Pulmonary tissue resistance to Candida albicans in normal and in immunosuppressed mice.
We characterized the clearance of Candida albicans from the lung using a murine model for pulmonary aspiration. Swiss Webster mice uniformly survived intratracheally administered boluses of C. albicans (1 to 30 X 10(5) colony-forming units of yeast) which killed the majority of mice (more than 85%) when injected intravenously. Clearance studies, using quantitative cultures of lung homogenates, demonstrated rapid elimination of C. albicans from the lung after a 6-h delay; the residual fractions of viable fungi were 8.3 and 0.7% of the initial inoculums at 24 and 48 h, respectively, after inoculation. The number of leukocytes in the bronchoalveolar spaces increased twofold to threefold after deposition, and this primarily reflected a neutrophil influx. Histologic studies supported the bronchoalveolar lavage results and revealed a diffuse interstitial neutrophilic infiltrate and clusters of inflammatory cells in air spaces at 6 and 24 h after Candida deposition. Examination of lavage pellets demonstrated that both neutrophils and macrophages ingested C. albicans in vivo. Immunosuppression (orally administered prednisolone for 2 wk) delayed the clearance of C. albicans from the lung. However, evaluation of neutrophil migration into bronchoalveolar spaces and of the in vivo ingestion capacity of both macrophages and neutrophils did not identify differences that could explain this delayed clearance in steroid-treated mice. The fungicidal activity of pulmonary leukocytes was measured with in vitro assays and was similar in phagocyte cultures from control and steroid-treated mice. In summary, intrinsic pulmonary defense factors and recruited neutrophils rapidly and completely clear C. albicans from the lung after bolus deposition.(ABSTRACT TRUNCATED AT 250 WORDS)


Immune and Inflammatory Cell Composition of Human Lung Cancer Stroma.
Recent studies indicate that the abnormal microenvironment of tumors may play a critical role in carcinogenesis, including lung cancer. We comprehensively assessed the number of stromal cells, especially immune/inflammatory cells, in lung cancer and evaluated their infiltration in cancers of different stages, types and metastatic characteristics potential. Immunohistochemical analysis of lung cancer tissue arrays containing normal and lung cancer sections was performed. This analysis was combined with cyto-/histomorphological assessment and quantification of cells to classify/subclassify tumors accurately and to perform a high throughput analysis of stromal cell composition in different types of lung cancer. In human lung cancer sections we observed a significant elevation/infiltration of total-T lymphocytes (CD3+), cytotoxic-T cells (CD8+), T-helper cells (CD4+), B cells (CD20+), macrophages (CD68+), mast cells (CD117+), mononuclear cells (CD11c+), plasma cells, activated-T cells (MUM1+), B cells, myeloid cells (PD1+) and neutrophilic granulocytes (myeloperoxidase+) compared with healthy donor specimens. We observed all of these immune cell markers in different types of lung cancers including squamous cell carcinoma, adenocarcinoma, adenosquamous cell carcinoma, small cell carcinoma, papillary adenocarcinoma, metastatic adenocarcinoma, and bronchioloalveolar carcinoma. The numbers of all tumor-associated immune cells (except MUM1+ cells) in stage III cancer specimens was significantly greater than those in stage I samples. We observed substantial stage-dependent immune cell infiltration in human lung tumors suggesting that the tumor microenvironment plays a critical role during lung carcinogenesis. Strategies for therapeutic interference with lung cancer microenvironment should consider the complexity of its immune cell composition.


Distribution of natural antibody against human neuroblastoma among children with or without neuroblastoma.
This study was conducted to examine the distribution of a natural antibody against neuroblastoma (NB) among Japanese children and to clarify the clinical significance of the presence of this antibody in the sera during treatment in patients with International Neuroblastoma Staging System Stage 4 NB. Human sera were obtained from 8 healthy volunteers, 82 patients with non-malignant surgical diseases, and 35 patients with NB including 3 with Stage 1 disease, 6 with Stage 2, 7 with Stage 3, 17 with Stage 4, and 2 with Stage 4S. This natural antibody was quantified by flow cytometry and its anti-tumor activity was measured by complement-dependent cytotoxicity (CDC) using TGW cells, a human NB cell line, as the target. IgM antibody and CDC activity against NB could be detectetd in all sera from healthy volunteers and from patients with non-malignant surgical dis eases who were age >1 year. The amount of IgM antibody and CDC activity in sera from patients with Stage 4 NB at diagnosis consistently was low, most likely because of massive absorption by tumor cells. In this group of patients, the increased CDC activity detected during treatment was indicative of a favorable factor for survival. A natural antibody against NB appears to exist in the sera of Japanese children. The sequential assessment of the levels of this antibody in the sera from Stage 4 NB patients during treatment may serve as a prognostic indicator.


Minor environmental concentrations of antibiotics can modify bacterial virulence in co-infection with a non-targeted parasite.
Leakage of medical residues into the environment can significantly impact natural communities. For example, antibiotic contamination from agriculture and aquaculture can directly influence targeted pathogens, but also other non-targeted taxa of commensals and parasites that regularly co-occur and co-infect the same host. Consequently, antibiotics could significantly alter interspecific interactions and epidemiology of the co-infecting parasite community. We studied how minor environmental concentrations of antibiotic affects the co-infection of two parasites, the bacterium Flavobacterium columnare and the fluke Diplostomum pseudospathaceum, in their fish host. We found that antibiotic in feed, and particularly the minute concentration in water, significantly decreased bacterial virulence and changed the infection success of the flukes. These effects depended on the level of antibiotic resistance of the bacterial strains. Antibiotic, however, did not compensate for the higher virulence of co-infections. Our results demonstrate that even very low environmental concentrations of antibiotic can influence ecology and epidemiology of diseases in co-infection with non-targeted parasites. Leakage of antibiotics into the environment may thus have more complex effects on disease ecology than previously anticipated.


Nonlinear measures of heart rate variability and mortality risk in hemodialysis patients.
Nonlinear measures of heart rate variability (HRV) have gained recent interest as powerful risk predictors in various clinical settings. This study examined whether they improve risk stratification in hemodialysis patients. To assess heart rate turbulence, deceleration capacity, fractal scaling exponent (α(1)), and other conventional HRV measures, 281 hemodialysis patients underwent 24-hour electrocardiography between January 2002 and May 2004 and were subsequently followed up. During a median 87-month follow-up, 77 patients (27%) died. Age, left ventricular ejection fraction, serum albumin, C-reactive protein, and calcium × phosphate independently predicted mortality. Whereas all nonlinear HRV measures predicted mortality, only decreased scaling exponent α(1) remained significant after adjusting for clinical risk factors (hazard ratio per a 0.25 decrement, 1.46; 95% confidence interval [95% CI], 1.16-1.85). The inclusion of α(1) into a prediction model composed of clinical risk factors increased the C statistic from 0.84 to 0.87 (P=0.03), with 50.8% (95% CI, 20.2-83.7) continuous net reclassification improvement for 5-year mortality. The predictive power of α(1) showed an interaction with age (P=0.02) and was particularly strong in patients aged <70 years (n=208; hazard ratio, 1.87; 95% CI, 1.38-2.53), among whom α(1) increased the C statistic from 0.85 to 0.89 (P=0.01), with a 93.1% (95% CI, 59.3-142.0) continuous net reclassification improvement. Scaling exponent α(1) that reflects fractal organization of short-term HRV improves risk stratification for mortality when added to the prediction model by conventional risk factors in hemodialysis patients, particularly those aged <70 years.


Spotted fever rickettsioses. Study of cases reported in a secondary care pediatric hospital of northeastern Mexico, 2012-2022.
Spotted fever rickettsiosis is a serious disease with a high mortality rate if not timely detected. To describe the characteristics of patients hospitalized for spotted fever rickettsiosis, as well as the risk factors associated with poor prognosis. Data from medical records of patients hospitalized between August 2012 and July 2022 were collected. Variables were analyzed using Mann-Whitney's U-test, Fisher's exact test, and univariate or multivariate logistic regression analysis. Twenty-six patients were analyzed, among which a mortality of 57.6% was identified. In the between-group comparison, platelet count was lower in non-survivors (16.0 × 103/μL vs. 25.9 × 103/μL, p = 0.031). The percentage of surviving patients who received treatment more than 72 hours after fever onset was 45.5% (five patients) vs. 86.7% of non-survivors (13 patients) (p = 0.034). Receiving treatment 72 hours after fever onset increased by 7.09 times the probability of a fatal outcome (OR = 8.09, 95% CI = 1.1-55.8, p = 0.034). Starting adequate treatment 72 hours after the onset of fever may be an important risk factor for mortality, hence the importance of timely diagnosis and appropriate treatment of this disease.


Pyridine nucleotide cycle of Salmonella typhimurium: in vitro demonstration of nicotinamide mononucleotide deamidase and characterization of pnuA mutants defective in nicotinamide mononucleotide transport.
The enzyme nicotinamide mononucleotide deamidase, an integral component of the proposed four-membered pyridine nucleotide cycle (PNC IV), has been demonstrated in extracts of Salmonella typhimurium LT2. The enzyme has an optimum pH of 8.7 and deamidates nicotinamide mononucleotide, forming nicotinic acid mononucleotide. Sigmoidal kinetic data suggest that this enzyme may be allosteric and therefore an important regulatory component of pyridine nucleotide cycle metabolism. Mutants previously designated pncC in anticipation of their lacking nicotinamide mononucleotide deamidase were examined and found to have normal levels of this enzyme. [14C]nicotinamide mononucleotide uptake studies, however, revealed a defect in the transport of this compound. Accordingly, the genetic designation for this locus was changed to pnuA to reflect its involvement in pyridine nucleotide uptake. Evidence is presented for the existence of two separate nicotinamide mononucleotide transport systems.


Diagnostic Value of Transluminal Attenuation Gradient for the Presence of Ischemia as Defined by Fractional Flow Reserve and Quantitative Positron Emission Tomography.
The aim of this study was to investigate the incremental diagnostic value of transluminal attenuation gradient (TAG), TAG with corrected contrast opacification (TAG-CCO), and transluminal diameter gradient (TDG) over coronary computed tomography angiography (CTA)-derived diameter stenosis alone for the identification of ischemia as defined by both the invasive reference standard fractional flow reserve (FFR) and the noninvasive reference standard quantitative positron emission tomography (PET). In addition to anatomic information obtained by coronary CTA, several functional CT parameters have been proposed to identify hemodynamically significant lesions more accurately, such as TAG, TAG-CCO, and more recently TDG. However, clinical validation studies have reported conflicting results, and a recent study has suggested that TAG may be affected by changes in vessel diameter. Patients with suspected coronary artery disease underwent coronary CTA and [15O]H2O PET followed by invasive coronary angiography with FFR of all major coronary arteries. TAG, TAG-CCO, and TDG were assessed, and the incremental diagnostic value of these parameters over coronary CTA-derived diameter stenosis alone for ischemia as defined by PET (hyperemic myocardial blood flow ≤2.30 ml/min/g) and FFR (≤0.80) was determined. A total of 557 (91.9%) coronary arteries of 201 patients were included for analysis. TAG, TAG-CCO, and TDG did not discriminate between vessels with or without ischemia as defined by either PET or FFR. Furthermore, these parameters did not have incremental diagnostic accuracy over coronary CTA alone for the presence of ischemia as defined by PET and FFR. There was a significant correlation between TDG and TAG (r = 0.47; p < 0.001) and between TDG and TAG-CCO (r = 0.37; p < 0.001). TAG, TAG-CCO, and TDG do not provide incremental diagnostic value over coronary CTA alone for the presence of ischemia as defined by [15O]H2O PET and/or FFR. The lack of diagnostic value of contrast enhancement-based flow estimations appears related to coronary luminal dimension variability.


