The Cease Smoking Today (CS2day) initiative: a guide to pursue the 2010 IOM report vision for CPD.
This article reviews the articles in this supplement that describe a smoking cessation project, Cease Smoking Today (CS2day) that demonstrated successful outcomes: physician adoption of a smoking cessation guideline and an increase in smoking quit rates. The authors examine how the activities of the CS2day project compared to the principles and characteristics of the vision for a future system of continuing professional development that was described in the 2010 Institute of Medicine (IOM) report, Redesigning Continuing Education in the Health Professions. While it did not meet all the characteristics of the IOM's vision, the CS2day Initiative met enough of them for the authors to suggest that the momentum that the IOM report generated has been sustained. The authors point out two strengths and two weaknesses that further strategic and organizational efforts should consider. The two strengths were the collaborative nature of the project and an approach to continuous outcomes assessment that was based on competencies derived from a practice guideline. The authors also suggested that future similar efforts should place more emphasis on instructional design and developing a program theory to guide program operations and educational development. The authors encouraged members of the CME profession to consider the accomplishments of the CS2day project as they are planning to move their CME programs toward the vision described in the IOM Report.


Interaction of 5-methyltetrahydrofolate and tetrahydrobiopterin on endothelial function.
The present study was designed to investigate the interaction between 5-methyltetrahydrofolate and tetrahydrobiopterin in modulating endothelial function. Tetrahydrobiopterin is a critical cofactor for nitric oxide synthase and maintains this enzyme as a nitric oxide- versus superoxide-producing enzyme. The structure of 5-methyltetrahydrofolate is similar to tetrahydrobiopterin and both agents have been shown to improve endothelium-dependent vasodilatation. We hypothesized that 5-methyltetrahydrofolate interacts with nitric oxide synthase in a fashion analogous, yet independent, of tetrahydrobiopterin to improve endothelial function. We demonstrate that 5-methyltetrahydrofolate binds the active site of nitric oxide synthase and mimics the orientation of tetrahydrobiopterin. Furthermore, 5-methyltetrahydrofolate attenuates superoxide production (induced by inhibition of tetrahydrobiopterin synthesis) and improves endothelial function in aortae isolated from tetrahydrobiopterin-deficient rats. We suggest that 5-methyltetrahydrofolate directly interacts with nitric oxide synthase to promote nitric oxide (vs. superoxide) production and improve endothelial function. 5-Methyltetrahydrofolate may represent an important strategy for intervention aimed at improving tetrahydrobiopterin bioavailability.


Heterologous expression of AtClo1, a plant oil body protein, induces lipid accumulation in yeast.
Proteomic approaches on lipid bodies have led to the identification of proteins associated with this compartment, showing that, rather than the inert fat depot, lipid droplets appear as complex dynamic organelles with roles in metabolism control and cell signaling. We focused our investigations on caleosin [Arabidopsis thaliana caleosin 1 (AtClo1)], a minor protein of the Arabidopsis thaliana seed lipid body. AtClo1 shares an original triblock structure, which confers to the protein the capacity to insert at the lipid body surface. In addition, AtClo1 possesses a calcium-binding domain. The study of plants deficient in caleosin revealed its involvement in storage lipid degradation during seed germination. Using Saccharomyces cerevisiae as a heterologous expression system, we investigated the potential role of AtClo1 in lipid body biogenesis and filling. The green fluorescent protein-tagged protein was correctly targeted to lipid bodies. We observed an increase in the number and size of lipid bodies. Moreover, transformed yeasts accumulated more fatty acids (+46.6%). We confirmed that this excess of fatty acids was due to overaccumulation of lipid body neutral lipids, triacylglycerols and steryl esters. We showed that the original intrinsic properties of AtClo1 protein were sufficient to generate a functional lipid body membrane and to promote overaccumulation of storage lipids in yeast oil bodies.


Femtosecond infrared spectroscopy of chlorophyll f-containing photosystem I.
The recent discovery of extremely red-shifted chlorophyll f pigments in both photosystem I (PSI) and photosystem II has led to the conclusion that chlorophyll f plays a role not only in the energy transfer, but also in the charge separation processes [Nürnberg et al., Science, 2018, 360, 1210-1213]. We have employed ultrafast transient infrared absorption spectroscopy to study the contribution of far-red light absorbing chlorophyll f to energy transfer and charge separation processes in far-red light-grown PSI (FRL-PSI) from the cyanobacterium Chroococcidiopsis thermalis PCC 7203. We compare the kinetics and spectra of FRL-grown PSI excited at 670 nm and 740 nm wavelengths to those of white light-grown PSI (WL-PSI) obtained at 675 nm excitation. We report a fast decay of excited state features of chlorophyll a and complete energy transfer from chlorophyll a to chlorophyll f in FRL-PSI upon 670 nm excitation, as indicated by a frequency shift in a carbonyl absorption band occurring within a 1 ps timescale. While the WL-PSI measurements support the assignment of initial charge separation to A-1+˙A0-˙ [Di Donato et al., Biochemistry, 2011, 50, 480-490] from the kinetics of a distinct cation feature at 1710 cm-1, in the case of FRL-PSI, small features at 1715 cm-1 from the chlorophyll cation are present from sub-ps delays instead, supporting the replacement of the A-1 pigment with chlorophyll f. Comparisons of nanosecond spectra show that charge separation proceeds with 740 nm excitation, which selectively excites chlorophyll f, and modifications in specific carbonyl absorption bands assigned to P700+˙ minus P700 and A1-˙ minus A1 indicate dielectric differences of FRL-PSI compared to WL-PSI in one or both of the two electron transfer branches of FRL-PSI.


Snakebite doesn't cause disseminated intravascular coagulation: coagulopathy and thrombotic microangiopathy in snake envenoming.
The most common coagulopathy associated with snake envenoming worldwide is venom-induced consumption coagulopathy (VICC), which results from activation of the coagulation pathway by snake toxins including thrombin-like enzymes, prothrombin activators, and factor X activators. VICC has often been likened to disseminated intravascular coagulation (DIC) because of the elevated D-dimer, prolonged prothrombin time, and low fibrinogen. However, VICC is not characterized by other important features of DIC, such as evidence of systemic microthrombi and end-organ failure. In addition, the time course of VICC differs with rapid onset and resolution, and the mechanism of initiation of coagulation activation differs because thrombin generation in DIC is mediated by the tissue factor/factor VIIa pathway. In a proportion of patients with VICC, a clinical syndrome consistent with thrombotic microangiopathy has been reported and is characterized by acute renal failure, thrombocytopenia, and microangiopathic hemolytic anemia. This thrombotic microangiopathy appears to only occur in conjunction with VICC but in several different snakes worldwide including vipers and elapids. Consistent with thrombotic microangiopathy, it progresses despite the resolution of the coagulopathy, suggesting a distinct but related process. The existence of the overlapping clinical syndromes of VICC and thrombotic microangiopathy in snake envenoming is the likely reason for the mistaken idea that snakebite causes DIC.


The training characteristics of severely head-injured patients: a preliminary study.
It is argued that an important aspect of rehabilitation after severe head injury lies in training or retraining victims in skills required for independent living. Despite this, very little is known about the training characteristics of head injured patients. It is shown that in the acquisition of a psychomotor skill severely head injured subjects have very poor starting levels as compared to normal controls. They show impressive learning to the extent that they considerably close the gap between themselves and the controls. They also show appreciable transfer of the learned skill from one version of the task to another. The findings are discussed in terms of their implications for further understanding of the training process in head injured subjects in ways that could reflect upon clinical practice in rehabilitation.


Proteomic technologies for the identification of disease biomarkers in serum: advances and challenges ahead.
Serum is an ideal biological sample that contains an archive of information due to the presence of a variety of proteins released by diseased tissue, and serum proteomics has gained considerable interest for the disease biomarker discovery. Easy accessibility and rapid protein changes in response to disease pathogenesis makes serum an attractive sample for clinical research. Despite these advantages, the analysis of serum proteome is very challenging due to the wide dynamic range of proteins, difficulty in finding low-abundance target analytes due to the presence of high-abundance serum proteins, high levels of salts and other interfering compounds, variations among individuals and paucity of reproducibility. Sample preparation introduces pre-analytical variations and poses major challenges to analyze the serum proteome. The label-free detection techniques such as surface plasmon resonance, microcantilever, few nanotechniques and different resonators are rapidly emerging for the analysis of serum proteome and they have exhibited potential to overcome few limitations of the conventional techniques. In this article, we will discuss the current status of serum proteome analysis for the biomarker discovery and address key technological advancements, with a focus on challenges and amenable solutions.


Impact of tuberculosis control measures and crowding on the incidence of tuberculous infection in Maryland prisons.
Our aim was to determine the incidence of tuberculin skin test (TST) conversion in the Maryland state correctional system. We conducted a historical longitudinal cohort study. A sample of 1,289 inmates, incarcerated in 16 of 23 prisons, who had a negative TST and a second test within 24 months was selected. The incidence of recent conversion was 6.3 per 100 person-years. Risk factors for conversion included high prison-population density (relative risk [RR] = 2.4; 95% confidence interval [CI], 1.5-3.8) and incarceration in a higher-security institution (RR = 2.4; 95% CI, 1.4-4.3). Incarceration in an institution with higher levels of isoniazid prophylaxis (> 65% of TST positives) reduced the risk of infection by 50% (RR = 0.5; 95% CI, 0.3-0.7). Crowding was strongly correlated with risk of conversion (r = 0.83; P < .001), while rates of isoniazid prophylaxis initiation were inversely correlated with risk of infection (r = -0.82; P < .001). In stepwise regression, higher prison-population density was the strongest predictor of increased infection. In a final model, inclusion of the rate of isoniazid prophylaxis initiation reduced the risk associated with crowding (RR = 1.4; P = .4). Annual screening programs for prisons can identify recent conversions that may not otherwise be detected.


[Absorption of essential amino acids in patients with chronic enteritis].
Continuous perfusion was applied in studying the absorption of essential amino acids in the duodenum and the proximal part of the jejunum in healthy persons and in patients with chronic enteritis of the land ii degrees of severity. All the patients demonstrated a reduced rate of absorption. The degree of the absorption derangement increased parallel with the severity of the affection. In the duodenum the absorption proceeded slower than in the upper portion of the jejunum.


