Effect of eggshell temperature and oxygen concentration during incubation on the developmental and physiological status of broiler hatchlings in the perinatal period.
This study evaluated the influence of incubation conditions on the developmental and physiological status of birds in the perinatal period, which spans the end of incubation until the early posthatch period. Embryos were incubated at a normal (37.8°C) or high (38.9°C) eggshell temperature (EST) and a low (17%), normal (21%), or high (25%) O(2) concentration from d 7 until 19 of incubation. After d 19 of incubation, EST was maintained, but O(2) concentrations were 21% for all embryos. Body and organ weights, and hepatic glycogen levels were measured at d 18 of incubation and at 12 and 48 h after emergence from the eggshell. In addition, blood metabolites were measured at 12 and 48 h after emergence from the eggshell. Embryos incubated at a high EST and low O(2) concentration had the highest mortality in the last week of incubation, which may be related to their low yolk-free body mass (YFBM) or a reduced nutrient availability for hatching (i.e., hepatic glycogen). High EST, compared with normal EST, decreased YFBM. This may be due to the shorter incubation duration of 8 h, the lower weight of supply organs (i.e., heart and lung), or a lack of glucose precursors. Because of this lack of glucose precursors, embryos incubated at high EST may have used proteins for energy production instead of for body development at the end of incubation. The YFBM at d 18 of incubation increased with an increase in O(2) concentration. However, differences between the normal and high O(2) concentration disappeared at 12 and 48 h after emergence, possibly because the high O(2) concentration had difficulties adapting to lower O(2) concentrations in the perinatal period. Blood metabolites and hepatic glycogen were comparable among O(2) concentrations, indicating that the physiological status at hatch may be related to the environment that the embryo experienced during the hatching process. In conclusion, EST and O(2) concentration differentially influence the developmental and physiological status of broilers during the perinatal period.


Chiral Generation of Hot Carriers for Polarization-Sensitive Plasmonic Photocatalysis.
Mastering the manipulation of chirality at the nanoscale has long been a priority for chemists, physicists, and materials scientists, given its importance in the biochemical processes of the natural world and in the development of novel technologies. In this vein, the formation of novel metamaterials and sensing platforms resulting from the synergic combination of chirality and plasmonics has opened new avenues in nano-optics. Recently, the implementation of chiral plasmonic nanostructures in photocatalysis has been proposed theoretically as a means to drive polarization-dependent photochemistry. In the present work, we demonstrate that the use of inorganic nanometric chiral templates for the controlled assembly of Au and TiO2 nanoparticles leads to the formation of plasmon-based photocatalysts with polarization-dependent reactivity. The formation of plasmonic assemblies with chiroptical activities induces the asymmetric formation of hot electrons and holes generated via electromagnetic excitation, opening the door to novel photocatalytic and optoelectronic features. More precisely, we demonstrate that the reaction yield can be improved when the helicity of the circularly polarized light used to activate the plasmonic component matches the handedness of the chiral substrate. Our approach may enable new applications in the fields of chirality and photocatalysis, particularly toward plasmon-induced chiral photochemistry.


Preparation and characterization of stable phospholipid-silica nanostructures loaded with quantum dots.
The structural dependence of silica-liposome hybrids on silanization conditions was investigated. Silica coatings protect liposomes against aggregation, degradation, and leakage, which are important for their application in bioimaging. Liposomes loaded with quantum dots were synthesized and attempts to obtain uniformly sized, silica-coated nanocapsules were made.


SnRK1 from Arabidopsis thaliana is an atypical AMPK.
SNF1-related protein kinase 1 (SnRK1) is the plant orthologue of the evolutionarily-conserved SNF1/AMPK/SnRK1 protein kinase family that contributes to cellular energy homeostasis. Functional as heterotrimers, family members comprise a catalytic α subunit and non-catalytic β and γ subunits; multiple isoforms of each subunit type exist, giving rise to various isoenzymes. The Arabidopsis thaliana genome contains homologues of each subunit type, and, in addition, two atypical subunits, β(3) and βγ, with unique domain architecture, that are found only amongst plants, suggesting atypical heterotrimers. The AtSnRK1 subunit structure was determined using recombinant protein expression and endogenous co-immunoprecipitation, and six unique isoenzyme combinations were identified. Each heterotrimeric isoenzyme comprises a catalytic α subunit together with the unique βγ subunit and one of three non-catalytic β subunits: β(1), β(2) or the plant-specific β(3) isoform. Thus, the AtSnRK1 heterotrimers contain the atypical βγ subunit rather than a conventional γ subunit. Mammalian AMPK heterotrimers are phosphorylated on the T-loop (pThr175/176) within both catalytic a subunits. However, AtSnRK1 is insensitive to AMP and ADP, and is resistant to T-loop dephosphorylation by protein phosphatases, a process that inactivates other SNF1/AMPK family members. In addition, we show that SnRK1 is inhibited by a heat-labile, >30 kDa, soluble proteinaceous factor that is present in the lysate of young rosette leaves. Finally, none of the three SnRK1 carbohydrate-binding modules, located in the β(1), β(2) and βγ subunits, associate with various carbohydrates, including starch, the plant analogue of glycogen to which AMPK binds in vitro. These data clearly demonstrate that AtSnRK1 is an atypical member of the SNF1/AMPK/SnRK1 family.


The Future of Cryopreservation in Assisted Reproductive Technologies.
Societal changes and the increasing desire and opportunity to preserve fertility have increased the demand for effective assisted reproductive technologies (ART) and have increased the range of scenarios in which ART is now used. In recent years, the "freeze-all" strategy of cryopreserving all oocytes or good quality embryos produced in an IVF cycle to transfer later-at a time that is more appropriate for reasons of medical need, efficacy, or desirability-has emerged as an accepted and valuable alternative to fresh embryo transfer. Indeed, improvements in cryopreservation techniques (vitrification) and the development of more efficient ovarian stimulation protocols have facilitated a dramatic increase in the practice of elective frozen embryo transfer (eFET). Alongside these advances, debate continues about whether eFET should be a standard treatment option available to the whole IVF population or if it is important to identify patient subgroups who are most likely to benefit from such an approach. Achieving successful outcomes in ART, whether by fresh or frozen embryo transfer, is influenced by a wide range of factors. As well as the efficiency of IVF and embryo transfer protocols and techniques, factors affecting implantation include maternal aging, sperm quality, the vaginal and endometrial microbiome, and peri-implantation levels of serum progesterone. The safety of eFET, both during ART cycles and on longer-term obstetric and neonatal outcomes, is also an important consideration. In this review, we explore the benefits and risks of freeze-all strategies in different scenarios. We review available evidence on the outcomes achieved with elective cryopreservation strategies and practices and how these compare with more traditional IVF cycles with fresh embryo transfers, both in the general IVF population and in subgroups of special interest. In addition, we consider how to optimize and individualize "freeze-all" procedures to achieve successful reproductive outcomes.


SRF-dependent gene expression in isolated cardiomyocytes: regulation of genes involved in cardiac hypertrophy.
Serum response factor (SRF) is a transcription factor required for the regulation of genes important for cardiac structure and function. Notably, the "fetal gene expression profile" that is characteristic of cardiac hypertrophy consists of genes known to be regulated by SRF. Transgenic animal studies suggest that cardiac-specific overexpression of SRF induces this pattern of hypertrophic genes and subsequently causes the progression of pathologic adaptations. Furthermore, studies examining cardiac tissues from patients with severe heart failure indicate significant alterations in SRF expression that correspond with alterations in expression of SRF-dependent genes. Based on these observations, it has been postulated that SRF may be critical for stimulating pathologic gene expression at the onset of hypertrophic adaptation. To address the role of SRF in cardiac hypertrophy we investigated whether SRF is necessary and sufficient for the expression of genes associated with the hypertrophic response. We used isolated cardiomyocytes from both neonatal rats, and transgenic mice containing floxed SRF alleles, to examine cardiac gene expression in response to overexpression and absence of SRF. Using this approach, we demonstrate that SRF is required for the induction of atrial naturetic factor (ANF), c-fos, NCX1, BNP, alpha-actins, alpha-myosin heavy chain, and beta-myosin heavy chain genes. However, overexpression of exogenous SRF in isolated cardiomyocytes is only sufficient to induce NCX1 and alpha-myosin heavy chain. These results indicate that SRF is critical for the regulation and induction of genes associated with the progression of pathologic cardiac hypertrophy, however, the pattern of genes induced by overexpression of SRF in isolated cardiomyocytes is different from those genes expressed in hypertrophic transgenic hearts. This suggests that SRF-dependent gene expression is modulated in a complex manner by in vivo physiologic systems prior to and during heart failure as the organism adapts to cardiac stress.


A hybrid discrete-continuum mathematical model of pattern prediction in the developing retinal vasculature.
Pathological angiogenesis has been extensively explored by the mathematical modelling community over the past few decades, specifically in the contexts of tumour-induced vascularisation and wound healing. However, there have been relatively few attempts to model angiogenesis associated with normal development, despite the availability of animal models with experimentally accessible and highly ordered vascular topologies: for example, growth and development of the vascular plexus layers in the murine retina. The current study aims to address this issue through the development of a hybrid discrete-continuum mathematical model of the developing retinal vasculature in neonatal mice that is closely coupled with an ongoing experimental programme. The model of the functional vasculature is informed by a range of morphological and molecular data obtained over a period of several days, from 6 days prior to birth to approximately 8 days after birth. The spatio-temporal formation of the superficial retinal vascular plexus (RVP) in wild-type mice occurs in a well-defined sequence. Prior to birth, astrocytes migrate from the optic nerve over the surface of the inner retina in response to a chemotactic gradient of PDGF-A, formed at an earlier stage by migrating retinal ganglion cells (RGCs). Astrocytes express a variety of chemotactic and haptotactic proteins, including VEGF and fibronectin (respectively), which subsequently induce endothelial cell sprouting and modulate growth of the RVP. The developing RVP is not an inert structure; however, the vascular bed adapts and remodels in response to a wide variety of metabolic and biomolecular stimuli. The main focus of this investigation is to understand how these interacting cellular, molecular, and metabolic cues regulate RVP growth and formation. In an earlier one-dimensional continuum model of astrocyte and endothelial migration, we showed that the measured frontal velocities of the two cell types could be accurately reproduced by means of a system of five coupled partial differential equations (Aubert et al. in Bull. Math. Biol. 73:2430-2451, 2011). However, this approach was unable to generate spatial information and structural detail for the entire retinal surface. Building upon this earlier work, a more realistic two-dimensional hybrid PDE-discrete model is derived here that tracks the migration of individual astrocytes and endothelial tip cells towards the outer retinal boundary. Blood perfusion is included throughout plexus development and the emergent retinal architectures adapt and remodel in response to various biological factors. The resulting in silico RVP structures are compared with whole-mounted retinal vasculatures at various stages of development, and the agreement is found to be excellent. Having successfully benchmarked the model against wild-type data, the effect of transgenic over-expression of various genes is predicted, based on the ocular-specific expression of VEGF-A during murine development. These results can be used to help inform future experimental investigations of signalling pathways in ocular conditions characterised by aberrant angiogenesis.


