The Epstein-Barr virus nuclear antigen 2 (EBNA2), a protein required for B lymphocyte immortalization, induces the synthesis of type I interferon in Burkitt's lymphoma cell lines.
Epstein-Barr virus nuclear antigen 2 (EBNA2), a protein involved in cell transformation, interferes with the cellular response to type I interferons (IFN-alpha/beta). We investigated the function of conditionally expressed EBNA2 in the context of the IFN response in Burkitt's lymphoma cell lines. Expression of EBNA2 led to the transcriptional activation of both endogenous or transfected IFN-stimulated genes (ISGs), genes which contain within their promoters either the interferon-stimulated response element (ISRE) or the gamma interferon activation site (GAS). In search of a molecular mechanism for the transcriptional induction of ISGs, we observed an EBNA2-dependent synthesis of IFN-beta mRNA at low levels and the secretion of low amounts of IFN. A transfected IFN-beta promoter responded to EBNA2 activation, and a sequence closely resembling a RBP-Jkappa binding site was pinpointed as a potential target of EBNA2 activity. EBNA2-dependent transcriptional induction of the IFN-beta promoter occurred in EBV-negative Burkitt's lymphoma cells, indicating that other EBV genes were not required for the induction of IFN-beta synthesis.


Effect of in vitro exposure to benzo[a]pyrene, a component of cigarette smoke, on folliculogenesis, steroidogenesis and oocyte nuclear maturation.
We previoulsy quantified the concentration of benzo[a]pyrene (B[a]P) in the follicular fluid of women exposed to mainstream and/or sidestream cigarette smoke. The objective of this study was to quantify the effects of B[a]P-exposure, at concentrations representative of follicluar fluid concentrations, on folliculogenesis, on gonadal steroid and anti-müllerian hormone (AMH) output, oocyte growth, and nuclear maturation. Follicles (100-130 μm) isolated from ovaries of F1 hybrid (C57BL/6j×CBA/Ca) mice were cultured for 13 days in increasing concentrations of B[a]P (0 ng/ml (control) to 45 ng/ml). B[a]P treatment inhibited (p < 0 .05) antral follicle development, decreased estradiol output and follicle survival at the 45.0 ng/ml dose. B[a]P exposure decreased AMH output overall during preantral (p = 0.014) and antral (p = 0.026) follicle development but had no effect on progesterone output or oocyte growth and nuclear maturation in surviving follicles. These data suggest that B[a]P is an important toxic component of cigarette smoke that adversely affects follicular development and survival.


Jmjd3 is essential for the epigenetic modulation of microglia phenotypes in the immune pathogenesis of Parkinson's disease.
Classical activation (M1 phenotype) and alternative activation (M2 phenotype) are the two polars of microglial activation states that can produce either detrimental or beneficial effects in the central nervous system (CNS). Harnessing the beneficial properties of microglia cells by modulating their polarization states provides great potential for the treatment of Parkinson's disease (PD). However, the epigenetic mechanism that regulates microglia polarization remains elusive. Here, we reported that histone H3K27me3 demethylase Jumonji domain containing 3 (Jmjd3) was essential for M2 microglia polarization. Suppression of Jmjd3 in N9 microglia inhibited M2 polarization and simultaneously exaggerated M1 microglial inflammatory responses, which led to extensive neuron death in vitro. We also observed that the suppression of Jmjd3 in the substantia nigra (SN) in vivo dramatically caused microglial overactivation and exacerbated dopamine (DA) neuron death in 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-intoxicated mouse model of PD. Moreover, we showed that the Jmjd3 level was lower in the midbrain of aged mice, which was accompanied by an elevated level of H3K27me3 and an increased ratio of M1 to M2 markers, suggesting that aging is an important factor in switching the microglia phenotypes. Overall, our studies indicate that Jmjd3 is able to enhance the polarization of M2 microglia by modifying histone H3K27me3, and therefore it has a pivotal role in the switch of microglia phenotypes that may contribute to the immune pathogenesis of PD.


Determining the feasibility of a heart transplant program.
The careful information gathering by the task force provided a foundation for informed decision making. Furthermore, the participation of the affected constituencies in the planning process helped in building a consensus in support of the new program. The group felt confident that the relevant risks and potential benefits had been identified in the feasibility assessment. Finally, the financial forecasts provided quantified benchmarks to use in measuring the performance of the program on ongoing basis.


Assembly of ribosomal subunits affected in a ribosomal mutant of E. coli having an altered L22 protein.
In this article we describe some in vivo properties of a coldsensitive ribosomal mutant from Escherichia coli. The mutation affects the rplV gene which is the structural gene of ribosomal protein L22. Our work shows that at 22 degrees C, the biosynthesis of both ribosomal subunits and the maturation processing of 15S and 23S ribosomal RNA are impaired. Integration of our results in a general model of in vivo ribosomal assembly in E. coli is presented.


Rate of novel host invasion affects adaptability of evolving RNA virus lineages.
Although differing rates of environmental turnover should be consequential for the dynamics of adaptive change, this idea has been rarely examined outside of theory. In particular, the importance of RNA viruses in disease emergence warrants experiments testing how differing rates of novel host invasion may impact the ability of viruses to adaptively shift onto a novel host. To test whether the rate of environmental turnover influences adaptation, we experimentally evolved 144 Sindbis virus lineages in replicated tissue-culture environments, which transitioned from being dominated by a permissive host cell type to a novel host cell type. The rate at which the novel host 'invaded' the environment varied by treatment. The fitness (growth rate) of evolved virus populations was measured on each host type, and molecular substitutions were mapped via whole genome consensus sequencing. Results showed that virus populations more consistently reached high fitness levels on the novel host when the novel host 'invaded' the environment more gradually, and gradual invasion resulted in less variable genomic outcomes. Moreover, virus populations that experienced a rapid shift onto the novel host converged upon different genotypes than populations that experienced a gradual shift onto the novel host, suggesting a strong effect of historical contingency.


EAU guidelines for the management of genitourinary tuberculosis.
Nearly one third of the world's population is estimated to be infected with Mycobacterium tuberculosis. Moreover, tuberculosis is the most common opportunistic infection in AIDS patients. Genitourinary tuberculosis is not very common but it is considered as a severe form of extra-pulmonary tuberculosis The diagnosis of genitourinary tuberculosis is made based on culture studies by isolation of the causative organism; however, biopsy material on conventional solid media may occasionally be required. Drug treatment is the first line therapy in genitourinary tuberculosis. Treatment regimens of 6 months are effective in most of the patients. Although chemotherapy is the mainstay of treatment, surgery in the form of ablation or reconstruction may be unavoidable. Both radical and reconstructive surgery should be carried out in the first 2 months of intensive chemotherapy.


Vitamin A deficiency and associated factors in children in urban areas.
To estimate the prevalence of vitamin A deficiency and its associated factors in children. A cross-sectional population-based study, involving 1,211 children of both sexes, aged between six and 59 months old, was carried out in the urban zone of 9 cities in the state of Paraiba, Northeastern Brazil. Vitamin A status was assessed by serum retinol levels (high performance liquid chromatography - HPLC) and subclinical infection was assessed by C-reactive protein concentrations. Socioeconomic, demographic and sanitation conditions, as well as vitamin A supplement intake, were also evaluated. Children with serum retinol concentrations The prevalence of vitamin A deficiency was 21.8% (95%CI 19.6;24.2), showing an association with subclinical infection and lack of indoor plumbing. The prevalence of vitamin A deficiency was 21.8% (95%CI 19.6;24.2). After adjustment, vitamin A deficiency was found to be linked with subclinical infection and lack of indoor plumbing. Vitamin A deficiency was four times higher (CI95% 1.49;10.16) in children with subclinical infection whose homes were without indoor plumbing, compared to children who were not infected and with indoor plumbing in their homes. Despite activities aimed at the prevention and control of vitamin A deficiency, hypovitaminosis A, remains a public health concern among children under five.


HIV-related stigma within communities of gay men: a literature review.
While stigma associated with HIV infection is well recognised, there is limited information on the impact of HIV-related stigma between men who have sex with men and within communities of gay men. The consequences of HIV-related stigma can be personal and community-wide, including impacts on mood and emotional well-being, prevention, testing behaviour, and mental and general health. This review of the literature reports a growing division between HIV-positive and HIV-negative gay men, and a fragmentation of gay communities based along lines of perceived or actual HIV status. The literature includes multiple references to HIV stigma and discrimination between gay men, men who have sex with men, and among and between many gay communities. This HIV stigma takes diverse forms and can incorporate aspects of social exclusion, ageism, discrimination based on physical appearance and health status, rejection and violence. By compiling the available information on this understudied form of HIV-related discrimination, we hope to better understand and target research and countermeasures aimed at reducing its impact at multiple levels.


Pharmacogenetics in inflammatory bowel disease.
Pharmacogenetics is the study of the association between variability in drug response and (or) drug toxicity and polymorphisms in genes. The goal of this field of science is to adapt drugs to a patient's specific genetic background and therefore make them more efficacious and safe. In this article we describe the variants in genes that influence either the efficacy or toxicity of common drugs used in the treatment of inflammatory bowel diseases (IBD), ulcerative colitis (UC), and Crohn's disease (CD) including sulfasalazine and mesalazine, azathioprine (AZA) and 6-mercaptopurine (6-MP), methotrexate (MTX), glucocorticosteroids (CSs) and infliximab. Furthermore, difficulties with pharmacogenetic studies in general and more specifically in IBD are described. Although pharmacogenetics is a promising field that already contributed to a better understanding of some of the underlying mechanisms of action of drugs used in IBD, the only discovery translated until now into daily practice is the relation between thiopurine S-methyltransferase (TPMT) gene polymorphisms and hematological toxicity of thiopurine treatment. In the future it is necessary to organize studies in well characterized patient cohorts who have been uniformly treated and systematically evaluated in order to quantitate drug response more objectively. An effort should be made to collect genomic DNA from all patients enrolled in clinical drug trials after appropriate informed consent for pharmacogenetic studies.