Regulation of myocardial oxygen delivery in response to graded reductions in hematocrit: role of K+ channels.
This study was designed to identify mechanisms responsible for coronary vasodilation in response to progressive decreases in hematocrit. Isovolemic hemodilution was produced in open-chest, anesthetized swine via concurrent removal of 500 ml of arterial blood and the addition of 500 ml of 37 °C saline or synthetic plasma expander (Hespan, 6% hetastarch in 0.9% sodium chloride). Progressive hemodilution with Hespan resulted in an increase in coronary flow from 0.39 ± 0.05 to 1.63 ± 0.16 ml/min/g (P < 0.001) as hematocrit was reduced from 32 ± 1 to 10 ± 1% (P < 0.001). Overall, coronary flow corresponded with the level of myocardial oxygen consumption, was dependent on arterial pressures ≥ ~ 60 mmHg, and occurred with little/no change in coronary venous PO2. Anemic coronary vasodilation was unaffected by the inhibition of nitric oxide synthase (L-NAME: 25 mg/kg iv; P = 0.92) or voltage-dependent K+ (K V) channels (4-aminopyridine: 0.3 mg/kg iv; P = 0.52). However, administration of the K ATP channel antagonist (glibenclamide: 3.6 mg/kg iv) resulted in an ~ 40% decrease in coronary blood flow (P < 0.001) as hematocrit was reduced to ~ 10%. These reductions in coronary blood flow corresponded with significant reductions in myocardial oxygen delivery at baseline and throughout isovolemic anemia (P < 0.001). These data indicate that vasodilator factors produced in response to isovolemic hemodilution converge on vascular smooth muscle glibenclamide-sensitive (K ATP) channels to maintain myocardial oxygen delivery and that this response is not dependent on endothelial-derived nitric oxide production or pathways that mediate dilation via K V channels.


Death from minor head trauma and alcohol.
Sudden death in association with minor or otherwise insignificant head injury is increasingly being recognised in the forensic medical literature. While the exact mechanism of the cardiorespiratory arrest that appears to occur in these cases is unclear, a number of mechanisms have been postulated. Animal studies have provided evidence that alcohol can be associated with an increased period of apnoea following minor physical brain injury and cardiac changes have also been identified. The limited information in the community about the risk of alcohol in association with minor head injury causing death may be relevant in homicide cases where this issue is raised.


A nationwide survey of antibiotic prescribing patterns and clinical outcomes in patients with bacterial pneumonia.
Antibiotic prescribing information was prospectively collected on 1822 hospitalized patients treated for suspected or documented bacterial pneumonia. Antibacterial therapy with a single antibiotic was employed in more than 50 percent of the patients, with cefazolin, cefuroxime, ampicillin, and ceftriaxone sodium representing the most commonly employed agents. Combination therapy using two antibiotics was employed in approximately 30 percent of patients with the aminoglycosides, particularly gentamicin, used extensively. A satisfactory outcome was achieved in approximately 80 percent of patients with a community- or institutional-acquired pneumonia; only 66 percent of nosocomial pneumonias had a satisfactory outcome. An important observation was the apparently common practice of switching patients to an oral antibiotic regimen after an average of seven days of antibiotics and subsequently discharging the patient. No difference was observed in the patterns of clinical response or duration of therapy for culture-positive versus culture-negative patients. The results of this surveillance program can serve as a basis for comparison of institution-specific drug utilization evaluation programs.


An evolving redefinition of autoimmune encephalitis.
Autoimmune encephalitis encompasses a wide variety of protean pathologic processes associated with the presence of antibodies against neuronal intracellular proteins, synaptic receptors, ion channels and/or neuronal surface proteins. This type of encephalitis can also involve children with complex patterns of seizures and unexpected behavioural changes, which jeopardize their prompt recognition and treatment. Many epidemiological studies have shown that numerous immune-based forms of encephalitis can be encountered, almost surpassing the rate of postinfectious encephalitides. However, the overall exact prevalence of autoimmune encephalopathies remains underestimated, and the definition of diagnostic algorithms results muddled. The spectrum of neuropsychiatric manifestations in the pediatric population with autoimmune encephalitis is less clear than in adults, but the integration of clinical, immunological, electrophysiological and neuroradiological data is essential for a general approach to patients. In this review we report the most relevant data about both immunologic and clinical characteristics of the main autoimmune encephalitides recognized so far, with the aim of assisting clinicians in the differential diagnosis and favouring an early effective treatment. Correlations between phenotype and autoantibodies involved in the neurological damage of autoimmune encephalitis are largely unknown in the first years of life, because of the relatively small number of pediatric patients adequately studied. Future multicenter collaborative studies are needed to improve the diagnostic approach and tailor personalized therapies in the long-term.


The role of surgery in traumatic brain injury.
To identify the surgical indications in the treatment of posttraumatic intracranial hematomas and to evaluate the role of external decompression in severe posttraumatic intracranial hypertension. An evidence-based review resulted recently in the publication of Guidelines for Surgical Management of Traumatic Brain Injury. Unfortunately all published surgical studies are at the level of simple options with no standards. We know that a number of patients harbouring small epidural/subdural hematomas can be managed conservatively when the lesions are less than 10 mm of thickness and with a midline shift of less than 5 mm. With exception of these few cases, the current practice is that a large number of patients with posttraumatic hematomas are operated either within 24 h from injury (with acute subdural hematomas as the prevailing lesion) or later (with parenchymal hematomas as the prevailing lesion). According to a recent European study, about one third of these patients are also decompressed. A surgical approach is frequent in posttraumatic intracranial hematomas in spite of a low level of evidence. One of the surgical options either in association with hematoma evacuation or in isolation is the technique of bone flap decompression.


Effect of influenza hemagglutinin fusion peptide on lamellar/inverted phase transitions in dipalmitoleoylphosphatidylethanolamine: implications for membrane fusion mechanisms.
Low mole fractions of viral fusion peptides induce inverted cubic (Q(II)) phases in dipalmitoleoylphosphatidylethanolamine (DiPoPE), a lipid with unsaturated acyl chains that normally forms inverted hexagonal phase (H(II)) above 43 degrees C. The ability to form a Q(II) phase is relevant to the study of membrane fusion: fusion occurs in liposomal systems under conditions where Q(II) phase precursors form, and fusion may be an obligatory step in the lamellar (L(alpha))/Q(II) phase transition. We used X-ray diffraction and time-resolved cryoelectron microscopy (TRC-TEM) to study the effects of the influenza hemagglutinin fusion peptide on the phase behavior and structure of DiPoPE. X-ray diffraction data show that at concentrations of 3-7 mol%, the fusion peptide (FP) induces formation of a Q(II) phase in preference to the H(II) phase. TRC-TEM data show that the FP acts at early stages in the phase transition (i.e. within seconds): at 2-7 mol%, FP decreases or inhibits formation of the L(alpha)/H(II) intermediate morphology observed via TRC-TEM in pure DiPoPE at the same temperature. Our X-ray diffraction data imply that FP either does not affect, or slightly increases, the spontaneous curvature of the host lipid (i.e. either does not affect or tends to destabilize inverted phases, respectively). FP may act in part by affecting the relative stability of two intermediate structures in the phase transition mechanism, as suggested previously. These results indicate a new way in which hydrophobic sequences of membrane proteins may be fusogenic.


Evidence-based kernels: fundamental units of behavioral influence.
This paper describes evidence-based kernels, fundamental units of behavioral influence that appear to underlie effective prevention and treatment for children, adults, and families. A kernel is a behavior-influence procedure shown through experimental analysis to affect a specific behavior and that is indivisible in the sense that removing any of its components would render it inert. Existing evidence shows that a variety of kernels can influence behavior in context, and some evidence suggests that frequent use or sufficient use of some kernels may produce longer lasting behavioral shifts. The analysis of kernels could contribute to an empirically based theory of behavioral influence, augment existing prevention or treatment efforts, facilitate the dissemination of effective prevention and treatment practices, clarify the active ingredients in existing interventions, and contribute to efficiently developing interventions that are more effective. Kernels involve one or more of the following mechanisms of behavior influence: reinforcement, altering antecedents, changing verbal relational responding, or changing physiological states directly. The paper describes 52 of these kernels, and details practical, theoretical, and research implications, including calling for a national database of kernels that influence human behavior.


Oviductal antibody response to a defined recombinant sperm antigen in macques.
Macaque oviductal fluids were assayed for specific antibodies to the intra-acrosomal sperm protein SP-10 after immunizations with recombinant macaque SP-10 (re-mqSP-10), a candidate contraceptive vaccinogen. Access ports, consisting of a subcutaneous collecting reservoir and a catheter to cannulate the oviduct, were implanted into monkeys for repeated aspiration of oviductal fluid. Monkeys were inoculated i.m. once a month with an emulsion consisting of 2 mg re-mqSP-10 in a vehicle of squalene and mannin monooleate. Oviductal fluids and serum were collected during the periovulatory period for six menstrual cycles, and IgG and IgA antigen-specific antibodies in preimmune and immune fluids were compared by ELISA. Both relative and absolute concentrations of SP-10-specific immunoglobulins (Ig) were determined. Oviductal fluids from immunized animals showed significant increases in anti-SP-10 IgG at cycle 2 and at all subsequent intervals. Anti-SP-10 IgA significantly increased in oviductal fluid at cycles 4, 5, and 6. Serum anti-SP-10 IgG increased at cycle 2 and remained significantly elevated through cycle 6, while serum anti-SP-10 IgA was higher than in preimmune samples at cycle 4. Serum antibodies generated to the recombinant SP-10 recognized SP-10 extracted from macaque sperm on Western blots. Immunocytochemical staining of macaque and human sperm showed acrosomal immunofluorescence with both immune oviductal fluids and serum using both anti-IgG and anti-IgA secondary antibodies. This study demonstrates for the first time 1) IgG and IgA antibodies to a defined recombinant sperm-specific antigen in primate oviductal fluids after systemic immunization and 2) the recognition by primate oviductal fluid IgG and IgA of the endogenous contraceptive target on both human and macaque sperm.


