Adherence to Sulfur Microbial Diet and Ovarian Cancer Survival: Evidence from a Prospective Cohort Study.
The study aims to investigate the role of the sulfur microbial diet in the survival of ovarian cancer (OC). A prospective cohort study is conducted with 703 patients diagnosed with OC between 2015 and 2020. Diet information is collected using a validated food frequency questionnaire. Deaths are ascertained up to March 31, 2021, via the death registry linkage. During the follow-up period (median: 37.2 months, interquartile range: 24.7-50.2 months), 130 deaths are observed. A higher sulfur microbial diet score is significantly associated with an increased risk of all-cause mortality among OC patients (tertile 3 vs tertile 1: HR = 1.93, 95% CI = 1.11-3.35). Each 1-standard deviation increment in the sulfur microbial diet score increases the all-cause mortality risk by 33% (95% CI = 1.04-1.71). Stratified analysis shows that significant associations are found in OC patients diagnosed over 50 years of age, with body mass index ≥24 kg m-2 , who changed their diet after diagnosis, or without residual lesions. Adherence to the sulfur microbial diet, characterized by high intakes of red meats and processed meats, and low intakes of fruits, vegetables, and whole grains, is associated with poor survival in OC patients.


Pediatric tracheotomy: indications and decannulation outcomes.
The objective of this study was to determine if there are differences in decannulation rates and duration of cannulation between pediatric patients undergoing tracheotomy for different indications. Retrospective chart review. Medical records for pediatric patients (age 0-18 years) undergoing tracheotomy between January 1, 2003, and May 31, 2012, were retrospectively reviewed. Patients were assigned an indication for tracheotomy from five categories: neurological, cardiopulmonary, upper airway obstruction, craniofacial anomalies, and maxillofacial/laryngotracheal trauma. Initial chart review identified 124 patients, 113 for whom complete data was available. Of these patients, the indications for tracheotomy were cardiopulmonary disease in 24 (21.2%), craniofacial anomalies in 12 (10.6%), neurological impairment in 44 (38.9%), traumatic injury in 11 (9.7%), and upper airway obstruction in 22 (19.5%). The time to decannulation was shorter for trauma patients compared to cardiopulmonary (P = 0.044) and neurological patients (P = 0.001). A total of 32 (31.9%) patients were decannulated during the study period, with a higher rate in trauma patients (72.7%) and a lower rate in those with upper airway obstruction (36.4%) than would be expected under homogeneity. Of the 32 patients who were decannulated, 11 (30.6%) were decannulated during the same hospitalization in which the tracheotomy was performed. This study demonstrates a difference in overall decannulation rates and a shorter time to decannulation in children undergoing tracheotomy for maxillofacial and laryngotracheal trauma compared to cardiopulmonary and neurological indications.


The epidemiology of acute respiratory distress syndrome in patients presenting to the emergency department with severe sepsis.
Acute respiratory distress syndrome (ARDS) is a serious complication of sepsis, and sepsis-associated ARDS is associated with significant morbidity and mortality. To date, no study has directly examined the epidemiology of ARDS in severe sepsis from the earliest presentation to the health care system, the emergency department (ED). This was a single-center retrospective, observational cohort study of 778 adults with severe sepsis presenting to the ED. The primary outcome was the development of ARDS requiring mechanical ventilation during the first 5 hospital days. Acute respiratory distress syndrome was defined using the Berlin definition. We used multivariable logistic regression to identify risk factors associated independently with ARDS development. The incidence of ARDS was 6.2% (48/778 patients) in the entire cohort. Acute respiratory distress syndrome development varied across the continuum of care: 0.9% of patients fulfilled criteria for ARDS in the ED, 1.4% admitted to the ward developed ARDS, and 8.9% admitted to the intensive care unit developed ARDS. Acute respiratory distress syndrome developed a median of 1 day after admission and was associated with a 4-fold higher risk of in-hospital mortality (14% vs. 60%, P < 0.001). Independent risk factors associated with increased risk of ARDS development included intermediate (2-3.9 mmol/L) (P = 0.04) and high (≥4) serum lactate levels (P = 0.008), Lung Injury Prediction score (P < 0.001), and microbiologically proven infection (P = 0.01). In patients presenting to the ED with severe sepsis, the rate of sepsis-associated ARDS development varied across the continuum of care. Acute respiratory distress syndrome developed rapidly and was associated with significant mortality. Elevated serum lactate levels in the ED and a recently validated clinical prediction score were independently associated with the development of ARDS in severe sepsis.


Equal access to healthcare does not eliminate disparities in the management of adults with appendicitis.
Race and socioeconomic status have led to inequality in access to surgical care, leading to differences in appendiceal perforation rates. However, in a setting of equal health care access, these disparities were not evident. Currently, disparities exist with use of laparoscopic appendectomy (LA) in the treatment of appendicitis. This study determines whether equal health care access eliminates differences in the management and outcomes of appendicitis. A retrospective review from 1998 to 2007 was performed. All members in this single-provider system have equal healthcare access. Socioeconomic data were extracted from the US Census database. Study outcome was use of LA and postoperative morbidity. Independent variables included age (18-44 y, 45-65 y, and >65 y), gender, race, annual mean household income (<40 K, 40-65 K, and >65 K), and education level. Univariate and multivariable (controlling for age, gender, perforation, race/ethnicity, income, and education level) analyses were performed. A total of 16,196 patients were identified (mean age = 41 y, 54% male). Sixty percent of patients underwent LA. On multivariable analysis, male gender was associated with a decreased use of LA (OR 0.84, CI 0.79-0.89, P < 0.0001). There was a lower use of LA in older patients (age > 65 y) and patients age 45-65 y compared with younger patients (18-44 y). There was a lower use of LA in Blacks and a higher rate in Hispanics compared with Whites. Use of LA was higher in high income and middle income patients compared with low income patients. The use of LA was similar across all education levels. Postoperative morbidity was higher in males, older patients (age > 65 y), and patients age 45-65 y. However, postoperative morbidity was similar across all race/ethnic groups and all income levels and education levels. In a setting of equal health care access, there is evidence of gender, racial, age, and socioeconomic disparities for use of LA and worse outcomes for males and older patients. These data suggest that despite equal healthcare access, disparities continue to exist with respect to race and income level for the management of adults with appendicitis.


Navigating the therapeutic complexity of PI3K pathway inhibition in melanoma.
Melanoma is entering into an era of combinatorial approaches to build upon recent clinical breakthroughs achieved by novel single-agent therapies. One of the leading targets to emerge from the growing understanding of the molecular pathogenesis, heterogeneity, and resistance mechanisms of melanomas is the phosphoinositide 3-kinase (PI3K)-AKT pathway. Multiple genetic and epigenetic aberrations that activate this pathway have been identified in melanomas de novo and in acquired resistance models. These developments have been paralleled by the establishment of models for preclinical testing and the availability of compounds that target various effectors in the pathway. Thus, in addition to having a strong rationale for targeting, the PI3K-AKT pathway presents an immediate clinical opportunity. However, the development of effective strategies against this pathway must overcome several key challenges, including optimizing patient selection, overcoming feedback loops, and pathway cross-talk that can mediate resistance. This review discusses the current understanding and ongoing research about the PI3K-AKT pathway in melanoma and emerging strategies to achieve clinical benefit in patients by targeting it.


Clonal character of F1 hybrid lymphocyte subset recognition of parental cells in one-way mixed lymphocyte cultures.
Proliferation of F(1) hybrid lymphocytes in mixed lymphocyte cultures is stimulated by mitomycin-blocked parental cells. The demonstration of this phenomenon using F(1) hybrids derived from congenic lines of mice establishes that the stimulation is controlled by genes in or closely linked to the major histocompatibility locus chromosome region. In agreement with the finding that tumor-bearing mice have an increased capacity for primary alloantigen recognition, it was observed that the F(1) hybrid response to parent was also augmented by tumor bearing. Chromosomal analysis of dividing cells in one-way mixed cultures confirms that F(1) cells, and not the blocked parental cells, enter mitosis. Stimulation of F(1) cells by a soluble mediator liberated by the parental cells was not observed and mitomycin blocking of parental cells seems to be a completely effective blocking agent ensuring that parental cells can not enter DNA synthesis. The specificity and clonal nature of F(1) recognition of parent was demonstrated using a 5-bromodeoxyuridine-suicide procedure. Distinct clones of lymphocytes in F(1) spleen cell populations seem to recognize one or the other parent, but not both, in such experiments. These observations and others in tumor systems suggest that most or all heterozygous organisms may possess potentially self-reactive clones of lymphocytes.


Implantable cardioverter-defibrillator in hypertrophic cardiomyopathy.
Sudden cardiac death (SCD) is the most devastating complication in hypertrophic cardiomyopathy (HCM). The implantable cardioverter-defibrillator (ICD) has proven to be effective in SCD prevention in several clinical scenarios. In HCM population, it has demonstrated to successfully abort life-threatening ventricular arrhythmias despite the extreme morphology characteristic of HCM, often with massive degrees of left ventricular hypertrophy and/or LV outflow tract obstruction. Studies showed a high rate of appropriate intervention in secondary prevention and in primary prevention of patients considered at high risk. This appropriate intervention rate is even more significant considering the young and otherwise healthy patients that compose HCM population. Since SCD incidence in HCM is relatively low, optimal identification of patients at high risk is crucial. Classical strategy of risk stratification based on clinical risk factors has several limitations and has proven to overestimate risk. A new risk prediction model that provides individual 5-year estimated risk appears to be superior to traditional models based on bivariate risk factors. Perioperative complications seem to be similar to those related to the implant of other cardiac devices, while long-term complications have been traditionally in the spotlight. HCM patients are considered more vulnerable to ICD-related complications and inappropriate ICD therapy because of their young age at implant and increased prevalence of atrial fibrillation, but long-term follow-up data on ICD-related complications in general practice is limited. The subcutaneous implantable cardioverter defibrillator seems to be a safe and effective alternative in HCM, although long-term data are scarce.