The temporal course of adenovirus-mediated gene transfer in the rat larynx.
Polypeptide growth factors have important influences on wound-healing and scar tissue formation. Specific growth factors or their inhibitors may potentially decrease scar tissue formation and prevent subglottic stenosis. Gene transfer using recombinant adenovirus may be an ideal method to mediate endogenous production of growth factors to inhibit fibrosis. The study incorporated adenovirus-mediated transduction of normal and stenotic rat larynges and histologic analysis of the sequential expression of a beta-galactosidase marker gene over time. The study was conducted at the animal care facility of an academic children's hospital. We report successful transduction in normal and injured rat larynx with peak expression of beta-galactosidase at 2 days after transduction and almost complete disappearance by 7 days. There appeared to be an early inflammatory response to the viral injection, but at 7 and 14 days after injection (transduction) the uninjured rat larynges resumed a normal histologic appearance. All distant sites stained negative for beta-galactosidase. Recombinant adenovirus-mediated gene transfer is feasible in the rat larynx with transient duration and limited toxicity. Adenovirus-mediated gene transfer has the potential to deliver growth factors that modulate wound healing and inflammation in the larynx by inhibiting fibrosis.


Complex regional pain syndrome complicating total knee arthroplasty.
To compare the long-term outcome of patients diagnosed with complex regional pain syndrome-type 1 (CRPS-1) after total knee arthroplasty (TKA) with those of uncomplicated TKA knees and preoperative osteoarthritic knees. Medical records of 1280 patients who underwent TKA for osteoarthritis were retrospectively reviewed; 8 were diagnosed as having symptoms and signs consistent with CRPS after TKA. Patients with primary inflammatory arthritis, signs of component loosening, malpositioning, or of infected arthroplasty were excluded. No patient had signs of CRPS prior to operative intervention. The 8 patients were compared with 2 groups of age- and sex-matched controls: uncomplicated TKA knees and preoperative osteoarthritic knees. Patients were followed up for a mean of 54 (range, 13-111) months and their range of movement, Western Ontario and McMaster Universities Osteoarthritis Index, SF-36 questionnaire scores, and Knee Society scores were assessed and compared. After appropriate treatment, most CRPS complicated patients had similar scores on SF-36, Western Ontario and McMaster Universities Osteoarthritis Index, and Knee Society scores when compared with uncomplicated TKA patients. Scores for CRPS complicated patients were significantly improved when compared with preoperative osteoarthritic patients. The incidence of CRPS after TKA was 0.7%. When managed early, patients complicated with CRPS after TKA have a similar prognosis to patients with uncomplicated TKA.


Na+ tolerance and Na+ transport in higher plants.
Tolerance to high soil [Na(+)] involves processes in many different parts of the plant, and is manifested in a wide range of specializations at disparate levels of organization, such as gross morphology, membrane transport, biochemistry and gene transcription. Multiple adaptations to high [Na(+)] operate concurrently within a particular plant, and mechanisms of tolerance show large taxonomic variation. These mechanisms can occur in all cells within the plant, or can occur in specific cell types, reflecting adaptations at two major levels of organization: those that confer tolerance to individual cells, and those that contribute to tolerance not of cells per se, but of the whole plant. Salt-tolerant cells can contribute to salt tolerance of plants; but we suggest that equally important in a wide range of conditions are processes involving the management of Na(+) movements within the plant. These require specific cell types in specific locations within the plant catalysing transport in a coordinated manner. For further understanding of whole plant tolerance, we require more knowledge of cell-specific transport processes and the consequences of manipulation of transporters and signalling elements in specific cell types.


[Drug therapy of sepsis. An indication for pentoxifylline?].
The interaction of immunocompetent cells with the vascular endothelium is of prime importance for the development of septic multiple organ failure. There is evidence from in-vitro studies that the methylxanthine derivative pentoxifylline can attenuate the extent of inflammatory reactions by amplification of cell-derived endogenous regulators. For instance, pentoxifylline potentiates the anti-inflammatory actions of adenosine, prostacyclin, and prostaglandins of the E series by its synergistic action on intracellular cyclic AMP. By this mechanism, pentoxifylline inhibits the oxygen-radical production of polymorphonuclear leukocytes, the aggregation of platelets, disseminated intravascular coagulation, and the production of cytokines. Consequently, pentoxifylline improves perfusion in the microcirculation as well as tissue oxygenation. Further studies will clarify whether the promising results obtained with pentoxifylline in experimental septic shock will be confirmed under clinical conditions.


'The body seems to have no life': The experiences and perceptions of fatigue among patients after COVID-19.
This study aimed at determining the perception of fatigue among patients with a history of the coronavirus disease (COVID-19). Fatigue is a long-lasting distressing symptom. It is a multidimensional symptom consisting of several factors, including physiological, psychological, social and environmental. It is vital to examine and understand the perception of fatigue among post-COVID-19 participants. A descriptive phenomenological design. The study sample consisted of 14 post-COVID-19 participants that were recruited using criterion sampling. The fatigue levels of the participants were determined using the Chalder Fatigue Scale (CFS), and those with a fatigue score above 12 were interviewed. All the interviews were conducted with a smartphone due to the COVID-19. The COnsolidated criteria for REporting Qualitative research (COREQ) checklist was used. The age of the participants varied from 24 to 67 years, with the majority of the participants being female (n = 8). The COVID-19 duration ranged from one to 11 months, and the CFS scores varied between 14 and 33. Four themes emerged following the qualitative data analysis: a new symptom beyond fatigue, fatigue increases dependency in daily life, fatigue impedes sociability and a way to hold on to life's regular rhythms. This study concluded that fatigue in post-COVID-19 participants is a new experience that is difficult to define and manage and overwhelmingly affects the physical and social aspects of life. Participants look for new ways to live with fatigue and turn to traditional methods and psychosocial strategies. This study revealed the miscellaneous aspects of fatigue in post-COVID-19 participants. Nurses should evaluate fatigue with a holistic approach that includes its physical, social, emotional and spiritual aspects. Nurses can play an active role in the management of fatigue, which is a very common symptom in the COVID-19 pandemic.


Maderas Rainforest Conservancy: A One Health approach to conservation.
Maderas Rainforest Conservancy (MRC) was incorporated as a conservation nonprofit organization in 2008, and manages two sites where biological field courses have been offered since the 1990s: La Suerte Biological Research Station in Costa Rica, and Ometepe Biological Research Station in Nicaragua. MRC employs a One Health approach to conservation education, and can serve as a model for other biological field sites. The Nicaraguan Molina family, who owns the sites, partnered with primatologist Paul Garber in 1994 to develop a primate field course aimed at introducing university students to field research. Through using their land to further conservation education and research, the Molina family has preserved the forest and engaged the local communities near their sites. Eight graduate theses and 46 refereed publications have been completed since 2010 based on research undertaken at MRC sites. While primate field courses have been offered at least once annually since 1994 and remain popular, a range of other ecological courses are now additionally offered. MRC operates from a One Health perspective, engaging in forest restoration and ecological monitoring projects, and has gradually expanded community outreach initiatives. MRC now conducts regular medical and veterinary missions in the communities surrounding the research stations which provide health care to local people and limit the population growth of domestic animals, thereby increasing the survival of wild animals. MRC is also active in ESL-teaching and conservation education, and funds Proyecto Jade, which empowers local women to make and sell organic jewelry. Through these programs, MRC works to help the local communities live more sustainably with the environment around them. MRC's support of research, commitment to education, medical and veterinary missions, and outreach initiatives to the local community all work together for the well-being of both the people and the environment, thus exemplifying the One Health perspective.


An approach for assessing human health vulnerability and public health interventions to adapt to climate change.
Assessments of the potential human health impacts of climate change are needed to inform the development of adaptation strategies, policies, and measures to lessen projected adverse impacts. We developed methods for country-level assessments to help policy makers make evidence-based decisions to increase resilience to current and future climates, and to provide information for national communications to the United Nations Framework Convention on Climate Change. The steps in an assessment should include the following: a) determine the scope of the assessment; b) describe the current distribution and burden of climate-sensitive health determinants and outcomes; c) identify and describe current strategies, policies, and measures designed to reduce the burden of climate-sensitive health determinants and outcomes; d) review the health implications of the potential impacts of climate variability and change in other sectors; e) estimate the future potential health impacts using scenarios of future changes in climate, socioeconomic, and other factors; f) synthesize the results; and g) identify additional adaptation policies and measures to reduce potential negative health impacts. Key issues for ensuring that an assessment is informative, timely, and useful include stakeholder involvement, an adequate management structure, and a communication strategy.


A cluster randomized trial comparing two interventions to improve treatment of major depression in primary care.
Many patients with major depression are non-adherent to antidepressant medication and do not receive care according to current guidelines. There is increasing evidence that treatment of depression in primary care can be improved. Comparison between effective interventions may help to establish the active ingredients of such interventions. In a randomized trial two interventions to improve treatment of major depression in primary care were compared (1) a depression care programme, targeting general practitioners (GPs), patients, and systematic follow-up, and (2) a systematic follow-up programme. Thirty GPs were randomized and 211 primary-care patients with current major depression were included. All patients were prescribed a selective serotonin reuptake inhibitor. Outcome measures included adherence to antidepressant medication, and depression outcome. No significant differences in adherence rates and treatment outcome measures were demonstrated between interventions at week 10 or week 26. Adherence rates were high and treatment outcome was favourable. The depression care programme was not superior to the systematic follow-up programme. Systematic follow-up in depression treatment in primary care seems to be an intervention per se, having the potential to improve adherence and treatment outcome.


Gut-Evolved Candida albicans Induces Metabolic Changes in Neutrophils.
Serial passaging of the human fungal pathogen Candida albicans in the gastrointestinal tract of antibiotics-treated mice selects for virulence-attenuated strains. These gut-evolved strains protect the host from infection by a wide range of pathogens via trained immunity. Here, we further investigated the molecular and cellular mechanisms underlying this innate immune memory. Both Dectin-1 (the main receptor for β-glucan; a well-described immune training molecule in the fungal cell wall) and Nod2 (a receptor described to mediate BCG-induced trained immunity), were redundant for the protection induced by gut-evolved C. albicans against a virulent C. albicans strain, suggesting that gut-evolved C. albicans strains induce trained immunity via other pathways. Cytometry by time of flight (CyTOF) analysis of mouse splenocytes revealed that immunization with gut-evolved C. albicans resulted in an expansion of neutrophils and a reduction in natural killer (NK) cells, but no significant numeric changes in monocytes, macrophages or dendritic cell populations. Systemic depletion of phagocytes or neutrophils, but not of macrophages or NK cells, reduced protection mediated by gut-evolved C. albicans. Splenocytes and bone marrow cells of mice immunized with gut-evolved C. albicans demonstrated metabolic changes. In particular, splenic neutrophils displayed significantly elevated glycolytic and respiratory activity in comparison to those from mock-immunized mice. Although further investigation is required for fully deciphering the trained immunity mechanism induced by gut-evolved C. albicans strains, this data is consistent with the existence of several mechanisms of trained immunity, triggered by different training stimuli and involving different immune molecules and cell types.