Arabinan-deficient mutants of Corynebacterium glutamicum and the consequent flux in decaprenylmonophosphoryl-D-arabinose metabolism.
The arabinogalactan (AG) of Corynebacterianeae is a critical macromolecule that tethers mycolic acids to peptidoglycan, thus forming a highly impermeable cell wall matrix termed the mycolyl-arabinogalactan peptidoglycan complex (mAGP). The front line anti-tuberculosis drug, ethambutol (Emb), targets the Mycobacterium tuberculosis and Corynebacterium glutamicum arabinofuranosyltransferase Mt-EmbA, Mt-EmbB and Cg-Emb enzymes, respectively, which are responsible for the biosynthesis of the arabinan domain of AG. The substrate utilized by these important glycosyltransferases, decaprenylmonophosphoryl-D-arabinose (DPA), is synthesized via a decaprenylphosphoryl-5-phosphoribose (DPPR) synthase (UbiA), which catalyzes the transfer of 5-phospho-ribofuranose-pyrophosphate (pRpp) to decaprenol phosphate to form DPPR. Glycosyl compositional analysis of cell walls extracted from a C. glutamicum::ubiA mutant revealed a galactan core consisting of alternating beta(1-->5)-Galf and beta(1-->6)-Galf residues, completely devoid of arabinan and a concomitant loss of cell-wall-bound mycolic acids. In addition, in vitro assays demonstrated a complete loss of arabinofuranosyltransferase activity and DPA biosynthesis in the C. glutamicum::ubiA mutant when supplemented with p[14C]Rpp, the precursor of DPA. Interestingly, in vitro arabinofuranosyltransferase activity was restored in the C. glutamicum::ubiA mutant when supplemented with exogenous DP[14C]A substrate, and C. glutamicum strains deficient in ubiA, emb, and aftA all exhibited different levels of DPA biosynthesis.


Relationship between trust and patient involvement in medical decision-making: A cross-sectional study.
Patients vary in their preferences regarding involvement in medical decision-making. Current research does not provide complete explanation for this observed variation. Patient involvement in medical decision-making has been found to be influenced by various mechanisms, one of which could be patients' trust in physicians. The aim of this study was to examine whether trust in physicians fosters or impairs patient involvement in medical decision-making. This study also aimed to determine to what extent the relationship between trust and preferences regarding decision-making roles was influenced by the sociodemographic characteristics of the patients. We hypothesised that trust can both foster and impair patient involvement in medical decision-making. A survey was sent out to members of the Nivel Dutch Health Care Consumer Panel in February 2016 (response rate = 47%, N = 703). The Wake Forest Physician Trust Scale was used to measure trust. Patient involvement was measured using two items based on the study published by Flynn and colleagues in 2006. Multiple regression analysis was used to analyse the relationship between trust and patient involvement. We found a negative relationship between trust and patient involvement in medical decision-making in men. Women with high trust reported to be more involved in medical decision-making compared to men with high trust. The results suggest that trust impairs involvement in medical decision-making for men but not for women. Further research could provide a more comprehensive explanation of the variation in patient preferences regarding involvement in medical decision-making to further elucidate which underlying mechanisms could enhance patient participation.


High-resolution optical coherence microscopy for high-speed, in vivo cellular imaging.
Optical coherence microscopy (OCM) is demonstrated with a high-speed, broadband, reflective-grating phase modulator and a femtosecond Ti:Al2O3 laser. The novel system design permits high-resolution OCM imaging in a new operating regime in which a short coherence gate is used to relax the requirement for high-numerical-aperture confocal axial sectioning. In vivo cellular imaging is demonstrated in the Xenopus laevis tadpole and in human skin with a 3-microm coherence gate and a 30-microm confocal gate. The ability to achieve cellular imaging with a lower numerical aperture should facilitate the development of miniaturized probes for in vivo imaging applications.


A higher resolution radiation hybrid map of bovine chromosome 13.
In this paper, we present a radiation hybrid framework map of BTA13 composed of nine microsatellite loci, six genes and one EST. The map has been developed using a recently constructed 12'000 rad bovine-hamster whole-genome radiation hybrid panel. Moreover, we present a comprehensive map of BTA13 comprising 72 loci, of which 45 are microsatellites, 20 are genes and seven are ESTs. The map has an estimated length of 2694.7 cR(12'000). The proposed order is in general agreement with published maps of BTA13. Our results only partially support previously published information of five blocks of conserved gene order between cattle and man. We found no evidence for the existence of an HSA20 homologous segment of coding DNA on BTA13 located centromeric of a confirmed HSA10 homologous region. The present map increases the marker density and the marker resolution on BTA13 and enables further insight into the evolutionary development of the chromosome as compared to man.


Target resolution in visual search involves the direct suppression of distractors: evidence from electrophysiology.
Attentional selection requires both the enhancement of target stimuli and the suppression of distractors. Target enhancement and distractor suppression have been associated with separable components of the lateralized event-related potential: the target negativity and distractor positivity (Pd). We examined the distractor suppression mechanisms indexed by the Pd. The Pd may reflect mechanisms that guide attention away from distractors during search or mechanisms involved in the subsequent resolution of target features. To determine which of these alternatives was more likely, we had participants view search arrays that contained only a target, only a distractor, or both. The Pd elicited by distractors was substantially larger when the display also contained a target, consistent with the idea that this component reflects a mechanism of distractor suppression activated during the resolution and disambiguation of target features.


Physical factors in cataractogenesis: ambient ultraviolet radiation and temperature.
A number of environmental cofactors have been implicated in cataracto-genesis. Two have received the greatest attention: ultraviolet radiation (UVR) and ambient temperature. Unfortunately, both temperature and UVR levels vary similarly with geographical latitude. Careful attention to several more refined physical variables and the geometry of exposure may permit investigators to separate the contributory effects of these two physical agents. This paper briefly reviews the available data, estimates the variation of lenticular temperature with ambient temperature, and provides measurements of short-wavelength (UV-B) UVR exposure to the human eye with different meterological conditions. The study attempts to provide epidemiological investigators with more detailed information necessary to perform more accurate studies of cataract and other ocular pathologies that appear to be related to environmental factors. Ocular UV-B radiation exposure levels were measured at nine locations in the USA near 40 degrees latitude at elevations from sea level to 8000 ft. Terrain reflectance is shown to be much more important than terrain elevation; cloud cover and haze may actually increase ocular exposure; and the value of wearing brimmed hats and spectacles varies with the environment. Several avenues for future research are suggested.


Reduced pericardial levels of endostatin correlate with collateral development in patients with ischemic heart disease.
We investigated whether pericardial levels of a pro-angiogenic factor (vascular endothelial growth factor, VEGF) or an anti-angiogenic factor (endostatin) related to the presence of coronary collateral circulation in patients with significant coronary artery disease (CAD). Coronary collateralization favorably alters the prognosis of patients with occlusive CAD. The specific factors that mediate and maintain collateral formation in coronary vessel occlusion are yet to be identified. Coronary angiograms from 39 patients undergoing coronary artery bypass surgery were evaluated for the absence of collaterals (n = 20) or the presence of Rentrop classification grade 3 collaterals (n = 19). Pericardial fluid samples were obtained at the time of surgery and were assayed for the VEGF and endostatin by enzyme-linked immunosorbent assay comparing the two groups of patients. Vascular endothelial growth factor levels were not significantly different between the groups (28.86 +/- 4.67 pg/ml vs. 24.39 +/- 3.08 pg/ml, p = 0.43). However, pericardial fluid endostatin levels were nearly 40% lower in patients with grade 3 collateralization compared with those lacking angiographic evidence of collaterals (15.17 +/- 1.87 ng/ml vs. 24.25 +/- 2.08 ng/ml, p < 0.0025). Pericardial fluid levels of endostatin, but not VEGF, are associated with the presence or absence of collaterals in patients with CAD. These data suggest that the angiogenesis inhibitor endostatin levels may locally modulate coronary collateral formation.


Formation of superoxide anion in the hepatic sinusoid after lipopolysaccharide challenge.
Using the cytochrome c method, superoxide anion that is released into the hepatic sinusoid was measured after a lipopolysaccharide challenge in a liver perfusion system. Moreover, damages of epithelial cells of the hepatic sinusoid were estimated with scanning electron microscopic analysis and levels of purine nucleoside phosphorylase/GPT ratio. Lipopolysaccharide administration increased the conversion of oxidized cytochrome c into reduced cytochrome c in the perfusate, indicating that superoxide anion was formed in the hepatic sinusoid. This change was associated with increase in levels of portal tumor necrosis factor-alpha and attenuated by the simultaneous administration of superoxide dismutase. Scanning electron microscope analysis revealed that diameters of sinusoidal fenestrae increased in rats treated with lipopolysaccharide, compared with controls. Moreover, levels of purine nucleoside phosphorylase/GPT ratio was significantly increased in the liver perfusate in lipopolysaccharide-treated rats, compared with controls. Superoxide anion in hepatic sinusoid may be one of the pathogenic factors behind damages of epithelial cells of the hepatic sinusoid caused by lipopolysaccharide.


Pancreaticoduodenectomy for periampullary carcinoma.
To assess the morbidity, mortality and 1- and 2-year survival rates, and safety of pancreaticoduodenectomy for periampullary (including pancreatic head) carcinomas in a non-oncology surgical set-up. Records of 45 patients undergoing pancreaticoduodenectomies for periampullary cancers between July 1996 and April 2000 were reviewed. These included ampullary (n=23), pancreatic (14) and duodenal (2) adenocarcinomas, lower-end cholangiocarcinoma (5), and ampullary carcinoid (1). Thirty-seven patients underwent the Whipple procedure and 8 underwent the pylorus-preserving modification. The overall mortality rate was 11% and morbidity rate was 46%. Wound infection was the most common postoperative complication. The 1- and 2-year survival rates for periampullary cancers were 61% and 39% and those for pancreatic cancers were 57% and 36%, respectively. Pancreaticoduodenectomy for periampullary tumors remains a formidable procedure in our set-up. However, it can be performed safely with low mortality and morbidity rates.


High-Speed Automated Reconstruction of Drosophila Larval Brain from Volumetric EM Data.
To uncover the relationship between neural activity and behavior, it is essential to reconstruct neural circuits. However, methods typically used for neuron reconstruction from volumetric electron microscopy (EM) dataset are often time-consuming and require extensive manual proofreading, making it difficult to reproduce in a typical laboratory setting. To address this challenge, we have developed a set of acceleration techniques that build upon the Flood Filling Network (FFN), significantly reducing the time required for this task. These techniques can be easily adapted to other similar datasets and laboratory settings. To validate our approach, we tested our pipeline on a dataset of Drosophila larval brain serial section EM images at synaptic-resolution level. Our results demonstrate that our pipeline significantly reduces the inference time compared to the FFN baseline method and greatly reduces the time required for reconstructing the 3D morphology of neurons.