Dried tomato pomace supplementation to reduce lamb concentrate intake: Effects on growth performance and meat quality.
The effect of dried tomato pomace (DTP) was tested on lamb performances, meat fatty acids and oxidative stability. For 36 days, one group of lambs (n = 7; CON) was fed a commercial concentrate, while the other group (n = 7; DTP) received DTP in addition to CON diet. The administration of DTP reduced (P < .05) concentrate intake, with no effect on animal performances. The DTP treatment tended to increase total polyunsaturated fatty acids (PUFA; P = .075), PUFA n-6 (P = .071), α-linolenic acid (P = .096) and increased linoleic acid (P < .05), γ-tocopherol (P < .001) and retinol (P < .001) in meat. In raw meat, DTP treatment increased L* (P = .059), b* (P < .05), C* (P = .052) and H* (P < .05) values compared to CON, while lipid oxidation was not affected. In meat homogenates incubated with pro-oxidants, DTP tended to reduce 2-thiobarbituric acid reactive substances (TBARS; P = .088). Therefore, DTP supplementation decreased the consumption of commercial concentrate without detrimental effects on animal performances and meat quality traits.


Comparative evaluation of two commercial assays for direct detection of Mycobacterium tuberculosis in respiratory specimens.
Two commercial systems for the amplification and detection of Mycobacterium tuberculosis directly from respiratory samples were compared. The Roche Cobas Amplicor MTB Test and the Roche manual Amplicor MTB Test (Roche Diagnostic Systems, USA) were applied to 755 decontaminated respiratory specimens collected from 470 patients. Results were compared with those of acid-fast staining and culture. A total of 251 specimens were collected from 156 patients diagnosed with pulmonary tuberculosis, including 28 specimens corresponding to 13 patients that were receiving antituberculous treatment. Given the overall positivity rate of 33.2% (251/755), the sensitivity, specificity, and positive and negative predictive values were 92.4, 100, 100, and 96.5%, respectively, for the Cobas Amplicor MTB Test and 90.8, 100, 100, and 95.8%, respectively, for the Amplicor MTB Test. For 204 (81.3%) smear positive specimens and 47 (19.7%) smear negative specimens, the sensitivity values were 100 and 59.6%, respectively, for the Cobas Amplicor MTB Test and 100 and 51%, respectively, for the Amplicor MTB Test. There were no statistically significant differences in sensitivity or specificity between the two assays and culture (p>0.05). The overall results of both assays were concordant for 99.5% of the samples. It is concluded that although both nucleic acid amplification methods are rapid and specific for the detection of Mycobacterium tuberculosis complex in respiratory specimens, the Cobas Amplicor MTB Test appears to be slightly more sensitive than the Amplicor MTB Test when smear negative specimens are investigated.


Imputing Longitudinal Growth Data in International Pediatric Studies: Does CDC Reference Suffice?
This study investigates a missing value imputation approach for longitudinal growth data in pediatric studies from multiple countries. We analyzed a combined cohort from five natural history studies of type 1 diabetes (T1D) in the US and EU with longitudinal growth measurements for 23,201 subjects. We developed a multiple imputation methodology using LMS parameters of CDC reference data. We measured imputation errors on both combined and individual cohorts using mean absolute percentage error (MAPE) and normalized root-mean-square error (NRMSE). Our results show low imputation errors using CDC reference. Overall height imputation errors were lower than for weight. The largest MAPE for weight and height among all age groups was 4.8% and 1.7%, respectively. When comparing performance between CDC reference and country-specific growth charts, we found no significant differences for height (CDC vs. German: p =0.993, CDC vs. Swedish: p=0.368) and for weight (CDC vs. Swedish: p=0.513) for all ages.


SF3B1 homeostasis is critical for survival and therapeutic response in T cell leukemia.
The production of noncanonical mRNA transcripts is associated with cell transformation. Driven by our previous findings on the sensitivity of T cell acute lymphoblastic leukemia (T-ALL) cells to SF3B1 inhibitors, we identified that SF3B1 inhibition blocks T-ALL growth in vivo with no notable associated toxicity. We also revealed protein stabilization of the U2 complex component SF3B1 via deubiquitination. Our studies showed that SF3B1 inhibition perturbs exon skipping, leading to nonsense-mediated decay and diminished levels of DNA damage response-related transcripts, such as the serine/threonine kinase CHEK2, and impaired DNA damage response. We also identified that SF3B1 inhibition leads to a general decrease in R-loop formation. We further demonstrate that clinically used SF3B1 inhibitors synergize with CHEK2 inhibitors and chemotherapeutic drugs to block leukemia growth. Our study provides the proof of principle for posttranslational regulation of splicing components and associated roles and therapeutic implications for the U2 complex in T cell leukemia.


Regulation of the postburn wound inflammatory response by gammadelta T-cells.
Healing of the burn injury site is a critical component of the patient's successful recovery from this form of trauma. Previous studies from our laboratory have demonstrated that gammadelta T-cells via the production of growth factors are important in burn wound healing. Nonetheless, the role of these cells in burn wound inflammation remains unknown. To study this, wild-type (WT) and gammadelta T-cell receptor-deficient (delta TCR) C57BL/6 male mice were subjected to burn injury or sham procedure. Wound cells were collected by implantation of polyvinyl alcohol sponges beneath the burn site in injured mice or beneath uninjured skin in sham mice. At 3 days after injury, infiltrating cells, wound fluid, and skin were collected for analysis. Burn injury markedly increased skin tumor necrosis factor-alpha (TNF-alpha) and monocyte chemoattractant protein 1 levels. In WT mice, the numbers of infiltrating cells were similar between nonburn wounds and burn wounds. In contrast, deltaTCRmice displayed a 6-fold reduction in the cellular infiltrate. Burn injury in WT mice caused a marked increase in burn wound TNF-alpha, monocyte chemoattractant protein 1, and interleukin 6 content as compared with nonburn wounds, whereas in delta TCRmice, the burn-induced increase of TNF-alpha and interleukin 6 was not observed. The wound cell infiltrate at 3 days postinjury was devoid of gammadelta T-cells in WT mice. It was predominately of myeloid origin expressing high levels of CD11b and F4/80. In conclusion, these findings suggest that resident gammadelta T-cells are important in the recruitment of inflammatory cells and regulation of the inflammatory response at the wound site after thermal injury.


Bisphenol A, phthalate metabolites and glucose homeostasis in healthy normal-weight children.
Bisphenol A and several of the most commonly used phthalates have been associated with adverse metabolic health effects such as obesity and diabetes. Therefore, we analyzed these man-made chemicals in first morning urine samples from 107 healthy normal-weight Danish children and adolescents. This was a cross-sectional study. Participants were recruited as part of the Copenhagen Puberty Study. The subjects were evaluated by an oral glucose tolerance test (OGTT), a dual-energy X-ray absorptiometry (DXA) scan, direct oxygen uptake measurement during cycle ergometry and fasting blood samples. First morning urine was collected and phthalate metabolites and BPA were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with prior enzymatic deconjugation. Individual chemical concentrations were divided into tertiles and analyzed in relation to biological outcome. Children in the lowest tertile of urinary BPA had significantly higher peak insulin levels during OGTT (P = 0.01), lower insulin sensitivity index (P < 0.01), higher leptin (P = 0.03), triglyceride (P < 0.01) and total cholesterol levels (P = 0.04), lower aerobic fitness (P = 0.02) and a tendency toward higher fat mass index (P = 0.1) compared with children in the highest tertile for uBPA. No significant differences in anthropometrics, body composition or glucose metabolism were associated with any of the phthalate metabolites measured. This pilot study on healthy normal-weight children suggests an inverse association between BPA and insulin resistance. Our findings contrast other cross-sectional studies showing a positive association for BPA, which may be due to confounding or reverse causation because diet is an important source of both BPA exposure and obesity.


Length effects in VUV photofragmentation of protonated peptides.
We have studied photoionization of protonated synthetic peptides YG(n)F (n = 0, 1, 3, 5, 10). Photon energies ranging from 8 to 30 eV were used. For YG(n)F peptides up to n = 5 small fragment ions related to the sidechains of the aromatic terminal amino acids Y and F dominate the fragmentation patterns. The associated yields scale with total photoabsorption cross section, demonstrating efficient hole migration towards the terminal amino acids upon photoionization of the peptide backbone. For n = 10 the side-chain loss channel is quenched and a series of large dications appear.


HIV Care Continuum Disparities Among Black Bisexual Men and the Mediating Effect of Psychosocial Comorbidities.
Differences across the HIV care continuum between men who have sex with men and women (MSMW) and men who have sex with men only (MSMO) are emerging in recent literature but have not been comprehensively documented among black MSM. Although MSMW have lower HIV prevalence than MSMO, they are more likely to be HIV-positive unaware and be virally unsuppressed. Explanatory factors for these differences have not previously been assessed. Between 2014 and 2016, we surveyed sexually active black MSM 18 years or older at Black Gay Pride events in 6 U.S. cities (n = 3881), 1229 of whom either self-reported HIV-positive status or tested HIV-positive onsite. We compared HIV-positive MSMW (n = 196) with HIV-positive MSMO (n = 1033) by HIV-positive unaware status, HIV care uptake, and viral load suppression. We conducted multivariable logistic regressions and built a structural equation model assessing mediating effects of psychosocial comorbidities (violence victimization, depression, and polydrug use) on the relationship between MSMW status and unsuppressed virus. MSMW were more likely than MSMO to be HIV-positive unaware [adjusted odds ratio (aOR) = 2.17; 95% confidence interval (CI): 1.58 to 3.00]. Among those who were HIV-positive aware (n = 720), MSMW were more likely to report never receiving HIV care (aOR = 2.74; 95% CI: 1.05 to 7.16) and to report detectable viral loads (aOR = 2.34; 95% CI: 1.31 to 4.19). Psychosocial comorbidities significantly mediated (P = 0.01) the relationship between MSMW status and unsuppressed virus. Black MSMW were less likely than black MSMO to uptake biomedical care and secondary prevention. Biobehavioral intervention development specific to HIV-positive black MSMW will be most successful if psychosocial comorbidities are also addressed.