Determining current physical therapist management of hip fracture in an acute care hospital and physical therapists' rationale for this management.
Physical therapy has an important role in hip fracture rehabilitation to address issues of mobility and function, yet current best practice guidelines fail to make recommendations for specific physical therapy interventions beyond the first 24 hours postsurgery. The aims of this study were: (1) to gain an understanding of current physical therapist practice in an Australian acute care setting and (2) to determine what physical therapists consider to be best practice physical therapist management and their rationale for their assessment and treatment techniques. Three focus group interviews were conducted with physical therapists and physical therapist students, as well as a retrospective case note audit of 51 patients who had undergone surgery for hip fracture. Beyond early mobilization and a thorough day 1 postoperative assessment, great variability in what was considered to be best practice management was displayed. Senior physical therapists considered previous clinical experience to be more important than available research evidence, and junior physical therapists modeled their behavior on that of senior physical therapists. The amount of therapy provided to patients during their acute inpatient stay varied considerably, and none of the patients audited were seen on every day of their admission. Current physical therapist management in the acute setting for patients following hip fracture varies and is driven by system pressures as opposed to evidence-based practice.


Impact on hypertension reclassification by Ambulatory Blood Pressure Monitoring (ABPM) according to the V Brazilian Guidelines on ABPM.
New recommendations on reference values for normal test results in ambulatory blood pressure monitoring (ABPM) were proposed by the V Brazilian Guidelines on Ambulatory Blood Pressure Monitoring, based mainly on the IDACO study. This epidemiological study is aimed at evaluating the impact of adopting these new standards in an arterial hypertension referral center. The results of 1,567 ABPM tests carried out between 2005 and 2010 were analyzed; 481 patients were excluded from the sample for not meeting minimum quality criteria of the test. Reference values from the IV Brazilian Guidelines on ABPM (2005) were used for the classification of these tests regarding the abnormality and compared with the changes proposed by the V Brazilian Guidelines on ABPM (2011). Statistical analysis was performed by Pearson's chi-square method and p values < 0.05 were considered statistically significant. For the 1,086 tests evaluated, there was a significant difference in the proportion of patients with altered ABPM results, especially for the variable systolic pressure in the sleeping period: 49% when adopting the cutoff values of 2005 and 71% when adopting the values of 2011, with statistical significance, p < 0.0001. The recommendations of the new guidelines had a great impact on the hypertension classification by ABPM test results in the study population. The question of thresholds of these tests for therapeutic targets of patients known to be hypertensive is still open and requires further studies, preferably national ones, for better definition of the subject.


Thermodynamics-based rational design of DNA block copolymers for quantitative detection of single-nucleotide polymorphisms by affinity capillary electrophoresis.
Diblock copolymers composed of allele-specific oligodeoxyribonucleotide (ODN) and poly(ethylene glycol) (PEG) are used as an affinity probe of free-solution capillary electrophoresis to quantitatively detect single-base substitutions in genetic samples. During electrophoresis, the probe binds strongly to a wild-type single-stranded DNA analyte (WT) through hybridization, while it binds weakly to its single-base-mutated DNA analyte (MT) due to a mismatch. Complex formation with the probe augments the hydrodynamic friction of either analyte, thereby retarding its migration. The difference in affinity strength leads to separation of the WT, MT, and contaminants, including the PCR primers used for sample preparation. The optimal sequence of the probe's ODN segment is rationally determined in such a way that the binding constant between the ODN segment and MT at the capillary temperature is on the order of 10(6) M(-1). The validity of this guideline is verified using various chemically synthesized DNA analytes, as well as those derived from a bacterial genome. The peak area ratio of MT agrees well with its feed ratio, suggesting the prospective use of the present method in SNP allele frequency estimation.


An Updated Review on Marine Anticancer Compounds: The Use of Virtual Screening for the Discovery of Small-Molecule Cancer Drugs.
Marine secondary metabolites are a promising source of unexploited drugs that have a wide structural diversity and have shown a variety of biological activities. These compounds are produced in response to the harsh and competitive conditions that occur in the marine environment. Invertebrates are considered to be among the groups with the richest biodiversity. To date, a significant number of marine natural products (MNPs) have been established as antineoplastic drugs. This review gives an overview of MNPs, both in research or clinical stages, from diverse organisms that were reported as being active or potentially active in cancer treatment in the past seventeen years (from January 2000 until April 2017) and describes their putative mechanisms of action. The structural diversity of MNPs is also highlighted and compared with the small-molecule anticancer drugs in clinical use. In addition, this review examines the use of virtual screening for MNP-based drug discovery and reveals that classical approaches for the selection of drug candidates based on ADMET (absorption, distribution, metabolism, excretion, and toxicity) filtering may miss potential anticancer lead compounds. Finally, we introduce a novel and publically accessible chemical library of MNPs for virtual screening purposes.


[Phenotypic changes of mast cells during hepatocarcinogenesis in rats].
Sequential observation was made to detect the phenotypic change of mast cells during the process of dimethylaminoazobenzene-induced hepatocarcinogenesis in rats by cytochemistry, immunocytochemistry and electron microscopy. The number of mast cells in the liver tissues increased in various extent 4 weeks following the exposure to carcinogen. The increasing tendency was speeded up as time proceeded. At the 16th week, the number of mast cells reached to seven times as that in the normal rats. The mast cells appeared in the early stage of carcinogenesis were connective tissue mast cells (CTMC). From the 16th week on, mucosal mast cell (MMC) could be detected in some animals, but its number was much less than that of CTMC. In the late stages of experiment, the number of mast cells increased markedly, which showed positive reaction by ARMCP I and ARMCP II staining respectively and were to be brown or blue in color by combined Alcian blue. PAS. ABC staining. At the same time, an inter mediate type of mast cell between MMC and CTMC, light brown in color was also observed. Immunocytochemically, these cells presented ARMCP II positive reactivity. In various kinds of liver tumours formed at 26th week, mast cells occurred in a quite varying amount in different animals. No relationship was found between the amount of mast cells and the cell type or differentiation of the tumours, while the phenotypic distribution of mast cells was closely correlated with the histological type. The development and ultrastructural characterization of mast cells in the portal tracts and sinusoids were also investigated.


Deciphering CAPTCHAs: what a Turing test reveals about human cognition.
Turning Turing's logic on its head, we used widespread letter-based Turing Tests found on the internet (CAPTCHAs) to shed light on human cognition. We examined the basis of the human ability to solve CAPTCHAs, where machines fail. We asked whether this is due to our use of slow-acting inferential processes that would not be available to machines, or whether fast-acting automatic orthographic processing in humans has superior robustness to shape variations. A masked priming lexical decision experiment revealed efficient processing of CAPTCHA words in conditions that rule out the use of slow inferential processing. This shows that the human superiority in solving CAPTCHAs builds on a high degree of invariance to location and continuous transforms, which is achieved during the very early stages of visual word recognition in skilled readers.


A genomewide screen for autism susceptibility loci.
We report the analysis of 335 microsatellite markers genotyped in 110 multiplex families with autism. All families include at least two "affected" siblings, at least one of whom has autism; the remaining affected sibs carry diagnoses of either Asperger syndrome or pervasive developmental disorder. Affected sib-pair analysis yielded multipoint maximum LOD scores (MLS) that reach the accepted threshold for suggestive linkage on chromosomes 5, X, and 19. Nominal evidence for linkage (point-wise P<.05) was obtained on chromosomes 2, 3, 4, 8, 10, 11, 12, 15, 16, 18, and 20, and secondary loci were found on chromosomes 5 and 19. Analysis of families sharing alleles at the putative X chromosomal linked locus and one or more other putative linked loci produced an MLS of 3.56 for the DXS470-D19S174 marker combination. In an effort to increase power to detect linkage, scan statistics were used to evaluate the significance of peak LOD scores based on statistical evidence at adjacent marker loci. This analysis yielded impressive evidence for linkage to autism and autism-spectrum disorders with significant genomewide P values <.05 for markers on chromosomes 5 and 8 and with suggestive linkage evidence for a marker on chromosome 19.


Hsf4 counteracts Hsf1 transcription activities and increases lens epithelial cell survival in vitro.
The interplay between Hsf4 and Hsf1 plays an important role in the regulation of lens homeostasis. However, the mechanism of the intermolecular association involved is still unclear. In this paper, we find that reconstitution of Hsf4b into Hsf4-/- lens epithelial (mLEC/Hsf4-/-) cells can simultaneously downregulate Hsp70 expression and upregulate the expression of small heat shock proteins Hsp25 and αB-crystallin at both RNA and protein levels. ChIP assay results indicate Hsf4b, which binds to the promoters of Hsp90α, Hsp70.3, Hsp25 and αB-crystallin but not Hsp70.1, can inhibit Hsf1 binding to Hsp70.3 promoter and the heat shock mediated Hsp70 promoter activity by reducing Hsf1 protein expression. Hsf4b N-terminal hydrophobic region can interact with Hsf1 N-terminal hydrophobic region. Their interaction impairs Hsf1's intramolecular interaction between the N- and C-terminal hydrophobic regions, leading to Hsf1's cytosolic retention and protein degradation. Both lysosome inhibitors (chloroquine, pepstatin A plus E64d) and proteasome inhibitor MG132 can inhibit Hsf4-mediated Hsf1 protein degradation, but MG132 can induce Hsf1 activation as well. Upregulation of Hsf4b can significantly inhibit cisplatin and staurosporine induced lens epithelial cell apoptosis through direct upregulation of Hsp25 and αB-crystallin expression. Taken together, our results imply that upregulation of Hsf4b modulates the expression pattern of heat shock proteins in lens tissue by either directly binding to their promoters or promoting Hsf1 protein degradation. Moreover, upregulation of Hsf4b protects lens cell survival by upregulating anti-apoptotic pathways. These studies reveal a novel regulatory mechanism between Hsf1 and Hsf4b in modulating lens epithelial cell homeostasis.