Evaluation of dietary selenium, vitamin C and E as the multi-antioxidants on the methylmercury intoxicated mice based on mercury bioaccumulation, antioxidant enzyme activity, lipid peroxidation and mitochondrial oxidative stress.
Mercury (Hg) in high exposures can be a potent life threatening heavy metal that bioaccumulate in aquatic food-chain mainly as organic methylmercury (MeHg). In this regard, fish and seafood consumptions could be the primary sources of MeHg exposure for human and fish-eating animals. The objective of the present study was to elucidate the effects of dietary supplementation of some antioxidants on induced mercury toxicity in mice model. In this study, a 30-day long investigation has been conducted to evaluate the dietary effect of selenium (Se) in combination with vitamin C and vitamin E on methylmercury induced toxicity in mice. Total 54 mice fed the diets with three levels of Hg (0, 50 or 500 μg kg-1) and two levels of Se in combination with vitamin C and E (Se: 0, 2 mg kg-1; vitamin C: 0, 400 mg kg-1; vitamin E: 0, 200 mg kg-1) in triplicates. The results show that Hg accumulated in blood and different tissues such as muscle, liver and kidney tissues of mice on dose dependent manner. The bioaccumulation pattern of dietary Hg, in decreasing order, kidney > liver > muscle > blood. Superoxide dismutase levels in blood serum showed no significant differences in mice fed the diets. However, dietary antioxidants significantly reduced the levels of thiobarbituric acid reactive substances in mice fed the mercury containing diets. Cytochrome c oxidase enzyme activities showed no significant differences as the mercury level increases in liver and kidney tissues of mice. Kaplan-Meier curve showed a dose- and time-dependent survivability of mice. Cumulative survival rate of Hg intoxicated mice fed the antioxidant supplemented diets were increased during the experimental period. Overall, the results showed that dietary Se, vitamin C and vitamin E had no effect on reducing the mercury bioaccumulation in tissues but reduced the serum lipid peroxidation as well as prolonged the cumulative survival rate in terms of high Hg exposures in mice.


Amorphous CeOx Islands on Dealuminated Zeolite Beta to Stabilize Pt Nanoparticles as Efficient and Antisintering Catalysts for Propane Dehydrogenation.
Pt-based catalysts have been widely used in propane dehydrogenation due to their superior activation of C-H bonds and weak scission of C-C bonds. However, in the process of repeated calcination to remove deposited coke, the active Pt species tend to sinter, resulting in a significant decline in catalytic activity. In this study, amorphous CeOx islands loaded on dealuminated Beta zeolite were prepared via simple wetness impregnation. Then, partially embedded Pt nanoparticles in CeOx islands were obtained after reduction owing to the affinity of CeOx for Pt. In the propane dehydrogenation reaction, Pt/Ce5-SiBeta with a Ce loading of 4.55 wt % and Pt loading of 0.72 wt % exhibited the highest activity and the lowest inactivation constant at 550 °C. More importantly, due to the anchoring effect of CeOx on Pt, the catalytic activity of Pt could be recovered after a simple calcination-reduction regeneration process, avoiding the chlorination treatment for the redispersion of Pt species used in industry. In addition, to improve the selectivity of the Pt/Ce5-SiBeta catalyst, a PtSn/Ce5-SiBeta catalyst with excellent activity, selectivity, and recycling stability has been prepared by introducing Sn into Pt/Ce5-SiBeta. The use of amorphous CeOx islands to improve the sintering resistance of Pt opens up new prospects for the design of stable industrial dehydrogenation catalysts.


Immunomodulatory properties of CD38 antibodies and their effect on anticancer efficacy in multiple myeloma.
CD38 has been established as an important therapeutic target for multiple myeloma (MM), for which two CD38 antibodies are currently approved-daratumumab and isatuximab. CD38 is an ectoenzyme that degrades NAD and its precursors and is involved in the production of adenosine and other metabolites. Among the various mechanisms by which CD38 antibodies can induce MM cell death is immunomodulation, including multiple pathways for CD38-mediated T-cell activation. Patients who respond to anti-CD38 targeting treatment experience more marked changes in T-cell expansion, activity, and clonality than nonresponders. Resistance mechanisms that undermine the immunomodulatory effects of CD38-targeting therapies can be tumor intrinsic, such as the downregulation of CD38 surface expression and expression of complement inhibitor proteins, and immune microenvironment-related, such as changes to the natural killer (NK) cell numbers and function in the bone marrow niche. There are numerous strategies to overcome this resistance, which include identifying and targeting other therapeutic targets involved in, for example, adenosine production, the activation of NK cells or monocytes through immunomodulatory drugs and their combination with elotuzumab, or with bispecific T-cell engagers.


Weight loss improves disease activity in patients with psoriatic arthritis and obesity: an interventional study.
Obesity is over-represented in patients with psoriatic arthritis (PsA) and associated with higher disease activity, poorer effect of treatment and increased cardiovascular morbidity. Studies on the effects of weight loss are however needed. This study aimed to prospectively study the effects of weight loss treatment with very low energy diet (VLED) on disease activity in patients with PsA (CASPAR criteria) and obesity (body mass index BMI ≥ 33 kg/m2). VLED (640 kcal/day) was taken during 12-16 weeks, depending on pre-treatment BMI. Afterwards, an energy-restricted diet was gradually reintroduced. Weight loss treatment was given within a structured framework for support and medical follow-up. Treatment with conventional synthetic and/or biologic disease-modifying anti-rheumatic drugs was held constant from 3 months before, until 6 months after baseline. Patients were assessed with BMI, 66/68 joints count, Leeds enthesitis index, psoriasis body surface area (BSA), questionnaires and CRP at baseline, 3 and 6 months. Primary outcome was the percentage of patients reaching minimal disease activity (MDA) and secondary outcomes were reaching Psoriatic Arthritis Response Criteria (PsARC) and American College of Rheumatology (ACR) response criteria. Totally 41/46 patients completed the study, 63% women, median age 54 years (IQR 48-62). At baseline increased BMI was associated with higher disease activity and poorer function. The median weight loss was 18.7 kg (IQR 14.6-26.5) or 18.6% (IQR 14.7-26.3) of the baseline weight. A majority of the disease activity parameters improved significantly after weight loss, including 68/66 tender/swollen joints count, CRP, BSA, Leeds enthesitis index, HAQ and patient VAS for global health, pain and fatigue. A larger weight loss resulted in more improvement in a dose-response manner. The percentage of patients with MDA increased from 29 to 54%, (p = 0.002). PsARC was reached by 46.3%. The ACR 20, 50 and 70 responses were 51.2%, 34.1% and 7.3% respectively. Short-term weight loss treatment with VLED was associated with significant positive effects on disease activity in joints, entheses and skin in patients with PsA and obesity. The study supports the hypothesis of obesity as a promotor of disease activity in PsA. ClinicalTrials.gov identifier: NCT02917434 , registered on September 21, 2016-retrospectively registered.


Bacterial origin and community composition in the barley phytosphere as a function of habitat and presowing conditions.
An understanding of the factors influencing colonization of the rhizosphere is essential for improved establishment of biocontrol agents. The aim of this study was to determine the origin and composition of bacterial communities in the developing barley (Hordeum vulgare) phytosphere, using denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes amplified from extracted DNA. Discrete community compositions were identified in the endorhizosphere, rhizoplane, and rhizosphere soil of plants grown in an agricultural soil for up to 36 days. Cluster analysis revealed that DGGE profiles of the rhizoplane more closely resembled those in the soil than the profiles found in the root tissue or on the seed, suggesting that rhizoplane bacteria primarily originated from the surrounding soil. No change in bacterial community composition was observed in relation to plant age. Pregermination of the seeds for up to 6 days improved the survival of seed-associated bacteria on roots grown in soil, but only in the upper, nongrowing part of the rhizoplane. The potential occurrence of skewed PCR amplification was examined, and only minor cases of PCR bias for mixtures of two different DNA samples were observed, even when one of the samples contained plant DNA. The results demonstrate the application of culture-independent, molecular techniques in assessment of rhizosphere bacterial populations and the importance of the indigenous soil population in colonization of the rhizosphere.


Nutritional performance, hepatic and renal function in goats fed diets containing detoxified castor cake at different stages of pregnancy.
We evaluated the influence of the substitution of soybean meal by detoxified castor cake on performance, digestibility of nutrients, nitrogen balance, hepatic and renal functions of pregnant goats fed with diets containing detoxified castor cake by alkaline solutions during the stages (first two-thirds and final third) of pregnancy. Three diets were provided: one based on soybean meal and the other two based on castor cake detoxified with whit calcium hydroxide or sodium hydroxide. Goats fed detoxified castor cake sodium hydroxide had lower consumption. Was no effect (P>0.05) of diets or stages on the digestibility of dry matter and nutrients. The goats that received the diets based on soybean meal and detoxified castor cake calcium hydroxide consumed larger amounts of nitrogen. The goats fed with diet the basis of SM had greater weight in the parturition day. The average levels of enzymes for hepatic and renal functions were within normal patterns. Of enzymes related to liver metabolism, only the gamma-glutamyl transferase increased in the final third of pregnancy. The present study demonstrated that detoxified castor by sodium hydroxide reduces the consumption of goats during gestation, but did not affect negatively the renal and hepatic parameters.


Unravelling the intrinsic functional organization of the human lateral frontal cortex: a parcellation scheme based on resting state fMRI.
Human and nonhuman primates exhibit flexible behavior. Functional, anatomical, and lesion studies indicate that the lateral frontal cortex (LFC) plays a pivotal role in such behavior. LFC consists of distinct subregions exhibiting distinct connectivity patterns that possibly relate to functional specializations. Inference about the border of each subregion in the human brain is performed with the aid of macroscopic landmarks and/or cytoarchitectonic parcellations extrapolated in a stereotaxic system. However, the high interindividual variability, the limited availability of cytoarchitectonic probabilistic maps, and the absence of robust functional localizers render the in vivo delineation and examination of the LFC subregions challenging. In this study, we use resting state fMRI for the in vivo parcellation of the human LFC on a subjectwise and data-driven manner. This approach succeeds in uncovering neuroanatomically realistic subregions, with potential anatomical substrates including BA 46, 44, 45, 9 and related (sub)divisions. Ventral LFC subregions exhibit different functional connectivity (FC), which can account for different contributions in the language domain, while more dorsal adjacent subregions mark a transition to visuospatial/sensorimotor networks. Dorsal LFC subregions participate in known large-scale networks obeying an external/internal information processing dichotomy. Furthermore, we traced "families" of LFC subregions organized along the dorsal-ventral and anterior-posterior axis with distinct functional networks also encompassing specialized cingulate divisions. Similarities with the connectivity of macaque candidate homologs were observed, such as the premotor affiliation of presumed BA 46. The current findings partially support dominant LFC models.


