptual categorization and the inherent uncertainties of environment modeling.As an instantiation of this idea, we shall specify a very general, logically-inductive model of perception-action learning capable of compact re-parameterization of the percept space. In consequence of the a priori percept/action coupling, the novel perceptual state transitions so generated always exist in bijective correlation with a set of novel action states, giving rise to the required empirical validation criterion for perceptual inferences. Environmental description is correspondingly accomplished in terms of progressively higher-level affordance conjectures which are likewise validated by exploratory action.Application of this mechanism within simulated perception-action environments indicates that, as well as significantly reducing the size and specificity of the a priori perceptual parameter-space, the method can significantly reduce the number of iterations required for accurate convergence of the world-model. It does so by virtue of the active learning characteristics implicit in the notion of cognitive bootstrapping.


How Severe Anaemia Might Influence the Risk of Invasive Bacterial Infections in African Children.
Severe anaemia and invasive bacterial infections are common causes of childhood sickness and death in sub-Saharan Africa. Accumulating evidence suggests that severely anaemic African children may have a higher risk of invasive bacterial infections. However, the mechanisms underlying this association remain poorly described. Severe anaemia is characterized by increased haemolysis, erythropoietic drive, gut permeability, and disruption of immune regulatory systems. These pathways are associated with dysregulation of iron homeostasis, including the downregulation of the hepatic hormone hepcidin. Increased haemolysis and low hepcidin levels potentially increase plasma, tissue and intracellular iron levels. Pathogenic bacteria require iron and/or haem to proliferate and have evolved numerous strategies to acquire labile and protein-bound iron/haem. In this review, we discuss how severe anaemia may mediate the risk of invasive bacterial infections through dysregulation of hepcidin and/or iron homeostasis, and potential studies that could be conducted to test this hypothesis.


Radiological features of hepatocellular carcinoma in Southern Pakistan.
We describe the radiological features of 201 patients diagnosed as having hepatocellular carcinoma (HCC) in Southern Pakistan. The cases of biopsy-proven HCC were analysed retrospectively for years 1994-1998. Age, sex, underlying cirrhosis, hepatitis markers, and radiological description were recorded. The mean age was 56 years. There were 149 males and 52 females. 82% patients had underlying cirrhosis. The tumour size was measured in at least two dimensions, and the maximum mean tumour size at the time of diagnosis was 8.3 +/- 4.0 cm. Of the tumours 79.5% were more than 5 cm; 56% of primary HCC were multifocal; 51% involved the right lobe only; 15% involved the left lobe; 34% involved both lobes of liver. Portal vein thrombosis was detectable in 17%. There were no significant differences in the radiological patterns amongst patients who had hepatitis B virus related and hepatitis C virus-related HCC except for age.


Secondary transfer effects of intergroup contact via social identity complexity.
Secondary transfer effects (STEs) of intergroup contact refer to the generalization of contact effects from a primary encountered outgroup to attitudes towards secondary outgroups (Pettigrew, 2009). Using two large, cross-sectional data sets from Germany (N = 1,381) and Northern Ireland (N = 1,948), this article examined the extent to which STEs of intergroup contact on attitudes towards a range of secondary outgroups occur via a previously unexplored psychological construct, social identity complexity (operationalized as similarity complexity and overlap complexity). Study 1 found primary outgroup contact to be associated with greater similarity complexity, but no indirect effects on secondary outgroup attitudes via complexity emerged. Study 2, however, revealed indirect positive relationships between primary outgroup contact and secondary outgroup attitudes via increased similarity complexity and overlap complexity. These relationships were obtained while controlling for two previously tested mediating mechanisms, attitude generalization (operationalized as primary outgroup attitude) and deprovincialization (operationalized as ingroup attitude and identification). We discuss the theoretical implications of these findings and the contribution of social identity complexity to understanding processes underlying STEs of contact.


Evidence update on prevention of surgical site infection.
Surgical site infection (SSI) is a common healthcare-associated infection and complicates up to 10-20% of operations with considerable strain on healthcare resources. Apart from the widely adopted use of appropriate hair removal, antibiotic prophylaxis, avoidance of hypothermia and perioperative glycaemic control to reduce SSIs, this review has considered new research and systematic reviews, and whether their findings should be included in guidelines. The efficacy of preoperative bathing/showering, antibiotic prophylaxis for clean surgery and perioperative oxygen supplementation to reduce the risk of SSI is still in doubt. By contrast, the use of 2% chlorhexidine in alcohol skin preparation, postoperative negative pressure wound therapy and antiseptic surgical dressings do show promise. Antimicrobial sutures in independent meta-analyses were found to reduce the risk of SSI after all classes of surgery (except dirty) whereas the use of wound guards, or diathermy skin incision (compared with scalpel incision), did not. The incidence of SSI after surgery is not falling. Based on this review of published trials and evidence-based systematic reviews some advances might be included into these care bundles. More research is needed together with improved compliance with care bundles.


