A simple approach for multicolor immunofluorescence staining in different Drosophila cell types.
Multicolor immunostaining analysis is often a desirable tool in cell biology for most researchers. Nonetheless, this is not an easy task and often not affordable by many laboratories as it might require expensive instrumentation and sophisticated analysis software. Here, we describe a simple protocol for performing sequential immunostainings on two different Drosophila specimens. Our strategy relies on an efficient and reproducible method for removal primary antibodies and/or fluorophore-conjugated secondary antibodies that does not affect antigene integrity. We show that alternation of multiple rounds of antibody incubation and removal on the same slide, followed by registration of the same DAPI-stained image, provides a simple framework for the sequential detection of several antigens in the same cell. Given that the sample fixation procedures used for Drosophila tissues are compatible with most specimen processing protocols, we can envisage that the multicolor immunostaining strategy presented here can be also adapted to different samples including mammalian tissues and/or cells.


Abnormal organogenesis of Peyer's patches in mice deficient for NF-kappaB1, NF-kappaB2, and Bcl-3.
Nuclear factor (NF) kappaB1, NF-kappaB2, and Bcl-3 encode for proteins of the NF-kappaB/Rel/IkappaB families, known as regulators of innate and adoptive immune responses. Targeted disruption of these genes showed essential roles in lymphoid organ development and organization. NF-kappaB1-, NF-kappaB2-, and Bcl-3-deficient mouse lines were established, and their role in organogenesis of Peyer's patches (PP) was investigated. Macroscopic inspection showed a reduced number and size of PP in Bcl-3(-/-) and NF-kappaB1(-/-) mice but failed to detect PP in NF-kappaB2(-/-) mice. Whole-mount in situ hybridization revealed the presence of interleukin-7 receptor-alpha spots in NF-kappaB2(-/-) mice, indicating no defect in PP organogenesis of NF-kappaB2(-/-) mice in principle. Immunostaining shows that residual lymphocytes mainly consist of T cells. B cells are substantially reduced and are accumulated as terminal extravasations. Organized follicular structures and follicular dendritic cell networks fail to form, and myeloid, but not lymphoid, dendritic cells are obviously reduced. Expression of the chemokines macrophage inflammatory protein-3alpha, B-lymphocyte chemoattractant, and thymus-expressed chemokine is impaired in epithelial cells and in the subendothelial dome area that is not well defined. A similar but less severe phenotype is seen in Bcl-3(-/-) mice, which also do not develop germinal centers. In contrast, in NF-kappaB1(-/-) mice, T-cell numbers are visibly reduced, and no alteration could be observed in the B-cell and dendritic-cell populations. These data show that all 3 genes are crucial for PP development but contribute differently to PP organogenesis.


In vitro endocytosis of benign and malignant human bone marrow plasma cells.
We report ultrastructural evidence of the phagocytic potential of plasma cells and myeloma cells. The incubation of plasma cells and myeloma cells in vitro with horseradish peroxidase (HRP) and cationized ferritin (CF) allows the tracing of fluid-phase and receptor-mediated pathways. Surface-bound ligands (CF) and solutes (HRP) taken up in primary pinocytic vesicles are internalized to the endosomal compartment. After 1 hr of incubation, CF was found not only in plasma cells but also in myeloma cells. Reaction products of HRP were observed only in myeloma cells. In myeloma cells, however, HRP was located only in the lysosomal system, whereas CF was present within membrane cisternae as well as within lysosomes. These myeloma cells morphologically produced interleukin-6 (IL-6).


[CLINICAL, MORPHOLOGICAL AND MOLECULAR GENETIC FEATURES AND PROGNOSTIC FACTORS OF PRIMARY MYELOFIBROSIS. A CASE OF PRIMARY MYELOFIBROSIS TRANSFORMATION INTO ACUTE MYELOID LEUKEMIA].
Primary myelofibrosis is a common disease from the group of Ph-negative myeloproliferative diseases. The article presents modern data on the pathogenesis of Ph-negative myeloproliferative diseases, as well as diagnostic criteria, treatment tactics and prognosis factors for primary myelofibrosis. A clinical case of transformation of primary myelofibrosis into acute myeloid leukemia is described. Purpose of the study - to present up-to-date information on the pathogenesis, diagnostic criteria, principles of treatment and prognostic factors of primary myelofibrosis, as well as to present a clinical case of transformation of primary myelofibrosis into acute myeloblastic leukemia. According to modern concepts, for the early diagnosis of primary myelofibrosis, along with the clinical and morphological methods of examining patients, molecular genetic verification of the disease is extremely important. To improve the survival rate of patients with primary myelofibrosis, molecular genetic verification of the disease and stratification for the choice of treatment tactics are necessary.


Changes in Simple Spirometric Parameters After Lobectomy for Bronchial Carcinoma.
The purpose of this study was to describe the postoperative changes in lung function after pure open lobectomy for lung carcinoma. 30 patients (mean age 64 ± 7 years old, 16 men and 14 women) underwent a left or right lobectomy. They underwent spirometric pulmonary tests preoperatively, and at 1 and 6 months after the operation. The average preoperative forced expiratory volume in 1 second (FEV1) was 2.55±0.62lt and the mean postoperative FEV1 at 1 and 6 months was 1.97 ± 0.59 L and 2.15±0.66 L respectively. The percentage losses for FEV1 were 22.7% and 15.4% after 1 and 6 months respectively. An average percentage increase of 9.4% for FEV1 was estimated at the time of 6 months in comparison with this of 1 month after the operation. The average preoperative forced vital capacity (FVC) was 3.17 ± 0.81 L and the mean postoperative FVC at 1 and 6 months after the operation was 2.50 ± 0.63 L and 2.72 ± 0.67 L respectively. The percentage losses for FVC were 21.1% and 14.2% after 1 and 6 months respectively. An average percentage increase of 8.7% was observed at the time period of 6 months in comparison with this of 1 month after the operation. Although, we observed a significant decrease in FEV1 and FVC after the operation, all patients were in excellent clinical status. FEV1 and FVC of 6 months were increased in comparison with the respective values of 1 month after the operation, but did not reach the preoperative values in any patient.


Enzymic nature of the protein moiety of protochlorophyllide holochrome.
The enzymic nature of the protein moiety of protochlorophyll(ide) holochrome was studied by following the fate of the [(14)C]protochlorophyll(ide) formed when dark-grown barley (Hordeum vulgare) or bean (Phaseolus vulgaris) leaves are incubated in the dark with 3 mm 4-delta-[(14)C]aminolevulinic acid. It was found that: [List: see text]Since turnover of protochlorophyll(ide) was not observed, these results show that there is a free exchange between the old "endogenous" and the new delta-aminolevulinic-acid-induced protochlorophyll(ide) molecules on the active site of the holochrome protein. These results are consistent with the hypothesis that the holochrome protein acts as an enzyme.


Primitive hemopoietic stem cells: direct assay of most productive populations by competitive repopulation with simple binomial, correlation and covariance calculations.
Quantitative analyses of primitive hemopoietic stem cell (PHSC) populations are important both for basic biology and for clinical applications. Unfortunately, many conventional assays fail to measure long-term repopulating ability and maximal differentiating ability, the most important characteristics of the PHSC. The competitive repopulation assay described here focuses on this characteristic, assaying the precursors from which most differentiated cells are descended over large fractions of the life span in laboratory mice. Thus long-term repopulating ability and the ability to differentiate into both myeloid and lymphoid lineages are measured directly from 2.5 to 12.5 months after transplantation. This technique also has found high correlations between granulocytes, macrophages, and T and B lymphocytes as early as 3 weeks after transplantation. All or most differentiated cells of these widely disparate types appear to be descended from a common precursor cell, while myeloid-specific or lymphoid-specific precursors produce few or no descendants. However, large increases in variances between 3 and 6 weeks and 12 weeks after transplantation suggest that most of the initially active multilineage precursors are exhausted. Thus the ability to differentiate into widely disparate lineages does not establish long-term repopulating ability.


Are we ignoring neutral and negative human-animal relationships in zoos?
Human-animal interactions (HAI), which may lead to human-animal relationships (HAR), may be positive, neutral, or negative in nature. Zoo studies show that visitors may be stressful, may have no effect, or may be enriching. There is also evidence that good HARs set up between animals and their keepers can have positive effects on animal welfare. However, we need to know more about negative HARs, and as a first step we attempt to do this here by considering cases where animals attack people in the zoo. Due to the sensitivity and rarity of these events data appear sparse and unsystematically collected. Here, information available in the public domain about the circumstances of these attacks has been collated to test hypotheses about negative HAIs derived from a model of zoo HARs. The limited data presented here broadly support the zoo HAR model, and suggest that attacks usually happen in unusual circumstances, where there may be a failure by the animal to recognise the HAR, or where the relationship, if there is one, does not hold; and give some support to the prediction that exposure to many keepers may impair the development of a positive HAR. This study may provide useful information for the zoo community to proactively collect systematic standardised records, which will enable a fuller understanding of zoo HARs, upon which appropriate measures might be adopted to build better zoo HARs, which are likely to positively impact zoo animal welfare, and reduce these rare incidences further.


Overexpression of Arabidopsis CBF1 gene in transgenic tobacco alleviates photoinhibition of PSII and PSI during chilling stress under low irradiance.
A known arabidopsis cDNA clone, the CRT/DRE binding factor 1 (CBF1), was isolated and introduced into tobacco plants. It has been reported that CBF1 is one member of CBF gene family related to low temperature and enhancing low temperature tolerance of plants. In the present work, the transcripts could be detected in the transgenic lines. The photochemical efficiency of PSII (F(v)/F(m)) and the photo-oxidizable P700 in the transgenic lines overexpressing CBF1 were higher than that in the wild type plants during the chilling stress under low irradiance. Similarly, the higher NPQ, higher qf, lower Phi(NF), higher activity of SOD, and lower content of MDA were also detected in the transgenic tobacco lines. Additionally, higher expression levels of Nicotiana tabacum copper/zinc superoxide dismutase (Cu/Zn SOD) were also detected in the transgenic lines. These results suggest that CBF1 protein plays an important role in protection of PSII and PSI during the chilling stress under low irradiance.


Alternative long-ranged charge optimized many-body potential for aluminium.
A new COMB3 potential was developed for aluminium, which focuses on long-range interaction and phase transition. The potential was developed by fitting the equilibrium lattice properties of different phases and defects to ensure its transferability to general systems. The quality of the potential was tested in several problems and compared with the EAM potential as well as the published COMB3 potential, the effect of the cutoff method was studied in detail to demonstrate the necessity to extend the cutoff region. Systems of strong deformations along the Bain path, under a trigonal strain and with planar stacking faults were calculated and the present potential performed as well as the EAM potential. At last, a surface process that involves adsorption and diffusion was studied using the present potential.


