Interdependence of pyrene interactions and tetramolecular G4-DNA assembly.
Controlling the arrangement of organic chromophores in supramolecular architectures is of primary importance for the development of novel functional molecules. Insertion of a twisted intercalating nucleic acid (TINA) moiety, containing phenylethynylpyren-1-yl derivatives, into a G-rich DNA sequence alters G-quadruplex folding, resulting in supramolecular structures with defined pyrene arrangements. Based on CD, NMR and ESI-mass-spectra, as well as TINA excited dimer (excimer) fluorescence emission we propose that insertion of the TINA monomer in the middle of a dTG4T sequence (i.e. dTGGXGGT, where X is TINA) converts a parallel tetramolecular G-quadruplex into an assembly composed of two identical antiparallel G-quadruplex subunits stacked via TINA-TINA interface. Kinetic analysis showed that TINA-TINA association controls complex formation in the presence of Na(+) but barely competes with guanine-mediated association in K(+) or in the sequence with the longer G-run (dTGGGXGGGT). These results demonstrate new perspectives in the design of molecular entities that can kinetically control G-quadruplex formation and show how tetramolecular G-quadruplexes can be used as a tuneable scaffold to control the arrangement of organic chromophores.


Differential roles for Sox15 and Sox2 in transcriptional control in mouse embryonic stem cells.
Sox family transcription factors play essential roles in cell differentiation, development, and sex determination. Sox2 was previously thought to be the sole Sox protein expressed in mouse embryonic stem (ES) cells. Sox2 associates with Oct3/4 to maintain self-renewal of ES cells. In the current study, digital differential display identified transcripts for an additional Sox family member, Sox15, enriched in mouse ES cells. Reverse transcription-PCR confirmed that Sox15 expression is highest in undifferentiated ES cells and repressed upon differentiation. Sox15 is expressed at low levels in several tissues, including testis and muscle. In vitro studies showed that Sox15, like Sox2, associated with Oct3/4 on DNA sequences containing the octamer motif and Sox-binding site. Gel mobility shift assays and SELEX analyses showed that Sox15 binds similar DNA sequences as Sox2 but with weaker affinity. In contrast to the early embryonic lethality observed in Sox2-null mice, Sox15-null ES cells and mice were grossly normal. DNA microarray analyses revealed that Otx2, Ctgf, Ebaf, and Hrc are dysregulated in Sox15-null ES cells, however. Chromatin immunoprecipitation showed that Sox15, but not Sox2, bound to a Sox consensus binding site within the Hrc gene. Taken together, these data demonstrate differential roles for Sox15 and Sox2 in transcriptional control in mouse ES cells.


Specialised fetal and maternal service: outcome of pre-gestational diabetes.
The objective of the study was to determine the outcome of pregnancies with pre-gestational diabetes mellitus (PGDM) in the presence of a specialised maternal and fetal service. Prospective data included mothers with documented pre-gestational diabetes (PGDM) delivered between 1 January 2007 and 31 December 2009. A total of 138 patients with PGDM were included in this study. The post-lunch glucose level at 34 weeks was significantly lower than at 30 weeks' gestation (p =0.007) and 37 weeks' gestation (p =0.02). No correlation was observed between maternal blood sugar and birth weight. The incidence of pre-term labour, stillbirth and admission to the NICU was similar to the control group. Caesarean section rate was 39.1%, and the main indication was previous caesarean section. The incidence of fetal anomalies was significantly higher than in the control group. It was concluded that the presence of specialised maternal and fetal clinics reduces complications related to prenatal glycaemic control. However, complications related to preconception care remains high.


Adhesion G Protein-Coupled Receptor G2 Promotes Hepatocellular Carcinoma Progression and Serves as a Neutrophil-Related Prognostic Biomarker.
Adhesion G protein-coupled receptor G2 (ADGRG2) is an orphan adhesion G protein-coupled receptor (GPCR), which performs a tumor-promoting role in certain cancers; however, it has not been systematically investigated in hepatocellular carcinoma (HCC). In the current study, we utilized multiple databases to analyze the expression and diagnostic and prognostic value of ADGRG2 in HCC and its correlation with immune infiltration and inflammatory factors. The function and upstream regulatory miRNA of ADGRG2 were validated through qPCR, Western blot, CCK8, wound healing, and dual luciferase assays. It turned out that ADGRG2 was significantly higher in HCC and had a poor survival rate, especially in AFP ≤ 400 ng/mL subgroups. Functional enrichment analysis suggested that ADGRG2 may be involved in cancer pathways and immune-related pathways. In vitro, siRNA-mediated ADGRG2 silencing could inhibit the proliferation and migration of Huh7 and HepG2 cells. There was a highly significant positive correlation between ADGRG2 and neutrophils. Moreover, NET-related genes were filtered and confirmed, such as ENO1 and S100A9. Meanwhile, the high expression of ADGRG2 was also accompanied by the highest number of inflammatory cytokines, chemokines, and chemokine receptors and good immunotherapy efficacy. Finally, AGDGR2 may be sensitive to two drugs (PIK-93 and NPK76-II-72-1) and can be targeted by miR-326. In conclusion, ADGRG2 may serve as a novel biomarker and drug target for HCC diagnosis, immunotherapy, and prognosis and was related to neutrophils and the inflammatory process of liver cancer development.


Correlation between fitness and fatness in 6-14-year old Serbian school children.
Lack of physical activity and/or physical fitness are some reasons epidemiologists suggest for increase in childhood obesity in the last 20 years, with clear correlation between body composition and physical activity and/or physical fitness yet to be determined. The objectives of the study were to (a) investigate the prevalence of overweight and obesity among Serbian school children and (b) determine the relationship between indicators of physical activity and body fatness in Serbian school children aged 6-14 years. The study subjects included a representative sample of Serbian elementary school children (n = 1,121-754 boys and 367 girls-aged 6.2-14.1 years), all of whom were recruited in the OLIMP (Obesity and Physical Activity among Serbian School Children) study. Anthropometric and physical fitness values, including body mass index (BMI), waist-circumference, body-fat, and aerobic capacity, were measured in all the children. Significant differences were found between male and female children regarding the prevalence of obesity (6.8% vs 8.2%, p < 0.05, boys and girls respectively). Boys had significantly lower body mass, BMI, waist-circumference, sum of six skinfolds, and body-fat compared to their female counterparts (p < 0.05). The highest level of weight, BMI, body-fat, and waist-circumference observed in a 14-year old girl (96.3 kg, 40.5 kg/m2, 54.5%, 91.4 cm respectively) implies the existence of extreme obesity in Serbian school children. The negative relationship between body-fat and maximal oxygen (VO2max) uptake was moderately high (r = -0.76; p < 0.05). The study has shown a high prevalence of adiposity among Serbian school children, with a strong negative relationship between aerobic fitness and body fatness. Data of the study emphasize the necessity to identify children with weight problems and to develop early interventions to improve physical activity in children and prevent the increase of childhood obesity.


The discriminatory zone of beta-hCG for vaginal probes.
Recent developments in both laboratory measurements and ultrasound technology have revolutionized the management of early pregnancy. The discriminatory zone concept is a direct result of these developments. By correlating the serum beta-hCG values to the size of an intrauterine gestational sac, a value can be chosen that corresponds to the threshold of visualization of the sac. If the beta-hCG is above this value, a sac must be seen, and if it is not, aggressive steps should be taken to determine whether the pregnancy is abnormal or ectopic. The discriminatory zone may vary among institutions due to different equipment and assays. Thus, its value should be calculated individually at each institution. A proper discriminatory zone and a management protocol such as the one above can eliminate much of the uncertainty in the management of suspected ectopic and early pregnancies.


Sanctification and spiritual disclosure in parent-child relationships: implications for family relationship quality.
Social scientific research on family life, religion, and spirituality tends to focus on global religiousness and spirituality with few studies seeking to understand interpersonal religious and spiritual contributors, namely sanctification and spiritual disclosure, from multiple family members' perspectives. This study explored 91 mother-college student and 64 father-college student dyads who rated their use of spiritual disclosure and theistic and nontheistic sanctification of the parent-child dyad in relation to parent-child relationship quality (e.g., parent-child relationship satisfaction and open communication). Results indicate significant positive links between higher levels of spiritual disclosure and greater theistic and nontheistic sanctification, for mothers, fathers, and their children. However, only greater nontheistic sanctification and higher levels spiritual disclosure were significantly related to increased parent-child relationship quality. Through use of Actor-Partner Interdependence Models (APIMs) results indicated unique contributions of spiritual disclosure to parent-child relationship quality above nontheistic sanctification for open communication in the family. However, full models, which included nontheistic sanctification and spiritual disclosure, predict college students' relationship satisfaction with their mothers and fathers. Implications for interpersonal religiousness and spirituality as contributors to familial relationship quality in research and practice are discussed.


WITHDRAWN-Administrative Duplicate Publication: Voice Quality in Adults Treated for Unilateral Cleft Lip and Palate: Long-term Follow-up After 1- or 2-Stage Palate Repair.
Morén, S., Lindestad, P. Å., Holmström, M., & Mani, M. (2018). Voice Quality in Adults Treated for Unilateral Cleft Lip and Palate: Long-term Follow-up After 1- or 2-Stage Palate Repair. The Cleft Palate-Craniofacial Journal, 55(5), 758-768. DOI: 10.1177/1055665618754946 Article withdrawn by publisher. Due to an administrative error, this article was accidentally published in Volume 55 Issue 5 as well as Volume 55 Issue 8 of publishing year 2018 with different DOIs and different page numbers. The incorrect version of the article with DOI: 10.1177/1055665618754946 has been replaced with this correction notice. The correct and citable version of the article remains: Morén, S., Lindestad, P. Å., Holmström, M., & Mani, M. (2018). Voice Quality in Adults Treated for Unilateral Cleft Lip and Palate: Long-Term Follow-Up After One- or Two-Stage Palate Repair. The Cleft Palate-Craniofacial Journal, 55(8), 1103-1114. DOI: 10.1177/1055665618764521.