Regulation of the autoimmune plaque-forming cell response to single-strand DNA (sDNA) in vitro.
We have shown that young autoimmune and normal strain mice possess autoantigen-sensitive cells potentially capable of producing anti-sDNA autoantibody in the absence of normal regulatory mechanisms in vitro. In certain strains such as B/W mice, these regulatory mechanisms presumably break down with increasing age, and autoimmunity develops. These regulatory mechanisms might consist of sDNA, T cells, or some combination of these since both of these agents suppressed the anti-sDNA PFC response in vitro. The sDNA may have inhibited PFC development by a receptor blockade mechanism since i) spleen cells pulsed with sDNA for short periods and then washed were suppressed after 5 days of culture; ii) treatment of these blocked cells with trypsin and DNase I restored the anti-sDNA response; iii) the PFC remaining in partially blocked cultures were of lower avidity than PFC in unblocked cultures; and iv) the target of sDNA may be a B cell. Thymocytes and splenic T cells suppressed the anti-sDNA response but not the anti-SRBC response in vitro in a dose-dependent manner. The suppressive capacity of thymus cells did not decline with age in B/W mice. In addition, thymus cells activated by competing foreign antigens could also suppress the anti-sDNA response. The relationship between these modes of regulating autoreactivity remains to be investigated.


What Is the Role of Steroids for Septic Shock in 2021?
Corticosteroids have been used for decades in the adjunctive treatment of severe infections in intensive care. The most frequent scenario in intensive care is in septic shock, where low doses of glucocorticoids appear to restore vascular responsiveness to norepinephrine. There is a strong body of evidence suggesting that hydrocortisone reduces time on vasopressor, and may modulate the immune response. In this review, we explore the current evidence supporting the use of corticosteroids in septic shock, its benefits, and potential harms. In addition to landmark clinical trials, we will also describe new frontiers for the use of corticosteroids in septic shock which should be explored in future studies.


2D IR spectroscopy of high-pressure phases of ice.
We present experimental and simulated 2D IR spectra of some high-pressure forms of isotope-pure D2O ice and compare the results to those of ice Ih published previously [F. Perakis and P. Hamm, Phys. Chem. Chem. Phys. 14, 6250 (2012); L. Shi et al., ibid. 18, 3772 (2016)]. Ice II, ice V, and ice XIII have been chosen for this study, since this selection covers many aspects of the polymorphism of ice. That is, ice II is a hydrogen-ordered phase of ice, in contrast to ice Ih, while ice V and ice XIII are a hydrogen-disordered/ordered couple that shares essentially the same oxygen structure and hydrogen-bonded network. For the transmission 2D IR spectroscopy, a novel method had to be developed for the preparation of ultrathin films (1-2 μm) of high-pressure ices with good optical quality. We also simulated 2D IR spectra based on molecular dynamics simulations connected to a vibrational exciton picture. These simulations agree with the experimental results in a semi-quantitative manner for ice II, while the same approach failed for ice V and ice XIII. From the perspective of 2D IR spectroscopy, ice II appears to be more inhomogeneously broadened than ice Ih, despite its hydrogen-order, which we attribute to the fact that ice II is structurally more complex with four distinguishable hydrogen bonds that mix due to exciton coupling. Ice V and ice XIII, on the other hand, behave as expected with the hydrogen-disordered case (ice V) being more inhomogenously broadened. Furthermore, in all hydrogen-ordered forms (ice II and ice XIII), cross peaks could be identified in the anisotropic 2D IR spectrum, whose signs reveal the relative direction of the corresponding excitonic states.


Nanoribbons with Nonalternant Topology from Fusion of Polyazulene: Carbon Allotropes beyond Graphene.
Various two-dimensional (2D) carbon allotropes with nonalternant topologies, such as pentaheptites and phagraphene, have been proposed. Predictions indicate that these metastable carbon polymorphs, which contain odd-numbered rings, possess unusual (opto)electronic properties. However, none of these materials has been achieved experimentally due to synthetic challenges. In this work, by using on-surface synthesis, nanoribbons of the nonalternant graphene allotropes, phagraphene and tetra-penta-hepta(TPH)-graphene, have been obtained by dehydrogenative C-C coupling of 2,6-polyazulene chains. These chains were formed in a preceding reaction step via on-surface Ullmann coupling of 2,6-dibromoazulene. Low-temperature scanning probe microscopies with CO-functionalized tips and density functional theory calculations have been used to elucidate their structural properties. The proposed synthesis of nonalternant carbon nanoribbons from the fusion of synthetic line-defects may pave the way for large-area preparation of novel 2D carbon allotropes.


Prediction of forest aboveground net primary production from high-resolution vertical leaf-area profiles.
Temperature and precipitation explain about half the variation in aboveground net primary production (ANPP) among tropical forest sites, but determinants of remaining variation are poorly understood. Here, we test the hypothesis that the amount of leaf area, and its vertical arrangement, predicts ANPP when other variables are held constant. Using measurements from airborne lidar in a lowland Neotropical rain forest, we quantify vertical leaf-area profiles and develop models of ANPP driven by leaf area and other measurements of forest structure. Vertical leaf-area profiles predict 38% of the variation among plots. This number is 4.5 times greater than models using total leaf area (disregarding vertical arrangement) and 2.1 times greater than models using canopy height alone. Furthermore, ANPP predictions from vertical leaf-area profiles were less biased than alternate metrics. Variation in ANPP not attributable to temperature or precipitation can be predicted by the vertical distribution of leaf area in this system.


Propinox in biliary colic: a multicenter, randomized, prospective and parallel double-blind study of three doses of propinox versus placebo in acute biliary colic pain.
The aim of this study was to assess the efficacy and tolerance of propinox administered i.v., and establish a dose-response relation according to three dose levels (10, 20 and 30 mg), vs. placebo in patients with moderate to severe acute biliary pain. Three hundred and fifty patients were included: 85 received placebo treatment, 81 were treated with propinox 10 mg, 91 with propinox 20 mg and 93 received propinox 30 mg. Spontaneous pain intensity was assessed according to a visual analog and a verbal scale before treatment and 20, 60 and 120 min after. All treatments induced significant and progressive pain reduction at all controls, but patients treated with 20 and 30 mg of propinox showed significantly lower pain intensity after 120 min compared to the placebo group. The last control revealed that 28% of patients receiving placebo had no pain while 60% of patients treated with propinox 30 mg reported absence of pain with a statistically significant difference (p < 0.001). All treatments were very well tolerated and there were no dropouts due to adverse events. Mouth dryness was the adverse effect occurring with a significantly higher frequency than that observed with placebo although it was only seen in patients treated with 20 mg and 30 mg active doses. The results of this study showed that propinox was an effective drug in the treatment of moderate to severe colic pain of biliary origin. Concerning efficacy and side effects, a clear dose-response relation was observed; the 20 mg and 30 mg doses being significantly superior to placebo.


Computerized transaxial x-ray tomography of the human body.
The ACTA-Scanner has virtually unlimited potential in the evalution of any part of the body. The usefulness of the technique has already been shown in the appraisal of pathologies of the brain and cerebrospinal fluid cavities. The orbits and the eyeballs, the facial sinuses, and skull base lesions have also been elucidated. Tumors of the larynx, pharynx, thyroid, and parathyroid; lymphomas; and pathology of the spine and spinal cord are well within the reach of this new diagnostic methodology. Lung pathologies, such as emphysema, pneumonias, neoplasms, infarctions, pleural effusions and granulomatous diseases, and mediastinal pathology represent a challenging complex of lesions to be appraised by ACTA-scanning. For the heart, there is great potential for observing cardiac chamber size, hypertrophy of ventricular or atrial walls, and ventricular or aortic aneurysms, and possibly for recognizing the damaged myocardial tissue immediately after or some time after an infarction. The abdominal pathologies that can be studied are almost uncountable: gastric neoplasms, pancreatic cysts and stones, gallstones, neoplasms of the liver and pancreas, bowel tumors, abdominal aortic aneurysms, renal neoplasms and cysts, atrophy of the kidneys, bladder tumors, uterine tumors, ovarian cysts, and many more. Although bones and joints are adequately demonstrated by conventional x-ray techniques, there is no doubt that as the new technique is developed ACTA-grams will contribute significant information in the transverse plane, as well as in densitometric analyses. The impact of ACTA-scanning will not be limited to the diagnostic area, but will extend, at least indirectly, to general patient management and to some aspects of medical economics as well. Risk-laden, technically complex, and costly diagnostic procedures, sometimes requiring lengthy hospitalization, will in some cases be eliminated. The simple, innocuous, and noninvasive ACTA-scanning can be performed on an outpatient basis. Repeated follow-up examinations should be easily accepted by the patients, considering that this diagnostic test is carried out without discomfort. The entire field of diagnostic radiology is on the verge of revolutionary changes.


Generation of nano- and micro-sized organic pollutant emulsions in simulated road runoff.
A wide range of organic pollutants (OPs) are emitted from the road and traffic environment and transported with road runoff to receiving waters. To provide an understanding of the transport routes of OPs in the environment, an investigation was carried out with the aim to determine whether OPs are transported with nano- and microparticles in the form of emulsions. Tests were performed on simulated road runoff, using laboratory prepared mixtures of ultrapure water and specific polycyclic aromatic hydrocarbons (PAHs), alkylphenols (APs) and their ethoxylates (APEOs), phthalates, diesel oil (aliphatic hydrocarbons), with and without addition of humic acid (HA) and iron (Fe) colloids. The samples were analysed directly after mixing and after a few days of stabilisation for particle size distribution (PSD) and concentrations of particles in the size range 10 nm-100 μm, and zeta potential > ± 500 mV. Further, after long-term storage to achieve stabilisation, selected samples were investigated for the PSD and particle concentrations in the ranges 10 nm-2 μm, to determine whether stable emulsions had formed. The following simulation mixtures, both mixed and stabilised, were identified as potential emulsions: diesel, APs and APEOs, diesel with APs and APEOS, phthalates, and a mixture of all OPs with and without colloids. Measurements with the Zetasizer and Nanosight instruments imply that the majority of particles in the samples were found in the nano-range of 30-660 nm respectively, and a smaller portion of particles < 28% also measured with Coulter Counter were found to be micro-sized. Higher concentrations of the smallest nanoparticles were found in the mixture of all OPs without colloids added, than in the OP mixture with colloids added. The results indicate that the addition of colloids favours the formation of larger micro-sized emulsions that may break down with time into nano-sized particles. In the mixed samples, the number of micro-sized particles decreased, while the number of nanoparticles increased; this process may also occur in road runoff transportation systems during heavy rain events. This is the first study to indicate that emulsions of OPs may be formed in road runoff, and that emulsions may act as carriers of OPs in urban stormwater.


