Intracardiac thrombus in children: the fine equilibrium between the risk and the benefit.
The medical records of 16 patients diagnosed as intracardiac thrombus were searched. The size, location and outcome of thrombus together with demographic data of patients were assessed. The median age of the patients was 2.2 years. Six patients were newborn and two patients were infant. The median size of thrombus was 9 mm. The localization was right atrium in seven, right ventricle in five, left ventricle in one, pulmonary artery in one, and superior vena cava in two patients. There was prematurity in five, ciyanotic congenital heart disease in one, blood culture positivity in three, malignancy in four, nephrotic syndrome in one, indwelling catheters in 10, and acquired or genetic thrombophilia in six patients as risk factors. In the treatment, the first choice was tissue plasminogen activator in two patients, heparin infusion in one patient and low molecular weight heparin in remaining 12 patients. In nine patients, therapy included parenteral antimicrobials together with anticoagulants. The result was complete resolution in 15 patients and in one patient thrombus was surgically removed. The median time was 16 (2-70) days for 50% resolution and 26 (3-93) days for complete resolution. There was a statistically significant (P = .027 and r = 0.5) correlation between the size and the complete resolution time. There was no anticoagulant therapy related major complication. In patients with intracardiac thrombus, selection of anticoagulant therapy may decrease the risk of complications. Surgery is rarely required and thrombolytics are not usually necessary for resolution of thrombus.


Glutamate regulates Ca2+ signals in smooth muscle cells of newborn piglet brain slice arterioles through astrocyte- and heme oxygenase-dependent mechanisms.
Glutamate is the principal cerebral excitatory neurotransmitter and dilates cerebral arterioles to match blood flow to neural activity. Arterial contractility is regulated by local and global Ca(2+) signals that occur in smooth muscle cells, but modulation of these signals by glutamate is poorly understood. Here, using high-speed confocal imaging, we measured the Ca(2+) signals that occur in arteriole smooth muscle cells of newborn piglet tangential brain slices, studied signal regulation by glutamate, and investigated the physiological function of heme oxygenase (HO) and carbon monoxide (CO) in these responses. Glutamate elevated Ca(2+) spark frequency by approximately 188% and reduced global intracellular Ca(2+) concentration ([Ca(2+)](i)) to approximately 76% of control but did not alter Ca(2+) wave frequency in brain arteriole smooth muscle cells. Isolation of cerebral arterioles from brain slices abolished glutamate-induced Ca(2+) signal modulation. In slices treated with l-2-alpha-aminoadipic acid, a glial toxin, glutamate did not alter Ca(2+) sparks or global [Ca(2+)](i) but did activate Ca(2+) waves. This shift in Ca(2+) signal modulation by glutamate did not occur in slices treated with d-2-alpha-aminoadipic acid, an inactive isomer of l-2-alpha-aminoadipic acid. In the presence of chromium mesoporphyrin, a HO blocker, glutamate inhibited Ca(2+) sparks and Ca(2+) waves and did not alter global [Ca(2+)](i). In isolated arterioles, CORM-3 [tricarbonylchloro(glycinato)ruthenium(II)], a CO donor, activated Ca(2+) sparks and reduced global [Ca(2+)](i). These effects were blocked by 1H-(1,2,4)-oxadiazolo-(4,3-a)-quinoxalin-1-one, a soluble guanylyl cyclase inhibitor. Collectively, these data indicate that glutamate can modulate Ca(2+) sparks, Ca(2+) waves, and global [Ca(2+)](i) in arteriole smooth muscle cells via mechanisms that require astrocytes and HO. These data also indicate that soluble guanylyl cyclase is involved in CO activation of Ca(2+) sparks in arteriole smooth muscle cells.


Rapid de novo evolution of lysis genes in single-stranded RNA phages.
Leviviruses are bacteriophages with small single-stranded RNA genomes consisting of 3-4 genes, one of which (sgl) encodes a protein that induces the host to undergo autolysis and liberate progeny virions. Recent meta-transcriptomic studies have uncovered thousands of leviviral genomes, but most of these lack an annotated sgl, mainly due to the small size, lack of sequence similarity, and embedded nature of these genes. Here, we identify sgl genes in 244 leviviral genomes and functionally characterize them in Escherichia coli. We show that leviviruses readily evolve sgl genes and sometimes have more than one per genome. Moreover, these genes share little to no similarity with each other or to previously known sgl genes, thus representing a rich source for potential protein antibiotics.


CD4 T cells and major histocompatibility complex class II expression influence worm expulsion and increased intestinal muscle contraction during Trichinella spiralis infection.
Expulsion of intestinal nematode parasites and the associated increased contraction by intestinal muscle are T cell dependent, since both are attenuated in athymic rodents. The CD4 T-cell subset has been strongly associated with worm expulsion; however, the relationship between these cells, antigen presentation, and worm expulsion is not definitive and the role of these factors in intestinal muscle hypercontractility has not been defined. We infected C57BL/6, athymic, CD4-deficient, CD8alpha-deficient, and major histocompatibility complex class II (MHC II)-deficient (C2d) mice with Trichinella spiralis larvae. We examined intestinal worm numbers, longitudinal muscle contraction, and MHC II expression. Numerous MHC II-positive cells were identified within the muscularis externa of infected but not uninfected C57BL/6 mice. C57BL/6 and CD8alpha-deficient mice developed large increases in muscle contraction, expelling the parasite by day 21. Athymic and C2d mice exhibited much smaller increases in muscle contraction and delayed parasite expulsion. CD4-deficient mice exhibited intermediate levels of muscle contraction and delayed parasite expulsion. To further examine the role of MHC II and CD4 T cells, we irradiated C2d mice and reconstituted them with C57BL/6 bone marrow alone or with C57BL/6 CD4 T cells. C57BL/6 bone marrow alone did not affect muscle function or worm expulsion in recipient C2d mice. Partial CD4 T-cell reconstitution was sufficient to restore increased muscle contraction but not worm expulsion. Thus, hematopoietic MHC II expression alone is insufficient for the development of muscle hypercontractility and worm expulsion, but the addition of even small numbers of CD4 T cells was sufficient to induce intestinal muscle pathophysiology.


Need assessment for HIV drug resistance testing and landscape of current and future technologies in low- and middle-income countries.
Resistance to antiretroviral drugs used to treat HIV is an important and evolving concern, particularly in low- and middle-income countries (LMICs) which have been impacted to the greatest extent by the HIV pandemic. Efforts to monitor the emergence and transmission of resistance over the past decade have shown that drug resistance-especially to the nucleoside analogue and non-nucleoside reverse transcriptase inhibitors-can (and have) increased to levels that can jeopardize the efficacy of available treatment options at the population level. The global shift to integrase-based regimens as the preferred first-line therapy as well as technological advancements in the methods for detecting resistance have had an impact in broadening and diversifying the landscape of and use case for HIV drug resistance testing. This review estimates the potential demand for HIV drug resistance tests, and surveys current testing methodologies, with a focus on their application in LMICs.


Cooked broiler meat quality affected by different Mediterranean medicinal plants in the diet.
This study was conducted to investigate the effects of adding oregano (Origanum syriacum L.) and germander (Teucrium polium L.) to poultry diets individually and/or in combination: i) on cooked chicken meat quality and storage stability, ii) to compare this effect with those of the synthetic antioxidant butylated hydroxyanisole (BHA) and with the normal basic diet (Control: without supplements). Broilers (140 birds) were raised for 21 days and then equally divided into five different treatment groups of 28 birds each. The dietary treatments were as follows: i) control; ii) germander (GER, 1.5%); iii) oregano (ORE, 2.5%); iv) combination of GER and ORE (CM, 1.5%, and 2.5%); v) BHA (0.02%). Meat patties from the five treatments were prepared, cooked, and stored at 4°C prior to analysis. During storage, samples were measured for thiobarbituric acid-reactive substances (TBARS) and total carbonyl levels at 0, 4, and 7 days. In addition, cooked thigh meat was prepared separately to evaluate cooking loss and sensory attributes. The CM dietary treatment showed the highest antioxidant effect, with decreasing TBARS values (breast and thigh meat) throughout the storage time (4 to 7 days). Furthermore, ORE showed a higher antioxidant effect, decreasing the rancidity development (TBARS values), compared to the GER during the storage period (days 0 to 7). The anticarbonyl effect of the CM supplement was the highest among all treatments from day 0 to 7. Generally, the antioxidant effect of GER was lower compared to that of ORE and BHA alone. The CM treatment most significantly decreased off-odor and rancidity development, with the lowest oxidation odor scores. The results indicate that the combination of oregano and germander in the diet of boilers improves meat quality and prolongs shelf life.


Oral health-related cultural beliefs for four racial/ethnic groups: Assessment of the literature.
The purpose of this study was to assess information available in the dental literature on oral health-related cultural beliefs. In the US, as elsewhere, many racial/ethnic minority groups shoulder a disproportionate burden of oral disease. Cultural beliefs, values and practices are often implicated as causes of oral health disparities, yet little is known about the breadth or adequacy of literature about cultural issues that could support these assertions. Hence, this rigorous assessment was conducted of work published in English on cultural beliefs and values in relation to oral health status and dental practice. Four racial/ethnic groups in the US (African-American, Chinese, Filipino and Hispanic/Latino) were chosen as exemplar populations. The dental literature published in English for the period 1980-2006 noted in the electronic database PUBMED was searched, using keywords and MeSH headings in different combinations for each racial/ethnic group to identify eligible articles. To be eligible the title and abstract when available had to describe the oral health-related cultural knowledge or orientation of the populations studied. Overall, the majority of the literature on racial/ethnic groups was epidemiologic in nature, mainly demonstrating disparities in oral health rather than the oral beliefs or practices of these groups. A total of 60 relevant articles were found: 16 for African-American, 30 for Chinese, 2 for Filipino and 12 for Hispanic/Latino populations. Data on beliefs and practices from these studies has been abstracted, compiled and assessed. Few research-based studies were located. Articles lacked adequate identification of groups studied, used limited methods and had poor conceptual base. The scant information available from the published dental and medical literature provides at best a rudimentary framework of oral health related ideas and beliefs for specific populations.