Identification of a new family of enzymes with potential O-acetylpeptidoglycan esterase activity in both Gram-positive and Gram-negative bacteria.
The metabolism of the rigid bacterial cell wall heteropolymer peptidoglycan is a dynamic process requiring continuous biosynthesis and maintenance involving the coordination of both lytic and synthetic enzymes. The O-acetylation of peptidoglycan has been proposed to provide one level of control on these activities as this modification inhibits the action of the major endogenous lytic enzymes, the lytic transglycosylases. The O-acetylation of peptidoglycan also inhibits the activity of the lysozymes which serve as the first line of defense of host cells against the invasion of bacterial pathogens. Despite this central importance, there is a dearth of information regarding peptidoglycan O-acetylation and nothing has previously been reported on its de-acetylation. Homology searches of the genome databases have permitted this first report on the identification of a potential family of O-Acetylpeptidoglycan esterases (Ape). These proteins encoded in the genomes of a variety of both Gram-negative and Gram-positive bacteria, including a number of important human pathogens such as species of Neisseria, Helicobacter, Campylobacter, and Bacillus anthracis, have been organized into three families based on amino acid sequence similarities with family 1 being further divided into three sub-families. The genes encoding these proteins are shown to be clustered with Peptidoglycan O-acetyltransferases (Pat) and in some cases, together with other genes involved in cell wall metabolism. Representative bacteria that encode the Ape proteins were experimentally shown to produce O-acetylated peptidoglycan. The hypothetical proteins encoded by the pat and ape genes have been organized into families based on sequence similarities. The Pat proteins have sequence similarity to Pseudomonas aeruginosa AlgI, an integral membrane protein known to participate in the O-acetylation of the exopolysaccaride, alginate. As none of the bacteria that harbor the pat genes produce alginate, we propose that the Pat proteins serve to O-acetylate peptidoglycan which is known to be a maturation event occurring in the periplasm. The Ape sequences have amino acid sequence similarity to the CAZy CE 3 carbohydrate esterases, a family previously known to be composed of only O-acetylxylan esterases. They are predicted to contain the alpha/beta hydrolase fold associated with the GDSL and TesA hydrolases and they possess the signature motifs associated with the catalytic residues of the CE3 esterases. Specific signature sequence motifs were identified for the Ape proteins which led to their organization into distinct families. We propose that by expressing both Pat and Ape enzymes, bacteria would be able to obtain a high level of localized control over the degradation of peptidoglycan through the attachment and removal of O-linked acetate. This would facilitate the efficient insertion of pores and flagella, localize spore formation, and control the level of general peptidoglycan turnover.


Can Laws Be a Potential PET Image Texture Analysis Approach for Evaluation of Tumor Heterogeneity and Histopathological Characteristics in NSCLC?
We investigated the association between the textural features obtained from 18F-FDG images, metabolic parameters (SUVmax, SUVmean, MTV, TLG), and tumor histopathological characteristics (stage and Ki-67 proliferation index) in non-small cell lung cancer (NSCLC). The FDG-PET images of 67 patients with NSCLC were evaluated. MATLAB technical computing language was employed in the extraction of 137 features by using first order statistics (FOS), gray-level co-occurrence matrix (GLCM), gray-level run length matrix (GLRLM), and Laws' texture filters. Textural features and metabolic parameters were statistically analyzed in terms of good discrimination power between tumor stages, and selected features/parameters were used in the automatic classification by k-nearest neighbors (k-NN) and support vector machines (SVM). We showed that one textural feature (gray-level nonuniformity, GLN) obtained using GLRLM approach and nine textural features using Laws' approach were successful in discriminating all tumor stages, unlike metabolic parameters. There were significant correlations between Ki-67 index and some of the textural features computed using Laws' method (r = 0.6, p = 0.013). In terms of automatic classification of tumor stage, the accuracy was approximately 84% with k-NN classifier (k = 3) and SVM, using selected five features. Texture analysis of FDG-PET images has a potential to be an objective tool to assess tumor histopathological characteristics. The textural features obtained using Laws' approach could be useful in the discrimination of tumor stage.


Next generation sequencing data of a defined microbial mock community.
Generating sequence data of a defined community composed of organisms with complete reference genomes is indispensable for the benchmarking of new genome sequence analysis methods, including assembly and binning tools. Moreover the validation of new sequencing library protocols and platforms to assess critical components such as sequencing errors and biases relies on such datasets. We here report the next generation metagenomic sequence data of a defined mock community (Mock Bacteria ARchaea Community; MBARC-26), composed of 23 bacterial and 3 archaeal strains with finished genomes. These strains span 10 phyla and 14 classes, a range of GC contents, genome sizes, repeat content and encompass a diverse abundance profile. Short read Illumina and long-read PacBio SMRT sequences of this mock community are described. These data represent a valuable resource for the scientific community, enabling extensive benchmarking and comparative evaluation of bioinformatics tools without the need to simulate data. As such, these data can aid in improving our current sequence data analysis toolkit and spur interest in the development of new tools.


Variants implicated in cortisone reductase deficiency do not contribute to susceptibility to common forms of polycystic ovary syndrome.
There are close phenotypic similarities between cortisone reductase deficiency (CRD), a rare abnormality of cortisone metabolism, and polycystic ovary syndrome (PCOS). As there is evidence that CRD results from digenic mutations involving the genes encoding 11beta-hydroxysteroid dehydrogenase type 1 (HSD11B1) and hexose-6-phosphate dehydrogenase (H6PD), we sought to establish whether CRD-associated variants in these genes, individually or in combination, influence susceptibility to PCOS. Case-control, family-based association and quantitative-trait analyses. A UK case sample comprising 256 nuclear families ascertained from a PCOS offspring and 213 singleton PCOS cases plus 549 control subjects. All subjects were genotyped for CRD-related variants in HSD11B1 (rs12086634) and H6PD (rs6688832). Testosterone was measured with an in-house radioimmunoassay using ether extraction and dextran-coated charcoal separation. Case-control analyses revealed no differences in genotype distribution between PCOS and controls for rs12086634 or rs6688832 (both P = 0.84). Three per cent of cases and 2.4% of controls had genotype combinations (three or more variant alleles at the two sites) considered characteristic of CRD (P = 0.73). There were no departures from expectation in the family-based association studies, and no significant associations between genotypes (individually or in combination) and BMI, WHR or testosterone. The variants in HSD11B1 and H6PD typed, though implicated in causation of CRD, do not influence susceptibility to PCOS. It seems likely that additional variants within these genes are required for the development of CRD.