Poisson-process limit laws yield Gumbel max-min and min-max.
"A chain is only as strong as its weakest link" says the proverb. But what about a collection of statistically identical chains: How long till all chains fail? The answer to this question is given by the max-min of a matrix whose (i,j) entry is the failure time of link j of chain i: take the minimum of each row, and then the maximum of the rows' minima. The corresponding min-max is obtained by taking the maximum of each column, and then the minimum of the columns' maxima. The min-max applies to the storage of critical data. Indeed, consider multiple backup copies of a set of critical data items, and consider the (i,j) matrix entry to be the time at which item j on copy i is lost; then, the min-max is the time at which the first critical data item is lost. In this paper we address random matrices whose entries are independent and identically distributed random variables. We establish Poisson-process limit laws for the row's minima and for the columns' maxima. Then, we further establish Gumbel limit laws for the max-min and for the min-max. The limit laws hold whenever the entries' distribution has a density, and yield highly applicable approximation tools and design tools for the max-min and min-max of large random matrices. A brief of the results presented herein is given in: Gumbel central limit theorem for max-min and min-max [Eliazar, Metzler, and Reuveni, Phys. Rev. E 100, 020104 (2019)10.1103/PhysRevE.100.020104].


Synthesis of carbazole-based BODIPY dimers showing red fluorescence in the solid state.
Carbazole-based BODIPY dimers 2a-g were synthesized via direct arylation. They showed red-shifted solid-state fluorescence spectra as compared with the corresponding monomer. In addition, unsymmetrical dimers 2d, 2f, and 2g with two different substituents showed red fluorescence with improved quantum yields in the solid state.


Analyses of black Aspergillus species of peanut and maize for ochratoxins and fumonisins.
The genus Aspergillus section Nigri, or the black aspergilli, represents genetically closely related species that produce the mycotoxins, ochratoxins and the fumonisins. Fumonisin B1 (FB1) is of an added concern because it is also a virulence factor for maize. Our preliminary data indicated that black aspergilli could develop asymptomatic infections with maize and peanuts plants. Symptomless infections are potential problems, because under favorable conditions, there is a potential for accumulation of ochratoxins and the fumonisins in contaminated postharvest crops. In the present report, the ability of black aspergilli from peanuts and maize to produce ochratoxin A and FB1 on maize kernels was assessed. One hundred fifty strains from peanuts and maize were isolated from several southeastern and midwestern states. Aspergillus nigri (A. nigri var. nigri) was the dominant species (87%), while Aspergillus foetidus, Aspergillus japonicus, Aspergillus tubingensis, and Aspergillus carbonarius were infrequently isolated. None of the wild isolates produced detectable amounts of ochratoxins. However, we do report the occurrence of the fumonisins B1, B2, and B3. Of 54 field isolates, 30% (n = 16) produced FB1, 61% (n = 33) produced FB2, and 44% (n = 24) produced FB3. The amounts of fumonisins produced during the test period of 30 days suggest that these strains might be weak to moderate producers of fumonisin on maize. To our knowledge, this is a first report of FB1 and FB3 production by isolates of black aspergilli from an American cereal and legume.


Effect of reresection in extremity soft tissue sarcoma.
To determine whether reresection affects survival in patients with inadequately resected, primary extremity soft tissue sarcoma. This study correlates reresection with local recurrence-free survival, metastasis-free survival, and disease-free survival. Soft tissue sarcomas are rare neoplasms, with an incidence of approximately 6,000 per year in the United States. Because these tumors are rare and benign soft tissue tumors are common, many are initially thought to be benign and are excised without wide margins. Patients who underwent treatment for primary tumors from July 1982 to June 1999 at a single institution were the subject of study. Two groups of patients were analyzed: those who underwent one definitive resection (one operation) and those whose tumors were previously resected and who were then referred for subsequent reresection (two operations). Patients were given adjuvant radiation or chemotherapy according to the standard of care. Of 1,092 patients with primary extremity soft tissue sarcoma underwent resection, 685 underwent definitive radical resection and 407 underwent reresection after undergoing excisional resection elsewhere. Median follow-up was 4.8 years. The 5-year disease-free survival rate of the definitive resection (one operation) group was 70%; that of the reresection (two operations) group was 88%. On multivariate analysis, reresection was adjusted and controlled for age, grade, depth, size, histology, and margins. Reresection remained a significant predictor of improved disease-free survival, even after these adjustments. To determine whether this difference was stage- or referral-biased, the patient population was divided by AJCC stage. In all stages there was a trend toward improved outcome; this was most marked for those with stage III disease (>5 cm, high-grade, and deep). Patients with extremity soft tissue sarcoma who undergo reresection with two "primary" operations have an improved survival compared with those who undergo one operation. The most plausible explanation, referral and selection bias, is questionable given the significance of reresection as a variable after adjusting for stage and other risk factors. This suggests that where indicated and possible, reresection should be liberally applied in patients with primary extremity soft tissue sarcoma.


[Direct CT-arthrography versus direct MR-arthrography in chronic shoulder instability: comparison of modalities after the introduction of multidetector-CT technology].
To evaluate the diagnostic accuracy of direct multidetector CT arthrography (CTA) and direct MR arthrography (MRA) in patients suffering from chronic shoulder instability. Twenty-nine patients suffering from chronic shoulder instability were included into a prospective study. In all cases, the indication for direct CTA and arthroscopy was set by the orthopedic surgeon. Prior to the imaging procedures, 10 to 20 ml of a special combination of contrast media (including saline, Isovist(R) and Magnevist(R) in a relation of 125 : 125 : 1) was injected into the joint under sterile conditions. First, CTA was performed with a multidetector CT, with images reconstructed in the axial, semi-coronal and semi-sagittal planes. Thereafter, MRA was performed. Axial images were obtained using a T1-weighted, fat-saturated spin echo sequence and semi-coronal images using a T1-weighted FLASH-3D GRE sequence. The results of CTA and MTA were compared with results obtained from arthroscopy or arthrotomy. MRA was superior to CTA in the detection of labral lesions. The sensitivity of MRA was 96 % and the specificity 96 %, compared to a sensitivity of 76 % (p < 0.05) and specificity of 92 % for CTA. Both methods showed the same effectiveness concerning the assessment of capsule distension (sensitivity for both techniques: 91 %). MRA seems to be superior to CTA in the diagnostic workup of chronic shoulder instability even when using a multidetector CT technique.


[Clinical features of early alcoholism in adolescents with a slight handicap of exogenous etiology].
On the basis of clinico-psychopathological and experimental-psychological examination of male children and adolescents with general mental retardation of exogenous etiology and the signs of alcoholism of different severity, the authors describe the diagnostic importance of additional symptoms of alcoholism. They also consider correlations between the age at the beginning of systematic alcohol abuse and the clinical picture of the first states of intoxication with the clinico-physiological form of oligophrenia. The data on the psychopathological manifestations of the major syndromes of early alcoholism in the course of disease are presented.


Ultrasonographic diagnosis of suspected hemopneumothorax in trauma patients.
Bleeding and trapped air in the pleural space are called hemothorax and pneumothorax, respectively. In cases where there are delays in diagnosis and treatment, the mortality rates due to hemopneumothorax can be significant. Hemopneumothorax is characterized by decreased lung sounds or chest percussion and subcutaneous emphysema. Diagnosis of pneumothorax and hemothorax can be achieved by portable chest X-ray (CXR), computed tomography (CT) scan, or ultrasonography. Portable CXR and CT-scans have their individual drawbacks. CXR creates a high percentage of false negative results, and a CT-scan is time consuming and less cost-effective; in addition, both modalities expose patients to radiation. Therefore, the introduction of ultrasonography as an easily available and highly accurate diagnostic modality has particular importance. The aim of this study was to evaluate the sensitivity and specificity of ultrasonography in the diagnosis of pneumothorax and hemothorax in comparison with the other two methods, namely portable CXR and CT-scan. Patients (163) with multiple trauma who were suspected of having chest injuries, and who had indications for a chest CT-scan according to ATLS algorithms, were included in the study. All patients underwent portable CXR, CT-scan, and ultrasonography. In total, 163 patients were included in this study; 29 patients had a pneumothorax, 24 patients had a hemothorax, and 23 patients had a hemopneumothorax confirmed. The study revealed that ultrasonography had a sensitivity of 96.15%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 98%, in the diagnosis of pneumothorax. The sensitivity for ultrasonography in the diagnosis of a hemothorax was 82.97%, with a specificity of 98.05%, a positive predictive value of 90%, and a negative predictive value of 92.66%. Portable CXR for pneumothorax detection had a sensitivity of 34.61%, a specificity of 97.95%, a positive predictive value of 90%, and a negative predictive value of 73.84%. In the detection of hemothorax, CXR had a sensitivity of 25.53%, a specificity of 95.14%, a positive predictive value of 70.58%, and a negative predictive value of 73.68%. Ultrasonography sensitivity and specificity for diagnosis of hemopneumothorax was high. The sensitivity of portable CXR was low despite its high specificity for the detection of hemothorax and pneumothorax.


