Anti-obesity effects of Laminaria japonica fermentation on 3T3-L1 adipocytes are mediated by the inhibition of C/EBP-α/β and PPAR-γ.
Obesity is global problem that contributes to disease, and is partly caused by fast-food, high-fat diets. Much attention has been focused on developing anti-obesity foods and chemical materials from natural sources. Seaweed has bioactive properties that influence immune activity and have anti-cancer and anti-obesity effects. Laminaria japonica is a widely consumed seaweed, and has been promoted as a health food in Korea. The bioactive properties of L. japonica include anti-cancer, anti-diabetic, and anti-inflammation effects. Most Laminaria japonica are distributed in a simple processing form such as drying, and their availability is very low. Therefore, various types of functional products can be developed if they can be applied to foods through functionalization using fermentation techniques. It is a structural problem that is the most problematic in seaweed processing. In this study, we used fermented Laminaria japonica. To increase physiological activity, fermentation treatment was performed to loosen the structure, thereby increasing the activity of the glycoprotein. First, we screened the anti-obesity potential of an L. japonica fermentation extract (LJF) using 3T3-L1 adipocyte cells. We determined cytotoxicity using an MTS assay and measured LJF for its ability to affect adipogenesis through glucose uptake, triglyceride levels, and Oil Red O staining. We confirmed that LJF inhibited adipocyte differentiation. CCAAT/enhancer-binding proteins α/β (C/EBP-α/β) and peroxisome proliferator-activated receptor-γ (PPAR-γ) are involved in the early and late stages of adipocyte differentiation. LJF significantly reduced the expression levels of C/EBP-α/β and PPAR-γ and decreased the concentration of adiponectin. Thus, our results suggest that LJF inhibits adipogenesis in 3T3-L1 cells, and may be valuable for its anti-obesity effects.


Old and new functions of proneural factors revealed by the genome-wide characterization of their transcriptional targets.
In the developing vertebrate nervous system, bHLH proneural factors such as Ascl1 are known to play important regulatory roles at different stages of the neurogenic differentiation process. In spite of the wealth of information gathered on the cellular functions of proneural factors, little was known of the molecular basis for their activities, and in particular of the identity of their target genes. The development of genomic approaches is making possible the characterization of transcriptional programs at an unprecedented scale. Recently, we have used a combination of genomic location analysis by ChIP-on-chip and expression profiling in order to characterize the proneural transcription program regulated by Ascl1 in the ventral telencephalon of the mouse embryonic brain. Our results demonstrate that Ascl1 directly controls successive steps of neurogenesis and provide a molecular frame for previously described Ascl1 functions. In addition, we uncovered an important but previously unrecognized role for Ascl1 in promoting the proliferation of neural progenitors. Here we discuss our recent findings and review them in light of efforts from other laboratories to characterize the transcriptional programs downstream various proneural factors.


Prospective study of oral teicoplanin versus oral vancomycin for therapy of pseudomembranous colitis and Clostridium difficile-associated diarrhea.
A prospective, randomized study comparing oral teicoplanin with oral vancomycin in the treatment of pseudomembranous colitis (PMC) and Clostridium difficile-associated diarrhea (CDAD) was performed. Teicoplanin was administered at a dosage of 100 mg twice a day for 10 days, and vancomycin was administered at a dosage of 500 mg four times a day for 10 days. CDAD was diagnosed by demonstrating both C. difficile and cytotoxin in the feces of symptomatic patients (more than three loose stools per day). The diagnosis of PMC was also based on colonoscopy. Cytotoxin assay and cultures were checked in all patients 7 to 10 days after discontinuation of therapy and 25 to 30 days thereafter. Of the 51 patients enrolled, 46 were judged to be assessable. Among these, 26 received teicoplanin and 20 received vancomycin. At enrollment, both groups were comparable in terms of age, sex, occurrence of PMC or CDAD, and previous antibiotic treatment. Eighteen of the 20 patients in the vancomycin group and 10 of the 26 patients in the teicoplanin group had previously undergone surgery (P = 0.0004). Treatment resulted in the clinical cure of 20 (100%) vancomycin and 25 (96.2%) teicoplanin patients (P = 0.56). After discontinuation of therapy, clinical symptoms recurred in four (20%) vancomycin patients and two (7.7%) teicoplanin patients (P = 0.21). Posttherapy asymptomatic C. difficile carriage (positive follow-up cultures without any clinical symptoms) occurred in five (25%) vancomycin patients and two (7.7%) teicoplanin patients (P = 0.11).Overall, 9 of 20 (45%) vancomycin patients and 5 of 26 (19.2%) teicoplanin patients (P=0.059) appeared not to be cleared of C. difficile after treatment. No adverse effects related to vancomycin or teicoplanin therapy were observed.


Proposal of a classification system for patients with neck pain.
It is likely that patients with neck pain are not a homogeneous group, but, instead, consist of a variety of subgroups, each of which may benefit from a specific intervention matched to the patient's signs and symptoms. Studies to date have largely failed to account for this possibility, which may compromise the statistical power of research and ultimately fail to provide guidance for clinical decision making. Classification provides a means of breaking down a larger entity into more homogeneous subgroups of patients, based on examination data. Classification can guide the determination of a patient's prognosis, and the selection of the most appropriate intervention strategy. Classification has received considerable attention in the management of patients with low back pain, and evidence is emerging regarding its benefits. There has been considerably less effort made towards examining classification as it pertains to patients with neck pain. The purpose of this clinical commentary is to examine the current literature and to propose a classification system for patients with neck pain, based on the overall goal of treatment. The approach is based on published evidence when possible and is also informed by clinical experience and expert opinion. Classification decisions are based on the integration of data from a variety of information from the history and physical examination. The end result of the classification process is to determine the treatment approach believed to be most likely to maximize the clinical outcome for an individual patient with neck pain.


Binary mixtures of (N-phosphonomethyl)-glycine with new aminophosphonates.
The potential biological activity of binary mixtures of some new organophosphorous compounds, aminoalkane- and aminofluorenephosphonates, with (N-phosphonomethyl)-glycine (glyphosate, PMG) was studied. The inhibition of growth of wheat (Triticum aestivum) induced by individual compounds and their equimolar mixtures with PMG was a measure of that activity. The experiments were expected to show if the new compounds exhibited good biological activity to be used for agrochemical applications and if this activity can be improved when they are used in mixtures with glyphosate which is the active component of the well-known herbicide Roundup. The results obtained show that aminofluorenephosphonates inhibited wheat growth when used in micromolar concentrations. Thus, their efficiency can be compared to that of PMG. The efficiency of aminoalkanephosphonates was one order of magnitude weaker. The measure of the efficiency was the effective concentration inhibiting wheat growth by 50% (EC50). The most demanded interaction, i.e., a synergistic was observed for only one of binary mixtures of the compounds studied with PMG. Mostly they showed antagonistic or strong antagonistic interactions. Some of them were of the additive type. Such results exclude the possibility of potential use of all the compounds studied in binary mixtures with phosphonomethylglycine, especially as the mentioned synergistic interaction found was rather weak. The influence of structural features of anminophosphonates on the results obtained is discussed.


Reversal of cyclosporine-induced mortality with a synthetic polymeric immunostimulant in a murine model of fecal peritonitis.
We have been investigating the effects of a synthetic immunostimulative polymer known as copovithane (Cpv). This agent appears to enhance humoral immunity in untreated and cyclosporine-immunosuppressed mice and is nontoxic in rodents and man. The purpose of this study was to determine whether cyclosporine (CsA) is deleterious to survival in a murine cecal ligation, puncture, and excision (CLPE) model of fecal peritonitis, and--if so--whether this effect could be ameliorated by Cpv without interfering with skin allograft acceptance. Cpv significantly prolongs survival in the CLPE model; the optimal dose for this effect was found to be 100 mg/kg. CsA was found to have a significant and deleterious effect on survival at several dosage levels when administrated 48 and 24 hr before cecal ligation, and immediately before and 16 hr after cecal ligation. Using a dose of CsA sufficient for skin allograft acceptance and the same schedule of administration outlined above, Cpv 100 mg/kg was administered 48 hr prior to cecal ligation. Mice treated with CsA plus Cpv had significantly longer survival than mice treated with CsA alone; furthermore, the survival of CsA-plus-Cpv-treated animals was not significantly different from that of saline-treated controls. Acceptance and survival of H-2 incompatible skin allografts in mice treated with CsA were not affected by Cpv 100 mg/kg/week. We conclude that CsA-induced mortality in the CLPE model can be abrogated by Cpv without adversely affecting skin allograft survival. It may eventually be possible to reduce the incidence of septic complications in clinical allotransplantation by prophylactically administering Cpv to patients on CsA immunosuppression.