Hyaluronic acid and thrombin upregulate MT1-MMP through PI3K and Rac-1 signaling and prime the homing-related responses of cord blood hematopoietic stem/progenitor cells.
One of the hurdles of cord blood (CB) transplantation is delayed hematopoietic engraftment. Previously, we demonstrated that supernatants isolated from leukapheresis products of granulocyte-colony stimulating factor (G-CSF)-mobilized patients primed the homing of hematopoietic stem/progenitor cells (HSPC) by enhancing their chemotactic responses to stromal cell-derived factor (SDF)-1 and stimulating matrix metalloproteinases (MMPs) MMP-2 and MMP-9. Since membrane type 1 (MT1)-MMP activates proMMP-2 and localizes proteolytic activity at the leading edge of migrating cells, in this study we investigated whether MT1-MMP contributes to the priming of the homing-related responses of CB HSPC. We found that components of supernatants of leukapheresis products such as hyaluronic acid and thrombin (i) increase the secretion of proMMP-9 and transcription and protein synthesis of MT1-MMP in CB CD34(+) cells; (ii) increase the levels of active MMP-2 in co-cultures of CD34(+) cells with endothelial cells; (iii) increase the chemoinvasion across reconstituted basement membrane Matrigel of CD34(+) cells toward a low SDF-1 gradient (20 ng/mL); and (iv) activate mitogen-activated protein kinase, phosphatidylinositol 3-kinase, and Rac-1 signaling pathways. Inhibition of phosphatidylinositol 3-kinase and Rac-1 by their respective inhibitors LY290042 and NSC23766 attenuated MT1-MMP expression in CB CD34(+) cells, leading to reduced proMMP-2 activation and HSPC trans-Matrigel chemoinvasion toward SDF-1. Thus, our data suggest that MT1-MMP plays an important role in the homing-related responses of HSPC, and we propose that pretreatment of CB HSPC with hyaluronic acid or thrombin before transplantation could improve their homing and engraftment.


Middle-term effects of education programme in chronic low back pain patients to an adherence to physical activity: A randomized controlled trial.
This study was to measure the impact of an education programme in Chronic Low Back Pain (CLBP) patients on their changing behaviour; particularly on the resumption or maintenance of physical activity (PA) at three and six months after care. A two-group, randomised controlled clinical trial was conducted on 68 patients as the control group (CG) and 68 as the experimental group (EG). All patients benefited from a four-week multidisciplinary programme, with an educative programme for the EG. The dropouts of patients during the follow-up period and the PA level were measured at three and six months after the end of care. At 3-months, the EG presented a significant higher level of physical activity (91% of EG versus 77% of CG, p < 0.001) and at six months, this group presented a lower dropout rate than CG (60% of EG versus 73% of CG, p = 0.017). The educative programme, added to a multidisciplinary care plan, can improve adherence to the care programme and the continuum of PA. Add an educative programme focused on motivation, physical activity, stress management and daily activities seems necessary to accompany CLBP patient towards the maintenance of a regular PA.


Saccadic performance characteristics and the behavioural neurology of Tourette's syndrome.
To better understand the neuropathological correlates of Tourette's syndrome (TS), measures of saccadic eye movement performance were examined among patients with TS. A case-control design was used. Twenty one patients with DSM-IV TS (mean age 40.6 years (SD 11.0); 38% female) mainly recruited from UCSD Psychiatry Services, and a community based sample of 21 normal subjects (mean age 34.6 years (SD 13.4); 43% women) participated in this study. Participants were administered ocular motor tasks assessing visual fixation, and the generation of prosaccades, predictive saccades, and antisaccades. Saccadic reaction time, amplitude, duration, and mean and peak velocity were computed. Intrusive saccades during visual fixation and the proportion of correct antisaccade responses were also evaluated. The groups had similar visual fixation performance. Whereas patients with TS generated prosaccades with normal reaction times and amplitudes, their saccade durations were shorter and their mean velocities were higher than in normal subjects. During a prosaccade gap task, patients with TS exhibited an increased proportion of anticipatory saccades (RTs<90). The proportion of "express" saccades (90<RTs<135) did not differ between groups. Patients with TS had fewer correct antisaccade responses than did normal subjects, an effect accounted for by 19% of the patients. Antisaccade reaction times among patients with TS were increased during an overlap version of the task. These findings suggest that TS mildly affects the ocular motor control circuitry associated with saccade inhibition.


High-resolution computed tomography in silicosis: correlation with chest radiography and pulmonary function tests.
To correlate tomographic findings with pulmonary function findings, as well as to compare chest X-ray findings with high resolution computed tomography (HRCT) findings, in patients with silicosis. A cross-sectional study was conducted in 44 non-smoking patients without a history of tuberculosis. Chest X-ray findings were classified according to the International Labour Organization recommendations. Using a semiquantitative system, the following HRCT findings were measured: the full extent of pulmonary involvement; parenchymal opacities; and emphysema. Spirometry and forced oscillation were performed. Pulmonary volumes were evaluated using the helium dilution method, and diffusing capacity of the lung for carbon monoxide (DLCO) was assessed. Of the 44 patients studied, 41 were male. The mean age was 48.4 years. There were 4 patients who were classified as category 0 based on X-ray findings and as category 1 based on HRCT findings. Using HRCT scans, we identified progressive massive fibrosis in 33 patients, compared with only 23 patients when X-rays were used. Opacity score was found to correlate most closely with airflow, DLCO and compliance. Emphysema score correlated inversely with volume, DLCO and airflow. In this sample of patients presenting a predominance of large opacities (75% of the individuals), the deterioration of pulmonary function was associated with the extent of structural changes. In the early detection of silicosis and the identification of progressive massive fibrosis, HRCT scans are superior to X-rays.


Differential action of polycyclic aromatic hydrocarbons on endogenous estrogen-responsive genes and on a transfected estrogen-responsive reporter in MCF-7 cells.
Polycyclic aromatic hydrocarbons (PAHs) are common environmental pollutants that have been extensively studied for multiple toxicological endpoints in both laboratory animals and humans. The purpose of this study was to investigate the estrogenicity of PAHs in the human breast cancer cell line MCF-7. We investigated 14 PAHs for their ability to bind either the estrogen receptor (ER) or the aryl hydrocarbon receptor (AhR) and to activate target gene expression. PAHs were tested in a human recombinant estrogen receptor (hrER) competitive binding assay, and in both an estrogen response element (ERE)- and xenobiotic response element (XRE)-mediated reporter gene assay. We used quantitative RT-PCR to examine selected PAHs that showed activity in the ERE reporter gene assay for their ability to upregulate estrogen-responsive genes HEM45, progesterone receptor, and pS2, and the aryl hydrocarbon-responsive CYP1A1 gene. None of the 14 PAHs bound the hrER, but five of the PAHs (anthracene, B[a]A, chrysene, B[b]F, and B[a]P) induced ER-reporter activity. This activity was dependent on the metabolism of PAHs in MCF-7 cells via the AhR pathway, which resulted in the formation of metabolites that bound the ER. None of the five PAHs that induced the ER-reporter were found to upregulate estrogen-responsive genes, yet four of the five PAHs induced AhR-dependent CYP1A1 gene expression. In contrast, a metabolite of B[a]P, 3'OH-B[a]P, and a PCB metabolite, 4'OH-2,4,6-BP, did weakly upregulate all three estrogen-responsive genes. Data from these studies indicate that induction of ER-reporter activity alone does not necessarily parallel endogenous gene transcription, and that the reporter gene assay may detect interactions that are not functional in vivo.


A double-blind randomised clinical trial of two techniques for gingival displacement.
Knowledge about security and the potential damage originated by the gingival displacement techniques has not been described through randomised clinical studies. This crossover, double-blind, randomised clinical trial evaluated clinical and immunological factors related to conventional and cordless gingival displacement (GD) techniques, and patients' perceptions in 12 subjects with the employment of 2 GD techniques: conventional (gingival cord + 25% AlCl3 astringent gel) and cordless (15% AlCl3 astringent-based paste). In each subject, two anterior teeth were treated and a 10-day wash-out period separated the two treatments. Periodontal indices were evaluated before (baseline) and 1 and 10 days after GD. Interleukin 1β, interleukin 6 and tumour necrosis factor α concentrations in gingival crevicular fluid were measured before and 1 day after GD. Subjective parameters (pain, unpleasant taste and stress) were also evaluated. Data were analysed by one-way repeated-measures analysis of variance and Tukey's test (immunological factors), the Friedman test (periodontal parameters) and Fisher's exact or chi-squared test (subjective parameters), with a significance level of 95%. Gingival bleeding index, probing depth and plaque index values did not differ significantly between groups at any timepoint. Neither technique resulted in worse periodontal indices. Both techniques yielded similar results for pain and unpleasant taste, but conventional GD was significantly more stressful than cordless GD for volunteers. Both treatments significantly increased mean concentrations of the three cytokines, with the conventional technique producing the highest cytokine levels. Cordless GD is less stressful for patients and results in lower post-treatment levels of inflammatory cytokines compared with conventional GD.


Production of a chimeric form of CD23 that is oligomeric and blocks IgE binding to the Fc epsilonRI.
The low affinity receptor for IgE (Fc epsilonRII/CD23) has previously been shown to interact with IgE with a dual affinity. Three chimeric constructs were created containing the lectin domain (amino acids 172-188) or the "neck" and lectin domain (amino acids 157-188) attached to subunits of oligomeric proteins. All chimeras were incapable of interacting with IgE with either a high or low affinity, indicating that the alpha-helical stalk of CD23 is important for orienting the lectin heads such that an interaction with IgE can occur. This concept received further support in that a chimeric CD23 composed of the human CD23 stalk and the mouse CD23 lectin head bound mouse IgE with a dual affinity, but could only bind rat IgE with a low affinity. Effort was next concentrated on a construct consisting of the entire extracellular (EC) region of CD23. A mutation to the first cleavage site of CD23 (C1M) resulted in a more stable molecule as determined by a decrease of soluble CD23 release. A soluble chimeric EC-C1M was prepared by attaching an isoleucine zipper to the amino terminus (lzEC-C1M). The interaction with IgE by lzEC-C1M was found to be superior to that seen with EC-CD23. The lzEC-C1M could inhibit binding of IgE to both CD23 and the high affinity receptor for IgE, Fc epsilonRI, providing further evidence for a strong interaction with IgE. Fc epsilonRI inhibition (approximately 70%) was seen at equimolar concentrations of lzEC-C1M, implying the effectiveness of this chimera and suggesting its potential therapeutic value.