Does Drought Increase the Risk of Insects Developing Behavioral Resistance to Systemic Insecticides?
Increases in severity and frequency of drought periods, average global temperatures, and more erratic fluctuations in rainfall patterns due to climate change are predicted to have a dramatic impact on agricultural production systems. Insect pest populations in agricultural and horticultural systems are also expected to be impacted, both in terms of their spatial and temporal distributions and in their status as pest species. In this opinion-based article, we discuss how indirect effects of drought may adversely affect the performance of systemic insecticides and also lead to increased risk of insect pests developing behavioral insecticide resistance. We hypothesize that more pronounced drought will decrease uptake and increase the magnitude of nonuniform translocation of systemic insecticides within treated crop plants, and that may have two concurrent consequences: 1) reduced pesticide performance, and 2) increased likelihood of insect pests evolving behavioral insecticide resistance. Under this scenario, pests that can sense and avoid acquisition of lethal dosages of systemic insecticides within crop plants will have a selective advantage. This may lead to selection for insect behavioral avoidance, so that insects predominantly feed and oviposit on portions of crop plants with low concentration of systemic insecticide. Limited research has been published on the effect of environmental variables, including drought, on pesticide performance, but we present and discuss studies that support the hypothesis described above. In addition, we wish to highlight the importance of studying the many ways environmental factors can affect, directly and indirectly, both the performance of insecticides and the risk of target insect pests developing resistance.


Comparative evaluation of the tridimensional spine position measured with a new instrument (Vertebral Metrics) and an optoelectronic system of stereophotogrammetry.
We designed and built a non-invasive instrument, called Vertebral Metrics, to measure the x, y, and z positions of each spinous process of the spine on a standing position. In the present study, we perform a comparative evaluation of Vertebral Metrics, by comparing the results obtained from this instrument with those from a validated optoelectronic system of stereophotogrammetry, with 10 infrared cameras. The sample was composed of 11 women aged between 14 and 39 years. After marking the various points on the spinal column, from the first cervical vertebra to the first sacral vertebra, they were measured first with the new instrument (Vertebral Metrics) and then by means of the optoelectronics system. Afterwards, the results were subjected to a thorough comparison. The statistical comparison of the results was performed using an ANOVA model with three factors (Instrument, Subject, and Vertebra) for the intervertebral distance.


An endogenous tryptophan photo-product, FICZ, is potentially involved in photo-aging by reducing TGF-β-regulated collagen homeostasis.
Persistent ultraviolet (UV) radiation in the form of sunlight causes photo-aging of the skin by reducing the production of type I collagen, the major constituent of the extracellular matrix of the dermis. Transforming growth factor (TGF)-β transforms dermal fibroblasts into α2-smooth muscle actin (ACTA2)-expressing myofibroblasts. Myofibroblasts produce a precursor form of type I collagen, type I procollagen (collagen I), consisting of pro-alpha1 (produced by the COL1A1 gene) and pro-alpha2 chains (produced by the COL1A2 gene). Smad2/3 is a key downstream molecule of TGF-β signaling. The mechanisms through which UV inhibits collagen I synthesis are not fully understood. 6-Formylindolo[3,2-b]carbazole (FICZ) is an endogenous tryptophan photo-metabolite generated by UV irradiation. FICZ is well known as a high-affinity ligand for aryl hydrocarbon receptor (AHR). However, the physiological roles of FICZ in photo-aging have yet to be addressed. To evaluate the effects of FICZ on the TGF-β-mediated ACTA2 and collagen I expression in normal human dermal fibroblasts (NHDFs). Quantitative real-time polymerase chain reaction and western blot analysis were performed to determine the expression of ACTA2, COL1A1, and COL1A2 in NHDFs with or without FICZ and TGF-β. The phosphorylated Smad2/3 (pSmad2/3) protein levels in cytoplasmic or nuclear portions were investigated by western blot analysis. Immunofluorescence staining was conducted to evaluate pSmad2/3 localization, and F-actin staining with phalloidin was performed to visualize actin polymerization in myofibroblasts. The actions of FICZ on the TGF-β-mediated collagen I expression and nuclear translocation of pSmad2/3 were analyzed in the presence of selective AHR antagonists or in AHR-knockdown NHDFs. We found that FICZ significantly inhibited the TGF-β-induced upregulation of mRNA and protein levels of ACTA2 and collagen I and actin polymerization in myofibroblasts. FICZ did not disturb the phosphorylation of Smad2/3. Notably, FICZ reduced the expression of pSmad2/3 in the nucleus, while it increased that in the cytoplasm, suggesting that it inhibits the nuclear translocation of pSmad2/3 induced by TGF-β. The inhibitory actions of FICZ on the TGF-β-mediated collagen I expression and nuclear translocation of pSmad2/3 were independent of AHR signaling. Another endogenous AHR agonist, kynurenine, also inhibited the TGF-β-mediated ACTA2 and collagen I upregulation in NHDFs in an AHR-independent manner; however, its effects were insignificant in comparison with those of FICZ. These findings suggest that the endogenous photo-product FICZ may be a key chromophore that involves in photo-aging. Downregulation of FICZ signaling is thus a potential strategy to protect against photo-aging.


Susceptibility of Campylobacter jejuni and Yersinia enterocolitica to UV radiation.
Two enteric pathogens, Campylobacter jejuni and Yersinia enterocolitica serogroup O:3, together with Escherichia coli, were investigated for susceptibility to UV radiation at 254 nm. The UV dose required for a 3-log reduction (99.9% inactivation) of C. jejuni, Y. enterocolitica, and E. coli was 1.8, 2.7, and 5.0 mWs/cm2, respectively. Using E. coli as the basis for comparison, it appears that C. jejuni and Y. enterocolitica serogroup O:3 are more sensitive to UV than many of the pathogens associated with waterborne disease outbreaks and can be easily inactivated in most commercially available UV reactors. No association was found between the sensitivity of Y. enterocolitica to UV and the presence of a 40- to 50-megadalton virulence plasmid.


Benzodithiophenes potentiate differentiation of acute promyelocytic leukemia cells by lowering the threshold for ligand-mediated corepressor/coactivator exchange with retinoic acid receptor alpha and enhancing changes in all-trans-retinoic acid-regulated gene expression.
Differentiation induction is an effective therapy for acute promyelocytic leukemia (APL), which dramatically responds to all-trans-retinoic acid (ATRA). Recent studies have indicated that combinatorial use of retinoid and nonretinoid compounds, such as histone deacetylase inhibitors, arsenics, and PKA agonists, has higher therapeutic value in this disease and potentially in other malignancies. In a screen of 370 compounds, we identified benzodithiophene analogues as potent enhancers of ATRA-induced APL cell differentiation. These effects were not associated with changes in global histone acetylation and, for the most potent compounds, were exerted at very low nanomolar concentrations, and were paralleled by enhancement of some, but not all, ATRA-modulated gene expressions. Investigating the mechanism underlying the effects of these drugs on ATRA-induced APL cell differentiation, we have shown that benzodithiophenes enhance ATRA-mediated dissociation and association of corepressor N-CoR and coactivator p300 acetyltransferase, respectively, with retinoic acid receptor (RAR) alpha proteins. These data suggest that benzodithiophenes act at the level of receptor activation, possibly by affecting posttranslational modification of the receptor (and/or coregulators), thus leading to an enhancement in ATRA-mediated effects on gene expression and APL cell differentiation. Given the specificities of these low benzodithiophene concentrations for PML-RARalpha and RARalpha, these drugs may be useful for combinatorial differentiation therapy of APL and possibly other acute myelogenous leukemia subtypes in which the overall ATRA signaling is suppressed.


Lipopolysaccharide-induced gene expression in murine macrophages is enhanced by prior exposure to oxLDL.
Uptake of modified lipoproteins by macrophages results in the formation of foam cells. We investigated how foam cell formation affects the inflammatory response of macrophages. Murine bone marrow-derived macrophages were treated with oxidized LDL (oxLDL) to induce foam cell formation. Subsequently, the foam cells were activated with lipopolysaccharide (LPS), and the expression of lipid metabolism and inflammatory genes was analyzed. Furthermore, gene expression profiles of foam cells were analyzed using a microarray. We found that prior exposure to oxLDL resulted in enhanced LPS-induced tumor necrosis factor (TNF) and interleukin-6 (IL-6) gene expression, whereas the expression of the anti-inflammatory cytokine IL-10 and interferon-beta was decreased in foam cells. Also, LPS-induced cytokine secretion of TNF, IL-6, and IL-12 was enhanced, whereas secretion of IL-10 was strongly reduced after oxLDL preincubation. Microarray experiments showed that the overall inflammatory response induced by LPS was enhanced by oxLDL loading of the macrophages. Moreover, oxLDL loading was shown to result in increased nuclear factor-kappaB activation. In conclusion, our experiments show that the inflammatory response to LPS is enhanced by loading of macrophages with oxLDL. These data demonstrate that foam cell formation may augment the inflammatory response of macrophages during atherogenesis, possibly in an IL-10-dependent manner.


Increased circulating progranulin is not sufficient to induce cardiac dysfunction or supraventricular arrhythmia.
Atrial fibrillation (AF) is the most prevalent cardiac arrhythmia, and the incidence of new-onset AF has been increasing over the past two decades. Several factors contribute to the risk of developing AF including age, preexisting cardiovascular disease, chronic kidney disease, and obesity. Concurrent with the rise in AF, obesity has followed the same two-decade trend. The contribution of circulating proteins to obesity-related AF is of particular interest in the field. In this study, we investigated the effects of increased circulating levels of the glycoprotein progranulin on the development of supraventricular arrhythmias and changes to cardiac function. AAV8-mediated overexpression of full-length mouse progranulin was used to increase plasma protein levels and determine susceptibility to supraventricular arrhythmias and changes in cardiac structure and function. C57Bl/6N mice were subjected to increased circulating levels of progranulin for 20 weeks. Cardiac conduction was evaluated by surface ECG with and without isoproterenol challenge, and cardiac structure and function were measured by echocardiography after 20 weeks of circulating progranulin overexpression. Increased circulating levels of progranulin were maintained throughout the 20-week study. The cardiac structure and function remained unchanged in mice with increased circulating progranulin. ECG indices (P wave duration, P amplitude, QRS interval) were unaffected by increased progranulin levels and no arrhythmogenic events were observed following the isoproterenol challenge. In our model, increased levels of circulating progranulin were not sufficient to induce changes in cardiac structure and function or elicit ECG abnormalities suggestive of susceptibility to supraventricular arrhythmias.


Factors associated with interpersonal violence among children from public schools in Fortaleza, Ceará, Brazil.
The school has witnessed the growth of manifestations of interpersonal violence among children, and in this perspective, this paper examines the prevalence of interpersonal violence in the relationship between students and the associated factors. This is a cross-sectional study developed with 874 public school students aged 10 and 11 years conducted in 2013. A questionnaire implemented the collection of data that were submitted to bivariate analysis with statistical significance calculation between associations. The results show that the child-perpetrator (83.2%) is associated with the male gender (PR=1.08), as well as being beaten at home (PR=1.13) and having a family that encourages retaliation (PR=1.17). The child-victim (89.5%) is associated with the family that encourages retaliation (PR=1.05), participation in conflicts (93.6%) and age. The 10-year-old child is up to 3.0% more likely to participate in conflicts. The "family that encourages retaliation" is positively associated with the situations of assault, victim condition and participation in conflicts, which entails the reframing of these parental practices.