A Huge Subcutaneous Hematoma in an Adult with Kasabach-Merritt Syndrome.
BACKGROUND Kasabach-Merritt syndrome is a potentially fatal disease that consists of hemangioma(s) with thrombocytopenia, microangiopathic hemolytic anemia, and coagulopathy. Extensive hemangiomatosis is rare. We present the radiological features and treatment strategy of a young adult suffering from Kasabach-Merritt syndrome with widespread hemangiomas and an infected huge hematoma in the right thigh. CASE REPORT A 33-year-old Taiwanese male presented with a painful 20-cm mass over his right thigh and gross hematuria for 2 days. Hemangiomatosis was bioptically proven in infancy and the patient was under regular follow-up. Physical examination revealed normal heart rate, respiratory rate, and body temperature. Multiple palpable lumps with brown and purple areas of skin over the neck, trunk, and right thigh were noted. Laboratory examinations revealed thrombocytopenia anemia and elevated fibrin degradation products. There were no signs of sepsis. Blood transfusion and steroid therapy were executed. Computed tomography showed a huge complicated subcutaneous hematoma in the right thigh. Drainage of the huge hematoma was performed and antibiotics were prescribed. After the local infection in the right thigh and the bleeding tendency were controlled, the patient was discharged in a stable condition two weeks later. CONCLUSIONS A huge infected hematoma and widespread hemangiomas are extremely rare complications of Kasabach-Merritt syndrome. There are no known treatment guidelines currently available. Our patient was successfully treated with steroids, drainage, and antibiotics.


Haploinsufficiency for p190B RhoGAP inhibits MMTV-Neu tumor progression.
Rho signaling regulates key cellular processes including proliferation, survival, and migration, and it has been implicated in the development of many types of cancer including breast cancer. P190B Rho GTPase activating protein (RhoGAP) functions as a major inhibitor of the Rho GTPases. P190B is required for mammary gland morphogenesis, and overexpression of p190B in the mammary gland induces hyperplastic lesions. Hence, we hypothesized that p190B may play a pivotal role in mammary tumorigenesis. To investigate the effects of loss of p190B function on mammary tumor progression, p190B heterozygous mice were crossed with an MMTV-Neu breast cancer model. Effects of p190B deficiency on tumor latency, multiplicity, growth, preneoplastic progression and metastasis were evaluated. To investigate potential differences in tumor angiogenesis between the two groups, immunohistochemistry to detect von Willebrand factor was performed and quantified. To examine gene expression of potential mediators of the angiogenic switch, an angiogenesis PCR array was utilized and results were confirmed using immunohistochemistry. Finally, reciprocal transplantation of tumor fragments was performed to determine the impact of stromal deficiency of p190B on tumor angiogenesis. P190B deficiency reduced tumor penetrance (53% of p190B+/-Neu mice vs. 100% of p190B+/+Neu mice formed tumors) and markedly delayed tumor onset by an average of 46 weeks. Tumor multiplicity was also decreased, but an increase in the number of preneoplastic lesions was detected indicating that p190B deficiency inhibited preneoplastic progression. Angiogenesis was decreased in the p190B heterozygous tumors, and expression of a potent angiogenic inhibitor, thrombospondin-1, was elevated in p190B+/-Neu mammary glands. Transplantation of p190B+/-Neu tumor fragments into wild-type recipients restored tumor angiogenesis. Strikingly, p190B+/+Neu tumor fragments were unable to grow when transplanted into p190B+/-Neu recipients. These data suggest that p190B haploinsufficiency in the epithelium inhibits MMTV-Neu tumor initiation. Furthermore, p190B deficiency in the vasculature is responsible, in part, for the inhibition of MMTV-Neu tumor progression.


Invasive candidiasis and candidemia: from current opinions to future perspectives.
Candida spp. are the fourth most common cause of nosocomial bloodstream infections in the United States, as well as the single most important cause of opportunistic fungal infections worldwide. A delayed diagnosis of invasive candidiasis and/or inadequate treatment choice is associated with high mortality rates and prolonged hospital stays. Even though the antifungal armamentarium has been broadened significantly over the last years, the best options for diagnosing and treating invasive candidiasis still remain a matter of discussion. In this article we present and analyze current evidence on the epidemiology, diagnostic methods and treatment options of invasive candidiasis, with a focus on results from randomized clinical trials. Finally, the reader is provided with a brief overview on promising clinical trial designs and antifungals that might shape the treatment of invasive candidiasis in the years to come.


Biomechanical evaluation of malleable noncompression miniplates in mandibular angle fractures: an experimental study.
The purpose of this experimental study was to test the reliability of a single malleable titanium miniplate using Champy's method of fixing fractures of the mandibular angle. Eighteen sheep hemimandibles were used to evaluate 2 plating techniques. The groups were tested with either a single non-compression titanium miniplate or a single malleable titanium miniplate. A cantilever bending biomechanical test model was used for the samples. Each group was tested with vertical forces using a servohydraulic testing unit. The displacement values in each group at each 10N stage up to 90N were compared using 2-way analysis of variance (ANOVA). The displacement values for the 2 groups differed significantly (p<0.01). The variance analyses showed that the biomechanical behaviour of a single non-compression miniplate was better than that of a single malleable miniplate. The non-compression miniplate fixed by screws had greater resistance to occlusal loads than the malleable plate fixed by screws, and the malleable plate alone was not sufficient to withstand the early postoperative bite force.


[Molecular biology of brain meningiomas].
Meningiomas are by far the most common tumors arising from the minges. A myriad of aberrant signaling pathways involved with meningioma tumorigenesis, have been discovered. Understanding these disrupted pathways will aid in deciphering the relationship between various genetic changes and their downstream effects on meningioma pathogenesis. An understanding of the genetic and molecular profile of meningioma would provide a valuable first step towards developing more effective treatment for this intracranial tumor. Chromosomes 1, 10, 14, 22, their associated genes, have been linked to meningioma proliferation and progression. It is presumed that through an understanding of these genetic factors, more educated meningioma treatment techniques can be implemented. Future therapies will include combinations of targeted molecular agents including gene therapy, si-RNA mediation, proton therapy, and other approaches as a result of continued progress in the understanding of genetic and biological changes associated with meningiomas.


[Breast cancer screening: is family medicine intervention effective?].
A non-randomized controlled trial was conducted in two family medicine centers. The objectives were to evaluate whether or not a systematic prescription by family physicians of a screening mammography to women aged 50 to 69 belonging in majority to a disadvantaged socio-economic group, would permit to reach at least 60% of them and to explore which factors were associated with compliance to the prescription. The experimental intervention consisted in the prescription by the family physician of a mammogram to those patients found eligible for the screening procedure irrespective of the reason for encounter. A total of 468 of the 870 women who consulted a physician during the study period were eligible for a screening mammography. In the experimental group, the mammography prescription rate was 89% for eligible women. At the end of the study, 58.8% (95% Conf. Int.:51.9%-65.7%) of the women in the experimental group and 13.4% (95% Conf. Int.:9.4%-17.4%) of those in the control group had passed a mammography (p < 0.0001). The adjusted odds ratio for the family physician's intervention was 14.98 (95% Conf. Int.:7.86-28.58). Women over 60 years of age and those who perceived themself more vulnerable to breast cancer were more likely to respond positively to their physician's recommendation. This study confirms that family physicians can effectively reach, in the area of clinical prevention, individuals who might not be reached otherwise.


Relative abundance and localization of interferon-stimulated gene 15 mRNA transcript in intra- and extra-uterine tissues during the early stages of pregnancy in sheep.
The aim of this study was to investigate relative abundance and localization of ISG15 mRNA transcript in intra-uterine (trophoblast, endometrium) and extra-uterine (hypothalamus, anterior pituitary, corpus luteum) tissues before and during the period of conceptus implantation. Multiparous ewes (n = 16) were randomly allotted into four groups: pregnant or estrous cyclic on days of 12 and 16 (n = 4 per group) following estrus. Relative abundances of ISG15 mRNA transcript were determined in the endometrium, corpus luteum, hypothalamus, and anterior-pituitary using real time quantitative PCR. Localization of ISG15 mRNA transcript was evaluated using in situ hybridization. The presence of ISG15 mRNA transcript was only visualized in intra-uterine tissues including the endometrium and trophoblast on day 12 of pregnancy. The ISG15 mRNA transcript was detected in all tissues evaluated on day 16 of pregnancy. The abundance of ISG15 mRNA transcript was greater in the endometrium on day 12 of pregnancy than at other days when evaluations occurred while in all other tissues except the hypothalamus there were large abundances of ISG15 mRNA on day 16 of pregnancy. It is concluded that the ISG15 mRNA transcript is only present in intra-uterine tissues before conceptus implantation. The ISG15 mRNA transcript, however, is present in extra-uterine tissues of ewes during implantation probably due to an increased amount of interferon-tau in blood circulation that is produced by the developing embryo. Results also indicate, for the first time, that pregnancy is associated with an intra-hypothalamus and anterior pituitary increased abundance of ISG15 mRNA transcript in ewes.


Monoclonal antibodies against the aster yellows agent.
Hybridoma clones secreting specific monoclonal antibodies against the aster yellows agent, a mycoplasma-like organism, were produced by using partially purified salivary gland preparations from infected leafhopper vectors as the immunogen. After 3947 hybridomas from 20 independent fusions were screened for specific antibody against the aster yellows agent, two table clones were obtained. With these monoclonal antibodies the aster yellows agent in diseased lettuce, periwinkles, and inoculative insects was specifically identified by enzyme-linked immunosorbent assay. The aster yellows agent was serologically differentiated from the mycoplasma-like organisms associated with ash yellows, loofah witches'-broom, paulownia witches'-broom, sweet potato witches'-broom, peanut rosette, maize bushy stunt, and elm phloem necrosis.