The nurse as a social worker.
In the early twentieth century the role of the public health nurse (PHN) was expanding to meet the needs of individuals, families, and communities. Among the ideas discussed in the early nursing literature was that of the social nurse. This role was conceptualized as a combined nurse and social worker. Much of the PHNs work was with people of the poorer classes who needed assistance with both the medical aspects of their disease, as well as help with social needs such as food, rent money, and bedding. In this reprint from The Visiting Nurse Quarterly (1911), Ellen La Motte, Nurse-in-Chief with the Tuberculosis Division of the Baltimore Health Department, argued for this combined role based on economic efficiency. As she pointed out, it was "economic waste" to have two sets of workers going into homes when the nurse was capable, with additional education, of carrying out both roles. Additionally, from La Motte's perspective the only way the nurse could move beyond her role as the "physician's handmaiden" was through "social training" that prepared her for a broader scope of practice than that received in the hospital-based nursing schools. In the end, however, she left it open to nurses as "reasonable beings" to determine the direction the profession would take on this issue. La Motte's words provide historical context for issues contemporary public health nursing leaders are addressing, such as inter-professional boundaries between nurses and public health workers, reductions in the public health workforce, and economic constraints faced by the healthcare system. The Institute of Medicine mandate for inter-professional practice within the healthcare system, however, presents opportunities for exploring new roles and practice models for nurses in conjunction with our partners in public health.


[Nitrogen and protein content analysis of human milk, diurnality vs nocturnality].
Breast milk is changing with the progression of lactation and during a 24-h period. To determine the effect of diurnality or nocturnality on total nitrogen and protein content of the breast milk. We collected human milk samples from health mothers living throughout Community of Extremadura (Spain) from January 2008 to December 2008 with less than two months of lactation. We divided the samples in three groups: calostral group (1-5 days postpartum), transitional group (6-15 days postpartum) and mature group (> 15 days postpartum). All samples were stored in a freezer at -80 ºC. We considered as day period between 08:00-20:00 h and night period 20:00-08:00 h. Analysis of the human milk samples was based on the Kjeldahl method. Protein contents were calculated from total nitrogen x 6,25. The statistical analysis of the data was descriptive (mean ± standard deviation) and inferential (T-Student test). No differences (P > 0,05) were found to exist among the contents of individual human milk samples. The mean contents of each component were as follows: Total nitrogen of calostral, transitional and mature group was 0,30 ± 0,06 g/dL (night period), 0,29 ± 0,05 g/dL (day period); 0,26 ± 0,04 g/dL (night period), 0,25 ± 0,04 g/dL (day period); 0,22 ± 0,05 g/dL (night period), 0,20 ± 0,04 g/dL (day period) respectively, in this mature group with a statistical variation (P < 0,05). Protein content of calostral, transitional and mature group was 1,88 ± 0,4 g/dL (night period), 1,81 ± 0,3 g/dL (day period); 1,62 ± 0,3 g/dL (night period), 1,59 ± 0,3 g/dL (day period); 1,35 ± 0,3 g/dL (night period), 1,26 ± 0,3 g/dL (day period) respectively, in this mature group with a statistical variation (P < 0,05). Although we observed differences in the nitrogen and protein content during the individual stages of lactation, it is just in the population of mature lactating women, where the components analyzed varied significantly between day and night.


TwinCons: Conservation score for uncovering deep sequence similarity and divergence.
We have developed the program TwinCons, to detect noisy signals of deep ancestry of proteins or nucleic acids. As input, the program uses a composite alignment containing pre-defined groups, and mathematically determines a 'cost' of transforming one group to the other at each position of the alignment. The output distinguishes conserved, variable and signature positions. A signature is conserved within groups but differs between groups. The method automatically detects continuous characteristic stretches (segments) within alignments. TwinCons provides a convenient representation of conserved, variable and signature positions as a single score, enabling the structural mapping and visualization of these characteristics. Structure is more conserved than sequence. TwinCons highlights alternative sequences of conserved structures. Using TwinCons, we detected highly similar segments between proteins from the translation and transcription systems. TwinCons detects conserved residues within regions of high functional importance for the ribosomal RNA (rRNA) and demonstrates that signatures are not confined to specific regions but are distributed across the rRNA structure. The ability to evaluate both nucleic acid and protein alignments allows TwinCons to be used in combined sequence and structural analysis of signatures and conservation in rRNA and in ribosomal proteins (rProteins). TwinCons detects a strong sequence conservation signal between bacterial and archaeal rProteins related by circular permutation. This conserved sequence is structurally colocalized with conserved rRNA, indicated by TwinCons scores of rRNA alignments of bacterial and archaeal groups. This combined analysis revealed deep co-evolution of rRNA and rProtein buried within the deepest branching points in the tree of life.


Use of a collagen coated vicryl tube in reconstruction of the porcine esophagus.
This experimental study assessed the use of a collagen-coated vicryl mesh tube to reconstruct the esophagus in growing piglets. Initial experiments, in which a segment of thoracic esophagus was excised and replaced by this prosthetic tube, resulted in all the animals succumbing to mediastinitis within the first 3-4 days. This was shown, at post-mortem, to be due to leakage of the prosthesis, secondary to acid reflux, resulting in dissolution of the prosthesis. The collagen-coated vicryl mesh was thereafter treated with glutaraldehyde and in-vitro studies showed that the glutaraldehyde-treated material exhibited a higher resistance to 0.01 M HCl at pH 2.0, when compared with untreated material. In addition, the glutaraldehyde-treated material also showed an increased resistance to digestion by bacterial collagenase. Further experiments to reconstruct the cervical esophagus in growing piglets were performed using the revised glutaraldehyde cross-linked collagen coated vicryl mesh tube. All the animals survived the procedure and the prosthesis were leakproof immediately post-operatively. The animals, however, developed severe stenosis at a mean of 11 days post-operatively. Attempts at neo-epithelialization of the graft were seen histologically. There was considerable granulation tissue and scar tissue formation on the mediastinal aspect of the graft. It is suggested that collagen coated vicryl mesh tube may find applications in the treatment of esophageal atresia only if the problem of stenosis of the prosthesis can be solved.


Synthesis, antimicrobial and antineoplastic activities for agelasine and agelasimine analogs with a beta-cyclocitral derived substituent.
Agelasines and agelasimines are antimicrobial and cytotoxic purine derivatives isolated from marine sponges (Agelas sp.). We have synthesized structurally simplified analogs of these natural products starting from beta-cyclocitral. The novel compounds were found to be strong inhibitors of a wide variety of pathogenic microorganisms (incl. Mycobacterium tuberculosis) as well as cancer cell lines. The biological activities were generally in the same range as those previously found for the structurally more complex agelasines and agelasimines isolated in small amounts from natural sources. We also report for the first time that agelasine and agelasimine analogs inhibit growth of protozoa (Acanthamoeba castellanii and Acanthamoeba polyphaga). Acanthamoeba keratitis is an increasingly common and severe corneal infection, closely associated with contact lens wear.


Factors associated with readmission to adolescent psychiatric care.
The aim of this study is to identify factors associated with readmission to adolescent psychiatric inpatient care, in the context of a relapsing major mental illness. Data were obtained from 71 patients admitted to an adolescent psychiatric inpatient unit over a 2-year period. Patients who were rehospitalized within 12 months of discharge were compared with patients who had only one admission during the study period with respect to diagnosis, age of first admission, history of child sexual abuse (CSA) and the events precipitating admission. Medication non-adherence and a history of CSA were positively and independently associated with relapse readmission, while a trend emerged among readmitted patients toward younger age at first psychiatric hospitalization. A negative association was found between readmission and the experience of personal loss. Readmission was not related to DSM-IV axis I or axis II diagnosis, including substance abuse. The association, in an adolescent sample, of medication non-adherence and relapse readmission is consistent with findings from numerous adult studies, as is the trend toward younger age of first admission. The relationship of a history of CSA to readmission has attracted little previous research and the finding of a positive association suggests that further investigation of a history of CSA in this context is warranted. The implications of these findings and suggestions for further research are discussed.


Deficient aldehyde dehydrogenase 2 is associated with increased risk for esophageal squamous cell carcinoma in the presence of gastric hypochlorhydria.
In Orientals, deficient aldehyde dehydrogenase 2 (ALDH2) is associated with an increased risk for esophageal squamous cell carcinoma (ESCC). The local metabolism of carcinogenic acetaldehyde in the upper gastrointestinal tract could be involved in the association, but the underlying mechanism has not been fully elucidated. Since an anacidic stomach can promote bacteria-catalyzed local acetaldehyde production, the gastric acid level could also affect acetaldehyde metabolism. This study investigated whether ALDH2-related susceptibility to ESCC differs depending on the gastric secretion level. Sixty-two patients with ESCC and sex- and age-matched normal controls were enrolled in this study. ALDH2 polymorphism was analyzed by polymerase chain-restriction fragment length polymorphism, and those with an inactive allele (ALDH2-1/2-2 or ALDH2-2/2-2) were defined as ALDH2 deficient. Gastrin-stimulated acid output was assessed by endoscopic gastrin test and hypochlorhydria was defined as 0.6 mEq/10 min or lower. Multiple logistic regression analyses were used to adjust for other potential confounders. ALDH2 deficiency or hypochlorhydria was more prevalent in ESCC compared with controls and both showed increased independent associations with ESCC in multivariate analysis. Stratified analysis by the gastric acid secretion level revealed that the associations between the ALDH2 genotype and ESCC differed according to the individual gastric acid secretion levels and that ALDH2 deficiency was a significant risk factor for ESCC exclusively in individuals with hypochlorhydria with an odds ratio (95% confidence interval): 5.0 (1.2-21.2). Microbial production of carcinogen acetaldehyde in the presence of gastric hypochlorhydria is most probably involved in the mechanism of ALDH2-related susceptibility to ESCC.


Epidemiology of nocturnal enuresis in basic schoolchildren in Aden Governorate, Yemen.
Nocturnal enuresis is a common problem among children and adolescents. Data regarding this problem in schoolchildren in Yemen are scarce. This study was aimed to determine the prevalence of nocturnal incontinence in Aden school going children, describe its severity and identify the relation between nocturnal enuresis with personal and family characteristics. The study was a cross-sectional survey conducted on the public basic schoolchildren in Aden, Yemen, in the period November 2007-April 2009. A random, multistage sample of 890 students was taken from four districts in eight schools and divided into two strata: males and females. Data were obtained by using pre-recorded questionnaire, which was completed by parents. The response rate was 73.7% (656 students); 113 (17.2%) cases of nocturnal enuresis were encountered. Nocturnal enuresis decreased by age from 31.5% at 6-8 years to 8.7% at 15+ years (P < 0.05). Primary nocturnal enuresis affected 76.1%, of which the majority of children were bedwetting every night. Positive family history of nocturnal enuresis, deeper sleep, daytime enuresis, tea drinking, being non working father or with less education showed significant association with the occurrence of enuresis in the students. Stressful events in the previous 6 months of the study were twice more frequently noted. The study concluded that the prevalence of nocturnal enuresis in Aden public school children and its associated factors are almost comparable with that reported in epidemiological studies from various countries. Health education will encourage the parents to be aware, cope with this problem and seek appropriate medical advice.