Counterpoint: better radiation treatment of non-small cell lung cancer using new techniques without elective nodal irradiation.
The treatment of non-small cell lung cancer has continued to evolve with the advent of improved staging technologies, chemotherapeutic agents, and methods of radiation delivery. Treatment of clinically uninvolved, regional lymph nodes historically has been delivered in the attempt to cover unseen disease, reduce regional failure, and improve survival. None of these suppositions has been tested nor are they supported by data. With enhanced staging using modalities like positron emission tomography and esophageal ultrasonography, treatment portals can be designed to encompass known disease with greater accuracy and confidence. Data for early-stage non-small cell lung cancer is now increasing and strongly suggest that eliminating elective nodal irradiation does not result in a high incidence of nodal relapse and does not compromise survival. Three-dimensional conformal radiotherapy incorporates better targeting and beam directions to effect smaller treatment volumes that include only clinically evident disease. It provides treatment techniques that maximize tumor dose and minimize normal tissue toxicity. Using smaller fields that do not incorporate elective nodal regions may allow higher doses, and these may help improve local control and survival in a disease where current results are unacceptable.


Presentation of three cases followed up with a diagnosis of Felty syndrome.
An effective treatment strategy for Felty syndrome (FS) has not been developed so far. In this article, three cases with FS, who responded to different treatment modalities, have been presented. Case 1 was a 52-year-old male patient who initially received methotrexate, and then, he was switched to granulocyte colony-stimulating factor (G-CSF) and cyclosporine treatment when his neutropenia was further deteriorated. The patient needed monthly doses of G-CSF for nearly 6 months, and his steroid dose was increased. Afterwards, his neutropenia improved with cyclosporine, methotrexate, and hydroxychloroquine combination treatment. Case 2 was a 78-year-old female patient who was started on leflunomide, hydroxychloroquine, and 60 mg methylprednisolone. Case 3 was a 69-year-old female patient who was first treated with 32 mg methylprednisolone, G-CSF, and then with cyclosporine. Neutropenia of both patients improved, and their health status normalized at 2 months. Different treatment strategies have been tried for the management of FS; disease-modifying anti-rheumatic drugs have been used successfully alone or in combination with G-CSF. As seen in the last case, it should be kept in mind that patients can present predominantly with symptoms of infection or hematologic disorders.


Molecular structural dynamics probed by ultrafast X-ray absorption spectroscopy.
The ability to visualize molecular structure in the course of a chemical reaction or a biological function has been a dream of scientists for decades. X-ray absorption spectroscopy (XAS) is ideal in this respect because it is chemically selective and can be implemented in any type of medium. Furthermore, using X-ray absorption near-edge structure (XANES) and extended X-ray absorption fine structure (EXAFS) in laser pump/X-ray probe experiments allows the retrieval of not only the local geometric structure of the system under study, but also the underlying electronic structure changes that drive the structural dynamics. We review recent developments in picosecond and femtosecond XAS applied to molecular systems in solution. Examples on ultrafast photoinduced processes such as intramolecular electron transfer, low-to-high spin change, and bond formation are presented.


Long-term sensory denervation by neonatal capsaicin treatment augments sympathetic neurotransmission in rat mesenteric arteries by increasing levels of norepinephrine and selectively enhancing postjunctional actions.
The present study assessed the long-term effects of sensory denervation on sympathetic innervation in rat mesenteric arteries. Mesenteric arterial beds were isolated from adult rats treated as neonates with capsaicin and from vehicle-treated and untreated rats and perfused at a constant flow rate of 5 ml/min. Frequency-dependent constrictions to electrical field stimulation of sympathetic nerves were markedly augmented in capsaicin-treated rats; maximal constriction was approximately 105% and 169% greater than in vehicle-treated and control preparations, respectively. Maximal contractile responses to norepinephrine (NE) and serotonin (5-HT) were increased by approximately 57% and 85%, respectively, compared with vehicle-treated preparations without a change in the pD2 values. Vasoconstrictions to ATP, vasopressin and KCl were unchanged. In contrast, acute denervation of sensory-motor nerves by in vitro capsaicin treatment had no significant effect on vasoconstrictor responses to electrical field stimulation or to NE, ATP, vasopressin and KCl, although the pD2 value for 5-HT was slightly increased. High-performance liquid chromatographic analysis with electrochemical detection showed an approximately 100% increase in mesenteric arterial NE content after long-term capsaicin treatment. Tissue neuropeptide Y, as assessed by enzyme-linked immunosorbent analysis, was unchanged. In conclusion, long-term sensory denervation of rats produces trophic changes in mesenteric arteries as evidenced by augmented sympathetic vasoconstriction mediated both prejunctionally (increase in tissue NE) and postjunctionally (enhanced responses to NE and 5-HT).


The c-Met receptor tyrosine kinase inhibitor MP470 radiosensitizes glioblastoma cells.
Glioblastoma multiforme (GBM) is resistant to current cytotoxic therapies, in part because of enhanced DNA repair. Activation of the receptor tyrosine kinase c-Met has been shown to protect cancer cells from DNA damage. We hypothesized that inhibiting c-Met would decrease this protection and thus sensitize resistant tumor cells to the effects of radiation therapy. Eight human GBM cell lines were screened for radiosensitivity to the small-molecule c-Met inhibitor MP470 with colony-count assays. Double-strand (ds) DNA breaks was quantified by using antibodies to gamma H2AX. Western blotting demonstrate expression of RAD51, glycogen synthase kinase (GSK)-3beta, and other proteins. A murine xenograft tumor flank model was used for in vivo radiosensitization studies. MP470 reduced c-Met phosphorylation and enhanced radiation-induced cell kill by 0.4 logs in SF767 cells. Cells pretreated with MP470 had more ds DNA damage than cells treated with radiation alone. Mechanistically, MP470 was shown to inhibit dsDNA break repair and increase apoptosis. MP470 influences various survival and DNA repair related proteins such as pAKT, RAD51 and GSK3beta. In vivo, the addition of MP470 to radiation resulted in a tumor-growth-delay enhancement ratio of 2.9 over radiation alone and extended survival time. GBM is a disease site where radiation is often used to address both macroscopic and microscopic disease. Despite attempts at dose escalation outcomes remain poor. MP470, a potent small-molecule tyrosine kinase inhibitor of c-Met, radiosensitized several GBM cell lines both in vitro and in vivo, and may help to improve outcomes for patients with GBM.