Stenting treatment for intra- and extra-cranial atherosclerotic diseases.
The purpose of this study was to investigate the effect, safety and short-term outcome of stenting treatment for extra- and intra-cranial atherosclerotic diseases. Forty-one patients underwent stenting treatment using coronary and biliary stents. Eleven patients had tubular or diffuse high-grade stenoses not amenable to standard balloon angioplasty involving intra-cranial arteries, while in thirty, extra-cranial arteries were involved. Stents were successfully implanted in 36 out of 41 lesions (88%) with 2% (1/41) of overall procedural morbidity rate. Procedural and clinical success rate of intra-cranial stent placement was 64% (7/11) and no complications occurred during or after intra-cranial stent placement: the morbidity rate was 0%. Hyperperfusion injury occurred in two patients after successful implantation of stents in subtotal occlusion of the internal carotid artery, and consequently the overall clinical success rate was 80% (33/41). Restenosis occurred in four (12%) out of 33 patients who underwent six month follow-up arteriography. No ischemic attacks or stent-deformation occurred during follow-up after stenting treatment. For lesions not amenable to standard balloon angioplasty, endovascular stent placement seems to be a safe and effective treatment of modality.


Uptake of a natural surfactant and increased delivery of small organic anions into type II pneumocytes.
The uptake of natural lung surfactant into differentiated type II cells may be used for the targeted delivery of other molecules. The fluorescent anion pyranine [hydroxypyren-1,3,6-trisulfonic acid, sodium salt (HPTS)] was incorporated into a bovine surfactant labeled with [3H]dipalmitoylphosphatidylcholine ([3H]DPPC). The uptake of [3H]DPPC and of HPTS increased with time of incubation and concentration, decreased with the size of the vesicles used, and was stimulated by 8-bromo-cAMP and partially inhibited by hypertonic sucrose. However, the amount of HPTS uptake was approximately 100 times smaller than that of [3H]DPPC. This large difference was due to a more rapid regurgitation of some of the HPTS from the cells but not to leakage from the surfactant before uptake. The acidification of the internalized surfactant increased linearly over 90 min to 7.13, and after 24 h, a pH of 6.83 was measured. In conclusion, after internalization of a double-labeled natural surfactant, the lipid moieties were accumulated in relation to the anions, which were targeted to a compartment not very acidic and in part rapidly expelled from the cells.


Population policies and the "creation" of Africa.
"This paper will look at the ways in which population policies have played an important role in the ongoing debate about African identities and culture.... In dealing with the construction of Africa, this paper will be limited to the impact of the population discourse on Africans. Three groups will be considered. These are: the target population of (largely rural) women; males (representing the patriarchal dimension); and African intellectuals. Even though membership in each group is not mutually exclusive, each will be dealt with separately for ease of analysis." (SUMMARY IN FRE)


Geometric Analysis of a System with Chemical Interactions.
In this paper, we present some initial results aimed at defining a framework for the analysis of thermodynamic systems with additional restrictions imposed on the intensive parameters. Specifically, for the case of chemical reactions, we considered the states of constant affinity that form isoffine submanifolds of the thermodynamic phase space. Wer discuss the problem of extending the previously obtained stability conditions to the considered class of systems.


Trends in mammography use and their relation to physician and other factors.
This study examines trends and factors (e.g., physician utilization) affecting mammography use from 1988 to 1990 among women residing in an area of high breast cancer incidence. Mail surveys of independent random samples of over 2000 women 50 to 75 years of age residing on Long Island were conducted each of the three study years, and responses were compared. Statistically significant increases in reported mammography use occurred over the 3-year period within all age, income, and educational subgroups. These were accompanied by a decline in reporting that the doctor did not recommend it, as a reason for not having a mammogram, and by a statistically significant increase in reported physician recommendation among those who visit a doctor annually, particularly those using a gynecologist. Clinical breast examination was the strongest predictor of mammography use, with knowledge of the recommended screening frequency also strong associated. Despite a regionally high incidence of breast cancer, screening mammography use corresponded to national rates. Access and visits to a regular physician were critical factors and substantiated the need for education of women, family practitioners, and internists about breast cancer screening guidelines.


[Gastric MALT lymphoma: recent topics regarding treatment and genetic aberrations].
The recent topics in the treatment and genetic aberrations of gastric MALT lymphoma were reviewed. The chromosomal translocation t(11;18) (q21;q21)/API2-MALT1 has been frequently detected not only in gastric MALT lymphoma cases, but also in gastric diffuse large B-cell lymphoma (DLBCL) cases in two studies. Conversely, IGH-involved translocations, including t(3;14) (p14;q32)/FOXP1-IGH, are rarely seen in gastric MALT lymphoma. Several large-scaled clinical series have revealed that Helicobacter pylori eradication therapy is the optimal first-line treatment for patients with gastric MALT lymphoma, which leads to a favorable long-term outcome. A number of modalities and/or regimens, including chemotherapy, immunotherapy, and radioimmuotherapy, have been developed as the second-line treatments for patients not responding to eradication therapy.


Characterization of Ras effector mutant interactions with the NF1-GAP related domain.
The GTPase activating proteins Ras GAP and the neurofibromatosis-type 1 (NF1) gene product have been implicated as both potential mediators and regulators of the mitogenic effects of the ras proteins. In this study, the interactions of selected Ras effector mutants with the NF1-GAP related domain (NF1-GRD) were investigated. The NF1-GRD was unable to stimulate the GTPase of Ras[Asn33], Ras[Ser35] or Ras[Asn38], all transformation defective mutants. Each of these mutants had reduced but detectable binding to the NF1-GRD (apparent KD of 9 microM, 4 microM and 2 microM respectively, vs 0.5 microM for normal Ha-ras). The NF1-GRD was able to fully stimulate the intrinsic GTPase of the transformation-defective Ras[Gly26Ile27] and Ras[Glu45] mutants, each of which bound the NF1-GRD with wild type affinity or better (KD = 0.13 microM and 0.4 microM respectively). The transforming Ras[Glu30Lys31] protein showed no GTPase stimulation and bound most poorly to the NF1-GRD (apparent KD of 16 microM). The interaction of the NF1-GRD with these specific Ras effector mutations is similar to that observed for Ras GAP. When the relative transforming activity of the valine 12 form of each Ras mutant was plotted against the apparent KD for NF1-GRD binding, little correlation was observed. These results support the hypothesis that the NF1 gene product does not function as a downstream effector of Ras in the mitogenic pathway.


Neurosecretion competence. A comprehensive gene expression program identified in PC12 cells.
The phenotype of neurosecretory cells is characterized by clear vesicles and dense granules, both discharged by regulated exocytosis. However, these organelles are lacking completely in a few neurosecretion-incompetent clones of the pheochromocytoma PC12 line, in which other specific features are maintained (incompetent clones). In view of the heterogeneity of PC12 cells, a differential characterization of the incompetent phenotype based on the comparison of a single incompetent and a single wild-type clone would have been inconclusive. Therefore, we have compared two pairs of PC12 clones, studying in parallel the transcript levels of 4,200 genes and 19,000 express sequence tags (ESTs) by high density oligonucleotide arrays. After accurate data processing for quality control and filtration, a total of 755 transcripts, corresponding to 448 genes and 307 ESTs, was found consistently changed, with 46% up-regulated and 54% down-regulated in incompetent versus wild-type clones. Many but not all neurosecretion genes were profoundly down-regulated in incompetent cells. Expression of endocytosis genes was normal, whereas that of many nuclear and transcription factors, including some previously shown to play key roles in neurogenesis, was profoundly changed. Additional differences appeared in genes involved in signaling and metabolism. Taken together these results demonstrate for the first time that expression of neurosecretory vesicles and granules is part of a complex gene expression program that includes many other features that so far have not been recognized.


Study on the Mechanism of Huanglian Jiedu Decoction in Treating Dyslipidemia Based on Network Pharmacology.
This study aimed to determine the active ingredients of Huanglian Jiedu decoction (HLJDD) and the targets for treating dyslipidemia through network pharmacology to facilitate further application of HJJDD in the treatment of dyslipidemia. Potential drug targets for dyslipidemia were identified with a protein-protein interaction network. Gene ontology (GO) enrichment analysis and KEGG pathway analysis were performed to elucidate the biological function and major pathways involved in the HLJDD-mediated treatment of dyslipidemia. This approach revealed 22 components, 234 targets of HLJDD, and 221 targets of dyslipidemia. There were 14 components and 31 common targets between HLJDD and dyslipidemia treatment. GO enrichment analysis showed that these targets were mainly associated with the response to DNA-binding transcription factor activity, lipid localization and storage, reactive oxygen species metabolic process, and inflammatory response. The results of KEGG analysis indicated that the AGE-RAGE, NF-κB, HIF-1, IL-17, TNF, FoxO, and PPAR signalling pathways were enriched in the antidyslipidemic action of HLJDD. This study expounded the pharmacological actions and molecular mechanisms of HLJDD in treating dyslipidemia from a holistic perspective, which may provide a scientific basis for the clinical application of HLJDD.


Genome-Wide Identification, Characterization and Expression Analysis of Lipoxygenase Gene Family in Artemisia annua L.
Lipoxygenase (LOX) is a ubiquitous oxygenase found in animals and plants and plays a pivotal role in diverse biological processes, including defense and development. Artemisinin, which can only be obtained from Artemisia annua L., is the most effective therapeutic drug for malaria without serious side effects. This study identified and analyzed LOX gene family members in the A. annua genome at the chromosomal level. Twenty LOX genes with various molecular weights, isoelectric points, and amino acid numbers were identified and named AaLOX, which were located in the cytoplasm or chloroplast. The average protein length of all AaLOX was 850 aa. Phylogenetic tree analysis revealed that the AaLOX was divided into two major groups, 9-LOX and 13-LOX. The exon numbers ranged from 1 to 12, indicating that different AaLOX genes have different functions. The secondary structure was mainly composed of alpha helix and random coil, and the tertiary structure was similar for most AaLOX. Upstream promoter region analysis revealed that a large number of cis-acting elements were closely related to plant growth and development, light response, hormone, and other stress responses. Transcriptome data analysis of different tissues suggested that the gene family was differently expressed in the roots, stems, leaves, and flowers of two A. annua strains HAN1 and LQ9. qRT-PCR confirmed that AaLOX5 and AaLOX17 had the highest expression in flowers and leaves. This study provides a theoretical basis for the further functional analysis of the AaLOX gene family.