MiR-93 inhibition ameliorates OGD/R induced cardiomyocyte apoptosis by targeting Nrf2.
To identify the expression changes of microRNA 93 (miR-93) in oxygen-glucose deprivation/reoxygenation (OGD/R) injury in cardiomyocytes and its mechanism of mediating OGD/R and inducing apoptosis. Primary cardiomyocytes were extracted and OGD/R model in cardiomyocytes was established in vitro. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expressions of miR-93, and Western blot assay was applied to measure the protein levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and caspase-3. Flow cytometry was utilized to examine the cardiomyocyte apoptosis rate. The apoptosis rate was increased after OGD/R in cardiomyocytes, accompanied by remarkable rise of miR-93 expression. After transfection of miR-93 antagomir, the apoptosis rate of cardiomyocyte induced by OGD/R was down-regulated, and the expression of cleaved caspase-3 was decreased. Meanwhile, the results of qRT-PCR and Western blot showed that the levels of Nrf2 mRNA and protein expression were up-regulated after the miR-93 level was inhibited, and luciferase reporter assay affirmed that Nrf2 was a target molecule for OGD/R-induced apoptosis mediated by miR-93. miR-93 mediates OGD/R-induced hypoxia/reoxygenation injury apoptosis in cells by targeting Nrf2.


The power of mouse genetics to study spermatogenesis.
Approximately 80 million people worldwide are infertile, and nearly half of all infertility cases are attributed to a male factor. Therefore, progress in reproductive genetics becomes crucial for future diagnosis and treatment of infertility. In recent years, enormous progress has been made in this field. More than 400 mutant mouse models with specific reproductive abnormalities have been produced, and numerous human association studies have been discovered. However, the translation of basic science findings to clinical practice remains protracted, with only modest progress in the application of novel findings to clinical genetic testing and cures. To date, the most significant findings in male infertility remain numeric and structural chromosomal abnormalities and Y-chromosome microdeletions in infertile men. Thus, we anticipate that future genetic investigations will focus on infertile men with a normal somatic karyotype but with various spermatozoal defects, like insufficient production of spermatozoa (oligozoospermia), inadequate motility (asthenozoospermia), abnormal morphology (teratozoospermia), or combinations of these defects. Ultimately, basic advances in mammalian nonhuman reproduction will translate to clinical advances in human reproduction and testing for infertile humans, thereby helping to improve diagnostics and health care for infertile patients.


CeL-ID: cell line identification using RNA-seq data.
Cell lines form the cornerstone of cell-based experimentation studies into understanding the underlying mechanisms of normal and disease biology including cancer. However, it is commonly acknowledged that contamination of cell lines is a prevalent problem affecting biomedical science and available methods for cell line authentication suffer from limited access as well as being too daunting and time-consuming for many researchers. Therefore, a new and cost effective approach for authentication and quality control of cell lines is needed. We have developed a new RNA-seq based approach named CeL-ID for cell line authentication. CeL-ID uses RNA-seq data to identify variants and compare with variant profiles of other cell lines. RNA-seq data for 934 CCLE cell lines downloaded from NCI GDC were used to generate cell line specific variant profiles and pair-wise correlations were calculated using frequencies and depth of coverage values of all the variants. Comparative analysis of variant profiles revealed that variant profiles differ significantly from cell line to cell line whereas identical, synonymous and derivative cell lines share high variant identity and are highly correlated (ρ > 0.9). Our benchmarking studies revealed that CeL-ID method can identify a cell line with high accuracy and can be a valuable tool of cell line authentication in biomedical science. Finally, CeL-ID estimates the possible cross contamination using linear mixture model if no perfect match was detected. In this study, we show the utility of an RNA-seq based approach for cell line authentication. Our comparative analysis of variant profiles derived from RNA-seq data revealed that variant profiles of each cell line are distinct and overall share low variant identity with other cell lines whereas identical or synonymous cell lines show significantly high variant identity and hence variant profiles can be used as a discriminatory/identifying feature in cell authentication model.


Various members of the Toll-like receptor family contribute to the innate immune response of human epidermal keratinocytes.
Toll-like receptors (TLRs) are important pattern recognition molecules that activate the nuclear factor (NF)-kappaB pathway leading to the production of antimicrobial immune mediators. As keratinocytes represent the first barrier against exogenous pathogens in human skin, we investigated their complete functional TLR1-10 expression profile. First, reverse transcription-polymerase chain reaction (PCR) analysis revealed a very similar pattern of TLR mRNA expression when comparing freshly isolated human epidermis and cultured primary human keratinocytes. Thus, further experiments were carried out with primary keratinocytes in comparison with the spontaneously immortalized human keratinocyte cell line HaCaT. The quantitative expression of TLR1-10 mRNA in real-time PCR of primary human keratinocytes and HaCaT cells was analysed. Both cell types constitutively expressed TLR2, TLR3, TLR5, and to a lesser extent TLR10. TLR4 was only found in HaCaT cells, TLR1 to a higher degree in primary keratinocytes. In line with this, LPS induced mRNA expression of CD14 and TLR4 only in HaCaT cells. After stimulation with various TLR ligands, the NF-kappaB-activated chemokine interleukin-8 (IL-8) was measured. In primary keratinocytes and HaCaT cells the TLR3 ligand poly (I:C) was the most potent stimulator of IL-8 secretion. The TLR ligands peptidoglycan, Pam3Cys and flagellin which bind to TLR2, TLR1/TLR2 heterodimer, and TLR5, respectively, also induced IL-8 secretion, whereas no IL-8 was induced by LPS, R-848, loxoribine and cytosine guanine dinucleotide-containing oligodeoxynucleotide. A corresponding pattern was found in the RelA NF-kappaB translocation assay after ligand stimulation of primary keratinocytes. These studies provide substantial evidence for a functional TLR expression and signalling profile of normal human keratinocytes contributing to the antimicrobial defence barrier of human skin.


[Pulmonary ventilation and central hemodynamics in pulmonary tuberculosis patients].
Computerized devices were used for evaluation of the state of pulmonary ventilation and central hemodynamics in 32 patients with pulmonary tuberculosis depending on the spread of the specific process. Patients with limited changes in the lungs, showed disorders of bronchial patency and compensatory increase of organic ejection of the right ventricle. In the presence of spread changes in the lungs the above mentioned changes were supplemented by restrictive ventilation disorders with a reduction of the contractile myocardial capacity. These functional changes should be considered in the treatment of patients with pulmonary tuberculosis.


Analytical approaches to support current understanding of exposure, uptake and distributions of engineered nanoparticles by aquatic and terrestrial organisms.
Initiatives to support the sustainable development of the nanotechnology sector have led to rapid growth in research on the environmental fate, hazards and risk of engineered nanoparticles (ENP). As the field has matured over the last 10 years, a detailed picture of the best methods to track potential forms of exposure, their uptake routes and best methods to identify and track internal fate and distributions following assimilation into organisms has begun to emerge. Here we summarise the current state of the field, focussing particularly on metal and metal oxide ENPs. Studies to date have shown that ENPs undergo a range of physical and chemical transformations in the environment to the extent that exposures to pristine well dispersed materials will occur only rarely in nature. Methods to track assimilation and internal distributions must, therefore, be capable of detecting these modified forms. The uptake mechanisms involved in ENP assimilation may include a range of trans-cellular trafficking and distribution pathways, which can be followed by passage to intracellular compartments. To trace toxicokinetics and distributions, analytical and imaging approaches are available to determine rates, states and forms. When used hierarchically, these tools can map ENP distributions to specific target organs, cell types and organelles, such as endosomes, caveolae and lysosomes and assess speciation states. The first decade of ENP ecotoxicology research, thus, points to an emerging paradigm where exposure is to transformed materials transported into tissues and cells via passive and active pathways within which they can be assimilated and therein identified using a tiered analytical and imaging approach.


Relation of midwall circumferential systolic stress to equatorial midwall fibre shortening in chronic aortic regurgitation. Value as a predictor of postoperative outcome.
Nineteen patients with chronic aortic regurgitation and a large increase in heart size were studied before aortic valve replacement. By relating midwall circumferential systolic stress to midwall circumferential fibre shortening (Cs/Cd) before operation the patients could be divided into two well defined groups. Twelve patients (group 1) had a pronounced decrease in heart size as measured by the cardiothoracic ratio and an excellent clinical outcome six months after operation. Seven patients (group 2) had no significant decrease in heart size and a less good clinical outcome. The ratio of midwall circumferential systolic stress to end systolic volume index was significantly higher in group 1 than in group 2. Group 2 had more severe left ventricular hypertrophy determined by the ratio of the wall thickness to the minor internal radius of the left ventricle (h:r ratio), total left ventricular mass, and left ventricular mass to end diastolic volume ratio. There were no significant differences in any other haemodynamic or angiographic indices between the two groups. Thus the relation of midwall circumferential systolic stress to fibre shortening is useful in determining the prognosis in individual patients with chronic aortic regurgitation undergoing aortic valve replacement.


From birds to humans: new concepts on airways relative to alveolar surfactant.
Pulmonary surfactant is a surface-active mixture of phospholipids and specific proteins that lines the epithelial surfaces of mammalian lungs. In the alveoli, its main function is to reduce surface tension to ensure that these structures can remain open during respiratory cycles of contraction and expansion. However, surfactant is also present in the conducting airways, even though they are relatively rigid and do not need a system capable of rapidly lowering surface tension in response to compression. This has raised the question whether there is a difference in composition and function between airway and alveolar surfactant. Interest in this question has been stimulated further by the recognition that surfactant also has important functions in the immune defenses of the respiratory tract. In this review, we describe differences that have been reported between human airway and alveolar surfactant. In addition, we draw parallels between human airway surfactant and surfactant from the lungs of birds. The latter are tubular and rigid and do not undergo cycles of contraction and expansion, thus more resembling the human conducting airways than alveoli. Using this as a model, we propose a new hypothesis to explain structural and functional differences between human airway and alveolar surfactant. We suggest that the molecular composition of surfactant is adapted to differences in the architecture of pulmonary surfaces and to the dynamics of surface area changes during respiration.