Spondyloenchondrodysplasia: clinical variability in three cases.
Spondyloenchondrodysplasia is a very rare skeletal dysplasia in which multiple enchondromata exist in the metaphyses of the long bones with platyspondyly. We present three patients (two of them are sibs) with spondyloenchondrodysplasia. The first patient was a 10-year-old boy, who had short stature and enchondromatous-like lesions in the metaphyses of the long bones and platyspondyly on radiography. His older sister (21-years old) had received growth hormone therapy 12 years earlier due to short stature, and her radiological findings were similar but milder than her brother. Both the sibs had normal intelligence and no calcification of the basal ganglia. The third patient was a 6-year-old boy who had short stature, mental retardation, enchondromatous-like lesions in the metaphyses of the long bones and platyspondyly. His cranial BT showed calcification of basal ganglia. The findings of the two sibs in the first family were compatible with spondyloenchondrodysplasia. The difference in clinical severity between the siblings shows the variability within the family. The third case with mental retardation and the presence of intracranial calcifications is compatible with spondyloenchondrodysplasia with basal ganglia calcification. In conclusion, we suggest that family screening and cranial imaging for the presence of intracranial calcifications should be considered in every patient with the diagnosis of spondyloenchondrodysplasia.


Muscle fatigue in spinal muscular atrophy.
We previously reported that patients with spinal muscular atrophy do not lose muscle strength over time as measured quantitatively. However, we noted that many patients with spinal muscular atrophy suffer from what they call fatigue. We wondered if we could measure fatigue during a single maximal voluntary contraction, whether fatigue might increase with time, independent of muscle strength, and whether increasing fatigue might correlate with loss of function in some patients. We measured fatigue during a single maximal voluntary contraction in a cohort of patients having spinal muscular atrophy using quantitative strength testing. We included only patients with spinal muscular atrophy aged 5 years or older, so they could follow instructions regarding muscle contraction, and who were followed for at least 2 years. Seventy-six children with spinal muscular atrophy and 24 untrained individuals, aged 5 to 57 years (mean, 16.8 years), were studied. There was no discernible abnormal fatigue in patients with spinal muscular atrophy compared to untrained controls using our methodology. Thus, spinal muscular atrophy may not be associated with fatiguability. Moreover, spinal muscular atrophy does not appear to cause progressive muscle fatigue with age or loss of function. It is possible that fatigue was undetectable by our methods. An alternative explanation is that what patients describe as fatigue may be caused by factors outside the neuromuscular system. Such factors may include chronic respiratory insufficiency with hypoventilation and carbon dioxide retention as well as chronic malnutrition and negative nitrogen balance.


All-Cellulose Composite Laminates Made from Wood-Based Textiles: Effects of Process Conditions and the Addition of TEMPO-Oxidized Nanocellulose.
All-cellulose composites (ACCs) are manufactured using only cellulose as a raw material. Biobased materials are more sustainable alternatives to the petroleum-based composites that are used in many technical and life-science applications. In this study, an aquatic NaOH-urea solvent system was used to produce sustainable ACCs from wood-based woven textiles with and without the addition of TEMPO-oxidized nanocellulose (at 1 wt.-%). This study investigated the effects of dissolution time, temperature during hot press, and the addition of TEMPO-oxidized nanocellulose on the mechanical and thermal properties of the composites. The results showed a significant change in the tensile properties of the layered textile composite at dissolution times of 30 s and 1 min, while ACC elongation was the highest after 2 and 5 min. Changes in hot press temperature from 70 °C to 150 °C had a significant effect: with an increase in hot press temperature, the tensile strength increased and the elongation at break decreased. Incorporating TEMPO-oxidized nanocellulose into the interface of textile layers before partial dissolution improved tensile strength and, even more markedly, the elongation at break. According to thermal analyses, textile-based ACCs have a higher storage modulus (0.6 GPa) and thermal stabilization than ACCs with nanocellulose additives. This study highlights the important roles of process conditions and raw material characteristics on the structure and properties of ACCs.


Antiretroviral drug resistance testing in adult HIV-1 infection: recommendations of an International AIDS Society-USA Panel.
Assays for drug resistance testing in human immunodeficiency virus type 1 (HIV-1) infection are now available and clinical studies suggest that viral drug resistance is correlated with poor virologic response to new therapy. The International AIDS Society-USA sought to update prior recommendations to provide guidance for clinicians regarding indications for HIV-1 resistance testing. An International AIDS Society-USA 13-member physician panel with expertise in basic science, clinical research, and patient care involving HIV resistance to antiretroviral drugs was reconvened to provide recommendations for the clinical use of drug resistance testing. The full panel met regularly between January and October 1999. Resistance and resistance testing data appearing in the last decade through April 2000 and presentations at national and international research conferences were reviewed. Recommendations and considerations were developed by 100% group consensus, acknowledging that definitive data to support final recommendations are not yet available. Emerging data indicate that despite limitations, resistance testing should be incorporated into patient management in some settings. Resistance testing is recommended to help guide the choice of new regimens after treatment failure and for guiding therapy for pregnant women. It should be considered in treatment-naive patients with established infection, but cannot be firmly recommended in this setting. Testing also should be considered prior to initiating therapy in patients with acute HIV infection, although therapy should not be delayed pending the results. Expert interpretation is recommended given the complexity of results and assay limitations.


High-efficiency cloning of full-length cDNA.
A widely recognized difficulty of presently used methods for cDNA cloning is obtaining cDNA segments that contain the entire nucleotide sequence of the corresponding mRNA. The cloning procedure described here mitigates this shortcoming. Of the 10(5) plasmid-cDNA recombinants obtained per microgram of rabbit reticulocyte mRNA, about 10% contained a complete alpha- of beta-globin mRNA sequence, and at least 30 to 50%, but very likely more, contained the entire globin coding regions. We attribute the high efficiency of cloning full- or nearly full-length cDNA to (i) the fact that the plasmid DNA vector itself serves as the primer for first- and second-strand cDNA synthesis, (ii) the lack of any nuclease treatment of the products, and (iii) the fact that one of the steps in the procedure results in preferential cloning of recombinants with full-length cDNA's over those with truncated cDNA's.


Biocompatible Near-Infrared Three-Dimensional Tracking System.
A fundamental challenge in soft-tissue surgery is that target tissue moves and deforms, becomes occluded by blood or other tissue, and is difficult to differentiate from surrounding tissue. We developed small biocompatible near-infrared fluorescent (NIRF) markers with a novel fused plenoptic and NIR camera tracking system, enabling three-dimensional tracking of tools and target tissue while overcoming blood and tissue occlusion in the uncontrolled, rapidly changing surgical environment. In this work, we present the tracking system and marker design and compare tracking accuracies to standard optical tracking methods using robotic experiments. At speeds of 1 mm/s, we observe tracking accuracies of 1.61 mm, degrading only to 1.71 mm when the markers are covered in blood and tissue.


A representative value for whole-day BP monitoring.
Shorter-term alternatives to 24-hour monitoring of BP were analyzed in six hypertensive patients. Average BPs during the 24-hour period were lower than averages during a two-hour morning period or those based on either three readings or a single reading taken at 8 AM. The systolic and diastolic BP two-hour averages correlated more strongly with the 24-hour averages than did the three-reading or single-reading alternatives. Thus, the average of serial BP readings obtained during a two-hour morning period is a consistent predictor of the whole-day BP and represents it more closely than the conventionally used single reading.


Npa1p is an essential trans-acting factor required for an early step in the assembly of 60S ribosomal subunits in Saccharomyces cerevisiae.
Ribosome biogenesis requires >100 nonribosomal proteins, which are associated with different preribosomal particles. The substrates, the interacting partners, and the timing of action of most of these proteins are largely unknown. To elucidate the functional environment of the putative ATP-dependent RNA helicase Dbp6p from Saccharomyces cerevisiae, which is required for 60S ribosomal subunit assembly, we have previously performed a synthetic lethal screen and thereby revealed a genetic interaction network between Dbp6p, Rpl3p, Nop8p, and the novel Rsa3p. In this report, we extended the characterization of this functional network by performing a synthetic lethal screen with the rsa3 null allele. This screen identified the so far uncharacterized Npa1p (YKL014C). Polysome profile analysis indicates that there is a deficit of 60S ribosomal subunits and an accumulation of halfmer polysomes in the slowly growing npa1-1 mutant. Northern blotting and primer extension analysis shows that the npa1-1 mutation negatively affects processing of all 27S pre-rRNAs and the normal accumulation of both mature 25S and 5.8S rRNAs. In addition, 27SA(2) pre-rRNA is prematurely cleaved at site C(2). Moreover, GFP-tagged Npa1p localizes predominantly to the nucleolus and sediments with large complexes in sucrose gradients, which most likely correspond to pre-60S ribosomal particles. We conclude that Npa1p is required for ribosome biogenesis and operates in the same functional environment of Rsa3p and Dbp6p during early maturation of 60S ribosomal subunits.


Targeted disruption of the class B scavenger receptor CD36 protects against atherosclerotic lesion development in mice.
Macrophage scavenger receptors have been implicated as key players in the pathogenesis of atherosclerosis. To assess the role of the class B scavenger receptor CD36 in atherogenesis, we crossed a CD36-null strain with the atherogenic apo E-null strain and quantified lesion development. There was a 76.5% decrease in aortic tree lesion area (Western diet) and a 45% decrease in aortic sinus lesion area (normal chow) in the CD36-apo E double-null mice when compared with controls, despite alterations in lipoprotein profiles that often correlate with increased atherogenicity. Macrophages derived from CD36-apo E double-null mice bound and internalized more than 60% less copper-oxidized LDL and LDL modified by monocyte-generated reactive nitrogen species. A similar inhibition of in vitro lipid accumulation and foam cell formation after exposure to these ligands was seen. These results support a major role for CD36 in atherosclerotic lesion development in vivo and suggest that blockade of CD36 can be protective even in more extreme proatherogenic circumstances.


A quantitative proteomic approach for identification of potential biomarkers in hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide. In this study, our objective was to identify differentially regulated proteins in HCC through a quantitative proteomic approach using iTRAQ. More than 600 proteins were quantitated of which 59 proteins were overexpressed and 92 proteins were underexpressed in HCC as compared to adjacent normal tissue. Several differentially expressed proteins were not implicated previously in HCC. A subset of these proteins (six each from upregulated and downregulated groups) was further validated using immunoblotting and immunohistochemical labeling. Some of the overexpressed proteins with no previous description in the context of HCC include fibroleukin, interferon induced 56 kDa protein, milk fat globule-EGF factor 8, and myeloid-associated differentiation marker. Interestingly, all the enzymes of urea metabolic pathway were dramatically downregulated. Immunohistochemical labeling confirmed differential expression of fibroleukin, myeloid associated differentiation marker and ornithine carbamoyl transferase in majority of HCC samples analyzed. Our results demonstrate quantitative proteomics as a robust discovery tool for the identification of differentially regulated proteins in cancers.