Functional connectivity dynamics among cortical neurons: a dependence analysis.
This paper quantifies and comparatively validates functional connectivity between neurons by measuring the statistical dependence between their firing rates. Based on statistical analysis of the pairwise functional connectivity, we estimate, exclusively from neural data, the neural assembly functional connectivity given a behavior task, which provides a quantifiable representation of the dynamic nature during the behavioral task. Because of the time scale of behavior (100-1000 ms), a statistical method that yields robust estimators for this small sample size is desirable. In this work, the temporal resolutions of four estimators of functional connectivity are compared on both simulated data and real neural ensemble recordings. The comparison highlights how the properties and assumptions of statistical-based and phase-based metrics affect the interpretation of connectivity. Simulation results show that mean square contingency (MSC) and mutual information (MI) create more robust quantification of functional connectivity under identical conditions than cross correlation (CC) and phase synchronization (PhS) when the sample size is 1 s. The results of the simulated analysis are extended to real neuronal recordings to assess the functional connectivity in monkey's cortex corresponding to three movement states in a food reaching task and construct the assembly graph given a movement state and the activation degree of a state-related assembly over time using the statistical test exclusively from neural data dependencies. The activation degree of a given state-related assembly reaches the peak repeatedly when the specific movement states occur, which also reveals the network of interactions among the neurons are key for the operation of a specific behavior.


Molecular dynamics study of alkylsilane monolayers on realistic amorphous silica surfaces.
Interfacial properties of n-alkylsilane monolayers on silica have been investigated with molecular dynamics simulations using both reactive and classical (i.e., nonreactive) force fields. A synthesis mimetic simulation (SMS) procedure using the reactive force field ReaxFF has been developed to mimic the experimental processing of silicon wafers involved in the preparation of alkylsilane monolayers; in the SMS procedure, amorphous silica surfaces are generated and exposed to hydrogen peroxide (H2O2) to create a hydroxide surface layer. Alkylsilane monolayers are then assembled on these surfaces, and their behavior is studied. To investigate the impact of the SMS procedure on monolayer properties, simulations have also been performed using more idealized monolayers assembled on crystalline surfaces and non-H2O2-processed amorphous surfaces. The simulations reported here demonstrate that processing-induced silica surface roughness plays a key role in the structure and frictional performance of monolayers. Furthermore, ignoring these effects results in a significant underestimation of the coefficient of friction and an overestimation of the orientational ordering of the monolayers.


The evolution of Candida species and fluconazole susceptibility among oral and vaginal isolates recovered from human immunodeficiency virus (HIV)-seropositive and at-risk HIV-seronegative women.
Antifungal agents can effectively treat mucosal candidiasis; however, their use can lead to colonization with less susceptible species and to resistance among normally susceptible strains. Oral and vaginal Candida isolates obtained at 3 points over 2 years from human immunodeficiency virus (HIV)-seropositive and at-risk HIV-seronegative women were identified by species and were evaluated for in vitro fluconazole susceptibility. Prevalence of non-C. albicans strains increased over time, and these strains were more likely among women reporting current antifungal use. Among C. albicans isolates, resistance was rare, with no evidence for progressive reduction in susceptibility over time. Among non-C. albicans isolates, reduced susceptibility occurred frequently and increased with time. HIV-seropositive women were more likely to have non-C. albicans isolates with reduced susceptibility as were women reporting current antifungal use. This evolution and section of mucosa-colonizing Candida species with reduced susceptibility could play a critical early role in the development of antifungal resistance among C. albicans isolates responsible for refractory candidiasis.


Effect of CT Reconstruction Algorithm on the Diagnostic Performance of Radiomics Models: A Task-Based Approach for Pulmonary Subsolid Nodules.
We investigated whether the diagnostic performance of machine learning-based radiomics models for the discrimination of invasive pulmonary adenocarcinomas (IPAs) among subsolid nodules (SSNs) was affected by the proportion of images reconstructed with filtered back projection (FBP) and model-based iterative reconstruction (MBIR) in datasets used for feature extraction. This retrospective study included 60 patients (23 men and 37 women; mean age, 61.4 years) with 69 SSNs (54 part-solid and 15 pure ground-glass nodules). Preoperative CT scans were reconstructed with both FBP and MBIR. A total of 860 radiomics features were obtained from the entire nodule volume, and 70 resampled nodule datasets with an increasing proportion of nodules with MBIR-derived features (from 0/69 to 69/69) were prepared. After feature selection using neighborhood component analysis, support vector machines (SVMs) and an ensemble model were used as classifiers for the differentiation of IPAs. The diagnostic performances of all blending proportions of reconstruction algorithms were calculated and analyzed. The ROC AUC and the diagnostic accuracy of the radiomics models decreased significantly as the number of nodules with MBIR-derived features increased, and this relationship followed cubic functions (R2 = 0.993 and 0.926 for SVM; R2 = 0.993 and 0.975 for the ensemble model; p < 0.001). The magnitude of variation in AUC due to the reconstruction algorithm heterogeneity was 0.39 for SVM and 0.39 for the ensemble model. Inclusion of CT scans reconstructed with MBIR for radiomics modeling can significantly decrease diagnostic performance for the identification of IPAs.


Loss of the transmembrane but not the soluble kit ligand isoform increases testicular germ cell tumor susceptibility in mice.
Several genetic variants act as modifiers of testicular germ cell tumor (TGCT) susceptibility in the 129/Sv mouse model of human pediatric TGCTs. One such modifier, the Steel locus, encodes the transmembrane-bound and soluble ligand of the kit receptor. Some (Sl and SlJ) but not all (Sld) mutations of the Steel locus increase TGCT incidence in heterozygous mutant mice. Because Sl and SlJ are large deletions that affect multiple transcripts and Sld is an intragenic deletion of the kit ligand (Kitl) from which only the soluble protein is produced, it was uncertain whether Kitl or a neighboring gene is a modifier of TGCT susceptibility. We tested the effect of the small Steel grizzle-belly (Slgb) deletion on TGCT susceptibility to determine whether Kitl is a TGCT modifier gene. An increase in TGCT incidence was observed in Slgb/+ heterozygotes, and fine mapping of the deletion breakpoints revealed that Kitl is the only conventional gene deleted by the mutation, suggesting that Kitl is the TGCT modifier gene at the Steel locus. Additionally, we propose that soluble KITL in Sld/+ heterozygous mutant mice complements a dosage effect of transmembrane-associated kit ligand on TGCT susceptibility and that the kit receptor (Kit) is haplosufficient for primordial germ cell development.


Chitosan-coated liposomes loaded with butyric acid demonstrate anticancer and anti-inflammatory activity in human hepatoma HepG2 cells.
Butyric acid (BA) has been reported to induce anticancer effects on hepatocellular carcinoma (HCC) cells both in vitro and in vivo. However, its delivery and release in cancer tissues must be optimized. On the basis of these requirements, we prepared liposomes coated with chitosan and uncoated liposomes and both types were loaded with BA through a thin-film hydration method. The liposomes coated or uncoated with chitosan had a mean hydrodynamic size of 83.5 and 110.3 nm, respectively, with a homogeneous size distribution of the particles. For evaluation of the biological effects of the nanoformulations, the hepatoblastoma (HB) HepG2 cell line was utilized. BA-loaded liposomes coated with chitosan showed a considerable higher cytotoxicity than both uncoated liposomes and free BA, with IC50 values, after 72 h of incubation, of 7.5, 2.5 and 1.6 mM, respectively. Treatment of HepG2 cells for 5 h with the BA-loaded liposomes coated with chitosan at 5 mM lowered the extent of the increase in IL-8, IL-6, TNF-α and TGF-β expression of approximately 64, 58, 85 and 73.8%, respectively, when compared to the untreated cells. The BA-loaded liposomes coated with chitosan had marked capacity to be internalized in human HB cells showing an increased cytotoxic activity when compared with free BA and important anti-inflammatory effects by inhibiting production of cytokines with a central role in liver cell survival.


[Fear of having the antiepileptic drug withdrawn].
The decision to withdraw antiepileptic drugs is based mostly on a balanced view of the overall risk of seizure relapse and factors most likely to affect that risk. The aim of this study was to find out the patient and family's attitudes towards the fear of having antiepileptic drug withdrawn, after three years of seizure control. This research was carried out at the Institute for Child and Youth Health care of Vojvodina in Novi Sad. During the study, which lasted from 2003 to 2008, a face-to-face interview about fear of having the antiepileptic therapy withdrawn was done within the adolescent patient examination. The study population included 150 adolescent patients and 265 of their parents. In general, the adolescent patients were ready to accept a significantly higher risk of having recurrences after the antiepileptic drug withdrawal (p < 0.05) than their parents. None of their parents was ready to accept the risk 50% higher than the one in the general population. It is important to take into consideration the fear of both adolescent patients and their parents of having recurrences of epileptic seizures before opting for the antiepileptic drug withdrawal.


Unraveling Molecular Fingerprints of Catalytic Sulfur Poisoning at the Nanometer Scale with Near-Field Infrared Spectroscopy.
Fundamental understanding of catalytic deactivation phenomena such as sulfur poisoning occurring on metal/metal-oxide interfaces is essential for the development of high-performance heterogeneous catalysts with extended lifetimes. Unambiguous identification of catalytic poisoning species requires experimental methods simultaneously delivering accurate information regarding adsorption sites and adsorption geometries of adsorbates with nanometer-scale spatial resolution, as well as their detailed chemical structure and surface functional groups. However, to date, it has not been possible to study catalytic sulfur poisoning of metal/metal-oxide interfaces at the nanometer scale without sacrificing chemical definition. Here, we demonstrate that near-field nano-infrared spectroscopy can effectively identify the chemical nature, adsorption sites, and adsorption geometries of sulfur-based catalytic poisons on a Pd(nanodisk)/Al2O3 (thin-film) planar model catalyst surface at the nanometer scale. The current results reveal striking variations in the nature of sulfate species from one nanoparticle to another, vast alterations of sulfur poisoning on a single Pd nanoparticle as well as at the assortment of sulfate species at the active metal-metal-oxide support interfacial sites. These findings provide critical molecular-level insights crucial for the development of long-lifetime precious metal catalysts resistant toward deactivation by sulfur.


The molecular archaeology of a mitochondrial death effector: AIF in Drosophila.
Apoptosis-inducing factor (AIF) is a phylogenetically conserved redox-active flavoprotein that contributes to cell death and oxidative phosphorylation in Saccharomyces cerevisiae, Caenorhabditis elegans, mouse and humans. AIF has been characterized as a caspase-independent death effector that is activated by its translocation from mitochondria to the cytosol and nucleus. Here, we report the molecular characterization of AIF in Drosophila melanogaster, a species in which most cell deaths occur in a caspase-dependent manner. Interestingly, knockout of zygotic D. melanogaster AIF (DmAIF) expression using gene targeting resulted in decreased embryonic cell death and the persistence of differentiated neuronal cells at late embryonic stages. Although knockout embryos hatch, they undergo growth arrest at early larval stages, accompanied by mitochondrial respiratory dysfunction. Transgenic expression of DmAIF misdirected to the extramitochondrial compartment (DeltaN-DmAIF), but not wild-type DmAIF, triggered ectopic caspase activation and cell death. DeltaN-DmAIF-induced death was not blocked by removal of caspase activator Dark or transgenic expression of baculoviral caspase inhibitor p35, but was partially inhibited by Diap1 overexpression. Knockdown studies revealed that DeltaN-DmAIF interacts genetically with the redox protein thioredoxin-2. In conclusion, we show that Drosophila AIF is a mitochondrial effector of cell death that plays roles in developmentally regulated cell death and normal mitochondrial function.