There is no significant correlation of adenomyosis with benign, premalignant and malignant gynecological pathologies. Retrospective study on 647 specimens.
The aim of this study is to identify the prevalence of benign, premalignant and malignant gynecological pathologies in women with adenomyosis who underwent gynecological surgery. The medical records collected between 1985 and 2020 were retrospectively reviewed. The pathology reports were studied from 647 cases where adenomyosis was presented. The estimated prevalence of benign, premalignant and malignant gynecological disorders in the general population was further evaluated. The mean age of women with adenomyosis was 54.1 ± 10.4 years old. Out of 647 patients, in 18.5% of the specimens we detected isolated adenomyosis and in 81.5% of cases a coexistence of one or more gynecological diseases, while in 84 out of 647 patients (13%) there was coexistence of adenomyosis with more than one gynecological condition (benign or malignancy). Among all cases, uterine leiomyomas were observed in 61.3% of patients, followed by endometrial polyps (11.9%), endometriosis (11.6%), endometrial hyperplasia (7.1%), endometrial cancer (3.6%), ovarian (1.4%) and cervical cancer (0.8%) (p < 0.001).Additionally, we found that women with a simultaneous co-existence of adenomyosis, leiomyomas and endometrial polyps or hyperplasia were younger (p < 0.01) in comparison to cases with malignancy. Adenomyosis presents a common benign but often progressing myometrial condition that it is underestimated in clinical practice. Even though some studies suggest a potential association with several gynecological pathologies, we did not confirm a significant difference of adenomyosis prevalence between benign, premalignant and malignant gynecological conditions compared with the general population. Further investigation is required to confirm our results.


Role of Cancer Surgery in the Improvement of the Operative Skills of Military Surgeons During Deployment: A Single-Center Study.
The maintenance of military surgeons' operative skills is challenging. Different and specific training strategies have been implemented in this context; however, little has been evaluated with regard to their effectiveness. Cancer surgery is a part of military surgeons' activities in their home hospitals. This study aimed to assess the role of oncological surgery in the improvement of military surgeons' operative skills. Between January and June 2019, the surgical activities of the departments of visceral, ear, nose, and throat, urological, and thoracic surgery were retrospectively reviewed and assessed in terms of the operative time (OT). All surgeons working at the Sainte Anne Military Teaching Hospital were sent a survey to rate on a 5-point scale the current surgical practices on their usefulness in improving surgical skills required for treating war injuries during deployment (primary endpoint) and to compare on a 10-point visual analog scale the influence of cancer surgery and specific training on surgical fluency (secondary endpoint). Over the study period, 2,571 hours of OT was analyzed. Oncological surgery represented 52.5% of the surgical activity and almost 1,350 hours of cumulative OT. Considering the primary endpoint, the mean rating allocated to cancer surgery was 4.53 ± 0.84, which was not statistically different than that allocated to trauma surgery (4.42 ± 1.02, P = 0.98) but higher than other surgery (2.47 ± 1.00, P < 0.001). Considering the secondary endpoint, cancer surgery was rated higher than specific training by all surgeons, without statistically significant difference (positive mean score of + 2.00; 95% IC: 0.85-3.14). This study demonstrates the usefulness of cancer surgery in improving the operative skills of military surgeons.


Altered CD45 expression in malignant B-1 cells.
CD45 is an important surface glycoprotein which has an intrinsic tyrosine phosphatase activity and has been implicated in cell proliferation, signaling, and differentiation and is associated with the B cell receptor during signaling. In this manuscript, the role of CD45 expression in the development of B-1 malignancies in NZB mice, which serve as a model for human diseases such as chronic lymphocytic leukemia, was investigated. B-1 cells spontaneously hyperproliferate and form a clonal hyperdiploid malignant population in aging NZB mice. Phenotypic analysis indicates that the NZB malignant B-1 cells are bright for IgM, but have reduced levels of CD45 relative to normal, nonmalignant B cells (both B-1 and B-2) and are characterized by dull or negative expression of the CD45 isoform B220/6B2 normally found on all B cells. Malignant B-1 cells demonstrated decreased RNA levels of CD45 relative to IgM expression, while nonmalignant B-2 cells showed similar levels of RNA expression for both CD45 and IgM. As CD45 exists in several isoforms and B cells express the highest molecular weight isoform (B220), malignant B-1 cells were further analyzed with respect to their isoform usage. Although, at the RNA level malignant B-1 cells showed the presence of the of the B220 form of CD45, western blot analysis of B220 protein suggested a posttranslational glycosylation defect in the CD45/B220 expression recognized by the mAb 6B2. F1 recipients of premalignant NZB B-1 cells which had been sorted for IgMhi, B220/6B2negative cells developed hyperdiploid malignant donor B-1 clones earlier than did recipients of NZB B-1 cells which were bright for B220/6B2. However, all the malignant B-1 clones of NZB origin which developed in recipients of both transfer populations were B220/6B2 negative. This indicated that abnormal expression of CD45 may be prerequisite for long-term growth and malignant transformation. Thus alterations in CD45 may result in abnormal functioning of the malignant B-1 cells which may further affect the proliferation of, or signaling within, these cells.


Helminth products as a potential therapeutic strategy for inflammatory diseases.
Helminths secrete several molecules that can modulate the immune responses, favoring their evasion and perpetuate their survival in the host. These molecules interfere with antigen presentation, cell proliferation and activation, antibody production, cause cell death, and stimulate regulatory responses. Here, we focus on some helminth products and address their immunomodulatory effects in the host immune system and, also, we describe some anti-inflammatory properties of an Ascaris suum-derived immunomodulatory molecule, named PAS-1. This protein is a 200-kDa molecule isolated by affinity chromatography using MAIP-1 (monoclonal antibody which recognizes PAS-1), coupled to Sepharose 4B. It suppresses the inflammatory responses in murine models of delayed-type hypersensitivity, lung allergic inflammation and LPS-induced inflammation into air pouches. PAS-1 also stimulates the secretion of regulatory cytokines such as IL-10 and TGF-beta and primes IFN-gamma-secreting CD8+ and IL-10/ TGF-beta-secreting CD4+CD25+ cell clones that avoid the lung inflammation. Thus, this protein is a potent immunomodulatory component that may be used for therapeutic interventions in inflammatory diseases.


Effectiveness of peer-delivered Center for Independent Living supports for individuals with psychiatric disabilities: A randomized, controlled trial.
The goal of this study was to examine the effectiveness of peer-delivered core services of Centers for Independent Living (CILs), which include advocacy, information and referral, skills training, and peer support. Ninety-nine individuals with a schizophrenia-spectrum or affective disorder who identified at least 3 needs were recruited from mental health centers and randomly assigned to be contacted by a certified peer specialist at a local CIL (CIL condition) or services as usual (SAU condition). Data on community participation, recovery, empowerment, quality of life, and needs were obtained at baseline and 6 and 12 months postbaseline, along with responses to open-ended questions about supports received. Participation in CIL supports was very limited. No differences were found in repeated measures analyses (Time × Condition). Post hoc analyses did show some positive results for those in the CIL condition. More than half of CIL participants described obtaining a substantive support in at least 1 area, and almost half of these resulted in some tangible new resource. Engagement in CIL supports was very limited, as were outcomes. Nonetheless, numerous examples of supports across a broad range of areas were reported along with examples of how needs were met. CIL supports, which are widely available around the United States, may offer a unique philosophy and approach for addressing the needs of individuals with psychiatric disabilities and are deserving of additional study. (PsycINFO Database Record


Characterization and development of EST-SSR markers derived from transcriptome of yellow catfish.
Yellow catfish (Pelteobagrus fulvidraco) is one of the most important freshwater fish due to its delicious flesh and high nutritional value. However, lack of sufficient simple sequence repeat (SSR) markers has hampered the progress of genetic selection breeding and molecular research for yellow catfish. To this end, we aimed to develop and characterize polymorphic expressed sequence tag (EST)-SSRs from the 454 pyrosequencing transcriptome of yellow catfish. Totally, 82,794 potential EST-SSR markers were identified and distributed in the coding and non-coding regions. Di-nucleotide (53,933) is the most abundant motif type, and AC/GT, AAT/ATT, AAAT/ATTT are respective the most frequent di-, tri-, tetra-nucleotide repeats. We designed primer pairs for all of the identified EST-SSRs and randomly selected 300 of these pairs for further validation. Finally, 263 primer pairs were successfully amplified and 57 primer pairs were found to be consistently polymorphic when four populations of 48 individuals were tested. The number of alleles for the 57 loci ranged from 2 to 17, with an average of 8.23. The observed heterozygosity (HO), expected heterozygosity (HE), polymorphism information content (PIC) and fixation index (fis) values ranged from 0.04 to 1.00, 0.12 to 0.92, 0.12 to 0.91 and -0.83 to 0.93, respectively. These EST-SSR markers generated in this study could greatly facilitate future studies of genetic diversity and molecular breeding in yellow catfish.


TP53 status and impact on AML prognosis within the ELN 2022 risk classification.
This study reports the following: (1) frequency of TP53 comutation within each component of the European LeukemiaNet 2022 acute myeloid leukemia risk classification, (2) relevance of TP53 mutated variant allelic fraction <10%, (3) prognostic impact of -7, -5/del(5q), -17/abn(17p), complex karyotype/monosomal karyotype, or myelodysplasia-related gene mutations with/without mutated TP53.


Changes in lactate dehydrogenase on admission throughout the COVID-19 pandemic and possible impacts on prognostic capability.
Introduction: The enzyme lactate dehydrogenase (LDH) is a good marker of general hyperinflammation correlated with mortality for COVID-19, and is therefore used in prognosis tools. In a current COVID-19 clinical randomized trial (CRT), the blood level of LDH was selected as an inclusion criterion. However, LDH decreased during the pandemic; hence, the impact of this decrease on the prognostic value of LDH for mortality was evaluated. Methods: Data on LDH levels in 843 patients were obtained and analyzed. Relative risk, standard error and receiver operating characteristic curves were calculated for two cutoff values. Results: Relative risk lost validity and the area under the curve narrowed by trimester during the pandemic. Conclusion: The progressive decrease in LDH impacted the capacity to predict mortality in COVID-19. More studies are needed to validate this finding and its implications.