A trans-disciplinary study on the health risks of cryptosporidiosis from dairy systems in Dagoretti, Nairobi, Kenya: study background and farming system characteristics.
This paper characterises the dairy farming system in Dagoretti, Nairobi. Characterisation was part of a broader ecohealth project to estimate the prevalence and risk of cryptosporidiosis and develop risk mitigation strategies. In the project a trans-disciplinary team addressed epidemiological, socioeconomic, environmental and policy aspects of cryptosporidiosis, an emerging zoonosis. This paper also provides background and describes sampling methods for the wider project. Three hundred dairy households were probabilistically sampled from a sampling frame of all dairy households in five of the six locations of Dagoretti, one of the eight districts of Nairobi Province. Randomly selected households identified 100 non-dairy-keeping households who also took part in the study. A household questionnaire was developed, pre-tested and administered in the dry and wet seasons of 2006. An additional study on livelihood and economic benefits of dairying took place with 100 dairy farmers randomly selected from the 300 farms (as well as 40 non-dairy neighbours as a control group), and a risk-targeted survey of environmental contamination with Cryptosporidium was conducted with 20 farmers randomly selected from the 29 farmers in the wider survey who were considered at high risk because of farming system. We found that around 1 in 80 urban households kept dairy cattle with an average of three cattle per household. Cross-breeds of exotic and local cattle predominate. Heads of dairy-keeping households were significantly less educated than the heads of non-dairy neighbours, had lived in Dagoretti for significantly longer and had significantly larger households. There was a high turnover of 10 % of the cattle population in the 3-month period of the study. Cattle were zero grazed, but productivity parameters were sub-optimal as were hygiene and husbandry practices. In conclusion, dairy keeping is a minor activity in urban Nairobi but important to households involved and their community. Ecohealth approaches are well suited to tackling the complex problem of assessing and managing emerging zoonoses in urban settings.


NOD-like receptors mediated activation of eosinophils interacting with bronchial epithelial cells: a link between innate immunity and allergic asthma.
Key intracytosolic pattern recognition receptors of innate immunity against bacterial infections are nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs). We elucidated the NOD1 and NOD2-mediated activation of human eosinophils, the principal effector cells for allergic inflammation, upon interacting with human bronchial epithelial BEAS-2B cells in allergic asthma. Eosinophils constitutively expressed NOD1,2 but exhibited nonsignificant responses to release chemokines upon the stimulation by NOD1 ligand γ-D-glutamyl-meso-diaminopimelic acid (iE-DAP) and NOD2 ligand muramyl dipeptide (MDP). However, iE-DAP and MDP could significantly upregulate cell surface expression of CD18 and intercellular adhesion molecule (ICAM)-1 on eosinophils and ICAM-1 on BEAS-2B cells, as well as induce chemokines CCL2 and CXCL8 release in the coculture system (all P<0.05). Both eosinophils and BEAS-2B cells were the main source for CXCL8 and CCL2 release in the coculture system upon iE-DAP or MDP stimulation. Direct interaction between eosinophils and BEAS-2B cells is responsible for CCL2 release, and soluble mediators are implicated in CXCL8 release. ERK and NF-κB play regulatory roles for the expression of adhesion molecules and chemokines in coculture. Treatment with NOD1,2 ligand could induce the subepithelial fibrosis and significantly enhance the serum concentration of total IgE, chemokine CCL5 for eosinophils and T helper type 2 (Th2) cells and asthma Th2 cytokine IL-13 in bronchoalveolar lavage fluid of ovalbumin-sensitized allergic asthmatic mice (all P<0.05). This study provides further evidence of bacterial infection-mediated activation of NOD1,2 in triggering allergic asthma via the activation of eosinophils interacting with bronchial epithelial cells at inflammatory airway.


Modulation of respiratory dendritic cells during Klebsiella pneumonia infection.
Klebsiella pneumoniae is a leading cause of severe hospital-acquired respiratory tract infections and death but little is known regarding the modulation of respiratory dendritic cell (DC) subsets. Plasmacytoid DC (pDC) are specialized type 1 interferon producing cells and considered to be classical mediators of antiviral immunity. By using multiparameter flow cytometry analysis we have analysed the modulation of respiratory DC subsets after intratracheal Klebsiella pneumonia infection. Data indicate that pDCs and MoDC were markedly elevated in the post acute pneumonia phase when compared to mock-infected controls. Analysis of draining mediastinal lymph nodes revealed a rapid increase of activated CD103+ DC, CD11b+ DC and MoDC within 48 h post infection. Lung pDC identification during bacterial pneumonia was confirmed by extended phenotyping for 120G8, mPDCA-1 and Siglec-H expression and by demonstration of high Interferon-alpha producing capacity after cell sorting. Cytokine expression analysis of ex vivo-sorted respiratory DC subpopulations from infected animals revealed elevated Interferon-alpha in pDC, elevated IFN-gamma, IL-4 and IL-13 in CD103+ DC and IL-19 and IL-12p35 in CD11b+ DC subsets in comparison to CD11c+ MHC-class IIlow cells indicating distinct functional roles. Antigen-specific naive CD4+ T cell stimulatory capacity of purified respiratory DC subsets was analysed in a model system with purified ovalbumin T cell receptor transgenic naive CD4+ responder T cells and respiratory DC subsets, pulsed with ovalbumin and matured with Klebsiella pneumoniae lysate. CD103+ DC and CD11b+ DC subsets represented the most potent naive CD4+ T helper cell activators. These results provide novel insight into the activation of respiratory DC subsets during Klebsiella pneumonia infection. The detection of increased respiratory pDC numbers in bacterial pneumonia may indicate possible novel pDC functions with respect to lung repair and regeneration.


Bupivacaine wound infiltration in thyroid surgery reduces postoperative pain and opioid demand.
Control of postoperative pain is an important element in preventing the modification of the excitability of the dorsal horn neurons. We studied the efficacy of bupivacaine 0.5% wound infiltration for postoperative pain management following thyroid surgery. Forty consecutive ASA I-II patients, scheduled for thyroidectomy, were assigned randomly to two groups. Group I (n = 20) consisted of patients receiving bupivacaine 0.5% (10 ml) wound infiltration at the end of surgery and group II (n = 20 included patients without infiltration. The patients did not know whether the wound had been anaesthetized or not. All patients received balanced inhalational anaesthesia, including fentanyl (a total dose of up to 4 micrograms.kg-1). Postoperative pain medication included morphine IV or IM, as needed. Twenty-four hours after surgery the worst pain was recorded using a visual analogue scoring system, from 0 to 10. Twenty-four hour postoperative morphine requirement was recorded. Groups did not differ in demographic data. Pain scores significantly different in the two groups. In group I, the mean pain score was 3.7 +/- 1.6 compared with 6.9 +/- 1.7 in group II (P < 0.05). Only six patients (30%) in group I received opioids and only one of these (5%) had a pain score above 5. In comparison, 18 patients (90%) in group II received morphine during the first postoperative day. The local injection of bupivacaine corresponds to a block of the superficial branches of the cervical plexus. This study demonstrated a simple, efficient and safe way to reduce pain perception following a thyroidectomy.


Immune response with biodegradable nanospheres and alum: studies in rabbits using staphylococcal enterotoxin B-toxoid.
In this study, the adjuvant effect of the sustained release biodegradable nanospheres (100-150 nm in diameter) has been compared with alum. Nanospheres were formulated using a biodegradable polylactic polyglycolic acid copolymer (PLGA, 50:50) containing Staphylococcal Enterotoxin B (SEB) toxoid as a model vaccine antigen. Systemic immune response of the nanospheres containing toxoid was studied in rabbits by subcutaneous immunization. The data demonstrated that approximately 30% of the toxoid activity was lost following its encapsulation into nanospheres. Under in vitro conditions, nanospheres demonstrated sustained release of the toxoid. However, only 20% of the antigenic toxoid was released over the first 2 weeks of the release study. Immunization of animals with equal doses of toxoid, either using nanospheres or alum induced a comparable systemic immune response (IgG, IgM and IgA). The immune response reached a maximum level at 7 weeks post-immunization, which then gradually declined with time. The booster dose of toxoid at 19 weeks, either using alum or nanospheres induced similar immune response in both the groups, but was greater than the primary immune response. The studies, thus, suggest that biodegradable nanospheres could be used as a vaccine adjuvant.


Induction of the nitric oxide-synthesizing pathway in fresh and interleukin 2-cultured rat natural killer cells.
Several lines of evidence suggest that nitric oxide (NO), generated through nitric oxide synthase (NOS) by cleavage of terminal guanidino nitrogen from L-arginine, mediates tumor cell killing by mononuclear phagocytes. Natural killer (NK) cells are cytotoxic effector cells that lyse a variety of tumor and virus-infected cells in a MHC-unrestricted manner. NK cells cultured with interleukin 2 proliferate and acquire the ability to lyse a wide range of targets, including NK-resistant tumor cells (LAK activity). The present study was designed to investigate whether a NOS pathway exists in fresh or IL-2-activated NK cells and to assess the importance of NO synthesis in their activation and cytotoxic functions. NKR-P1 triggering, which is known to induce NK cell activation and mediate reverse ADCC, was able to induce arginine metabolism with consequent increase of nitrite and citrulline levels. Moreover, stimulated NO synthesis leads to guanylate cyclase activity with consequent cGMP generation. We also report that cytotoxic activities of fresh or IL-2-activated NK cells appear to be dependent on arginine levels in medium. Tumoricidal activity of both these effector cells, assessed against YAC-1 and P815 target cells, respectively, was indeed significantly reduced when cytotoxic assays were performed in arginine-free medium or in the presence of the L-arginine analog L-N-monomethyl-arginine, which inhibits nitroxide formation from L-arginine. Normal levels of cytotoxic activities could be restored by addition of exogenous L-arginine. NO generation by NK and LAK cells, determined as nitrite, citrulline, and cGMP synthesis, correlated well with their cytotoxic activities. Moreover, NOS activity gradually increased during the LAK generation and correlated well with the increasing capability of IL-2-activated NK cells to lyse NK-resistant targets, such as P815.