Fast hinge detection algorithms for flexible protein structures.
Analysis of conformational changes is one of the keys to the understanding of protein functions and interactions. For the analysis, we often compare two protein structures, taking flexible regions like hinge regions into consideration. The Root Mean Square Deviation (RMSD) is the most popular measure for comparing two protein structures, but it is only for rigid structures without hinge regions. In this paper, we propose a new measure called RMSD considering hinges (RMSDh) and its variant RMSDh(k) for comparing two flexible proteins with hinge regions. We also propose novel efficient algorithms for computing them, which can detect the hinge positions at the same time. The RMSDh is suitable for cases where there is one small hinge region in each of the two target structures. The new algorithm for computing the RMSDh runs in linear time, which is the same as the time complexity for computing the RMSD and is faster than any of previous algorithms for hinge detection. The RMSDh(k) is designed for comparing structures with more than one hinge region. The RMSDh(k) measure considers at most k small hinge region, i.e., the RMSDh(k) value should be small if the two structures are similar except for at most k hinge regions. To compute the value, we propose an O(kn2)-time and O(n)-space algorithm based on a new dynamic programming technique. With the same computational time and space, we can enumerate the predicted hinge positions. We also test our algorithms against actual flexible protein structures, and show that the hinge positions can be correctly detected by our algorithms.


Using deep sequencing to characterize the biophysical mechanism of a transcriptional regulatory sequence.
Cells use protein-DNA and protein-protein interactions to regulate transcription. A biophysical understanding of this process has, however, been limited by the lack of methods for quantitatively characterizing the interactions that occur at specific promoters and enhancers in living cells. Here we show how such biophysical information can be revealed by a simple experiment in which a library of partially mutated regulatory sequences are partitioned according to their in vivo transcriptional activities and then sequenced en masse. Computational analysis of the sequence data produced by this experiment can provide precise quantitative information about how the regulatory proteins at a specific arrangement of binding sites work together to regulate transcription. This ability to reliably extract precise information about regulatory biophysics in the face of experimental noise is made possible by a recently identified relationship between likelihood and mutual information. Applying our experimental and computational techniques to the Escherichia coli lac promoter, we demonstrate the ability to identify regulatory protein binding sites de novo, determine the sequence-dependent binding energy of the proteins that bind these sites, and, importantly, measure the in vivo interaction energy between RNA polymerase and a DNA-bound transcription factor. Our approach provides a generally applicable method for characterizing the biophysical basis of transcriptional regulation by a specified regulatory sequence. The principles of our method can also be applied to a wide range of other problems in molecular biology.


Cortical and vestibular stimulation reveal preserved descending motor pathways in individuals with motor-complete spinal cord injury.
To use a combination of electrophysiological techniques to determine the extent of preserved muscle activity below the clinically-defined level of motor-complete spinal cord injury. Transcranial magnetic stimulation and vestibular-evoked myogenic potentials were used to investigate whether there was any preserved muscle activity in trunk, hip and leg muscles of 16 individuals with motor-complete spinal cord injury (C4-T12) and 16 able-bodied matched controls. Most individuals (14/16) with motor-complete spinal cord injury were found to have transcranial magnetic stimulation evoked, and/or voluntary evoked muscle activity in muscles innervated below the clinically classified lesion level. In most cases voluntary muscle activation was accompanied by a present transcranial magnetic stimulation response. Furthermore, motor-evoked potentials to transcranial magnetic stimulation could be observed in muscles that could not be voluntarily activated. Vestibular-evoked myogenic potentials responses were also observed in a small number of subjects, indicating the potential preservation of other descending pathways. These results highlight the importance of using multiple electrophysiological techniques to assist in determining the potential preservation of muscle activity below the clinically-defined level of injury in individuals with a motor-complete spinal cord injury. These techniques may provide clinicians with more accurate information about the state of various motor pathways, and could offer a method to more accurately target rehabilitation.


Community engagement: The key to tackling Antimicrobial Resistance (AMR) across a One Health context?
Antimicrobial resistance (AMR) is a One Health problem underpinned by complex drivers and behaviours. This is particularly so in low - and middle-income countries (LMICs), where social and systemic factors fuel (mis)use and drive AMR. Behavioural change around antimicrobial use could safeguard both existing and future treatments. However, changing behaviour necessitates engaging with people to understand their experiences. This publication describes a knowledge-exchange cluster of six LMIC-based projects who co-designed and answered a series of research questions around the usage of Community Engagement (CE) within AMR. Findings suggest that CE can facilitate AMR behaviour change, specifically in LMICs, because it is a contextualised approach which supports communities to develop locally meaningful solutions. However, current CE interventions focus on human aspects, and demand-side drivers, of AMR. Our cluster suggests that broader attention should be paid to AMR as a One Health issue. The popularity of mixed methods approaches within existing CE for AMR interventions suggests there is interdisciplinary interest in the uptake of CE. Unfortunately, the specificity and context-dependency of CE can make it difficult to evaluate and scale. Nevertheless, we suggest that in synthesising learnings from CE, we can develop a collective understanding of its scope to tackle AMR across contexts. .