Data Assimilation Methods for Neuronal State and Parameter Estimation.
This tutorial illustrates the use of data assimilation algorithms to estimate unobserved variables and unknown parameters of conductance-based neuronal models. Modern data assimilation (DA) techniques are widely used in climate science and weather prediction, but have only recently begun to be applied in neuroscience. The two main classes of DA techniques are sequential methods and variational methods. We provide computer code implementing basic versions of a method from each class, the Unscented Kalman Filter and 4D-Var, and demonstrate how to use these algorithms to infer several parameters of the Morris-Lecar model from a single voltage trace. Depending on parameters, the Morris-Lecar model exhibits qualitatively different types of neuronal excitability due to changes in the underlying bifurcation structure. We show that when presented with voltage traces from each of the various excitability regimes, the DA methods can identify parameter sets that produce the correct bifurcation structure even with initial parameter guesses that correspond to a different excitability regime. This demonstrates the ability of DA techniques to perform nonlinear state and parameter estimation and introduces the geometric structure of inferred models as a novel qualitative measure of estimation success. We conclude by discussing extensions of these DA algorithms that have appeared in the neuroscience literature.


Fibrinolytic Shutdown Is Associated With Intraoperative Thrombosis and Hemorrhage During Visceral Transplant.
Background and Objective. End-stage liver disease is characterized by a precarious imbalance of hemostasis. Detrimental consequences of hypofibrinolysis, also known as fibrinolytic shutdown, have been recently demonstrated, and its significance in visceral (ie, an allograft that contains the intestine) transplant remains unknown. Design and Setting. To fill this gap, following institutional review board approval, this retrospective study included 49 adult recipients of visceral allografts (14 "visceral allograft without the liver" and 35 "multivisceral" with the liver) transplanted between 2010 and 2018 in a single university hospital, and for whom pre-incisional thromboelastography was available. Based on percent clot lysis 30 minutes after maximal amplitude, patients were stratified into 3 fibrinolysis phenotypes: fibrinolytic shutdown, physiologic fibrinolysis, and hyperfibrinolysis. Results. Fibrinolytic shutdown occurred in 57% of patients, with higher incidence in recipients of multivisceral transplant (69%) compared with visceral allograft without liver (29%) allografts (P = .04). Fibrinolytic shutdown was associated with an increase in both intraoperative thrombosis and hemorrhage. Intraoperative thrombosis (18%) occurred only with multivisceral transplant, and accounted for 36% of in-hospital mortality. A clinically meaningful reduction in incidence of intraoperative thrombosis was noted in recipients who received intravenous heparin thromboprophylaxis. Logistic regression identified pretransplant platelet count as a risk factor for fibrinolytic shutdown (odds ratio = 0.992, 95% confidence interval = [0.984-0.998]; χ2 = 7.8, P = .005). Conclusions. This study highlights fibrinolytic shutdown as a dominant and clinically important feature of the hemostatic imbalance in recipients undergoing visceral transplantation.


A Brassica cDNA clone encoding a bifunctional hydroxymethylpyrimidine kinase/thiamin-phosphate pyrophosphorylase involved in thiamin biosynthesis.
We report the characterization of a Brassica napus cDNA clone (pBTHI) encoding a protein (BTHI) with two enzymatic activities in the thiamin biosynthetic pathway, thiamin-phosphate pyrophosphorylase (TMP-PPase) and 2-methyl-4-amino-5-hydroxymethylpyrimidine-monophosphate kinase (HMP-P kinase). The cDNA clone was isolated by a novel functional complementation strategy employing an Escherichia coli mutant deficient in the TMP-PPase activity. A biochemical assay showed the clone to confer recovery of TMP-PPase activity in the E. coli mutant strain. The cDNA clone is 1746 bp long and contains an open reading frame encoding a peptide of 524 amino acids. The C-terminal part of BTH1 showed 53% and 59% sequence similarity to the N-terminal TMP-PPase region of the bifunctional yeast proteins Saccharomyces THI6 and Schizosaccharomyces pombe THI4, respectively. The N-terminal part of BTH1 showed 58% sequence similarity to HMP-P kinase of Salmonella typhimurium. The cDNA clone functionally complemented the S. typhimurium and E. coli thiD mutants deficient in the HMP-P kinase activity. These results show that the clone encodes a bifunctional protein with TMP-PPase at the C-terminus and HMP-P kinase at the N-terminus. This is in contrast to the yeast bifunctional proteins that encode TMP-PPase at the N-terminus and 4-methyl-5-(2-hydroxyethyl)thiazole kinase at the C-terminus. Expression of the BTH1 gene is negatively regulated by thiamin, as in the cases for the thiamin biosynthetic genes of microorganisms. This is the first report of a plant thiamin biosynthetic gene on which a specific biochemical activity is assigned. The Brassica BTH1 gene may correspond to the Arabidopsis TH-1 gene.


Association of Race and Ethnicity With Glycemic Control and Hemoglobin A1c Levels in Youth With Type 1 Diabetes.
Health disparities in the clinical presentation and outcomes among youth with type 1 diabetes exist. Long-term glycemic control patterns in racially/ethnically diverse youth are not well described. To model common trajectories of hemoglobin A1c (HbA1c) among youth with type 1 diabetes and test how trajectory group membership varies by race/ethnicity. Longitudinal cohort study conducted in 5 US locations. The analysis included data from 1313 youths (aged <20 years) newly diagnosed in 2002 through 2005 with type 1 diabetes in the SEARCH for Diabetes in Youth study (mean [SD] age at diabetes onset, 8.9 [4.2] years) who had 3 or more HbA1c study measures during 6.1 to 13.3 years of follow-up. Data were analyzed in 2017. Self-reported race/ethnicity. Hemoglobin A1c trajectories identified through group-based trajectory modeling over a mean (SD) of 9.0 (1.4) years of diabetes duration. Multinomial models studied the association of race/ethnicity with HbA1c trajectory group membership, adjusting for demographic characteristics, clinical factors, and socioeconomic position. The final study sample of 1313 patients was 49.3% female (647 patients) with mean (SD) age 9.7 (4.3) years and mean (SD) disease duration of 9.2 (6.3) months at baseline. The racial/ethnic composition was 77.0% non-Hispanic white (1011 patients), 10.7% Hispanic (140 patients), 9.8% non-Hispanic black (128 patients), and 2.6% other race/ethnicity (34 patients). Three HbA1c trajectories were identified: group 1, low baseline and mild increases (50.7% [666 patients]); group 2, moderate baseline and moderate increases (41.7% [548 patients]); and group 3, moderate baseline and major increases (7.5% [99 patients]). Group 3 was composed of 47.5% nonwhite youths (47 patients). Non-Hispanic black youth had 7.98 higher unadjusted odds (95% CI, 4.42-14.38) than non-Hispanic white youth of being in the highest HbA1c trajectory group relative to the lowest HbA1c trajectory group; the association remained significant after full adjustment (adjusted odds ratio of non-Hispanic black race in group 3 vs group 1, 4.54; 95% CI, 2.08-9.89). Hispanic youth had 3.29 higher unadjusted odds (95% CI, 1.78-6.08) than non-Hispanic white youth of being in the highest HbA1c trajectory group relative to the lowest HbA1c trajectory group; the association remained significant after adjustment (adjusted odds ratio of Hispanic ethnicity in group 3 vs group 1, 2.24; 95% CI, 1.02-4.92). In stratified analyses, the adjusted odds of nonwhite membership in the highest HbA1c trajectory remained significant among male patients and youth diagnosed at age 9 years or younger, but not female patients and youth who were older than 9 years when they were diagnosed (P for interaction = .04 [sex] and .02 [age at diagnosis]). There are racial/ethnic differences in long-term glycemic control among youth with type 1 diabetes, particularly among nonwhite male patients and nonwhite youth diagnosed earlier in life.


Immunohistochemical evidence for preproenkephalin A synthesis in human retinoblastoma.
Human retinoblastoma contains clusters of cells immunoreactive for methionine-enkephalin and methionine-enkephalin-arginine-phenylalanine. Some tumour cells also exhibited methionine-enkephalin-arginine-glycine-leucine-like immunoreactivity. The results are in agreement with those obtained with similar testing of neuroblastoma cell cultures. It is concluded that some human retinoblastoma cells are capable of synthesizing preproenkephalin A or parts of this molecule.


Nitrogen and Sulfur Doped Carbon Dots from Amino Acids for Potential Biomedical Applications.
Nitrogen (N-) and sulfur (S-) doped carbon dots (CDs) were synthesized in a single step in a few min, 1-4 min via microwave technique from five different types of amino acids viz. Arginine (A), Lysine (L), Histidine (H), Cysteine (C), and Methionine (M). These amino acid derived N- and/or S- doped CDs were found to be in spherical shapes with 5-20 nm particle size range determined by Transition Electron Microscope (TEM) images and Dynamic Light Scattering (DLS) measurements. Thermal degradation, functional groups, and surface potential of the CDs were determined by Thermogravimetric Analysis (TGA), FT-IR spectroscopy, and zeta potential measurements, respectively. Although the zeta potential value of Cysteine derived CD (C-CD) was measured as -7.45±1.32 mV, the zeta potential values of A-CD, L-CD, H-CD, and M-CD particles were measured as +2.84±0.67, +2.61±1.0, +4.10±1.50 and+2.20±0.60 mV, respectively. Amongst the CDs, C- CDs was found to possess the highest quantum yield, 89%. Moreover, the blood compatibility test of CDs, determined with hemolysis and blood clotting tests was shown that CDs at 0.25 mg/mL concentration, CDs has less than 5% hemolysis ratio and higher than 50% blood clotting indexes. Furthermore, A-CD was modified with polyethyleneimine (PEI) and was found that the zeta potential values was increased to +34.41±4.17 mV (from +2.84±0.67 mV) inducing antimicrobial capability to these materials. Minimum Inhibition Concentration (MIC) of A-CD dots was found as 2.5 mg/mL whereas the PEI modified A-CDs, A-CD-PEI was found as 1 mg/mL against Escherichia coli ATCC 8739 (gram -) and Staphylococcus aureus ATCC 6538 (gram +) bacteria strains signifying the tunability of CDs.