Unmet Needs of Parents of Children with Urea Cycle Disorders.
(1) Background: Phenotypic diversity and long-term health outcomes of individuals with urea cycle disorders (UCDs) have been described in detail. However, there is limited information on the burden on affected families. (2) Methods: To evaluate the family burden in parents with children suffering from UCDs, we used validated questionnaires. Socio-demographic characteristics were evaluated, and an adapted version of the Parental Need Scale for Rare Diseases questionnaire was used. The survey was conducted in families of UCD patients cared for at the University Children's Hospital Heidelberg. (3) Results: From April to November 2021, 59 participants were interviewed (mothers n = 34, fathers n = 25). The affected patients most frequently suffered from ornithine transcarbamylase deficiency (OTC-D) (female n = 12, male n = 12), followed by argininosuccinate synthetase deficiency (ASS-D, n = 13) and argininosuccinate lyase deficiency (ASL-D, n = 8). About one-third of the participants were "dissatisfied" or "extremely dissatisfied" with health professionals' disease knowledge. In addition, 30% of the participants reported a medium or high need for "additional information on the development of their children", and 44% reported a medium or high need "for information on available services". A majority of 68% reported a need for additional support regarding services such as support groups (42%) or psychological counseling (29%). (4) Conclusions: Our study indicates that there is an unmet need for sufficient information about the development of children with UCDs, as well as for information about available support services for families with UCD patients. Furthermore, the results highlight the importance of establishing or improving family-centered care approaches. This pilot study may serve as a template for the assessment of the family burden associated with other inherited metabolic diseases.


The influence of uncertain map features on risk beliefs and perceived ambiguity for maps of modeled cancer risk from air pollution.
Maps are often used to convey information generated by models, for example, modeled cancer risk from air pollution. The concrete nature of images, such as maps, may convey more certainty than warranted for modeled information. Three map features were selected to communicate the uncertainty of modeled cancer risk: (i) map contours appeared in or out of focus, (ii) one or three colors were used, and (iii) a verbal-relative or numeric risk expression was used in the legend. Study aims were to assess how these features influenced risk beliefs and the ambiguity of risk beliefs at four assigned map locations that varied by risk level. We applied an integrated conceptual framework to conduct this full factorial experiment with 32 maps that varied by the three dichotomous features and four risk levels; 826 university students participated. Data was analyzed using structural equation modeling. Unfocused contours and the verbal-relative risk expression generated more ambiguity than their counterparts. Focused contours generated stronger risk beliefs for higher risk levels and weaker beliefs for lower risk levels. Number of colors had minimal influence. The magnitude of risk level, conveyed using incrementally darker shading, had a substantial dose-response influence on the strength of risk beliefs. Personal characteristics of prior beliefs and numeracy also had substantial influences. Bottom-up and top-down information processing suggest why iconic visual features of incremental shading and contour focus had the strongest visual influences on risk beliefs and ambiguity. Variations in contour focus and risk expression show promise for fostering appropriate levels of ambiguity.


Anatomical specificity of functional amygdala imaging of responses to stimuli with positive and negative emotional valence.
Non-invasive neuroimaging is increasingly used for investigating the human amygdala. Accurate functional localization in the amygdala region is, however, challenging and quantitative data on the anatomical specificity of functional amygdala imaging is lacking. We have therefore retrospectively investigated 114 recently published human functional imaging studies concerned with the amygdala. We determined the anatomical assignment probabilities of a total of 339 reported activation sites to the amygdala defined using a cytoarchitectonically verified probabilistic atlas system. We find that approximately 50% of reported responses were located in the region with high probability (> or =80%) of belonging to the amygdala. This group included responses related both to stimuli of positive and negative emotional valence. Approximately 10% of reported response sites were assigned to the hippocampus, with up to 100% assignment probability. The remaining peaks were either located in the border regions of the amygdala and/or hippocampus or outside of both of these structures. Within the amygdala, the majority of peaks (96.3%) were found in the laterobasal (LB) and superficial (SF) subregions. Only 3.7% of peaks were found in the centromedial group (CM), possibly because anatomically delineating the CM region of the amygdala is particularly difficult and hence its extent might have been underestimated. Moreover, these results show that a core region of the amygdala is responsive to stimuli both of positive and negative emotional valence. The current findings highlight the usefulness of probabilistic amygdala maps and also point to a need for the development of accurate in vivo delineation and parcellation of the amygdala.


An asymmetric fitting function for condensed-phase Raman spectroscopy.
Asymmetric lineshapes are experimentally observed in the Raman spectra of different classes of condensed matter. Determination of the peak parameters, typically done with symmetric pseudo-Voigt functions, in such situations yields unreliable results. While a wide choice of asymmetric fitting functions is possible, for the function to be practically useful, it should satisfy several criteria: simple analytic form, minimum parameters, description of the symmetric shape as "zero case", estimation of the desired peak parameters in a straightforward way and, above all, adequate description of the experimental data. In this work we formulate the asymmetric pseudo-Voigt function by damped perturbation of the original symmetric shapes with one asymmetry-related parameter. The damped character of the perturbation ensures by construction consistent behavior of the line tails. We test the asymmetric function by fitting the experimental Raman spectra. The results show that the function is able to describe a wide range of experimentally observed asymmetries for different natures of asymmetric broadening, including 3D and 2D crystals, nanoparticles, polymers, and molecular solids and liquids.


Diagnostic performance of reformatted isotropic thin-section helical CT images in the detection of superior semicircular canal dehiscence.
Purpose The purpose of this article is to assess the diagnostic performance of computed tomography (CT) reformatted images for detection of superior semicircular canal (SSC) dehiscence. Material and methods Forty-two patients, with sound- and/or pressure-induced vestibular symptoms, and 42 control participants underwent helical CT examination with a highly collimated beam (0.5 mm). Reformatted images of the vestibular labyrinth were obtained in the standard axial and coronal planes (group A images), and in a plane parallel and perpendicular to the SSC (group B images). Diagnostic performance obtained by evaluating the group A images alone and the group B images alone was analyzed by using the area under the receiver operating characteristic curve (AUC). Results The diagnostic performance of group A images was AUC = 0.929 with an overall accuracy of 92.9%. The diagnostic performance of group B images was AUC = 0.988 with an overall accuracy of 98.8%. The evaluation of group B images alone showed an improved diagnostic performance over the group A images alone. Conclusion Thin-section 0.5-mm collimation CT with reformatted images oriented in the plane parallel and perpendicular to the SSC improves diagnostic accuracy in assessing for SSC dehiscence in comparison to CT images with reconstructions limited to traditional axial and coronal planes.


Determination of the natural abundance delta15N of nicotine and related alkaloids by gas chromatography/isotope ratio mass spectrometry.
A method is described by which the natural abundance delta15N values of nicotine, analogues, and metabolites can be determined. The alkaloids are extracted from their biological matrix by solid-phase extraction and analysis is conducted using isotope ratio mass spectrometry interfaced to gas chromatography. Repeatability and precision are sufficient to allow differences in the delta15N values of less than 1.0 per thousand to be satisfactorily measured, with a standard deviation routinely less than 0.5 per thousand. The methodology has been tested by determining the changes in the delta15N values of nicotine, N-methyl-2-phenylpyrrolidine and their respective demethylation products, nornicotine and 2-phenylpyrrolidine, during biotransformation by cell suspension cultures of Nicotiana species. Sufficient precision and reproducibility were obtained to allow the kinetic isotope effects associated with the demethylation reaction to be calculated.


Integrating life history and cross-immunity into the evolutionary dynamics of pathogens.
Models for the diversity and evolution of pathogens have branched into two main directions: the adaptive dynamics of quantitative life-history traits (notably virulence) and the maintenance and invasion of multiple, antigenically diverse strains that interact with the host's immune memory. In a first attempt to reconcile these two approaches, we developed a simple modelling framework where two strains of pathogens, defined by a pair of life-history traits (infectious period and infectivity), interfere through a given level of cross-immunity. We used whooping cough as a potential example, but the framework proposed here could be applied to other acute infectious diseases. Specifically, we analysed the effects of these parameters on the invasion dynamics of one strain into a population, where the second strain is endemic. Whereas the deterministic version of the model converges towards stable coexistence of the two strains in most cases, stochastic simulations showed that transient epidemic dynamics can cause the extinction of either strain. Thus ecological dynamics, modulated by the immune parameters, eventually determine the adaptive value of different pathogen genotypes. We advocate an integrative view of pathogen dynamics at the crossroads of immunology, epidemiology and evolution, as a way towards efficient control of infectious diseases.