Evaluation of a British computer model to simulate gastrointestinal nematodes in sheep on Canadian farms.
With increasing levels of anthelmintic resistance worldwide and a growing demand to produce more organic products, utilisation of control strategies for gastrointestinal nematodes (GIN) that minimize the use of anthelmintics becomes even more important. This study evaluated the farm-level performance of an existing predictive sheep parasite model from the United Kingdom (UK), using Canadian data. The UK model simulates the epidemiology of three major GIN species of interest (Teladorsagia spp., Haemonchus spp. and Trichostrongylus spp.) and provides a prediction about seasonal parasite levels of lambs and ewes. Model inputs were generated by using data from the first 2 years of a 3-year study (2006-2008) which examined the epidemiology of GIN parasitism in Ontario sheep flocks. Required input data included ewe parasite egg output, pasture-related information and management dynamics. Farm visits in 2006 and 2007 provided relevant data that were collected monthly during the grazing season, on six and seven occasions respectively. These data were collected from 10 ewes and 10 lambs on each farm. For 23 Ontario farms with available data, only 11 farms in 2006 and 14 in 2007 had suitable data to run in the model because the Canadian study was not specifically designed with this simulation model in mind. Observed values for faecal egg counts (FEC) were compared to the model FEC outputs and assessed using linear regression analysis. There was adequate fit between observed and simulated data for 8 of the 11 farms modelled using data generated in 2006 (F=7.55-42.66, df=10-11, R(2)=0.43-0.81, p=0.021 to <0.001) and with 8 of the farms modelled using data generated in 2007 (F=5.56-35.82, df=9-11, R(2)=0.36-0.82, p=0.040 to <0.001). We suggest that the poor fit between observed and simulated data for some data sets may be attributable to low-level infection on farms making regression difficult due to insensitivity of the egg count method at low values, or a pattern for immunity in ewes that contradicted the model assumptions. Required model modifications focused on accommodating the differences between UK and Canadian management styles; specifically the practice of bringing lambs indoors for weaning which was sometimes used on Canadian farms.


Vascular endothelial growth factor as a marker of tumor endothelium.
Vascular endothelial growth factor (VEGF) is an angiogenic growth factor that is a primary stimulant of the vascularization of solid tumors. VEGF production is induced by oncogenic gene mutations in the tumor cells and by hypoxic conditions inside the tumor mass. Hypoxia and the locally increased concentration of VEGF lead to an up-regulation of VEGF receptor expression on tumor endothelial cells. Therefore, in the tumor microenvironment, there is an up-regulation of both VEGF and its receptor, leading to a high concentration of occupied receptor on tumor vascular endothelium. The VEGF:receptor complex presents an attractive target for the specific delivery of drugs or other effectors to tumor endothelium. In the present study, several hybridomas that secrete monoclonal antibodies against the VEGF:receptor (Flk-1) complex or against VEGF itself have been raised. Three of the antibodies (3E7, GV39M, and 11B5) bind with high affinity to the VEGF:Flk-1 complex in ELISA and to tumor endothelium in frozen sections of human tumors, rodent tumors, and human tumor xenografts. 3E7 and GV39M localize selectively to tumor endothelium after i.v. injection into mice bearing human tumor xenografts. Additionally, one antibody (2C3) was raised that blocks the interaction between VEGF and KDR/Flk-1. 2C3 inhibits VEGF-mediated growth of endothelial cells in vitro and localizes strongly to connective tissue in tumors after injection into mice bearing human tumor xenografts. These findings suggest that 3E7, GV39M, and 2C3 are candidates for targeting and imaging the vasculature or connective tissue of tumors.


Continuum tuning of nanoparticle interfacial properties by dynamic covalent exchange.
Surface chemical composition is fundamental to determining properties on the nanoscale, making precise control over surface chemistry critical to being able to optimise nanomaterials for virtually any application. Surface-engineering independent of the preparation of the underlying nanomaterial is particularly attractive for efficient, divergent synthetic strategies, and for the potential to create reactive, responsive and smart nanodevices. For monolayer-stabilised nanoparticles, established methods include ligand exchange to replace the ligand shell in its entirety, encapsulation with amphiphilic (macro)molecules, noncovalent interactions with surface-bound biomolecules, or a relatively limited number of covalent bond forming reactions. Yet, each of these approaches has considerable drawbacks. Here we show that dynamic covalent exchange at the periphery of the nanoparticle-stabilizing monolayer allows surface-bound ligand molecular structure to be substantially modified in mild and reversible processes that are independent of the nanoparticle-molecule interface. Simple stoichiometric variation allows the extent of exchange to be controlled, generating a range of kinetically stable mixed-monolayer compositions across an otherwise identical, self-consistent series of nanoparticles. This approach can be used to modulate nanoparticle properties that are defined by the monolayer composition. We demonstrate switching of nanoparticle solvent compatibility between widely differing solvents - spanning hexane to water - and the ability to tune solubility across the entire continuum between these extremes, all from a single nanoparticle starting point. We also demonstrate that fine control over mixed-monolayer composition influences the assembly of discrete, colloidally stable nanoparticle clusters. By carefully assessing monolayer composition in each state, using both in situ and ex situ methods, we are able to correlate the molecular-level details of the nanoparticle-bound monolayer with system-level properties and behaviour. These empirically determined relationships contribute fundamental insights on nanoscale structure-function relationships, which are currently beyond the capabilities of ab initio prediction.


Community-level socioeconomic status and parental smoking in Japan.
Community-level social environment has been considered to be associated with smoking behavior. However, no study has examined the association between community-level environmental factors and parental smoking behavior in families with young children. The aim of the present study was to examine the association between community-level socioeconomic status (SES) and parental smoking behavior. We used data from a cross-sectional study conducted from 2005 to 2006. We randomly selected 44 Japanese municipalities, 39 of which municipalities agreed to participate in this survey. The study subjects were participants in health check-ups for three-year-old children. Smoking status and individual demographic characteristics were obtained using self-administered questionnaires. Community-level variables were obtained from national census data for 2005. The prevalence of employment in tertiary industries and of unemployment was used to measure community-level SES. Multilevel Poisson regression models were used to calculate prevalence ratios (PRs) for smoking. Of 4143 subjects, a total of 3301 parents in 39 municipalities participated in our survey. Among the 2975 participants (71.8%) included in our analysis, 59.0% were smokers. There was no association between the job of the head of the household considered as an indicator of individual-level SES and smoking. By contrast, when we examined the relationship between prevalence of employment in tertiary industries as community-level SES and smoking, parents living in low middle SES municipalities had a significantly higher prevalence ratio for smoking, compared to parents living in the highest SES municipalities. This result suggested that those with lower community-level SES tended to have a higher prevalence of parental smoking regardless of individual-level SES.