Cloning and expression of a Salmonella enteritidis fimbrin gene in Escherichia coli.
A gene bank of DNA from a human isolate of Salmonella enteritidis was constructed in the cosmid pHC79 in Escherichia coli HB101. Five clones containing 35- to 45-kilobase inserts of S. enteritidis DNA reacted in colony immunoblot assays with a polyclonal antiserum prepared against purified S. enteritidis fimbriae. Electron microscopy showed that none of the five fimbrin-producing clones produced fimbriae, yet radioimmunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis located the 14,400-molecular-weight S. enteritidis in the outer membrane fraction of three of the clones and in the periplasmic fraction of all five clones. By using an oligonucleotide probe homologous to the 5' region of the fimbrin structural gene, the fimbrin gene was located on a 5.3-kilobase HindIII fragment. In vitro transcription-translation analysis verified that this HindIII fragment subcloned into plasmid pTZ18R produced unprocessed S. enteritidis fimbrin of molecular weight 16,400. Dot blot hybridization against a selection of strains of the family Enterobacteriaceae indicated a limited distribution of the S. enteritidis fimbrin gene.


How functional groups influence the ROS generation and cytotoxicity of graphene quantum dots.
Herein we selectively deactivate the ketonic carbonyl, carboxylic, or hydroxyl groups on GQDs and compare the ROS generation ability of different GQD derivatives. The results indicate that the ROS generation ability of GQDs is closely related to the ketonic carbonyl groups. Removal of the oxygen functional groups on GQDs can increase the photostability and lower the photoinduced cytotoxicity.


[Endometrial and ovarian morphology in internal uterine endometriosis].
The ovaries and endometrium were examined in 66 patients with internal uterine endometriosis with various degrees of the process dissemination. A high incidence of combinations of uterine endometriosis with hyperplastic processes in the ovaries was revealed (72.7%) and a relatively low incidence of uterine body mucosa involvement (31.7%). The degree of endometriosis dissemination in the myometrium and the incidence of ovarian hyperplasia were in direct proportion. The authors emphasize the usefulness of morphologic studies of the ovaries in patients with internal uterine endometriosis.


Simultaneous Aurora-A/STK15 overexpression and centrosome amplification induce chromosomal instability in tumour cells with a MIN phenotype.
Genetic instability is a hallmark of tumours and preneoplastic lesions. The predominant form of genome instability in human cancer is chromosome instability (CIN). CIN is characterized by chromosomal aberrations, gains or losses of whole chromosomes (aneuploidy), and it is often associated with centrosome amplification. Centrosomes control cell division by forming a bipolar mitotic spindle and play an essential role in the maintenance of chromosomal stability.However, whether centrosome amplification could directly cause aneuploidy is not fully established. Also, alterations in genes required for mitotic progression could be involved in CIN.A major candidate is represented by Aurora-A/STK15 that associates with centrosomes and is overexpressed in several types of human tumour. Centrosome amplification were induced by hydroxyurea treatment and visualized by immunofluorescence microscopy. Aurora-A/STK15 ectopic expression was achieved by retroviral infection and puromycin selection in HCT116 tumour cells. Effects of Aurora-A/STK15 depletion on centrosome status and ploidy were determined by Aurora-A/STK15 transcriptional silencing by RNA interference. Changes in the expression levels of some mitotic genes were determined by Real time RT-PCR. We investigated whether amplification of centrosomes and overexpression of Aurora-A/STK15 induce CIN using as a model system a colon carcinoma cell line (HCT116). We found that in HCT116 cells, chromosomally stable and near diploid cells harbouring a MIN phenotype, centrosome amplification induced by hydroxyurea treatment is neither maintained nor induces aneuploidy. On the contrary, ectopic overexpression of Aurora-A/STK15 induced supernumerary centrosomes and aneuploidy. Aurora-A/STK15 transcriptional silencing by RNA interference in cells ectopically overexpressing this kinase promptly decreased cell numbers with supernumerary centrosomes and aneuploidy. Our results show that centrosome amplification alone is not sufficient to induce chromosomal instability in colon cancer cells with a MIN phenotype. Alternatively, centrosome amplification has to be associated with alterations in genes regulating mitosis progression such as Aurora-A/STK15 to trigger CIN.


The role of short-term oxygen administration in the prevention of intimal hyperplasia.
Intimal hyperplasia (IH) is the cause of most failed arteriovenous fistulas (AVFs), resulting in repeat procedures and leading to increased utilization of scarce health care resources. Our laboratory has previously demonstrated the role of supplemental oxygen in preventing IH and smooth muscle cell proliferation (SMCp) at an artery-to-graft anastomosis and at the deployment site of an intra-arterial stent. This study examines the effect of supplemental oxygen in preventing IH and SMCp in an AVF in a rabbit model. Ninety-six rabbits were randomized into four groups: group 1, control; group 2, no surgery with supplemental oxygen; group 3, AVF without supplemental oxygen; and group 4, AVF with supplemental oxygen. Rabbits receiving supplemental oxygen received 30% oxygen for up to 42 days. Specimens were collected in all groups at days 1, 3, 7, 21, 42, and 90. IH and SMCp were measured at the AVF site as well as in the artery and vein proximal and distal to the AVF. IH was first noted at day 7 and significantly increased through day 90 at all locations in the nonoxygen-supplemented groups. No significant IH was noted in the oxygen-supplemented group at any location or any time point. SMCp was noted at day 3 through day 21 in the nonoxygen-supplemented group, whereas almost no SMCp was noted in the oxygen-supplemented group at any location or time point. Without oxygen supplementation, SMCp begins at day 3 and is no longer noted at day 21 after creation of an AVF, whereas IH begins by day 7 and increases at least through day 90 after creation of an AVF. Forty-two days of 30% supplemental oxygen inhibits IH and SCMp after creation of an AVF. These data suggest a role for the short-term administration of low-dose O2 to prevent both IH and SMCp after creation of an AVF that may prolong patency and function.


Using Xenopus skin to study cilia development and function.
Cilia are prevalent biological structures that are important for cell signaling and for generating fluid flow (or motility). Cilia are found throughout biology from single-celled organisms to vertebrates, and many model systems have been employed for their analysis. Here, we describe the use of Xenopus larval skin as a system for the study of ciliogenesis and ciliary function. In particular, we describe basic molecular and embryological manipulations and imaging techniques that have proven particularly useful for understanding the polarized beating of cilia and the generation of directed fluid flow (Werner & Mitchell, 2012). However, these same tools have the potential to benefit a large number of cilia-related biological questions.


Enteroendocrine cells express functional Toll-like receptors.
Intestinal epithelial cells (IECs) provide a physical and immunological barrier against enteric microbial flora. Toll-like receptors (TLRs), through interactions with conserved microbial patterns, activate inflammatory gene expression in cells of the innate immune system. Previous studies of the expression and function of TLRs in IECs have reported varying results. Therefore, TLR expression was characterized in human and murine intestinal sections, and TLR function was tested in an IEC line. TLR1, TLR2, and TLR4 are coexpressed on a subpopulation of human and murine IECs that reside predominantly in the intestinal crypt and belong to the enteroendocrine lineage. An enteroendocrine cell (EEC) line demonstrated a similar expression pattern of TLRs as primary cells. The murine EEC line STC-1 was activated with specific TLR ligands: LPS or synthetic bacterial lipoprotein. In STC-1 cells stimulated with bacterial ligands, NF-kappaB and MAPK activation was demonstrated. Furthermore, the expression of TNF and macrophage inhibitory protein-2 were induced. Additionally, bacterial ligands induced the expression of the anti-inflammatory gene transforming growth factor-beta. LPS triggered a calcium flux in STC-1 cells, resulting in a rapid increase in CCK secretion. Finally, conditioned media from STC-1 cells inhibited the production of nitric oxide and IL-12 p40 by activated macrophages. In conclusion, human and murine IECs that express TLRs belong to the enteroendocrine lineage. Using a murine EEC model, a broad range of functional effects of TLR activation was demonstrated. This study suggests a potential role for EECs in innate immune responses.


Transgenic expression of the soybean apyrase in Lotus japonicus enhances nodulation.
The soybean apyrase, GS52, was previously characterized as an early nodulin that is expressed in roots and localized to the plasma membrane. Transgenic Lotus japonicus plants were constructed constitutively expressing the GS52 apyrase. Segregation and Southern-blot analysis identified four single-copy sense lines, several double-copy sense lines, and one double-copy antisense line for further analysis. The single- and double-copy sense gs52 L. japonicus lines had enhanced nodulation that correlated with expression of the transgene. The sense transgenic lines were also found to have increased infection thread formation and enhanced infection zone length when infected by Mesorhizobium loti, the natural symbiont of L. japonicus. The data presented show that expression of the GS52 apyrase can enhance nodulation in L. japonicus and points to an important role for this group of enzymes in nodulation.


Humility and its practice in nursing.
Following a personal experience of transformation as a result of washing the feet of a terminally ill patient, an exploratory study was undertaken to investigate nurses' experience of washing patients' feet. Seven postregistration student nurses participated in the study by washing the feet of as many patients as they could over a defined period of time. They were then interviewed about the experience. The transcribed interviews were analysed using the heuristic enquiry approach. Symbolically, washing feet is an act of humility. In washing feet in the manner required for this study I suggest that the nurses were practising beyond role definition of duty of care. As a result of this they experienced interconnectedness and changes in their relationship with the patients whose feet they had washed that could be interpreted as a response to humility.


Flapless surgery and its effect on dental implant outcomes.
The aim of this article was to review the current literature with regard to the efficacy and effectiveness of flapless surgery for endosseous dental implants. The available data were evaluated for short- and long-term outcomes. A MEDLINE search was conducted on studies published between 1966 and 2008. For the purpose of this review, only clinical (human) studies with five or more subjects were included, and clinical opinion papers were excluded. Clinical studies or reports were further rated in terms of the level or weight of evidence using criteria defined by the Oxford Center for Evidence-Based Medicine in 2001. The available data on flapless technique indicate high implant survival overall. The prospective cohort studies demonstrated approximately 98.6% (95% CI: 97.6 to 99.6) survival, suggesting clinical efficacy, while the retrospective studies or case series demonstrated 95.9% (95% CI: 94.8 to 97.0) survival, suggesting effective treatment. Six studies reported mean radiographic alveolar bone loss ranging from 0.7 to 2.6 mm after 1 year of implant placement. Intraoperative complications were reported in four studies, and these included perforation of the buccal or lingual bony plate. Overall, the incidence of intraoperative complications was 3.8% of reported surgical procedures. Flapless surgery appears to be a plausible treatment modality for implant placement, demonstrating both efficacy and clinical effectiveness. However, these data are derived from short-term studies with a mean interval of 19 months, and a successful outcome with this technique is dependent on advanced imaging, clinical training, and surgical judgment.