Evaluation of homogeneity of binary blends of poly(3-hydroxybutyrate) and poly(L-lactic acid) studied by near infrared chemical imaging (NIRCI).
The homogeneity of blends of poly((R)-3-hydroxybutyrate) (PHB) and poly(L-lactic acid) (PLLA) was evaluated by the near infrared chemical imaging (NIRCI) technique. NIRCI can nondestructively investigate a sample over a wide field of view within a few minutes to acquire a large number of spatially resolved NIR spectral data. NIRCI may be combined with multivariate analysis not only for qualitative analysis but also for statistically based quantitative analysis. The score images derived from the partial least squares regression (PLSR) analysis directly show that PHB/PLLA blends are highly homogeneous. The standard deviations (STD) of the histograms, indicating the distribution of the score values, show small values for the blends. These results qualitatively and quantitatively show the high level of homogeneity of PHB/PLLA blends. The predictions of the spatially averaged concentrations of the blend components obtained from PLSR results show values similar to the actual contents for the blends. The small errors of the predictions are also explained by STD values.


Impact of Cumulative Smoking Exposure on Subclinical Degenerative Aortic Valve Disease in Apparently Healthy Male Workers.
Little is known regarding the impact of cumulative smoking exposure and smoking cessation on subclinical degenerative aortic valve (DAV) disease. We examined associations of smoking status, cigarette-years of smoking, and years since quitting smoking with subclinical DAV disease defined by echocardiography. Of 756 apparently healthy male workers, 154 had DAV including 63 with DAV with ≥2 leaflets calcification (DAV ≥2 cal). Compared with never smokers, ever smokers had higher risk of DAV and DAV ≥2 cal; odds ratios (95% CI) were 2.883 (1.800-4.618) (p<0.001) and 5.281 (2.297-12.138) (p<0.001), respectively. Both current and former smokers had dose-dependent relationships of cigarette-years on DAV and DAV ≥2 cal (P for trend, <0.001 for both DAV, in both smokers). In current smokers with >400-≤800 and with >800 cigarette-years, odds ratios (95%CIs) were 3.201 (1.690-6.063) (p<0.001) and 5.326 (2.800-10.053) (p<0.001) for DAV, 7.460 (2.828-19.680) (p<0.001) and 8.397 (3.146-22.414) (p<0.001) for DAV ≥2 cal, respectively. In former smokers with >800 cigarette-years, odds ratios (95%CI) were 3.780 (1.970-7.254) (p<0.001) for DAV, 10.035 (3.801-26.496) (p<0.001) for DAV ≥2 cal. Compared with current smokers, former smokers with quitting smoking >10 years had significantly lower risk of DAV and DAV ≥2 cal. In apparently healthy male workers, DAV disease was strongly associated with smoking. Cumulative smoking exposure was associated with dose-dependent relationship on subclinical DAV disease both in current smokers and former smokers.


Surface imprinted bacterial cellulose nanofibers for hemoglobin purification.
There is a significant need for the development of the novel adsorbents in the field of protein purification. In this study, thin hemoglobin imprinted film (MIP) was fabricated onto the bacterial cellulose nanofibers' (BCNFs) by surface imprinting method using metal ion coordination interactions with N-methacryloyl-(L)-histidinemethylester (MAH) and copper ions. The hemoglobin surface imprinted bacterial cellulose nanofibers (MIP-BCNFs) was applied to selective recognition of hemoglobin and purification from hemolysate. The characterization of the MIP-BCNFs was carried out by the Fourier Transformed Infrared Spectroscopy (FT-IR), Scanning Electron Microscopy (SEM), micro-Computerized Tomography (μCT), atomic force microscopy (AFM) and surface area measurements. The adsorption experiments of hemoglobin onto the MIP-BCNFs and NIP-BCNFs from aqueous hemoglobin solutions were investigated in a batch system. The results showed that MIP-BCNFs are promising materials for purification of hemoglobin with high adsorption capacity, significant selectivity and reusability.


Racial/Ethnic Differences in Mental Health, Substance Use, and Bullying Victimization Among Self-Identified Bisexual High School-Aged Youth.
Purpose: Sexual minority youth are at increased risk for mental health problems and substance use, and accumulating evidence indicates that bisexual youth are at greatest risk. However, bisexual youth are not a homogenous group and scholars have called for greater attention to the intersections of multiple marginalized identities. As such, we examined racial/ethnic differences in mental health (sadness/hopelessness and suicidal ideation), substance use (cigarette use, binge drinking, marijuana use, and other illicit drug use), and bullying (in-person and electronic) among self-identified bisexual high school-aged youth (overall and by sex). Method: Data from the local versions of the Youth Risk Behavior Survey were pooled across jurisdictions and years (2011-2015), resulting in an analytic sample of 18,515 bisexual youth who were racially/ethnically diverse. Results: Black and Hispanic bisexual youth were less likely to report in-person and electronic bullying than White bisexual youth. In addition, Black bisexual youth were less likely to report sadness/hopelessness and suicidal ideation than White, Hispanic, and Other race/ethnicity bisexual youth. Black bisexual female youth were also less likely to report cigarette use, binge drinking, and other illicit drug use than White bisexual female youth. In contrast to most of our findings, Black bisexual youth were more likely to report marijuana use than White bisexual youth. Most of the significant racial/ethnic differences in mental health and substance use remained significant after controlling for bullying. Conclusion: These findings highlight the heterogeneity of bisexual youth and the need to consider multiple marginalized identities to understand the health disparities affecting this diverse population.


Aspiration biopsy smear pattern as a predictor of biologic behavior in adenocarcinoma of the breast.
Fine needle aspiration cytology (FNAC) is increasingly used for the initial diagnosis of breast carcinoma, yet few studies have investigated the prognostic importance of cytologic features seen in breast FNAC. We studied the relationship between prognosis and smear morphology in 26 patients with breast carcinoma for whom information from long-term follow-up was available. Specifically, we attempted to correlate nuclear grade and smear pattern with hormonal receptor status, lymph node status, breast recurrence and distant metastasis. In an additional five patients (on whom insufficient follow-up information was available), smear pattern and nuclear grade were correlated with hormonal status. Smears were classified into three patterns: typical (sheets and clusters), individual cell predominant and cluster predominant. The individual cell predominant pattern was associated with an increased incidence of distant metastasis when compared with the other two smear patterns (P less than .05). No statistically significant association was observed between smear pattern and the other factors studied. Nuclear grade 2 (of 3) was also associated with an increased incidence of distant metastasis. However, most grade 2 tumors (10/12) were of the individual cell predominant pattern. Smear pattern classification, along with DNA ploidy analysis and hormone receptor analysis of FNAC-derived material, may yield valuable prognostic information.


Acanthamoeba keratitis therapy: time to cure and visual outcome analysis for different antiamoebic therapies in 227 cases.
To test the hypothesis that Acanthamoeba keratitis (AK) outcomes differ for different topical antiamoebic therapies (AAT) and to provide the detailed patient outcome data. A retrospective cohort study of 227 patients developing AK between 25 July 1991 and 10 August 2012. Inclusion criteria required a complete record of AAT treatment for both the primary outcome of a medical cure rate at 12 months and the secondary outcome of Snellen visual acuity ≤6/24 and/or surgical intervention. Analysis used multivariable regression to control for differences in baseline disease characteristics for both primary and secondary outcomes with unadjusted analyses for other outcomes. Subjects were categorised for analysis both by the AAT used at baseline and also by mutually exclusive AAT (patients exposed to all the drugs in each group, and no others, for some period). AAT categories were PHMB monotherapy, PHMB+diamidine, PHMB+chlorhexidine+diamidine, diamidine monotherapy and other AAT. Analysis by baseline AAT showed no notable difference between treatments for both a medical cure at 12 months in 60.79% (138/227) or for a poor outcome in 49.34% (112/227). When AATs were analysed by mutually exclusive groups, PHMB monotherapy provided the best outcomes. These findings are subject to bias requiring careful interpretation. Overall cure rates for the 214 subjects with resolved outcomes were 94.27% (214/227), median time to cure 5 months (IQR 3.25-9.00 months) and range 1-26.24 months. PHMB 0.02% monotherapy for the initial treatment of AK is as effective as biguanide+diamidine combination therapy. Chlorhexidine monotherapy was too infrequent for comparison. The outcome data are the most detailed available.


Changes in biocrust cover drive carbon cycle responses to climate change in drylands.
Dryland ecosystems account for ca. 27% of global soil organic carbon (C) reserves, yet it is largely unknown how climate change will impact C cycling and storage in these areas. In drylands, soil C concentrates at the surface, making it particularly sensitive to the activity of organisms inhabiting the soil uppermost levels, such as communities dominated by lichens, mosses, bacteria and fungi (biocrusts). We conducted a full factorial warming and rainfall exclusion experiment at two semiarid sites in Spain to show how an average increase of air temperature of 2-3 °C promoted a drastic reduction in biocrust cover (ca. 44% in 4 years). Warming significantly increased soil CO2 efflux, and reduced soil net CO2 uptake, in biocrust-dominated microsites. Losses of biocrust cover with warming through time were paralleled by increases in recalcitrant C sources, such as aromatic compounds, and in the abundance of fungi relative to bacteria. The dramatic reduction in biocrust cover with warming will lessen the capacity of drylands to sequester atmospheric CO2 . This decrease may act synergistically with other warming-induced effects, such as the increase in soil CO2 efflux and the changes in microbial communities to alter C cycling in drylands, and to reduce soil C stocks in the mid to long term.