The fibronectin-binding capacity and host cell adherence of Streptococcus pyogenes strains are discordant with each other.
Surface exposed fibronectin-binding proteins (FBPs) play an important role in the adherence of Streptococcus pyogenes (group A streptococcus, GAS) to host cells. This pathogen expresses numerous FBPs, of which SfbI, SfbII and PrtF2 are major surface exposed FBPs. However, GAS strains differ in the genetic potential to express these proteins. To test whether this difference reflects in differences in fibronectin (Fn) binding, a set of circulating strains previously examined for adherence to host cells was used. The 68 distinct strains were isolated from throat, skin and blood. They were analyzed for (a) the presence of genes for SfbI, SfbII and PrtF2 and (b) the extent of Fn binding. The results suggest that strains possessing two or more of the genes for these FBPs bound Fn significantly more than strains possessing none or one of the genes. No correlation between the extent of Fn binding and the tissue site of isolation was found. Furthermore, together with our previous studies on adherence capacity of these GAS strains, we found no correlation between Fn binding ability and the avidity of the strains to adhere to epithelial cells. We suggest that while Fn binding is important for adhesion, for many GAS strains the extent of Fn binding is not the critical determinant of adherence.


CD34(+) acute myeloid and lymphoid leukemic blasts can be induced to differentiate into dendritic cells.
CD34(+) hematopoietic stem cells from normal individuals and from patients with chronic myelogenous leukemia can be induced to differentiate into dendritic cells (DC). The aim of the current study was to determine whether acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) cells could be induced to differentiate into DC. CD34(+) AML-M2 cells with chromosome 7 monosomy were cultured in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor alpha (TNFalpha), and interleukin-4 (IL-4). After 3 weeks of culture, 35% of the AML-M2 cells showed DC morphology and phenotype. The DC phenotype was defined as upmodulation of the costimulatory molecules CD80 and CD86 and the expression of CD1a or CD83. The leukemic nature of the DC was validated by detection of chromosome 7 monosomy in sorted DC populations by fluorescence in situ hybridization (FISH). CD34(+) leukemic cells from 2 B-ALL patients with the Philadelphia chromosome were similarly cultured, but in the presence of CD40-ligand and IL-4. After 4 days of culture, more than 58% of the ALL cells showed DC morphology and phenotype. The leukemic nature of the DC was validated by detection of the bcr-abl fusion gene in sorted DC populations by FISH. In functional studies, the leukemic DC were highly superior to the parental leukemic blasts for inducing allogeneic T-cell responses. Thus, CD34(+) AML and ALL cells can be induced to differentiate into leukemic DC with morphologic, phenotypic, and functional similarities to normal DC.


Nitric oxide-mediated inhibition of NFkappaB regulates hyperthermia-induced apoptosis.
Ascertaining the upstream regulatory mechanisms of hyperthermia-induced apoptosis is important to understand the role of hyperthermia in combined modality cancer therapy. Accordingly, we investigated whether (i) hyperthermia-induced apoptosis is mediated through the nitric oxide (NO) signaling pathway and (ii) inhibition of post-translational modification of IkappaBalpha and down regulation of NFkappaB-DNA binding activity is an intermediate step in NO-dependent apoptosis in MCF-7 breast cancer cells. For hyperthermia treatment, the cells were exposed to 43 degrees C. Intracellular NO levels measured by the fluorescent intensity of DAF-2A and iNOS expression by immunobloting revealed an increased level of iNOS dependent NO production after 43 degrees C. Apoptosis measured by Annexin V expression and cell survival by clonogenic assay showed a 20% increase in apoptosis after 43 degrees C treatments. EMSA analysis showed a dose-dependent inhibition of NFkappaB-DNA binding activity. The hyperthermia-mediated inhibition of NFkappaB was persistent even after 48 h. Inhibition of NO by L-NAME rescued the NFkappaB-DNA binding activity and inhibits heat-induced apoptosis. Similarly, over-expression of NFkappaB by transient transfection inhibits heat-induced apoptosis. These results demonstrate that apoptosis upon hyperthermia exposure of MCF-7 cells is regulated by NO-mediated suppression of NFkappaB.


Are school-based physical activity interventions effective and equitable? A meta-analysis of cluster randomized controlled trials with accelerometer-assessed activity.
The prevalence of childhood obesity is increasing at epidemic rates globally, with widening inequalities between advantaged and disadvantaged groups. Despite the promise of schools as a universal context to access and influence all children, the potential of school-based interventions to positively impact children's physical activity behaviour, and obesity risk, remains uncertain. We searched six electronic databases to February 2017 for cluster randomized trials of school-based physical activity interventions. Following data extraction, authors were sent re-analysis requests. For each trial, a mean change score from baseline to follow-up was calculated for daily minutes of accelerometer-assessed moderate-to-vigorous physical activity (MVPA), for the main effect, by gender, and by socio-economic position (SEP). Twenty-five trials met the inclusion criteria; 17 trials provided relevant data for inclusion in the meta-analyses. The pooled main effect for daily minutes of MVPA was nonexistent and nonsignificant. There was no evidence of differential effectiveness by gender or SEP. This review provides the strongest evidence to date that current school-based efforts do not positively impact young people's physical activity across the full day, with no difference in effect across gender and SEP. Further assessment and maximization of implementation fidelity is required before it can be concluded that these interventions have no contribution to make.


Inhibition of intramacrophage growth of Penicillium marneffei by 4-aminoquinolines.
The antimicrobial activities of chloroquine (CQ) and several 4-aminoquinoline drugs were tested against Penicillium marneffei, an opportunistic fungus that invades and grows inside macrophages and causes disseminated infection in AIDS patients. Human THP1 and mouse J774 macrophages were infected in vitro with P. marneffei conidia and treated with different doses of drugs for 24 to 48 h followed by cell lysis and the counting of P. marneffei CFU. CQ and amodiaquine exerted a dose-dependent inhibition of fungal growth, whereas quinine and artemisinin were fungistatic and not fungicidal. The antifungal activity of CQ was not due to an impairment of fungal iron acquisition in that it was not reversed by the addition of iron nitrilotriacetate, FeCl3, or iron ammonium citrate. Perl's staining indicated that CQ did not alter the ability of J774 cells to acquire iron from the medium. Most likely, CQ's antifungal activity is due to an increase in the intravacuolar pH and a disruption of pH-dependent metabolic processes. Indeed, we demonstrate that (i) bafilomycin A1 and ammonium chloride, two agents known to alkalinize intracellular vesicles by different mechanisms, were inhibitory as well and (ii) a newly synthesized 4-amino-7-chloroquinoline molecule (compound 9), lacking the terminal amino side chain of CQ that assists in drug accumulation, did not inhibit P. marneffei growth. These results suggest that CQ has a potential for use in prophylaxis of P. marneffei infections in human immunodeficiency virus-infected patients in countries where P. marneffei is endemic.


Upregulation of the transport system for TNFalpha at the blood-brain barrier.
The transport system for the cytokine tumor necrosis factor-alpha (TNFalpha) at the blood-brain barrier (BBB) enables an enhanced yet saturable entry of TNFalpha from blood to the CNS. This review focuses on the selective upregulation of the transport system for TNFalpha at the BBB that is specific for type of pathology, region, and time. The upregulation is reflected by increased CNS tissue uptake of radiolabeled TNFalpha after iv injection in mice and by inhibition of this increase with excess non-radiolabeled TNFalpha. (1) Spinal cord injury (SCI): upregulation of TNFalpha uptake after thoracic transection is seen in the delayed phase of BBB disruption at the lumbar spinal cord. Thoracic SCI by compression, however, has a longer lasting impact on TNFalpha transport that involves thoracic and lumbar spinal cord, in contrast to the upregulation confined to the lumbar region in lumbar SCI by compression. Regardless, the uptake of TNFalpha by spinal cord does not parallel BBB disruption as measured by the leakage of radiolabeled albumin. (2) Experimental autoimmune encephalomyelitis (EAE): the increase in the differential permeability to TNFalpha is seen in all CNS regions (brain and cervical, thoracic, and lumbar spinal cord) and has a distinct time course and reversibility. Exogenous TNFalpha has biphasic effects in modulating functional scores. The BBB, a dynamically regulated barrier, is actively involved in disease processes.


[Vertebral artery loop and cervical pain].
Diagnosis of a vertebral artery loop in the transverse canal, suggested by a CT scan image, is dependent mainly on results of angiography (preferably computer assisted and by the venous route). This lesion is a differential diagnosis of a neuroma, when the cervical intervertebral foramen is enlarged.


Avoidance of medial cortical fracture in high tibial osteotomy: improved technique.
A new technique in oblique incomplete high tibial osteotomy that permits an increase of valgus correction while preventing fracture of the medial cortex was investigated. Closing wedge or opening wedge osteotomy was done on 23 tibias from cadavers before loading in an Instron testing machine. In seven specimens (Group 1), lateral oblique wedge osteotomy was done. In seven other specimens (Group 2), one medial oblique cut was made. In both groups, the osteotomy terminated 10 mm from the cortex and approximately 2 cm below the plateau. In nine specimens (Group 3), the osteotomy terminated in a 5-mm diameter hole, drilled in an anteroposterior direction, with its center positioned 10 mm from the medial cortex and 2 cm below the articular surface. The maximum angle of opening or closing before fracture of the cortex took place was recorded. In Groups 1 and 2, similar maximum correction angles were observed, 6.7 degrees versus 6.5 degrees, respectively. In Group 3, the stress relieving hole allowed the correction angle to be increased to 10 degrees. An oblique high tibial valgus closing wedge osteotomy with an apical drill hole allows a significant increase of the correction angle compared with the same osteotomy without a drill hole. Medial open wedge osteotomy offers no advantage over lateral closed wedge osteotomy in the maximum obtainable correction angle without failure of the cortex.


Effect of human alpha-antitrypsin in papain-induced emphysema in the hamster.
The possibility that human alpha1-antitrypsin could effectively prevent development of emphysematous lesions produced in hamsters 7 days after exposure to aerosolized papain (3% for 3 hours) was investigated. Pretreatment with intratracheal human alpha1-antitrypsin prevented the appearance of these lesions in a dose-dependent manner. On the other hand, systemic administration of large doses of human alpha1-antitrypsin failed to prevent papain-induced pulmonary lesions, despite a significant increase in serum trypsin inhibitory capacity. These results suggest that intra-alveolar rather than serum concentrations of human alpha1-antitrypsin are critical for preventing the development of pulmonary emphysema in this animal model. It is interesting that although the mechanism by which human alpha1-antitrypsin prevented the papain-induced lesions is unknown, intratracheally administered human alpha1-antitrypsin similarly prevented the development of pulmonary emphysematous lesions induced by 0.1 mg of porcine pancreatic elastase given intratracheally.