Homotopy invariance of eta-invariants.
Intersection homology and results related to the higher signature problem are applied to show that certain combinations of eta-invariants of the signature operator are homotopy invariant in various circumstances.


Myeloperoxidase enhances nitric oxide catabolism during myocardial ischemia and reperfusion.
Impaired microvascular function during myocardial ischemia and reperfusion is associated with recruitment of polymorphonuclear neutrophils (PMN) and has been attributed to decreased bioavailability of nitric oxide (NO). Whereas myeloperoxidase (MPO), a highly abundant, PMN-derived heme protein facilitates oxidative NO consumption and impairs vascular function in animal models of acute inflammation, its capacity to function in this regard during human myocardial ischemia and reperfusion remains unknown. Plasma samples from 30 consecutive patients (61 +/- 14 years, 80% male) presenting with acute myocardial infarction were collected 9 +/- 4 h after vessel recanalization and compared to plasma from healthy control subjects (n = 12). Plasma levels of MPO were higher in patients than in control subjects (1.4 +/- 0.9 vs 0.3 +/- 0.2 ng/mg protein, respectively, p < 0.0001). The addition of hydrogen peroxide to patient plasma resulted in accelerated rates of NO consumption compared to control subjects (0.53 +/- 0.25 vs 0.068 +/- 0.039 nM/s/mg protein, respectively, p < 0.0001). Myocardial tissue from patients with the same pathology revealed intense recruitment of MPO-positive PMN localized along infarct-related vessels as well as diffuse endothelial distribution of non-PMN-associated MPO immunoreactivity. Endothelium-dependent microvascular function, as assessed by an acetylcholine-dependent increase in forearm blood flow in 75 patients with symptomatic coronary artery disease, inversely correlated with MPO plasma levels (r = -0.75, p < 0.005). Plasma from patients undergoing myocardial reperfusion contained increased levels of MPO, which catalytically consumed NO in the presence of H(2)O(2). Given the correlation between intravascular MPO levels and forearm vasomotor function in patients with coronary artery disease, MPO appears to be an important modulator of vasomotor function in inflammatory vascular disease and a potential therapeutic target for treatment.


Pediatric ganglioglioma of the brainstem and cervicomedullary junction: a retrospective cohort study.
Ganglioglioma is a low-grade central nervous system neoplasm with a pediatric predominance, accounting for 10% of all brain tumors in children. Gangliogliomas of the cervicomedullary junction (GGCMJs) and brainstem (GGBSs) present a host of management challenges, including a significant risk of surgical morbidity. At present, understanding of the prognostic factors-including BRAF V600E status-is incomplete. Here, the authors report a single-institution GGCMJ and GGBS experience and review the pertinent literature. A prospectively maintained neurosurgical database at a large tertiary care academic referral center was retrospectively queried for cases of GGCMJ pathologically confirmed in the period from 1995 to 2015; appropriate cases were defined by diagnosis codes and keywords. Secondary supplemental chart review was conducted to confirm or capture relevant data. The primary study outcome was treatment failure as defined by evidence of radiographic recurrence or progression and/or clinical or functional decline. A review of the literature was conducted as well. Five neurosurgically managed GGBS patients were identified, and the neoplasms in 4 were classified as GGCMJ. All 5 patients were younger than 18 years old (median 15 years, range 4-16 years) and 3 (60%) were female. One patient underwent gross-total resection, 2 underwent aggressive subtotal resection (STR), and 2 underwent stereotactic biopsy only. All patients who had undergone STR or biopsy required repeat resection for tumor control or progression. Progressive disease was treated with radiotherapy in 2 patients, chemotherapy in 2, and chemoradiotherapy alone in 1. Immunostaining for BRAF V600E was positive in 3 patients (60%). All 5 patients experienced at least one major complication, including wound infection, foot drop, hemiparesis, quadriparesis, cranial neuropathy, C2-3 subluxation, syringomyelia, hydrocephalus, aspiration, and coma. Overall mortality was 20%, with 1 death observed over 11 years of follow-up. GGBS and GGCMJ are rare, benign posterior fossa tumors that carry significant perioperative morbidity. Contemporary management strategies are heterogeneous and include combinations of resection, radiotherapy, and chemotherapy. The BRAF V600E mutation is frequently observed in GGBS and GGCMJ and appears to have both prognostic and therapeutic significance with targeted biological agents.


Response of preosteoblasts on micromachined Ti-6Al-4V surface to microstructure dimension.
The cell incubation depends on the cultured surface, but various machining methods produce different surface topographies, but it has not been clear how it is related to the topographic feature until now. Hence, the machined Ti-6Al-4V surface is characterized for preosteoblasts incubation via different mechanical fabrication. The relationship between surface topography created by various machining methods and cell incubation behaviour was explored. The objective is to control the surface preosteoblasts growth in machining of biological titanium alloy. According to the cell growth kinetic, the cell incubation behaviour was first proposed and modelled in relation to microstructural dimension and culture duration. Then, the topological cultured microstructure surface was fabricated via mechanical fabrication. Finally, the cell initial adhesion and incubation behaviour on microstructured surface was investigated. It is shown that the surface undulation on machined microstructure is conducive to controlling the direction and distribution of cell incubation from cell growth kinetic model. The cell culture can be controlled on the peak with a small undulation, while it is concentred on the sidewall with a high aspect ratio. Increasing the aspect ratio extends cell growth, while low aspect ratio promotes initial cell adhesion and growth rate. Within the optimal cultured duration, the microstructured surface is more favourable for cell survival, and the cell growth keep positive beyond critical aspect ratio. As a result, the cell adhesion ability is topologically controlled to 5.4 times higher and the growth rate can be improved by 101.7% on milled microgrooved surface. It may be applied to the rapid production of biomedical Ti-6Al-4V implant.


Preparation of arginine-glycine-aspartic acid-modified biopolymeric nanoparticles containing epigalloccatechin-3-gallate for targeting vascular endothelial cells to inhibit corneal neovascularization.
Neovascularization (NV) of the cornea can disrupt visual function, causing ocular diseases, including blindness. Therefore, treatment of corneal NV has a high public health impact. Epigalloccatechin-3-gallate (EGCG), presenting antiangiogenesis effects, was chosen as an inhibitor to treat human vascular endothelial cells for corneal NV treatment. An arginine-glycine-aspartic acid (RGD) peptide-hyaluronic acid (HA)-conjugated complex coating on the gelatin/EGCG self-assembly nanoparticles (GEH-RGD NPs) was synthesized for targeting the αvβ3 integrin on human umbilical vein endothelial cells (HUVECs) in this study, and a corneal NV mouse model was used to evaluate the therapeutic effect of this nanomedicine used as eyedrops. HA-RGD conjugation via COOH and amine groups was confirmed by 1H-nuclear magnetic resonance and Fourier-transform infrared spectroscopy. The average diameter of GEH-RGD NPs was 168.87±22.5 nm with positive charge (19.7±2 mV), with an EGCG-loading efficiency up to 95%. Images of GEH-RGD NPs acquired from transmission electron microscopy showed a spherical shape and shell structure of about 200 nm. A slow-release pattern was observed in the nanoformulation at about 30% after 30 hours. Surface plasmon resonance confirmed that GEH-RGD NPs specifically bound to the integrin αvβ3. In vitro cell-viability assay showed that GEH-RGD efficiently inhibited HUVEC proliferation at low EGCG concentrations (20 μg/mL) when compared with EGCG or non-RGD-modified NPs. Furthermore, GEH-RGD NPs significantly inhibited HUVEC migration down to 58%, lasting for 24 hours. In the corneal NV mouse model, fewer and thinner vessels were observed in the alkali-burned cornea after treatment with GEH-RGD NP eyedrops. Overall, this study indicates that GEH-RGD NPs were successfully developed and synthesized as an inhibitor of vascular endothelial cells with specific targeting capacity. Moreover, they can be used in eyedrops to inhibit angiogenesis in corneal NV mice.


Detection and phasing of single base de novo mutations in biopsies from human in vitro fertilized embryos by advanced whole-genome sequencing.
Currently, the methods available for preimplantation genetic diagnosis (PGD) of in vitro fertilized (IVF) embryos do not detect de novo single-nucleotide and short indel mutations, which have been shown to cause a large fraction of genetic diseases. Detection of all these types of mutations requires whole-genome sequencing (WGS). In this study, advanced massively parallel WGS was performed on three 5- to 10-cell biopsies from two blastocyst-stage embryos. Both parents and paternal grandparents were also analyzed to allow for accurate measurements of false-positive and false-negative error rates. Overall, >95% of each genome was called. In the embryos, experimentally derived haplotypes and barcoded read data were used to detect and phase up to 82% of de novo single base mutations with a false-positive rate of about one error per Gb, resulting in fewer than 10 such errors per embryo. This represents a ∼ 100-fold lower error rate than previously published from 10 cells, and it is the first demonstration that advanced WGS can be used to accurately identify these de novo mutations in spite of the thousands of false-positive errors introduced by the extensive DNA amplification required for deep sequencing. Using haplotype information, we also demonstrate how small de novo deletions could be detected. These results suggest that phased WGS using barcoded DNA could be used in the future as part of the PGD process to maximize comprehensiveness in detecting disease-causing mutations and to reduce the incidence of genetic diseases.


Information needs on precision medicine: a survey of Italian health care professionals.
Despite advances in technology development for precision medicine, obstacles remain as barriers to progress and change. In this context simple questions arise: what is the level of understanding of precision medicine among healthcare professionals? We tried to address this question with a survey whose objective was to explore the perception and understanding of precision medicine. A questionnaire was administered to a sample made of oncologists, clinical and hospital pharmacists, pharmacologists and infectiologists in the context of five different Italian national congresses. Participation in the survey was voluntary and anonymous. The questionnaire was filled-in by a total number of 1113 professionals out of 3670 registered participants in the congresses. About half of respondents stated they were not sufficiently informed about precision medicine, and infectiologists were the ones who felt less informed. Most respondents agreed with the basic principles and definitions of precision medicine and believed this new approach is going to require deep changes in healthcare. Our findings show that healthcare professionals have partial knowledge on this topic and that there is a significant association between respondents' knowledge and their clinical specialty. However, despite some misconceptions about precision medicine, a genuine interest and familiarity with its basic principles seems to emerge.