Acute regulation of lymphocyte beta 2-adrenoceptors is altered in patients with essential hypertension.
The effects of acute stimulation of the sympathetic activity by dynamic exercise on lymphocyte beta 2-adrenoceptor density [assessed by (-)-125iodocyanopindolol (ICYP) binding] and responsiveness [10 mumol/l isoprenaline-induced cyclic adenosine monophosphate (cAMP) increases] were studied in 10 normotensive (Pdiast < 90 mmHg) volunteers and in 10 patients with established essential hypertension (Pdiast > 95 mmHg). In normotensives, dynamic exercise on a bicycle (80% of maximum heart rate) for 15 min led to an increase in lymphocyte beta 2-adrenoceptor density from 1080 +/- 77 to 2033 +/- 152 ICYP binding sites/cell; concomitantly isoprenaline-induced increase in lymphocyte cAMP was enhanced. This effect appears to be mediated by beta 2-adrenoceptor stimulation, since the exercise-induced increase in beta 2-adrenoceptor density was markedly attenuated by pretreatment of the volunteers with propranolol (5 mg intravenously 45 min before exercise), but not by pretreatment with the beta 1-selective antagonist bisoprolol (2.5 mg intravenously 30 min before exercise). In patients with essential hypertension, lymphocyte beta 2-adrenoceptor density (1512 +/- 101 ICYP binding sites/cell) was significantly higher than in controls (P < 0.05); the same held true for isoprenaline-induced cAMP increases. In these patients, however, dynamic exercise caused only a slight increase in lymphocyte beta 2-adrenoceptor density (to 1859 +/- 154 ICYP binding sites/cell) and in isoprenaline-induced cAMP increases. From these results it is concluded that in essential hypertension acute regulation of the beta-adrenoceptor/adenylate cyclase system is impaired.


Differential Oxygen Response of Photosynthesis in Soybean and Panicum milioides.
The carbon dioxide compensation concentration of Panicum milioides was less than that of soybean over the range of 15 to 35 C. In soybean (Glycine max [L.] Merr. cv. Wayne), the compensation concentration was directly proportional to O(2) concentration. In P. milioides, the compensation concentration was near zero up to 10% O(2) and then increased linearly with higher O(2), although the slope of the response was less than that in soybean. Leaf extracts of P. milioides contained 3-fold higher phosphoenolpyruvate carboxylase activity than soybean leaf extracts. Oxygen inhibition of photosynthesis and carboxy-lation efficiency was less in P. milioides than that observed in soybean. The affinity of P. millioides ribulose-1,5-di-P carboxylase for CO(2) appeared to be slightly greater than that of soybean. The affinity of both enzymes for O(2) was similar. The reduced response of the compensation concentration and photosynthesis to O(2) in P. milioides may be explained by photosynthetic phosphoenolpyruvate carboxylase fixation and by an apparent increased affinity of ribulose-1,5-di-P carboxylase for CO(2).


Microtubule depolymerization by the Kinesin-8 motor Kip3p: a mathematical model.
Proteins from the kinesin-8 family promote microtubule (MT) depolymerization, a process thought to be important for the control of microtubule length in living cells. In addition to this MT shortening activity, kinesin 8s are motors that show plus-end directed motility on MTs. Here we describe a simple model that incorporates directional motion and destabilization of the MT plus-end by kinesin 8. Our model quantitatively reproduces the key features of length-versus-time traces for stabilized MTs in the presence of purified kinesin 8, including length-dependent depolymerization. Comparison of model predictions with experiments suggests that kinesin 8 depolymerizes processively, i.e., one motor can remove multiple tubulin dimers from a stabilized MT. Fluctuations in MT length as a function of time are related to depolymerization processivity. We have also determined the parameter regime in which the rate of MT depolymerization is length dependent: length-dependent depolymerization occurs only when MTs are sufficiently short; this crossover is sensitive to the bulk motor concentration.


Planar Hexacoordinate Carbons: Half Covalent, Half Ionic.
Herein, the first global minima containing a planar hexacoordinate carbon (phC) atom are reported. The fifteen structures belong to the CE3 M3 + (E=S-Te and M=Li-Cs) series and satisfy both geometric and electronic criteria to be considered as a true phC. The design strategy consisted of replacing oxygen in the D3h CO3 Li3 + structure with heavy and less electronegative chalcogens, inducing a negative charge on the C atom and an attractive electrostatic interaction between C and the alkali-metal cations. The chemical bonding analyses indicate that carbon is covalently bonded to three chalcogens and ionically connected to the three alkali metals.


Affection of Respiratory Muscles in ALS and SMA.
Respiratory dysfunction is a common cause of morbidity and mortality in motor neuron disease (MND). However, classical volitional measures of respiratory function in these patients are impeded by, e.g., bulbar paralysis or progressive disability. Diaphragm ultrasound imaging might be a valuable tool for assessing respiratory impairment, albeit different ultrasound measures have not been systematically investigated in adult MND patients and, in particular, have not yet been comparatively applied in adult patients with amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA). We hypothesized that in contrast to ALS patients, adult SMA patients show a relative sparing of diaphragm function. We retrospectively analyzed diaphragm ultrasound imaging data of 40 patients with ALS and 23 patients with SMA in comparison to a multitude of established parameters of respiratory function. Indeed, ALS patients showed more severe diaphragm dysfunction than adult SMA patients, however, diaphragm dysfunction was also common in adult SMA patients. Notably, dynamic measures of diaphragm function rather than thickness measures were impaired in ALS compared to SMA. Thus, diaphragm ultrasound imaging might be a useful tool to evaluate respiratory dysfunction in adult MND patients. Future larger and prospective studies are needed to validate our initial findings.


Acclimation of activated sludge to degrade toxic levels of 2,4-dinitrophenol.
Biodegradation of 75 and 100 mg/l of 2,4-dinitrophenol (DNP) by activated sludge acclimated in a Sequencing Batch Reactor (SBR) consistently required less than 6 hours although a lag at the beginning of every 48-hour SBR cycle was observed. Other investigators have reported that DNP levels of 100 mg/l and higher are significantly toxic even to acclimated bacteria. The activated sludge acclimated to 75 mg/l initial DNP had over 100 times the DNP-degrading bacteria than an SBR acclimated to 10 mg/l DNP, although the MLSS concentration in both reactors was similar. Results suggest that two mechanisms are responsible for activated sludge acclimation to toxic levels of DNP: maintenance of DNP-degrading biomass sufficiently large to reduce initial DNP to non-toxic levels, allowing for subsequent rapid degradation; and extension of the aeration period well beyond the time required for degradation to prevent gradual accumulation of any by-product which might also be toxic.


Distal Transradial Access (dTRA) for Coronary Angiography and Interventions: A Quality Improvement Step Forward?
Distal TRA is a novel access site in the interventional cardiology field, with current data demonstrating high success and infrequent complication rates - global procedural metrics that are comparable with historical proximal TRA. Distal TRA may conversely provide important advantages over proximal TRA, including patient comfort, shorter hemostasis time, and lower radial artery occlusion rates.


The Antibacterial and Antibiofilm Activity of Telithromycin Against Enterococcus spp. Isolated From Patients in China.
Telithromycin has been reported to possess robust in vitro antibacterial activity against many species of gram-positive bacteria, and telithromycin is also effective against Staphylococcus aureus biofilms. However, the in vitro antimicrobial susceptibility of telithromycin against clinical enterococci isolates in China is rarely reported and the impacts of telithromycin on the biofilm formation and eradication of enterococci remain elusive. Therefore, this study aimed to explore the inhibitory effects of telithromycin on planktonic cells and biofilms of Enterococcus strains. A total of 280 Enterococcus faecalis and 122 Enterococcus faecium isolates were collected from individual inpatients in China. The 50% minimum inhibitory concentration (MIC50) values of telithromycin against the E. faecalis and E. faecium strains carrying erythromycin-resistant methylase (erm) genes such as the ermA, ermB, or ermC, were 2 and 4 μg/mL, respectively. In addition, these isolates were typed using multilocus sequence typing (MLST) based on housekeeping genes. The predominant sequence types (STs) of E. faecalis were ST16, ST30, and ST179, and the main STs of E. faecium isolates were ST18, ST78, and ST80. Among these major STs, 87.1% (135/158) of E. faecalis and 80.4% (41/51) of E. faecium carried erm genes. Furthermore, at the subinhibitory concentrations (1/4 and 1/8 × MIC) of telithromycin, the biofilm formation of 16 E. faecalis isolates were inhibited by approximately 35%. Moreover, treatment with 8 × MIC of telithromycin or ampicillin led to an almost 40% reduction in the established biofilms of E. faecalis isolates, whereas vancomycin or linezolid with 8 × MIC had minimal effects. The combination of telithromycin and ampicillin resulted in an almost 70% reduction in the established biofilms of E. faecalis. In conclusion, these results revealed that telithromycin significantly decreased the planktonic cells of both E. faecalis and E. faecium. In addition, the data further demonstrated that telithromycin has the robust ability to inhibit E. faecalis biofilms and the combination of telithromycin and ampicillin improved antibiofilm activity. These in vitro antibacterial and antibiofilm activities suggest that telithromycin could be a potential candidate for the treatment of enterococcal infections.


Ethanol-Enriched Substrate Facilitates Ambrosia Beetle Fungi, but Inhibits Their Pathogens and Fungal Symbionts of Bark Beetles.
Bark beetles (sensu lato) colonize woody tissues like phloem or xylem and are associated with a broad range of micro-organisms. Specific fungi in the ascomycete orders Hypocreales, Microascales and Ophistomatales as well as the basidiomycete Russulales have been found to be of high importance for successful tree colonization and reproduction in many species. While fungal mutualisms are facultative for most phloem-colonizing bark beetles (sensu stricto), xylem-colonizing ambrosia beetles are long known to obligatorily depend on mutualistic fungi for nutrition of adults and larvae. Recently, a defensive role of fungal mutualists for their ambrosia beetle hosts was revealed: Few tested mutualists outcompeted other beetle-antagonistic fungi by their ability to produce, detoxify and metabolize ethanol, which is naturally occurring in stressed and/or dying trees that many ambrosia beetle species preferentially colonize. Here, we aim to test (i) how widespread beneficial effects of ethanol are among the independently evolved lineages of ambrosia beetle fungal mutualists and (ii) whether it is also present in common fungal symbionts of two bark beetle species (Ips typographus, Dendroctonus ponderosae) and some general fungal antagonists of bark and ambrosia beetle species. The majority of mutualistic ambrosia beetle fungi tested benefited (or at least were not harmed) by the presence of ethanol in terms of growth parameters (e.g., biomass), whereas fungal antagonists were inhibited. This confirms the competitive advantage of nutritional mutualists in the beetle's preferred, ethanol-containing host material. Even though most bark beetle fungi are found in the same phylogenetic lineages and ancestral to the ambrosia beetle (sensu stricto) fungi, most of them were highly negatively affected by ethanol and only a nutritional mutualist of Dendroctonus ponderosae benefited, however. This suggests that ethanol tolerance is a derived trait in nutritional fungal mutualists, particularly in ambrosia beetles that show cooperative farming of their fungi.