Molecular Targeted Therapies Using Botanicals for Prostate Cancer Chemoprevention.
In spite of the large number of botanicals demonstrating promise as potential cancer chemopreventive agents, most have failed to prove effectiveness in clinical trials. Critical requirements for moving botanical agents to recommendation for clinical use include adopting a systematic, molecular-target based approach and utilizing the same ethical and rigorous methods that are used to evaluate other pharmacological agents. Preliminary data on a mechanistic rationale for chemoprevention activity as observed from epidemiological, in vitro and preclinical studies, phase I data of safety in suitable cohorts, duration of intervention based on time to progression of pre-neoplastic disease to cancer and using a valid panel of biomarkers representing the hypothesized carcinogenesis pathway for measuring efficacy must inform the design of clinical trials. Botanicals have been shown to influence multiple biochemical and molecular cascades that inhibit mutagenesis, proliferation, induce apoptosis, suppress the formation and growth of human cancers, thus modulating several hallmarks of carcinogenesis. These agents appear promising in their potential to make a dramatic impact in cancer prevention and treatment, with a significantly superior safety profile than most agents evaluated to date. The goal of this paper is to provide models of translational research based on the current evidence of promising botanicals with a specific focus on targeted therapies for PCa chemoprevention.


Multiple paternity and population genetic structure in natural populations of the poeciliid fish, Heterandria formosa.
Heterandria formosa Agassiz, exhibits internal fertilization, internal brooding of embryos, sperm storage and an extreme level of superfetation. In this study we used microsatellite markers to examine variation among seven populations that exhibited significant variance in their histories of population density. We found that the populations were genetically distinct and that the heterozygosity increased as population density increased. We also examined paternity in three of those populations and found that the number of sires per female and the number of sires per brood increased with population density. Overall, the rates of multiple paternity are quite low relative to other species. The correlations with population density suggest that contact rates play a critical role in the breeding system in this species but the low rates of multiple paternity suggest that females may exert control over fertilization of their ova.


Measurable outcomes of instructions to modify normal parent-child verbal interactions: implications for indirect stuttering therapy.
This study was a preliminary analysis of the differential effects of instructions to slow maternal speech rate, or to slow and simplify maternal speech when conversing with normally fluent children. Twenty mothers and their normally fluent children participated. Instructions to slow maternal rate resulted in maternal speech simplification as a by-product. No significant differences were found between the effects of the two sets of instructions. Furthermore, children's speech rate and language complexity did not parallel maternal adjustments. These last findings are problematic for some of the premises that underlie the use of parental counseling as a component of indirect therapy for stuttering.


Organ damage in Atlantic herring larvae as a result of ocean acidification.
The dissolution of anthropogenically emitted excess carbon dioxide lowers the pH of the world's ocean water. The larvae of mass spawning marine fishes may be particularly vulnerable to such ocean acidification (OA), yet the generality of earlier results is unclear. Here we show the detrimental effects of OA on the development of a commercially important fish species, the Atlantic herring (Clupea harengus). Larvae were reared at three levels of CO2: today (0.0385 kPa), end of next century (0.183 kPa), and a coastal upwelling scenario (0.426 kPa), under near-natural conditions in large outdoor tanks. Exposure to elevated CO2 levels resulted in stunted growth and development, decreased condition, and severe tissue damage in many organs, with the degree of damage increasing with CO2 concentration. This complements earlier studies of OA on Atlantic cod larvae that revealed similar organ damage but at increased growth rates and no effect on condition.


[Characteristics of rehabilitation in patients with rheumatic diseases].
The paper deals with problems of rehabilitation of patients with rheumatoid arthritis, osteoarthrosis and systemic lupus erythematosus. These problems are rather topical in view of high prevalence of rheumatic diseases, their severe course and a high rate of invalidization at the most productive age. Relevant multifactorial therapeutic measures should be staged, aimed at lowering and stabilization of the disease activity. The role of updated medication, joint surgery, diet therapy, etc. is shown.


Differential localization of colonic H(+)-K(+)-ATPase isoforms in surface and crypt cells.
Two distinct colonic H(+)-K(+)-adenosinetriphosphatase (H(+)-K(+)-ATPase) isoforms can be identified in part on the basis of their sensitivity to ouabain. The colonic H(+)-K(+)-ATPase alpha-subunit (HKc alpha) was recently cloned, and its message and protein are present in surface (and the upper 20% of crypt) cells in the rat distal colon. These studies were performed to establish the spatial distribution of the ouabain-sensitive and ouabain-insensitive components of both H(+)-K(+)-ATPase activity in apical membranes prepared from surface and crypt cells and K(+)-dependent intracellular pH (pHi) recovery from an acid load both in isolated perfused colonic crypts and in surface epithelial cells. Whereas H(+)-K(+)-ATPase activity in apical membranes from surface cells was 46% ouabain sensitive, its activity in crypt apical membranes was 96% ouabain sensitive. Similarly, K(+)-dependent pHi recovery in isolated crypts was completely ouabain sensitive, whereas in surface cells K(+)-dependent pHi recovery was insensitive to ouabain. These studies provide compelling evidence that HKc alpha encodes the colonic ouabain-insensitive H(+)-K(+)-ATPase and that a colonic ouabain-sensitive H(+)-K(+)-ATPase isoform is present in colonic crypts and remains to be cloned and identified.


Brucellosis and uveitis.
A 30-year-old-lady, (Case 1) was found to have brucella uveitis in her left eye. Serum agglutination (SAT) and SAT/Coombs titers were positive at 1:320 and 1:160, respectively. In Case 2, a 12-year-old-girl diagnosis of left brucellosis uveitis was made. Both SAT and SAT/Coombs titers were positive at 1:1280. This report confirms that Brucellae organisms as a cause of uveitis.


Isothiocyanates: An Overview of Their Antimicrobial Activity against Human Infections.
The use of plant-derived products as antimicrobial agents has been investigated in depth. Isothiocyanates (ITCs) are bioactive products resulting from enzymatic hydrolysis of glucosinolates (GLs), the most abundant secondary metabolites in the botanical order Brassicales. Although the antimicrobial activity of ITCs against foodborne and plant pathogens has been well documented, little is known about their antimicrobial properties against human pathogens. This review collects studies that focus on this topic. Particular focus will be put on ITCs' antimicrobial properties and their mechanism of action against human pathogens for which the current therapeutic solutions are deficient and therefore of prime importance for public health. Our purpose was the evaluation of the potential use of ITCs to replace or support the common antibiotics. Even though ITCs appear to be effective against the most important human pathogens, including bacteria with resistant phenotypes, the majority of the studies did not show comparable results and thus it is very difficult to compare the antimicrobial activity of the different ITCs. For this reason, a standard method should be used and further studies are needed.


Spectroscopic and calorimetric evaluation of chemically induced protein denaturation in HuH-7 liver cancer cells and impact on cell survival.
Solid tumors such as hepatocellular carcinoma are very often not amenable to chemotherapy and radiotherapy. Local ablation methods, including chemical ablation with absolute ethanol, are therefore an option for treatment but lack of information about the mechanism of devitalization leading to cell death is a hindrance to further adoption. Systemic toxicity also has limited the amount of ethanol that can be used in a single treatment session. Therefore we evaluated the mechanism of urea, a denaturant with little or no systemic toxicity, for potential use in chemical ablation. In this study we report on the use of three methods to analyze the effects in cell culture with a view towards eventual clinical application. Human hepatoma HuH-7 cells were analyzed at several time points after treatment using FTIR, DSC, and Raman microspectroscopy based on MTT and PI-exclusion viability assays. Time course fractional denaturation data plotted against viability show that a 50% viability drop occurs after only a 10-20% drop in overall protein denaturation. Other methods of cell death such as apoptosis may also be operative, but this result implies that protein denaturation is one of the major mechanisms of cell death. This is in line with what has been previously suggested for purely thermal methods, and opens the way to mechanism-based improvements in chemical ablation of solid tumors.


Elaboration, activity and stability of silica-based nitroaromatic sensors.
Functionalized silica-based thin films, modified with hydrophobic groups, were synthesized and used as sensors for nitroaromatic compound (NAC) specific detection. Their performance and behavior, in terms of stability, ageing and regeneration, have been fully characterized by combining chemical characterization techniques and electron microscopy. NAC was efficiently and specifically detected using these silica-based sensors, but showed a great degradation in the presence of humidity. Moreover, the sensor sensitivity seriously decreases with storage time. Methyl- and phenyl-functionalization helped to overcome this humidity sensitivity. Surface characterization enabled us to establish a direct correlation between the appearance, and increasing amount, of adsorbed carbonyl-containing species, and sensor efficiency. This contamination, appearing after only one month, was particularly important when sensors were stored in plastic containers. Rinsing with cyclohexane enables us to recover part of the sensor performance but does not yield a complete regeneration of the sensors. This work led us to the definition of optimized elaboration and storage conditions for nitroaromatic sensors.


Lessons learned from studies of the natural history of diabetic nephropathy in young type 1 diabetic patients.
Diabetic Nephropathy (DN) remains the leading cause of end stage renal disease (ESRD) in the Western world, responsible for nearly half of all new ESRD cases in the USA (1). DN develops in 20-25% of patients with type 1 diabetes (T1DM) (2) and, although risk of DN is clearly related to glycemic control (3,4), other variables including genetic propensity (5) are needed to explain why only a minority T1 DN patients progress to ESRD. The clinical manifestations of DN including increasing levels of urinary albumin excretion (AER), rising blood pressure (BP) and falling glomerular filtration rates (GFR) are closely related to renal structural abnormalities of DN (5,6). These glomerular, tubular, vascular and interstitial lesions are strongly correlated with these functional abnormalities especially when non-linear analysis models are used (6,7). This is because DN's natural history is one of clinical silence for years to decades during which time serious underlying renal lesions may be developing. Once the clinical manifestations, including the development of persistent microalbuminuria [(MA); (AER 20-200 microg/min)] are present, the structural injury is often far advanced (8). Moreover the nature of the renal lesions changes following the development of overt proteinuria so that the further decline in GFR is now associated with focal and global glomerular sclerosis and tubulo-interstitial injury which probably accelerates the GFR decline towards ESRD (7). Since interventions at these late stages of disease may only slow but not completely arrest the inexorable progression towards renal failure (9), understanding early natural history becomes important. Since DN structurally and functionally is a progressive disease; it is reasonable to presume that patients that either do not develop the earlier lesions of DN or develop them very slowly will not progress within their lifetime to stages of advanced renal structural injury and ESRD. We therefore considered it important to understand the early natural history of diabetic nephropathy and formed the International Diabetic Nephropathy Study Group (IDNSG) in order to investigate the early stages of DN in young T1DM volunteers. The design of the Natural History Study (NHS) (9) has been reported. The IDNSG participating institutions included 3 university centers (McGill University, Montreal, Canada with affiliations with the University of Sherbrooke, Sherbrooke, Canada, the Ottawa Civic Hospital, and the Childrens Hospital of Eastern Ontario; the Department of Pediatrics, University of Minnesota Medical School with affiliations at St Paul Children's Hospital and the International Diabetes Center in Minneapolis; the Robert Debré Höpital in Paris with affiliations with Höpital Saint Louis). The data coordinating center for the NHS was in the Department of Epidemiology and Biostatistics at McGill University and light microscopy readings were carried out at INSERM Unité 192 at the Höpital Necker-Enfants Malades in Paris. Patients could be included if they had type 1 diabetes for 2-20 years, had onset of diabetes before age 31, had AER less than 100 mug/min and GFR > or = 90 ml/min/ 1.73m2 (9). Patients also had to be normotensive for their age and sex and have no other significant renal or systemic disease. Quarterly studies included measurements of blood pressure, (BP), urinary albumin excretion rate (AER), hemoglobin A1C (HbA1C), GFR, and renal plasma flow (RPF). Renal biopsies were performed at baseline and after 5 years in the study. The primary goal of the study was to determine the clinical predictors of the baseline biopsy and baseline clinical and renal structural predictors of the changes between the baseline and the 5 year biopsy. The longitudinal structural studies are still in analysis and this paper will mainly review the cross sectional studies that have been completed to date.