Patency of arterial repairs from wartime extremity vascular injuries.
Extremity vascular injury (EVI) causes significant disability in Veterans of the Afghanistan/Iraq conflicts. Advancements in acute trauma care improved survival and decreased amputations. The study of wartime EVI has relied on successful limb salvage as a surrogate for vascular repair. We used imaging studies as a specific measure of arterial repair durability. Service members with EVI were identified using the Department of Defense Trauma Registry and validated by chart abstraction. Inclusion criteria for the arterial patency subgroup included an initial repair attempt with subsequent imaging reports (duplex ultrasound, CT angiography, and angiogram) documenting initial patency. The cohort of 527 included 140 Veterans with available imaging studies for 143 arterial repairs; median follow-up from injury time to last available imaging study was 19 months (Q1-Q3: 3-58; range: 1-175). Injury mechanism was predominantly explosions (52%) and gunshot wounds (42%). Of the 143 arterial repairs, 81% were vein grafts. Eight repairs were occluded, replaced or included in extremity amputations. One upper extremity and three transtibial late amputations were performed for chronic pain and poor function averaging 27 months (SD: 4; range: 24-32). Kaplan-Meier analysis estimated patency rates of 99%, 97%, 95%, 91% and 91% at 3, 6, 12, 24, and 36 months, respectively, with similar results for upper and lower extremity repairs. Explosive and gunshot wound injury mechanisms had similar patency rates and upper extremity injuries repaired with vein grafts had increased patency. Arterial repair mid-term patency in combat-related extremity injuries is excellent based on imaging studies for 143 repairs. Assertive attempts at acute limb salvage and vascular repair are justified with decisions for amputation versus limb salvage based on the overall condition of the patient and degree of concomitant nerve, orthopedic and soft tissue injuries rather than the presence of arterial injuries. Therapeutic/care management, level IV.


Metabolic response to starvation. II. Effects of sex steroid administration to pre- and postmenopausal women.
Premenopausal women become more ketotic than do men during short term starvation. The possible influence of female sex steroids on ketosis during an 84-hr fast was studied in premenopausal women receiving combined oral contraceptive steroids. The results were compared to age and weight-matched women on no steroid birth control regimen. Although concentrations of several plasma substrates, insulin, and glucagon were measured throughout the fast, no differences between groups were observed with the exception of the 36-hr interval. At this time total plasma ketones and betahydroxybutyrate were significantly higher in women receiving oral contraceptives. In a second study six postmenopausal women were investigated in a similar fashion during a 36-hr fast. The procedure was repeated following a 5 day treatment regimen of oral estradiol and medroxyprogesterone acetate. After sex steroid treatment, total plasma ketones and betahydroxybutyrate concentrations exceeded control values significantly at 36 hr while plasma alanine and free fatty acid levels were significantly lower. The results suggest that oral estrogen-progestin combinations increase the ketogenic response of pre- and postmenopausal women during the early phases of total starvation. This hormonal effect may explain, in part, why women are more sensitive to fasting than are men.


Prospective validation of the Bleeding Academic Research Consortium classification in the all-comer PRODIGY trial.
The Bleeding Academic Research Consortium (BARC) classification has been proposed by consensus to standardize bleeding endpoint definition and reporting in cardiovascular clinical trials. There are no prospective studies on its prognostic impact. We explored the association of BARC-defined bleeding with mortality and compared its prognostic value against two validated bleeding scales: the Thrombolysis in Myocardial Infarction (TIMI) and the Global Utilization of Streptokinase and Tissue Plasminogen Activator for Occluded Coronary Arteries (GUSTO) scales. Non-coronary artery bypass graft (CABG)-related bleedings within the PRODIGY trial were prospectively adjudicated by a blinded Clinical Event Committee and analysed according to multiple statistical modelling. At 2 years, bleeding occurred in 143 patients (7.1%) according to BARC Type 2, 3, or 5; in 50 patients (2.5%) according to TIMI minor or major; and in 61 patients (3.1%) according to GUSTO moderate or severe. One hundred sixty-three patients died (8.1%). After multivariable modelling, BARC Type 2, 3, or 5 bleeding was associated with increased 2-year mortality [hazard ratio (HR): 3.77; 95% confidence interval (CI): 2.37-5.98]. Bleeding Academic Research Consortium Type 3 or 5 was associated with an increased mortality rate at 2 years (adjusted HR: 7.72; 95% CI: 4.75-12.54) similar to that provided by TIMI (HR: 7.64, 95% CI: 4.53-12.87) or GUSTO (HR: 7.36, 95% CI: 4.38-12.34) criteria. In a contemporary, all-comer percutaneous coronary intervention trial actionable BARC bleedings were associated with increased risk of mortality with BARC Type 3 or 5 bleedings providing a similar mortality risk to that posed by TIMI or GUSTO scales.


Fruit and vegetable intake and risk of cardiovascular disease: the Women's Health Study.
Prospective data relating fruit and vegetable intake to cardiovascular disease (CVD) risk are sparse, particularly for women. In a large, prospective cohort of women, we examined the hypothesis that higher fruit and vegetable intake reduces CVD risk. In 1993 we assessed fruit and vegetable intake among 39876 female health professionals with no previous history of CVD or cancer by use of a detailed food-frequency questionnaire. We subsequently followed these women for an average of 5 y for incidence of nonfatal myocardial infarction (MI), stroke, percutaneous transluminal coronary angioplasty, coronary artery bypass graft, or death due to CVD. During 195647 person-years of follow-up, we documented 418 incident cases of CVD including 126 MIs. After adjustment for age, randomized treatment status, and smoking, we observed a significant inverse association between fruit and vegetable intake and CVD risk. For increasing quintiles of total fruit and vegetable intake (median servings/d: 2. 6, 4.1, 5.5, 7.1, and 10.2), the corresponding relative risks (RRs) were 1.0 (reference), 0.78, 0.72, 0.68, and 0.68 (95% CI comparing the 2 extreme quintiles: 0.51, 0.92; P: for trend = 0.01). An inverse, though not statistically significant, trend remained after additional adjustment for other known CVD risk factors, with RRs of 1.0, 0.75, 0.83, 0.80, and 0.85 (95% CI for extreme quintiles: 0.61, 1.17). After excluding participants with a self-reported history of diabetes, hypertension, or high cholesterol at baseline, the multivariate-adjusted RR was 0.45 when extreme quintiles were compared (95% CI: 0.22, 0.91; P: for trend = 0.09). Higher fruit and vegetable intake was also associated with a lower risk of MI, with an adjusted RR of 0.62 for extreme quintiles (95% CI: 0.37, 1.04; P: for trend = 0.07). These data suggest that higher intake of fruit and vegetables may be protective against CVD and support current dietary guidelines to increase fruit and vegetable intake.


Juvenile idiopathic arthritis-associated uveitis at an Italian tertiary referral center: clinical features and complications.
To describe the frequencies and risk factors of ocular complications and poor visual outcomes in children with juvenile idiopathic arthritis (JIA). Retrospective cohort study, including 69 consecutive children (116 eyes) affected by JIA-associated uveitis managed at a tertiary uveitis clinic. The incidence of visual loss to the 20/50 or worse threshold was 0.04/eye-year (EY) and to the 20/200 or worse threshold was 0.02/EY. The most common complications at baseline were posterior synechiae (52%), band keratopathy (38%), and cataract (12%). Risk factor for a visual acuity threshold of 20/50 or worse included hypotony (p = 0.01; hazard ratio [HR] 3.7; 95% CI 1.3-10.4); anterior chamber flare >1 (p = 0.04; HR 1.3; 95% CI 0.5-3.4); a positive antinuclear antibody (ANA) (p = 0.02; HR1.4; 95% CI 0.8-2.4). Hypotony and positive ANA are also associated to the 20/200 or worse threshold (p = 0.03; HR 5.1; 95% CI 1.1-23.9 and p = 0.04; HR 1.0; 95% CI 0.4-2.3; respectively). Use of immunosuppressive drugs was associated with a reduced risk of visual loss of 20/200 or worse (odds ratio 0.14, 95% CI, 0.02-1.29; p = 0.04). Loss of vision and ocular complications still occur among children with JIA-related uveitis. Prompt diagnosis and a strict follow up associated to immunosuppressive therapy may decrease the poor visual outcome.


Epigenetic regulation of melatonin receptors in neuropsychiatric disorders.
Melatonin, the primary indoleamine hormone of the mammalian pineal gland, is known to have a plethora of neuroregulatory, neuroprotective and other properties. Melatonergic signalling is mediated by its two GPCRs, MT1 and MT2 , which are widely expressed in the mammalian CNS. Melatonin levels and receptor expression often show a decrease during normal ageing, and this reduction may be accelerated in some disease states. Depleted melatonergic signalling has been associated with neuropsychiatric dysfunction and impairments in cognition, memory, neurogenesis and neurorestorative processes. The anticonvulsant and mood stabilizer, valproic acid (VPA), up-regulates melatonin MT1 and/or MT2 receptor expression in cultured cells and in the rat brain. VPA is known to affect gene expression through several mechanisms, including the modulation of intracellular kinase pathways and transcription factors, as well as the inhibition of histone deacetylase (HDAC) activity. Interestingly, other HDAC inhibitors, such as trichostatin A, which are structurally distinct from VPA, can also up-regulate melatonin receptor expression, unlike a VPA analogue, valpromide, which lacks HDAC inhibitory activity. Moreover, VPA increases histone H3 acetylation along the length of the MT1 gene promoter in rat C6 cells. These findings indicate that an epigenetic mechanism, linked to histone hyperacetylation/chromatin remodelling and associated changes in gene transcription, is involved in the up-regulation of melatonin receptors by VPA. Epigenetic induction of MT1 and/or MT2 receptor expression, in areas where these receptors are lost because of ageing, injury or disease, may be a promising therapeutic avenue for the management of CNS dysfunction and other disorders. LINKED ARTICLES: This article is part of a themed section on Recent Developments in Research of Melatonin and its Potential Therapeutic Applications. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.16/issuetoc.