Oxytocin decreases sweet taste sensitivity in mice.
Oxytocin (OXT) suppresses food intake and lack of OXT leads to overconsumption of sucrose. Taste bud cells were recently discovered to express OXT-receptor. In the present study we tested whether administering OXT to wild-type mice affects their licking behavior for tastants in a paradigm designed to be sensitive to taste perception. We injected C57BL/6J mice intraperitoneally (i.p.) with 10mg/kg OXT and assayed their brief-access lick responses, motivated by water deprivation, to NaCl (300mM), citric acid (20mM), quinine (0.3mM), saccharin (10mM), and a mix of MSG and IMP (100mM and 0.5mM respectively). OXT had no effect on licking for NaCl, citric acid, or quinine. A possible effect of OXT on saccharin and MSG+IMP was difficult to interpret due to unexpectedly low lick rates to water (the vehicle for all taste solutions), likely caused by the use of a high OXT dose that suppressed licking and other behaviors. A subsequent experiment focused on another preferred tastant, sucrose, and employed a much lower OXT dose (0.1mg/kg). This modification, based on our measurements of plasma OXT following i.p. injection, permitted us to elevate plasma [OXT] sufficiently to preferentially activate taste bud cells. OXT at this low dose significantly reduced licking responses to 0.3M sucrose, and overall shifted the sucrose concentration - behavioral response curves rightward (mean EC50saline=0.362M vs. EC50OXT=0.466M). Males did not differ from females under any condition in this study. We propose that circulating oxytocin is another factor that modulates taste-based behavior.


EFNS-ENS Guidelines on the diagnosis and management of disorders associated with dementia.
The last version of the EFNS dementia guidelines is from 2007. In 2010, the revised guidelines for Alzheimer's disease (AD) were published. The current guidelines involve the revision of the dementia syndromes outside of AD, notably vascular cognitive impairment, frontotemporal lobar degeneration, dementia with Lewy bodies, corticobasal syndrome, progressive supranuclear palsy, Parkinson's disease dementia, Huntington's disease, prion diseases, normal-pressure hydrocephalus, limbic encephalitis and other toxic and metabolic disorders. The aim is to present a peer-reviewed evidence-based statement for the guidance of practice for clinical neurologists, geriatricians, psychiatrists and other specialist physicians responsible for the care of patients with dementing disorders. It represents a statement of minimum desirable standards for practice guidance. The task force working group reviewed evidence from original research articles, meta-analyses and systematic reviews, published by June 2011. The evidence was classified (I, II, III, IV) and consensus recommendations graded (A, B, or C) according to the EFNS guidance. Where there was a lack of evidence, but clear consensus, good practice points were provided. New recommendations and good practice points are made for clinical diagnosis, blood tests, neuropsychology, neuroimaging, electroencephalography, cerebrospinal fluid (CSF) analysis, genetic testing, disclosure of diagnosis, treatment of behavioural and psychological symptoms in dementia, legal issues, counselling and support for caregivers. All recommendations were revised as compared with the previous EFNS guidelines. The specialist neurologist together with primary care physicians play an important role in the assessment, interpretation and treatment of symptoms, disability and needs of dementia patients.


Characterization of the passive responses of live skeletal muscle using the quasi-linear theory of viscoelasticity.
The tensile viscoelastic responses of live, innervated rabbit skeletal muscle were measured and characterized using the quasi-linear model of viscoelasticity. The tibialis anterior (TA) and extensor digitorum longus (EDL) muscles of anesthetized New Zealand white rabbits were surgically exposed and tested under in vivo conditions. Rate sensitivity of the force-time history was observed in response to constant velocity testing at rates from 0.01 to 2.0 Hz. Average hysteresis energy, expressed as a percentage of maximum stored strain energy, was 39.3 +/- 5.4% and was insensitive to deformation rate. The quasi-linear model, with constants derived from relaxation testing, was able to describe and predict these responses with correlation exceeding the 99% confidence interval for the 132 constant velocity tests performed (rmean = 0.9263 +/- 0.0373). The predictive ability of this model was improved when compressive loading effects on the muscle were neglected, rmean = 0.9306 +/- 0.0324. The rate insensitivity of hysteresis energy was predicted by the model; however, the absolute value of the hysteresis was underestimated (30.2 +/- 4.0%). Both muscles demonstrated strikingly different elastic functions. Geometric normalization of these responses (stress and strain) did not result in a single elastic function capable of describing both muscles. Based on these results, the quasi-linear model is recommended for the characterization of the structural responses of muscle; however, further investigation is required to determine the influence of muscle geometry and fiber architecture on the elastic function.


Characterizing the pocketome of Mycobacterium tuberculosis and application in rationalizing polypharmacological target selection.
Polypharmacology is beginning to emerge as an important concept in the field of drug discovery. However, there are no established approaches to either select appropriate target sets or design polypharmacological drugs. Here, we propose a structural-proteomics approach that utilizes the structural information of the binding sites at a genome-scale obtained through in-house algorithms to characterize the pocketome, yielding a list of ligands that can participate in various biochemical events in the mycobacterial cell. The pocket-type space is seen to be much larger than the sequence or fold-space, suggesting that variations at the site-level contribute significantly to functional repertoire of the organism. All-pair comparisons of binding sites within Mycobacterium tuberculosis (Mtb), pocket-similarity network construction and clustering result in identification of binding-site sets, each containing a group of similar binding sites, theoretically having a potential to interact with a common set of compounds. A polypharmacology index is formulated to rank targets by incorporating a measure of druggability and similarity to other pockets within the proteome. This study presents a rational approach to identify targets with polypharmacological potential along with possible drugs for repurposing, while simultaneously, obtaining clues on lead compounds for use in new drug-discovery pipelines.


[Infectious-inflammatory complications in patients after transplantation of the kidney].
The work deals with the results of retrospective analysis of infectious complications (IC) in 323 patients with terminal renal failure (TRF) who underwent 371 operations for allotransplantation of cadaver kidney (ATCK). IC frequency was 41%, their mortality, 42.6%. The structure of the IC was as follows: pulmonary 29.6%, wound 22.4%, urinary 20.4%, sepsis 19.1%, others 8.5%. Gram-negative microorganisms predominated in the etiology of the IC; gram-positive cocci, microbial associations, yeastlike fungi, and cytomegalovirus were encountered less frequently. The development of IC did not depend on the patients' sex, primary disease of the kidneys which led to TRF, or the duration of previous treatment by hemodialysis, but was directly related to the age of the recipients. It was found that the character of the basic and anticrisis immunosuppressive therapy influenced the frequency of IC occurrence. The authors recommend some preventive and therapeutic measures for reducing the risk of the development of IC in patients after ATCK.


Does Pain Medication Use Influence the Outcome of 8 Weeks of Education and Exercise Therapy in Patients with Knee or Hip Osteoarthritis? An Observational Study.
Patients with osteoarthritis are mainly managed in primary care settings and many patients use pain medication as symptomatic treatment. We investigated in OA-patients receiving an education and exercise program, the use and type of pain medication and its impact on outcomes at 3 months follow-up. The design was a retrospective cohort study using prospectively collected data from the GLA: D® registry. The study included 15,918 primary care patients. Among the included patients, 62% were pain medication users and 38% were non-users. Among the pain medications users, 35% were classified as paracetamol users, 54% as NSAID users, and 11% as opioid users. Medication users and non-users differed regarding a higher pain intensity, poorer physical and mental health. Pain medication use before and during the education and exercise program was associated with the pain intensity at 3 months follow-up. However, patients either using or not using pain medications improved over time, and the magnitude of the difference between patient groups was small (less than 10 mm on a 0-100 scale). Pain medication use is weakly associated with outcome at 3 months follow up in OA-patients receiving an education and exercise program. Between-group differences, however, are small and probably not clinically important.


Structure and function of a custom anticancer peptide, CB1a.
Several natural antimicrobial peptides including cecropins, magainins and melittins have been found to kill cancer cells. However, their efficacy may not be adequate for their development as anticancer agents. In this study, we used a natural antimicrobial peptide, cecropin B (CB), as a template to generate a novel anticancer peptide. Cecropin B is an amphipathic and polycationic peptide derived from the hemolymph of Hyalophora cecropia with well-known antimicrobial and cytolytic properties. The signature pattern of cecropins is W-x-(0,2)-[KDN]-x-{L}-K-[KRE]-[LI]-E-[RKN] (PROSITE: PS00268), and this signature sequence is located at N-terminus of CB. CB1a was constructed by repeating the N-terminal ten amino acids of CB three times and including a hinge near C-terminus. The circular dichroism spectra showed that CB1a is unstructured in aqueous solution, but adopt a helical conformation in membrane-like environment. The solution structure of CB1a in a polar solvent was also studied by NMR. CB1a formed a helix-hinge-helix in 20% HFIP solution, and it was found the bent angle between two helical segments was induced ranging from 60 degrees to 110 degrees . A heparin-binding motif is located in the central part of helix 1. Isothermal titration calorimetry reveals the association constant of CB1a bound to low molecular weight heparin is 1.66 x 10(5)M(-1) at physiological ionic strength at 25 degrees C. Binding of CB1a to heparin produces a large conformational change toward a more structural state. CB1a demonstrated promising activity against several cancer cells but low toxicity against non-cancer cells. The IC(50) of CB1a on leukemia and stomach carcinoma cells were in the range of 2-8-fold lower than those of CB. Besides, CB1a exhibited low hemolytic activity against human red blood cells. Due to these properties, CB1a has the potential to become a promising anticancer agent.