Chemotactic properties, cellular binding and uptake of peptides and peptide derivatives: studies with Tetrahymena thermophila.
Receptor-mediated binding of leukocyte chemotactic peptide, N-formylMet-Leu-Phe (fMLP), occurs in the ciliated protozoon Tetrahymena thermophila. In vivo labelling of the cells with N-formylMet-Leu-[3H]Phe ([3H]fMLP) shows that the cells bind the ligand with high affinity (KD = 4 x 10(-9) M to 1 x 10(-8) M). Moreover, Scatchard transformations of the binding data show that there are about 5 x 10(5) binding sites per cell on the cell surface. Two fluorescent derivatives of leukocyte chemotactic peptide, N-dansylMet-Leu-Phe (dansMLP) and N-formylMet-Leu-Phe-(N-dansyl-)Lys (fMLPdanLys) compete for the N-formylMet-Leu-Phe (fMLP) binding sites on the cell surface. Moreover, both derivatives have retained significant chemoattracting potentials. Fluorescence from dansMLP, but not from fMLPdansLys and dansyl-beta-endorphin, is internalized preferentially into small vesicles. The differences may, however, reflect that the fluorescence from the dansyl group is strongly quenched by a hydrophilic microenvironment when using the two latter peptide derivatives. In contrast, the dansyl group from dansMLP must be assumed to be embedded in a hydrophobic microenvironment in the vesicular membrane or membrane protein. Rhodamine-labelled bovine serum albumin, egg albumin and cytochrome c as well as dansylated bovine serum albumin, which are poor chemoattractants, are preferentially seen to be internalized into large vesicles (food vacuoles).


Cooperative action of glutamate transporters and cystine/glutamate antiporter system Xc- protects from oxidative glutamate toxicity.
Oxidative glutamate toxicity in the neuronal cell line HT22 is a model for cell death by oxidative stress. In this paradigm, an excess of extracellular glutamate blocks the glutamate/cystine-antiporter system Xc-, depleting the cell of cysteine, a building block of the antioxidant glutathione. Loss of glutathione leads to the accumulation of reactive oxygen species and eventually cell death. We selected cells resistant to oxidative stress, which exhibit reduced glutamate-induced glutathione depletion mediated by an increase in the antiporter subunit xCT and system Xc- activity. Cystine uptake was less sensitive to inhibition by glutamate and we hypothesized that glutamate import via excitatory amino acid transporters and immediate re-export via system Xc- underlies this phenomenon. Inhibition of glutamate transporters by l-trans-pyrrolidine-2,4-dicarboxylic acid (PDC) and DL-threo-beta-benzyloxyaspartic acid (TBOA) exacerbated glutamate-induced cell death. PDC decreased intracellular glutamate accumulation and exacerbated glutathione depletion in the presence of glutamate. Transient overexpression of xCT and the glutamate transporter EAAT3 cooperatively protected against glutamate. We conclude that EAATs support system Xc- to prevent glutathione depletion caused by high extracellular glutamate. This knowledge could be of use for the development of novel therapeutics aimed at diseases associated with depletion of glutathione like Parkinson's disease.


Comparison of digitized images with original radiography for semiquantitative assessment of osteoporotic fractures.
We validated a vertebral fracture assessment (VFA) workstation developed by our group for semiquantitative assessment of vertebral fractures in large-scale, multicenter osteoporosis drug trials. Baseline and follow-up spine radiographs (lateral views) of 50 patients who participated in a clinical trial were digitized and were archived on CD-ROM. Both original radiographs and the digitized images were independently assessed by three experienced radiologists. Prevalent fracture scores of vertebrae were rated in increments of 1 on a 4-point scale. Incident fractures were defined as any worsening of grade on follow-up films. Generally good to excellent agreement among the three readers was found between the two methods, with kappa scores (kappa) from 0.91 to 0.96 for prevalence of fractures, and from 0.80 to 0.90 for incidence of fractures. Reproducibility (intra-reader variability) of each method was comparable. For assessing prevalent fracture, kappa was from 0.87 to 0.96 using radiographs, and from 0.87 to 0.94 using VFA images. For incident fractures, the kappa was from 0.78 to 0.89 using radiographs, and from 0.82 to 0.88 using VFA images. Level-specific agreement between the two approaches was consistent. Overall, there is no difference between readings of digital images and readings of conventional radiographs. The quality of the new VFA for visualization of vertebral fracture is excellent.


Cardioprotective effects of fucoidan against hypoxia-induced apoptosis in H9c2 cardiomyoblast cells.
Cardiomyocyte apoptosis plays a critical role in the progress of heart diseases. Fucoidan, a complex-sulfated polysaccharide, has been reported to possess potential cardioprotective efficacy in vivo. The present study determines whether fucoidan could provide cardioprotection on hypoxia-induced cardiomyocyte apoptosis. H9c2 cardiomyoblast cells were incubated with various concentrations (15, 30, and 60 μg/ml) of fucoidan in a humidified incubator at 37 °C with 95% O2 and 5% CO2. After 6 h, hypoxia was processed and the cardioprotective effects of fucoidan were evaluated by applying MTT, ELISA, Hoechst 33258 nucleus staining, and western blot. Following a 6 h exposure of H9c2 to hypoxic condition, significant reduction was found in cell survival (0.57-fold) and superoxide dismutase (SOD) activity (0.56-fold), which were associated with the increase of malondialdehyde (MDA) level (2.58-fold), creatine phosphokinase (CK, 3.57-fold), and lactate dehydrogenase (LDH) activities (2.39-fold). Moreover, hypoxia-induced apoptosis was confirmed by Hoechst 33258 nuclear staining, and these changes were accompanied by the increase of Bcl-2 (1.27-fold) and Bax expression (2.6-fold). However, preincubation of the cells with fucoidan prior to hypoxia exposure elevated the cell viability (30 μg/ml, 1.18-fold; 60 μg/ml, 1.32-fold) and SOD activity (30 μg/ml, 1.12-fold; 60 μg/ml, 1.25-fold), but decreased the MDA level (30 μg/ml, 0.70-fold; 60 μg/ml, 0.80-fold), CK (30 μg/ml, 0.69-fold; 60 μg/ml, 0.76-fold), and LDH (30 μg/ml, 0.67-fold; 60 μg/ml, 0.86-fold) leakages. Hoechst 33258 nuclear staining observations demonstrated the same protective effect of fucoidan on hypoxia-induced myocardial injury. Also, cardioprotective effects of fucoidan were reflected by increasing Bcl-2 (60 μg/ml, 1.84-fold), as well as decreasing Bax (60 μg/ml, 0.6-fold). Fucoidan had protective effect against hypoxia-induced cardiomyocytes apoptosis, and the mechanism might involve protections of the cell from oxidative injury.


Endotoxin stimulates in vitro pituitary growth hormone release in eicosanoid-dependent manner.
To investigate the signal transduction pathways by which endotoxin stimulates in vitro pituitary cell growth hormone (GH) release. Pituitary cell cultures derived from 6 sheep. Signal transduction pathways involved in endotoxin-mediated GH release from sheep pituitary cell cultures were evaluated by the use of specific blockers of arachidonic acid and its metabolites, extracellular calcium, protein kinase C, and protein kinase A. Cell cultures were exposed to the specific blockers in the presence or absence of endotoxin (Escherichia coli O55:B5, 10 micrograms/ml) for 24 hours. In addition, effects of endotoxin on GH cell content and GH mRNA values were determined. Nordihydroquairetic acid (lipoxygenase blocker, 10 microM, 30 microM) and eicosatetraynoic acid (arachidonic acid competitor, 10 microM) decreased endotoxin-stimulated GH release. The calcium channel blocker verapamil (25 microM) decreased baseline and endotoxin-stimulated GH release. Phorbol myristate acetate-induced down-regulation of protein kinase C, indomethacin, or the protein kinase A blocker H89 did not alter endotoxin-stimulated GH release. Endotoxin increased GH mRNA values by 50.1 +/- 6.0%, but the cell content of GH was not affected. A direct effect of endotoxin on the pituitary gland to stimulate GH secretion was evident, an effect mediated predominantly by arachidonic acid and its metabolites through the lipoxygenase pathway. Endotoxin-stimulated GH release requires extracellular calcium and is associated with increased cell GH mRNA content. A better understanding of the signal transduction pathways involved in endotoxin-mediated effects will allow more appropriate therapeutic intervention in clinical cases of endotoxemia.


Extracellular GABA concentrations in rat supraoptic nucleus during lactation and following haemodynamic changes: an in vivo microdialysis study.
Morphological and pharmacological evidence suggest that the dense GABAergic innervation of the supraoptic nucleus is important for regulating the electrical activity of vasopressin and oxytocin neurons. We have employed the technique of intracranial microdialysis to examine extracellular GABA concentrations in the supraoptic nucleus of the anaesthetized rat and questioned whether differences exist in the dynamics of GABA release between virgin and lactating rats, and if events during lactation or following blood pressure manipulation alter endogenous GABA levels in this nucleus. No significant differences were detected between virgin and lactating animals in either basal or 100 mM potassium ion-evoked GABA release. The inclusion of the GABA uptake blocker nipecotic acid (0.5 mM) into the dialysate resulted in a six- to eight-fold increase (P < 0.01) in GABA outflow in both groups of animals. In lactating rats, GABA outflow measured at 4 min intervals was not altered during a 60 min period of suckling by a full litter of pups and no significant change in GABA outflow was detected in relation to individual milk ejections. In virgin rats, removal of 1.5-2 ml of blood resulted in a 30-60 mmHg fall in blood pressure and a non-significant decline in GABA outflow. Replacement of blood resulted in an abrupt 50 mmHg increase in blood pressure and a significant 22% increase in GABA outflow (P < 0.01), but no change in aspartate or methionine concentrations. Repeated intravenous injections of the alpha-adrenoceptor agonist, metaraminol, similarly evoked approximately 50 mmHg increments in blood pressure and a 26% increase in GABA outflow (P < 0.05). Electrical stimulation of the diagonal band of Broca for 10 min produced a two-fold increase in GABA outflow from the supraoptic nucleus (P < 0.05). These results show that the overall profile of basal and potassium-stimulated GABA concentrations in the supraoptic nucleus is not substantially different between lactating and virgin rats. In lactating animals we have found that GABA levels are not altered in response to suckling or at the time of high-frequency firing by oxytocin neurons to induce milk ejection. In contrast, our data further support the hypothesis that GABA inputs to supraoptic neurons are part of a baroreceptor reflex, relaying through the diagonal band of Broca, to signal periods of acute hypertension and inhibit the firing of vasopressin neurons. Such observations suggest the physiological importance of GABA inputs to the supraoptic nuclei and indicate that GABA may be used in a stimulus-specific manner to influence the activity of magnocellular neurons.