A conserved determinant in the V1 loop of HIV-1 modulates the V3 loop to prime low CD4 use and macrophage infection.
The CD4 binding site (CD4bs) on the HIV-1 envelope plays a major role in determining the capacity of R5 viruses to infect primary macrophages. Thus, envelope determinants within or proximal to the CD4bs have been shown to control the use of low CD4 levels on macrophages for infection. These residues affect the affinity for CD4 either directly or indirectly by altering the exposure of CD4 contact residues. Here, we describe a single amino acid determinant in the V1 loop that also modulates macrophage tropism. Thus, we identified an E153G substitution that conferred high levels of macrophage infectivity for several heterologous R5 envelopes, while the reciprocal G153E substitution abrogated infection. Shifts in macrophage tropism were associated with dramatic shifts in sensitivity to the V3 loop monoclonal antibody (MAb), 447-52D and soluble CD4, as well as more modest changes in sensitivity to the CD4bs MAb, b12. These observations are consistent with an altered conformation or exposure of the V3 loop that enables the envelope to use low CD4 levels for infection. The modest shifts in b12 sensitivity suggest that residue 153 impacts on the exposure of the CD4bs. However, the more intense shifts in sCD4 sensitivity suggest additional mechanisms that likely include an increased ability of the envelope to undergo conformational changes following binding to suboptimal levels of cell surface CD4. In summary, we show that a conserved determinant in the V1 loop modulates the V3 loop to prime low CD4 use and macrophage infection.


Endometriosis in abdominal scars: a diagnostic pitfall.
Endometriosis is ectopic endometrial tissue that responds to hormonal stimulation and is found in 8 to 15 per cent of all menstruating women. Endometrioma in a surgical scar is rare and appears in 0.1 per cent of women who have undergone cesarean section; 25 per cent of these women have concomitant pelvic endometriosis. Endometrioma in a scar can present as a painful swelling on the scar that worsens during cough and effort, with the patient's complaints resembling those of a postoperative hernia occurring in a scar. It can also mimic other lesions of the abdominal wall, such as hematomas, granulomas, abscesses, and tumors. Four cases of endometriomas in a cesarean section scar are described. Two were diagnosed preoperatively as postoperative ventral hernias, and the other two presented with an abdominal wall mass causing pain and discomfort. All patients underwent surgery, and the error of their preoperative diagnosis was revealed by histology, which confirmed endometrioma. We contend that endometrioma in scarring is a diagnostic pitfall that should be considered in the differential diagnosis of postoperative ventral hernias and various abdominal wall masses.


All-Solution-Processed Thermally and Chemically Stable Copper-Nickel Core-Shell Nanowire-Based Composite Window Electrodes for Perovskite Solar Cells.
Organic-inorganic hybrid perovskite solar cells (PSCs) have recently attracted tremendous attention because of their excellent efficiency and the advantage of a low-cost fabrication process. As a transparent electrode for PSCs, the application of copper nanowire (CuNW)-network was limited because of its thermal/chemical instability, despite its advantages in terms of high optical/electrical properties and low-cost production. Here, the copper-nickel core-shell nanowire (Cu@Ni NW)-based composite electrode is proposed as a bottom window electrode for PSCs, without the involvement of a high-cost precious metal and vacuum process. The dense and uniform Ni protective shell for CuNWs is attainable by simple electroless plating, and the resulting Cu@Ni NWs exhibit outstanding chemical stability as well as thermal stability compared with bare CuNWs. When the Ni layer with the optimal thickness is introduced, the Cu@Ni NW electrode shows a high transmittance of 80.5% AVT at 400-800 nm, and a sheet resistance of 49.3 ± 5 Ω sq-1. Using the highly stable Cu@Ni NWs, the composite electrode structure is fabricated with sol-gel-derived Al-doped zinc oxide (AZO) over-layer for better charge collection and additional protection against iodine ions from the perovskite. The PSCs fabricated with AZO/Cu@Ni NW-based composite electrode demonstrate a power conversion efficiency (PCE) of 12.2% and excellent long-term stability maintaining 91% of initial PCE after being stored for 500 h at room temperature. Experimental results demonstrate the potential of highly stable Cu@Ni NW-based electrodes as the cost-effective alternative transparent electrode, which can facilitate the commercialization of PSCs.


Household pesticides and risk of pediatric brain tumors.
A follow-up to a population-based case-control study of pediatric brain tumors in Los Angeles County, California, involving mothers of 224 cases and 218 controls, investigated the risk of household pesticide use from pregnancy to diagnosis. Risk was significantly elevated for prenatal exposure to flea/tick pesticides -odds ratio (OR) = 1.7; 95% confidence interval (CI), 1.1-2.6-, particularly among subjects less than 5 years old at diagnosis (OR = 2.5; CI, 1. 2-5.5). Prenatal risk was highest for mothers who prepared, applied, or cleaned up flea/tick products themselves (OR = 2.2; CI, 1.1-4.2; for subjects <5 years of age, OR = 5.4; CI, 1.3-22.3). A significant trend of increased risk with increased exposure was observed for number of pets treated (p = 0.04). Multivariate analysis of types of flea/tick products indicated that sprays/foggers were the only products significantly related to risk (OR =10.8; CI, 1.3-89.1). Elevated risks were not observed for termite or lice treatments, pesticides for nuisance pests, or yard and garden insecticides, herbicides, fungicides, or snail killer. Certain precautions,if ignored, were associated with significant increased risk: evacuating the house after spraying or dusting for pests (OR = 1.6; CI, 1.0-2.6), delaying the harvest of food after pesticide treatment (OR = 3.6; CI, 1.0-13.7), and following instructions on pesticide labels (OR = 3. 7;CI, 1.5-9.6). These findings indicate that chemicals used in flea/tick products may increase risk of pediatric brain tumors and suggest that further research be done to pinpoint specific chemicals involved.


Statistical methods for temporal and space-time analysis of community composition data.
This review focuses on the analysis of temporal beta diversity, which is the variation in community composition along time in a study area. Temporal beta diversity is measured by the variance of the multivariate community composition time series and that variance can be partitioned using appropriate statistical methods. Some of these methods are classical, such as simple or canonical ordination, whereas others are recent, including the methods of temporal eigenfunction analysis developed for multiscale exploration (i.e. addressing several scales of variation) of univariate or multivariate response data, reviewed, to our knowledge for the first time in this review. These methods are illustrated with ecological data from 13 years of benthic surveys in Chesapeake Bay, USA. The following methods are applied to the Chesapeake data: distance-based Moran's eigenvector maps, asymmetric eigenvector maps, scalogram, variation partitioning, multivariate correlogram, multivariate regression tree, and two-way MANOVA to study temporal and space-time variability. Local (temporal) contributions to beta diversity (LCBD indices) are computed and analysed graphically and by regression against environmental variables, and the role of species in determining the LCBD values is analysed by correlation analysis. A tutorial detailing the analyses in the R language is provided in an appendix.


Word-object and action-object association learning across early development.
Successful communication often involves comprehension of both spoken language and observed actions with and without objects. Even very young infants can learn associations between actions and objects as well as between words and objects. However, in daily life, children are usually confronted with both kinds of input simultaneously. Choosing the critical information to attend to in such situations might help children structure the input, and thereby, allow for successful learning. In the current study, we therefore, investigated the developmental time course of children's and adults' word and action learning when given the opportunity to learn both word-object and action-object associations for the same object. All participants went through a learning phase and a test phase. In the learning phase, they were presented with two novel objects which were associated with a distinct novel name (e.g., "Look, a Tanu") and a distinct novel action (e.g., moving up and down while tilting sideways). In the test phase, participants were presented with both objects on screen in a baseline phase, then either heard one of the two labels or saw one of the two actions in a prime phase, and then saw the two objects again on screen in a recognition phase. Throughout the trial, participants' target looking was recorded to investigate whether participants looked at the target object upon hearing its label or seeing its action, and thus, would show learning of the word-object and action-object associations. Growth curve analyses revealed that 12-month-olds showed modest learning of action-object associations, 36-month-olds learned word-object associations, and adults learned word-object and action-object associations. These results highlight how children attend to the different information types from the two modalities through which communication is addressed to them. Over time, with increased exposure to systematic word-object mappings, children attend less to action-object mappings, with the latter potentially being mediated by word-object learning even in adulthood. Thus, choosing between different kinds of input that may be more relevant in their rich environment encompassing different modalities might help learning at different points in development.


Effect of the broadcast of a television documentary about a teenager's suicide in Israel on suicidal behavior and methods.
Portrayals of suicide in the media are controversial because they may impact on suicide rates and methods. The aim of this study was to analyze the effect of the broadcast of a television documentary wherein an adolescent girl is interviewed about her suicide plan and subsequently dies by suicide. National suicide rates during the 8 weeks prior to the program and the 4 weeks following it were compared to the same periods in the previous year. There was no significant difference in the rates of completed and attempted suicides before and after the program. There was a shift toward the method used in the film after the broadcast, but it was found to be nonspecific to that year. A significant decrease was noted in the mean age of suicide attempters in the last promotional week prior to the broadcast compared to the previous year (p = .032). These preliminary findings suggest that the repeated televised promotion of a documentary on suicide may raise the risk of suicide in vulnerable populations. Further investigations in bigger populations are needed.


Barley consumption under a high-fat diet suppresses lipogenic genes through altered intestinal bile acid composition.
We evaluated whether barley flour consumption in a high-fat environment affects lipid metabolism through signals mediated by bile acids. Four-week-old mice were fed a high-fat diet supplemented with cellulose (HC) or β-glucan-rich barley flour (HB) for 12 weeks. Bile acid composition in the intestinal tract and feces was measured by GC/MS. Gene expression levels involved in bile acid metabolism in the liver and intestinal tract were determined by RT-PCR. Similar parameters were measured in mice treated with antibiotics (antibiotics-cellulose (AC) and antibiotics-barley (AB)) to reduce the activity of intestinal bacteria. The Results showed that the HB group had lower liver blood cholesterol and triglyceride levels than the HC group. The HB group showed a significant decrease in primary bile acids in the gastrointestinal tract compared to the HC group. On the other hand, the concentration of secondary bile acids relatively increased in the cecum and feces. In the liver, Fxr activation suppressed gene expression levels in synthesizing bile acids and lipids. Furthermore, in the gastrointestinal tract, Tgr5 was activated by increased secondary bile acids. Correspondingly, AMP levels were increased in the HB group compared to the HC group, AMPK was phosphorylated in the liver, and gene expression involved in lipid synthesis was downregulated. A comparison of the AC and AB groups treated with antibiotics did not confirm these effects of barley intake. In summary, our results suggest that the prevention of lipid accumulation by barley consumption involves signaling through changes in bile acid composition in the intestinal tract.