Miscategorized subset-knowers: Five- and six-knowers can compare only the numbers they know.
Initial acquisition of the first symbolic numbers is measured with the Give a Number (GaN) task. According to the classic method, it is assumed that children who know only 1, 2, 3, or 4 in the GaN task, (termed separately one-, two-, three-, and four-knowers, or collectively subset-knowers) have only a limited conceptual understanding of numbers. On the other hand, it is assumed that children who know larger numbers understand the fundamental properties of numbers (termed cardinality-principle-knowers), even if they do not know all the numbers as measured with the GaN task, that are in their counting list (e.g., five- or six-knowers). We argue that this practice may not be well-established. To validate this categorization method, here, the performances of groups with different GaN performances were measured separately in a symbolic comparison task. It was found that similar to one to four-knowers, five-, six-, and so forth, knowers can compare only the numbers that they know in the GaN task. We conclude that five-, six-, and so forth, knowers are subset-knowers because their conceptual understanding of numbers is fundamentally limited. We argue that knowledge of the cardinality principle should be identified with stricter criteria compared to the current practice in the literature. RESEARCH HIGHLIGHTS: Children who know numbers larger than 4 in the Give a Number (GaN) task are usually assumed to have a fundamental conceptual understanding of numbers. We tested children who know numbers larger than 4 but who do not know all the numbers in their counting list to see whether they compare numbers more similar to children who know only small numbers in the GaN task or to children who have more firm number knowledge. Five-, six-, and so forth, knowers can compare only the numbers they know in the GaN task, similar to the performance of the one, two, three, and four-knowers. We argue that these children have a limited conceptual understanding of numbers and that previous works may have miscategorized them.


Disease resistance is related to inherent swimming performance in Atlantic salmon.
Like humans, fish can be classified according to their athletic performance. Sustained exercise training of fish can improve growth and physical capacity, and recent results have documented improved disease resistance in exercised Atlantic salmon. In this study we investigated the effects of inherent swimming performance and exercise training on disease resistance in Atlantic salmon.Atlantic salmon were first classified as either poor or good according to their swimming performance in a screening test and then exercise trained for 10 weeks using one of two constant-velocity or two interval-velocity training regimes for comparison against control trained fish (low speed continuously). Disease resistance was assessed by a viral disease challenge test (infectious pancreatic necrosis) and gene expression analyses of the host response in selected organs. An inherently good swimming performance was associated with improved disease resistance, as good swimmers showed significantly better survival compared to poor swimmers in the viral challenge test. Differences in mortalities between poor and good swimmers were correlated with cardiac mRNA expression of virus responsive genes reflecting the infection status. Although not significant, fish trained at constant-velocity showed a trend towards higher survival than fish trained at either short or long intervals. Finally, only constant training at high intensity had a significant positive effect on fish growth compared to control trained fish. This is the first evidence suggesting that inherent swimming performance is associated with disease resistance in fish.


Trends in Tobacco Production and Prices in Malawi.
Diversification away from tobacco production has been identified as a priority for Malawi, historically one of the world's most tobacco-reliant countries. This paper refers to a nationally representative data set to characterize broad trends in production since 2000 to understand whether Malawi is shifting away from tobacco and how production has changed over time. From 2004 to 2019, the share of Malawian crop farmers producing tobacco fell from 16% to 5%, and tobacco's share of the total value of crop production also declined sharply. Tobacco farms are generally growing larger (in size and scale of production) over time. However, land productivity remains low, with net returns of 93 000 MWK (128 USD) per hectare at the median. Farm-gate prices for tobacco have declined relative to the prices of maize or fertilizer, rendering tobacco a less lucrative avenue to generate the cash income needed to purchase these key items. In addition, the share of the export price received by farmers has also declined over time, with the median farm-gate price representing 32% of the export price in 2004 and 18% in 2019. In some respects, a transition away from tobacco has already occurred. Additional research is needed to understand why the farm-gate share of tobacco export prices has declined over this period and how the livelihoods of smallholder farm-households that exited tobacco production have been affected. To the extent that tobacco prices appear to be declining, there is a need to rigorously assess whether farmers have suitable crop alternatives (with established markets) and other livelihood options. Likely, investments are yet needed to foster strong alternatives to tobacco; such investments include research and development in on-farm technologies to raise the productivity of non-tobacco crops, as well as improvements in non-tobacco value chains to reduce transportation costs and promote private investment.


Assessing Extracellular Vesicles in Human Biofluids Using Flow-Based Analyzers.
Extracellular vesicles (EVs) are increasingly being analyzed by flow cytometry. Yet their minuscule size and low refractive index cause the scatter intensity of most EVs to fall below the detection limit of most flow cytometers. A new class of devices, known as spectral flow analyzers, are becoming standards in cell phenotyping studies, largely due to their unique capacity to detect a vast panel of markers with higher sensitivity for light scatter detection. Another class of devices, known as nano-analyzers, provides high-resolution detection of sub-micron-sized particles. Here, the EV phenotyping performance between the Aurora (Cytek) spectral cell analyzer and the NanoFCM (nFCM) nanoflow analyzer are compared. These two devices are specifically chosen given their lead in becoming gold standards in their respective fields. Immune cell-derived EVs remain poorly characterized despite their clinical potential. Therefore, B- and T-cell line-derived EVs and donor-matched human biofluid-derived EVs from plasma, urine, and saliva are used in combination with a panel of established immune markers for this comparative study. A comparative evaluation of both cytometry platforms is performed, discussing their potential and suitability for different applications. It is found that nFCM can accurately i) analyze small EVs (40-200 nm) matching the size accuracy of electron microscopy; ii) measure the concentration of a single EV particle per volume; iii) identify underrepresented EV marker subsets; and iv) provide co-localization of EV surface markers. It can also be shown that human sample biofluids have unique EV marker signatures that can have future clinical relevance. Finally, nFCM and Aurora have their unique strength, preferred fashion of data acquisition, and visualization to fit different research interests.


The subcellular distribution of alanine-glyoxylate aminotransferase and serine-pyruvate aminotransferase in dog liver.
The subcellular distributions of alanine-glyoxylate aminotransferase and serine-pyruvate aminotransferase in the particulate fraction of dog liver were examined by centrifugation in a sucrose density gradient. Most of both enzyme activities in the particulate fraction were localized in the mitochondria, but not in the peroxisomes.


Fecal calprotectin as a biomarker of intestinal graft versus host disease after allogeneic hematopoietic stem cell transplantation.
The diagnosis of gastrointestinal graft versus host disease (GI-GVHD) is based on clinical symptoms and histological findings. In clinical practice, it is often difficult to decide whether abdominal symptoms in an allogeneic transplant recipient are caused by GVHD or other disorders. Endoscopic biopsies are helpful in establishing the diagnosis, but endoscopy is not always possible to perform due to poor general condition of the patients. No biomarkers are routinely used to predict GVHD. The aim of fecal calprotectin and alpha-1 antitrypsin testing in our study was to find out whether determination of the concentrations of these proteins may be used as a screening method for enteric GVHD. We studied prospectively 51 patients, 8 of whom developed GI-GVHD. Our data demonstrate that elevated fecal calprotectin levels were significantly associated with presence of GI-GVHD. We found a positive association between high F-calprotectin and severe gastrointestinal GVHD. In bivariate analysis, only calprotectin but not alpha-1 antitrypsin was independently associated with GI-GVHD. Testing for fecal calprotectin after allogeneic stem cell transplantation may be a useful screening tool.


Cortical lateralization patterns related to self-estimation of emotional state.
The relationships between subjectively-reported emotional state and hemispheric laterality were investigated. Participants' emotional state was modified using emotional slides. Self-estimation of Energy Arousal and Hedonic Tone (positive valence) as well as Tense Arousal (negative valence) was derived from the Activation-Deactivation Adjective Checklist and the UWIST Mood Adjective Checklist. Energy arousal was found to be associated with right frontal dominance in the alpha2 (10-12 Hz) band, together with left frontal dominance in the beta2 (16-24 Hz) band. It was also related to left alpha2 dominance in the central and centro-parietal cortex. The effects for the Hedonic Tone scale were limited to a frontal beta2 effect. Surprisingly, no effects of state estimates from the tension scales were observed. It can be concluded that selected qualities of subjective emotional state measured by adjective lists can be related in specific ways to hemispheric laterality, as measured by EEG methods.


Imputation of Missing Data for Time-to-Event Endpoints Using Retrieved Dropouts.
We have explored several statistical approaches to impute missing time-to-event data that arise from outcome trials with relatively long follow-up periods. Aligning with the primary estimand, such analyses evaluate the robustness of results by imposing an assumption different from censoring at random (CAR). Although there have been debates over which assumption and which method is more appropriate to be applied to the imputation, we propose to use the collection of retrieved dropouts as the basis of missing data imputation. As retrieved dropouts share a similar disposition, such as treatment discontinuation, with subjects who have missing data, they can reasonably be assumed to characterize the distribution of time-to-event among subjects with missing data. In terms of computational intensity and robustness to violation of underlying distributional assumption, we have compared parametric approaches via MCMC or MLE multivariate sampling procedures to a non-parametric bootstrap approach with respect to baseline hazard function. Each of these approaches follows a process of multiple imputation ("proper imputations"), analysis of complete datasets, and final combination. The type-I error, and power rates are examined under a wide range of scenarios to inform the performance characteristics. A subset of a real unblinded phase III CVOT is used to demonstrate the application of the proposed approaches, compared to the Cox proportional hazards model and jump-to-reference multiple imputation.