Evolutionary processes involved in the diversification of chelonian and mammal polystomatid parasites (Platyhelminthes, Monogenea, Polystomatidae) revealed by palaeoecology of their hosts.
Polystomatid flatworms (Platyhelminthes) are monogenean parasites that infect exclusively aquatic or semi-aquatic sarcopterygians such as the Australian lungfish, amphibians, freshwater turtles and the African common hippopotamus. Previous studies on the phylogenetic relationships of these parasites, excluding Oculotrema hippopotami infecting common hippos, showed a global coevolution between hosts and their parasites at a macroevolutionary scale. These studies also demonstrated a strong correlation between the diversification of early neobatrachian polystomes and Gondwana breakup in the Mesozoic period. However the origin of chelonian polystomes is still in question as a switch from presumably primitive aquatic amniotes to turtles at the time of their first appearance, or soon after during their radiation, was assumed. In order to resolve this sticking point, we extended the phylogeny of polystomes with broader parasite sampling, i.e. 55 polystome species including Nanopolystoma tinsleyi a polystome infecting caecilians and O. hippopotami, and larger set of sequence data covering two nuclear and two mitochondrial genes coding for the ribosomal RNA 18S and 28S, the Cytochrome c Oxidase I and the ribosomal RNA 12S, respectively. The secondary structure of nuclear rRNAs genes (stems and loops) was taken into account for sequence alignments and Bayesian analyses were performed based on the appropriate models of evolution selected independently for the four designed partitions. Molecular calibrations were also conducted for dating the main speciation events in the polystome tree. The phylogenetic position of chelonian parasites that are phylogenetically closer to N. tinsleyi than all other amphibian polystomes and molecular time estimates suggest that these parasites originated following a switch from caecilians, at a geological period when primitive turtles may already have adapted to an aquatic life style, i.e. at about 178Million years ago, or a little later when the crown group of extant turtles have already diversified, i.e. at about 152Mya. Similarly, because O. hippopotami constitutes the sister group of chelonian parasites, proposing that an African caecilian could be the ancestral host for this polystome species seems at this stage the most likely hypothesis to explain its occurrence within the common hippo. Regardless of the scenario that may be predicted to explain the origin of polystomes within aquatic or semi-aquatic amniotes, their presence and evolution are indicative of early aquatic ecological habits within ancestral lineages.


4-Acetyl-3-[2-(eth-oxy-carbon-yl)phen-yl]sydnone.
Sydnones, which contain a mesoionic five-membered heterocyclic ring, are more stable if synthesized with an aromatic substutuent at the N3 position. In the title compound {sys-tematic name: 4-acetyl-3-[2-(eth-oxy-carbon-yl)phen-yl]-1,2,3-oxa-diazol-3-ylium-5-olate}, C13H12N2O5, the aromatic substitutent is 2-(eth-oxy-carbon-yl)phenyl. Intra- and inter-molecular hydrogen bonds are observed. The inter-planar angle between the sydnone and benzene rings is 71.94 (8)°. π-ring⋯carbon-yl inter-actions of 3.2038 (16) Å arise between the sydnone ring and a symmetry-related C=O group.


Pushing the limit: investigation of hydrodynamic forces on a trapped particle kicked by a laser pulse.
We introduce a new optical technique where a train of short optical pulses is utilized to disturb a trapped microscopic particle. Using fast (250 kHz) and accurate (nm) detection of the position of the particle, accurately synchronized to the repetition rate of the laser pulses, we can coherently superimpose the displacement caused by each individual laser pulse. Thereby we are able to both bypass the influence from the Brownian motion of the trapped particle and to simultaneously increase the ability to localize its average trajectory by √n, where n is the number of repetitive pulses. In the results presented here we utilize a train of 1200 pulses to kick a 5 μm polystyrene sphere and obtain a spatial resolution corresponding to 0.09 nm and a time resolution of 4 μs. The magnitude of the optical force pushing the particle corresponds to ∼ 10(4)g and enables an investigation of both the hydrodynamical drag and the inertial effects caused by the particle and the surrounding liquid. Our results enables a more accurate testing of the existing extended models for the hydrodynamic drag and we discuss the observed agreement between experiments and theory.


The discrepancy between 99Tcm-ECD dynamic and static SPET images in patients with ischaemic lesions corresponds to reduced vasoreactivity to acetazolamide.
We evaluated 99Tcm-N,N'-(1,2-ethylenediyl)bis-L-cysteine diethyl ester (99Tcm-ECD) dynamic and static SPET (single photon emission tomographic) images to examine 99Tcm-ECD kinetics under ischaemic cerebrovascular conditions. In 20 patients who showed arterial occlusion on magnetic resonance angiography, dynamic (0-10 min) and static (15-35 min) SPET images were acquired after the intravenous administration of 99Tcm-ECD. Thirteen of the patients had focal perfusion deficits that were more evident on the dynamic than on the static images; the other seven showed no such discrepancy. In those patients with a mismatch between the dynamic and static images, the extent corresponded to reduced vaso-reactivity to acetazolamide. Based on quantitative analysis of the ratio of tracer uptake in affected to that in unaffected areas, the patients with discrepant findings showed significantly different ratios on the dynamic and static images, whereas those with no such mismatch did not. Our results suggest that dynamic 99Tcm-ECD images provide circulatory information and that static images reflect a filling-in phenomenon of ECD metabolites in ischaemic lesions. 99Tcm-ECD dynamic and static SPET images offer an alternative method of detecting mild perfusion deficits without the need for acetazolamide challenge.


Development of a new prognostic index PNPI for prognosis prediction of CKD patients with pneumonia at hospital admission.
The aim of this study was to investigate the relationship between pneumonia and chronic kidney disease (CKD), to elucidate potential risk factors, and to develop a new predictive model for the poor prognosis of pneumonia in CKD patients. We conducted a retrospective observational study of CKD patients admitted to Tongji Hospital between June 2012 and June 2022. Demographic information, comorbidities or laboratory tests were collected. Applying univariate and multivariate logistic regression analyses, independent risk factors associated with a poor prognosis (i.e., respiratory failure, shock, combined other organ failure, and/or death during hospitalization) for pneumonia in CKD patients were discovered, with nomogram model subsequently developed. Predictive model was compared with other commonly used pneumonia severity scores. Of 3,193 CKD patients with pneumonia, 1,013 (31.7%) met the primary endpoint during hospitalization. Risk factors predicting poor prognosis of pneumonia in CKD patients were selected on the result of multivariate logistic regression models, including chronic cardiac disease; CKD stage; elevated neutrophil to lymphocyte ratio (NLR) and D-dimer; decreased platelets, PTA, and chloride iron; and significant symptom presence and GGO presentation on CT. The nomogram model outperformed other pneumonia severity indices with AUC of 0.82 (95% CI: 0.80, 0.84) in training set and 0.83 (95% CI: 0.80, 0.86) in testing set. In addition, calibration curve and decision curve analysis (DCA) proved its efficiency and adaptability. We designed a clinical prediction model PNPI (pneumonia in nephropathy patients prognostic index) to assess the risk of poor prognosis in CKD patients with pneumonia, which may be generalized after more external validation.


Development of the gas puff charge exchange recombination spectroscopy (GP-CXRS) technique for ion measurements in the plasma edge.
A novel charge-exchange recombination spectroscopy (CXRS) diagnostic method is presented, which uses a simple thermal gas puff for its donor neutral source, instead of the typical high-energy neutral beam. This diagnostic, named gas puff CXRS (GP-CXRS), is used to measure ion density, velocity, and temperature in the tokamak edge/pedestal region with excellent signal-background ratios, and has a number of advantages to conventional beam-based CXRS systems. Here we develop the physics basis for GP-CXRS, including the neutral transport, the charge-exchange process at low energies, and effects of energy-dependent rate coefficients on the measurements. The GP-CXRS hardware setup is described on two separate tokamaks, Alcator C-Mod and ASDEX Upgrade. Measured spectra and profiles are also presented. Profile comparisons of GP-CXRS and a beam based CXRS system show good agreement. Emphasis is given throughout to describing guiding principles for users interested in applying the GP-CXRS diagnostic technique.


Aging, frailty and age-related diseases.
The concept of frailty as a medically distinct syndrome has evolved based on the clinical experience of geriatricians and is clinically well recognizable. Frailty is a nonspecific state of vulnerability, which reflects multisystem physiological change. These changes underlying frailty do not always achieve disease status, so some people, usually very elderly, are frail without a specific life threatening illness. Current thinking is that not only physical but also psychological, cognitive and social factors contribute to this syndrome and need to be taken into account in its definition and treatment. Together, these signs and symptoms seem to reflect a reduced functional reserve and consequent decrease in adaptation (resilience) to any sort of stressor and perhaps even in the absence of extrinsic stressors. The overall consequence is that frail elderly are at higher risk for accelerated physical and cognitive decline, disability and death. All these characteristics associated with frailty can easily be applied to the definition and characterization of the aging process per se and there is little consensus in the literature concerning the physiological/biological pathways associated with or determining frailty. It is probably true to say that a consensus view would implicate heightened chronic systemic inflammation as a major contributor to frailty. This review will focus on the relationship between aging, frailty and age-related diseases, and will highlight possible interventions to reduce the occurrence and effects of frailty in elderly people.


[So-called endometrial carcinoma of the male prostatic utricle].
From a review of the literature the author concludes that the histogenesis of the so-called endometrial carcinoma of the male prostatic utricle first described by Melicow and Pachter in 1967 is not completely clear yet. It may be assumed that tumors of a similar histological structure may be of different histogenesis, utricular and ductal, and that there are fine structural, histochemical, and histoenzymological differences between them which have not yet been studied sufficiently. It is suggested that the use of the immunoperoxidase method may be promising in such studies.


The first century of the "clinica del lavoro" in Milan.
The Clinica del Lavoro was created in Milan at the beginning of the 20th century by Luigi Devotto, who deemed it essential for physicians and health researchers to get involved in the life and health problems of working populations. The main roles of the Clinica del Lavoro were to educate medical students and train physicians; study actual workplaces, examine health and safety hazards and their noxious effects; and create initiatives and services to protect and promote workers' health. Important scientific contributions were made in several fields, including chemical carcinogenesis, effects of mineral and biologicalfibers, mechanism of action of silica dust, methods for the detection and measurements of toxic substances in both the work environment and workers' biological media.