Green revolution: impacts, limits, and the path ahead.
A detailed retrospective of the Green Revolution, its achievement and limits in terms of agricultural productivity improvement, and its broader impact at social, environmental, and economic levels is provided. Lessons learned and the strategic insights are reviewed as the world is preparing a "redux" version of the Green Revolution with more integrative environmental and social impact combined with agricultural and economic development. Core policy directions for Green Revolution 2.0 that enhance the spread and sustainable adoption of productivity enhancing technologies are specified.


Actin kinetics shapes cortical network structure and mechanics.
The actin cortex of animal cells is the main determinant of cellular mechanics. The continuous turnover of cortical actin filaments enables cells to quickly respond to stimuli. Recent work has shown that most of the cortical actin is generated by only two actin nucleators, the Arp2/3 complex and the formin Diaph1. However, our understanding of their interplay, their kinetics, and the length distribution of the filaments that they nucleate within living cells is poor. Such knowledge is necessary for a thorough comprehension of cellular processes and cell mechanics from basic polymer physics principles. We determined cortical assembly rates in living cells by using single-molecule fluorescence imaging in combination with stochastic simulations. We find that formin-nucleated filaments are, on average, 10 times longer than Arp2/3-nucleated filaments. Although formin-generated filaments represent less than 10% of all actin filaments, mechanical measurements indicate that they are important determinants of cortical elasticity. Tuning the activity of actin nucleators to alter filament length distribution may thus be a mechanism allowing cells to adjust their macroscopic mechanical properties to their physiological needs.


The role of thrombosis in severe pulmonary hypertension.
Considering the important surface in pulmonary circulation where blood can interact with the endothelium, the maintenance of blood fluidity through the lung, by antithrombotic pathways and products of the endothelium, is essential. This function appears to be ineffective in primary pulmonary hypertension and in severe secondary pulmonary hypertension. Thrombotic lesions are frequently found in pulmonary arteries in these diseases. Thrombin activity appears to be increased in severe pulmonary hypertension. Antithrombotic pathway disorders may account for this abnormality, particularly in chronic thromboembolic pulmonary hypertension and primary pulmonary hypertension. Injured endothelium, a constant feature in severe pulmonary hypertension, either primary or secondary, enhances thrombus formation in pulmonary vessels. This is probably related to thrombomodulin and tissue factor imbalance, impairment of prostacyclin and nitric oxide release, as well as inefficiency of fibrinolysis. Moreover, platelets appear to be activated in the pulmonary circulation of these patients. They release several mediators acting on vascular tone and as mitogenic agents, and may also contribute to thrombin and clot generation. Long-term oral anticoagulant and continuous infusion of prostacyclin, treatments which impede thrombosis, are known to improve the survival rate in patients with primary pulmonary hypertension. These are the strongest arguments, so far, in favour of the role of thrombosis in severe pulmonary hypertension. However, we do not know whether these abnormalities result from a previous vascular injury or represent the primary disturbance.


Predation on stink bugs (Hemiptera: Pentatomidae) in cotton and soybean agroecosystems.
Stink bugs (Hemiptera: Pentatomidae) are significant pests of cotton and soybeans in the southeastern United States with annual control costs exceeding $14 million in these crops. Three of the most prominent stink bug pests are the southern green (Nezara viridula), brown (Euschistus servus) and green (Chinavia hilaris) stink bugs. To determine trophic linkages between generalist arthropod predators and these pests, species-specific 16S molecular markers were designed and used to detect the presence of prey DNA in predator gut-contents. Over 2700 predators were collected over two growing seasons in cotton and soybean in southern Georgia in 2011 and 2012 and screened for stink bug DNA. Trophic linkages were analyzed relative to prey availability, crop type and field location. The frequency of stink bug DNA in predator guts was negligible on E. servus (0.23%) and C. hilaris (0.09%). Overall gut content detection of N. viridula was 3.3% and Geocoris sp. (Hemiptera: Geocoridae), Orius sp. (Hemiptera: Anthocoridae) and Notoxus monodon (Coleoptera: Anthicidae) were the primary predators. This contrasts with previous studies that reported a much more diverse suite of predators consuming stink bugs with much higher frequency of gut-content positives. The discrepancy between studies highlights the need for replicating studies in space and time, especially if the goal is to implement effective and durable conservation biological control in integrated pest management.


[Physiological (programmed) cell death in hemopoiesis].
According to current concepts, pluripotent cells proliferate and differentiate into "committed" precursors. These committed precursors divide, mature, and give rise to red cells, granulocytes, monocytes, and platelets of the blood. The life span of mature circulating cells being short, and their populations in the blood very stable, a constant and strict regulation of hematopoiesis is needed. The regulation of hematopoiesis is believed to be mediated through precursor cell interaction with specific molecules (glycoproteins) in their microenvironment. Soluble forms of these molecules are termed "hematopoietic growth factors" and include erythropoietin, colony-stimulating factors, interleukins, and stem cell factors. Hematopoietic growth factors not only stimulate the proliferation of precursor cells, but activate the differentiation program and maintain the viability of these cells as well. The normal fate of precursor cells devoid of these factors is programmed suicide. The morphology of such cell death is usually that of apoptosis, rather than of necrosis. The concept of apoptosis was proposed 22 years ago. Apoptosis is a widespread and morphologically distinct process of cell death. The significance of apoptosis stems from its active nature and its ability of controlling biological systems. The present review of published and authors' own data describes apoptosis morphology and presents evidence of the participation of cell reactions of this type in hematopoiesis regulation. The major point in the review is the balance of three normal processes: proliferation, differentiation, and apoptosis, which maintains the homeostasis of hematopoiesis, similarly as of any other cell system; it is well illustrated by recent findings in experimental and clinical hematology.