Direct effect of human immunodeficiency virus protease inhibitors on neutrophil function and apoptosis via calpain inhibition.
Impairment of neutrophil functions and high levels of apoptotic neutrophils have been reported in human immunodeficiency virus (HIV) patients. The aim of the present study was to investigate the direct in vitro effects of the different HIV protease inhibitors (PIs) on neutrophil functions and apoptosis and to explore their mechanisms of action. The effects of nelfinavir (NFV), saquinavir (SQV), lopinavir (LPV), ritonavir (RTV), and amprenavir (APV) in the range of 5 to 100 microg/ml on neutrophil function, apoptosis, and mu-calpain activity were studied. The neutrophil functions studied included superoxide production stimulated by 5 ng/ml phorbol myristate acetate, 5 x 10(-7) M N-formyl-methionyl-leucyl-phenylalanine, and 1 mg/ml opsonized zymosan; specific chemotaxis; random migration; and phagocytosis. Apoptosis was determined by DNA fragmentation, fluorescein isothiocyanate-annexin V binding, and nuclear morphology. All three neutrophil functions, as well as apoptosis, were similarly affected by the PIs. SQV and NFV caused marked inhibition and LPV and RTV caused moderate inhibition, while APV had a minor effect. mu-Calpain activity was not affected by the PIs in neutrophil lysate but was inhibited after its translocation to the membranes after cell stimulation. SQV, which was the most potent inhibitor of neutrophil functions and apoptosis, caused significant inhibition of calpain activity, while APV had no effect. The similar patterns of inhibition of neutrophil functions and apoptosis by the PIs, which coincided with inhibition of calpain activity, suggest the involvement of calpain activity in the regulation of these processes.


Thermo-optical effects in high-power ytterbium-doped fiber amplifiers.
We investigate the effect of temperature gradients in high-power Yb-doped fiber amplifiers by a numerical beam propagation model, which takes thermal effects into account in a self-consistent way. The thermally induced change in the refractive index of the fiber leads to a thermal lensing effect, which decreases the effective mode area. Furthermore, it is demonstrated that the thermal lensing effect may lead to effective multi-mode behavior, even in single-mode designs, which could possibly lead to degradation of the output beam quality.


Platelet-derived growth factor in glioblastoma-driver or biomarker?
The platelet-derived growth factor (PDGF) family of mitogens exerts vital functions during embryonal development, e.g. in the central nervous system, where PDGF drives the proliferation of oligodendrocyte precursors. PDGF and PDGF receptors are co-expressed in human glioblastoma (GBM). Whether an aberrant activation of the PDGF receptor pathway is a driving force in glioma development has remained an open question. In experimental animals, overexpression of PDGF has convincingly been shown to induce tumors, both in wild-type animals (marmoset, rat, mouse) and in mice with targeted deletions of suppressor genes, e.g. Tp53 or Ink4A. Targeting the PDGF receptor in tumor-bearing mice leads to growth inhibition and reversion of the transformed phenotype. Findings of PDGF receptor amplification or mutations in human GBM are strong indicators of a causative role of the PDGF receptor pathway. However, clinical trials using PDGF receptor antagonists have been disappointing. In conclusion, a PDGF receptor profile may be a biomarker for a subgroup of GBM originating from a PDGF receptor-responsive cell. Although compelling experimental and clinical evidence supports the notion that the PDGF receptor pathway is a driver in GBM, formal proof is still missing.


Prevalence and antimicrobial resistance of Salmonella isolated from two pork processing plants in Alberta, Canada.
This study investigated the frequency of Salmonella serovars on pig carcasses at various processing steps in two commercial pork processing plants in Alberta, Canada and characterized phenotypic and genotypic antimicrobial resistance (AMR) and PFGE patterns of the Salmonella isolates. Over a one year period, 1000 swab samples were collected from randomly selected pigs at two slaughter plants. Sampling points were: carcass swabs after bleeding (CSAB), carcass swabs after de-hairing (CSAD, plant A) or skinning (CSASk, plant B), carcass swabs after evisceration (CSAE), carcass swabs after pasteurization (CSAP, plant A) or washing (CSAW, plants B) and retail pork (RP). For plant A, 87% of CSAB and 8% of CSAE were positive for Salmonella while at plant B, Salmonella was recovered from 94% of CSAB and 10% of CSAE. Salmonella was not recovered from the RP samples at either plant, indicating that the plants used effective control measures. Salmonella enterica serovar Derby was the most common serotype (23%, 29/127) recovered in plant A and plant B (61%, 76/124). For plant A, 35% (45/127) of isolates were resistant to at least one antimicrobial. Five isolates (3.9%), 4 serovar Ohio strains and one serovar I:Rough-O:I,v:-, strain were simultaneously resistant to antimicrobials of very high (Category I), high (Category II), and medium (Category III) importance to human medicine. The 4 S. Ohio isolates were recovered from 3 different steps of pork processing on the same sampling day and displayed resistance to 5-7 antimicrobials, with all of them displaying resistance to ceftiofur and ceftriaxone (Category I). An I:Rough-O:l,v:- isolate, recovered on a different sampling day, was resistant to 7 antimicrobials that included resistance to ampicillin/clavulanic acid, ceftiofur and ceftriaxone (Category I). Salmonella strains isolated from plant A harbored 12 different AMR genes. The most prevalent genes were sul1, sul2, tet(A), tet(B), aadA, strA/strB, aac(3)IV and aphA1. For Salmonella isolates from plant B, 7 resistance genes were identified alone or in combination where tet(B) was found in 77 (62.3%) of the isolates. For plant A, 19 different PFGE subtypes of Salmonella isolates that displayed phenotypic and/or genotypic resistance were observed while 13 different PFGE subtypes were observed for plant B. The lack of detection of Salmonella on the surfaces of RP suggests that current pork processing practices can dramatically reduce Salmonella. Salmonella isolates from pig carcasses at various steps displayed multidrug resistance, including to those of very high importance in human medicine, which represent a public health concern.


Global analysis of the eukaryotic pathways and networks regulated by Salmonella typhimurium in mouse intestinal infection in vivo.
Acute enteritis caused by Salmonella is a public health concern. Salmonella infection is also known to increase the risk of inflammatory bowel diseases and cancer. Therefore, it is important to understand how Salmonella works in targeting eukaryotic pathways in intestinal infection. However, the global physiological function of Salmonella typhimurium in intestinal mucosa in vivo is unclear. In this study, a whole genome approach combined with bioinformatics assays was used to investigate the in vivo genetic responses of the mouse colon to Salmonella. We focused on the intestinal responses in the early stage (8 hours) and late stage (4 days) after Salmonella infection. Of the 28,000 genes represented on the array, our analysis of mRNA expression in mouse colon mucosa showed that a total of 856 genes were expressed differentially at 8 hours post-infection. At 4 days post-infection, a total of 7558 genes were expressed differentially. 23 differentially expressed genes from the microarray data was further examined by real-time PCR. Ingenuity Pathways Analysis identified that the most significant pathway associated with the differentially expressed genes in 8 hours post-infection is oxidative phosphorylation, which targets the mitochondria. At the late stage of infection, a series of pathways associated with immune and inflammatory response, proliferation, and apoptosis were identified, whereas the oxidative phosphorylation was shut off. Histology analysis confirmed the biological role of Salmonella, which induced a physiological state of inflammation and proliferation in the colon mucosa through the regulation of multiple signaling pathways. Most of the metabolism-related pathways were targeted by down-regulated genes, and a general repression process of metabolic pathways was observed. Network analysis supported IFN-γ and TNF-α function as mediators of the immune/inflammatory response for host defense against pathogen. Our study provides novel genome-wide transcriptional profiling data on the mouse colon mucosa's response to the Salmonella typhimurium infection. Building the pathways and networks of interactions between these genes help us to understand the complex interplay in the mice colon during Salmonella infection, and further provide new insights into the molecular cascade, which is mobilized to combat Salmonella-associated colon infection in vivo.


Testicular damage in children and adolescents treated for malignancy: a short review.
Significant advances have been made over recent decades in the treatment of childhood malignancies. These advances had an incredible cost, as an increasing number of young survivors suffer subfertility or infertility, because of the high sensitivity of testicular cells, especially the rapidly dividing germ cells, to cytotoxic drugs and irradiation. Therefore, the impact of treatment on future fertility is of significant concern, both to parents and patients. Assessment of fertility damage in childhood remains problematic. For post-pubertal males, semen analysis represents a good indicator of spermatogenesis and testicular function, and allows for sperm cryopreservation. The available method for prepubertal children is only gonadal tissue cryopreservation. This method is still experimental and raises ethical concerns. Ideally, a multidisciplinary team approach needs to be used in addressing the needs of fertility preservation for this population. Precise knowledge of these issues would help pediatric oncologists and endocrinologists to counsel their patients and inform them for factors and resources that may protect or preserve parenthood options in the future.


Fatty acid oxidation and the beta-oxidation complex in Mycobacterium leprae and two axenically cultivable mycobacteria that are pathogens.
Intact, non-growing Mycobacterium leprae, M. avium and M. microti oxidized a wide range of 1-14C-labelled fatty acids (C8 to C24) to 14CO2. Laurate (C12) was oxidized most rapidly, and its oxidation by M. leprae was inhibited by the antileprosy agents Dapsone, clofazamine and rifampicin. Key enzymes of beta-oxidation were detected in extracts from all three mycobacteria. All these activities (both in intact mycobacteria and the enzymes) were stimulated in M. avium grown in Dubos medium plus palmitate but activities in M. microti or M. avium grown either in Dubos medium with added liposomes or triolein, or in vivo were similar to those detected in the same strain grown in Dubos medium alone. M. avium could be grown in medium in which 95% of its fatty acyl elongase activity is acetyl-CoA dependent. In this medium growing M. avium organisms oxidized [1-14C]palmitate to 14CO2 but simultaneously elongated palmitate to C24 acids and even longer. Acetyl-CoA-dependent elongase activity is similar but clearly not identical to reversed beta-oxidation, but the exact point(s) of difference have not yet been identified.