Role of childhood adversities and environmental sensitivity in the development of post-traumatic stress disorder in war-exposed Syrian refugee children and adolescents.
Increased post-traumatic stress disorder (PTSD) rates have been documented in children exposed to war. However, the contribution of childhood adversities and environmental sensitivity to children's responses to adversities and trauma are still far from settled.AimsTo evaluate the relative roles of war, childhood adversities and sensitivity in the genesis of PTSD. Data on childhood adversities and sensitivity was collected from 549 Syrian refugee children in Lebanon. PTSD symptoms were assessed using the PTSD Reaction Index. Although childhood adversities, war events and sensitivity were all significantly related to PTSD in bivariate analyses, multivariate analyses showed that childhood adversities were the most important variable in predicting PTSD. The effect of war on PTSD was found to be dependent on the interplay between childhood adversities and sensitivity, and was most prominent in highly sensitive children with lower levels of adversities; in sensitive children experiencing high levels of adversities, the effects of war exposure on PTSD were less pronounced. When considering the effects of war on PTSD in refugee children, it is important to take account of the presence of other adversities as well as of children's sensitivity. Sensitive children may be more vulnerable to the negative effects of war exposure, but only in contexts that are characterised by low childhood adversities.Declaration of interestNone.


Formation of Ge nanocrystals in Al2O3 matrix.
Ge nanocrystals were formed in Al2O3 matrix by implantation of Ge ions into sapphire (alpha-Al2O3) substrates and subsequent annealing. Diagnostic techniques, Raman spectroscopy, XRD, TEM, EDS, and SAED were employed to monitor and study formation of Ge nanocrystals and their evolution during heat treatments. TEM and EDS analysis revealed the diffusion of Ge ions into the substrate during annealing process. While Ge nanocrystals with mean sizes of 15 nm were observed in the heavily implanted region small nanocrystals with mean sizes of 4 nm were identified underneath this region. Some grains of transition aluminas were formed in the implanted region which was amorphized during the implantation. Extensive stress between the transition aluminas and sapphire matrices and its effects on the matrix were detected. The effect of stress on the Raman and XRD spectra of Ge nanocrystals was discussed.


Transganglionic degeneration of capsaicin-sensitive C-fiber primary afferent terminals.
Local treatment of rat peripheral nerves with capsaicin induces permanent impairment of afferent C-fiber functions and changes in the response properties of spinal dorsal horn neurons. In this study a new experimental approach, the "capsaicin gap" technique, has been utilized in an attempt to unravel pathomorphological alterations which commence in the domain of primary sensory neurons as a consequence of perineural treatment with capsaicin. The technique relies on the facts that peripheral nerves in the spinal dorsal horn are represented in a strict somatotopic manner, and on the observation that in the adult rat systemic injection of appropriate doses of capsaicin results in a selective degeneration of primary afferent fibers within Rexed's laminae I and II of the spinal cord. We have assumed that, if treatment of a peripheral nerve with capsaicin results in an irreversible destruction of primary afferent terminals related to that nerve, then the corresponding spinal projection areas will be free of degeneration following a subsequent systemic injection of capsaicin. The topographical distribution of degenerating axon terminals, induced by a systemic injection of capsaicin, was studied within the lumbar spinal cord of rats whose sciatic nerves had been treated with capsaicin or its vehicle one to three months previously. In the dorsal horn ipsilateral to the vehicle-treated nerve, axon terminal degeneration was found in the entire mediolateral extent of Rexed's laminae I and II. In contrast, in the dorsal horn ipsilateral to the capsaicin-treated nerve, virtually no degeneration was observed in areas which correspond to the projection territories of sciatic afferents. It is concluded that the central terminals of capsaicin-sensitive sciatic afferents underwent transganglionic degeneration as a result of prior perineural treatment with capsaicin, and a subsequent systemic injection of this neurotoxin therefore failed to cause axon terminal degeneration in somatotopic areas served by the treated nerve. Comparative quantitative morphometric analysis of cell populations of dorsal root ganglia related to capsaicin- or vehicle-treated nerves disclosed (1) a marked reduction in the proportion of small-sized neurons, (2) a fall of about 80% in the percentage of neurons which undergo degeneration after the systemic injection of capsaicin, and (3) a marked decrease in the total number of neurons in ganglia ipsilateral to the capsaicin-treated nerves. Quantitative electron microscopic studies on saphenous nerves treated perineurally with capsaicin revealed a 32% reduction in the number of unmyelinated axons as compared with the controls, whereas the number of myelinated fibers was unchanged.(ABSTRACT TRUNCATED AT 400 WORDS)


Promoting vascular repair in the retina: can stem/progenitor cells help?
Since its first epidemic in the 1940s, retinopathy of prematurity (ROP) has been a challenging illness in neonatology. Higher than physiological oxygen levels impede the development of the immature retinal neuropil and vasculature. Current treatment regimens include cryotherapy, laser photocoagulation, and anti-VEGF agents. Unfortunately, none of these approaches can rescue the normal retinal vasculature, and each has significant safety concerns. The limitations of these approaches have led to new efforts to understand the pathological characteristics in each phase of ROP and to find a safer and more effective therapeutic approach. In the era of stem cell biology and with the need for new treatments for ROP, this review discusses the possible future use of unique populations of proangiogenic cells for therapeutic revascularization of the preterm retina.


Functional and non-functional nerve-smooth muscle transmission in the renal arteries of the newborn and adult rabbit and guinea-pig.
Isolated renal arteries from newborn and adult guinea-pigs failed to respond to electrical stimulation of the intramural nerves, using parameters identical to those which elicit large neurogenic contractions in renal arteries from newborn and adult rabbits. The threshold stimulation frequency was lower and responses were greater in adult compared with neonatal rabbits. Comparisons of noradrenaline EC50 values showed that guinea-pig renal arteries are significantly less sensitive to noradrenaline that rabbit vessels. Quantitative fluorescence histochemistry of the perivascular adrenergic nerves in renal arteries revealed that the density of innervation gradually declines in the guinea-pig but substantially increases in the rabbit during postnatal development. The lack of neurogenic response of the guinea-pig renal artery, despite the existence of a varicose, adrenergic nerve plexus, is discussed.


Determination of critical community size from an HIV/AIDS model.
After an epidemic outbreak, the infection persists in a community long enough to engulf the entire susceptible population. Local extinction of the disease could be possible if the susceptible population gets depleted. In large communities, the tendency of eventual damp down of recurrent epidemics is balanced by random variability. But, in small communities, the infection would die out when the number of susceptible falls below a certain threshold. Critical community size (CCS) is considered to be the mentioned threshold, at which the infection is as likely as not to die out after a major epidemic for small communities unless reintroduced from outside. The determination of CCS could aid in devising systematic control strategies to eradicate the infectious disease from small communities. In this article, we have come up with a simplified computation based approach to deduce the CCS of HIV disease dynamics. We consider a deterministic HIV model proposed by Silva and Torres, and following Nåsell, introduce stochasticity in the model through time-varying population sizes of different compartments. Besides, Metcalf's group observed that the relative risk of extinction of some infections on islands is almost double that in the mainlands i.e. infections cease to exist at a significantly higher rate in islands compared to the mainlands. They attributed this phenomenon to the greater recolonization in the mainlands. Interestingly, the application of our method on demographic facts and figures of countries in the AIDS belt of Africa led us to expect that existing control measures and isolated locations would assist in temporary eradication of HIV infection much faster. For example, our method suggests that through systematic control strategies, after 7.36 years HIV epidemics will temporarily be eradicated from different communes of island nation Madagascar, where the population size falls below its CCS value, unless the disease is reintroduced from outside.


Magnetic retraction of bowel by intraluminal injectable cyanoacrylate-based magnetic glue.
Magnetic retraction offers advantages over physical retraction by graspers because of reduced tissue trauma. The objectives of this study are to investigate a novel method of magnetisation of bowel segments by intraluminal injection of magnetic glue and to demonstrate the feasibility of magnetic retraction of bowel with sufficient force during minimal access surgery. Following an initial materials characterisation study, selected microparticles of stainless steel (SS410- μ Ps) were mixed with chosen cyanoacrylate glue (Loctite 4014). During intraluminal injection of the magnetic glue using ex vivo porcine colonic segments, a magnetic probe placed at the injected site ensured that the SS410-μPs aggregated during glue polymerisation to form an intraluminal mucosally adherent coagulum. The magnetised colonic segments were retracted by magnetic probes (5 and 10 mm) placed external to the bowel wall. A tensiometer was used to record the retraction force. With an injected volume of 2 mL in a particle concentration of 1 g/mL, this technique produced maximal magnetic retraction forces of 2.24 ± 0.23 N and 5.11 ± 0.34 N (n = 20), with use of 5 and 10 mm probes, respectively. The results indicate that the formation of an intraluminal coagulum based on SS410- μPs and Loctite 4014 produces sufficient magnetic retraction for bowel retraction.