Effect of DISC1 SNPs on brain structure in healthy controls and patients with a history of psychosis.
Disrupted-in-Schizophrenia-1 (DISC1) has been suggested as a susceptibility locus for a broad spectrum of psychiatric disorders. Risk variants have been associated with brain structural changes, which overlap alterations reported in schizophrenia and bipolar disorder patients. We used genome-wide genotyping data for a Norwegian sample of healthy controls (n = 171) and patients with a history of psychosis (n = 184), to investigate 61 SNPs in the DISC1 region for putative association with structural magnetic resonance imaging (sMRI) measures (hippocampal volume; mean cortical thickness; and total surface area, as well as cortical thickness and area divided into four lobar measures). SNP rs821589 was associated with mean temporal and total brain cortical thickness in controls (P(adjusted) = 0.009 and 0.02, respectively), but not in patients. SNPs rs11122319 and rs1417584 were associated with mean temporal cortical thickness in patients (P(adjusted) = 0.04 and 0.03, respectively), but not in controls, and both SNPs have previously been highly associated with DISC1 gene expression. There were significant genotype × case-control interactions. There was no significant association between SNPs and cortical area or hippocampal volume in controls, or with any of the structural measures in cases, after correction for multiple comparisons. In conclusion, DISC1 SNPs might impact brain structural variation, possibly differently in psychosis patients versus controls, but independent replication will be needed to confirm our findings.


Biomechanics of the Sacroiliac Joint: Anatomy, Function, Biomechanics, Sexual Dimorphism, and Causes of Pain.
The sacroiliac joints (SIJs), the largest axial joints in the body, sit in between the sacrum and pelvic bones on either side. They connect the spine to the pelvis and thus facilitate load transfer from the lumbar spine to the lower extremities. The majority of low back pain (LBP) is perceived to originate from the lumbar spine; however, another likely source of LBP that is mostly overlooked is the SIJ. This study (Parts I and II) aims to evaluate the clinical and biomechanical literature to understand the anatomy, biomechanics, sexual dimorphism, and causes and mechanics of pain of the SIJ leading to conservative and surgical treatment options using instrumentation. Part II concludes with the mechanics of the devices used in minimal surgical procedures for the SIJ. A thorough review of the literature was performed to analyze studies related to normal SIJ mechanics, as well as the effects of sex and pain on SIJ mechanics. A total of 65 studies were selected related to anatomy, biomechanical function of the SIJ, and structures that surround the joints. These studies discussed the effects of various parameters, gender, and existence of common physiological disorders on the biomechanics of the SIJ. The SIJ lies between the sacrum and the ilium and connects the spine to the pelvic bones. The SIJ transfers large bending moments and compression loads to lower extremities. However, the joint does not have as much stability of its own against the shear loads but resists shear due the tight wedging of the sacrum between hip bones on either side and the band of ligaments spanning the sacrum and the hip bones. Due to these, sacrum does not exhibit much motion with respect to the ilium. The SIJ range of motion in flexion-extension is about 3°, followed by axial rotation (about 1.5°), and lateral bending (about 0.8°). The sacrum of the female pelvis is wider, more uneven, less curved, and more backward tilted, compared to the male sacrum. Moreover, women exhibit higher mobility, stresses/loads, and pelvis ligament strains compared to male SIJs. Sacroiliac pain can be due to, but not limited to, hypo- or hypermobility, extraneous compression or shearing forces, micro- or macro-fractures, soft tissue injury, inflammation, pregnancy, adjacent segment disease, leg length discrepancy, and prior lumbar fusion. These effects are well discussed in this review. This review leads to Part II, in which the literature on mechanics of the treatment options is reviewed and synthesized.


Cross-cultural adaptation, reliability and construct validity of the Tampa scale for kinesiophobia for temporomandibular disorders (TSK/TMD-Br) into Brazilian Portuguese.
Fear of movement (kinesiophobia) seems to play an important role in the development of chronic pain. However, for temporomandibular disorders (TMD), there is a scarcity of studies about this topic. The Tampa Scale for Kinesiophobia for TMD (TSK/TMD) is the most widely used instrument to measure fear of movement and it is not available in Brazilian Portuguese. The purpose of this study was to culturally adapt the TSK/TMD to Brazilian Portuguese and to assess its psychometric properties regarding internal consistency, reliability, and construct and structural validity. A total of 100 female patients with chronic TMD participated in the validation process of the TSK/TMD-Br. The intraclass correlation coefficient (ICC) was used for statistical analysis of reliability (test-retest), Cronbach's alpha for internal consistency, Spearman's rank correlation for construct validity and confirmatory factor analysis (CFA) for structural validity. CFA endorsed the pre-specified model with two domains and 12-items (Activity Avoidance - AA/Somatic Focus - SF) and all items obtained a loading factor greater than 0·4. Acceptable levels of reliability were found (ICC > 0·75) for all questions and domains of the TSK/TMD-Br. For internal consistency, Cronbach's α of 0·78 for both domains were found. Moderate correlations (0·40 < r < 0.60) were observed for 84% of the analyses conducted between TSK/TMD-Br scores versus catastrophising, depression and jaw functional limitation. TSK/TMD-Br 12 items and two-factor demonstrated sound psychometric properties (transcultural validity, reliability, internal consistency and structural validity). In such a way, the instrument can be used in clinical settings and for research purposes.


SMC1 promotes proliferation and inhibits apoptosis through the NF‑κB signaling pathway in colorectal cancer.
Colorectal cancer (CRC) is the third most common cancer and a leading cause of cancer‑associated mortality globally. Increasing evidence has indicated that structural maintenance of chromosomes 1 (SMC1) may serve an important role in solid tumors as a tumor enhancer. In the present study, it was identified via reverse transcription‑quantitative PCR and western blot analyses that expression of SMC1 was significantly upregulated in CRC cell lines (SW480, HCT‑116 and SW620) compared with NCM460 normal epithelial cells. To determine the potential involvement of SMC1 in the malignant phenotypes of CRC cells, SMC1 was knocked down in SW620 cells and SMC1 was overexpressed in SW480 cells in vitro. As a result, cell viability, proliferation, invasion and migration were suppressed in transduced SW620 cells but enhanced in SW480 cells, as determined using MTT, colony formation and Transwell assays; conversely, flow cytometric analysis revealed that cell apoptosis was increased in SW620 cells and inhibited in SW480 cells following lentiviral infection. In addition, an in vivo mouse xenograft model revealed that knocking down SMC1 suppressed tumor growth and increased apoptosis; however, overexpressing SMC1 enhanced tumor growth and suppressed apoptosis. Further experiments demonstrated that the role of SMC1 on CRC may involve downregulation of the NF‑κB‑associated signaling pathway. Finally, the present data from clinical CRC tumor samples showed that increased expression of SMC1 was significantly associated with distant metastasis, higher TNM stage, primary tumor size, lymph node metastasis and worse overall survival. Collectively, the present results suggested that SMC1 served an important role in the development of CRC and may be a predictive prognostic biomarker in patients with CRC.


Evaluation of Pharyngeal Airway Dimensions and Hyoid Bone Position in Children After Adenoidectomy or Adenotonsillectomy: A Cephalometric Study.
Background. The study aimed to compare the airway morphology and hyoid bone position in children undergoing adenoidectomy or adenotonsillectomy with healthy individuals with no skeletal problems in similar age and development period. Methods. The subjects in the study were divided into three groups. These groups were defined as those having undergone adenoidectomy (53 children), those having undergone adenotonsillectomy (48 children), and the systemically healthy control group (63 children). Seventeen pharyngeal airway, nine hyoid, and four area measurements were used in the cephalometric radiographs of the children in all the groups. One-way analysis of variance was used to evaluate the children in each group. In addition, Tukey tests were used for a bilateral comparison of significant parameters. The results were considered statistically significant at P<0.05. Results. The mean age of 164 patients in the study group was 12.76±2.85 years. The vertical airway length significantly decreased in the adenotonsillectomy group compared to other groups, while the epiglottic pharyngeal length significantly increased in the former than in the latter (P<0.05). The area measurements showed that although the oropharynx area significantly increased in the adenotonsillectomy group compared to other groups, the hypopharynx and total area measurements were significantly different compared with the control group (P<0.05). No statistically significant difference was found between the three groups in all measurements of the hyoid bone position (P>0.05). Conclusion. The study showed that adenotonsillectomy operations caused more increases in the oropharynx and hypopharynx parts of the pharyngeal airway. Adenoidectomy and adenotonsillectomy operations did not contribute significantly to the hyoid bone position.


The transvestibular approach: a new horizon in rhinoplasty.
The lower lateral cartilage has intricate anatomical elements that define shape, projection, and relations with other nasal tip structures. Good exposure of the lower lateral cartilages is an essential step in rhinoplasty. Conservative surgical techniques are essential to get natural long-term results. Different endonasal techniques have limitations in visualizing the lateral and intermediate crura, predisposing patients to asymmetries in reduction and rearrangement. In this article, a new endonasal rhinoplasty approach is described. Using a marginal incision, the vestibular skin is elevated and the endonasal surface of the lower lateral cartilage is exposed, permitting precise reduction, rearrangement, and placement of interdomal suspension sutures and lateral crural spanning sutures. This article discusses the surgical steps and results of my experience using this approach. This transvestibular approach is a new, dependable, and simple method that should allow rhinoplastic surgeons to perform this operation with predictable results and limited variables.


Functional and historical determinants of shape in the scapula of Xenarthran mammals: evolution of a complex morphological structure.
The mammalian scapula is a complex morphological structure, composed of two ossification plates that fuse into a single structure. Most studies on morphological differentiation in the scapula have considered it to be a simple, spatially integrated structure, primarily influenced by the important locomotor function presented by this element. We used recently developed geometric morphometric techniques to test and quantify functional and phylogenetic influences on scapular shape variation in fossil and extant xenarthran mammals. The order Xenarthra is well represented in the fossil record and presents a stable phylogenetic hypothesis for its genealogical history. In addition, its species present a large variety of locomotor habits. Our results show that approximately half of the shape variation in the scapula is due to phylogenetic heritage. This is contrary to the view that the scapula is influenced only by functional demands. There are large-scale shape transformations that provide biomechanical adaptation for the several habits (arboreality, terrestriality, and digging), and small scale-shape transformations (mostly related to the coracoid process) that are not influenced by function. A nonlinear relationship between morphometric and phylogenetic distances indicates the presence of a complex mixture of evolutionary processes acting on shape differentiation of the scapula.