[Intracoronary administration of ergonovine maleate for detecting vasospastic angina; one dose method].
Coronary spasm is an important etiologic mechanism in the pathogenesis of myocardial ischemia. Provocative test of coronary spasm during coronary arteriography is clinically useful. The ergonovine test has gained widespread use, and we have examined the efficacy and safety of intracoronary ergonovine application with a fixed dose of 16 micrograms. We studied 119 patients undergoing coronary arteriography. Coronary spasm was induced in 34 cases by intracoronary administration of 16 micrograms of ergonovine maleate. Coronary spasm was readily resolved by intracoronary administration of isosorbide dinitrate. None of the cases negative to the intracoronary ergonovine applications could be induced by additional systemic administration of 0.4 mg of ergonovine. Side effects of ergonovine such as elevation of blood pressure, headache and chest symptoms were infrequent in the intracoronary ergonovine test. We conclude that our method of intracoronary ergonovine application is sensitive and safe for the diagnosis of coronary spasm.


Effects of intensive low-salt diet education on albuminuria among nondiabetic patients with hypertension treated with olmesartan: a single-blinded randomized, controlled trial.
The antiproteinuric effect of a renin-angiotensin-aldosterone system blockade can be magnified by dietary salt restriction. This study sought to determine the effect of intensive low-salt diet education on BP and urine albumin excretion in nondiabetic patients with hypertension and albuminuria. This study was conducted between March of 2012 and March of 2013 as an open-label, randomized, controlled trial. After a run-in period of 8 weeks, all patients received the angiotensin II receptor blocker olmesartan (40 mg daily). Patients were then divided into two groups. One group was treated for another 8 weeks with angiotensin II receptor blocker plus conventional low-salt diet education, and the other group was treated for 8 weeks with angiotensin II receptor blocker plus intensive low-salt diet education. The final analyses was performed with 245 completed patients. The amount of daily albuminuria was significantly decreased from 0 (566.0 [25.0-5398.6] mg/d) to 8 weeks (282.5 [16.1-4898.5] mg/d; P<0.001). From 8 to 16 weeks, the 24-hour urinary sodium excretion was decreased by 36.0±5.9 mmol/d in the intensive education group and 8.8±4.9 mmol/d in the conventional education group (interaction P<0.001). Patients who completed intensive low-salt diet education exhibited greater decreases in urinary albumin excretion than the control group (change in albuminuria from 8 to 16 weeks, -154.0 versus 0.4 mg/d; P=0.01). Urinary albumin excretion tended to decrease as the 24-hour urinary sodium excretion amount decreased (R=0.32; 95% confidence interval, 0.20 to 0.43; P<0.001). The 24-hour urinary albumin excretion was decreased more in patients in the intensive low-salt diet education group than patients in the conventional education group. Weekly intensive education on a low-salt diet would be a suitable method for clinical practice.


Immediate effects of lumbar spine manipulation on the resting and contraction thickness of transversus abdominis in asymptomatic individuals.
Randomized, blinded, controlled crossover trial. To determine if thrust joint manipulation (TJM) to the lumbar spine would result in changes to the resting and contraction thickness of transversus abdominis (TrA) in healthy individuals. Recent studies have demonstrated an immediate decrease in resting thickness and an increase in contraction thickness in TrA following lumbar TJM in patients with low back pain (LBP) who met a clinical prediction rule (CPR) for spinal manipulation. This observed phenomenon has not been investigated in healthy individuals. Thirty-five healthy participants were randomly assigned to receive a TJM or sham manipulation treatment. All participants received instruction on how to produce an isolated concentric contraction of the TrA that involved visual ultrasound imaging biofeedback. Data were analyzed using ultrasound imaging to measure changes in thickness of the TrA at rest and during contraction, following the administration of each treatment. There were no interactions observed between treatment and time for TrA muscle thickness at rest (P = .351) and during the contracted state (P = .761). Our results indicate that TJM to the lumbar spine does not appear to affect the resting or contraction thickness of TrA in healthy individuals. These findings are in contrast to previous research in which patients with LBP who met a CPR demonstrated an immediate decrease in resting thickness and an increase in contraction thickness in TrA following lumbar TJM.


Prevalence and risk factors for diabetic retinopathy: the Beijing Communities Diabetes Study 6.
To examine prevalence and associated factors of diabetic retinopathy in patients with Type 2 diabetes mellitus in urban communities of Beijing. The community health care center-based study included subjects with diabetes mellitus and an age of 20 years to 80 years, who were recruited from 15 community health centers in urban Beijing. Diabetes mellitus was defined using the World Health Organization criteria. Fundus photographs were graded using the modified Airlie House classification system. Of 2,642 eligible patients, 2,007 (76.0%) subjects (1,199 women) with a mean age of 64.1 ± 9.0 years participated. The overall prevalence of diabetic retinopathy was 24.7 ± 1.0% (95% confidence interval [CI], 22.8-26.6). In binary logistic analysis, presence of diabetic retinopathy was associated with younger age (odds ratio [OR], 0.97; 95% CI, 0.95-0.98), longer duration of diabetes (OR, 1.10; 95% CI, 1.08-1.12), higher concentration of glycosylated hemoglobin HbA1c (OR, 1.23; 95% CI, 1.14-1.33), higher systolic blood pressure (OR, 1.01; 95% CI, 1.01-1.02), lower body mass index (OR, 0.95; 95% CI, 0.92-0.98), and elevated blood urea concentration (OR, 1.01; 95% CI, 1.00-1.01). Microalbuminuria was an additional associated factor (OR, 1.55; 95% CI, 1.16-2.08). Patients with microalbuminuria were 4.7 times more likely to have a severe or proliferating diabetic retinopathy than those without microalbuminuria. In the urban population of Beijing, prevalence of diabetic retinopathy in diabetic patients was 25%. As in whites, increased blood pressure besides elevated plasma glucose concentrations was highly significantly associated with diabetic retinopathy in Chinese. It suggests that in Chinese as also in whites, blood pressure control beside control of plasma glucose levels is important to prevent development or progression of diabetic retinopathy.


DNA ligation catalyzed by human topoisomerase II alpha.
The DNA ligation reaction of topoisomerase II is essential for genomic integrity. However, it has been impossible to examine many fundamental aspects of this reaction because ligation assays historically required the enzyme to cleave a DNA substrate before sealing the nucleic acid break. Recently, a cleavage-independent DNA ligation assay was developed for human topoisomerase IIalpha [Bromberg, K. D., Hendricks, C., Burgin, A. B., and Osheroff, N. (2002) J. Biol. Chem. 277, 31201-31206]. This assay overcomes the requirement for DNA cleavage by monitoring the ability of the enzyme to ligate a nicked oligonucleotide in which the 5'-terminal phosphate at the nick has been activated by covalent attachment to the tyrosine mimic, p-nitrophenol. The cleavage-independent ligation assay was used to more fully characterize the DNA ligation activity of human topoisomerase IIalpha. Results suggest that the active site tyrosine contributes little to the catalysis of DNA ligation beyond its primary role as an activating/leaving group. Although arginine 804 (the residue immediately N-terminal to the active site tyrosine) has been proposed to help anchor the 5'-DNA terminus during cleavage, conversion of this residue to alanine had only a modest effect on DNA ligation. Thus, it appears that arginine 804 does not play an essential role in DNA strand joining. In contrast, disruption of base pairing at the 5'-DNA terminus abrogated DNA ligation in the absence of a covalent enzyme-DNA bond. Therefore, it is proposed that base pairing represents a secondary mechanism for aligning the 5'-DNA termini for ligation. Finally, the human enzyme appears to ligate the two scissile bonds of a cleavage site in a nonconcerted fashion.


Transition States in Single Case Experimental Designs.
The continuation of a baseline pattern of responding into a treatment phase, sometimes referred to as a "transition state," can obscure interpretation of data depicted in single-case experimental designs (SCEDs). For example, when using visual analysis, transition states may lead to the conclusion that the treatment is ineffective. Likewise, the inclusion of overlapping data points in some statistical analyses may lead to conclusions that the treatment had a small effect size and give rise to publication bias. This study reviewed 20 volumes in a journal that publishes primarily SCEDs studies. We defined a transition state as a situation wherein at least the first three consecutive data points of a treatment phase or condition are within the range of the baseline phase or condition. Results indicate that transitions states (a) were present for 7.4% of graphs that met inclusion criteria and (b) occurred for a mean of 4.9 data points before leading to behavior change. We discuss some implications and directions for future research on transition states.


Preparation and evaluation of palmitic acid-conjugated exendin-4 with delayed absorption and prolonged circulation for longer hypoglycemia.
Albumin-binding achieved by fatty-acylation to drugs is considered to be an effective means of prolonging the circulation lifetimes of short-lived peptides. Here, exendin-4 was modified with palmitic acid, and the particle size and albumin-binding of palmitic acid-conjugated exendin-4 (Pal-Ex4) purified was investigated and visualized. Additionally, its pharmacokinetics and pharmacodynamics were evaluated in diabetic rodents. Pal-Ex4 had a greater molecular size (~125nm) and its albumin-binding was 5.6-fold that of Ex4. Molecular imaging showed that the subcutaneous absorption of Pal-Ex4 was delayed until 24h post-injection, whereas Ex4 was rapidly absorbed and distributed systemically. Pharmacokinetic and pharmacodynamic results confirmed these observations, for example, times to reach peak concentration and to achieve a blood glucose level nadir were greatly delayed versus Ex4, and the circulating half-life of Pal-Ex4 was much greater than that of Ex4. Consequently, the hypoglycemic degree of Pal-Ex4 (500 nmol/kg) was 4.2 fold greater than Ex4. Our results show that the extended hypoglycemic efficacy of Pal-Ex4 was due to (i) a delayed absorption due to micelle formation and (ii) an increased in vivo circulating half-life due to albumin-binding. We believe that this prototype of exendin-4 has considerable pharmaceutical potential as a systemic type 2 anti-diabetic treatment.


A Case Report of Loperamide-Induced Ventricular Storm.
Loperamide is an easily accessible antidiarrheal medication. Unlike other medications in its class, loperamide is unique in that it causes euphoria at supratherapeutic levels due to its effect on opioid receptors. Unfortunately, with its growing abuse potential also comes increasing reports of cardiotoxicity including prolonged QT, torsades de pointes, and sudden cardiac death. We report a case of a 29-year-old female who presented with unstable arrhythmia that further progressed into electrical storm in the setting of loperamide toxicity. Due to its growing popularity and availability, it is important for clinicians to understand loperamide's mechanisms for causing toxicity as well as how to appropriately treat its complications.