Regeneration of spermatogenesis and production of functional sperm by grafting of testicular cell aggregates in Zebrafish (Danio rerio).
The self-renewal and differentiation of spermatogonial stem cells (SSCs) is essential for the continuous production of sperm throughout life in male vertebrates. The development of a functional assay to analyze these properties in isolated SSCs remains necessary. In our current study, we have developed a transplantation method for testicular cell aggregates in zebrafish (Danio rerio) in which allogeneic SSCs can undergo self-renewal and differentiation. The immature testes from juveniles are dissociated, aggregated by cultivation, and then transplanted under the abdominal skin of the recipient fish. The grafted aggregates reconstitute the appropriate testicular structures, including the lobule structure, consisting of basement membrane and interstitial steroid-producing cells on the outside, and the cysts, which comprise germ cell clusters and surrounding Sertoli cells. Bromodeoxyuridine incorporation analysis indicated that continuous spermatogenesis is maintained for at least 6 mo in the reconstituted testis. Moreover, when the sperm generated from the aggregates at 3 mo postgrafting were used for artificial insemination, fertilized eggs were obtained that developed sexually mature fish. These results suggest that self-renewal of SSCs takes place in reconstituted testes under the abdominal skin and that their differentiating progeny can develop into functional sperm. Furthermore, allogeneic spermatogonia were also found to proliferate and differentiate into sperm in these grafts. Our method of grafting testicular cell aggregates should thus prove useful not only analyzing the stem cell ability of an individual SSC but also for the production of progeny from cultured SSCs or SSCs of sterile mutants with somatic cell defects.


The Impact of the COVID-19 Pandemic in Adolescents with Asthma.
The coronavirus disease 2019 (COVID-19) pandemic is affecting people at any age and there is limited information about the effect of the COVID-19 pandemic on quality of life (QoL) in adolescents with asthma. In the present study, it was aimed to assess the attitudes of adolescents with asthma toward the COVID-19 pandemic and determine the effects of the pandemic on their QoL. In total, 125 adolescents with asthma and 98 healthy adolescents participated in the present study. The questionnaire form consisted of three parts. In the first part, all the participants were asked whether they complied with the protective measures against COVID-19. The second part included questions for measuring the participants' level of concern about COVID-19, while the third part consisted of EUROHIS-QOL 8. The patient and control groups were similar in terms of the female/male ratio (55/70 and 48/50, respectively) and mean participant age (14.6 ± 2 and 15.1 ± 1.65 years, respectively) (P = 0.459 and P = 0.062, respectively). The prevalence of COVID-19 in the patients (n = 2, 1.6%) was lower than that in the controls (n = 6, 6.1%); however, the difference was not statistically significant (P = 0.142). The total EUROHIS-QOL score was significantly lower in the patients (31.2 ± 6.7) than in the controls (33.7 ± 4.4) (P < 0.001). The total QoL scores of asthmatic adolescents without other allergic disease (31.4 ± 6.7) was also lower than those of the controls (33.7 ± 4.4) (P = 0.009). Treatment disruption was significantly more common in patients who received subcutaneous immunotherapy (n = 20, 48.8%) than in those who did not (n = 8, 9.5%) (P < 0.001). Moreover, the patients had lower EUROHIS-QOL scores in the overall QoL, general health, finance, and home domains. Our results indicate that the mean QoL score of asthmatic adolescents during COVID-19 pandemic is lower than in the healthy population. Disruption in their treatment was most common in patients with asthma who were receiving subcutaneous immunotherapy.


Mutant alleles at the locus elav in Drosophila melanogaster lead to nervous system defects. A developmental-genetic analysis.
We report a developmental and genetic analysis of the X-linked vital locus l(1)EC7 in Drosophila melanogaster. The locus maps in the salivary band region 1B4-5 to 1B8-9, a part of the X chromosome previously shown to be essential for normal neural development. Certain mutant alleles at the locus can cause embryonic lethality, indicating that the function provided by the gene is essential during embryogenesis. A developmental analysis of gynandromorphic genetic mosaics shows that: (1) the gene function is autonomously essential in the eye; (2) the gene function is essential for normal development of the optic lobes; and (3) the gene function is not necessary in most major imaginal-disc cell derivatives with the exception of the eye disc. Conclusions from the developmental analysis of a temperature sensitive allele are consistent with those from the mosaic analysis. The embryonic lethality caused by the mutant alleles and abnormalities observed in the genetic mosaics have led us to rename the locus l(1)EC7 to elav (embryonic lethal, abnormal visual system).


Measles antibodies in cord blood in Portugal: Possible consequences for the recommended age of vaccination.
The optimum age to give the first dose of measles vaccine must balance the risks of disease and vaccine failure. Both are influenced by the levels of transplacentally acquired maternal antibodies. This study was conducted in the Obstetric service of Portuguese hospital, in 2012-2013. Mothers were recruited after informed consent. Measles IgG was measured in 206 cord sera, using a commercial immunoassay. Geometric mean concentrations (and 95% CI) were 1849mIU/ml (1196-2857) and 790mIU/ml (618-1008) in cord sera of newborns from unvaccinated and vaccinated mothers respectively. Maternal age and vaccination status were both associated with the concentration in cord sera, but maternal age was the major predictor. The likely explanation is the same already mentioned in other studies: as a vaccination program progresses, vaccination coverage increases as measles incidence decreases. That results newborns from younger vaccinated mothers having less measles antibodies while the older mothers are more likely to have been infected with the wild virus. As the proportion of vaccinated mothers increase, developed countries tend to anticipate the recommended age of the first dose to 12 months of age. Models using hypothetical measles antibody decay rates in infancy were explored. Anticipating the first dose of MMR1 in Portugal to the age of 12 months might have not been the best decision but results were not conclusive, and arguments supporting or not the anticipation were discussed.


Satellite-like particle of tobacco ringspot virus that resembles tobacco ringspot virus.
A satellite-like nucleoprotein serologically indistinguishable from multicomponent tobacco ringspot virus, also resembles it in size, shape, and electrophoretic mobility. Although the protein shell of the nucleoprotein is similar to, if not identical to that of tobacco ringspot virus, each ahell of the nucleoprotein conntains many small strands of nucleic acid, which replicate only in mixed infections with the virus.


Carrot phytoalexin alters the membrane permeability of Candida albicans and multilamellar liposomes.
The biochemical basis for the antimicrobial effect of the carrot phytoalexin 6-methoxymellein (6-MM) was examined. At fungistatic concentrations 6-MM retarded the ability of Candida albicans to incorporate radioactive thymidine, uridine and leucine into biopolymers. When C. albicans was incubated with 6-MM, 260-nm-absorbing materials and 3H-labelled compounds leaked from the cells. The inhibitory effects of 6-MM on cell growth and membrane functions were, however, reduced as the concentration of divalent metal cations added to the medium was increased. 6-MM interacted with multilamellar liposomes constituted from phosphatidylcholine, cholesterol and dicetyl phosphate, or from phosphatidylcholine only, resulting in the release of glucose trapped in these liposomes. These results suggest that 6-MM exerts its toxic effects on susceptible cells as a result of its interaction with their membranes and disturbance of membrane-associated functions.


Multiple, thin-walled cysts are one of the HRCT features of airspace enlargement with fibrosis.
Airspace enlargement with fibrosis (AEF) has been identified pathologically as a smoking related change. We sought to identify the HRCT findings of AEF and search for distinguishing features from honeycombing. 50 patients (47 males; mean age 69) were evaluated. All had undergone lobectomy for lung cancer and had confirmed AEF and/or usual interstitial pneumonia (UIP) by pathological evaluation. HRCT findings were first evaluated preresection for resected lobes, and then correlated with the subsequent pathological findings in the resection specimens. Three groups were devised: one with AEF alone to determine the HRCT findings of AEF, a second with AEF and UIP and third with UIP alone. HRCT features of AEF and honeycombing were compared. There were 11 patients (10 male; mean age 69) with AEF alone, 24 patients (22 male; mean age 69) with AEF and UIP, and 15 patients (15 male; mean age 68) with UIP alone. The HRCT on the AEF alone showed subpleural (but not abutting the pleura) multiple thin-walled cysts (MTWCs) in 7 and reticular opacities in 3. The HRCT in AEF and UIP showed MTWCs in 10, reticular opacities in 17; and honeycombing in 5. Among these 35 patients with the pathological finding of AEF (with or without UIP), 17 showed MTWCs. The maximum cyst wall thickness of MTWCs (mean 0.81 mm) was significantly thinner than that of honeycombing (mean 1.56 mm). MTWCs did not locate in lung base and was distant from the pleura. HRCT findings correlated with gross findings on both cysts and honeycombing. No MTWCs were seen in the 15 patients with UIP, 8 of 15 had honeycombing on CT. We confirmed that HRCT features of AEF were MTWCs and/or reticular opacities. MTWCs might be distinguished from those of honeycomb change. While we prefer the term MTWCs, these sorts of changes have probably been confused with/interpreted as honeycombing and/or empysema in the past.


[Expression of CD44 variant exon 6 in lung cancers].
Immunohistochemical staining was carried out on 79 samples of lung cancers using anti-CD44v6 antibody. Strong positivity for CD44v6 was observed in the basal cells of bronchial epithelium, squamous metaplasia, and type II pneumocytes. The expression of CD44v6 in primary lesions of non-small cell lung cancers and small cell lung cancers was 67.6% (48/71) and 0% (0/8) respectively, while the expression of CD44v6 in metastases of lymph nodes was 57.6% (19/33) and 0% (0/5), respectively. It is suggest that expression of CD44v6 is correlated with histologic type, differentiation and metastasis of lung cancer.


Cytosolic heat-stress proteins Hsp17.7 class I and Hsp17.3 class II of tomato act as molecular chaperones in vivo.
Small heat-stress proteins (sHsps) are the most abundant stress-induced proteins with up to 20 different members in higher plants. In the cytoplasm, two different classes can be distinguished. Two cDNA clones from tomato Lycopersicon peruvianum (L.) Mill., each coding for one of the cytoplasmic sHsp subfamilies, were analyzed with respect to their transcript and protein expression, genome organization and chaperone activity. Neither type was present under control conditions but both appeared upon heat stress and in mature fruits. Expression of the class II transcript was found to be induced at slightly lower temperatures than the class I transcript. Protein analysis using class-specific antibodies revealed an identical expression pattern of both corresponding proteins. Transient expression in an Arabidopsis thaliana (L.) Heynh. cell culture showed that, despite the difference in their amino acid sequence, both classes are functionally active as chaperones in vivo, as shown by their ability to prevent thermal inactivation of firefly luciferase in a cellular environment.