Diverse roles of RNA polymerase II-associated factor 1 complex in different subpathways of nucleotide excision repair.
Transcription-coupled repair (TCR) and global genomic repair (GGR) are two pathways of nucleotide excision repair (NER). In Saccharomyces cerevisiae, Rad26 is important but not absolutely required for TCR. Rpb4, a nonessential RNA polymerase II (Pol II) subunit that forms a subcomplex with Rpb7, and the Spt4-Spt5 complex, a transcription elongation factor, have been shown to suppress Rad26-independent TCR. The Pol II-associated factor 1 complex (Paf1C) has been shown to function in transcription elongation, 3'-processing of mRNAs, and posttranslational modification of histones. Here we show that Paf1C plays a marginal role in facilitating Rad26-dependent TCR but significantly suppresses Rad26-independent TCR. The suppression of Rad26-independent TCR is achieved by cooperating with Spt4-Spt5. We propose a model that, in the absence of Rad26, a lesion is "locked" in the active center of a Pol II elongation complex, which is stabilized by the coordinated interactions of Rpb4-Rpb7, Spt4-Spt5, and Paf1C with each other and with the core Pol II. We also found that Paf1C facilitates GGR, especially in internucleosomal linker regions. The facilitation of GGR is achieved through enabling monoubiquitination of histone H2B lysine 123 by Bre1, which in turn permits di- and trimethylation of histone H3 lysine 79 by Dot1. To our best knowledge, among the NER-modulating factors documented so far, Paf1C appears to have the most diverse functions in different NER pathways or subpathways.


Phosphorylation of cardiac regulatory proteins by cyclic AMP-dependent protein kinase.
Cardiac myofibrils were purified from canine myocardium, and the regulatory proteins (troponin + tropomyosin) were extracted and shown to contain endogenous cyclic AMP-dependent protein kinase activity. Other cyclic nucleotide stimulated the protein kinase activity but only at higher concentrations. The enzyme was able to catalyze phosphorylation of conventional substrates such as histones and casein as well as a component of the regulatory protein fraction with a molecular weight of 28,000 daltons. Endogenous phosphorylation required the presence of Mg2+ and was inhibited by Ca2+. A protein kinase inhibitor obtained from skeletal muscle inhibited the cyclicAMP-dependent phosphorylation. Escherichia coli alkaline phosphatase dephosphorylated the endogenous substrates. The level of phosphorylation found is severalfold higher than we have previously reported. A protein kinase, with its close association with the regulatory proteins, seems to be well suited to transmitting the message from the cyclic AMP to the regulatory proteins, a phenomenon that may influence the cardiac contractility via the troponin phosphorylation. The inhibitory effect of troponin on actomyosin might be changed by its state of phosphorylation.


Incidental finding of sacral hemiagenesis on the (99m)Tc-MDP bone scan as a casual regression syndrome type II.
A 69 years old woman with chronic low back pain referred to our Department for bone scintigraphy. Patient did not have any other complaint and his physical examination of the patient was normal. Whole-body scan was acquired 3 h after the intravenous injection of 740 MBq of technetium-99m methylene diphosphonate ((99m)Tc-MDP). Distal sacrococcygeal region could not be observed during the visual analysis of the whole-body scan. Pelvic X-rays radiography and pelvic computed tomography of the patient, demonstrated hemiagenesis of the sacrum, which was consistent with type-2 sacral agenesis. Other structural abnormalities were not detected on the pelvic CT scan of the patient. This case is presented to demonstrate the rare congenital anomaly of sacral hemiagenesis causing empty pelvis appearance in the posterior projection of (99m)Tc-MDP whole body bone scan. This congenital anomaly could be associated with extensive abnormalities of the lower vertebrae, pelvis, and spine.


A novel and potentially valuable exposure measure: Escherichia coli in oral cavity and its association with child daycare center attendance.
This study investigated the occurrence of Escherichia coli, an indicator of fecal contamination, in saliva samples from 141 children up to 36 months old, 53 not attending and 88 attending a child daycare center after exposure to a public daycare center and home environments. Two samples from each child were collected on the same day at 07:00 and 15:00 h and plated on MacConkey agar for identification. Samples E. coli negative in the morning and positive in the afternoon were statistically associated with the condition of the child attending daycare center (Odds ratio = 2.72; 95% confidence interval = 1.15/6.46). Exposure to the daycare center environment favored the potential risk of transmission of enteropathogens, as demonstrated by the presence of E. coli in saliva. The method proved to be easy to sample, non-invasive and feasible in young children. The findings suggest a novel and potentially valuable exposure measure.


Structural features of ring C of 20-oxo steroids and the interaction with cortisone reductase.
Kinetic measurements were made with cortisone reductase (20-dihydrocortisone-NAD(+) oxidoreductase, EC 18.104.22.168) and a series of substrates which differed in shape, size and electronic character in the region adjacent to C-11, C-14 and C-18. Structural changes at C-11 in these substrates resulted in up to 660-fold changes in the apparent K(m) value, up to 200-fold changes in the apparent V(max.) value and up to 800-fold changes in the ratio of these kinetic constants. It is suggested that interactions important for substrate function normally occur between the enzyme and the C ring in the region of C-11, that these interactions arise from so-called hydrophobic forces between the generally hydrophobic C ring portion of the substrate and a hydrophobic region of the enzyme, but that when the substrate contains a polar substituent in this portion of the molecule, then polar interactions with polar moieties of the enzyme can also be important. It is further suggested that the part of the enzyme that interacts with the region of C-11 in the substrate is flexible, and that substrate binding involves at least some degree of induced fit.


[Squamous cell carcinoma of the rectum and traumatic recto vaginal fistula relation or coincidence? Case study].
Primary squamous cell carcinoma of the rectum is rare and its cause and pathogenesis are not clear. Likewise, rectal squamous cell carcinoma in patients with rectovaginal fistula in the absence of gynecologic cancer is extremely rare. We report one of such cases that was diagnosed a year after an untreated traumatic rectovaginal fistula. We also reviewed the literature and discussed the probable association between both processes.


Platelet-derived growth factor receptors in the kidney--upregulated expression in inflammation.
Major features of a long-standing inflammation in the kidney are vascular proliferation, glomerulosclerosis, interstitial fibrosis and tubular atrophy, leading to a gradual deterioration of the renal function. In this study we have investigated the expression of B-type receptors for platelet-derived growth factor (PDGF) in frozen sections from normal and inflamed kidneys. Immunohistochemical techniques, employing two monoclonal antibodies specific for PDGF B-type receptors, were used. The specimens investigated were 15 kidneys removed by transplantectomy because of chronic rejection, 20 cases of glomerulonephritis with crescent formation, mesangial proliferation or non-proliferative glomerulonephritis, and six normal kidneys. In parallel we characterized cellular infiltrates and class II transplantation antigen expression in the inflamed kidneys. An enhanced PDGF receptor expression was found on intimal cells and on smooth muscle cells of the proliferating vessels, on glomerular cells in glomeruli with mesangial proliferation, and on fibroblast-like cells in the proximity of clusters of infiltrating macrophages and T-lymphocytes of the interstitial tissue. Induction of PDGF receptor expression may render cells responsive to stimulation by PDGF, released from PDGF-producing cells, such as activated macrophages and from platelets. Our data suggest that PDGF is involved in the proliferation of mesenchymal cells that is seen in rejected kidney transplants and glomerulonephritis.


A role for nonsense-mediated mRNA decay in plants: pathogen responses are induced in Arabidopsis thaliana NMD mutants.
Nonsense-mediated mRNA decay (NMD) is a conserved mechanism that targets aberrant mRNAs for destruction. NMD has also been found to regulate the expression of large numbers of genes in diverse organisms, although the biological role for this is unclear and few evolutionarily conserved targets have been identified. Expression analyses of three Arabidopsis thaliana lines deficient in NMD reveal that the vast majority of NMD-targeted transcripts are associated with response to pathogens. Congruently, NMD mutants, in which these transcripts are elevated, confer partial resistance to Pseudomonas syringae. These findings suggest a biological rationale for the regulation of gene expression by NMD in plants and suggest that manipulation of NMD could offer a new approach for crop protection. Amongst the few non-pathogen responsive NMD-targeted genes, one potential NMD targeted signal, the evolutionarily conserved upstream open reading frame (CuORF), was found to be hugely over-represented, raising the possibility that this feature could be used to target specific physiological mRNAs for control by NMD.


Diagnostic yield of specific inhalation challenge in hypersensitivity pneumonitis.
Reliable methods are needed to diagnose hypersensitivity pneumonitis. The aim of the study was to establish the diagnostic yield of specific inhalation challenge (SIC) in patients with hypersensitivity pneumonitis. All patients with suspected hypersensitivity pneumonitis in whom SIC was performed (n=113) were included. SIC was considered positive when patients showed a decrease of >15% in forced vital capacity (FVC) or >20% in diffusing capacity of the lung for carbon dioxide, or a decrease of 10% to 15% in FVC accompanied by a temperature increase of 0.5°C within 24 h of inhalation of the antigen. SIC was positive to the agents tested in 68 patients: 64 received a diagnosis of hypersensitivity pneumonitis and SIC results were considered false-positive in the remaining four patients. In the SIC-negative group (n=45), 24 patients received a diagnosis of hypersensitivity pneumonitis and SIC results were considered false-negative, and 21 patients were diagnosed with other respiratory diseases. The sensitivity and specificity of the test were 72.7% and 84%, respectively. Having hypersensitivity pneumonitis caused by an antigen other than birds or fungi predicted a false-negative result (p=0.001). In hypersensitivity pneumonitis, positive SIC testing virtually confirms the diagnosis, whereas negative testing does not rule it out, especially when the antigenic sources are not birds or fungi.


Probing the Cys-Tyr Cofactor Biogenesis in Cysteine Dioxygenase by the Genetic Incorporation of Fluorotyrosine.
Cysteine dioxygenase (CDO) is a nonheme iron enzyme that adds two oxygen atoms from dioxygen to the sulfur atom of l-cysteine. Adjacent to the iron site of mammalian CDO, there is a post-translationally generated Cys-Tyr cofactor, whose presence substantially enhances the oxygenase activity. The formation of the Cys-Tyr cofactor in CDO is an autocatalytic process, and it is challenging to study by traditional techniques because the cross-linking reaction is a side, uncoupled, single-turnover oxidation buried among multiple turnovers of l-cysteine oxygenation. Here, we take advantage of our recent success in obtaining a purely uncross-linked human CDO due to site-specific incorporation of 3,5-difluoro-l-tyrosine (F2-Tyr) at the cross-linking site through the genetic code expansion strategy. Using EPR spectroscopy, we show that nitric oxide (•NO), an oxygen surrogate, similarly binds to uncross-linked F2-Tyr157 CDO as in wild-type human CDO. We determined X-ray crystal structures of uncross-linked F2-Tyr157 CDO and mature wild-type CDO in complex with both l-cysteine and •NO. These structural data reveal that the active site cysteine (Cys93 in the human enzyme), rather than the generally expected tyrosine (i.e., Tyr157), is well-aligned to be oxidized should the normal oxidation reaction uncouple. This structure-based understanding is further supported by a computational study with models built on the uncross-linked ternary complex structure. Together, these results strongly suggest that the first target to oxidize during the iron-assisted Cys-Tyr cofactor biogenesis is Cys93. Based on these data, a plausible reaction mechanism implementing a cysteine radical involved in the cross-link formation is proposed.