Synthesis and anti-HIV-1 activity of thio analogues of dihydroalkoxybenzyloxopyrimidines.
Various thio analogues of dihydroalkoxybenzyloxopyrimidines (DABOs), a new class of non-nucleoside reverse transcriptase inhibitors, were found to selectively inhibit the HIV-1 multiplication in vitro. Among the C-5 H-substituted 6-benzyl-3,4-dihydro-4-oxopyrimidines, the introduction of alkylthio or cycloalkylthio substituents at C-2 of the pyrimidine ring led to derivatives (S-DABOs) which were up to 10-fold more potent than the alkyloxy or cycloalkyloxy counterparts. The further introduction of a methyl group at the 3'-position of the benzyl portion of 2-(alkylthio)-6-benzyluracils reduced the cytotoxicity leading to more selective compounds. Among C-5 methyl-substituted S-DABOs, numerous derivatives showed EC50 values as low as 0.6 microM and lacked cytotoxicity at doses as high as 300 microM. In the C-5 double methyl-substituted series, a more pronounced cytotoxicity was observed and the further introduction of a methyl at the 3'-position in the benzylidene group resulted in total loss of antiviral activity. S-DABOs, namely 2-(alkylthio)-6-benzyl-3,4-dihydro-4-oxopyrimidines, were synthesized by reacting proper methyl (phenylacetyl)acetates or their 2-methyl compounds with thiourea to afford 6-benzyl-4-oxo-1,2,3,4-tetrahydro-2-thiaoxopyrimidines or the related 5-methyl derivatives. Treatment of the latter derivatives with alkyl or cycloalkyl halides in alkaline medium gave the required title compounds.


Nicotinic acetylcholine receptors in glucose homeostasis: the acute hyperglycemic and chronic insulin-sensitive effects of nicotine suggest dual opposing roles of the receptors in male mice.
Cigarette smoking causes insulin resistance. However, nicotine induces anti-inflammation and improves glucose tolerance in insulin-resistant animal models. Here, we determined the effects of nicotine on glucose metabolism in insulin-sensitive C57BL/J6 mice. Acute nicotine administration (30 min) caused fasting hyperglycemia and lowered insulin sensitivity acutely, which depended on the activation of nicotinic-acetylcholine receptors (nAChRs) and correlated with increased catecholamine secretion, nitric oxide (NO) production, and glycogenolysis. Chlorisondamine, an inhibitor of nAChRs, reduced acute nicotine-induced hyperglycemia. qRT-PCR analysis revealed that the liver and muscle express predominantly β4 > α10 > α3 > α7 and β4 > α10 > β1 > α1 mRNA for nAChR subunits respectively, whereas the adrenal gland expresses β4 > α3 > α7 > α10 mRNA. Chronic nicotine treatment significantly suppressed expression of α3-nAChR (predominant peripheral α-subunit) in liver. Whereas acute nicotine treatment raised plasma norepinephrine (NE) and epinephrine (Epi) levels, chronic nicotine exposure raised only Epi. Acute nicotine treatment raised both basal and glucose-stimulated insulin secretion (GSIS). After chronic nicotine treatment, basal insulin level was elevated, but GSIS after acute saline or nicotine treatment was blunted. Chronic nicotine exposure caused an increased buildup of NO in plasma and liver, leading to decreased glycogen storage, along with a concomitant suppression of Pepck and G6Pase mRNA, thus preventing hyperglycemia. The insulin-sensitizing effect of chronic nicotine was independent of weight loss. Chronic nicotine treatment enhanced PI-3-kinase activities and increased Akt and glycogen synthase kinase (GSK)-3β phosphorylation in an nAChR-dependent manner coupled with decreased cAMP response element-binding protein (CREB) phosphorylation. The latter effects caused suppression of Pepck and G6Pase gene expression. Thus, nicotine causes both insulin resistance and insulin sensitivity depending on the duration of the treatment.


Nutritional advice in pregnancy.
To assess the effects of advising pregnant women to increase their energy and protein intakes on those intakes, on gestational weight gain, and on the outcome of pregnancy. The register of clinical trials maintained and updated by the Cochrane Pregnancy and Childbirth Group. All acceptably controlled comparisons of nutritional advice, whether administered on a one-to-one basis or to groups of women. Data were extracted by the author from published reports, and supplemented by additional information from trialists contacted by the author. Four trials involving 1108 women were included. Advice to increase energy and protein intakes seems to be successful in achieving those goals, but the increases are lower than those reported in trials of actual protein/energy supplementation. Data concerning effects on pregnancy outcome are available only from one trial, and, given the fact that its analysis was based on individual women despite randomization by clinic, the calculated confidence intervals are undoubtedly too narrow. Moreover, the 'significant' reduction in preterm birth associated with advice is not consistent with the total absence of effect on mean gestational age. One trial found no reduction in the incidence of pre-eclampsia. No data have been reported on potential adverse effects that might accompany increased fetal size, such as an increased risk of prolonged labour or Caesarean section. Nutritional advice appears effective in increasing pregnant women's energy and protein intakes, but the implications for fetal, infant, or maternal health cannot be judged from the available trials. Given the rather modest health benefits demonstrated with actual protein/energy supplementation (see the Cochrane review of 'Balanced protein/energy supplementation in pregnancy'), however, the provision of such advice is unlikely to be of major importance.


"Robbery by hypnosis" in Italy: a psycho-criminological analysis of the phenomenon based on 20 years of newspaper articles (1988-2007).
Shocking news of robberies committed using hypnosis on bank cashiers, salespeople, or passers-by has sporadically been reported by the media in countries around the world. The first reported episode in Italy dates to the 1950s. Although the phenomenon has been reported in the papers more frequently in recent years, no objective analysis of it has been published in the scientific literature. This paper analyzes 106 episodes recorded in Italy between 1988 and 2007, identified by a systematic review of the online and printed archives of Italian national and local dailies and of the database of the country's principal press agency. When they are analyzed from a psychological and criminological standpoint, there is no evidence to support any real use of hypnotic methods in the episodes described.


Pain, Quality of Life, and Clinical Outcomes after Robotic Lobectomy.
Background To evaluate pulmonary function, pain, and quality of life at midterm after robotic lobectomy performed in a single institution. Methods Sixty-five consecutive patients underwent robotic thoracic surgery over 32 months using a complete four-arm portal technique. Sixty-one patients underwent lobectomies predominantly for stage I non-small cell lung cancer. Pulmonary function tests were repeated at midterm follow-up. Pain and quality of life were evaluated during the follow-up on a subgroup of 39 patients, excluding the learning period. Results At a mean of 7-month follow-up, there was no significant difference in preoperative and midterm postoperative pulmonary function. A total of 62.5% of the patients reported a variable intensity of discomfort or pain at the surgical site, with a mean pain intensity score of 2.1 ± 1.4. Mean pain interference score were weak (1.8 ± 1.9), with patients with moderate pain reporting significantly higher pain interference scores than those with mild pain (p = 0.0025). Only one patient suffered from neuropathic-like pain. Quality of life was globally favorable and related to the pain level, with a significant interference on the physical component. Conclusion Robotic lobectomy does not appear to have an impact on midterm pulmonary function. Persistent postoperative pain is mild, nonneuropathic-like, with weak interference on daily activities. Quality of life is satisfactory but related to the pain level.


ARRDC5 deficiency impairs spermatogenesis by affecting SUN5 and NDC1.
Sperm with normal morphology and motility are essential for successful fertilization, and the strong attachment of the sperm head-tail coupling apparatus to the nuclear envelope during spermatogenesis is required to ensure the integrity of sperm for capacitation and fertilization. Here, we report that Arrdc5 is associated with spermatogenesis. The Arrdc5 knockout mouse model showed male infertility characterized by a high bent-head rate and reduced motility in sperm, which led to capacitation defects and subsequent fertilization failure. Through mass spectrometry, we found that ARRDC5 affects spermatogenesis by affecting NDC1 and SUN5. We further found that ARRDC5 might affect the vesicle-trafficking protein SEC22A-mediated transport and localization of NDC1, SUN5 and other head-tail coupling apparatus-related proteins that are responsible for initiating the attachment of the sperm head and tail. We finally performed intracytoplasmic sperm injection as a way to explore therapeutic strategies. Our findings demonstrate the essential role and the underlying molecular mechanism of ARRDC5 in anchoring the sperm head to the tail during spermatogenesis.


[Drug prescription for pregnant women: a pharmacoepidemiological study].
Prescription drug use by pregnant women should be viewed as a public health issue, since there are numerous gaps in knowledge on the consequences for both the mother and the fetus. Pharmacoepidemiological studies can help minimize the inherent risks in drug treatment by establishing a profile of drug consumption during pregnancy, providing an evaluation of this service, and identifying intervention measures. The purpose of this study was to establish a prescription profile for pregnant women treated under the National Health System (SUS) in Piracicaba, São Paulo State, Brazil, using prescription indicators recommended by the World Health Organization and the U.S. Food and Drug Administration classification of medicines according to risk to the fetus. According to the current study, during prenatal consultations 44.7% of the women received drug prescriptions, and the most widely prescribed group of drugs were those acting on the hematopoietic system (34.9%). Of this total, 26.0% of the drugs were included in fetal risk category C, 1.5% in category D, and 1.5% in category E. These data point to the medicalization of pregnancy and the need for intervention measures aimed at rational prescription drug use during the prenatal period.