Genome-wide identification and expression analysis of the CPP-like gene family in soybean.
Cysteine-rich polycomb-like protein (CPP-like) genes are a group of transcription factors with highly conserved cysteine-rich domains and are widely distributed in animals and plants, but do not present in yeast. Previous studies have shown that members of this family play important roles in the development of reproductive tissue and in the control of cell division in plants. In this study, whole genome identification of soybean CPP transcription factors was performed using bioinformatic methods. The results showed that there were 20 CPP transcription factors in the soybean genome, which encoded for 28 distinct CPP proteins. These transcription factors were distributed on 16 of 20 chromosomes. Phylogenetic relationship analysis showed that expression of CPP gene family members occurred before the differentiation of monocotyledons and dicotyledons. RNA-Seq analysis showed that 5 genes were highly expressed in all tissues, including Glyma10g39080, Glyma01g44670, Glyma101g66920, Glyma02g01540, and Glyma20g28740. One gene (Glyma14g14750) was specifically expressed in young leaves, while 2 genes (Glyma02g01540 and Glyma10g01580) were highly expressed in root nodules. Quantitative reverse transcriptase-polymerase chain reaction analysis revealed that the expression levels of most genes increased in the roots under high temperature stress. Our findings indicate that these genes are not only involved in growth and development, but also in the responses to high temperature stress in soybean roots.


Molecular forms of prostate-specific antigen and human kallikrein 2 (hK2) in urine are not clinically useful for early detection and staging of prostate cancer.
Prostate-specific antigen (PSA), a member of the human kallikrein (hK) family, is the most important tumor marker for early detection, staging, and monitoring of men with prostate cancer today. However, the sensitivity of serum PSA is not sufficient to be used alone for prostate cancer screening. Recently, it was reported that the serum-to-urinary total PSA ratio improves the detection of men with prostate cancer, especially in men with a serum total PSA level between 4.0 and 10.0 ng/mL. We tested this hypothesis by evaluating the clinical usefulness of this PSA ratio as well as the use of the different molecular forms of PSA and human kallikrein 2 (hK2) in urine for detection and staging of prostate cancer. One hundred ten fresh, midstream urine specimens (prostate cancer 62, benign prostatic hyperplasia [BPH] 38, healthy male control 5, women 5) were collected. Serum total PSA, urine total PSA, urinary free PSA, urinary alpha 1-antichymotrypsin-bound PSA, and urinary hK2 levels were determined by monoclonal antibody assays (Hybritech Inc.). The serum-to-urinary total PSA ratio was calculated. The serum-to-urinary total PSA ratio did not accurately distinguish between men with BPH and men with prostate cancer. There was no significant difference between the urinary levels of any of the molecular forms of PSA or hK2 between men with prostate cancer and men with BPH. Among men with prostate cancer, neither urinary hK2 nor urinary levels of any of the molecular forms of PSA correlated with age, pathologic stage, or Gleason grade. In our study, the serum-to-urinary total PSA ratio did not improve the detection of men with prostate cancer. Furthermore, measurement of the molecular forms of PSA and hK2 in urine did not improve the detection or staging of prostate cancer over serum PSA alone.


Validation of FLOTAC for the detection and quantification of Troglodytella abrassarti and Neobalantidium coli in chimpanzees and pigs.
We carried out a calibration of FLOTAC for ciliates Troglodytella abrassarti and Neobalantidium coli based on the selection of a most appropriate flotation solutions, and we also tested its accuracy (i.e., number of detected stages out of known added number of stages to fecal samples) and sensitivity for trophozoites of both ciliates in chimpanzee feces and N. coli cysts in pig feces, compared the detection threshold of FLOTAC with MIF-based sedimentation, and, subsequently, tested the losses of ciliate stages during sample preparation. Nine flotation solutions were evaluated, and ZnSO4 solution (specific gravity [s.g.] 1.2) showed to be the most suitable for trophozoite detection, while Sheather's solution (s.g. 1.33) was selected as most suitable for cysts. The FLOTAC sensitivity in detection of both stages varied: for trophozoites, we found all samples were positive when the intensity of infection 10 trophozoites per gram and higher, whereas for cysts the sensitivity was lower. The accuracy of FLOTAC negatively correlated with infection intensity, and the merthiolate-iodine-formaldehyde sedimentation-based quantification had a lower detection threshold. We demonstrated additional losses of stages of T. abrassarti and N. coli due to their retention in the sediment, which is probably a major reason for discrepancies in the numbers of countable ciliates between both methods. In conclusion, the FLOTAC should not be considered as a gold standard for quantification of intestinal ciliates in primates; instead, we recommend the modified MIF method.


Changes in sperm quality and lipid composition during cryopreservation of boar semen.
The effect of cryopreservation on boar sperm viability, motility, lipid content and antioxidant enzymatic activities was studied. Three classes of semen were determined according to a cluster analysis on the basis of the proportion of live and dead cells after freezing and thawing. The classes identified were: high (H, n = 4), average (A, n = 12) and low (L, n = 3) viability. The concentration of sperm cells decreased from class H to A to L. Fresh semen samples with higher viability and a higher proportion of motile cells also maintained better quality after the freezing and thawing procedure. Sperm viability and motility in both fresh and thawed samples were similar in classes H and A, while significantly lower values were measured in class L. The relative decrease in sperm viability and motility after cryopreservation increased from class H to A to L. The lipid content of spermatozoa (micrograms per 10(9) cells) increased significantly after freezing and thawing in classes H and A but not in class L. This result indicated that active sperm lipid metabolism might be responsible for the increase in lipid content. Phospholipid and triacylglycerol contents increased whereas free cholesterol content decreased after thawing. The fatty acid composition of fresh spermatozoa was similar in all three classes. The proportion of polyunsaturated fatty acids decreased significantly after freezing and thawing, indicating contamination from the diluent or peroxidation. After freezing and thawing, superoxide dismutase activity in spermatozoa was significantly higher in class L than in classes H and A, which did not differ from each other.


The Effectiveness of Self-supervised Pre-training for Multi-modal Endometriosis Classification*†.
Endometriosis is a debilitating condition affecting 5% to 10% of the women worldwide, where early detection and treatment are the best tools to manage the condition. Early detection can be done via surgery, but multi-modal medical imaging is preferable given the simpler and faster process. However, imaging-based endometriosis diagnosis is challenging as 1) there are few capable clinicians; and 2) it is characterised by small lesions unconfined to a specific location. These two issues challenge the development of endometriosis classifiers as the training datasets tend to be small and contain difficult samples, which leads to overfitting. Hence, it is important to consider generalisation techniques to mitigate this problem, particularly self-supervised pre-training methods that have shown outstanding results in computer vision and natural language processing applications. The main goal of this paper is to study the effectiveness of modern self-supervised pre-training techniques to overcome the two issues mentioned above for the classification of endometriosis from multi-modal imaging data. We also introduce a new masking image modelling self-supervised pre-training method that works with 3D multi-modal medical imaging. Furthermore, to the best of our knowledge, this paper presents the first endometriosis classifier, fine-tuned from the pre-trained model above, which works with multi-modal (i.e., T1 and T2) magnetic resonance imaging (MRI) data. Our results show that self-supervised pre-training improves endometriosis classification by as much as 31%, when compared with classifiers trained from scratch.


Enhancing Generalization of Visuomotor Adaptation by Inducing Use-dependent Learning.
Learning a motor task in one condition typically generalizes to another, although it is unclear why it generalizes substantially in certain situations, but only partially in other situations (e.g., across movement directions and motor effectors). Here, we demonstrate that generalization of motor learning across directions and effectors can be enhanced substantially by inducing use-dependent learning, that is, by having subjects experience motor actions associated with a desired trajectory repeatedly during reaching movements. In Experiments 1 and 2, healthy human adults adapted to a visuomotor rotation while concurrently experiencing repetitive passive movements guided by a robot. This manipulation increased the extent of generalization across movement directions (Expt. 1) and across the arms (Expt. 2) by up to 50% and 42%, respectively, indicating crucial contribution of use-dependent learning to motor generalization. In Experiment 3, we applied repetitive transcranial magnetic stimulation (rTMS) to the left primary motor cortex (M1) of the human subjects prior to passive training with the right arm to increase cortical excitability. This intervention resulted in increased motor-evoked potentials (MEPs) and decreased short-interval intracortical inhibition (SICI) in the rTMS group, but not in the sham group. These changes observed in the rTMS group were accompanied by enhanced generalization of visuomotor adaptation across the arms, which was not the case in the sham group. Collectively, these findings confirm the involvement of M1 in use-dependent learning, and suggest that use-dependent learning can contribute not only to motor learning, but also to motor generalization.


The role of HB-donor groups in the heterocyclic polar fragment of H3-antagonists. I. Synthesis and biological assays.
It has been recently reported that compounds composed of an imidazole connected through an alkyl spacer to a 2-aminobenzimidazole showed high affinity towards the H(3)-receptor. The guanidine fragment of the 2-aminobenzimidazole is probably involved in hydrogen bond interactions at the binding site, and is referred to as the 'polar fragment'. In the present work, starting from 2-aminobenzimidazole derivatives with a di-methylene spacer 1 (pK(i)=7.25) or a tri-methylene one 2 (pK(i)=8.90), we investigated the importance of the hydrogen bond (HB) donor groups at the polar fragment in the interaction with the H(3)-receptor. The replacement of 2-aminobenzimidazoles with different moieties [2-aminobenzothiazole, 3, 4; 2-thiobenzimidazole, 5, 6; 2-thiobenzothiazole, 7, 8; 2-thio-4-phenyl- or 2-thio-5-phenyl-N-methylimidazoles, 9-12] highlighted the effect of the polar group basicity on the optimal length of the alkyl chain: longer spacers were preferred with polar groups of moderate basicity whereas, in the presence of neutral polar groups, the best affinity values were obtained with di-methylene chains. Moreover, N-methylation at the 2-aminobenzimidazole moiety 13-16 revealed different behaviour for compounds having different spacer lengths. In fact, methylation of the exocyclic NH group maintained high affinity for the tri-methylene 2-aminobenzimidazole derivative, while a drop in affinity was observed for the annular N-methylation. An opposite trend characterised di-methylene derivatives. These observed SAR suggest that, within this class of compounds, the number of HB-donor groups can be lowered while maintaining high receptor affinity. Since the presence of HB-donor groups strongly affects brain access, this observation could be useful to design and prepare new H(3)-antagonists.