The effectiveness of cardiology consultation. Concordance with diagnostic and drug recommendations.
In an effort to characterize the effectiveness of cardiology consultation, the outcomes of consultants' recommendations for diagnostic actions and cardiac drugs were quantitatively examined in 394 cases. Drug recommendations were made more frequently (49 percent) than diagnostic recommendations (38 percent) and were associated with a higher rate of concordance (82 percent to 64 percent). Recommendations to start therapy with a drug, especially an antihypertensive or an antianginal, were associated with a lower concordance rate than recommendations to continue or discontinue drug therapy. Consultees' responses to recommendations for diagnostic action did not vary significantly according to the type of action suggested. Nonconcordance with diagnostic suggestions was particularly high when a service made a large number of consult requests. Concordance was increased if the consultant left a follow-up note. Consultees' responses to drug and diagnostic recommendations were independent of one another. The study represents the first systematic assessment of cardiology consultation activities and provides a methodology for subsequent studies.


Localization of neural cell adhesion molecule and pan-cadherin immunoreactivity in intrauterine growth-retarded newborn rat kidneys.
To examine the expression and distribution pattern of neural cell adhesion molecule (N-CAM) and pan-cadherin immunoreactivity in intrauterine growth retardation (IUGR) newborn kidneys, we used a rat model of maternal protein restriction throughout pregnancy. Weak or moderate immunoreactivity for N-CAM and pan-cadherin was seen in some proximal tubules, in the thick segments of Henle, and in the collecting tubules of the control sections. However, the number of tubules expressing N-CAM and pan-cadherin was increased in the IUGR group compared with the control group. Increased density of N-CAM and pan-cadherin immunoreactivity was observed mostly in the proximal tubules, in the thick segments of Henle, and in the collecting tubules of IUGR newborn rat kidneys. Furthermore, N-CAM and pan-cadherin immunoreactivity was present in the thin limb of Henle in the IUGR group, whereas it was absent in the control group. Glomeruli were negative in both groups except for some glomeruli that showed very weak N-CAM staining in the IUGR group. Thus it was demonstrated for the first time that IUGR newborn rat kidneys express N-CAM and cadherin adhesion molecules at specific sites of the nephron.


An Online, Person-Centered, Risk Factor Management Program to Prevent Cognitive Decline: Protocol for A Prospective Behavior-Modification Blinded Endpoint Randomized Controlled Trial.
Several modifiable risk factors for dementia have been identified, although the extent to which their modification leads to improved cognitive outcomes remains unclear. The primary aim is to test the hypothesis that a behavior modification intervention program targeting personalized risk factors prevents cognitive decline in community-dwelling, middle-aged adults with a family history of dementia. This is a prospective, risk factor management, blinded endpoint, randomized, controlled trial, where 1510 cognitively normal, community-dwelling adults aged 40-70 years old will be recruited. Participants will be screened for risk factors related to vascular health (including physical inactivity), mental health, sleep, and cognitive/social engagement. The intervention is an online person-centered risk factor management program: BetterBrains. Participants randomized to intervention will receive telehealth-based person-centered goal setting, motivational interviewing, and follow-up support, health care provider communication and community linkage for management of known modifiable risk factors of dementia. Psychoeducational health information will be provided to both control and intervention groups. The primary outcome is favorable cognitive performance at 24-months post-baseline, defined as the absence of decline on one or more of the following cognitive tests: (a) Cogstate Detection, (b) Cogstate One Card Learning, (c) Cogstate One Back, and (d) Cognitive Function Instrument total score. We will test the hypothesis that the BetterBrains intervention program can prevent cognitive decline. By leveraging existing community services and using a risk factor management pathway that tailors the intervention to each participant, we maximize likelihood for engagement, long-term adherence, and for preserving cognitive function in at-risk individuals.


[Occupation-, transportation- and leisure-related physical activity: gender inequalities in Santander, Colombia].
Objectives To estimate the prevalence of occupation-, transportation- and leisure-related physical activity, its compliance with recommendations, and to explore its association with demographic and socioeconomic variables in men and women of the Department of Santander (Colombia). Methods The sample consisted of 2421 people between 15 and 64 years of age, participants in the Risk Factors for Chronic Diseases of Santander cross-sectional study, developed in 2010. The Global Physical Activity Questionnaire was used for data collection. Age-adjusted prevalence ratios were calculated and multivariate analysis models were built by sex using robust Poisson regression. Results The prevalence of occupational and leisure physical activity and compliance with recommendations were lower in women. Sexual division of labor and a low socioeconomic level negatively influenced physical activity in women, limiting the possibility of practice of those principally engaged in unpaid work at home. Young or single men and those living in higher socioeconomic areas were more likely to practice physical activity in leisure time and meet recommendations. Conclusion Physical activity surveillance and related public policies should take into account the inequalities between the practice of men and women related to their socioeconomic conditions and the sexual division of labor.


Industrial Research as a "Corporate Counterculture"? The Development of the HPO Caprolactam Process at DSM, 1956-77.
This paper focuses on the development of the hydroxylamine phosphate oxime (HPO) process for the manufacture of caprolactam, an intermediate for nylon, by Central Laboratory, the R&D organisation of the Dutch chemical company DSM. The example of the HPO process shows that Central Laboratory was independent rather than isolated. This position enabled it to develop its own view of the interest of the company and start research projects regardless of the opinion of the production and marketing functions. The example of the HPO process shows the potency of such a position but also the problems involved. As a medium-sized chemical company, DSM is a company that may be more typical for the practice of R&D between 1945 and 1970 than the large companies that historians have typically chosen for analysis.


Biomaterials in orthopedic surgery: effects of a nickel-reduced stainless steel on in vitro proliferation and activation of human osteoblasts.
A new austenitic stainless steel compound, P558, has been widely recognized to have good mechanical properties, excellent potential for corrosion resistance and negligible nickel ion release, making it a promising substitute for more expensive metallic prostheses with limited machinable features. The effect of P558 was studied in vitro and human osteoblast- like cells (MG63) were cultured directly on P558, Ti6Al4V alloy (Ti), and polystyrene (Control) for 72 hours. Osteoblast functions were evaluated by assaying cell proliferation and synthetic activity after 1.25(OH)2D3 stimulation. Results demonstrated that growth of MG63 on P558 was not negatively affected when compared to the Ti and Control groups and showed no alteration in the production of ALP, NO and PICP. Moreover, IL-6 was lower, whereas OC and TGFbeta1 were significantly higher. SEM images revealed that cells proliferated and differentiated on P558 without any alteration in their morphology. The current findings have demonstrated that P558 promotes osteoblast proliferation, activation and differentiation without negative effects and, thus, its good biocompatibility when used for orthopedic application.


Mathematical models for the spatial receptive-field organization of nonlagged X-cells in dorsal lateral geniculate nucleus of cat.
Spatial receptive fields of relay cells in dorsal lateral geniculate nucleus (dLGN) have commonly been modeled as a difference of two Gaussian functions. We present alternative models for dLGN cells which take known physiological couplings between retina and dLGN and within dLGN into account. The models include excitatory input from a single retinal ganglion cell and feedforward inhibition via intrageniculate interneurons. Mathematical formulas describing the receptive field and response to circular spot stimuli are found both for models with a finite and an infinite number of ganglion-cell inputs to dLGN neurons. The advantage of these models compared to the common difference-of-Gaussians model is that they, in addition to providing mathematical descriptions of the receptive fields of dLGN neurons, also make explicit contributions from the geniculate circuit. Moreover, the model parameters have direct physiological relevance and can be manipulated and measured experimentally. The discrete model is applied to recently published data (Ruksenas et al., 2000) on response versus spot-diameter curves for dLGN cells and for the retinal input to the cell (S-potentials). The models are found to account well for the results for the X-cells in these experiments. Moreover, predictions from the discrete model regarding receptive-field sizes of interneurons, the amount of center-surround antagonism for interneurons compared to relay cells, and distance between neighboring retinal ganglion cells providing input to interneurons, are all compatible with data available in the literature.


[Study of the effectiveness of ampicillin in the therapy of fresh gonorrhea in men and women].
The results of ampicillin use in therapy of 114 cases with fresh gonorrhea are presented. Males with acute and subacute urethritis and females with gonorrhea of the lower part of the genitourinary system were treated with ampicillin in the total dose of 2.5 gm according to the schedule: 1--0.5--0.5--0.5 gm every 6 hours. For the treatment of cases with torpid, prostatitis complicated, ascending gonorrhea combined with trichomonasis and in patients weighing more than 70 kg the course dose was increased up to 3.5--6 gm. Positive results were observed in 99.1 per cent of the cases. Relapses of the disease were registered in 0.9 per cent of the patients. Postgonorrhea changes were found in 16.7 per cent (7.5 per cent of the males and 38.1 per cent of the females). A definite conformity between sensitivity of gonococci to benzylpenicillin and ampicillin was shown, which should be taken into account in cases with increased resistance of gonococci to one of the drugs. Ampicillin is an effective antigonorrhea drug. Clearance of the focal exudates from gonococci was observed during the first 24 hours of the treatment.


Development of an ex vivo model for the study of cerebrovascular function utilizing isolated mouse olfactory artery.
Cerebral vessels, such as intracerebral perforating arterioles isolated from rat brain, have been widely used as an ex vivo model to study the cerebrovascular function associated with cerebrovascular disorders and the therapeutic effects of various pharmacological agents. These perforating arterioles, however, have demonstrated differences in the vascular architecture and reactivity compared with a larger leptomeningeal artery which has been commonly implicated in cerebrovascular disease. In this study, therefore, we developed the method for studying cerebrovascular function utilizing the olfactory artery isolated from the mouse brain. The olfactory artery (OA) was isolated from the C57/BL6 wild-type mouse brain. After removing connective tissues, one side of the isolated vessel segment (approximately -500 µm in length) was cannulated and the opposite end of the vessel was completely sealed while being viewed with an inverted microscope. After verifying the absence of pressure leakage, we examined the vascular reactivity to various vasoactive agents under the fixed intravascular pressure (60 mm Hg). We found that the isolated mouse OAs were able to constrict in response to vasoconstrictors, including KCl, phenylephrine, endothelin-1, and prostaglandin PGH2. Moreover, this isolated vessel demonstrated vasodilation in a dose-dependent manner when vasodilatory agents, acetylcholine and bradykinin, were applied. Our findings suggest that the isolated olfactory artery would provide as a useful ex vivo model to study the molecular and cellular mechanisms of vascular function underlying cerebrovascular disorders and the direct effects of such disease-modifying pathways on cerebrovascular function utilizing pharmacological agents and genetically modified mouse models.