Co-treatment with basic fibroblast growth factor and 17beta-estradiol in the presence of dexamethasone accelerates bone formation by rat bone marrow stromal cell culture.
Bone marrow stromal cells (BMSCs) are a promising cell source in applications for tissue engineering and regenerative medicine. Optimization and control of the growth and differentiation of cultivated cells can be achieved by the administration of growth factors and hormones in vitro. This study provided experimental information on the enhancement of the osteogenic potential of rat BMSCs in vitro and in vivo. Mineralized nodule formation of rat BMSCs in culture for 3 weeks with dexamethasone (Dex)-treated media supplemented with both basic fibroblast growth factor (bFGF) and 17beta -estradiol (E2) was examined by histology. In porous beta-tricalcium phosphate (beta - TCP), proliferation, migration, and differentiation of BMSCs were examined by histology and transmission electron microscopy. After culturing, the composites were subcutaneously implanted into syngeneic rats. The tissues with implants were harvested after 4 weeks and evaluated microscopically by using histological stain. Dex-treated media supplemented with both bFGF and E2 was the most effective in mineralized nodule formation of BMSCs in vitro. Light and electron microscopy revealed the presence of many cells with developed rough endoplasmic reticulum. Bone formation in the BMSC/beta -TCP composites in cultures in vitro for 3 weeks was observed histologically at 4 weeks after implantation. When BMSC/beta -TCP composites were cultured in Dex-treated media supplemented with both bFGF and E2, the amount of bone formation at implants was substantially greater than that of composites cultured in Dex-treated media supplemented with bFGF. The combined use of bFGF and E2 could effectively improve the bone-forming ability of BMSCs.


On the inotropic actions of arginine vasopressin in ventricular muscle of the guinea pig heart.
Arginine vasopressin (AVP) (0.03-3 microM) produced transient negative responses that were followed by slowly developing positive inotropic responses in guinea pig papillary muscles. The inotropic responses were antagonized by OPC-21268 (10 microM), but not by OPC-31260, and suppressed by neomycin (3 mM). The negative inotropic response was hardly affected by staurosporine (10 nM) and H-7 (10 microM). AVP (1 microM) elicited a transient decrease followed by a slowly developing increase in I(Ca). The latter increase in I(Ca) was not elicited in the presence of staurosporine (1 nM). These results indicate that AVP stimulates V1 receptors that couple to phosphoinositide hydrolysis and thereby increases intracellular Ca2+ and activates protein kinase C.


Management of concussion and post-concussion syndrome.
Concussion and mild traumatic brain injury (mTBI) are common clinical problems. However, the literature is not consistent in defining how concussion and mTBI are related. Although most patients with concussion recover within days to weeks, approximately 10% develop persistent signs and symptoms of post-concussion syndrome (PCS). There are no scientifically established treatments for concussion or PCS and thus rest and cognitive rehabilitation are traditionally applied, with limited effectiveness. This article presents a clinical model to suggest that concussion evolves to become mTBI after PCS has developed, representing a more severe form of brain injury. The basic pathophysiology of concussion is presented, followed by a recommended approach to the clinical evaluation of concussion in the emergency department and the physician's office. We evaluate the limited evidence-based pharmacologic treatment of acute concussion symptoms and PCS symptoms and also discuss return to activity recommendations, with an emphasis on athletes. Lastly, we suggest a promising new direction for helping patients recover from PCS.


[Specific features in pharmacokinetics of the original neuroleptic dilept in animals and humans].
Interspecies differences in pharmacokinetics of the original neuroleptic drug dilept have been studied in experimental animals (rabbits and rats) and volunteers after single oral administration of tablets and tablet mass of the drug. Parent drug in the rabbit blood plasma was detected for 4 h, in the rat plasma for about 2 h, and in the human blood plasma for about 1 h after drug administration. The degrees of dilept biotransformation into metabolites (defined as metabolism intensity, AUCM/AUCP) in rats were 21.3 (for M-1) and 1645 (for M-2), in human volunteers - 5.8 and 658.5, and in rabbits - 1.6 and 125.8, respectively. Thus, the intensity of drug metabolism in experimental animals and volunteers was different and decreased in the series rats humans rabbits.


North American opossum Didelphis virginiana as a fetal nephrotoxicity model: histologic and ultrastructural assessment of uranyl nitrate (UN)-induced damage.
We have used our opossum model of fetal nephrotoxicity to investigate uranyl nitrate-induced morphologic changes in the developing kidney. The present study establishes a renal dose response curve for the uranyl nitrate (UN). We find that pups treated with nonlethal doses of UN do not demonstrate growth retardation compared to saline-treated controls. The kidneys of UN-treated pups are heavier than the control animals, an effect less apparent the longer the pups are followed. A low dose of 60 mg/kg of UN administered to small pups causes slight histologic derangement but nevertheless more change than the same dose administered to larger more mature pups. Using a dose of 100 mg/kg of UN that effectively causes nonfatal renal disruption, we examined the kidneys from 4 to 42 days following injection. We find that tubular dilation and epithelial necrosis starts soon after treatment (day 4) and reaches its maximum during the second and third week (11 and 22 days). Architectural restoration appears complete by the end of the third week. By electron microscopy, UN induces sequential structural damage with loss of proximal tubule brush border, epithelial necrosis with intact basement membranes and regeneration at 4, 11, and 22 days. Residual tubular mitochondrial damage is present at 42 days in spite of histologically normal tubules. No apparent lesions are seen in glomeruli. Fibroblastic interstitial proliferation in UN-treated kidneys at 11 days is not followed by appreciable fibrosis when assessed at 22 and 42 days. As the structural changes caused by 100 mg/ml UN administration in fetal opossum kidneys are reversible, this is a useful model to study the molecular mediators responsible for this form of renal damage and repair.


Third-degree full-thickness burns as a complication of cervical radiofrequency ablation.
A 46-year-old woman underwent a cervical radiofrequency ablation (RFA) for chronic neck pain. Following the procedure, two areas surrounding the grounding pad in the lumbar region developed full thickness third-degree burns. Burn injuries following cervical RFA are rarely reported and are most often associated with cardiac and solid tumour RFA. Only one other case has been reported in literature with a similar outcome following a thoracic facet RFA. In our case, the lesion was directly from the ground pad and not from the radiofrequency electrode, which is more often the culprit. This is the first case reported in the literature of a full-thickness skin burn from a cervical RFA. Physicians should be aware of the potential for severe burns around the RF probe and ground pad as sequelae of RFA, and we caution the use of sedation during the procedure, as patients will unlikely be able to report any unusual sensation.


Deep Shape Features for Predicting Future Intracranial Aneurysm Growth.
Introduction: Intracranial aneurysms (IAs) are a common vascular pathology and are associated with a risk of rupture, which is often fatal. Aneurysm growth is considered a surrogate of rupture risk; therefore, the study aimed to develop and evaluate prediction models of future artificial intelligence (AI) growth based on baseline aneurysm morphology as a computer-aided treatment decision support. Materials and methods: Follow-up CT angiography (CTA) and magnetic resonance angiography (MRA) angiograms of 39 patients with 44 IAs were classified by an expert as growing and stable (25/19). From the angiograms vascular surface meshes were extracted and the aneurysm shape was characterized by established morphologic features and novel deep shape features. The features corresponding to the baseline aneurysms were used to predict future aneurysm growth using univariate thresholding, multivariate random forest and multi-layer perceptron (MLP) learning, and deep shape learning based on the PointNet++ model. Results: The proposed deep shape feature learning method achieved an accuracy of 0.82 (sensitivity = 0.96, specificity = 0.63), while the multivariate learning and univariate thresholding methods were inferior with an accuracy of up to 0.68 and 0.63, respectively. Conclusion: High-performing classification of future growing IAs renders the proposed deep shape features learning approach as the key enabling tool to manage rupture risk in the "no treatment" paradigm of patient follow-up imaging.


[Dynamics of the factors of 137Cs transfer to natural and sown grasses in the long term after the accident at the Chernobyl power plant].
The regularities of changes in factors of 137Cs transfer to natural and sown grasses in the longterm period after the Chernobyl accident are described. The dynamics of changes in factors of 137Cs transfer to sown grasses over the period of 1993 to 1999 was of a non-uniform character. On average, these transfer factors (TF) tended to decrease in that period. A significant tendency for an increase in TF to sown grasses was noted for 1997-1999. The half-life periods of decrease in TF to natural grasses in the period 1993-1999 were shown to be ranged from 4.6 to 24.6 years. It was demonstrated that the differences in the TF dynamics between sown and natural grasses were related to the peculiarities of their development and application of countermeasures.


Key Regulators of Autophagosome Closure.
Autophagy is an evolutionarily conserved pathway, in which cytoplasmic components are sequestered within double-membrane vesicles called autophagosomes and then transported into lysosomes or vacuoles for degradation. Over 40 conserved autophagy-related (ATG) genes define the core machinery for the five processes of autophagy: initiation, nucleation, elongation, closure, and fusion. In this review, we focus on one of the least well-characterized events in autophagy, namely the closure of the isolation membrane/phagophore to form the sealed autophagosome. This process is tightly regulated by ESCRT machinery, ATG proteins, Rab GTPase and Rab-related proteins, SNAREs, sphingomyelin, and calcium. We summarize recent progress in the regulation of autophagosome closure and discuss the key questions remaining to be addressed.


Nutraceuticals in the Management of Cardiovascular Risk Factors: Where is the Evidence?
Cardiovascular disease continues to rise at an alarming rate, and research focuses on possible therapies to reduce the risk and slow down its progression. Several epidemiological studies have indicated that dietary modifications, such as increased consumption of fruits and vegetables play an important role in reducing cardiovascular disease risk factors. Food sources rich in antioxidants, anti-inflammatory, hypolipidemic, and hypoglycemic properties are thought to ameliorate the progression of cardiovascular disease and serve as a potential treatment mode. Many in vivo and in vitro studies using turmeric, cinnamon, mango, blueberries, red wine, chocolate, and extra virgin olive oil have demonstrated significant improvements in cholesterol profiles, toxic reactive oxygen species, inflammation, obesity, and hypertension. In this review, we summarize recent evidence on the cardioprotective effect of different food groups, outline their potential mechanisms involved in slowing down the progression of cardiovascular disease, and highlight the beneficial effects associated with increased consumption.