Factors associated with high exclusive breastfeeding rates among preterm infants under 34 weeks of gestation in Da Nang, Vietnam: A retrospective cohort study.
Preterm infants have higher mortality than full-term infants. While breastfeeding dramatically reduces preterm death, it is limited by biological and practice barriers, particularly for babies born before 34 weeks gestational age. Da Nang Hospital for Women and Children developed a quality improvement approach to improve breastfeeding of preterm infants by strengthening feeding support, non-separation, and kangaroo mother care (KMC). To determine breastfeeding outcomes following discharge and explore factors associated with improved feeding, mothers of infants under 34 weeks gestational age born October 2021 to March 2022 and discharged alive were interviewed at six months and their medical records were reviewed. Out of 104 preterm infants included, all were exclusively breastfed at discharge and one month, 86.5% at three months, and 63.5% at six months; 47.1% received immediate skin-to-skin contact, 31.7% immediate and continuous KMC, and the remaining 68.3% continuous KMC beginning at a median of three days. Exclusive breastfeeding at six months was associated with the mother antenatally seeking breastfeeding information (odds ratio (OR) = 14.5; 95% confidence interval (CI) = 1.2-173.6), avoiding bottle-feeding at home (OR = 7.7; 95% CI = 1.7-33.7) and reduced with each day delay between birth and full breastfeeding (OR = 0.8; 95% CI = 0.6-0.9). Hospital environments that limit mother-baby separations and feeding delays, including rooming-in of mothers and infants, KMC, and breastfeeding support from birth, enabled 100% of preterm infants born before 34 weeks gestational age to breastfeed exclusively with continued rates higher than previously reported. Addressing antenatal and post-natal factors limiting practice can further improve longer-term breastfeeding outcomes. The approach can be adapted to achieve high exclusive breastfeeding rates, regardless of gestational age.


High resolution mapping of trypanosomosis resistance loci Tir2 and Tir3 using F12 advanced intercross lines with major locus Tir1 fixed for the susceptible allele.
Trypanosomosis is the most economically important disease constraint to livestock productivity in Africa. A number of trypanotolerant cattle breeds are found in West Africa, and identification of the genes conferring trypanotolerance could lead to effective means of genetic selection for trypanotolerance. In this context, high resolution mapping in mouse models are a promising approach to identifying the genes associated with trypanotolerance. In previous studies, using F2 C57BL/6J x A/J and C57BL/6J x BALB/cJ mouse resource populations, trypanotolerance QTL were mapped within a large genomic intervals of 20-40 cM to chromosomes MMU17, 5 and 1, and denoted Tir1, Tir2 and Tir3 respectively. Subsequently, using F6 C57BL/6J x A/J and C57BL/6J x BALB/cJ F6 advanced intercross lines (AIL), Tir1 was fine mapped to a confidence interval (CI) of less than 1 cM, while Tir2 and Tir3, were mapped within 5-12 cM. Tir1 represents the major trypanotolerance QTL. In order to improve map resolutions of Tir2 and Tir3, an F12 C57BL/6J x A/J AIL population fixed for the susceptible alleles at Tir1 QTL was generated. An F12 C57BL/6J x A/J AIL population, fixed for the resistant alleles at Tir1 QTL was also generated to provide an additional estimate of the gene effect of Tir1. The AIL populations homozygous for the resistant and susceptible Tir1 alleles and the parental controls were challenged with T. congolense and followed for survival times over 180 days. Mice from the two survival extremes of the F12 AIL population fixed for the susceptible alleles at Tir1 were genotyped with a dense panel of microsatellite markers spanning the Tir2 and Tir3 genomic regions and QTL mapping was performed. Tir2 was fine mapped to less than 1 cM CI while Tir3 was mapped to three intervals named Tir3a, Tir3b and Tir3c with 95% confidence intervals (CI) of 6, 7.2 and 2.2 cM, respectively. The mapped QTL regions encompass genes that are vital to innate immune response and can be potential candidate genes for the underlying QTL.


Salvage resection of advanced mediastinal tumors.
The surgical treatment of locally advanced mediastinal tumors invading the great vessels and other nearby structures still represent a tricky question, principally due to the technical complexity of the resective phase, the contingent need to carry out viable vascular reconstructions and, therefore, the proper management of pathophysiologic issues. Published large-number series providing oncologic outcomes of patients who have undergone extended radical surgery for invasive mediastinal masses are just a few. Furthermore, the wide variety of different histologies included in some of these studies, as well as the heterogeneity of chemo and radiation therapies employed, did not allow for the development of clear oncologic guidelines. Usually in the past, surgical resections of large masses along with the neighbouring structures were not offered to patients because of related morbidity and mortality and limited information available on the prognostic advantage for long term. However, in the last decades, advances in surgical technique and perioperative management, as well as increased oncologic experience in this field, have allowed radical exeresis in selected patients with invasive tumors requiring resections extended to the surrounding structures and complex vascular reconstructions. Such aggressive surgical treatment has been proposed in association or not with adjuvant chemo- or radiotherapy regimens, achieving encouraging oncologic results with limited morbidity and mortality in experienced institutions. Congestive heart failure or impending cardiovascular collapse due to the compression by the large mass are the most frequent immediately life-threatening problems that some of these patients can experience. In this setting, medical palliation is usually ineffective and an aggressive salvage surgical treatment may remain the only therapeutic option.


Factors Associated With Real-Life Functioning in Persons With Schizophrenia in a 4-Year Follow-up Study of the Italian Network for Research on Psychoses.
The goal of schizophrenia treatment has shifted from symptom reduction and relapse prevention to functional recovery; however, recovery rates remain low. Prospective identification of variables associated with real-life functioning domains is essential for personalized and integrated treatment programs. To assess whether baseline illness-related variables, personal resources, and context-related factors are associated with work skills, interpersonal relationships, and everyday life skills at 4-year follow-up. This multicenter prospective cohort study was conducted across 24 Italian university psychiatric clinics or mental health departments in which 921 patients enrolled in a cross-sectional study were contacted after 4 years for reassessment. Recruitment of community-dwelling, clinically stable persons with schizophrenia was conducted from March 2016 to December 2017, and data were analyzed from January to May 2020. Psychopathology, social and nonsocial cognition, functional capacity, personal resources, and context-related factors were assessed, with real-life functioning as the main outcome. Structural equation modeling, multiple regression analyses, and latent change score modeling were used to identify variables that were associated with real-life functioning domains at follow-up and with changes from baseline in these domains. In total, 618 participants (427 male [69.1%]; mean [SD] age, 45.1 [10.5] years) were included. Five baseline variables were directly associated with real-life functioning at follow-up: neurocognition with everyday life (β, 0.274; 95% CI, 0.207-0.341; P < .001) and work (β, 0.101; 95% CI, 0.005-0.196; P = .04) skills; avolition with interpersonal relationships (β, -0.126; 95% CI, -0.190 to -0.062; P < .001); positive symptoms with work skills (β, -0.059; 95% CI, -0.112 to -0.006; P = .03); and social cognition with work skills (β, 0.185; 95% CI, 0.088-0.283; P < .001) and interpersonal functioning (β, 0.194; 95% CI, 0.121-0.268; P < .001). Multiple regression analyses indicated that these variables accounted for the variability of functioning at follow-up after controlling for baseline functioning. In the latent change score model, higher neurocognitive abilities were associated with improvement of everyday life (β, 0.370; 95% CI, 0.253-0.486; P < .001) and work (β, 0.102; 95% CI, 0.016-0.188; P = .02) skills, social cognition (β, 0.133; 95% CI, 0.015-0.250; P = .03), and functional capacity (β, 1.138; 95% CI, 0.807-1.469; P < .001); better baseline social cognition with improvement of work skills (β, 0.168; 95% CI, 0.075-0.261; P < .001) and interpersonal functioning (β, 0.140; 95% CI, 0.069-0.212; P < .001); and better baseline everyday life skills with improvement of work skills (β, 0.121; 95% CI, 0.077-0.166; P < .001). Findings of this large prospective study suggested that baseline variables associated with functional outcome at follow-up included domains not routinely assessed and targeted by intervention programs in community mental health services. The key roles of social and nonsocial cognition and of baseline everyday life skills support the adoption in routine mental health care of cognitive training programs combined with personalized psychosocial interventions aimed to promote independent living.


Cellular substrates and laminar profile of sleep K-complex.
We describe the cellular mechanisms that underlie the generation of the K-complex, a major grapho-element of sleep electroencephalogram in humans. First we demonstrate the similarity between K-complexes recorded during natural sleep and under ketamine-xylazine anaesthesia in cats. Thereafter, we show by means of multi-site cellular and field potential recordings that K-complexes are rhythmic at frequencies of less than 1 Hz (mainly 0.5-0.9 Hz) and that they are synchronously distributed over the whole cortical surface as well as transferred to the thalamus. The surface K-complex reverses its polarity at a cortical depth of about 0.3 mm. At the cortical depth, the K-complex is made of a sharp and high-amplitude negative deflection that reflects cellular depolarization, often preceded by a smaller-amplitude, positive slow-wave reflecting cellular hyperpolarization. The sharp component of the K-complex may lead to a spindle sequence and/or to fast (mainly 20-50 Hz) oscillations. K-complexes appear spontaneously or triggered by cortical or thalamic stimulation, and they arise within cortical networks. We suggest that K-complexes, either in isolation or followed by a brief sequence of spindle waves, are the expression of the spontaneously occurring, cortically generated slow oscillation.


Identification of selenosugars and other low-molecular weight selenium metabolites in high-selenium cereal crops.
Several novel selenium containing compounds were characterized in staple crops (wheat, rice and maize) grown on soils naturally rich in selenium. A dedicated method based on the coupling of liquid chromatography with multiplexed detection (ICP-MS, ESI-Orbitrap MS(/MS)) was developed for the speciation of low-molecular weight (<5 kDa) selenium metabolites. Nine species present in different proportions as a function of the crop type were identified by cation-exchange HPLC-ESI-Orbitrap MS on the basis of the accurate molecular mass and MS/MS spectra. The natural origin of these species was then validated by varying extraction conditions and by using hydrophilic interaction LC (HILIC)-ESI-Orbitrap MS(/MS). Among the identified compounds, Se-containing monosaccharides (hexose moiety, m/z 317 and m/z 358) or Se-containing disaccharides (hexose-pentose moiety, m/z 407 and m/z 408) were the first selenosugars reported in edible plants. It is also the first report of the presence of 2,3-dihydroxypropionyl-selenolanthionine (m/z 345) in rice. Because these crops can be an important source of selenium in animal and human nutrition, the understanding of the origin and the fate of these species during metabolic processes will be of great interest.


Cathepsin D serum levels are not a valid serum marker in renal cell carcinoma.
It was the aim of this study to evaluate the serum levels of cathepsin D (Cath D) as a diagnostic tool in patients with renal cell carcinoma (RCC) in comparison with healthy volunteers. Cath D serum levels were measured in serum samples obtained preoperatively from 32 patients with histologically confirmed RCC versus 30 healthy individuals using an enzyme immunoassay. Additionally, for the tumor group, Cath D serum levels were correlated with tumor stage and grade as determined according to the 2002 TNM classification. The serum Cath D concentration was not significantly different in patients with RCC compared with healthy individuals (mean 16.58 vs. 16.64 ng/ml; p = 0.43). Furthermore, there was no significant association between Cath D serum levels and several patient or tumor characteristics such as tumor stage, tumor grade, lymph node status, presence of metastasis, gender or age. In contrast to overexpression of Cath D in primary RCC tissue, serum Cath D is not altered in RCC patients when compared with healthy volunteers. In this small cohort, Cath D serum levels did not reveal additional clinical information in patients diagnosed with a small renal mass. Further prospective multicenter studies might shed more light on the value of Cath D in the diagnostics of RCC.