Antioxidant and Immunostimulatory Activities of a Submerged Culture of Cordyceps sinensis Using Spent Coffee.
Spent coffee grounds (SCG) are inexpensive materials that have been used as a source of antioxidants and polysaccharides with immunostimulatory activity. In this study, we performed a microbial fermentation of SCG using Cordyceps sinensis and investigated the radical scavenging and immunostimulatory activity of fermented SCG. SCG fermentation using C. sinensis was performed at 25 °C for 8 d. The polyphenol content of the fermented SCG increased from 1022.4 to 1562.0 μg/mL. The glucosamine content of the mycelia also continuously increased during fermentation. The main polyphenol compounds of fermented SCG were chlorogenic acid and p-coumaric acid, which were increased by fermentation. Fermented SCG also showed significantly higher content of chlorogenic acid isomers than unfermented SCG. The fermented SCG exhibited significantly higher 2,2-diphenyl-2-picrylhydrazyl hydrate (half maximal inhibitory concentration: IC50, 0.37 mg/mL) and 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (IC50, 0.93 mg/mL) radical scavenging activities than those of the control (0.54 mg/mL and 1.20 mg/mL, respectively; p < 0.05). The fermented SCG stimulated macrophages and promoted the production of various immunostimulatory cytokines (IL-12, IL-6, and TNF-α) compared to control; therefore, microbial fermentation of SCG using C. sinensis is an effective means of generating antioxidant and immunostimulatory materials.


The carbohydrate at asparagine 386 on HIV-1 gp120 is not essential for protein folding and function but is involved in immune evasion.
The HIV-1 envelope glycoprotein gp120, which mediates viral attachment to target cells, consists for approximately 50% of sugar, but the role of the individual sugar chains in various aspects of gp120 folding and function is poorly understood. Here we studied the role of the carbohydrate at position 386. We identified a virus variant that had lost the 386 glycan in an evolution study of a mutant virus lacking the disulfide bond at the base of the V4 domain. The 386 carbohydrate was not essential for folding of wt gp120. However, its removal improved folding of a gp120 variant lacking the 385-418 disulfide bond, suggesting that it plays an auxiliary role in protein folding in the presence of this disulfide bond. The 386 carbohydrate was not critical for gp120 binding to dendritic cells (DC) and DC-mediated HIV-1 transmission to T cells. In accordance with previous reports, we found that N386 was involved in binding of the mannose-dependent neutralizing antibody 2G12. Interestingly, in the presence of specific substitutions elsewhere in gp120, removal of N386 did not result in abrogation of 2G12 binding, implying that the contribution of N386 is context dependent. Neutralization by soluble CD4 and the neutralizing CD4 binding site (CD4BS) antibody b12 was significantly enhanced in the absence of the 386 sugar, indicating that this glycan protects the CD4BS against antibodies. The carbohydrate at position 386 is not essential for protein folding and function, but is involved in the protection of the CD4BS from antibodies. Removal of this sugar in the context of trimeric Env immunogens may therefore improve the elicitation of neutralizing CD4BS antibodies.


Functional Analysis of Cellulose and Xyloglucan in the Walls of Stomatal Guard Cells of Arabidopsis.
Stomatal guard cells are pairs of specialized epidermal cells that control water and CO2 exchange between the plant and the environment. To fulfill the functions of stomatal opening and closure that are driven by changes in turgor pressure, guard cell walls must be both strong and flexible, but how the structure and dynamics of guard cell walls enable stomatal function remains poorly understood. To address this question, we applied cell biological and genetic analyses to investigate guard cell walls and their relationship to stomatal function in Arabidopsis (Arabidopsis thaliana). Using live-cell spinning disk confocal microscopy, we measured the motility of cellulose synthase (CESA)-containing complexes labeled by green fluorescent protein (GFP)-CESA3 and observed a reduced proportion of GFP-CESA3 particles colocalizing with microtubules upon stomatal closure. Imaging cellulose organization in guard cells revealed a relatively uniform distribution of cellulose in the open state and a more fibrillar pattern in the closed state, indicating that cellulose microfibrils undergo dynamic reorganization during stomatal movements. In cesa3(je5) mutants defective in cellulose synthesis and xxt1 xxt2 mutants lacking the hemicellulose xyloglucan, stomatal apertures, changes in guard cell length, and cellulose reorganization were aberrant during fusicoccin-induced stomatal opening or abscisic acid-induced stomatal closure, indicating that sufficient cellulose and xyloglucan are required for normal guard cell dynamics. Together, these results provide new insights into how guard cell walls allow stomata to function as responsive mediators of gas exchange at the plant surface.


Bioactivation of 2-nitrofluorene to reactive intermediates that bind covalently to DNA, RNA and protein in vitro and in vivo in the rat.
The activation of 2-nitrofluorene (2-NF) to reactive intermediates that bind covalently to DNA, RNA and protein has been investigated both in vitro and in the rat in vivo. In vitro, such binding was catalyzed by the hepatic microsomal fraction, was NADPH dependent and could be inhibited by SKF 525A, an inhibitor of cytochrome P450. The generation of reactive intermediates therefore is most likely catalyzed by cytochrome P450. Covalent binding of 2-NF could not be prevented by glutathione, N-acetylcysteine and other thiol-containing compounds. It could be partially prevented by guanosine, presumably because it traps the reactive intermediate(s). Under normal oxygenation conditions 2-NF was also covalently bound in freshly isolated hepatocytes; pretreatment of rats with an inducer of cytochrome P450, Aroclor 1254, gave rise to a higher rate of covalent binding in hepatocytes. Covalent binding of 2-NF to cellular macromolecules also occurred in vivo, both after oral and i.v. administration. Pentachlorophenol, a selective sulfation inhibitor, did not influence the covalent binding of 2-NF; therefore, the reactive intermediate is not formed by sulfation of N-hydroxy-2-acetylaminofluorene, which could be a metabolite of 2-NF. It is concluded that the reactive intermediates most likely can be formed from 2-NF by the cytochrome P450 enzyme system.


Human herpesvirus diversity is altered in HLA class I binding peptides.
Herpesviruses are ubiquitous, genetically diverse DNA viruses, with long-term presence in humans associated with infrequent but significant pathology. Human leukocyte antigen (HLA) class I presents intracellularly derived peptide fragments from infected tissue cells to CD8+ T and natural killer cells, thereby directing antiviral immunity. Allotypes of highly polymorphic HLA class I are distinguished by their peptide binding repertoires. Because this HLA class I variation is a major determinant of herpesvirus disease, we examined if sequence diversity of virus proteins reflects evasion of HLA presentation. Using population genomic data from Epstein–Barr virus (EBV), human cytomegalovirus (HCMV), and Varicella–Zoster virus, we tested whether diversity differed between the regions of herpesvirus proteins that can be recognized, or not, by HLA class I. Herpesviruses exhibit lytic and latent infection stages, with the latter better enabling immune evasion. Whereas HLA binding peptides of lytic proteins are conserved, we found that EBV and HCMV proteins expressed during latency have increased peptide sequence diversity. Similarly, latent, but not lytic, herpesvirus proteins have greater population structure in HLA binding than nonbinding peptides. Finally, we found patterns consistent with EBV adaption to the local HLA environment, with less efficient recognition of EBV isolates by high-frequency HLA class I allotypes. Here, the frequency of CD8+ T cell epitopes inversely correlated with the frequency of HLA class I recognition. Previous analyses have shown that pathogen-mediated natural selection maintains exceptional polymorphism in HLA residues that determine peptide recognition. Here, we show that HLA class I peptide recognition impacts diversity of globally widespread pathogens.


Online quench-flow electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry for elucidating kinetic and chemical enzymatic reaction mechanisms.
We have developed an automated quench-flow microreactor which interfaces directly to an electrospray ionization (ESI) mass spectrometer. We have used this device in conjunction with ESI Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS) to demonstrate the potential of this approach for studying the mechanistic details of enzyme reactions. For the model system chosen to test this device, namely, the pre-steady-state hydrolysis of p-nitrophenyl acetate by the enzyme chymotrypsin, the kinetic parameters obtained are in good agreement with those in the literature. To our knowledge, this is the first reported use of online quench-flow coupled with FTICR MS. Furthermore, we have exploited the power of FTICR MS to interrogate the quenched covalently bound enzyme intermediate using top-down fragmentation. The accurate mass capabilities of FTICR MS permitted the nature of the intermediate to be assigned with high confidence. Electron capture dissociation (ECD) fragmentation allowed us to locate the intermediate to a five amino acid section of the protein--which includes the known catalytic residue, Ser(195). This experimental approach, which uniquely can provide both kinetic and chemical details of enzyme mechanisms, is a potentially powerful tool for studies of enzyme catalysis.


Cell biological characterization of a multidomain adaptor protein, ArgBP2, in epithelial NMuMG cells, and identification of a novel short isoform.
ArgBP2 is a member of the SoHo (sorbin-homology) family of adaptor proteins believed to play roles in cell adhesion, cytoskeletal organization, and signaling. We show here a novel splicing isoform of ArgBP2, i.e., ArgBP2™, composed of only three SH3 (src-homology 3) domains and structurally similar to vinexinß. We then characterized the biochemical and cell biological properties of ArgBP2 to compare these with vinexin. Similar to vinexin, ArgBP2 was enriched at focal adhesions in REF52 fibroblast cells and induced anchorage-dependent extracellular signal-regulated kinase activation in NIH3T3 fibroblast cells. In epithelial NMuMG cells, immunofluorescence analyses revealed localization of ArgBP2 at tight junctions (TJs), whereas vinexin was distributed in cytoplasm as well as cell-cell boundaries. During TJ formation, recruitment of ZO-1 to TJs was followed by ArgBP2. Based on mutation analyses, a second SH3 domain was found to be important for ArgBP2 localization to the cell-cell contact sites. These data suggest some role of ArgBP2 in NMuMG cells at TJs that may be distinct from the function of vinexin.


Haloperidol augmentation of fluvoxamine in skin picking disorder: a case report.
Compulsive skin picking, being part of the broader category of impulse control disorders, is considered a residual diagnosis in the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision. It is characterized by excessive scratching or picking of normal skin, or skin with minor surface irregularities, and occurs in 2% of patients attending dermatology clinics. Despite the clinical relevance of this disorder, no clear guidelines are available yet; clinical management is, therefore, compromised and the day-to-day clinical practice is burdened by difficulties. Studies on selective serotonin reuptake inhibitors and anti-epileptic drugs have provided limited results. The association between anti-depressants and anti-epileptics has been found to be beneficial in some impulse control disorders, but in skin picking no previous studies have been conducted on this pharmacological approach. There are very few reports on the efficacy of anti-psychotics in skin picking. The therapeutic path described in this case report produced good results for a 59-year-old Caucasian woman. The first therapeutic approach, with fluvoxamine and oxcarbazepine was partially effective; then, the suspension of oxcarbazepine and haloperidol augmentation of fluvoxamine were adopted. After 10 weeks, a significant improvement of the disease was observed: the clinical picture and the associated symptoms were nearly solved. To the best of our knowledge, this is the first article reporting the association of fluvoxamine and haloperidol in skin picking disorder. It might be useful to perform further research regarding the treatment of skin picking disorder: in clinical practice, several variables might limit the choice of certain drugs. Therefore, it would be useful for the clinician to be aware of other therapeutic options.