Quantitative variation of pheromone components in the spruce bark beetleIps typographus from different attack phases.
Ips typographus beetles were collected in the field, separated into eight attack phases (from beetles walking on the trunk of a tree under attack to those excavating gallery systems with a mother gallery longer than 4 cm), and immediately frozen in liquid nitrogen. 2-Methyl-3-buten-2-ol,cis- andtrans-verbenol, verbenone, myrtenol, trans-myrtanol, ipsenol, ipsdienol, and 2-phenylethanol were quantified from excised hindguts against an internal standard, heptyl acetate, in the extraction solvent. Methylbutenol, the pinene alcohols, and 2-phenylethanol showed the same pattern of variation between attack phases in males, with the largest amounts present before accepting females and then a fast decline. Ipsenol and ipsdienol were not detected in males before the females were accepted, and the amounts increased when the females start their egg laying. Verbenone occurred only in trace amounts. The beetles were sampled from five Norway spruce trees (Picea abies) of differing resin flow. The correlations between the nine pheromone components and five major host monoterpenes in the gut showed that the variation in the amount of methyl-butenol, ipsenol, and ipsdienol could not be explained by the variation in the amounts of host monoterpenes. In contrast over 80% of the quantitative variation ofcis-verbenol,trans-verbenol, and myrtenol was explained by the amount of α-pinene. The nine pheromone components from 36 individual males were also quantified. Both methylbutenol andcis-verbenol showed a large variation in both amounts and proportions. Females containedtrans-verbenol and traces of most other components found in males. When accepted by the male, they also contained a female-specific compound, β-isophorone. Behavioral and biosynthetic implications of the results are discussed.


MANAGEMENT OF ENDOCRINE DISEASE: Does gender matter in the management of acromegaly?
Gonadal steroids modulate the effects of GH, with oestrogens attenuating and androgens augmenting GH action. Whether these divergent effects influence the clinical manifestation, management and prognosis of acromegaly have not been carefully reviewed. This review examines whether there is a gender difference in epidemiology, presentation, quality of life (QoL), morbidity, treatments and mortality of acromegaly. Acromegaly is more common in women who present at an older age with longer diagnostic delay. At presentation, women have a higher GH relative to IGF-1 level than men. QoL is more adversely affected in women both before and after treatment. Prevalence of hypertension and diabetes are greater in women than in men with acromegaly. Treatment outcomes with SSAs are comparable between sexes, but women may require a higher dose of pegvisomant for equivalent response. Mortality in untreated acromegaly is more profoundly affected in women; however, improved treatments in recent decades have resulted in normalisation of standard mortality ratios in both sexes. We conclude that gender does matter in the management of acromegaly, with women presenting later in life, with greater diagnostic delay, higher prevalence of comorbidities and experiencing worse QoL.


Phylogenetic evidence that the gram-negative nonsporulating bacterium Tissierella (Bacteroides) praeacuta is a member of the Clostridium subphylum of the gram-positive bacteria and description of Tissierella creatinini sp. nov.
The 16S rRNA gene sequence of the type strain of Tissierella praeacuta (formerly Bacteroides praeacutus) was determined by PCR direct sequencing. A comparative sequence analysis showed that T. praeacuta is a member of the Clostridium subphylum of the gram-positive bacteria and has a close phylogenetic affinity with the species that form Clostridium cluster XII (M. D. Collins, P. A. Lawson, A. Willems, J. J. Cordoba, J. Fernandez-Garayzabal, P. Garcia, J. Cai, H. Hippe, and J. A. E. Farrow, Int. J. Syst. Bacteriol. 44:812-826, 1994). Although T. praeacuta is gram negative and does not produce endospores, 16S rRNA sequence data showed that it is closely related genealogically (level of sequence similarity, 99.9%) to Clostridium hastiforme. On the basis of our results and the results of previous studies, a second species of Tissierella, Tissierella creatinini sp. nov., is described.


Activity of the Mason-Pfizer monkey virus fusion protein is modulated by single amino acids in the cytoplasmic tail.
Mason-Pfizer monkey virus (M-PMV) encodes a transmembrane glycoprotein with a 38-amino-acid-long cytoplasmic tail. After the release of the immature virus, a viral protease-mediated cleavage of the cytoplasmic tail (CT) results in the loss of 17 amino acids from the carboxy terminus and renders the envelope protein fusion competent. To investigate the role of individual amino acid residues in the CT in fusion, a series of mutations was introduced, and the effects of these mutations on glycoprotein biosynthesis and fusion were examined. Most of the alanine-scanning mutations in the CT had little effect on fusion activity. However, four amino acid substitutions (threonine 4, lysine 7, glutamine 9, and isoleucine 10) resulted in substantially increased fusogenicity, while six (leucine 2, phenylalanine 5, isoleucine 13, lysine 16, proline 17, and glycine 31) resulted in much-reduced fusion. Interestingly, the bulk of these mutations are located upstream of the CT cleavage site in a region that has the potential to form a coiled-coil in the Env trimer. Substitutions at glutamine 9 and isoleucine 10 with alanine had the most dramatic positive effect and resulted in the formation of large syncytia. Taken together, these data demonstrate that individual residues within the cytoplasmic domain of M-PMV Env can modulate, in both a positive and negative manner, biological functions that are associated with the extracellular domains of the glycoprotein complex.


Th17-related cytokines: new players in the control of chronic intestinal inflammation.
Crohn's disease (CD) and ulcerative colitis (UC), the main forms of inflammatory bowel diseases (IBD) in man, are thought to be caused by an excessive and poorly controlled immune response that is directed against components of the normal microflora. The exact sequence of events by which this pathological process is triggered and maintained is not fully understood, but studies in experimental models of IBD and data emerging from recent clinical trials indicate that T cell-derived cytokines are crucial mediators of the tissue damage. Although CD and UC have been traditionally considered two typical examples of T helper (Th)1 or Th2-associated disease respectively, it is now known that CD- and UC-related inflammation is also marked by enhanced production of cytokines made by a distinct subset of Th cells, termed Th17 cells. Th17 cytokines can have both tissue-protective and inflammatory effects in the gut and there is evidence that Th17 cells can alter their cytokine program according to the stimuli received and convert into Th1-producing cells. These novel findings have contributed to advancing our understanding of mechanisms of gut tissue damage and open new avenues for development of therapeutic strategies in IBD.


Autophagy Accompanied with Bisdemethoxycurcumin-induced Apoptosis in Non-small Cell Lung Cancer Cells.
To investigate the effects of bisdemethoxycurcumin (BDMC) on non-small cell lung cancer (NSCLC) cell line, A549, and the highly metastatic lung cancer 95D cells. CCK-8 assay was used to assess the effect of BDMC on cytotoxicity. Flow cytometry was used to evaluate apoptosis. Western blot analysis, electron microscopy, and quantification of GFP-LC3 punctuates were used to test the effect of BDMC on autophagy and apoptosis of lung cancer cells. BDMC inhibited the viability of NSCLC cells, but had no cytotoxic effects on lung small airway epithelial cells (SAECs). The apoptotic cell death induced by BDMC was accompanied with the induction of autophagy in NSCLC cells. Blockage of autophagy by the autophagy inhibitor 3-methyladenine (3-MA) repressed the growth inhibitory effects and induction of apoptosis by BDMC. In addition, BDMC treatment significantly decreased smoothened (SMO) and the transcription factor glioma-associated oncogene 1 (Gli1) expression. Furthermore, depletion of Gli1 by siRNA and cyclopamine (a specific SMO inhibitor) induced autophagy. Aberrant activation of Hedgehog (Hh) signaling has been implicated in several human cancers, including lung cancers. The present findings provide direct evidence that BDMC-induced autophagy plays a pro-death role in NSCLC, in part, by inhibiting Hedgehog signaling.


Pathologists as stewards of laboratory information.
Just as electronic health records are transforming the practice of medicine and health care information management, practicing in the era of the electronic health record offers opportunities, if not imperatives, for pathologists to take on new and "transformative" professional and leadership roles for the organizations they serve. Experience indicates that clinicians will perceive pathologists and laboratories as responsible for all aspects of laboratory testing and information management, including order entry and results reporting, even though such functions may fall beyond the control of the laboratory. As described and expanded upon in the previous 4 articles of this series, the use of electronic health records dictates changes in how clinicians interact with laboratory information. In this environment, pathologists are uniquely positioned to act as the stewards for laboratory information in electronic health records and throughout health care organizations.


Thermoconformational Behavior of Cellulose Nanofiber Films as a Device Substrate and Their Superior Flexibility and Durability to Glass.
The design and high-throughput manufacturing of thin renewable energy devices with high structural and atomic configurational stability are crucial for the fabrication of green electronics. Yet, this concept is still in its infancy. In this work, we report the extraordinary durability of thin molecular interlayered organic flexible energy devices based on chemically tuned cellulose nanofiber transparent films that outperform glass by decreasing the substrate weight by 50%. The nanofabricated flexible thin film has an exceptionally low thermal coefficient of expansion of 1.8 ppm/K and a stable atomic configuration under a harsh fabrication condition (over 190 °C for an extended period of 5 h). A flexible optoelectronic device using the same renewable cellulose nanofiber film substrate was found to be functionally operational over a life span of 5 years under an intermittent operating condition. The success of this device's stability opens up an entirely new frontier of applications of flexible electronics.


Paradoxical enhancement of letter recognition in developmental dyslexia.
In a number of studies, children diagnosed with developmental dyslexia reached normal scores in standard visual processing tasks, and some researchers concluded that visual processing deficits are not involved in the syndrome. The tasks used, however, may be insensitive to anomalous visual information processing strategies used to compensate for an underlying deficit. To determine whether children with dyslexia use anomalous visual processing strategies, a same-different task was applied, in which 2 items identical under rotation and reflection were judged as same. Pairs of letters or dot patterns were used, which were either symmetric or asymmetric in shape. Children with dyslexia performed faster than normal-reading children--in particular, remarkably, with letters. Symmetry of dot patterns facilitated performance in both children with dyslexia and normal-reading children; symmetry of letters facilitated performance in children with dyslexia but not in normal-reading children. Children with dyslexia, therefore, fail to adequately differentiate visual processing of linguistic and non-linguistic materials; they process symmetry in letters similarly to that in shapes, which leads in this particular task to the paradoxical observation of children with dyslexia outperforming normal readers with letters.