Elevated ADAR expression is significantly linked to shorter overall survival and immune infiltration in patients with lung adenocarcinoma.
To date, few studies have investigated whether the RNA-editing enzymes adenosine deaminases acting on RNA (ADARs) influence RNA functioning in lung adenocarcinoma (LUAD). To investigate the role of ADAR in lung cancer, we leveraged the advantages of The Cancer Genome Atlas (TCGA) database, from which we obtained transcriptome data and clinical information from 539 patients with LUAD. First, we compared ARAR expression levels in LUAD tissues with those in normal lung tissues using paired and unpaired analyses. Next, we evaluated the influence of ADARs on multiple prognostic indicators, including overall survival at 1, 3 and 5 years, as well as disease-specific survival and progression-free interval, in patients with LUAD. We also used Kaplan-Meier survival curves to estimate overall survival and Cox regression analysis to assess covariates associated with prognosis. A nomogram was constructed to validate the impact of the ADARs and clinicopathological factors on patient survival probabilities. The volcano plot and heat map revealed the differentially expressed genes associated with ADARs in LUAD. Finally, we examined ADAR expression versus immune cell infiltration in LUAD using Spearman's analysis. Using the Gene Expression Profiling Interactive Analysis (GEPIA2) database, we identified the top 100 genes most significantly correlated with ADAR expression, constructed a protein-protein interaction network and performed a Gene Ontology/Kyoto Encyclopedia of Genes and Genomes analysis on these genes. Our results demonstrate that ADARs are overexpressed in LUAD and correlated with poor patient prognosis. ADARs markedly increase the infiltration of T central memory, T helper 2 and T helper cells, while reducing the infiltration of immature dendritic, dendritic and mast cells. Most immune response markers, including T cells, tumor-associated macrophages, T cell exhaustion, mast cells, macrophages, monocytes and dendritic cells, are closely correlated with ADAR expression in LUAD.


Fragile X protein controls neural stem cell proliferation in the Drosophila brain.
Fragile X syndrome (FXS) is the most common form of inherited mental retardation and is caused by the loss of function for Fragile X protein (FMRP), an RNA-binding protein thought to regulate synaptic plasticity by controlling the localization and translation of specific mRNAs. We have recently shown that FMRP is required to control the proliferation of the germline in Drosophila. To determine whether FMRP is also required for proliferation during brain development, we examined the distribution of cell cycle markers in dFmr1 brains compared with wild-type throughout larval development. Our results indicate that the loss of dFmr1 leads to a significant increase in the number of mitotic neuroblasts (NB) and BrdU incorporation in the brain, consistent with the notion that FMRP controls proliferation during neurogenesis. Developmental studies suggest that FMRP also inhibits neuroblast exit from quiescence in early larval brains, as indicated by misexpression of Cyclin E. Live imaging experiments indicate that by the third instar larval stage, the length of the cell cycle is unaffected, although more cells are found in S and G2/M in dFmr1 brains compared with wild-type. To determine the role of FMRP in neuroblast division and differentiation, we used Mosaic Analysis with a Repressible Marker (MARCM) approaches in the developing larval brain and found that single dFmr1 NB generate significantly more neurons than controls. Our results demonstrate that FMRP is required during brain development to control the exit from quiescence and proliferative capacity of NB as well as neuron production, which may provide insights into the autistic component of FXS.


The Effects of Molecular Hydrogen and Suberoylanilide Hydroxamic Acid on Paraquat-Induced Production of Reactive Oxygen Species and TNF-α in Macrophages.
The aim of this study is to investigate the effects of molecular hydrogen (H2) and suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, on paraquat (PQ)-stimulated production of reactive oxygen species (ROS) and tumor necrosis factor alpha (TNF-α) in macrophages. First, the PQ optimal concentration was determined in RAW264.7 macrophage by treating serum-starved cells with PQ at 0, 0.001, 0.01, 0.1, 1, and 10 mM. We evaluated at 1, 2 and 8 h (1) cell viability (by means of trypan blue exclusion method), (2) intracellular ROS levels (with a fluorescent DCFH-DA probe), and (3) TNF-α level in the culture media (determined by enzyme-linked immunosorbent assay, ELISA). Subsequently, mouse RAW267.4 macrophages were treated with PQ in combination with SAHA and/or H2 for 8 h. PQ exerted a significant stimulatory but nontoxic effect on RAW267.4 macrophages at 0.1 mM. This PQ concentration was used in the subsequent experiments. H2 and H2 combined with SAHA evoked a greater reduction in PQ-induced ROS production than SAHA alone, especially at 2 and 8 h. At 1 and 2 h, treatments involving H2 caused a greater decrease in PQ-induced production of TNF-α than the corresponding treatments without H2. However, at 8 h, treatment with SAHA evoked more pronounced effects on TNF-α than treatment without SAHA. H2 decreases PQ-induced ROS production and attenuates early PQ-induced TNF-α production whereas SAHA reduces the late phase of the PQ-induced TNF-α production in macrophages. The effects are enhanced by the combination of H2 and SAHA.


Differential arrival of leading and lagging strand DNA polymerases at fission yeast telomeres.
To maintain genomic integrity, telomeres must undergo switches from a protected state to an accessible state that allows telomerase recruitment. To better understand how telomere accessibility is regulated in fission yeast, we analysed cell cycle-dependent recruitment of telomere-specific proteins (telomerase Trt1, Taz1, Rap1, Pot1 and Stn1), DNA replication proteins (DNA polymerases, MCM, RPA), checkpoint protein Rad26 and DNA repair protein Nbs1 to telomeres. Quantitative chromatin immunoprecipitation studies revealed that MCM, Nbs1 and Stn1 could be recruited to telomeres in the absence of telomere replication in S-phase. In contrast, Trt1, Pot1, RPA and Rad26 failed to efficiently associate with telomeres unless telomeres are actively replicated. Unexpectedly, the leading strand DNA polymerase epsilon (Polepsilon) arrived at telomeres earlier than the lagging strand DNA polymerases alpha (Polalpha) and delta (Poldelta). Recruitment of RPA and Rad26 to telomeres matched arrival of DNA Polepsilon, whereas S-phase specific recruitment of Trt1, Pot1 and Stn1 matched arrival of DNA Polalpha. Thus, the conversion of telomere states involves an unanticipated intermediate step where lagging strand synthesis is delayed until telomerase is recruited.


Exogenous 1,25(OH)2D3/VD3 counteracts RSL3-Induced ferroptosis by enhancing antioxidant capacity and regulating iron ion transport: Using zebrafish as a model.
RSL3 is a common inhibitor of glutathione peroxidase 4 (GPx4) that can induce ferroptosis. Ferroptosis is an iron ion-dependent, oxidative-type of programmed cell death. In this study, larval/adult zebrafish were stimulated with RSL3 to construct a ferroptosis model, and CYP2R1-/- zebrafish was used as a 1,25(OH)2D3 knock-down model to explore the regulatory effect and mechanism of 1,25(OH)2D3/VD3 on RSL3-induced ferroptosis. The results showed that 1,25(OH)2D3/VD3 alleviated RSL3 induced mitochondrial damage in liver of larval/adult zebrafish, reversed the decline of GPx4 activity, and reduced the accumulation of ROS, LPO and MDA. VD3 also inhibited hepcidin (HEPC) in adult fish liver, promoted the production of ferroportin (FPN), and reduced the aggregation of Fe2+. Exogenous 1,25(OH)2D3 increased the CYP2R1-/- survival and liver GPx4 activity after RSL3 treatment. At the gene level, 1,25(OH)2D3/VD3 activated Keap1-Nrf2-GPx4 and inhibited the NFκB-hepcidin axis. In the ferroptosis context, deletion of the cyp2r1 gene resulted in a more severe decline in gpx4 expression, but the exogenous 1,25(OH)2D3 increased the expression of the GPx4 gene and protein in CYP2R1-/- zebrafish liver after RSL3 treatment. The collective results indicated that 1,25(OH)2D3/VD3 can inhibit ferroptosis induced by RSL3 in liver of larval/adult zebrafish by improving the antioxidant capacity and regulating iron ion transport. Exogenous 1,25(OH)2D3 reverses the downregulation of GPx4 in the CYP2R1-/- zebrafish liver in the ferroptosis state. Compared with the ferroptosis inhibitor Fer-1, the mechanism of action of 1,25(OH)2D3/VD3 is diversified and nonspecific. This study demonstrated the resistance of VD3 to RSL3-induced ferroptosis at different developmental stages in zebrafish.


Sensation seeking in a community sample of French gamblers: Comparison between strategic and non-strategic gamblers.
The purpose of this research is to examine the relationship between sensation seeking and gambling disorder (GD) in a community sample of gamblers (when controlling for the effect of substance use, gender and age) and see whether sensation seeking scores depend on the gambling activity when comparing strategic and non-strategic gamblers. A total of 380 gamblers was recruited. First, pathological gamblers (PGs) (n =143) were compared to non-pathological gamblers (NPGs) (n =237). Second, strategic gamblers (n =93) were compared to non-strategic gamblers (n =110). Sociodemographic data, gambling behavior (SOGS, DSM-IV), tobacco and alcohol use (CAGE), and sensation seeking (SSS) were evaluated. PGs have higher boredom susceptibility scores than NPGs and this factor is associated with GD. Nevertheless, the relationship between sensation seeking and GD depends on the gambling activity. In fact, sensation seeking is associated with GD in strategic gamblers only. PGs playing strategic games display different profiles from non-strategic PGs. Thus, factors associated with GD differ when the gambling activity is taken into account. These findings are consistent with the idea of it being essential to identify clinically distinct subgroups of PGs in the treatment of GD.