Single Ca(2+)-activated Cl(-) channel currents recorded from toad olfactory cilia.
Odor transduction, occurring in the chemosensory cilia of vertebrate olfactory sensory neurons, is triggered by guanosine triphosphate-coupled odor receptors and mediated by a cyclic adenosine monophosphate (cAMP) signaling cascade, where cAMP opens cationic non-selective cyclic nucleotide-gated (CNG) channels. Calcium enters through CNG gates Ca(2+)-activated Cl(-) channels, allowing a Cl(-) inward current that enhances the depolarization initiated by the CNG-dependent inward current. The anoctamin channel 2, ANO2, is considered the main Ca(2+)-activated Cl(-) channel of olfactory transduction. Although Ca(2+)-activated Cl(-) channel-dependent currents in olfactory sensory neurons were reported to be suppressed in ANO2-knockout mice, field potentials from their olfactory epithelium were only modestly diminished and their smell-dependent behavior was unaffected, suggesting the participation of additional Ca(2+)-activated Cl(-) channel types. The Bestrophin channel 2, Best2, was also detected in mouse olfactory cilia and ClCa4l, belonging to the ClCa family of Ca(2+)-activated Cl(-) channels, were found in rat cilia. Best2 knock-out mice present no electrophysiological or behavioral impairment, while the ClCa channels have not been functionally studied; therefore, the overall participation of all these channels in olfactory transduction remains unresolved. We explored the presence of detectable Ca(2+)-activated Cl(-) channels in toad olfactory cilia by recording from inside-out membrane patches excised from individual cilia and detected unitary Cl(-) current events with a pronounced Ca(2+) dependence, corresponding to 12 and 24 pS conductances, over tenfold higher than the aforementioned channels, and a approx. fivefold higher Ca(2+) affinity (K0.5 = 0.38 µM). Remarkably, we observed immunoreactivity to anti-ClCa and anti-ANO2 antibodies in the olfactory cilia, suggesting a possible cooperative function of both channel type in chemotransduction. These results are consistent with a novel olfactory cilia channel, which might play a role in odor transduction.


Paradoxical role of interleukin-33/suppressor of tumorigenicity 2 in colorectal carcinogenesis: Progress and therapeutic potential.
Colorectal cancer (CRC) is presently the second most prevalent global mortality-inducing cancer. CRC carcinogenesis is a multifactorial process involving internal genetic mutations and the external environment. In addition, non-neoplastic cell activities within tumor microenvironments for CRC development have been established. However, interleukin (IL)-33, secreted by such cell types, plays a pivotal role in cancer progression due to interaction with cellular constituents within the tumor-inflammation microenvironment. IL-33 belongs to the IL-1 cytokine family and acts as binding attachments for the suppressor of tumorigenicity (ST)2 receptor. Therefore, how to coordinate tumor microenvironment, design and optimize treatment strategies suitable for CRC, based on IL-33/ST2 signal is a challenge. Even though it has established influences upon immunity-linked conditions, IL-33 effects on CRC progression and prevention and related mechanisms are still controversial. Our review depicts controversial activities for IL-33/ST2 within carcinogenesis and cancer prevention. Moreover, IL-33/ST2 signaling is a potential therapeutic target for CRC.


Thermochemistry of halomethanes CF(n)Br(4-n) (n = 0-3) based on iPEPICO experiments and quantum chemical computations.
Internal energy selected bromofluoromethane cations were prepared and their internal energy dependent fragmentation pathways were recorded by imaging photoelectron photoion coincidence spectroscopy (iPEPICO). The first dissociation reaction is bromine atom loss, which is followed by fluorine atom loss in CF(3)Br and CF(2)Br(2) at higher energies. Accurate 0 K appearance energies have been obtained for these processes, which are complemented by ab initio isodesmic reaction energy calculations. A thermochemical network is set up to obtain updated heats of formation of the samples and their dissociative photoionization products. Several computational methods have been benchmarked against the well-known interhalogen heats of formation. As a corollary, we stumbled upon an assignment issue for the ClF heat of formation leading to a 5.7 kJ mol(-1) error, resolved some time ago, but still lacking closure because of outdated compilations. Our CF(3)(+) appearance energy from CF(3)Br confirms the measurements of Asher and Ruscic (J. Chem. Phys. 1997, 106, 210) and Garcia et al. (J. Phys. Chem. A 2001, 105, 8296) as opposed to the most recent result of Clay et al. (J. Phys. Chem. A 2005, 109, 1541). The ionization energy of CF(3) is determined to be 9.02-9.08 eV on the basis of a previous CF(3)-Br neutral bond energy and the CF(3) heat of formation, respectively. We also show that the breakdown diagram of CFBr(3)(+), a weakly bound parent ion, can be used to obtain the accurate adiabatic ionization energy of the neutral of 10.625 ± 0.010 eV. The updated 298 K enthalpies of formation Δ(f)H(o)(g) for CF(3)Br, CF(2)Br(2), CFBr(3), and CBr(4) are reported to be -647.0 ± 3.5, -361.0 ± 7.4, -111.6 ± 7.7, and 113.7 ± 4 kJ mol(-1), respectively.


[Neurological anthropology among the Kamayura Indians of the Alto Xingu].
The Kamayura tribe is made up of 300 persons living in the Alto Xingu in Mato Grosso, Brazil. Their traditional system of health care is based on the pajé, a witch doctor who uses plants and prayers for treatment. Field work was done in the Kamayura village holding successive interviews with the chief and the pajé Takumá to obtain information regarding the neurological disorders found there, the indigenous beliefs regarding illness (natural or based on witch-craft), their classification and traditional treatment based on plants. The terms employed were translated from Kamayura into Portuguese. Illness in Indians is caused by the revenge of the spirit (mama'e) of the animal killed by the huntsman. Epilepsy (Teawurup) or armadillo disease is caused when a huntsman kills an armadillo. It is treated with two roots, tsimó and wewuru, kneaded, diluted in water and applied to the eyes. An infusion of enamum root is also used. Migraine or monkey disease causes a pulsatile headache and vertigo. Mental retardation and infantile cerebral palsy are included in the ant-bear disease. Depression is treated with the plant 'iputunú'; which is applied diluted in water to the face of the patient so that he no longer sees his dead relations and may be cured. Schizophrenia or apuayat (owl disease) also occurs, but not parkinsonism or stroke. The Kamayura pajes have established a system of health-care based on magic folklore, transmitted orally and making use of traditional plants.


Behavioral health service use post-jail release and reduced risk of return to jail.
This study examined whether behavioral health service use post-jail release was associated with reduced risk of jail reincarceration. The study sample included 20,615 individuals who had behavioral health diagnoses and were released from the Philadelphia County jail. Using administrative records of the county jail and state-, county-, and Medicaid-funded behavioral health service use from 2010 to 2018, we conducted Cox proportional hazard analyses to estimate the association between behavioral health service use post-jail release and the risk of return to jail within 3 years. Nearly 50% of the sample returned to jail within 3 years. Individuals who used behavioral health services were 26%-38% less likely to return to jail within 3 years than were individuals who did not. The study results suggest that connecting individuals with behavioral health services upon release from jail can reduce the risk of repeated jail incarceration.


The thioredoxin antioxidant system.
The thioredoxin (Trx) system, which is composed of NADPH, thioredoxin reductase (TrxR), and thioredoxin, is a key antioxidant system in defense against oxidative stress through its disulfide reductase activity regulating protein dithiol/disulfide balance. The Trx system provides the electrons to thiol-dependent peroxidases (peroxiredoxins) to remove reactive oxygen and nitrogen species with a fast reaction rate. Trx antioxidant functions are also shown by involvement in DNA and protein repair by reducing ribonucleotide reductase, methionine sulfoxide reductases, and regulating the activity of many redox-sensitive transcription factors. Moreover, Trx systems play critical roles in the immune response, virus infection, and cell death via interaction with thioredoxin-interacting protein. In mammalian cells, the cytosolic and mitochondrial Trx systems, in which TrxRs are high molecular weight selenoenzymes, together with the glutathione-glutaredoxin (Grx) system (NADPH, glutathione reductase, GSH, and Grx) control the cellular redox environment. Recently mammalian thioredoxin and glutathione systems have been found to be able to provide the electrons crossly and to serve as a backup system for each other. In contrast, bacteria TrxRs are low molecular weight enzymes with a structure and reaction mechanism distinct from mammalian TrxR. Many bacterial species possess specific thiol-dependent antioxidant systems, and the significance of the Trx system in the defense against oxidative stress is different. Particularly, the absence of a GSH-Grx system in some pathogenic bacteria such as Helicobacter pylori, Mycobacterium tuberculosis, and Staphylococcus aureus makes the bacterial Trx system essential for survival under oxidative stress. This provides an opportunity to kill these bacteria by targeting the TrxR-Trx system.


Role of microtubules in Piezo2 mechanotransduction of mouse Merkel cells.
Piezo2 channels are expressed in Merkel cells and somatosensory neurons to mediate mechanotransduction leading to the sense of touch. Components of the cytoskeleton including microtubules are key intracellular structures that maintain cellular membrane mechanics and thereby may be important in mechanotransduction. In the present study, we have explored, with microtubule-targeting agents, the potential role of microtubules in Piezo2-mediated mechanotransduction in Merkel cells of mouse whisker hair follicles. Applying patch-clamp recordings to Merkel cells in situ in whisker hair follicles, we show that Piezo2-mediated mechanically activated (MA) currents in Merkel cells are significantly potentiated by the microtubule stabilizer paclitaxel but reduced by the microtubule destabilizer vincristine. Furthermore, electrophysiological recordings made from whisker hair follicle afferent nerves show that mechanically evoked whisker afferent impulses are significantly enhanced by paclitaxel and its analog docetaxel but significantly suppressed by vincristine and its analog vinblastine. Our findings suggest that microtubules play an essential role in Piezo2 mechanotransduction in Merkel cells.NEW & NOTEWORTHY Piezo2 channels are expressed in Merkel cells to mediate mechanotransduction leading to the sense of touch. Here we determined the role of microtubules in regulating Piezo2-mediated mechanotransduction in Merkel cells. Piezo2-mediated currents in Merkel cells are potentiated by microtubule stabilizer paclitaxel but reduced by microtubule destabilizer vincristine. Mechanically evoked afferent impulses are also enhanced by microtubule stabilizers and suppressed by microtubule destabilizers. Microtubules may play an essential role in Piezo2 mechanotransduction in Merkel cells.


Methodologic Considerations in the Application of Next-Generation Sequencing of Human TRB Repertoires for Clinical Use.
Next-generation sequencing (NGS) of immune receptors has become a standard tool to assess minimal residual disease (MRD) in patients treated for lymphoid malignancy, and it is being used to study the T-cell repertoire in many clinical settings. To better understanding the potential clinical utility and limitations of this application outside of MRD, we developed a BIOMED-2 primer-based NGS method and characterized its performance in controls and patients with graft-versus-host disease (GVHD) after allogeneic hematopoietic transplant. For controls and patients with GVHD, replicate sequencing of the same T-cell receptor β (TRB) libraries was highly reproducible. Higher variability was observed in sequencing of different TRB libraries made from the same DNA stock. Variability was increased in patients with GVHD compared with controls; patients with GVHD also had lower diversity than controls. In the T-cell repertoire of a healthy person, approximately 99.6% of the CDR3 clones were in low abundance, with frequency <10-3. A single library could identify >93% of the clones with frequency ≥10-3 in the repertoire. Sequencing in duplicate increased the average detection rate to >97%. This work demonstrates that NGS reliably and robustly characterizes TRB populations in healthy individuals and patients with GVHD with frequency ≥10-3 and provides a methodologic framework for applying NGS immune repertoire methods to clinical testing applications beyond MRD.