Antibody-Based Fusion Proteins Allow Ca2+ Rewiring to Most Extracellular Ligands.
The Ca2+ signaling toolkit is the set of proteins used by living systems to generate and respond to Ca2+ signals. The selective expression of these proteins in particular tissues, cell types and subcellular locations allows the Ca2+ signal to regulate a diverse set of cellular processes. Through synthetic biology, the Ca2+ signaling toolkit can be expanded beyond the natural repertoire to potentially allow a non-natural ligand to control downstream cellular processes. To realize this potential, we exploited the ability of an antibody to bind its antigen exclusively in combination with the ability of the cytoplasmic domain of vascular endothelial growth factor receptor 2 (VEGFR2) to generate a Ca2+ signal upon oligomerization. Using protein fusions between antibody variants (i.e., nanobody, single-chain antibody and the monoclonal antibody) and the VEGFR2 cytoplasmic domain, Ca2+ signals were generated in response to extracellular stimulation with green fluorescent protein, mCherry, tumor necrosis factor alpha and soluble CD14. The Ca2+ signal generation by the stimulus did not require a stringent transition from monomer to oligomer state, but instead only required an increase in the oligomeric state. The Ca2+ signal generated by these classes of antibody-based fusion proteins can be rewired with a Ca2+ indicator or with an engineered Ca2+ activated RhoA to allow for antigen screening or migration to most extracellular ligands, respectively.


The murine (C3H/He) epidermal Ia+ dendritic cells (Ia+DECs) and Thy-1+ dendritic cells (Thy-1+DECs) in contact hypersensitivity and aging.
The contact sensitivity evaluated by the ear swelling test and the dynamic changes of epidermal Ia+ dendritic cells (Ia+DECs) and Thy-1+ dendritic cells (Thy-1+DECs) were studied in trinitrochlorobenzene (TNCB) sensitized different age group C3H/He mice after challenge. A significant increase of ear swelling was observed between 6 h and 10 days of both 8-10 week (wk) and 40-48 wk groups; the ear swelling indices of 8-10 wk group were significantly higher than those of 40-48 wk group from 18 h to 5 days. A significant decrease of the densities of Ia+DECs from 18 h to 48 h, followed by a gradual increase reaching significant increase of the densities of Ia+DECs from 5 days to 21 days in both 8-10 wk and 40-48 wk groups, was observed; the densities of Thy-1+DECs significantly decreased from 18-48 h, followed by a gradual increase reaching a significant increase from 5 days to 21 days in both 8-10 wk and 40-48 wk groups. In the normal control groups, a significant decline of both Ia+DECs and Thy-1+DECs in the 40-48 wk group was observed. Results suggest that contact allergy may be diminished in aged mice. On the other hand, like Ia+DECs, Thy-1+DECs seem to be involved in the process of contact allergy.


[What may be learnt from short- and long term tests and from studies on cancerogenicity, teratogenicity and mutagenicity (author's transl)].
During the last 15 years the transferability of results from animal experiments to humans has been improved so that today certainly more than 70% of possible side effects can be predicted. Systemic hypersensitivity reactions cannot be predicted, however, since to date there are no safe animal experimental methods available. In spite of many decades of research in the field of animal experimental cancerogenesis the value of investigations of this kind for humans can be limited by the selection of unsuitable animal species, use of inadequate modes of administration or excessive doses. Similar aspects apply to reproduction toxicology; there are many examples in cancerology and teratology for false-positive and -negative results from animal experiments. In chemomutagenesis comparisons of in vivo cytogenetic investigations in somatic cells of animals and humans have revealed largely concurring results. However, only stable mutations found in generative cells are signs of a genetic danger. It will therefore have to be clarified which conclusions for generative cells should be drawn from changes in somatic cells. Furthermore, the elaboration of methods for the determination of the mutation of genes in the organism of mammals still requires intensive research.


Characterization of silver nanoparticles synthesized on titanium dioxide fine particles.
Silver nanoparticles with a narrow size distribution were synthesized over the surface of two different commercial TiO(2) particles using a simple aqueous reduction method. The reducing agent used was NaBH(4); different molar ratios TiO(2):Ag were also used. The nanocomposites thus prepared were characterized using transmission electron microscopy (TEM), scanning transmission electron microscopy (STEM), scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDS), x-ray photoelectron spectroscopy (XPS), x-ray diffraction (XRD), dynamic light scattering (DLS) and UV-visible (UV-vis) absorption spectroscopy; the antibacterial activity was assessed using the standard microdilution method, determining the minimum inhibitory concentration (MIC) according to the National Committee for Clinical Laboratory Standards. From the microscopy studies (TEM and STEM) we observed that the silver nanoparticles are homogeneously distributed over the surface of TiO(2) particles and that the TiO(2):Ag molar ratio plays an important role. We used three different TiO(2)Ag molar ratios and the size of the silver nanoparticles is 10, 20 and 80 nm, respectively. It was found that the antibacterial activity of the nanocomposites increases considerably comparing with separated silver nanoparticles and TiO(2) particles.


Large-Scale Proteomic Identification of Targets of Cellular miR-197 Downregulated by Enterovirus A71.
MicroRNAs are noncoding RNA species comprising 18-23 nucleotides that regulate host-virus interaction networks. Here, we show that enterovirus A71 infection in human rhabdomyosarcoma (RD) is regulated by miR-197 expression. Transfection of miR-197 mimic into RD cells inhibited virus replication by interfering with the viral RNA synthesis. We employed a combination of mass-spectrometry-based quantitative proteomics with the stable isotope labeling with amino acids in cell culture (SILAC) approach for the identification of the miR-197 target genes in RD cells and to investigate the differential expression of the prospective target proteins. A total of 1822 proteins were repeatedly identified in miR-197-transfected RD cells, 106 of which were predicted to have seed sites by TargetScan. Notably, seven of eight selected genes potentially related to viral replication and immune response were validated as direct miR-197 targets, using a luciferase 3'-untranslated region (UTR) reporter assay. The expression levels of three selected endogenous molecules (ITGAV, ETF1, and MAP2K1/MEK1) were significantly reduced when RD cells were transfected with a miR-197 mimic. Our results provide a comprehensive database of miR-197 targets, which might provide better insights into the understanding of host-virus interaction.


International consensus classification of hippocampal sclerosis and etiologic diversity in children with temporal lobectomy.
The distribution of hippocampal sclerosis (HS) subtypes, according to the classification of the International League Against Epilepsy (ILAE), has been reported mainly in adult patients. We aimed to review the pathological findings in children who had anterior temporal lobectomy accompanied with amygdalohippocampectomy, in view of the current classification, and evaluate postsurgical outcome with respect to HS subtypes in childhood. Seventy children who underwent temporal resections for treatment of medically refractory epilepsy, with a minimum follow-up of 2 years, were included; the surgical hippocampus specimens were re-evaluated under the HS ILAE classification. Neuropathological evaluations revealed HS type 1 in 38 patients (54.3%), HS type 2 in 2 (2.8%), HS type 3 in 21 patients (30%), and no HS in 9 patients (12.9%). Of 70 patients, 23 (32.9%) had dual pathology, and the most common pattern was HS type 3 with low-grade epilepsy-associated brain tumors (LEAT). The distribution of HS types with respect to age revealed that HS type 3 and no HS subgroups had significantly more patients younger than 12 years, compared with those of HS type 1 (90.5%, 77.8% vs 47.4%, respectively). History of febrile seizures was higher in HS type 1. Prolonged/recurrent febrile seizures were most common in patients 12 years and older, whereas LEAT was the most common etiology in patients under 12 years of age (p < 0.001). Patients with HS type 1 had longer duration of epilepsy and an older age at the time of surgery compared with patients with HS type 3 and no HS (p: 0.031, p: 0.007). At final visit, 74.3% of the patients were seizure-free. Seizure outcome showed no significant difference between pathological subtypes. Our study presents the distribution of HS ILAE subtypes in an exclusively pediatric series along with long-term seizure outcome. The study reveals that the leading pathological HS subgroup in children is HS type 1, similar with adult series. Hippocampal sclerosis type 2 is significantly less in children compared with adults; however, HS type 3 emerges as the second most predominant group because of dual pathology, particularly LEAT. Further studies are required regarding clinicopathological features of isolated HS in pediatric cohort. Seizure-free outcome was favorable and similar in all HS types in children. The proportion of HS types may be better defined in pediatric patients with temporal resections, as the current HS ILAE classification becomes more widely used, and may help reveal the surgical and cognitive outcome with respect to HS types.


Controlling structure and porosity in catalytic nanoparticle superlattices with DNA.
Herein, we describe a strategy for converting catalytically inactive, highly crystalline nanoparticle superlattices embedded in silica into catalytically active, porous structures through superlattice assembly and calcination. First, a body-centered cubic (bcc) superlattice is synthesized through the assembly of two sets of 5 nm gold nanoparticles chemically modified with DNA bearing complementary sticky end sequences. These superlattices are embedded in silica and calcined at 350 °C to provide access to the catalytic nanoparticle surface sites. The calcined superlattice maintains its bcc ordering and has a surface area of 210 m(2)/g. The loading of catalytically active nanoparticles within the superlattice was determined by inductively coupled plasma mass spectrometry, which revealed that the calcined superlattice contained approximately 10% Au by weight. We subsequently investigate the ability of supported Au nanoparticle superlattices to catalyze alcohol oxidation. In addition to demonstrating that calcined superlattices are effective catalysts for alcohol oxidation, electron microscopy reveals preservation of the crystalline structure of the bcc superlattice following calcination and catalysis. Unlike many bulk nanoparticle catalysts, which are difficult to characterize and susceptible to aggregation, nanoparticle superlattices synthesized using DNA interactions offer an attractive bottom-up route to structurally defined heterogeneous catalysts, where one has the potential to independently control nanoparticle size, nanoparticle compositions, and interparticle spacings.