The state of health services in China and India: a larger context.
In this paper the problems of health services in China and India are related to some structural features of the two economies. Some similarities and differences exist across these two countries in terms of political economy, with differential results. Both countries have experienced remarkable economic growth during the past quarter-century, but this has not always translated into improvements in health for the poor. Although China used to have an egalitarian basic public health service, the system has become quite inegalitarian during the past quarter-century, with the disintegration of the communes and adoption of fee-based services under a system of decentralized public finance. India's health system has remained inegalitarian throughout.


Degradation process of lead chromate in paintings by Vincent van Gogh studied by means of spectromicroscopic methods. Part 5. Effects of nonoriginal surface coatings into the nature and distribution of chromium and sulfur species in chrome yellow paints.
The darkening of lead chromate yellow pigments, caused by a reduction of the chromate ions to Cr(III) compounds, is known to affect the appearance of several paintings by Vincent van Gogh. In previous papers of this series, we demonstrated that the darkening is activated by light and depends on the chemical composition and crystalline structure of the pigments. In this work, the results of Part 2 are extended and complemented with a new study aimed at deepening the knowledge of the nature and distribution of Cr and S species at the interface between the chrome yellow paint and the nonoriginal coating layer. For this purpose, three microsamples from two varnished paintings by Van Gogh and a waxed low relief by Gauguin (all originally uncoated) have been examined. Because nonoriginal coatings are often present in artwork by Van Gogh and contemporaries, the understanding of whether or not their application has influenced the morphological and/or physicochemical properties of the chrome yellow paint underneath is relevant in view of the conservation of these masterpieces. In all the samples studied, microscopic X-ray fluorescence (μ-XRF) and X-ray absorption near edge structure (μ-XANES) investigations showed that Cr(III)-based alteration products are present in the form of grains inside the coating (generally enriched of S species) and also homogeneously widespread at the paint surface. The distribution of Cr(III) species may be explained by the mechanical friction caused by the coating application by brush that picked up and redistributed the superficial Cr compounds, likely already present in the reduced state as result of the photodegradation process. The analysis of the XANES profiles allowed us to obtain new insights into the nature of the Cr(III) alteration products, that were identified as sulfate-, oxide-, organo-metal-, and chloride-based compounds. Building upon the knowledge acquired through the examination of original paint samples and from the investigation of aged model paints in the last Part 4 paper, in this study we aim to characterize a possible relation between the chemical composition of the coating and the chrome yellow degradation pathways by studying photochemically aged model samples covered with a dammar varnish contaminated with sulfide and sulfate salts. Cr speciation results did not show any evidence of the active role of the varnish and added S species on the reduction process of chrome yellows.


Common bile duct obstruction secondary to infection with Candida.
A patient is reported who had biliary tract obstruction secondary to infection of the common bile duct with Candida albicans, with the formation of a fungus ball. Treatment consisted of surgical removal of the fungus ball and drainage. Chemotherapy was not necessary. Ureteral obstruction through fungus ball formation, and even pulmonary fungus ball formation, has been attributed to candida, but this is the first case reported, to our knowledge, of bile duct obstruction.


Mitochondrial biogenesis, switching the sorting pathway of the intermembrane space receptor Mia40.
Mitochondrial precursor proteins are directed into the intermembrane space via two different routes, the presequence pathway and the redox-dependent MIA pathway. The pathways were assumed to be independent and transport different proteins. We report that the intermembrane space receptor Mia40 can switch between both pathways. In fungi, Mia40 is synthesized as large protein with an N-terminal presequence, whereas in metazoans and plants, Mia40 consists only of the conserved C-terminal domain. Human MIA40 and the C-terminal domain of yeast Mia40 (termed Mia40(core)) rescued the viability of Mia40-deficient yeast independently of the presence of a presequence. Purified Mia40(core) was imported into mitochondria via the MIA pathway. With cells expressing both full-length Mia40 and Mia40(core), we demonstrate that yeast Mia40 contains dual targeting information, directing the large precursor onto the presequence pathway and the smaller Mia40(core) onto the MIA pathway, raising interesting implications for the evolution of mitochondrial protein sorting.


Defective Excitatory/Inhibitory Synaptic Balance and Increased Neuron Apoptosis in a Zebrafish Model of Dravet Syndrome.
Dravet syndrome is a type of severe childhood epilepsy that responds poorly to current anti-epileptic drugs. In recent years, zebrafish disease models with Scn1Lab sodium channel deficiency have been generated to seek novel anti-epileptic drug candidates, some of which are currently undergoing clinical trials. However, the spectrum of neuronal deficits observed following Scn1Lab depletion in zebrafish larvae has not yet been fully explored. To fill this gap and gain a better understanding of the mechanisms underlying neuron hyperexcitation in Scn1Lab-depleted larvae, we analyzed neuron activity in vivo using combined local field potential recording and transient calcium uptake imaging, studied the distribution of excitatory and inhibitory synapses and neurons as well as investigated neuron apoptosis. We found that Scn1Lab-depleted larvae displayed recurrent epileptiform seizure events, associating massive synchronous calcium uptakes and ictal-like local field potential bursts. Scn1Lab-depletion also caused a dramatic shift in the neuronal and synaptic balance toward excitation and increased neuronal death. Our results thus provide in vivo evidence suggesting that Scn1Lab loss of function causes neuron hyperexcitation as the result of disturbed synaptic balance and increased neuronal apoptosis.


Genomic Prediction Informed by Biological Processes Expands Our Understanding of the Genetic Architecture Underlying Free Amino Acid Traits in Dry Arabidopsis Seeds.
Plant growth, development, and nutritional quality depends upon amino acid homeostasis, especially in seeds. However, our understanding of the underlying genetics influencing amino acid content and composition remains limited, with only a few candidate genes and quantitative trait loci identified to date. Improved knowledge of the genetics and biological processes that determine amino acid levels will enable researchers to use this information for plant breeding and biological discovery. Toward this goal, we used genomic prediction to identify biological processes that are associated with, and therefore potentially influence, free amino acid (FAA) composition in seeds of the model plant Arabidopsis thaliana Markers were split into categories based on metabolic pathway annotations and fit using a genomic partitioning model to evaluate the influence of each pathway on heritability explained, model fit, and predictive ability. Selected pathways included processes known to influence FAA composition, albeit to an unknown degree, and spanned four categories: amino acid, core, specialized, and protein metabolism. Using this approach, we identified associations for pathways containing known variants for FAA traits, in addition to finding new trait-pathway associations. Markers related to amino acid metabolism, which are directly involved in FAA regulation, improved predictive ability for branched chain amino acids and histidine. The use of genomic partitioning also revealed patterns across biochemical families, in which serine-derived FAAs were associated with protein related annotations and aromatic FAAs were associated with specialized metabolic pathways. Taken together, these findings provide evidence that genomic partitioning is a viable strategy to uncover the relative contributions of biological processes to FAA traits in seeds, offering a promising framework to guide hypothesis testing and narrow the search space for candidate genes.


Analgesic effects of butorphanol and buprenorphine in conscious African grey parrots (Psittacus erithacus erithacus and Psittacus erithacus timneh).
To evaluate effects of butorphanol tartrate and buprenorphine hydrochloride on withdrawal threshold to a noxious stimulus in conscious African grey parrots. 29 African grey parrots (Psittacus erithacus erithacus and Psittacus erithacus timneh). Birds were fitted with an electrode on the medial metatarsal region of the right leg, placed into a test box, and allowed to acclimate. An electrical stimulus (range, 0.0 to 1.46 mA) was delivered to each bird's foot through an aluminum perch. A withdrawal response was recorded when the bird lifted its foot from the perch or vigorously flinched its wings. Baseline threshold to a noxious electrical stimulus was determined. Birds then were randomly assigned to receive an i.m. injection of saline (0.9% NaCl) solution, butorphanol (1.0 mg/kg of body weight), or buprenorphine (0.1 mg/kg), and threshold values were determined again. Butorphanol significantly increased threshold value, but saline solution or buprenorphine did not significantly change threshold values. Butorphanol had an analgesic effect, significantly increasing the threshold to electrical stimuli in African grey parrots. Buprenorphine at the dosage used did not change the threshold to electrical stimulus. Butorphanol provided an analgesic response in half of the birds tested. Butorphanol would be expected to provide analgesia to African grey parrots in a clinical setting.


Structural and functional analysis of two new positive allosteric modulators of GluA2 desensitization and deactivation.
At the dimer interface of the extracellular ligand-binding domain of α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptors a hydrophilic pocket is formed that is known to interact with two classes of positive allosteric modulators, represented by cyclothiazide and the ampakine 2H,3H,6aH-pyrrolidino(2,1-3',2')1,3-oxazino(6',5'-5,4)benzo(e)1,4-dioxan-10-one (CX614). Here, we present structural and functional data on two new positive allosteric modulators of AMPA receptors, phenyl-1,4-bis-alkylsulfonamide (CMPDA) and phenyl-1,4-bis-carboxythiophene (CMPDB). Crystallographic data show that these compounds bind within the modulator-binding pocket and that substituents of each compound overlap with distinct moieties of cyclothiazide and CX614. The goals of the present study were to determine 1) the degree of modulation by CMPDA and CMPDB of AMPA receptor deactivation and desensitization; 2) whether these compounds are splice isoform-selective; and 3) whether predictions of mechanism of action could be inferred by comparing molecular interactions between the ligand-binding domain and each compound with those of cyclothiazide and CX614. CMPDB was found to be more isoform-selective than would be predicted from initial binding assays. It is noteworthy that these new compounds are both more potent and more effective and may be more clinically relevant than the AMPA receptor modulators described previously.


[Marriage of blood relatives and congenital deafness].
Audiological and genealogical examination of 658 patients with congenital deafness. All of them were citizens of the Republic of Azerbaijan. It was found that 236 (35.8%) probands had a family history of blood relatives marriage with a high inbreeding coefficient (F=0.0105). Marriages between cousin sibs by a parallel type prevailed. Marriages between blood relatives promote a rise in homozygosis of the pathological recessive genes increasing the risk of beath of handicapped babies.