The Impact of Sepsis on the Outcomes of COPD Patients: A Population-Based Cohort Study.
This study aims to identify the impact of new-onset sepsis in patients with chronic obstructive pulmonary disease (COPD) including the effects on acute exacerbations, pneumonia and mortality. Using the National Health Insurance Research Database of Taiwan, all patients with COPD older than 40 years between 1988 and 2010 were recruited. After propensity score matching, each of the 8774 COPD patients with and without sepsis were identified to have similar characteristics. The primary outcome was severe exacerbations of COPD, with a severe exacerbation being defined as a patient requiring hospital admission or an emergency department visit due to COPD. The secondary outcomes were pneumonia, serious pneumonia, and all-cause mortality. The post-index overall cumulative incidence rates of total acute exacerbations were 11.2/person-years in the sepsis group and 6.2/person-years in the non-sepsis group (adjusted hazard ratio (HR) = 1.38, 95% confidence interval (CI), 1.38⁻1.40). The sepsis group also had higher risks of severe exacerbations (adjusted HR = 2.05, 95% CI, 2.02⁻2.08), severe exacerbations requiring hospitalization (adjusted HR = 2.30, 95% CI, 2.24⁻2.36), and severe exacerbations leading to an emergency room visit (adjusted HR = 1.91, 95% CI, 1.87⁻1.94). Regarding the effect on secondary outcomes, the sepsis group had higher risks of mortality (incidence rate: 23.7/person-years vs. 11.34/person-years, adjusted HR = 2.27, 95% CI, 2.14⁻2.41), pneumonia (incidence rate: 26.41 per person-days vs. 10.34 per person-days, adjusted HR = 2.70, 95% CI, 2.5⁻2.91), and serious pneumonia (incidence rate: 5.84 per person-days vs. 1.98 per person-days, adjusted HR = 2.89, 95% CI, 2.5⁻3.33) compared with the non-sepsis group. Sepsis survivors among patients with COPD had a higher risk of severe exacerbations, pneumonia, serious pneumonia, and mortality compared to patients with COPD without sepsis.


A ring of negative charges in the intracellular vestibule of Kir2.1 channel modulates K+ permeation.
The glutamate at site 224 of a Kir2.1 channel plays an important role in K+ permeation. The single-channel inward current flickers with reduced conductance in an E224G mutant. We show that open-channel fluctuations can also be observed in E224C, E224K, and E224Q mutants. Yet, open-channel fluctuations were not observed in either the wild-type or an E224D mutant. Introducing a negatively charged methanethiosulfonate reagent to the E224C mutant irreversibly increased channel conductance and eliminated open-channel fluctuations. These results suggest that although the negatively charged residue 224 is located at the internal vestibule, it is important for smooth inward K+ conduction. We identified a substate in the E224G mutant and showed that open-channel fluctuations are mainly attributed to rapid transitions between the substate and the main state. Also, we characterized the voltage- and ion-dependence of the substate kinetics. The open-channel fluctuations decreased in internal NH4+ or Tl+ as compared to internal K+. These results suggest that NH4+ and Tl+ gate the E224G mutant in a more stable state. Based on an ion-conduction model, we propose that the appearance of the substate in the E224G mutant is due to changes of ion gating in association with variations of ion-ion interaction in the permeation pathway.


Lymphocyte subpopulations and apoptosis of immune cells in rabbits experimentally infected with a strain of the RHD virus having a variable haemagglutination capacity.
The paper describes the immunological response in the matter of percentage of T cells (receptor CD5+) and subpopulations (Th with receptor CD4+, Tc/Ts with receptor CD8+, T with receptor CD25+) and B cells with receptor CD19+, as well as the percentage of apoptotic granulocytes and lymphocytes, in rabbits experimentally infected with the Hagenow strain of the RHD virus. The material chosen for the experiment is special, as among all strains of RHD virus, there are only two strains which carry the variable haemagglutination capacity of red cells. The results of the study show that the Hagenow strain gives an untypical picture of T and B lymphocytes, whereas the results in inducing apoptosis seems to corespond with previous data, confirming the inclusion of apoptosis from 4 h p.i. and the intensity of the phenomenon being higher in granulocytes.


Ambient Conditions Prior to Tokyo 2020 Olympic and Paralympic Games: Considerations for Acclimation or Acclimatization Strategies.
The Tokyo Olympics and Paralympic games in 2020 will be held in hot and humid conditions. Heat acclimation (in a climatic chamber) or heat acclimatization (natural environment) is essential to prepare the (endurance) athletes and reduce the performance loss associated with work in the heat. Based on the 1990-2018 hourly meteorological data of Tokyo and the derived wet bulb globe temperature (WBGT) (Liljegren method), Heat Index and Humidex, it is shown that the circumstances prior to the games are likely not sufficiently hot to fully adapt to the heat. For instance, the WBGT 2 weeks prior to the games at the hottest moment of the day (13:00 h) is 26.4 ± 2.9°C and 28.6 ± 2.8°C during the games. These values include correction for global warming. The daily variation in thermal strain indices during the Tokyo Olympics (WBGT varying by 4°C between the early morning and the early afternoon) implies that the time of day of the event has a considerable impact on heat strain. The Paralympics heat strain is about 1.5°C WBGT lower than the Olympics, but may still impose considerable heat strain since the Paralympic athletes often have a reduced ability to thermoregulate. It is therefore recommended to acclimate about 1 month prior to the Olympics under controlled conditions set to the worst-case Tokyo climate and re-acclimatize in Japan or surroundings just prior to the Olympics.


ZnO Nanoparticles from Different Precursors and Their Photocatalytic Potential for Biomedical Use.
Semiconductor photocatalysts, particularly ZnO nanoparticles, were synthesized via the precipitation method using four different precursors (zinc acetate/zinc nitrate/zinc sulfate/zinc chloride) and compared, according to their optical, structural, photocatalytic, and anticancer properties. The materials were characterized via X-ray Diffraction method (XRD), micro-Raman, Fourier Transform Infrared Spectroscopy (FT-IR), Brunauer-Emmett-Teller (BET), Dynamic Light Scattering (DLS), and Field Emission Scanning Electron Microscope (FESEM) analysis. Photocatalysis was conducted under UV and visible light irradiation, using Rhodamine B as the organic pollutant. It was observed that the highest photocatalysis efficiency was obtained by the nanoparticles synthesized from the zinc acetate used as precursor material. A cell-dependent anticancer efficiency of the tested ZnO nanoparticles was also observed, that was also attributed to the different precursors and the synthesis method, revealing that the nanoparticles that were synthesized from zinc acetate were more bioactive among the four tested precursors. Overall, the data revealed that both the enhanced photocatalytic and biological activity of ZnO nanoparticles derived from zinc acetate precursor could be attributed to the reduced crystalline size, increased surface area, as well as the observed hexagonal crystalline morphology.


Scientific Evidence Supporting Coronavirus Disease 2019 (COVID-19) Vaccine Efficacy and Safety in People Planning to Conceive or Who Are Pregnant or Lactating.
Three coronavirus disease 2019 (COVID-19) vaccines have been authorized for use in the United States; specifically, the Pfizer-BioNTech, Moderna, and Johnson & Johnson-Janssen COVID-19 vaccines were granted emergency use authorization by the U.S. Food and Drug Administration in late 2020 and early 2021. Vaccination coverage and intent among adults are lowest among those aged 18-39 years and among females in particular. In females of reproductive age, enthusiasm for receiving a COVID-19 vaccine may be negatively affected by claims currently circulating widely on diverse social media platforms regarding the vaccines adversely affecting fertility and pregnancy. Yet it is important to note that these claims are anecdotal in nature and not supported by the available scientific evidence. It is also imperative that the effects of COVID-19 vaccine on reproductive health are clarified. Herein, we discuss the existing scientific data supporting COVID-19 vaccine safety and efficacy in people who are planning to conceive or who are pregnant or lactating and highlight the importance of COVID-19 vaccination in females of reproductive age.


Functional proteomic analysis of Ankaferd® Blood Stopper.
Ankaferd® Blood Stopper (ABS) comprises a standardized mixture of the plants Thymus vulgaris, Glycyrrhiza glabra, Vitis vinifera, Alpinia officinarum, and Urtica dioica. The basic mechanism of action for ABS is the formation of an encapsulated protein network that provides focal points for vital erythrocyte aggregation. ABS-induced protein network formation with blood cells, particularly erythrocytes, covers the primary and secondary hemostatic system without disturbing individual coagulation factors. To understand the effect mechanisms of ABS on hemostasis, a proteomic analysis using 2D gel electrophoresis and mass spectrometer was performed. Proteins of plant origin in Ankaferd® were NADP-dependent-malic enzyme, ribulose bisphosphate-carboxylase-large chain, maturase K, ATP synthase subunit-beta, ATP synthase subunit-alpha, chalcone-flavanone isomerase-1, chalcone-flavanone isomerase-2, and actin-depolymerizing factor. Furthermore, functional proteomic studies revealed that proteins resembling human peptides have been detected within Ankaferd®, including ATP synthase, mucin-16 (CD164 sialomucin-like 2 protein), coiled-coil domain containing 141 hypothetical protein LOC283638 isoform 1, hypothetical protein LOC283638 isoform 2, dynactin 5, complex I intermediate-associated protein 30, mitochondrial, NADH dehydrogenase (ubiquinone) 1 alpha subcomplex, TP synthase, H+ transporting, mitochondrial actin binding 1 isoform, LIM domain and actin binding 1 isoform a, LIM domain and actin binding 1 isoform b, spectrin alpha non erythrocytic 1, prolactin releasing hormone receptor, utrophin, tet oncogene family member 2 isoform b, protein phosphatase 1 regulatory subunit 12A, NIMA (never in mitosis gene a)-related kinase, ATP-binding cassette protein C12, Homo sapiens malic enzyme 1, mitochondrial NADP(+)-dependent malic enzyme 3, ME2 protein, nuclear factor 1 B-type, abhydrolase domain-containing protein 12B, E3 SUMO-protein ligase PIAS2, alpha-1, 2-glucosyltransferase ALG10-A, cofilin, non-muscle isoform, 18 kDa phosphoprotein, p18, actin-depolymerizing factor (ADF), twinfilin-1, ankyrin repeat and FYVE domain-containing protein 1, usherin precursor, urotensin II receptor, interleukin 4, and midkine. Proteomic analysis of Ankaferd® represents a true basis for the upcoming Ankaferd® studies focusing on its wound healing, hemostatic, anti-infective, antineoplastic, and preservative biological actions.