Experimental evaluation of new chitin-chitosan graft for duraplasty.
Natural materials such as collagen and alginate have promising applications as dural graft substitutes. These materials are able to restore the dural defect and create optimal conditions for the development of connective tissue at the site of injury. A promising material for biomedical applications is chitosan-a linear polysaccharide obtained by the deacetylation of chitin. It has been found to be nontoxic, biodegradable, biofunctional and biocompatible in addition to having antimicrobial characteristics. In this study we designed new chitin-chitosan substitutes for dura mater closure and evaluated their effectiveness and safety. Chitosan films were produced from 3 % of chitosan (molar mass-200, 500 or 700 kDa, deacetylation rate 80-90%) with addition of 20% of chitin. Antimicrobial effictively and cell viability were analysed for the different molar masses of chitosan. The film containing chitosan of molar mass 200 kDa, had the best antimicrobial and biological activity and was successfully used for experimental duraplasty in an in vivo model. In conclusion the chitin-chitosan membrane designed here met the requirements for a dura matter graft exhibiting the ability to support cell growth, inhibit microbial growth and biodegradade at an appropriate rate. Therefore this is a promising material for clinical duroplasty.


Vector-free intracellular delivery by reversible permeabilization.
Despite advances in intracellular delivery technologies, efficient methods are still required that are vector-free, can address a wide range of cargo types and can be applied to cells that are difficult to transfect whilst maintaining cell viability. We have developed a novel vector-free method that uses reversible permeabilization to achieve rapid intracellular delivery of cargos with varying composition, properties and size. A permeabilizing delivery solution was developed that contains a low level of ethanol as the permeabilizing agent. Reversal of cell permeabilization is achieved by temporally and volumetrically controlling the contact of the target cells with this solution. Cells are seeded in conventional multi-well plates. Following removal of the supernatant, the cargo is mixed with the delivery solution and applied directly to the cells using an atomizer. After a short incubation period, permeabilization is halted by incubating the cells in a phosphate buffer saline solution that dilutes the ethanol and is non-toxic to the permeabilized cells. Normal culture medium is then added. The procedure lasts less than 5 min. With this method, proteins, mRNA, plasmid DNA and other molecules have been delivered to a variety of cell types, including primary cells, with low toxicity and cargo functionality has been confirmed in proof-of-principle studies. Co-delivery of different cargo types has also been demonstrated. Importantly, delivery occurs by diffusion directly into the cytoplasm in an endocytic-independent manner. Unlike some other vector-free methods, adherent cells are addressed in situ without the need for detachment from their substratum. The method has also been adapted to address suspension cells. This delivery method is gentle yet highly reproducible, compatible with high throughput and automated cell-based assays and has the potential to enable a broad range of research, drug discovery and clinical applications.


Long-term regulation of contractility and calcium current in smooth muscle.
Longitudinal smooth muscle strips from guinea pig ileum were cultured in vitro for 5 days, and the relationship between extracellular Ca2+ and force in high-K+ medium was evaluated. In strips cultured with 10% fetal calf serum (FCS), this relationship was shifted to the right (50% effective concentration changed by 2-3 mM) compared with strips cultured without FCS. The shift was prevented by inclusion of verapamil (1 microM) during culture and mimicked by ionomycin in the absence of FCS. The intracellular Ca2+ concentration ([Ca2+]i) during stimulation with high-K+ solution or carbachol was reduced after culture with FCS, whereas the [Ca2+]i-force relationship was unaffected. Cells were isolated from cultured strips, and whole cell voltage-clamp experiments were performed. Maximum inward Ca2+ current (10 mM Ba2+), normalized to cell capacitance, was almost three times smaller in cells isolated from strips cultured with FCS. Culture with 1 microM verapamil prevented this reduction. These results suggest that increased [Ca2+]i during culture downregulates Ca2+ current density, with associated effects on contractility.


Unrelated donor marrow transplantation for inborn errors.
From December 1989 to December 1997 40 children aged 1 year to 19 years with inborn errors other than severe combined immunodeficiencies underwent unrelated donor (UD) bone marrow transplantation (BMT) in one of 10 institutions of the Italian Bone Marrow Transplant Group participating in this program. The diseases leading to BMT included Fanconi Anemia (10), Thalassemia (8), Wiskott Aldrich syndrome (5), haemophagocytic lymphohystiocytosis (6), osteopetrosis (3), storage diseases (6), Chediak Higashi syndrome (1), Schwachman syndrome (1). Thirty-three pairs were A, B, DRB1 matched. Three pairs were one antigen mismatched and one pair was two antigens mismatched. The remaining three pairs lacked information on molecular biology. Twelve children underwent a preparative regimen including radiotherapy. The remaining 28 children were conditioned with a chemotherapy regimen which included Busulfan. GvHD disease prophylaxis included CSA and MTX alone (9) or associated with ALG (17) or in vivo Campath 1G (12). The remaining two children received CSA alone. Thirty-five children showed donor engraftment; three children with thalassemia and one with osteopetrosis failed to engraft. Five children developed secondary graft failure. Actuarial 5 year disease-free survival was 62%; grade III-IV acute GvHD developed in seven of 38 evaluable children (18%); chronic GvHD developed in seven of 27 evaluable children (26%). We confirm that Wiskott Aldrich syndrome, HLH, and osteopetrosis represent an absolute indication for UD-BMT. Prognosis of UD-BMT for FA could improve in children grafted in an early phase, but a better preparative regimen has to be identified. UD-BMT in thalassemia is acceptable only in a restricted subset of patients selected for poor compliance to therapy.


Forecasting intraspecific changes in distribution of a wide-ranging marine predator under climate change.
Globally, marine animal distributions are shifting in response to a changing climate. These shifts are usually considered at the species level, but individuals are likely to differ in how they respond to the changing conditions. Here, we investigate how movement behaviour and, therefore, redistribution, would differ by sex and maturation class in a wide-ranging marine predator. We tracked 115 tiger sharks (Galeocerdo cuvier) from 2002 to 2020 and forecast class-specific distributions through to 2030, including environmental factors and predicted occurrence of potential prey. Generalised Linear and Additive Models revealed that water temperature change, particularly at higher latitudes, was the factor most associated with shark movements. Females dispersed southwards during periods of warming temperatures, and while juvenile females preferred a narrow thermal range between 22 and 23 °C, adult female and juvenile male presence was correlated with either lower (< 22 °C) or higher (> 23 °C) temperatures. During La Niña, sharks moved towards higher latitudes and used shallower isobaths. Inclusion of predicted distribution of their putative prey significantly improved projections of suitable habitats for all shark classes, compared to simpler models using temperature alone. Tiger shark range off the east coast of Australia is predicted to extend ~ 3.5° south towards the east coast of Tasmania, particularly for juvenile males. Our framework highlights the importance of combining long-term movement data with multi-factor habitat projections to identify heterogeneity within species when predicting consequences of climate change. Recognising intraspecific variability will improve conservation and management strategies and help anticipate broader ecosystem consequences of species redistribution due to ocean warming.


Effect of nitrofurazone on the reproductive organs in adult male mice.
To study the effect of 5-nitro-2-furaldehyde semicarbazone (nitrofurazone), a derivative of nitrofuran, on male reproductive organs of Parkes (P) strain mice. Mice were given nitrofurazone orally at a dose of 64 mg/kg body weight per day, for 10 and 20 days, and were killed 24 h and/or 56 days after the last treatment. Histological appearance of testis, motility and number of spermatozoa in cauda epididymidis, and biochemical indices in epididymis and seminal vesicle were evaluated. Histologically, testis showed marked regressive changes in the seminiferous tubules in mice treated with nitrofurazone. Ten days after treatment, there was much depletion of germ cells in the seminiferous tubules, and the germinal epithelium was lined mainly with Sertoli cells, spermatogonia, spermatocytes, and a few round spermatids; intraepithelial vacuoles and multinucleated giant cells were also observed in tubules. By 20 days, regressive changes in the seminiferous tubules were further pronounced, and pachytene spermatocytes were the most advanced germ cells noticed in the tubules. In severe cases, the tubules were lined with a thin layer of Sertoli cells and spermatogonia. The treatment also caused marked reductions in the motility and number of spermatozoa in the cauda epididymidis, in weight and the level of fructose in the seminal vesicle, and in sialic acid level in the epididymis. Fifty six days after drug withdrawal, the alterations induced in the reproductive organs returned to control levels. Our results suggest that nitrofurazone treatment in P mice induces marked alterations in the male reproductive organs, and that the alterations are reversible following cessation of treatment.


Multicenter evaluation of MRI-based radiomic features: A phantom study.
This work describes the development of a novel radiomics phantom designed for magnetic resonance imaging (MRI) that can be used in a multicenter setting. The purpose of this study is to assess the stability and reproducibility of MRI-based radiomic features using this phantom across different MRI scanners. A set of phantoms were three-dimensional (3D) printed using MRI visible materials. One set of phantoms were imaged on seven MRI scanners and one was imaged on one MRI scanner. Radiomics analysis of the phantoms, which included first-order features, shape and texture features was performed. Intraclass correlation coefficient (ICC) was used to assess the stability of radiomic features across eight scanners and the reproducibility of two printed models on one scanner. Coefficient of variation (COV) was used to assess the reproducibility of radiomics measurements in the phantom on a single scanner. The phantom models provide sufficient signal-to-noise and contrast in all the tumor models permitting robust automatic segmentation. During a 12-month period of monitoring, the phantom material was stable with T1 and T2 of 150.7 ± 6.7 ms and 56.1 ± 3.9 ms, respectively. Of all the radiomic features computed, 34 of 69 had COV < 10%. Features from first-order statistics were the most robust in stability across the eight scanners with eight of 12 (67%) having high stability. About 29 of 50 (58%) texture features had high stability and no shape features had high stability features across the eight scanners. A novel MRI radiomics phantom has been developed to assess the reproducibility and stability of MRI-based radiomic features across multiple institutions. The variation in radiomic feature stability demonstrates the need for caution when interpreting these features for clinical studies.