Haemoglobin A1 levels in Nigerian diabetic patients using microcolumn affinity chromatography.
Haemoglobin A1 (HbA1) levels were measured in forty-five diabetic Nigerians and thirteen non-diabetic controls using disposable microcolumn kits based on affinity chromatography. The effect on HbA1 results of storage of whole blood samples for 1 week at room temperature and in a refrigerator was also evaluated. The mean +/- s.d. of HbA1 levels in diabetic patients and controls were 12.1 +/- 5.6% and 5.2 +/- 0.90% respectively. The difference was highly significant, P less than 0.001. Newly diagnosed diabetic patients who had not been on any treatment had significantly higher mean value of HbA1, 14.4 +/- 5.0%, than old patients already on treatment, 9.5 +/- 2.9%, P less than 0.01. In a group of nine patients, the mean +/- s.d. of baseline HbA1 value was 10.3 +/- 4.2% and after 1 week storage of different portions of the samples at room temperature and in a refrigerator, the mean +/- s.d. respectively were 9.8 +/- 3.6% and 11.0 +/- 4.2%. There was thus a slight decrease on storage at room temperature and a slight increase during refrigeration but the observed differences were not statistically significant. It is concluded that the glyc-affinity micro-column chromatography is a satisfactory and potentially useful method for measuring HbA1 levels in tropical developing countries. The method was easy to use and the assays could be run under average ambient room temperature without recourse to use of an air-conditioned room or a special chromatography chamber.


Psychological mechanisms underlying support for juvenile sex offender registry laws: prototypes, moral outrage, and perceived threat.
In three studies, we investigated support for applying sex offender registry laws to juveniles. Family law attorneys supported registry laws less for juveniles than for adults. Laypeople and prosecutors supported juvenile and adult sex offender registration equally--even though they perceived juveniles as generally less threatening than adults (Study 1)--because most people spontaneously envision a severe sex offender prototype regardless of offender age (Study 2). People are less supportive of registry laws, however, when they envision less severe prototypes spontaneously (Study 2) or when induced to do so (Study 3). Effects of offender age, offender prototypes, and offense severity were mediated by perceptions of threat posed by the juvenile sex offender (i.e., utilitarian concerns). The effect of offense severity was also mediated by moral outrage (i.e., retributive concerns).


Electron Precise Sodium Carbaboride Nanocrystals from Molten Salts: Single Sources to Boron Carbides.
Boron-rich solids exhibit specific crystal structures and unique properties, which are only very scarcely addressed in nanoparticles. In this work, we address the original inorganic structural chemistry and reactivity of boron-rich nanoparticles, by reporting the first occurrence of sodium carbaboride nanocrystals based on the NaB5C crystal structure. To design these sub-10 nm nano-objects, we use liquid-phase synthesis in molten salts at 900 °C. By combining a set of characterization tools including powder X-ray powder diffraction, transmission electron microscopy, solid-state nuclear magnetic resonance coupled to DFT modeling, and X-ray photoelectron spectroscopy, we demonstrate that these nanocrystals deviate from the ideal stoichiometry reported for the bulk compound. We suggest that the carbon and sodium contents compensate each other to ensure that the octahedral cluster-based framework is stabilized by fulfilling an electron counting rule. These nanocrystals encompass substituted octahedral covalent structural building units not reported in the related bulk compound. They then shed new light on the ability of nanoparticles to host wide solid solution ranges in covalent solids and then to yield new solids. We finally show that these nanocrystals are efficient single sources of boron and carbon to form a nanostructured boron carbide, thus paving the way to new nanostructured materials.


Evaluation of the Sterifil lysis-filtration blood culture system.
This paper describes the comparison of the Sterifil lysis-filtration (SLF) blood culture procedure with a standard Trypticase soy broth (TSB) technique. The lysing solutions employed in the SLF system, Triton X-100 (alkyl phenoxy polyethoxy ethanol) and sodium carbonate, were deleterious to most bacteria commonly encountered in bacteremia except staphylococci and enterococci. Candida was not adversely affected. There was a positive correlation between the tolerance of the microbial isolants to the lysing solutions and their recovery by the SLF technique. A total of 3,554 cultures were run in parallel and 398 isolants were obtained. Of 201 gram-positive isolants, 135 were recovered by both techniques, 43 were detected by the TSB technique only, and 23 were recovered only with the SLF method. In sharp contrast, of 168 gram-negative isolants, 28 were recovered in common, 130 were isolated only by TSB, and 10 were recovered only with the SLF method. The SLF method detected all cases of candidemia detected by the TSB method plus an additional 12 for a total of 29 cases. The SLF method, as currently described, is generally too toxic to bacteria for routine use in a clinical laboratory.


Experimental surface strain mapping of porcine peripapillary sclera due to elevations of intraocular pressure.
To experimentally characterize 2D surface mapping of the deformation pattern of porcine peripapillary sclera following acute elevations of intraocular pressure (IOP) from 5 mm Hg to 45 mm Hg. Four porcine eyes were obtained within 48 h postmortem and dissected to the sclera. After the anterior chamber was removed, each posterior scleral shell was individually mounted at the equator on a custom-built pressurization device, which internally pressurized the scleral samples with isotonic saline at 22 degrees C. Black polystyrene microspheres (10 microm in diameter) were randomly scattered and attached to the scleral surface. IOP was incrementally increased from 5 mm Hg to 45 mm Hg (+/-0.15 mm Hg), and the surface deformation of the peripapillary sclera immediately adjacent to the dural insertion was optically tracked at a resolution of 2 micrompixel one quadrant at a time, for each of four quadrants (superior, nasal, inferior, and temporal). The 2D displacement data of the microsphere markers were extracted using the optical flow equation, smoothed by weighting function interpolation, and converted to the corresponding Lagrangian finite surface strain. In all four quadrants of each eye, the principal strain was highest and primarily circumferential immediately adjacent to the scleral canal. Average maximum Lagrangian strain across all quadrants for all eyes was 0.013+/-0.005 from 5 mm Hg to 10 mm Hg, 0.014+/-0.004 from 10 mm Hg to 30 mm Hg and 0.004+/-0.001 from 30 mm Hg to 45 mm Hg, demonstrating the nonlinearity in the IOP-strain relationship. For each scleral shell, the observed surface strain mapping implied that the scleral stiffness was relatively low between 5 mm Hg and 10 mm Hg, but dramatically increased for each IOP elevation increment beyond 10 mm Hg. Peripapillary deformation following an acute IOP elevation may be governed by the underlying scleral collagen microstructure and is likely in the high-stiffness region of the scleral stress-strain curve when IOP is above 10 mm Hg.


[Alcoholic mothers and their offspring].
The authors examined 75 mothers suffering from alcoholism with regard to risk factors for their progeny. When children had psychological pathology, the mothers' alcoholism was distinguished by greater severity of biological manifestations and relatively easier social sequelae. A peculiar nature of the alcoholism course, along with the alcohol doses taken by the mothers during pregnancy, determined the birth of children with the syndrome of alcoholic embryopathy. The appearance of children with neurotic and psychopathic disturbances reflects the polymorphism of the psychogenic effect of maternal alcoholism. A multifactorial effect of the mother's alcoholism on her progeny is postulated and practical recommendations based on this postulation are offered.


Endogenous Nitric Oxide Generation Linked to Ryanodine Receptors Activates Cyclic GMP / Protein Kinase G Pathway for Cell Proliferation of Neural Stem/Progenitor Cells Derived From Embryonic Hippocampus.
Nitric oxide (NO) activates the cyclic GMP (cGMP) / protein kinase G (PKG) pathway during physiological processes in numerous types of cells. Here, we evaluated whether this NO/cGMP/PKG pathway is involved in the proliferation of neural stem/progenitor cells (NPCs) derived from the hippocampus of embryonic mice. In culture, the exposure to the NO synthase inhibitor Nω-nitro-l-arginine methyl ester (l-NAME) significantly decreased the number of viable cells and 5-bromo-2'-deoxyuridine (BrdU) incorporation into the cells, as well as the levels of intracellular reactive oxygen species, extracellular NO2, and intracellular cGMP. Like l-NAME, the soluble guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) and PKG inhibitor KT5823 also decreased cell viability and BrdU incorporation. The membrane-permeable cGMP analogue 8-bromo-cGMP partially abolished the l-NAME-induced decrease in the BrdU incorporation. BrdU incorporation was decreased by Ca2+-channel blockers, including dantrolene, MK-801, ifenprodil, and nifedipine. Interestingly, the NO2 level was decreased by dantrolene, but not by the other 3 blockers. l-NAME and ODQ attenuated phosphorylation of Akt, but not that of extracellular signal-regulated kinases or epidermal growth factor receptors. Our data suggest that endogenous NO generation linked to dantrolene-sensitive ryanodine receptors activates the cGMP/PKG signaling pathway for positive regulation of proliferation of hippocampal NPCs derived from embryonic mice.


Antioxidant activities of fractions of polymeric procyanidins from stem bark of Acacia confusa.
The polymeric procyanidins extracted from Acacia confusa stem bark were fractionated with a step gradient of water, methanol and acetone on a Sephadex LH-20 column. The antioxidant activity of the collected fractions was investigated by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and ferric reducing/antioxidant power (FRAP) assays. All fractions possessed potent antioxidant activity with the highest activity observed for fraction F9. The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and reversed-phase high performance liquid chromatography (RP-HPLC) analyses suggested that the collected fractions consisted primarily of oligomeric and polymeric procyanidins, with different polymer ranges and most abundant polymer size. For each fraction, catechin and epicatechin were present as both terminal and extension units, and epicatechin was the major component in the extended chain. The mean degree of polymerization (mDP) of each fraction differed, ranging from 1.68 (fraction F2) to 17.31 (fraction F11). There was a relationship between antioxidant activity (IC50/DPPH and FRAP) and mDP (R(2) (DPPH) = 0.861, P = 0.006 and R(2) (FRAP) = 0.608, P = 0.038), respectively. However, the highest antioxidant activity of fraction (F9) was not coincident with the maximum mDP of fraction (F11).


More T Cells versus Better T Cells in Patients with Breast Cancer.
<b/> Characterization of intratumoral leukocyte populations may help in estimating patients' prognosis and predictions. A new study shows remarkable differences between the in situ and invasive breast cancer stages and highlights that cell number analyses should be complemented with characterization of T-cell functions, increasing the likelihood that immune competent antitumor T cells and further biomarkers are identified for guiding therapy choices. Cancer Discov; 7(10); 1062-4. ©2017 AACR.See related article by Gil Del Alcazar et al., p. 1098.