Estimating the incidence of enteric fever in children in India: a multi-site, active fever surveillance of pediatric cohorts.
Salmonella Typhi is responsible for about 20 million episodes of illness and over 140,000 deaths annually globally. South Asia has the highest documented burden of typhoid and is home to the multi-drug resistant H58 strain that makes treatment more challenging. The WHO recommends the use of Typhoid Conjugate Vaccines in typhoid endemic countries. Decisions on the preferred immunization strategy should be based on an analysis of disease burden, availability, affordability, and operational feasibility. Typhoid vaccines have so far remained unimplemented as public health measures because of a perceived decline in typhoid burden in recent years. The apparent decline, based on hospital reports, may be a result of rampant antimicrobial use in the community and therefore estimation of disease incidence at the community is necessary to better measure disease incidence and transmission. Age-specific incidence of typhoid fever in children between 6 months and 15 years will be estimated in four community based cohorts in varied settings across India using standardized protocols for active fever surveillance in the community. Data will be collected on secured cloud infrastructure using a combination of android and web-based real-time data collection tools. Blood cultures will be done for children with fever lasting 3 or more consecutive days using automated blood culture systems. Those with blood-culture confirmed typhoid fever will be followed up till 90 days to estimate costs and clinical outcomes of the illness episodes. Environmental factors, access to safe water, sanitation, hygiene, food hygiene, demography, population density and socioeconomic status will be assessed periodically to characterise risk factors and permit extrapolation of burden to similar risk settings. With the availability of licensed typhoid conjugated vaccines in India, it is important to consider whether the burden of disease is present and sufficient to require the use of vaccine in addition to other interventions. Active case finding in the community permits the detection of cases that would be missed in facility-based surveillance systems. Understanding the age distribution, burden, cost-of-illness and transmission of disease is essential to plan interventions and predict their potential impact. The surveillance has been prospectively registered in the Clinical Trial Registry of India ( CTRI/2017/09/009719 ) on 12 September 2017.


Epigenetic patterns newly established after interspecific hybridization in natural populations of Solanum.
Interspecific hybridization is known for triggering genetic and epigenetic changes, such as modifications on DNA methylation patterns and impact on phenotypic plasticity and ecological adaptation. Wild potatoes (Solanum, section Petota) are adapted to multiple habitats along the Andes, and natural hybridizations have proven to be a common feature among species of this group. Solanum × rechei, a recently formed hybrid that grows sympatrically with the parental species S. kurtzianum and S. microdontum, represents an ideal model for studying the ecologically and evolutionary importance of hybridization in generating of epigenetic variability. Genetic and epigenetic variability and their correlation with morphological variation were investigated in wild and ex situ conserved populations of these three wild potato species using amplified fragment length polymorphism (AFLP) and methylation-sensitive amplified polymorphism (MSAP) techniques. We observed that novel methylation patterns doubled the number of novel genetic patterns in the hybrid and that the morphological variability measured on 30 characters had a higher correlation with the epigenetic than with the genetic variability. Statistical comparison of methylation levels suggested that the interspecific hybridization induces genome demethylation in the hybrids. A Bayesian analysis of the genetic data reveled the hybrid nature of S. × rechei, with genotypes displaying high levels of admixture with the parental species, while the epigenetic information assigned S. × rechei to its own cluster with low admixture. These findings suggested that after the hybridization event, a novel epigenetic pattern was rapidly established, which might influence the phenotypic plasticity and adaptation of the hybrid to new environments.


[Localization of NO-synthase in Lugaro cells and mechanisms of NO-ergic interactions between inhibitory interneurons of rabbit cerebellar cortex].
Using histochemical demonstration of NADPH-diaphorase (NADPH-d), several types of NO-producing cells were revealed in rabbit cerebellar cortex. Stellate and basket neurons of molecular layer, Golgi complex and white matter neurons are NADPH positive, while Purkinje cells are negative to NADPH-d. NADPH-d-positive granule cells are found in paraflocculus and flocculus, in vermis and paramedian lobules. They contain predominantly low enzyme activity or are not stained at all. In all the cerebellar cortex areas fusiform Lugaro cells lying under Purkinje cell bodies are stained intensely. Their horizontal dendrites are oriented parallel to Purkinje cells layer. Lugaro cells forming local axonal plexus in lower third of molecular layer and large Lugaro cells with the axon descending to granular layer and basket cell axons form contacts with Lugaro cell dendrites. Clusters of NADPH-d diaphorase positive cells were demonstrated. Dendrosomatic and somatosomatic tangential contacts of Lugaro cells between each other and dendro-dendritic contacts between Lugaro cells and Golgi cells were detected. Total number of Lugaro cells containing NADPH-d in vermis, paramedian lobules, right and left paraflocculus and flocculus equals to 5171. Absence of NO-ergic Lugaro cells in other mammals allows to conclude on the exceptional role of these neurons in rabbit cerebellar cortex.


Pediatric Meckel's Diverticulum: Report of 208 Cases and Review of the Literature.
We report a review of 208 cases of Meckel's diverticulum among pediatric patients from one single institution. One of the aims of this report is to highlight the different diagnostic modalities of Meckel's diverticulum since a majority of cases is undiagnosed prior to surgery. Our review shows 58 cases containing gastric and/or pancreatic heterotopic tissue, including two unique cases. The first case reported is a desmoid tumor arising at the tip of diverticulum, a case that, to our knowledge, has not been previously reported. The second case involves a female patient appearing with an acute abdomen thought to be appendicitis, instead surgery revealed a diverticulum arising from the ileum. The cause of acute abdomen was due to gonococcal infection. In conclusion, we hope that this large series of Meckel's cases will enrich our readers on the differential diagnosis and preoperative diagnostic techniques of Meckels' diverticulum.


The collagenic structure of human digital skin seen by scanning electron microscopy after Ohtani maceration technique.
We performed a morphological scanning electron microscope (SEM) study to describe the fine structure and disposition of collagenous tissue in the human toe. After therapeutic amputation of a human right leg, we applied the Othani maceration technique to the skin of three toes surgically explanted from the foot. We distinguished eight cutaneous regions and focused on some specialized collagenous structures differing in the thickness of the skin. The eight areas investigated were: the dorsal skin, the eponychium, the perionychium, the hyponychium, the region under the visible nail, the nail root, the plantar skin and finally the toe tip. Each of these areas is characterized by a distinctive collagenous surface disposition, with some peculiar features mostly related to dermal papillae. At high magnification, we observed the spatial arrangement of the collagen fibers constituting the top of the dermal papillae that represents the attachment site of the proliferative basal layer of the epidermis. We also noted an impressive density of collagen fibers throughout the thickness of the dermal layer, organized in specialized structures and constituting the skeleton of dermal thermoreceptorial corpuscles or sweat glands. A combination of SEM and Ohtani technique disclosed the three-dimensional architecture of the collagenous matrix of tarsal skin under physiologic conditions, giving a detailed description of the most reactive tissue during pathologic processes.


Role of 18 F-fluoro-2-deoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) in malignant ovarian germ cell tumors: a single-center experience with long term follow-up.
18F-fluoro-2-deoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) is a diagnostic tool widely used in oncology, but to date there are no established recommendations for its use in malignant ovarian germ cell tumors. The aim of this study was to evaluate the role of 18F-FDG PET/CT in the clinical management of patients with malignant ovarian germ cell tumors. This was a retrospective review of 18F-FDG PET/CT scans performed in patients diagnosed with malignant ovarian germ cell tumors treated at the gynecology department of San Gerardo Hospital (Monza, Italy) from June 2006 to December 2016. Data collected included clinical history, radiological, biochemical and pathological evaluation, treatment, follow-up, outcome, and clinical indication for the PET/CT scan. PET/CT findings were categorized as negative/normal (no abnormal FDG uptake or physiological uptake), positive/abnormal (FDG uptake considered to indicate active germ cell malignancy), or equivocal (FDG uptake of uncertain significance, not clearly correlated to neoplastic disease). A total of 69 PET/CT scans in 37 patients were evaluated. The mean age at diagnosis was 25 years (range 20-48). The majority of patients had International Federation of Gynecology and Obstetrics (FIGO) stage I (22/37) disease and had a diagnosis of dysgerminomas (18/37). Imaging indications were initial staging before treatment (4/69, 6%), staging after inadequate staging surgery (24/69, 35%), restaging after adjuvant chemotherapy (17/69, 25%), relapse suspect (9/69, 13%), and follow-up (15/69, 21%). Pathology confirmation of PET/CT results was available in 28/69 (40.5%) studies. All negative PET/CT (15/28) cases were confirmed with laparoscopy as true negative; among 13/28 positive PET cases, histopathology confirmed 7 (54%) as true positive and 6 (46%) as false positive (5 inflammatory and 1 mature teratoma implants). Patient-based analysis showed 100% sensitivity, 71% specificity, 54% positive predictive value, 100% negative predictive value, and 79% accuracy. Clinical follow-up was available in 41 (59.4%) of 69 PET/CT images: 28/41 studies were negative and 13/41 positive. A mean follow-up of 28 months (median 15, range 5-102) confirmed negative PET/CT studies. A total of 13 positive PET/CT patients underwent chemotherapy with subsequent evidence of disease response. PET/CT in malignant ovarian germ cell tumors was mainly performed for staging after inadequate staging surgery or for restaging after adjuvant chemotherapy. PET/CT was associated with high sensitivity and negative predictive value.


Porous Co3O4 nanowires derived from long Co(CO3)(0.5)(OH)·0.11H2O nanowires with improved supercapacitive properties.
Porous Co(3)O(4) nanowires with large aspect ratio have been obtained by annealing long Co(CO(3))(0.5)(OH)·0.11H(2)O precursor nanowires synthesized by a facile hydrothermal method. The results show that the amount of the additive (urea) has an important impact on the morphology of the as-synthesized cobalt-carbonate-hydroxide intermediate, where the uniformity and the overall structure can be controlled by changing the urea concentration. After the heat treatment, the as-obtained phase-pure Co(3)O(4) nanowires with a well retained structure are applied as the electrode material for supercapacitors, and the sample exhibits excellent performance with a high specific capacitance of 240 F g(-1) after 2000 charge/discharge cycles, corresponding to a retention of 98% of the initial capacitance.


Cloning of determinants encoding F165(2) fimbriae from porcine septicaemic Escherichia coli confirms their identity as F1C fimbriae.
Cloning of the f165(2) operon that encodes F1C-like fimbriae in Escherichia coli indicates that this operon is a member of the S/Foc family. The genetic determinant coding for the F165(2) fimbriae was cloned from the chromosome of the porcine E. coli wild-type strain 4787 (O115:K-1:H51:F165). The cloned F165(2) and the wild-type operon expressed a major fimbrial protein subunit of molecular mass 17.2 kDa that was detected by anti-F165 and anti-F1C polyclonal sera. The sequences of the f165(2)A and f165(2)FGH genes are reported. Major subunit gene f165(2)A encodes a mature protein of 156 amino acids. Minor subunit genes F, G and H encode mature proteins of 148, 145 and 276 amino acids, respectively. The amino acid sequences of the four proteins share similarities with those of the known S and F1C fimbrial antigens that are produced by extraintestinal E. coli which is associated with sepsis, urinary tract infections and newborn meningitis. The F165(2)A protein was identical to the major subunit protein of F1C, with a difference only at the first position. It was also similar, to a lesser extent, to the major subunit proteins of SfaI and SfaII fimbriae. F165(2)F was identical to FocF and SfaG/SfaIIS, and F165(2)H was more closely related to FocG than to SfaIS/SfaIIS, and F165(2)H was more closely related to FocH than to SfaIH/SfaIIH.