What do parents expect antenatally and do babies teach them?
Antenatal inquiry of 658 parents in the North Staffordshire District Health Authority showed at least 13% to lack knowledge about a baby's potential for personal interaction when under 1 week old. Mothers and fathers varied significantly, independent of parity. Experienced parents expected later smiles. Two hundred and seventy eight of the same families were questioned three to four months after birth. Most parents had observed their baby's early responsiveness. Specific responses (Looking, listening) were noted earlier when anticipated and looked for. Significant among postnatal non-responders was their antenatal selection of predominantly impersonal stimulation for a baby. The greatest degree of warmth noted in postnatal observations correlated notably with a father's antenatal interest and postnatal support from him and their friends, a mother's commitment to breast feeding and her knowledge that a very young baby can see, like faces, and cries for emotional reasons. This highlights areas for antenatal tuition and postnatal encouragement, aiming to enhance warm early mutual regard between parents and child.


Safety and efficacy of subcutaneous Campath-1H for treating residual disease in patients with chronic lymphocytic leukemia responding to fludarabine.
Recent observations suggested that targeted monoclonal antibodies might be best employed in lymphoid malignancies under conditions of minimal residual disease. This prompted us to investigate the role of Campath-1H as treatment for patients with chronic lymphocytic leukemia (CLL) in whom fludarabine (FAMP) had produced a marked disease debulking with persistence of bone marrow (BM) infiltration or a complete remission (CR) without the disappearance of the molecular aberration (IgH monoclonal expression). As intravenous Campath-1H is almost invariably associated with reactions, sometimes of WHO grade 3-4, we adopted the subcutaneous route of administration, which proved to induce rare and mild adverse reactions but had comparable efficacy. DESIGN AND METHODS. Nine patients (7 males, 2 females) with a median age of 55 years (range 41-61) who responded to FAMP (1 had a CR, 5 a nodular partial remission [PRN], and 3 a partial remission [PR]), according to NCI Working Group Criteria, received subcutaneous Campath-1H, three times a week for 6 weeks in escalating doses up to 10 mg. Monoclonal rearrangement of IgH was present in all patients before immunotherapy. Patients received acyclovir and cotrimoxazole as infection prophylaxis. Granulocyte colony-stimulating factor (G-CSF), at the dosage of 5-10 microg/kg/die, or intermediate-dose Ara-C (800 mg/m(2)/q 12h x 6 doses), was administered to obtain peripheral blood stem cell (PBSC) mobilization. All patients were evaluable for response. Five patients, 2 in PR and 3 in PRN after FAMP treatment, reached a CR. Three patients, one in PR, one in PRN and one in CR, converted to a molecular remission. In four out of seven patients PBSC harvesting was successful; more than 2.5 x 10(6) cells/kg were collected from all these patients. Collection was polyclonal for IgH gene rearrangement in three cases. One patient has been transplanted after cyclophosphamide and total body irradiation as conditioning regimen, without complications and with rapid hemopoietic engraftment. All patients were evaluable for toxicity. A WHO grade 1-2 skin reaction was observed in 5 patients at the site of injection. No infectious episodes were recorded. Two out of three patients presenting cytomegalovirus reactivation, without pneumonia, were successfully treated with oral gancyclovir. Subcutaneous Campath-1H administered to CLL patients with residual BM disease after FAMP proved to be safe and effective. Of nine patients, three obtained a molecular CR and five converted into a morphologic and immunophenotypic CR. In four of seven patients submitted to PBSC mobilization, this treatment also allowed a harvest uncontaminated by CD5/CD19 double-positive CLL cells, which was polyclonal for IgH gene rearrangement in three cases.


Single application of a fluorescent test cream by healthy volunteers: assessment of treated and neglected body sites.
Management of dermatological self-treatment is demanding. Imperfect application of creams and ointments and poor adherence to topical treatment are common, resulting in unsatisfactory treatment outcome. To assess the technique and precision of test subjects' self-application of a test cream. Treated and neglected skin sites were measured after intended widespread single application of a fluorescent test cream. Twenty healthy volunteers (10 women, 10 men) were included. They were asked to treat their whole skin surface with the fluorescent test cream, except the head and neck and skin covered by underwear. Treated and untreated sites were subsequently measured under Wood's ultraviolet radiation. Thirty-one per cent of the skin surface that was a target for application did not show any fluorescence and thus was assumed to have been untreated. Typical neglected sites included the central back, the upper breast, the axilla with surrounding skin, the legs and the feet, particularly the sole. The posterior aspect of both trunk and extremities, not easily inspected, was more often neglected. In the treated sites the fluorescence was typically uneven. Qualified and motivated persons with no obvious physical limitations practised imperfect self-application of a test cream mimicking a therapeutic cream product. As much as 31% of the skin surface was neglected. Sites especially prone to nonapplication were identified. This might imply that dermatological patients on long-term self-treatment may practise local application very poorly, a problem of major therapeutic and economic importance. A fluorescent test cream can be used for research, and as an educational tool in the training of dermatological patients on how to apply local treatment.


Potential for differentiation of pseudoprogression from true tumor progression with dynamic susceptibility-weighted contrast-enhanced magnetic resonance imaging using ferumoxytol vs. gadoteridol: a pilot study.
We evaluated dynamic susceptibility-weighted contrast-enhanced magnetic resonance imaging (DSC-MRI) using gadoteridol in comparison to the iron oxide nanoparticle blood pool agent, ferumoxytol, in patients with glioblastoma multiforme (GBM) who received standard radiochemotherapy (RCT). Fourteen patients with GBM received standard RCT and underwent 19 MRI sessions that included DSC-MRI acquisitions with gadoteridol on Day 1 and ferumoxytol on Day 2. Relative cerebral blood volume (rCBV) values were calculated from DSC data obtained from each contrast agent. T1-weighted acquisition post-gadoteridol administration was used to identify enhancing regions. In seven MRI sessions of clinically presumptive active tumor, gadoteridol-DSC showed low rCBV in three and high rCBV in four, whereas ferumoxytol-DSC showed high rCBV in all seven sessions (p = 0.002). After RCT, seven MRI sessions showed increased gadoteridol contrast enhancement on T1-weighted scans coupled with low rCBV without significant differences between contrast agents (p = 0.9). Based on post-gadoteridol T1-weighted scans, DSC-MRI, and clinical presentation, four patterns of response to RCT were observed: regression, pseudoprogression, true progression, and mixed response. We conclude that DSC-MRI with a blood pool agent such as ferumoxytol may provide a better monitor of tumor rCBV than DSC-MRI with gadoteridol. Lesions demonstrating increased enhancement on T1-weighted MRI coupled with low ferumoxytol rCBV are likely exhibiting pseudoprogression, whereas high rCBV with ferumoxytol is a better marker than gadoteridol for determining active tumor. These interesting pilot observations suggest that ferumoxytol may differentiate tumor progression from pseudoprogression and warrant further investigation.


[Damage to the Ca2+-transport function and the lipid component of the sarcoplasmic reticulum membrane in total ischemia of the rat myocardium].
The Ca2+-transporting activity, lipoperoxide chemiluminescence and phospholipid spectrum of sarcoplasmic reticular membranes were studied in ischemic rats. It was shown that a substantial reduction in Ca2+ uptake rate by the sarcoplasmic reticulum occurred within the first 30 minutes and correlated with the increase in chemiluminescence intensity and accumulation of lysophosphatidylcholine. It has been suggested that free radical lipid peroxidation and phospholipase activation are directly related to the reduction of Ca2+-transporting rate by sarcoplasmic reticulum in myocardial ischemia.


B Cells Drive Autoimmunity in Mice with CD28-Deficient Regulatory T Cells.
Follicular regulatory T (TFR) cells are a newly defined regulatory T cell (Treg) subset that suppresses follicular helper T cell-mediated B cell responses in the germinal center reaction. The precise costimulatory signal requirements for proper TFR cell differentiation and function are still not known. Using conditional knockout strategies of CD28, we previously demonstrated that loss of CD28 signaling in Tregs caused autoimmunity in mice (termed CD28-ΔTreg mice), characterized by lymphadenopathy, accumulation of activated T cells, and cell-mediated inflammation of the skin and lung. In this study, we show that CD28 signaling is required for TFR cell differentiation. Treg-specific deletion of CD28 caused a reduction in TFR cell numbers and function, which resulted in increased germinal center B cells and Ab production. Moreover, residual CD28-deficient TFR cells showed a diminished suppressive capacity as assessed by their ability to inhibit Ab responses in vitro. Surprisingly, genetic deletion of B cells in CD28-ΔTreg mice prevented the development of lymphadenopathy and CD4+ T cell activation, and autoimmunity that mainly targeted skin and lung tissues. Thus, autoimmunity occurring in mice with CD28-deficient Tregs appears to be driven primarily by loss of TFR cell differentiation and function with resulting B cell-driven inflammation.


Foxa2 attenuates steatosis and inhibits the NF-κB/IKK signaling pathway in nonalcoholic fatty liver disease.
Forkhead box a2 (Foxa2) is proven to be an insulin-sensitive transcriptional regulator and affects hepatic steatosis. This study aims to investigate the mechanism by which Foxa2 affects nonalcoholic fatty liver disease (NAFLD). Animal and cellular models of NAFLD were constructed using high-fat diet (HFD) feeding and oleic acid (OA) stimulation, respectively. NAFLD mice received tail vein injections of either an overexpressing negative control (oe-NC) or Foxa2 (oe-Foxa2) for four weeks. HepG2 cells were transfected with oe-NC and oe-Foxa2 for 48 h before OA stimulation. Histological changes and lipid accumulation were assessed using hematoxylin-eosin staining and oil red O staining, respectively. Expression of Foxa2, NF-κB/IKK pathway proteins, lipid synthesis proteins, and fatty acid β-oxidation protein in HFD mice and OA-induced HepG2 cells was detected using western blot. Foxa2 expression was downregulated in HFD mice and OA-induced HepG2 cells. Foxa2 overexpression attenuated lipid accumulation and liver injury, and reduced the levels of aspartate aminotransferase, alanine aminotransferase, total cholesterol, or triglyceride in HFD mice and OA-induced HepG2 cells. Moreover, Foxa2 overexpression decreased the expression of lipid synthesis proteins and increased fatty acid β-oxidation protein expression in the liver tissues. Furthermore, overexpression of Foxa2 downregulated the expression of p-NF-κB/NF-κB and p-IKK/IKK in OA-induced HepG2 cells. Additionally, lipopolysaccharide (NF-κB/IKK pathway activator) administration reversed the downregulation of lipid synthesis proteins and the upregulation of fatty acid β-oxidation protein. Foxa2 expression is downregulated in NAFLD. Foxa2 ameliorated hepatic steatosis and inhibited the activation of the NF-κB/IKK signaling pathway.