Involvement of substance P in peripheral neuropathy induced by paclitaxel but not oxaliplatin.
The painful peripheral neuropathy occurring frequently during chemotherapy with paclitaxel or oxaliplatin is one of their dose-limiting factors. We reported previously that substance P is involved in the pathogenesis of pulmonary hypersensitivity reaction to paclitaxel in rats, and an antiallergic agent pemirolast reverses this reaction via the blockade of release of substance P. In the present study, we investigated the involvement of substance P in paclitaxel-induced peripheral neuropathy compared with that by oxaliplatin. In von Frey and acetone tests in rats repeated administration of paclitaxel (6 mg/kg i.p., once a week for 4 weeks) or oxaliplatin (4 mg/kg i.p., twice a week for 4 weeks) induced both mechanical allodynia and cold hyperalgesia. Paclitaxel-induced peripheral neuropathy was reversed primarily by the acute administration of pemirolast (0.1 and 1 mg/kg p.o.). Moreover, coadministration of the receptor antagonists neurokinin 1 [N-acetyl-l-tryptophan 3,5-bis(trifluoromethyl)benzylester (L-732,138), 100 μg/body i.t.] and neurokinin 2 [5-fluoro-3-[2-[4-methoxy-4-[[(R)-phenylsulphinyl]methyl]-1-piperidinyl]ethyl]-1H-indole (GR159897), 100 μg/body i.t.] strongly reversed paclitaxel-induced neuropathy. On the other hand, oxaliplatin-induced peripheral neuropathy was not reversed by pemirolast. In the in vitro study using cultured adult rat dorsal root ganglion neurons paclitaxel (1000 ng/ml) significantly increased the release of substance P, and pemirolast (100 and 1000 nM) significantly inhibited this increase of substance P release. Oxaliplatin, by contrast, did not increase the release of substance P. These results suggest that substance P is involved in paclitaxel-induced neuropathy, and the mechanism of its action is clearly different from that of oxaliplatin.


Revisiting the single-stage forehead flap in nasal reconstruction.
To review the single-stage forehead flap for patient selection, technique, and main outcome measures. Patients undergoing nasal reconstruction between January 1, 1995, and June 30, 2000, were reviewed from medical records, photographs, and personal communication. All work was performed in an academic medical center. Fifty-one patients had a forehead flap for nasal reconstruction, of which 10 (20%) were repaired in a single stage. All patients had no evidence of small vessel disease, eg, hypertension, diabetes mellitus, or tobacco use. Nasal defects were limited to the upper two-thirds of the nose. The technique is modified from the original description by creating a unilateral, subcutaneous pedicle, wide undermining, and partial resection of the procerus muscle. One patient had superficial epidermolysis at the distal tip of the flap. The remaining 9 patients maintained complete viability with satisfactory outcomes. One debulking procedure was performed to the glabellar area for aesthetic reasons. The average interval for returning to work was 6.6 days compared with the minimal 3 weeks for conventional interpolated flaps. In select cases, a single-stage, island midline forehead flap can be used safely as an advantageous alternative to the conventional interpolated forehead flap.


[Modulation of Na+-K+-ATPase by calcium ions].
The modulatory effects of calcium ions on highly active Na+, K(+)-ATPase from calf brain and pig kidney tissues have been studied. The inhibitory action of Ca2+free on this enzyme depends on the level of ATP (but not AcP). The reduction of pH from 7.4 to 6.0 noticeably increases, but the elevation of pH to 8.0, in its turn, decreases the inhibition of ATP-hydrolyzing activity by calcium. With the increase of K+ concentration (in contrast to Na+) the sensibilization of Na+, K(+)-ATPase to Ca ions is observed. In the presence of potassium ions Mg2+free effectively modifies the inhibitory action of Ca2+free on this enzyme. Ca2+free (0.16-0.4 mM) decreases the sensitivity of Na+, K(+)-ATPase to action of the specific inhibitor ouabain in the presence of ATP. In the presence of AcP (phosphatase reaction) such a change of enzyme sensitivity to ouabain isn't observed. The influence of membranous effects of Ca2+ on the interaction of Na+, K(+)-ATPase with the essential ligands and cardiosteroids is discussed.


An experimental assessment of different anaerobic blood culture methods.
Simulated blood cultures were performed to test the ability of various methods to yield growth from small inocula of different strains of bacteroides and anaerobic cocci. For good early growth and reliable isolation of non-sporing anaerobes a Southern Group Brewers thioglycollate method, without Liquoid, appeared to be the best of the methods tested. A glucose cooked meat method also gave good results. Liquoid had no inhibitory effect on bacteroides. Anaerobic cocci were inhibited by Liquoid in some media, particularly thioglycollate, where one out of six strains tested was completely suppressed.


High hydrostatic pressure treatment of porcine oocytes induces parthenogenetic activation.
An innovative technique called high hydrostatic pressure (HHP) treatment has recently been reported to improve the cryosurvival of gametes and embryos in certain mammalian species, including the mouse, pig, and cattle. In the present study the parthenogenetic activation (PA) of pig oocytes caused by HHP treatment was investigated in different holding media with or without Ca(2+). The efficiency of activation was tested at different pressure levels and media including T2 (HEPES-buffered TCM-199 containing 2% cattle serum), and mannitol-PVA fusion medium with (MPVA + Ca(2+)) or without Ca(2+) and Mg(2+)(MPVA). The results showed that HHP cannot induce PA in T2, but only in MPVA + Ca(2+) with low Ca(2+) concentration and MPVA without Ca(2+). The highest activation efficiency was achieved with 10 min HHP treatment using 100 or 200 bars for oocytes in MPVA + Ca(2+) or MPVA, respectively. In the light of these results, the possible source of Ca(2+) during activation was investigated. It was found that even after a total of 30-min wash with TL-HEPES-PVA buffer without Ca(2+) before HHP treatment in MPVA, the oocytes could still be activated, indicating the possibility of an intracellular Ca(2+) source caused cytoplasmic free Ca(2+) elevation. In conclusion, parthenogenetic activation could be induced by HHP in certain holding media with low or zero Ca(2+) content. Further experiments are needed to identify the exact mechanisms of activation.


[Molecular mechanisms underlying bacillus anthracis infection at early stages and search for novel vaccines].
The developmental mechanisms of anthrax immunity were studied. Immunization was found to generally generate specific antibodies and lysozyme. Collectively, all the factors are responsible for suppressing the development of spores in the body. This proves the fact that the immunity is directed not only towards the exotoxin of B. anthracis, but it affects mainly the formation of vegetative cells. On entering the immuned body, vegetative cells may cause B. anthracis infection because antitoxic antibodies have no effect on encapsulated cells. The findings indicate that any anti-anthrax vaccine strain must show a complete immunological response in the body, as well as constitute immunity to all pathogenetic factors of B anthracis.


Differential kinetics of intracellular caspase-1-like and caspase-3-like enzyme activity in human alloreactive CD4(+) and CD8(+) T cells undergoing apoptosis.
The ICE-like families of serine proteases (caspases) have integral roles in apoptosis. These studies were performed to further understand the role of two critical caspases in relation to apoptotic regulation of the alloimmune response. A novel three-color cytofluorographic technique was utilized for measuring intracellular (in situ) caspase-1-like and caspase-3-like enzyme activity in responding CD4(+) and CD8(+) T cells over several time points of human mixed lymphocyte reactions (MLR). We found that activity levels of caspase 3 in both CD4(+) and CD8(+) responder cells began rising at day 10 of the MLR and peaked at day 14. By comparison, caspase 1 demonstrated the highest activity at day 7 in both cell subpopulations. These results coincided with the appearance of apoptotic cells among the alloreactive cells in the MLR. These findings demonstrate that intracellular caspase-1- and -3-like enzyme activity increases in both CD4(+) and CD8(+) alloreactive T cells as the primary response to allostimulatory cells progresses. While the kinetic profiles for these enzymes differed, both had a temporal association with the appearance of apoptosis in the MLR-generated cells. In all cases, the highest enzyme activity and presence of apoptosis was seen subsequent to the peak proliferative period. These results support the concept that changes in the rate and amount of apoptosis in alloreactive T cells is one mechanism by which the response to alloantigens is attenuated (i.e., tolerance) or sustained.


Reconstructionist molecular biology.
In the editorial inaugurating this journal, Levine (1989) pointed to a new reductionism in biology, which--unlike the old reductionism that led to specialization and isolation of areas concerned with different aspects of a complex biological problem--is providing a renewed sense of unity. This development is the result of widespread use of common experimental methodology and the emergence of signal transmission and differential gene expression as themes that are central to many areas of modern biology. I describe here a set of complementary developments in molecular biology that focus attention on the problems of complexity and organization. Simple examples are given that illustrate the difficulty of relating systemic behavior to the properties of the underlying molecular determinants, and the outlines of a general approach to this problem are presented. These developments, together with those highlighted by Levine, are leading us to a new, more integrative intellectual paradigm whose fruits will be the elucidation of fundamental issues concerning network function, design, and evolution that cannot be addressed by the current paradigm.


Enhanced angiotensin-mediated facilitation of adrenergic neurotransmission in spontaneously hypertensive rats.
The effect of angiotensin II on the pressor response of the mesenteric vascular bed to periarterial adrenergic nerve stimulation (PNS) was compared between spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). Angiotensin II (1-20 ng/ml) caused a marked potentiation of the vasoconstrictor response to PNS (8 Hz, 2 msec, for 30 sec) in a concentration-dependent manner without significantly affecting the base-line perfusion pressure in either WKY or SHR. The facilitatory effect of angiotensin II was significantly greater in SHR than in WKY preparations. The effective concentration for the peptide in SHR was approximately one-third of that found in WKY. Angiotensin II also potentiated the pressor response to exogenously administered norepinephrine (NE, 50 ng) in both WKY and SHR. The degree of potentiation of the response to PNS was greater than that to NE in SHR, while this differential was found only at high concentrations of angiotensin II on the WKY. The facilitatory effect of angiotensin II on the pressor response to either PNS or NE was markedly reduced by [Sar1-Ile8]angiotensin II (200 ng/ml) in both WKY and SHR. Angiotensin II caused further potentiation of the vasoconstrictor response to PNS or NE infusion in the presence of cocaine (5 micrograms/ml), which was of a greater magnitude in SHR than in WKY. These results suggest that the presynaptic facilitatory modulation of adrenergic vascular neurotransmission, mediated by angiotensin II receptors, is enhanced in the perfused mesenteric vascular bed of SHR.