Genetic variation, classification and 'race'.
New genetic data has enabled scientists to re-examine the relationship between human genetic variation and 'race'. We review the results of genetic analyses that show that human genetic variation is geographically structured, in accord with historical patterns of gene flow and genetic drift. Analysis of many loci now yields reasonably accurate estimates of genetic similarity among individuals, rather than populations. Clustering of individuals is correlated with geographic origin or ancestry. These clusters are also correlated with some traditional concepts of race, but the correlations are imperfect because genetic variation tends to be distributed in a continuous, overlapping fashion among populations. Therefore, ancestry, or even race, may in some cases prove useful in the biomedical setting, but direct assessment of disease-related genetic variation will ultimately yield more accurate and beneficial information.


Baseline and Clinical Factors Associated with Response to Amblyopia Treatment in a Randomized Clinical Trial.
We sought to identify baseline and clinical factors that were predictive of the response to amblyopia treatment. We report that binocular amblyopia treatment may be especially effective for moderate amblyopia in orthotropic children. We previously reported results from the primary cohort (n = 28) enrolled in a randomized clinical trial (NCT02365090), which found that binocular amblyopia treatment was more effective than patching. Enrollment of an additional 20 children was pre-planned to provide the opportunity to examine factors that may be predictive of response to amblyopia treatment. Forty-eight children (4 to 10 years old) were enrolled, with 24 randomized to contrast-rebalanced binocular game treatment (1 hour a day, 5 days a week) and 24 to patching treatment (2 hours a day, 7 days a week). The primary outcome was change in amblyopic eye best-corrected visual acuity at the 2-week visit. Baseline factors examined were age at enrollment, visual acuity, stereoacuity, and suppression. Clinical factors were etiology, age at diagnosis, prior treatment, and ocular alignment. At 2 weeks, visual acuity improvement was significantly greater with the binocular game than patching. Children with moderate amblyopia and orthotropia had more visual acuity improvement with binocular game play than did those with severe amblyopia. In addition, children who spent more time playing the binocular game had more improvement. We were not able to confidently identify any baseline or clinical factors that were associated with response to patching treatment. Binocular amblyopia treatment was more effective among orthotropic children with moderate amblyopia than among children with microtropia or severe amblyopia.


Symptom reduction and suicide risk in patients treated with placebo in antidepressant clinical trials: a replication analysis of the Food and Drug Administration Database.
The assumption that depressed patients who are assigned to placebo in antidepressant clinical trials are exposed to substantial morbidity and mortality has not been based on research data. Because of worldwide concern about placebo use and the implications of our earlier findings of no increased suicide risk in placebo-treated patients, we conducted a replication study in a new patient sample. We assessed suicide risk and symptom reduction among placebo-treated patients participating in antidepressant clinical trials for two recently approved antidepressants, venlafaxine ER and citalopram, which were unavailable during our previous study. Among 23,201 participant patients, 32 committed suicide and 172 attempted suicide. Rates of suicide and attempted suicide did not differ significantly among the placebo- and drug-treated groups. Based on patient exposure years, annual rates of suicide and attempted suicide were 0.5 and 6.7% with placebo, 0.9% with active comparator (rates for attempted suicide are unavailable), and 0.6 and 6.3% with investigational antidepressants. Symptom reduction was 47.9% with investigational drugs (n = 1172), 47.5% with active comparators (n = 161), and 35.5% with placebo (n = 606). These data may inform discussions about the use of placebo in antidepressant clinical trials.


Guidelines for training in Cardiovascular Magnetic Resonance (CMR).
These "Guidelines for Training in Cardiovascular Magnetic Resonance" were developed by the Clinical Practice Committee of the Society for Cardiovascular Magnetic Resonance (SCMR) and approved by the SCMR Board of Trustees.


Early Detection of Severe Functional Impairment Among Adolescents With Major Depression Using Logistic Classifier.
Machine learning is about finding patterns and making predictions from raw data. In this study, we aimed to achieve two goals by utilizing the modern logistic regression model as a statistical tool and classifier. First, we analyzed the associations between Major Depressive Episode with Severe Impairment (MDESI) in adolescents with a list of broadly defined sociodemographic characteristics. Using findings from the logistic model, the second and ultimate goal was to identify the potential MDESI cases using a logistic model as a classifier (i.e., a predictive mechanism). Data on adolescents aged 12-17 years who participated in the National Survey on Drug Use and Health (NSDUH), 2011-2017, were pooled and analyzed. The logistic regression model revealed that compared with males and adolescents aged 12-13, females and those in the age groups of 14-15 and 16-17 had higher risk of MDESI. Blacks and Asians had lower risk of MDESI than Whites. Living in single-parent household, having less authoritative parents, having negative school experiences further increased adolescents' risk of having MDESI. The predictive model successfully identified 66% of the MDESI cases (recall rate) and accurately identified 72% of the MDESI and MDESI-free cases (accuracy rate) in the training data set. The rates of both recall and accuracy remained about the same (66 and 72%) using the test data. Results from this study confirmed that the logistic model, when used as a classifier, can identify potential cases of MDESI in adolescents with acceptable recall and reasonable accuracy rates. The algorithmic identification of adolescents at risk for depression may improve prevention and intervention.


Chlorinated paraffins and dechloranes in free-range chicken eggs and soil around waste disposal sites in Tanzania.
Electronic waste is a source of both legacy and emerging flame retardants to the environment, especially in regions where sufficient waste handling systems are lacking. In the present study, we quantified the occurrence of short- and medium chain chlorinated paraffins (SCCPs and MCCPs) and dechloranes in household chicken (Gallus domesticus) eggs and soil collected near waste disposal sites on Zanzibar and the Tanzanian mainland. Sampling locations included an e-waste facility and the active dumpsite of Dar es Salaam, a historical dumpsite in Dar es Salaam, and an informal dumpsite on Zanzibar. We compared concentrations and contaminant profiles between soil and eggs, as free-range chickens ingest a considerable amount of soil during foraging, with potential for maternal transfer to the eggs. We found no correlation between soil and egg concentrations or patterns of dechloranes or CPs. CPs with shorter chain lengths and higher chlorination degree were associated with soil, while longer chain lengths and lower chlorination degree were associated with eggs. MCCPs dominated the CP profile in eggs, with median concentrations ranging from 500 to 900 ng/g lipid weight (lw) among locations. SCCP concentrations in eggs ranged from below the detection limit (LOD) to 370 ng/g lw. Dechlorane Plus was the dominating dechlorane compound in all egg samples, with median concentrations ranging from 0.5 to 2.8 ng/g lw. SCCPs dominated in the soil samples (400-21300 ng/g soil organic matter, SOM), except at the official dumpsite where MCCPs were highest (65000 ng/g SOM). Concentrations of dechloranes in soil ranged from below LOD to 240 ng/g SOM, and the dominating compounds were Dechlorane Plus and Dechlorane 603. Risk assessment of CP levels gave margins of exposure (MOE) close to or below 1000 for SCCPs at one location.


Reliability of goniometry-based Q-angle.
To establish the stability reliability, precision, and minimum value for detecting real differences for quadriceps angle (Q-angle) measurements based on standardized protocols and surface goniometry. An intratester reliability study. University research laboratory. Fifty-two healthy, young, relatively lean adults (25 men and 27 women) from the university community with no history of knee injury. Q-angle was assessed with surface goniometry on 2 separate occasions separated by 48 hours. Subjects assumed a supine position with: (1) extended hips and knees, (2) neutral hip rotational position, (3) neutral foot position, and (4) isometrically contracted quadriceps femoris muscles. The axis of a manual extendable-arm goniometer was placed over the center of the right patella with the proximal arm situated over the anterior-superior iliac spine and the distal arm over the center of the tibial tuberosity. Stability reliability was calculated with use of intraclass correlation (ICC, 2-way random model) and precision was calculated by standard error of measurement (SEM). The 95% limits of agreement also were calculated to estimate the minimum detectable difference in Q-angles. The lowest acceptable ICC was set at ≥0.70 for stability reliability. The following ICC (SEM) values were found: all subjects = 0.88 (1.0°), men = 0.77 (1.0°), and women = 0.85 (1.0°). The 95% limits of agreement were 3° for the same 3 groups. The surface goniometry protocol described herein appeared to be reliable for relatively lean young men and women. Although measures were precise to 1.0°, it appears a difference of 3° may be needed to detect a real difference in Q-angles when measured in this fashion.


Expression and function of CD40 on Hodgkin and Reed-Sternberg cells and the possible relevance for Hodgkin's disease.
CD40 was originally described as a B-cell-restricted antigen and was subsequently found to be a member of the tumor necrosis factor (TNF) receptor superfamily. CD40 is also expressed on dendritic cells, thymic epithelium, monocytes, and some carcinoma cell lines, and plays a critical role in cell contact-dependent activation. Primary and cultured Hodgkin and Reed-Sternberg (H-RS) cells, the presumed malignant cells of Hodgkin's disease (HD); were found to express high levels of cell surface CD40. We found that recombinant CD40 ligand (CD40L) induced interleukin-8 (IL-8) secretion and enhanced IL-6, TNF, and lymphotoxin-alpha (LT-alpha/TNF-beta) release from cultured H-RS cells. These cytokines play a significant role in the clinical presentation and pathology of HD, a tumor of cytokine-producing cells. CD40L had no mitogenic activity for HD-derived cell lines. In contrast, CD40L enhanced expression of costimulatory molecules intracellular adhesion molecule-T and B7-1 on cultured H-RS cells, both of which are overexpressed on primary H-RS cells. In addition, CD40L induced a 40% to 60% reduction of the expression of the HD-associated CD30 antigen, another member of the TNF receptor superfamily. Primary and cultured H-RS cells express not only CD30, but also CD40. CD40L has pleiotropic biologic activities on H-RS cells, and the CD40-CD40L interaction might be a critical element in the deregulated cytokine network and cell contact-dependent activation cascade typical for HD.


Expressive drawing ability in children with autism.
The autistic impairments in emotional and social competence, imagination and generating ideas predict qualitative differences in expressive drawings by children with autism beyond that accounted by any general learning difficulties. In a sample of 60 5-19-year-olds, happy and sad drawings were requested from 15 participants with non-savant autism and compared with those drawn by three control groups matched on either degree of learning difficulty (MLD), mental age (MA) or chronological age (CA). All drawings were rated by two artists on a 7-point quality of expression scale. Contrary to our predictions, the drawings from the autistic group were rated similar to those of the MA and MLD groups. Analysis of the people and social content of the drawings revealed that although children with autism did not draw fewer people, they did draw more immature forms than mental age controls. Furthermore, there was tentative evidence that fewer social scenes were produced by the autism sample. We conclude that the overall merit of expressive drawing in autism is commensurate with their general learning difficulties, but the social/emotional impairment in autism affects their drawings of people and social scenes.