Risk perception and sex behaviour in pregnancy and breastfeeding in high HIV prevalence settings: Programmatic implications for PrEP delivery.
HIV acquisition during pregnancy and breastfeeding significantly contributes toward paediatric HIV infection; however, little is known about risk behaviours in HIV-uninfected pregnant and postpartum women. We conducted twenty-six in-depth-interviews between July and December 2016 using a semi-structured interview guide among HIV-uninfected pregnant and recently postpartum women at-risk of HIV acquisition (defined as reporting ≥1 of the following: partner's serostatus unknown or HIV-infected, recent condomless sex in pregnancy, and/or alcohol use during pregnancy) who attended primary healthcare services. Our study contextualizes factors related to risky sexual behaviours during pregnancy and postpartum periods and assesses knowledge and hypothetical acceptability of pre-exposure prophylaxis (PrEP) in pregnancy. Translated and transcribed data were coded and analysed by three researchers using a thematic analysis approach. In interviews with HIV-uninfected pregnant/postpartum women at-risk of HIV acquisition, we identified common themes associated with sexual risk behaviours during pregnancy, including: lack of control over decisions in sex and condom use in pregnancy, low perceived risk (e.g. beliefs that their partner has the same HIV-negative serostatus), and socio-cultural beliefs around condom use during pregnancy (e.g. contact with sperm is essential for baby's development). PrEP knowledge was low among HIV-uninfected pregnant and breastfeeding women, and potential acceptability was good, though primary concerns were around the potential impact on the infant. While mothers presented a clear desire to protect themselves from HIV acquisition once pregnant, they also reported lack of control, and socio-cultural beliefs, like sex is good for the baby, that increased their risk of seroconversion. Mothers had limited PrEP awareness but reported hypothetical willingness to use PrEP because of concerns over HIV acquisition and onward mother to child transmission.


Testis imaging with 111In-labeled anticarcinoembryonic antigen monoclonal antibody: identification of carcinoembryonic antigen in normal germ cells.
A monoclonal antibody to carcinoembryonic antigen (CEA) labeled with 111In (Indacea) was used to image tumors in patients with colorectal cancer. The anti-CEA antibody has a high affinity (2.6 X 10(10) M-1) for CEA and does not cross-react with normal cross-reacting antigen, biliary glycoprotein-1, or tumor-extracted, CEA-related antigen. During the course of these studies, it was noted that a significant number of male patients (20 of 27, 74%) showed uptake of Indacea in the testes. In order to determine if the Indacea uptake was specific, 20 testicular specimens were analyzed by immunohistological methods using five different anti-CEA CEA monoclonal antibodies recognizing five different epitopes on CEA. In 18 cases (90%) germ cells were uniformly stained by all five antibodies. Fresh frozen testis tissue was homogenized in water and precipitated with 1.0 M perchloric acid. The supernatant contained a CEA-like material as measured by an enzyme immunoassay specific for CEA. The same supernatant was radiolabeled with 125I and immunoprecipitated with anti-CEA monoclonal antibodies. Sodium dodecyl sulfate:polyacrylamide gel electrophoresis of the immunoprecipitates revealed a single species (Mr 180,000) which was indistinguishable from CEA. This study documents the first description of CEA in the germ cells of normal testis. The CEA in the testis was accessible to circulating monoclonal antibodies in the majority of male patients tested.


Structural and biological studies on synthetic peptide analogues of a low-affinity calcium-binding site of skeletal troponin C.
We have synthesized four oligopeptides that are structural analogues of a low-affinity Ca2+-specific binding site (site II) of rabbit skeletal troponin C. One analogue (peptide 3) was a dodecapeptide with a sequence corresponding to the 12-residue Ca2+-binding loop (residues 63-74 in troponin C), two (peptides 4 and 5) were 23-residue in length, corresponding to residues 52-74 of the protein, and the fourth (peptide 6) was a 25-residue peptide corresponding to residues 50-74. All four peptides had one amino acid substitution within the 12-residue binding loop in which phenylalanine at position 10 was replaced by tyrosine to provide a marker for spectroscopic studies. In addition, peptides 3 and 4 each had a second substitution within the binding loop where glycine at position 6 was replaced by alanine. The second substitution was motivated by the conservation of glycine at the position in the Ca2+-binding loops of all four Ca2+-binding sites in troponin C. The peptides were characterized by their intrinsic fluorescence, ability to enhance the emission of bound Tb3+, affinity for Ca2+ and Tb3+, and circular dichroism. The affinity for Ca2+ was in the range 10-10(2) M-1, and the affinity for Tb3+ was in the range 10(4)-10(5) M-1. The binding constants of the longer peptides were several-fold larger than that of the dodecapeptide. With peptides 4 and 5, substitution of glycine by alanine at position 6 within the 12-residue loop decreased the affinity for Ca2+ by a factor of four, but had little effect on the affinity for Tb3+. However, the mean residue ellipticity of peptide 4 was substantially higher than that of peptide 5. Since peptide 4 differs from peptide 5 only in the substitution of glycine at position 6 in the loop segment, the conservation of glycine at that position may serve a role in providing a suitable secondary structure of the binding sites for interaction with troponin I. Peptides 4 and 6, when present in a large excess, mimic troponin C in regulating fully reconstituted actomyosin ATPase by showing partial calcium sensitivity and activation of the ATPase. Since these peptides are the smallest peptides containing the Ca2+-binding loop of site II, their biological activity suggests that a Ca2+-dependent binding site of troponin C for troponin I could be as short as the segment comprising residues 52-62.


A voluntary transfusion recipient registry in Korea as a database for blood group antibodies.
Several types of transfusion-related registries have been developed to improve patient outcomes and blood banks. In Korea, a transfusion program functioning as a blood group antibody database and a reference laboratory has been in operation since July 2013. This study was conducted to determine the current status of blood group antigens and antibodies in Korea and propose a model for registries in the field of transfusion medicine. Cases with unexpected red cell antibodies were registered online in the voluntary transfusion registry. Specific antigen-negative frequencies were calculated based on the recorded data. To determine the frequencies of RhCE antigens, data added via the Blood Information Sharing System were also analyzed. Data added to the registries between July 2013 and June 2022 were included in the analysis. Among 9,048 antibody cases registered from 29 hospitals, anti-E alone was identified most commonly, followed by anti-E and c, anti-C and e, anti-Lea, and anti-M (2,202, 1,792, 757, 618, and 383 cases, respectively). The frequencies of E-, E-c-, C-e-, Le(a-), and M- were 49.1%, 41.6%, 9.1%, 69.4%, and 21.8%, respectively. The distributions of antibodies and antigen frequencies were estimated through the transfusion registry. Antigen frequencies were calculated based on the results of antigen typing of red blood cell components performed at the time of issuing. The online transfusion registry serving as a blood group antibody database is useful for determining the frequencies of blood group antigens and antibodies.


Of Our Territory, We Know: social mapping experience for emergencies and disasters.
The article presents the project "Of Our Territory, We Know" (De Nosso Território Sabemos Nós), carried out in two communities in the mountain cities of the state of Rio de Janeiro that suffered the 2011 disaster, aiming at creating a device for online communication, associated with social cartography, seeking to establish a dialogue between the city management and citizens in a permanent and dynamic way. The device allows local needs to be visible for the creation of public policies. The implementation of the Extended Research Communities (Comunidades Ampliadas de Pesquisa), consisting of residents and their associations, participants of the municipal management, and researchers in a regime of ecology of knowledges, took place through cartographic workshops. The identification of the vulnerabilities and potentials of the communities, the emergence of memories and local knowledges allow the strengthening of community resilience. Therefore, the appraisal of the experience in the creation of social cartographies brings out the group's self-awareness. The mapping process demonstrated how unknown the territories are to the municipal management and even to their inhabitants.


Improving credibility of instructions in the balanced placebo design: a misattribution manipulation.
Research using the balanced placebo design seeks to differentiate the physiological and psychological effects of drinking alcohol. Questions regarding the validity of the design center about experimenter instructions, particularly in the antiplacebo cell at higher blood alcohol content (BAC) levels. This study tested the plausibility of two misattribution strategies designed to reduce the conflict between experimenter instructions and internal cues of drunkenness. Forty-two participants (BAC = .055) were told that they received no alcohol, with internal cues of drunkenness said to be produced by a (sham) second drug, a (placebo) tachistoscopic display, or no misattribution given. The placebo drug group reported less alcohol intoxication without reporting less physical impairment than the control or tachistoscopic groups. Doubt of instructions was expressed more frequently in the control group than in the placebo drug group. Mean time to first reported doubt of experimenter instructions was longer for the placebo drug group. A manipulation check designed to account for demand effects indicated that instituting the pharmacologic misattribution increased the success of the manipulation over the control group. Providing a credible attribution for internal symptoms of drunkenness makes experimenter's instructions more credible, improving the validity of the antiplacebo cell of the balanced placebo design.


Cloning of rat 17 beta-hydroxysteroid dehydrogenase type 2 and characterization of tissue distribution and catalytic activity of rat type 1 and type 2 enzymes.
17 beta-Hydroxysteroid dehydrogenases (17HSDs) are enzymes catalyzing the conversion between 17 beta-hydroxy- and 17-ketosteroids. Both estrogens and androgens possess their highest activity in the 17 beta-hydroxy form, and the enzymes, therefore, regulate the biological activity of sex hormones. In this study, we have characterized the complementary DNA (cDNA) for rat 17HSD type 2. The cDNA encodes a protein with a predicted mol wt of 42,010 Da. The protein has 77% similarity and 62% identity with the human 17HSD type 2 enzyme. Furthermore, the hydropathicity profiles of the enzymes are very similar. The two isozymes contain a putative transmembrane region close to the N-terminus. However, the rat isozyme lacks the two lysine-rich amino acid cluster present at the N- and C-terminals of human 17HSD type 2. The tissue distribution of the rat 17HSD type 1 and type 2 enzymes is very similar to that of the human enzymes. The highest expression of 17HSD type 2 was detected in the placenta. In addition, a 1.5-kilobase messenger RNA for the enzyme was detected in the small intestine, liver, and kidney of both sexes. The two messenger RNAs for rat 17HSD type 1 (1.4 and 1.7 kilobases) were highly expressed only in the ovary, and at very low concentrations in the kidney of both sexes. Transiently expressed rat 17HSD type 2 showed oxidative activity almost exclusively in cultured human embryonic kidney 293 cells, converting estradiol into estrone and testosterone into androstenedione, whereas the opposite was observed for the rat type 1 enzyme. The data suggest that similarly to the corresponding human isoforms, rat 17HSD type 2 is mostly involved in the oxidation of 17 beta-hydroxysteroids into their relatively inactive keto derivative in peripheral tissues, whereas rat 17HSD type 1 is mainly involved in the glandular biosynthesis of estradiol.