Aluminum Ingestion Promotes Colorectal Hypersensitivity in Rodents.
Irritable bowel syndrome (IBS) is a multifactorial disease arising from a complex interplay between genetic predisposition and environmental influences. To date, environmental triggers are not well known. Aluminum is commonly present in food, notably by its use as food additive. We investigated the effects of aluminum ingestion in rodent models of visceral hypersensitivity, and the mechanisms involved. Visceral hypersensitivity was recorded by colorectal distension in rats administered with oral low doses of aluminum. Inflammation was analyzed in the colon of aluminum-treated rats by quantitative PCR for cytokine expression and by immunohistochemistry for immune cells quantification. Involvement of mast cells in the aluminum-induced hypersensitivity was determined by cromoglycate administration of rats and in mast cell-deficient mice (KitW-sh/W-sh). Proteinase-activated receptor-2 (PAR2) activation in response to aluminum was evaluated and its implication in aluminum-induced hypersensitivity was assessed in PAR2 knockout mice. Orally administered low-dose aluminum induced visceral hypersensitivity in rats and mice. Visceral pain induced by aluminum persisted over time even after cessation of treatment, reappeared and was amplified when treatment resumed. As observed in humans, female animals were more sensitive than males. Major mediators of nociception were up-regulated in the colon by aluminum. Activation of mast cells and PAR2 were required for aluminum-induced hypersensitivity. These findings indicate that oral exposure to aluminum at human dietary level reproduces clinical and molecular features of IBS, highlighting a new pathway of prevention and treatment of visceral pain in some susceptible patients.


Synergy between the NAMPT inhibitor GMX1777(8) and pemetrexed in non-small cell lung cancer cells is mediated by PARP activation and enhanced NAD consumption.
GMX1778 and its prodrug GMX1777 represent a new class of cancer drugs that targets nicotinamide phosphoribosyltransferase (NAMPT) as a new strategy to interfere with biosynthesis of the key enzymatic cofactor NAD, which is critical for a number of cell functions, including DNA repair. Using a genome-wide synthetic lethal siRNA screen, we identified the folate pathway-related genes, deoxyuridine triphosphatase and dihydrofolate reductase, the silencing of which sensitized non-small cell lung carcinoma (NSCLC) cells to the cytotoxic effects of GMX. Pemetrexed is an inhibitor of dihydrofolate reductase currently used to treat patients with nonsquamous NSCLC. We found that combining pemetrexed with GMX1777 produced a synergistic therapeutic benefit in A549 and H1299 NSCLC cells in vitro and in a mouse A549 xenograft model of lung cancer. Pemetrexed is known to activate PARPs, thereby accelerating NAD consumption. Genetic or pharmacologic blockade of PARP activity inhibited this effect, impairing cell death by pemetrexed either alone or in combination with GMX1777. Conversely, inhibiting the base excision repair pathway accentuated NAD decline in response to GMX and the cytotoxicity of both agents either alone or in combination. These findings provide a mechanistic rationale for combining GMX1777 with pemetrexed as an effective new therapeutic strategy to treat nonsquamous NSCLC.


Deazaflavin cofactor boosts earthworms Henlea bioluminescence.
The bioluminescence of Siberian earthworms Henlea sp. was found to be enhanced by two low molecular weight activators, termed ActH and ActS, found in the hot extracts. The fluorescence emission maximum of the activators matches the bioluminescence spectrum that peaks at 464 nm. We purified 4.3 and 8.8 micrograms of ActH and ActS from 200 worms and explored them using orbitrap HRMS with deep fragmentation and 1D/2D NMR equipped with cryoprobes. Their chemical structures were ascertained using chemical shift prediction services, structure elucidation software and database searches. ActH was identified as the riboflavin analoge archaeal cofactor F0, namely 7,8-didemethyl-8-hydroxy-5-deazariboflavin. ActS is a novel compound, namely ActH sulfated at the 3' ribityl hydroxyl. We designed and implemented a new four step synthesis strategy forActH that outperformed previous synthetic approaches. The synthetic ActH was identical to the natural one and activated Henlea sp. bioluminescence. The bioluminescence enhancement factor X was measured at different ActH concentrations and the Michaelis constant Km = 0.22 ± 0.01 μM was obtained by nonlinear regression. At an excess of synthetic ActH, the factor X was saturated at Xmax = 33.3 ± 0.5, thus opening an avenue to further characterisation of the Henlea sp. bioluminescence system. ActH did not produce bioluminescence without the luciferin with an as yet unknown chemical structure. We propose that ActH and the novel sulfated deazariboflavin ActS either emit the light of the Henlea sp. bioluminescence and/or accept hydride(s) donor upon luciferin oxidation.


High sodium and low potassium intake among Italian children: relationship with age, body mass and blood pressure.
Hypertension is the leading cause of death in developed countries and reduction of salt intake is recommended as a key preventive measure. To assess the dietary sodium and potassium intakes in a national sample of Italian children and adolescents and to examine their relationships with BMI and blood pressure (BP) in the framework of the MINISAL survey, a program supported by the Italian Ministry of Health. The study population included 1424 healthy subjects (766 boys, 658 girls) aged 6-18 years (mean age: 10.1±2.9) who were consecutively recruited in participating National Health Service centers in 10 Italian regions. Electrolyte intake was estimated from 24 hour urine collections tested for completeness by the concomitant measurement of creatinine content. Anthropometric indices and BP were measured with standardized procedures. The average estimated sodium intake was 129 mmol (7.4 g of salt) per day among boys and 117 mmol (6.7 g of salt) among girls. Ninety-three percent of the boys and 89% of the girls had a consumption higher than the recommended age-specific standard dietary target. The estimated average daily potassium intakes were 39 mmol (1.53 g) and 36 mmol (1.40 g), respectively, over 96% of the boys and 98% of the girls having a potassium intake lower than the recommended adequate intake. The mean sodium/potassium ratio was similar among boys and girls (3.5 and 3.4, respectively) and over 3-fold greater than the desirable level. Sodium intake was directly related to age, body mass and BP in the whole population. The Italian pediatric population is characterized by excessive sodium and deficient potassium intake. These data suggest that future campaigns should focus on children and adolescents as a major target in the framework of a population strategy of cardiovascular prevention.


Employers.
By shifting employees into managed care, demanding accountability from providers, and negotiating in groups, employers have helped lower their health care costs.


Complete genome sequence of Streptomyces strain VNUA116, a potential biocontrol against phytopathogenic fusarium wilt fungus Fusarium oxysporum f. sp. cubense.
Here, we present the whole-genome sequence of Streptomyces strain VNUA116 was obtained by combining sequencing data from both PacBio RS II and DNBseq platforms. The complete circular genome is 8,306,919 bp with a GC content of 72.49%.


Enhanced proteolysis of plasma von Willebrand factor in thrombotic thrombocytopenic purpura and the hemolytic uremic syndrome.
To examine whether enhanced in vivo proteolysis of von Willebrand factor (vWF) would account for the reported loss of larger multimers in acute thrombotic thrombocytopenic purpura/hemolytic uremic syndrome (TTP/HUS), we studied eight patients with acute TTP/HUS whose blood samples were collected into an anticoagulant containing a cocktail of protease inhibitors to impede in vitro proteolysis. In all, enhanced proteolytic degradation of vWF was expressed as a relative decrease in the intact 225-Kd subunit of vWF and a relative increase in the 176-Kd fragment. However, instead of the loss of larger forms of normal multimers reported by other investigators, the plasma of all but one of our patients (whether they had TTP or HUS) contained a set of larger than normal (supranormal) multimers. Hence, although proteolytic fragmentation of vWF was enhanced during acute TTP/HUS, this phenomenon was not associated with the loss of larger multimers. In the five patients who survived the acute disease and underwent plasma exchange (three with HUS and two with chronic relapsing TTP), subunits and fragments returned to normal values, and supranormal multimers were no longer detected in plasma. In conclusion, even though vWF proteolysis is enhanced in acute TTP/HUS, it does not lead to loss of larger multimers.


Generating segmental mutations in haloalkane dehalogenase: a novel part in the directed evolution toolbox.
Directed evolution techniques allow us to genuinely mimic molecular evolution in vitro. To enhance this imitation of natural evolutionary processes on a laboratory scale in even more detail, we developed an in vitro method for the generation of random deletions and repeats. The pairwise fusion of two fragments of the same gene that are truncated by exonuclease BAL-31 either at the 3' or 5' side results in a deletion or a repeat at the fusion point. Although in principle the method randomly covers the whole gene, it can also be limited to a predefined area in the sequence by controlling the level of the initial truncation. To test the procedure and to illustrate its potential, we used haloalkane dehalogenase from Xanthobacter autotrophicus GJ10 (DhlA) as a model enzyme, since the adaptation of this enzyme towards new substrates is known to occur via the generation of this type of mutation. The results show that the mutagenesis method presented here is an effective tool for accessing formerly unexplorable sequence space and can contribute to the success of future directed evolution experiments.


SEISMOLOGY: Watching the Hayward Fault.
The Hayward fault, located on the east side of the San Francisco Bay, represents a natural laboratory for seismologists, because it does not sleep silently between major earthquakes. In his Perspective, Simpson discusses the study by Bürgmann et al., who have used powerful new techniques to study the fault. The results indicate that major earthquakes cannot originate in the northern part of the fault. However, surface-rupturing earthquakes have occurred in the area, suggesting that they originated to the north or south of the segment studied by Bürgmann et al. Fundamental questions remain regarding the mechanism by which plate tectonic stresses are transferred to the Hayward fault.


Size and behavior in ants: Constraints on complexity.
In this paper I investigate the behavioral complexity of ants in relation to brain size. The volume of the corpora pedunculata, involved in the selection of motor programs, and the antennal lobes, involved in processing olfactory information, are directly related to the volume of the brain of an ant. The volume of the optic lobe, involved in visual processing, is not related to total brain volume. Brain volume is allometrically related to head width in ants. Behavioral complexity, as assayed by the size of the behavioral repertoire within the nest, is directly related to the cube of head width. Behavioral complexity is thus related to the 3/2 power of brain volume. The size of the behavioral repertoire does not appear to correlate with any ecological characteristics of the species but may be a passive consequence of body size. Determination of body size by numerous ecological factors may place constraints on the complexity of behavior that an ant can achieve.