Velocity measurements and flow patterns within the hinge region of a Medtronic Parallel bileaflet mechanical valve with clear housing.
During recent clinical trials the Medtronic Parallel bileaflet mechanical heart valve was found to have an unacceptable number of valves with thrombus formation when implanted in the mitral position. Thrombi were observed in the hinge region and also in the upstream portion of the valve housing in the vicinity of the hinge. It was hypothesized that the flow conditions inside the hinge may have contributed to the thrombus formation. In order to investigate the flow structures within the hinge, laser Doppler anemometry (LDA) measurements were conducted in both steady and pulsatile flow at approximately 70 predetermined sites within the hinge region of a 27 mm Medtronic Parallel mitral valve with transparent housing. The pulsatile flow velocity measurements were animated in time using a graphical software package to visualize the hinge flow field throughout the cardiac cycle. The LDA measurements revealed that mean forward flow velocities through the hinge region were on the order of 0.10-0.20 m/s. In the inflow channel, a large vortical structure was present during diastole. Upon valve closure, peak reverse velocity reached 3 m/s close to the housing wall in the inflow channel. This area also experienced high turbulent shear stresses (> 6000 dynes/cm2) during the leakage flow phase. A disturbed, vortical flow was again present in the inflow channel after valve closure, while slightly above the leaflet peg and relief the flow was essentially stagnant. The high turbulent stresses near the top of the inflow channel, combined with a persistent vortex, implicate the inflow channel of the hinge as a likely region of thrombus formation. This experimental investigation revealed zones of flow stagnation in the inflow region of the hinge throughout the cardiac cycle and elevated turbulent shear stress levels in the inflow region during the leakage flow phase. These fluid mechanic phenomena are most likely a direct result of the complex geometry of the hinge of this valve. Although the LDA measurements were conducted at only a limited number of sites within the hinge, these results suggest that the hinge design can significantly affect the washout capacity and thrombogenic potential of the Medtronic Parallel bileaflet mechanical heart valve. The use of LDA within the confines of the hinge region of a mechanical heart valve is a new application, made possible by recent advances in manufacturing technologies and a proprietary process developed by Medtronic that allowed the production of a transparent valve housing. Together, these modalities represent a new method by which future valve designs can be assessed before clinical trials are initiated.


Interactions of mitomycin C with mammalian DNA detected by alkaline elution.
The antitumor antibiotic mitomycin C (MMC) was studied in vitro using L1210 leukemia and 8226 human myeloma cells. Cytotoxicity was evaluated by colony formation in soft agar, and DNA damage was analyzed using alkaline elution filter assays. The purposes of these studies were: (a) to characterize the time course of MMC-DNA damage; (b) to characterize the type of DNA damage [DNA-DNA interstrand cross-links (ISC), DNA-protein cross-links (DPC), single and double strand breaks (SSBs, DSBs)]; and (c) to correlate this damage with cytotoxicity in vitro. Colony-forming assays showed the D0 value for 1 h MMC to be 15.0 microM for L1210 cells and 17 microM for 8226 cells. Alkaline elution studies showed that dose-dependent ISCs and DPCs formed rapidly following MMC exposure. Removal of cross-links was delayed, with only 50% repaired 32 h after exposure. There was a good correlation between ISCs and cytotoxicity in dose-response studies in each cell line. ISCs appeared to comprise most of the MMC-DNA lesions in both cell lines. No DNA SSBs or DSBs were observed following MMC exposure. Nuclei isolated from both cell lines and exposed to MMC produced less MMC alkylation than whole cells but, again, no strand breaks were evident. These results demonstrate that MMC is principally an alkylating agent when used at pharmacological (cytotoxic) concentrations in vitro. The lack of evidence for DNA strand breaks discounts a significant role for putative quinone-generated oxygen free radicals in the production of MMC cytotoxicity.


Calcium-dependent dimerization of human soluble calcium activated nucleotidase: characterization of the dimer interface.
Mammals express a protein homologous to soluble nucleotidases used by blood-sucking insects to inhibit host blood clotting. These vertebrate nucleotidases may play a role in protein glycosylation. The activity of this enzyme family is strictly dependent on calcium, which induces a conformational change in the secreted, soluble human nucleotidase. The crystal structure of this human enzyme was recently solved; however, the mechanism of calcium activation and the basis for the calcium-induced changes remain unclear. In this study, using analytical ultracentrifugation and chemical cross-linking, we show that calcium or strontium induce noncovalent dimerization of the soluble human enzyme. The location and nature of the dimer interface was elucidated using a combination of site-directed mutagenesis and chemical cross-linking, coupled with crystallographic analyses. Replacement of Ile(170), Ser(172), and Ser(226) with cysteine residues resulted in calcium-dependent, sulfhydryl-specific intermolecular cross-linking, which was not observed after cysteine introduction at other surface locations. Analysis of a super-active mutant, E130Y, revealed that this mutant dimerized more readily than the wild-type enzyme. The crystal structure of the E130Y mutant revealed that the mutated residue is found in the dimer interface. In addition, expression of the full-length nucleotidase revealed that this membrane-bound form can also dimerize and that these dimers are stabilized by spontaneous oxidative cross-linking of Cys(30), located between the single transmembrane helix and the start of the soluble sequence. Thus, calcium-mediated dimerization may also represent a mechanism for regulation of the activity of this nucleotidase in the physiological setting of the endoplasmic reticulum or Golgi.


The role of polysaccharide peptide of Ganoderma lucidum as a potent antioxidant against atherosclerosis in high risk and stable angina patients.
Antioxidants can reduce oxidative radicals that affect the early phase of atherogenesis, that is endothelial dysfunction. Polysaccharide Peptide (PsP) derived from Ganoderma lucidum has an active substance in the form of β-glucan. Previous studies have proven the PsP of Ganoderma lucidum as an effective antioxidant in atherosclerotic rats and shows no toxicity in animal model. This study aims to prove the effect of PsP as potent antioxidant in high risk and stable angina patients. This is a clinical trial conducted to 37 high risk and 34 stable angina patients, which were determined based on ESC Stable CAD Guidelines and Framingham risk score, with pre and post test design without control group. The parameters are superoxide dimustase (SOD) and malondialdehyde (MDA) concentration, circulating endothelial cell (CEC) and endothelial progenitor cell (EPC) counts. The patients were given PsP 750mg/day in 3 divided dose for 90days. Paired t-test was performed for normally distributed data, and Wilcoxon test for not normally distributed data, and significant level of p≤0,05. SOD level in high risk patients slightly increased but not statistically significant with p=0,22. Level of SOD in stable angina group significantly increased with p=0,001. MDA concentration significantly reduced in high risk and stable angina patients with p=0.000. CEC significantly reduced both in high risk and stable angina patients, with p=0.000 in both groups. EPC count significantly reduced in high risk and stable angina with p=0.000. PsP of Ganoderma lucidum is a potent antioxidant against pathogenesis of atherosclerosis in stable angina and high risk patients.


Identifying high-risk individuals for cardiovascular disease: similarities between venous and arterial thrombosis in perspective. A 2011 update.
The aim of this narrative review is to assess the potential association between arterial and venous thrombotic events. Several studies have suggested that the major cardiovascular risk factors, alone or in combination (e.g. in the metabolic syndrome), are significantly associated with venous thromboembolism (VTE). Recent evidence also suggests that microalbuminuria and non-alcoholic liver steatosis, both markers of arterial disease, may independently predict the risk for VTE. An association between a history of VTE and the risk of future arterial events is also well documented, inflammation and endothelial dysfunction being thought as the common soil on which further investigation in the area should be pursued. The existence of a common pathophysiologic background is also suggested by the evidence that aspirin, low-molecular weight heparin (LMWH) and warfarin are recommended for the prevention and treatment of both venous and arterial thrombosis. In addition, rosuvastatin recently has been shown to prevent venous thromboembolism (VTE) in a time-dependent fashion. Together, these data argue for patients with a history of VTE as being at intermediate/high cardiovascular risk, a concept that implies that VTE patients should undergo a careful assessment for the presence of cardiovascular risk factors and adequate lifestyle changes. The value of routine screening for asymptomatic atherosclerosis (e.g. 2D echocardiography, microalbuminuria, arterial vessel ultrasonography) in these patients should be confirmed in future studies.


Exercise Science Graduates in the Healthcare System: A Comparison Between Australia and Switzerland.
Physical inactivity (PI) is a leading risk factor for global mortality worldwide, a major preventable cause of non-communicable diseases (NCDs) and a socioeconomic burden for healthcare systems. Fortunately, evidence shows that exercise interventions delivered by qualified exercise science graduates is an effective way to reduce PI, prevent and treat NCDs. This study compares the integration of exercise science graduates, defined as university graduates with degrees in sport and exercise science, in the healthcare systems of Australia, a commonly cited model in this regard, and Switzerland, a country considered to have an effective but costly healthcare system. For both countries, three domains were reviewed: healthcare system, exercise science graduates' education, and roles played by exercise science graduates in healthcare system. Australia formally recognizes specifically trained exercise science graduates (referred to as Accredited Exercise Physiologists) as healthcare professionals. The exercise interventions they deliver, which were shown to be cost-effective and lead to positive health outcomes, are covered by Medicare, the Australian universal health insurance. However, Medicare covers only a maximum of 5 yearly sessions of all allied-health services taken together. Conversely, Switzerland, despite offering university master's degrees that focus on physical activity delivery to clinical populations, does not recognize the respective graduates as healthcare providers. As a result, their services are not covered by the Swiss health insurances. The latter do, however, cover a generous number of services (not formally limited) delivered by other allied-health professionals. In conclusion, Australia makes a better use of exercise science graduates than Switzerland does. Switzerland would benefit from establishing a clinical profession for exercise science graduates, defining competencies that they should acquire and setting their scope of practice. The very restricted number of therapy sessions covered by Medicare might limit the positive impact exercise science graduates have on the Australian healthcare system. Overall, mutual learning between countries can promote development and global recognition of clinical positions for exercise science graduates.


Correction propagation for user-assisted optical coherence tomography segmentation: general framework and application to Bruch's membrane segmentation.
Optical coherence tomography (OCT) is a commonly used ophthalmic imaging modality. While OCT has traditionally been viewed cross-sectionally (i.e., as a sequence of B-scans), higher A-scan rates have increased interest in en face OCT visualization and analysis. The recent clinical introduction of OCT angiography (OCTA) has further spurred this interest, with chorioretinal OCTA being predominantly displayed via en face projections. Although en face visualization and quantitation are natural for many retinal features (e.g., drusen and vasculature), it requires segmentation. Because manual segmentation of volumetric OCT data is prohibitively laborious in many settings, there has been significant research and commercial interest in developing automatic segmentation algorithms. While these algorithms have achieved impressive results, the variability of image qualities and the variety of ocular pathologies cause even the most robust automatic segmentation algorithms to err. In this study, we develop a user-assisted segmentation approach, complementary to fully-automatic methods, wherein correction propagation is used to reduce the burden of manually correcting automatic segmentations. The approach is evaluated for Bruch's membrane segmentation in eyes with advanced age-related macular degeneration.