Specific bacterial suppressors of MAMP signaling upstream of MAPKKK in Arabidopsis innate immunity.
Plants and animals possess innate immune systems to prevent infections and are effectively "nonhosts" for most potential pathogens. The molecular mechanisms underlying nonhost immunity in plants remain obscure. In Arabidopsis, nonhost/nonpathogenic Pseudomonas syringae sustains but pathogenic P. syringae suppresses early MAMP (microbe-associated molecular pattern) marker-gene activation. We performed a cell-based genetic screen of virulence factors and identified AvrPto and AvrPtoB as potent and unique suppressors of early-defense gene transcription and MAP kinase (MAPK) signaling. Unlike effectors of mammalian pathogens, AvrPto and AvrPtoB intercept multiple MAMP-mediated signaling upstream of MAPKKK at the plasma membrane linked to the receptor. In transgenic Arabidopsis, AvrPto blocks early MAMP signaling and enables nonhost P. syringae growth. Deletions of avrPto and avrPtoB from pathogenic P. syringae reduce its virulence. The studies reveal a fundamental role of MAMP signaling in nonhost immunity, and a novel action of type III effectors from pathogenic bacteria.


False positive test results for pheochromocytoma from 2000 to 2008.
Testing for pheochromocytoma becomes more frequent in clinical practice. False positive test results may cause patient anxiety and unnecessary imaging studies. The data on false positive results for pheochromocytoma in routine clinical practice are lacking. To examine the prevalence of false positive results and to reveal the clinical features and laboratory tests of patients with markedly elevated but false positive test results, a database of tests for pheochromocytoma at a large general hospital between 2000 and 2008 was reviewed. Of 1,896 patients tested, 417 (22.0%) had at least one abnormal test result and 66 (3.5%) had markedly elevated results. 24 patients with markedly elevated but false positive results received 65 imaging studies and 1 adrenalectomy. The causes of the misleading results included physiological variations (33%), laboratory errors (29%), and drug interference with measurement (21%). The false positive rate of a markedly elevated result was lowest for vanillylmandelic acid (9%) and highest for urine metanephrines (50%) (p = 0.03). Nearly half of all test results were normal and 79% of patients had at least one normal result. Therefore false positive test results for pheochromocytoma are rather common. Markedly elevated but false positive test results can potentially be avoided by judicious selection of patients and tests. Pretest risk, physiological variations of catecholamine levels, laboratory errors, and drug interference should be considered in interpreting abnormal test results.


Contribution to the mode of action of univalent gold.
Au+ binds to the collagen structure and, therefore, the authors studied the kinetics of the interaction between sodium gold thiosulphate (SGTS) and C1q complement subcomponent, the structure of is partially collagenous. The kinetics was evaluated densitometrically and compared with that of collagen and SGTS. On the basis of those results, the effect of SGTS upon collagen type-II-induced arthritis (CIA) was investigated in Wistar rats. SGTS (20 mg per kg of body weight weekly) was administered i.m. starting i) with the first immunization dose, ii) with the second immunization dose, and iii) after the onset of arthritis. Regardless of the timing of drug administration the manifestation of arthritis was decreased, but the decrease was more expressed in the first two groups. It was concluded that SGTS administration is capable of inhibiting CIA, provided that the drug is applied sufficiently early. Since formation of antibodies to type-II collagen remained unaffected, it is feasible that the mode of action of gold complexes is based mainly on blocking the activation of complement system.


[Differential diagnosis and medical treatment in Cushing's disease].
Cushing's disease remains a difficult diagnosis in spite of new technical procedures such as pituitary MRI, selective bilateral petrosal or cavernous sampling, (111)In pentreotide scan and 18 Flurodeoxyglucose pituitary PET scan. In this article, we review biological diagnostic procedures of Cushing's disease and corticotroph adenomas. According to our experience and the literature, we summarize the approach in medical treatment of Cushing's disease.


In vitro evaluation of powders for inhalation: the effect of drug concentration on particle detachment.
Limited information on the effect of the drug concentration on the performance of powders for inhalation is currently published. The aim of this work was to study the influence of drug concentration on the adhesion between drug and carrier and on the drug detachment from the carrier. The study was done with formoterol fumarate and fluticasone propionate blended with lactose Lactohale 200. To assess the adhesion of respirable-sized drug to carrier particles, a simple method was developed based on aspiration and considering the whole blend as it is used in dry powder inhalers. Adhesion characteristics were evaluated by submitting the mixtures to a sieving action by air depression with an Alpine air-jet sieve. Aerodynamic evaluation of fine particle dose and emitted dose was obtained using a Twin Stage Impinger (TSI). Drug concentration of powder blends used in dry powder inhalers influenced adhesion, content uniformity and in vitro deposition of the drug. For the higher concentration of formoterol, it seemed that a lower quantity of drug adhered to the lactose. This was confirmed by the aerosolization assays done in the TSI. The fine particle fraction increased linearly with the formoterol concentration. A correlation was observed between adhesion characteristics and inertial impaction. In the case of fluticasone, the influence of the concentration was different. First, the fine particle fraction increased with the concentration and then decreased with a further increase of the fluticasone concentration. This could be explained by the lack of homogeneity when the fluticasone concentration was high because of agglomerates of pure drug which can not be redispersed, or by the physico-chemical characteristics of this drug.


Investigation into the mechanism and dynamics of DNA association and dissociation utilizing kinetic Monte Carlo simulations.
In this work, we present a kinetic Markov state Monte Carlo model designed to complement temperature-jump (T-jump) infrared spectroscopy experiments probing the kinetics and dynamics of short DNA oligonucleotides. The model is designed to be accessible to experimental researchers in terms of both computational simplicity and expense while providing detailed insights beyond those provided by experimental methods. The model is an extension of a thermodynamic lattice model for DNA hybridization utilizing the formalism of the nucleation-zipper mechanism. Association and dissociation trajectories were generated utilizing the Gillespie algorithm and parameters determined via fitting the association and dissociation timescales to previously published experimental data. Terminal end fraying, experimentally observed following a rapid T-jump, in the sequence 5'-ATATGCATAT-3' was replicated by the model that also demonstrated that experimentally observed fast dynamics in the sequences 5'-C(AT)nG-3', where n = 2-6, were also due to terminal end fraying. The dominant association pathways, isolated by transition pathway theory, showed two primary motifs: initiating at or next to a G:C base pair, which is enthalpically favorable and related to the increased strength of G:C base pairs, and initiating in the center of the sequence, which is entropically favorable and related to minimizing the penalty associated with the decrease in configurational entropy due to hybridization.


Impact of Ions on Film Conformality and Crystallinity during Plasma-Assisted Atomic Layer Deposition of TiO2.
This work demonstrates that ions have a strong impact on the growth per cycle (GPC) and material properties during plasma-assisted atomic layer deposition (ALD) of TiO2 (titanium dioxide), even under mild plasma conditions with low-energy (<20 eV) ions. Using vertical trench nanostructures and microscopic cavity structures that locally block the flux of ions, it is observed that the impact of (low-energy) ions is an important factor for the TiO2 film conformality. Specifically, it is demonstrated that the GPC in terms of film thickness can increase by 20 to >200% under the influence of ions, which is correlated with an increase in film crystallinity and an associated strong reduction in the wet etch rate (in 30:1 buffered HF). The magnitude of the influence of ions is observed to depend on multiple parameters such as the deposition temperature, plasma exposure time, and ion energy, which may all be used to minimize or exploit this effect. For example, a relatively moderate influence of ions is observed at 200 °C when using short plasma steps and a grounded substrate, providing a low ion-energy dose of ∼1 eV nm-2 cycle-1, while a high effect is obtained when using extended plasma exposures or substrate biasing (∼100 eV nm-2 cycle-1). This work on TiO2 shows that detailed insight into the role of ions during plasma ALD is essential for precisely controlling the film conformality, material properties, and process reproducibility.


Zinc supplementation as an adjunct to antibiotics in the treatment of pneumonia in children 2 to 59 months of age.
Diarrhoeal disorders and acute respiratory infections (ARIs), especially pneumonia, are the most common causes of death in low-income countries. Studies evaluating the impact of zinc supplementation as an adjunct in the management of pneumonia are limited and have shown variable results. To evaluate zinc supplementation, as an adjunct to antibiotics, in the treatment (clinical recovery) of pneumonia in children aged two to 59 months. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2011, Issue 1), which contains the Cochrane Acute Respiratory Infections (ARI) Group's and the Cochrane Infectious Diseases Group's Specialised Registers, MEDLINE (1950 to March week 2, 2011), EMBASE (1974 to March 2011), CINAHL (1981 to March 2011), LILACS (1985 to March 2011), AMED (1985 to March 2011), CAB Abstracts (1910 to March 2011) and Web of Science (2000 to March 2011). Randomised control trials (RCTs) evaluating supplementation of zinc as an adjunct to antibiotics for pneumonia in children aged two to 59 months. Two review authors independently assessed trial eligibility and screened all available titles and abstracts for inclusion. If the relevance could not be ascertained by screening the title and abstract, we retrieved and reviewed the full text of the article. We included four trials in which 3267 children aged two to 35 months participated. Analysis showed that zinc supplementation in addition to standard antibiotic therapy in children with severe and non-severe pneumonia failed to show a statistically significant effect on clinical recovery (risk ratio (RR) 1.02; 95% confidence interval (CI) 0.93 to 1.11). Similary, zinc supplementation in children with severe pneumonia, as an adjunct to standard antibiotic therapy, did not show a statistically significant effect on clinical recovery measured as resolution of tachypnoea (respiratory rate > 50 breaths per minute) (RR 1.13; 95% CI 0.82 to 1.57) and cessation of chest indrawing (RR 1.08; 95% CI 0.88 to 1.31) as compared to the control group. Zinc supplementation in children with severe pneumonia also showed a non-significant effect on the duration of hospitalization stay as compared to the control (RR 1.04; 95% CI 0.89 to 1.22). Evidence provided in this review is insufficient to recommend the use of zinc as an adjunct to standard antibiotic therapy for pneumonia in children aged two to 35 months.