Meta-analysis and candidate gene mining of low-phosphorus tolerance in maize.
Plants with tolerance to low-phosphorus (P) can grow better under low-P conditions, and understanding of genetic mechanisms of low-P tolerance can not only facilitate identifying relevant genes but also help to develop low-P tolerant cultivars. QTL meta-analysis was conducted after a comprehensive review of the reports on QTL mapping for low-P tolerance-related traits in maize. Meta-analysis produced 23 consensus QTL (cQTL), 17 of which located in similar chromosome regions to those previously reported to influence root traits. Meanwhile, candidate gene mining yielded 215 genes, 22 of which located in the cQTL regions. These 22 genes are homologous to 14 functionally characterized genes that were found to participate in plant low-P tolerance, including genes encoding miR399s, Pi transporters and purple acid phosphatases. Four cQTL loci (cQTL2-1, cQTL5-3, cQTL6-2, and cQTL10-2) may play important roles for low-P tolerance because each contains more original QTL and has better consistency across previous reports.


Medical Physics Residency Consortium: collaborative endeavors to meet the ABR 2014 certification requirements.
In 2009, Mary Bird Perkins Cancer Center (MBPCC) established a Radiation Oncology Physics Residency Program to provide opportunities for medical physics residency training to MS and PhD graduates of the CAMPEP-accredited Louisiana State University (LSU)-MBPCC Medical Physics Graduate Program. The LSU-MBPCC Program graduates approximately six students yearly, which equates to a need for up to twelve residency positions in a two-year program. To address this need for residency positions, MBPCC has expanded its Program by developing a Consortium consisting of partnerships with medical physics groups located at other nearby clinical institutions. The consortium model offers the residents exposure to a broader range of procedures, technology, and faculty than available at the individual institutions. The Consortium institutions have shown a great deal of support from their medical physics groups and administrations in developing these partnerships. Details of these partnerships are specified within affiliation agreements between MBPCC and each participating institution. All partner sites began resident training in 2011. The Consortium is a network of for-profit, nonprofit, academic, community, and private entities. We feel that these types of collaborative endeavors will be required nationally to reach the number of residency positions needed to meet the 2014 ABR certification requirements and to maintain graduate medical physics training programs.


Etiology and pathogenesis of the Marfan syndrome: current understanding.
Much has changed regarding Marfan syndrome (MFS) over the past few decades. Once described solely as a heritable disorder of connective tissue, MFS is now one of a number of conditions recognized to be a disorder of abnormal signalling in the TGF-β pathway. The cardinal features of MFS, once encompassed by the ocular, skeletal and cardiovascular systems, are now known to encompass many more organ systems, especially as people with MFS grow older. They are growing older by several decades compared to the 1970's because of profound improvements in diagnosis and management of the cardiovascular features, especially dilatation of the aortic root. This dilatation can be detected first in infancy and followed up by echocardiography. Progressive enlargement increases the risk of type A dissection and aortic regurgitation, the major causes of early mortality, in untreated patients today. Medical therapy with β-adrenergic blockade, first shown to be effective in the 1980's, can retard this dilatation. In the past decade, angiotensin receptor blockade, which reduces aberrant signalling through one of the TGF-β pathways, also can be effective. However, when dilatation of the root becomes such that the risk of dissection increases to an unacceptable degree, surgical therapy becomes necessary. In the mid-1970's, the composite graft, introduced by Hugh Bentall, markedly reduced mortality. In the past decade, a valve-spring aortic root replacement, advanced by Tirone David, has become widely adopted. Mid-term results are quite encouraging. Other cardiovascular involvement, such as mitral valve prolapse, type B dissection, and dilatation and dissection of aortic branches, also require close monitoring. Currently, life-expectancy in people with MFS who are diagnosed early and treated prophylactically is approaching that of the general population.


Modulation of Wnt signaling by Axin and Axil.
The Wnt signaling pathway is conserved in various species from worms to mammals, and plays important roles in development, cellular proliferation, and differentiation. The molecular mechanisms by which the Wnt signal regulates cellular functions are becoming increasingly well understood. Wnt stabilizes cytoplasmic beta-catenin, which stimulates the expression of genes including c-myc, c-jun, fra-1, and cyclin D1. Axin and its homolog Axil, newly recognized as components of the Wnt signaling pathway, negatively regulate this pathway. Other components of the Wnt signaling pathway, including Dvl, glycogen synthase kinase-3beta (GSK-3beta), beta-catenin, and adenomatous polyposis coli (APC), interact with Axin, and the phosphorylation and stability of beta-catenin are regulated in the Axin complex. Axil has similar functions to Axin. Thus, Axin and Axil act as scaffold proteins in the Wnt signaling pathway, thereby modulating the Wnt-dependent cellular functions.