Effects of tacrolimus hydrate (FK-506) on the expression of tissue factor in THP-1 human monocyte cell line.
Accelerated coronary atherosclerosis in cardiac allografts is a major factor limiting survival after heart transplantation, and activation of the coagulation system contributes to accelerated transplant atherosclerosis. Accordingly, increased tissue factor (TF) expression by monocyte/macrophages may play a pivotal role underlying deposition of fibrin in the affected vessels. To evaluate the potential effects of an important immunosuppressive agent, tacrolimus hydrate (FK-506), on monocyte/macrophages and their response to lipopolysaccharide (LPS), we exposed human monocyte/macrophage cell line (THP-1 cells), to LPS and characterized its procoagulant activity (PCA). FK-506 exerted a concentration-dependent inhibitory effect on LPS (10 micrograms/ml) induction of procoagulant activity, identified as TF activity as judged from immunostaining of TF antigen and by functional characterization with the use of coagulation factor VII-deficient plasma and an antibody against human TF. In addition, the reverse transcription polymerase chain reaction demonstrated reduced expression of TF mRNA in LPS-stimulated THP-1 cells exposed to FK-506. Thus, FK-506 acts favorably not only as a direct immunomodulating agent but also as an alleviator of local activation of the coagulation cascade contributing to transplant arteriopathy through modulation of monocyte expression of TF.


On the regulation of protein synthesis in vaccinia virus infected cells.
All eukaryotic mRNA species show a characteristic individual translational efficiency under conditions of restricted polypeptide chain initiation caused by an increase in the osmolarity of the growth medium. In vaccinia virus infected L cells or HeLa cells virus mRNAs can be grouped into classes on the basis of their relative labelling under standard and hypertonic conditions. Under the latter conditions, most of the "early" mRNAs possess very high translational efficiencies, most of the "intermediate" mRNAs show an intermediate efficiency and the most prominent "late" mRNAs show a translational efficiency which is lower than that of other virus mRNAs but still higher than the average cellular mRNA. Late in the infection cycle virus mRNAs with a relative low translational efficiency are preferentially translated under standard growth conditions whereas "early" virus mRNAs which are still present and which show a higher translational resistance to hypertonic conditions are not translated. These results indicate a unique translational control operating late in the growth cycle of vaccinia virus.


Feasibility of in situ hybridisation with chromosome specific DNA probes on paraffin wax embedded tissue.
The feasibility was studied of in situ hybridisation using chromosome specific DNA probes on paraffin wax embedded normal and malignant tissues from different organs. Both isolated nuclei and 5 microns sections were used in in situ hybridisation experiments with biotinylated repetitive DNA probes specific for the centromeric regions of chromosomes 1 and 17. The hybridisation results were visualised with peroxidase-diaminobenzidine. The optimal pretreatments with sodium thiocyanate and pepsin were experimentally defined for the different tissues. Although interphase cytogenetics on paraffin wax embedded tissue is possible, the results indicate that it has its limitations, compared with investigations on fresh tumour tissue.


The genome of the white-rot fungus Pycnoporus cinnabarinus: a basidiomycete model with a versatile arsenal for lignocellulosic biomass breakdown.
Saprophytic filamentous fungi are ubiquitous micro-organisms that play an essential role in photosynthetic carbon recycling. The wood-decayer Pycnoporus cinnabarinus is a model fungus for the study of plant cell wall decomposition and is used for a number of applications in green and white biotechnology. The 33.6 megabase genome of P. cinnabarinus was sequenced and assembled, and the 10,442 predicted genes were functionally annotated using a phylogenomic procedure. In-depth analyses were carried out for the numerous enzyme families involved in lignocellulosic biomass breakdown, for protein secretion and glycosylation pathways, and for mating type. The P. cinnabarinus genome sequence revealed a consistent repertoire of genes shared with wood-decaying basidiomycetes. P. cinnabarinus is thus fully equipped with the classical families involved in cellulose and hemicellulose degradation, whereas its pectinolytic repertoire appears relatively limited. In addition, P. cinnabarinus possesses a complete versatile enzymatic arsenal for lignin breakdown. We identified several genes encoding members of the three ligninolytic peroxidase types, namely lignin peroxidase, manganese peroxidase and versatile peroxidase. Comparative genome analyses were performed in fungi displaying different nutritional strategies (white-rot and brown-rot modes of decay). P. cinnabarinus presents a typical distribution of all the specific families found in the white-rot life style. Growth profiling of P. cinnabarinus was performed on 35 carbon sources including simple and complex substrates to study substrate utilization and preferences. P. cinnabarinus grew faster on crude plant substrates than on pure, mono- or polysaccharide substrates. Finally, proteomic analyses were conducted from liquid and solid-state fermentation to analyze the composition of the secretomes corresponding to growth on different substrates. The distribution of lignocellulolytic enzymes in the secretomes was strongly dependent on growth conditions, especially for lytic polysaccharide mono-oxygenases. With its available genome sequence, P. cinnabarinus is now an outstanding model system for the study of the enzyme machinery involved in the degradation or transformation of lignocellulosic biomass.


Chryseobacterium binzhouense sp. nov., isolated from activated sludge.
A golden-pigmented, Gram-strain-negative, aerobic, rod-shaped, non-flagellated and non-gliding bacterium, designated strain lm2T, was isolated from activated sludge obtained from a wastewater treatment plant in Binzhou (Shandong province, PR China). Growth occurred at 15-45°C (optimum, 30 °C), in the presence of 0-5.0 % (w/v) NaCl (optimum, 0-2.0 %) and at pH 6.5-8.0 (optimum, pH 7.0-7.5). The chemotaxonomic, phenotypic and genomic traits were investigated. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain lm2T belonged to the genus Chryseobacterium, with highest sequence similarity to Chryseobacterium echinoideorum CC-CZW010T (97.1 %). Genome sequencing revealed a genome size of 3 611 894 bp and a G+C content of 34.9 mol%. The average nucleotide identity value and the digital DNA-DNA hybridization (dDDH) value between strain lm2T and C. echinoideorum JCM 30470T were 87.8 and 34.7 %, respectively. The major respiratory quinone was Menaquinone-6 (MK-6). The major fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH and iso-C17 : 1 ω9c and its polar lipids consisted of phosphatidylethanolamine (PE), unidentified lipids (L1-5) and unidentified aminolipids (AL1-4). On the basis of these data, strain lm2T is considered to represent a novel species of the genus Chryseobacterium, for which the name Chryseobacterium binzhouense sp. nov. is proposed. The type strain is lm2T (=KCTC 72529T=CCTCC AB2019126T).


Exome analysis focusing on epilepsy-related genes in children and adults with sudden unexplained death.
Genetic studies in sudden infant death syndrome (SIDS) and sudden unexplained death (SUD) cohorts have indicated that cardiovascular diseases might have contributed to sudden unexpected death in 20-35 % of autopsy-negative cases. Sudden unexpected death can also occur in people with epilepsy, termed as sudden unexpected death in epilepsy (SUDEP). The pathophysiological mechanisms of SUDEP are not well understood, but are likely multifactorial, including seizure-induced hypoventilation and arrhythmias as well as genetic risk factors. The sudden death of some of the SIDS/SUD victims might also be explained by genetic epilepsy, therefore this study aimed to expand the post-mortem genetic analysis of SIDS/SUD cases to epilepsy-related genes. Existing whole-exome sequencing data from our 155 SIDS and 45 SUD cases were analyzed, with a focus on 365 epilepsy-related genes. Nine of the SUD victims had a known medical history of epilepsy, seizures or other underlying neurological conditions and were therefore classified as SUDEP cases. In our SIDS and SUD cohorts, we found epilepsy-related pathogenic/likely pathogenic variants in the genes OPA1, RAI1, SCN3A, SCN5A and TSC2. Post-mortem analysis of epilepsy-related genes identified potentially disease-causing variants that might have contributed to the sudden death events in our SIDS/SUD cases. However, the interpretation of identified variants remains challenging and often changes over time as more data is gathered. Overall, this study contributes insight in potentially pathophysiological epilepsy-related mechanisms in SIDS, SUD and SUDEP victims and underlines the importance of sensible counselling on the risk and preventive measures in genetic epilepsy.


Experience with totally laparoscopic distal pancreatectomy with splenic preservation for pediatric trauma--2 techniques.
Blunt pancreatic traumatic injury in children, although rare, can be managed with a variety of methods from nonoperative, early operative, or delayed operative strategies. In the appropriate setting, early operative intervention has been associated with shorter hospitalization and decreased morbidity for these patients. Case reports describe laparoscopic distal pancreatectomy for isolated pancreatic laceration in children. This article presents the experience and results of the first series of totally laparoscopic, spleen-preserving distal pancreatectomies for trauma in children. Three children aged 8 to 13 years underwent laparoscopic distal pancreatectomy with splenic preservation for traumatic pancreatic transection within 72 hours of initial injury. Computed tomography imaging in all patients demonstrated complete pancreatic transection. The details of 2 operative techniques used for totally laparoscopic distal pancreatectomy are described. The data for associated injuries, amylase/lipase levels, operative management, postoperative course, length of stay, complications, and follow-up were collected for all patients. All 3 children aged 8, 10, and 13 years underwent laparoscopic distal pancreatectomy without splenectomy within 72 hours of injury (23, 48, and 72 hours). The mechanism of injury was from a bicycle handle, knee to abdomen, and dirt bike handle, respectively. The length of hospital stay was 6, 15, and 7 days with follow-up of 12, 35, and 34 months. The 2 older children underwent pancreatic transection with an endostapler, and the 8-year-old had the pancreatic remnant oversewn by hand. Use of postoperative total parenteral nutrition continued for 0, 13, and 7 days. Complications included an abdominal wall hematoma and prolonged ileus with mild pancreatitis. There were no pancreatic fistulae or insufficiency. All patients are doing well and are asymptomatic from prior injury and laparoscopic distal pancreatectomy. In the appropriate pediatric patient with traumatic pancreatic transection, a laparoscopic distal pancreatectomy with splenic preservation can be performed safely, with low morbidity and good outcomes. Further studies with larger series of patients with these injuries would be useful.