Changes in the MicroRNA Profile Observed in the Subcutaneous Adipose Tissue of Obese Patients after Laparoscopic Adjustable Gastric Banding.
Background. Laparoscopic adjustable gastric banding (LAGB) results in significant lasting weight loss and improved metabolism in obese patients. To evaluate whether epigenetic factors could concur to these benefits, we investigated the subcutaneous adipose tissue (SAT) microRNA (miRNA) profile before (T0) and three years (T1) after LAGB in three morbidly obese women. Case Reports. SAT miRNA profiling, evaluated by TaqMan Array, showed four downexpressed (miR-519d, miR-299-5p, miR-212, and miR-671-3p) and two upexpressed (miR-370 and miR-487a) miRNAs at T1 versus T0. Bioinformatics predicted that these miRNAs regulate genes belonging to pathways associated with the cytoskeleton, inflammation, and metabolism. Western blot analysis showed that PPAR-alpha, which is the target gene of miR-519d, increased after LAGB, thereby suggesting an improvement in SAT lipid metabolism. Accordingly, the number and diameter of adipocytes were significantly higher and lower, respectively, at T1 versus T0. Bioinformatics predicted that the decreased levels of miR-212, miR-299-5p, and miR-671-3p at T1 concur in reducing SAT inflammation. Conclusion. We show that the miRNA profile changes after LAGB. This finding, although obtained in only three cases, suggests that this epigenetic mechanism, by regulating the expression of genes involved in inflammation and lipid metabolism, could concur to improve SAT functionality in postoperative obese patients.


Effectiveness and Mechanisms of Defluorination of Perfluorinated Alkyl Substances by Calcium Compounds during Waste Thermal Treatment.
The mineralization of perfluorinated alkyl substances (PFASs) by calcium compounds during the waste thermal treatment was systemically studied. Different calcium compounds showed different mineralization efficiencies of PFASs during the thermal process, owing to the different reaction mechanisms. Calcium hydroxide was recommended as the most effective Ca reagent for PFAS defluorination because the carbon-fluorine bonds in PFASs can be converted to carbon-hydrogen bonds via the hydrodefluorination reaction. PFASs with different chain lengths and functional groups were further investigated for their potentially different mineralization behavior. The results showed that the chain length of PFASs had an insignificant effect on the mineralization efficiency by calcium hydroxide. The thermogravimetric analysis-differential thermal analysis (TGA-DTA) also revealed that perfluorooctanesulfonate (PFOS) and perfluorohexanesulfonate (PFHxS) (with different chain lengths) had a similar thermal behavior. However, PFASs with different functional groups showed different mineralization behavior with calcium hydroxide in relation to their different thermal decomposition temperatures. Finally, the mineralization ratio of polytetrafluoroethylene (PTFE) particles by calcium hydroxide could reach 80% or higher when the temperature was above 400 °C. The gas chromatography/mass spectrometry (GC/MS) results demonstrated much reduced production of gaseous fluorocarbon fragments during PTFE decomposition when coexisting with calcium hydroxide.


Non-monotonous enzyme-assisted self-assembly profiles resulting from reaction-diffusion processes in host gels.
Reaction-diffusion (RD) processes are responsible for surface and in-depth micropatterning in inanimate and living matter. Here we show that enzyme-assisted self-assembly (EASA) of peptides is a valuable tool to functionnalize host gels. By using a phosphatase distributed in a host hydrogel, the diffusion of phosphorylated peptides from a liquid/host gel interface leads to the spontaneous formation of a pattern of dephosphorylated peptide self-assembly presenting at least two self-assembly maxima. Variation of enzyme and peptide concentrations change the pattern characteristics. When a peptide drop is deposited on a phosphatase functionalized gel, a self-assembly pattern is also formed both along the gel-solution interface and perpendicular to the interface. This self-assembly pattern induces a local change of the gel mechanical properties measured by nanoindentation. Its appearance relies on the formation of self-assembled structures by nucleation and growth processes which are static in the hydrogel. This process presents great similarities with the Liesegang pattern formation and must be taken into account for the functionalization of hydrogels by EASA. A mechanism based on RD is proposed leading to an effective mathematical model accounting for the pattern formation. This work highlights EASA as a tool to design organic Liesegang-like microstructured materials with potential applications in biomaterials and artificial living systems design.


Indications for laparoscopic surgery for Crohn's disease using the Vienna Classification.
The aim of this study was to investigate the clinical outcome of laparoscopic surgery for Crohn's disease and clarify the indications using the Vienna Classification. Between September 1994 and July 2004, 107 patients with Crohn's disease underwent 124 procedures. Of these, 91 laparoscopic procedures formed the basis of this study. The Vienna Classification, which consists of three subgroups - age at diagnosis (A1-2), location (L1-4) and behaviour (B1-3) - was applied to compare the conversion to open surgery and incidence of postoperative complications. Conversion to open surgery was necessary in 12 (13.2%) patients. Major and minor postoperative complications occurred in five (5.5%) and 13 (19.8%) patients respectively. The conversion rate, major and total complications in the B3L3/4 subgroup were significantly greater than in the other subgroups. Multivariate analysis showed that B3L3/4 was the only predictive factor for all complications. However, the incidence of major and all complications in the B3L3/4 subgroup did not differ between the open and laparoscopic surgery groups. Laparoscopic surgery for Crohn's disease is the procedure of choice for all uncomplicated cases (B2L1-4, B3L1/2). For patients in the complicated group (B3L3/4), laparoscopy is also feasible and justified; however, the surgeon must be aware of the propensity for higher rate of conversion.


Toward a systematic exploration of nano-bio interactions.
Many studies of nanomaterials make non-systematic alterations of nanoparticle physicochemical properties. Given the immense size of the property space for nanomaterials, such approaches are not very useful in elucidating fundamental relationships between inherent physicochemical properties of these materials and their interactions with, and effects on, biological systems. Data driven artificial intelligence methods such as machine learning algorithms have proven highly effective in generating models with good predictivity and some degree of interpretability. They can provide a viable method of reducing or eliminating animal testing. However, careful experimental design with the modelling of the results in mind is a proven and efficient way of exploring large materials spaces. This approach, coupled with high speed automated experimental synthesis and characterization technologies now appearing, is the fastest route to developing models that regulatory bodies may find useful. We advocate greatly increased focus on systematic modification of physicochemical properties of nanoparticles combined with comprehensive biological evaluation and computational analysis. This is essential to obtain better mechanistic understanding of nano-bio interactions, and to derive quantitatively predictive and robust models for the properties of nanomaterials that have useful domains of applicability.


Governance regime and location influence avoided deforestation success of protected areas in the Brazilian Amazon.
Protected areas in tropical countries are managed under different governance regimes, the relative effectiveness of which in avoiding deforestation has been the subject of recent debates. Participants in these debates answer appeals for more strict protection with the argument that sustainable use areas and indigenous lands can balance deforestation pressures by leveraging local support to create and enforce protective regulations. Which protection strategy is more effective can also depend on (i) the level of deforestation pressures to which an area is exposed and (ii) the intensity of government enforcement. We examine this relationship empirically, using data from 292 protected areas in the Brazilian Amazon. We show that, for any given level of deforestation pressure, strictly protected areas consistently avoided more deforestation than sustainable use areas. Indigenous lands were particularly effective at avoiding deforestation in locations with high deforestation pressure. Findings were stable across two time periods featuring major shifts in the intensity of government enforcement. We also observed shifting trends in the location of protected areas, documenting that between 2000 and 2005 strictly protected areas were more likely to be established in high-pressure locations than in sustainable use areas and indigenous lands. Our findings confirm that all protection regimes helped reduce deforestation in the Brazilian Amazon.


IgA production in MHC class II-deficient mice is primarily a function of B-1a cells.
Mice deficient in MHC class II expression (C2d mice) do not make antibody to protein antigens administered systemically, but their ability to produce IgA antibody to antigen administered at mucosal sites has not been described. We investigated IgA production by C2d mice and their IgA antibody response to antigen given orally. Young C2d mice had normal amounts of serum IgA, intestinal-secreted IgA and normal numbers of intestinal IgA plasma cells, compared to control C57BL/6 mice. IgA production by C2d mice increased with age. Following oral immunization with cholera toxin, C57BL/6 mice responded with IgA and IgG antibody, and had increased numbers of IgA plasma cells, but C2d mice gave no response. The Peyer's patch and mesenteric lymph node tissues of C2d mice contained very few CD4-expressing T cells. Thus, C2d mice have no typical mucosal CD4 Th cells and cannot respond to a strong oral immunogen, yet they still produced and secreted IgA. We hypothesized that B-1 lymphocytes could provide a source of IgA independent of antigen-specific T cell help. Young C2d mice had normal numbers of peritoneal B-1a cells and their frequency increased with age. To test the role of these B-1a cells, we bred C2d mice to obtain mice that had no MHC class II expression and expressed the Xid gene that confers deficiency in B-1a cells. These double-deficient mice had 10-fold less serum and secreted IgA than all other F2 littermates. We conclude that B-1a cells are essential for the majority of IgA production in C2d mice. Thus, the C2d mouse may provide a useful tool for analysis of the role of intestinal IgA provided by B-1a cells.


Scabies in some workers living in crowded area, Jeddah, Saudi Arabia.
Little is known about human and animal scabies in Jeddah. The present study gives a picture of human scabies. Over four months, 18 cases have been microscopically diagnosed. The most common infested site was the abdomen, followed by the lower and upper limbs. Infestation in other parts of the body was not common but also recorded. The results were discussed on the light of other works done in the region.


The Women's Interagency HIV Study. WIHS Collaborative Study Group.
The Women's Interagency HIV Study comprises the largest U.S. cohort to date of human immunodeficiency virus (HIV)-seropositive women (N = 2,058) with a comparison cohort of seronegative women (N = 568). The methodology, training, and quality assurance activities employed are described. The study population, enrolled between October 1994 and November 1995 through six clinical consortia throughout the United States (totaling 23 sites) represents a typically hard-to-reach study population. More than half of the women in each cohort were living below the federally defined levels of poverty. The women ranged in age from 16 to 73 years; approximately one-quarter self-identified as Latina or Hispanic, over one-half as African-American not of Hispanic origin, and less than 20% as white, non-Hispanic origin. Self-reporting of HIV exposure risk included injection drug use by 34% of the seropositive women and 28% of the seronegative women, heterosexual contact (42% vs 26%), transfusion risk (4% vs 3%) and no identified risk (20% vs 43%). Demographic and HIV exposure risk characteristics of the seropositive cohort were comparable with characteristics of nationally reported AIDS cases in U.S. women. This well characterized cohort of HIV-seropositive and high-risk seronegative women represents a rich opportunity for future studies of HIV disease progression and pathogenesis.