Late cardiac death in patients undergoing transcatheter aortic valve replacement: incidence and predictors of advanced heart failure and sudden cardiac death.
Little evidence exists of the burden and predictors of cardiac death after transcatheter aortic valve replacement (TAVR). The purpose of this study was to assess the incidence and predictors of cardiac death from advanced heart failure (HF) and sudden cardiac death (SCD) in a large patient cohort undergoing TAVR. The study included a total of 3,726 patients who underwent TAVR using balloon (57%) or self-expandable (43%) valves. Causes of death were defined according to the Valve Academic Research Consortium-2. At a mean follow-up of 22 ± 18 months, 155 patients had died due to advanced HF (15.2% of total deaths, 46.1% of deaths from cardiac causes) and 57 had died due to SCD (5.6% of deaths, 16.9% of cardiac deaths). Baseline comorbidities (chronic obstructive pulmonary disease, atrial fibrillation, left ventricular ejection fraction ≤40%, lower mean transaortic gradient, pulmonary artery systolic pressure >60 mm Hg; p < 0.05 for all) and 2 procedural factors (transapical approach, hazard ratio [HR]: 2.38, 95% confidence interval [CI]: 1.60 to 3.54; p < 0.001; presence of moderate or severe aortic regurgitation after TAVR, HR: 2.79, 95% CI: 1.82 to 4.27; p < 0.001) independently predicted death from advanced HF. Left ventricular ejection fraction ≤40% (HR: 1.93, 95% CI: 1.05 to 3.55; p = 0.033) and new-onset persistent left bundle-branch block following TAVR (HR: 2.26, 95% CI: 1.23 to 4.14; p = 0.009) were independently associated with an increased risk of SCD. Patients with new-onset persistent left bundle-branch block and a QRS duration >160 ms had a greater SCD risk (HR: 4.78, 95% CI: 1.56 to 14.63; p = 0.006). Advanced HF and SCD accounted for two-thirds of cardiac deaths in patients after TAVR. Potentially modifiable or treatable factors leading to increased risk of mortality for HF and SCD were identified. Future studies should determine whether targeting these factors decreases the risk of cardiac death.


Public health impact and cost-effectiveness of the RTS,S/AS01 malaria vaccine: a systematic comparison of predictions from four mathematical models.
The phase 3 trial of the RTS,S/AS01 malaria vaccine candidate showed modest efficacy of the vaccine against Plasmodium falciparum malaria, but was not powered to assess mortality endpoints. Impact projections and cost-effectiveness estimates for longer timeframes than the trial follow-up and across a range of settings are needed to inform policy recommendations. We aimed to assess the public health impact and cost-effectiveness of routine use of the RTS,S/AS01 vaccine in African settings. We compared four malaria transmission models and their predictions to assess vaccine cost-effectiveness and impact. We used trial data for follow-up of 32 months or longer to parameterise vaccine protection in the group aged 5-17 months. Estimates of cases, deaths, and disability-adjusted life-years (DALYs) averted were calculated over a 15 year time horizon for a range of levels of Plasmodium falciparum parasite prevalence in 2-10 year olds (PfPR2-10; range 3-65%). We considered two vaccine schedules: three doses at ages 6, 7·5, and 9 months (three-dose schedule, 90% coverage) and including a fourth dose at age 27 months (four-dose schedule, 72% coverage). We estimated cost-effectiveness in the presence of existing malaria interventions for vaccine prices of US$2-10 per dose. In regions with a PfPR2-10 of 10-65%, RTS,S/AS01 is predicted to avert a median of 93,940 (range 20,490-126,540) clinical cases and 394 (127-708) deaths for the three-dose schedule, or 116,480 (31,450-160,410) clinical cases and 484 (189-859) deaths for the four-dose schedule, per 100,000 fully vaccinated children. A positive impact is also predicted at a PfPR2-10 of 5-10%, but there is little impact at a prevalence of lower than 3%. At $5 per dose and a PfPR2-10 of 10-65%, we estimated a median incremental cost-effectiveness ratio compared with current interventions of $30 (range 18-211) per clinical case averted and $80 (44-279) per DALY averted for the three-dose schedule, and of $25 (16-222) and $87 (48-244), respectively, for the four-dose schedule. Higher ICERs were estimated at low PfPR2-10 levels. We predict a significant public health impact and high cost-effectiveness of the RTS,S/AS01 vaccine across a wide range of settings. Decisions about implementation will need to consider levels of malaria burden, the cost-effectiveness and coverage of other malaria interventions, health priorities, financing, and the capacity of the health system to deliver the vaccine. PATH Malaria Vaccine Initiative; Bill & Melinda Gates Foundation; Global Good Fund; Medical Research Council; UK Department for International Development; GAVI, the Vaccine Alliance; WHO.


The ascending pathophysiology of cholestatic liver disease.
In this review we develop the argument that cholestatic liver diseases, particularly primary biliary cholangitis and primary sclerosing cholangitis (PSC), evolve over time with anatomically an ascending course of the disease process. The first and early lesions are in "downstream" bile ducts. This eventually leads to cholestasis, and this causes bile salt (BS)-mediated toxic injury of the "upstream" liver parenchyma. BS are toxic in high concentration. These concentrations are present in the canalicular network, bile ducts, and gallbladder. Leakage of bile from this network and ducts could be an important driver of toxicity. The liver has a great capacity to adapt to cholestasis, and this may contribute to a variable symptom-poor interval that is often observed. Current trials with drugs that target BS toxicity are effective in only about 50%-60% of primary biliary cholangitis patients, with no effective therapy in PSC. This motivated us to develop and propose a new view on the pathophysiology of primary biliary cholangitis and PSC in the hope that these new drugs can be used more effectively. These views may lead to better stratification of these diseases and to recommendations on a more "tailored" use of the new therapeutic agents that are currently tested in clinical trials. Apical sodium-dependent BS transporter inhibitors that reduce intestinal BS absorption lower the BS load and are best used in cholestatic patients. The effectiveness of BS synthesis-suppressing drugs, such as farnesoid X receptor agonists, is greatest when optimal adaptation is not yet established. By the time cytochrome P450 7A1 expression is reduced these drugs may be less effective. Anti-inflammatory agents are probably most effective in early disease, while drugs that antagonize BS toxicity, such as ursodeoxycholic acid and nor-ursodeoxycholic acid, may be effective at all disease stages. Endoscopic stenting in PSC should be reserved for situations of intercurrent cholestasis and cholangitis, not for cholestasis in end-stage disease. These are arguments to consider a step-wise pathophysiology for these diseases, with therapy adjusted to disease stage. An obstacle in such an approach is that disease stage-defining biomarkers are still lacking. This review is meant to serve as a call to prioritize the development of biomarkers that help to obtain a better stratification of these diseases. (Hepatology 2017;65:722-738).