Risk factors for heterotopic ossification in spinal cord injury.
Heterotopic ossification (HO) is a complication in 16% to 53% of spinal cord injured (SCI) patients. One third of these patients have moderate to severe HO that adversely affects function or health. Pharmacologic prophylaxis of HO for all SCI patients continues to be controversial. High-risk criteria for HO formation identified in total hip replacement patients are not applicable to SCI. A review of the literature did not reveal specific risk factors for HO with SCI. The charts of 100 randomly selected SCI patients, 50 with HO and 50 without HO, were reviewed retrospectively to learn if criteria which would predict high-risk patients could be identified. A total of 14 variables, seven demographic (age, sex, race, level of lesion, completeness of lesion, cause of injury, and geographic locus of patient) and seven medical (bladder stones, fractures, pressure sores, deep vein thrombosis, pulmonary embolism, spasticity, and urinary tract infections) were studied. Four of the 14 variables (age, completeness of lesion, presence of pressure sores, and spasticity) were significantly related to HO formation. The risk factors appear to be additive. When all were present, 92% of patients were found to have HO. Before the findings are applied clinically, it is suggested that a prospective study be conducted to confirm the risk predictive value of these factors in HO.


Acupuncture at Back-Shu and Front-Mu Acupoints Prevents Gastric Ulcer by Regulating the TLR4/MyD88/NF-κB Signaling Pathway.
To assess the preventive effects of acupuncture at back-shu and front-mu acupoints on rats with restraint water-immersion stress (RWIS)-induced gastric ulcer. Thirty-six rats were randomly divided into four groups for 10 days of treatment as follows: the normal group received no treatment; the model group received RWIS-induced gastric ulcer; the omeprazole group was administered omeprazole orally every 2 days; and the electroacupuncture group received electroacupuncture at the RN12 and BL21 acupoints every 2 days. After 10 days of treatment, except for the normal group, all rats were induced with gastric ulcer by RWIS for 3 h. The ulcer index (UI), ulcer inhibition rate, and histopathological score were calculated. We determined the levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in serum, and the activities of myeloperoxidase (MPO), malondialdehyde (MDA), superoxide dismutase (SOD), nitric oxide (NO), and glutathione peroxidase (GSH-Px) in serum and gastric tissues. Protein expression of MyD88, nuclear factor (NF)-κB (p65), and toll-like receptor (TLR) 4 was quantified in gastric tissues. The electroacupuncture and omeprazole groups were equivalent in terms of UI, ulcer inhibition rate, and histopathological score. The serum levels of TNF-α and IL-6 were significantly lower in the electroacupuncture group compared with the omeprazole group (P < 0.05). Compared with the model group, there were significant changes in the levels of NO, MPO, GSH-Px, and MDA in all other groups, while the expression of TLR4, MyD88, and NF-κB p65 in gastric tissue decreased significantly in the electroacupuncture group. The expression of TLR4 was substantially lower in the electroacupuncture group compared with the omeprazole group. Acupuncture at back-shu and front-mu acupoints played a role in preventing gastric ulcer by inhibiting extracellular signals, stimulating kinases in serum and gastric tissues, and activating the inhibition of the TLR4 signaling pathway.


Site-selective protonation of the one-electron reduced cofactor in [FeFe]-hydrogenase.
Hydrogenases are bidirectional redox enzymes that catalyze hydrogen turnover in archaea, bacteria, and algae. While all types of hydrogenase show H2 oxidation activity, [FeFe]-hydrogenases are excellent H2 evolution catalysts as well. Their active site cofactor comprises a [4Fe-4S] cluster covalently linked to a diiron site equipped with carbon monoxide and cyanide ligands. The active site niche is connected with the solvent by two distinct proton transfer pathways. To analyze the catalytic mechanism of [FeFe]-hydrogenase, we employ operando infrared spectroscopy and infrared spectro-electrochemistry. Titrating the pH under H2 oxidation or H2 evolution conditions reveals the influence of site-selective protonation on the equilibrium of reduced cofactor states. Governed by pKa differences across the active site niche and proton transfer pathways, we find that individual electrons are stabilized either at the [4Fe-4S] cluster (alkaline pH values) or at the diiron site (acidic pH values). This observation is discussed in the context of the complex interdependence of hydrogen turnover and bulk pH.


Adverse events associated with the use of cervical manipulation and mobilization for the treatment of neck pain in adults: a systematic review.
Adverse events (AE) are a concern for practitioners utilizing cervical manipulation or mobilization. While efficacious, these techniques are associated with rare but serious adverse events. Five bibliographic databases (PubMed, CINAHL, PEDro, AMED, EMBASE) and the gray literature were searched from 1998 to 2009 for any AE associated with cervical manipulation or mobilization for neck pain. Randomized controlled trials (RCTs), prospective or cross-sectional observational studies were included. Two independent reviewers conducted study selection, method quality assessment and data abstraction. Pooled relative risks (RR) were calculated. Study quality was assessed using the Cochrane system, a modified Critical Appraisal Skills Program form and the McHarm scale to assess the reporting of harms. Seventeen of 76 identified citations resulted in no major AE. Two pooled estimates for minor AE found transient neurological symptoms [RR 1.96 (95% CI: 1.09-3.54) p < 0.05]; and increased neck pain [RR 1.23 (95% CI: 0.85-1.77) p > .05]. Forty-four studies (58%) were excluded for not reporting AE. No definitive conclusions can be made due to a small number of studies, weak association, moderate study quality, and notable ascertainment bias. Improved reporting of AE in manual therapy trials as recommended by the CONSORT statement extension on harms reporting is warranted.


Smartphone-interfaced lab-on-a-chip devices for field-deployable enzyme-linked immunosorbent assay.
The emerging technologies on mobile-based diagnosis and bioanalytical detection have enabled powerful laboratory assays such as enzyme-linked immunosorbent assay (ELISA) to be conducted in field-use lab-on-a-chip devices. In this paper, we present a low-cost universal serial bus (USB)-interfaced mobile platform to perform microfluidic ELISA operations in detecting the presence and concentrations of BDE-47 (2,2',4,4'-tetrabromodiphenyl ether), an environmental contaminant found in our food supply with adverse health impact. Our point-of-care diagnostic device utilizes flexible interdigitated carbon black electrodes to convert electric current into a microfluidic pump via gas bubble expansion during electrolytic reaction. The micropump receives power from a mobile phone and transports BDE-47 analytes through the microfluidic device conducting competitive ELISA. Using variable domain of heavy chain antibodies (commonly referred to as single domain antibodies or Nanobodies), the proposed device is sensitive for a BDE-47 concentration range of 10(-3)-10(4 ) μg/l, with a comparable performance to that uses a standard competitive ELISA protocol. It is anticipated that the potential impact in mobile detection of health and environmental contaminants will prove beneficial to our community and low-resource environments.


Mapping the distribution of maize streak virus genotypes across the forest and transition zones of Ghana.
Throughout sub-Saharan Africa, maize streak virus strain A (MSV-A), the causal agent of maize streak disease (MSD), is an important biological constraint on maize production. In November/December 2010, an MSD survey was carried out in the forest and transition zones of Ghana in order to obtain MSV-A virulence sources for the development of MSD-resistant maize genotypes with agronomic properties suitable for these regions. In 79 well-distributed maize fields, the mean MSD incidence was 18.544 % and the symptom severity score was 2.956 (1 = no symptoms and 5 = extremely severe). We detected no correlation between these two variables. Phylogenetic analysis of cloned MSV-A isolates that were fully sequenced from samples collected in 51 of these fields, together with those sampled from various other parts of Africa, indicated that all of the Ghanaian isolates occurred within a broader cluster of West African isolates, all belonging to the highly virulent MSV-A1 subtype. Besides being the first report of a systematic MSV survey in Ghana, this study is the first to characterize the full-genome sequences of Ghanaian MSV isolates. The 51 genome sequences determined here will additionally be a valuable resource for the rational selection of representative MSV-A variant panels for MSD resistance screening.


HER2 and response to anthracycline-based neoadjuvant chemotherapy in breast cancer.
The predictive role of human epidermal growth factor receptor 2 (HER2) to adjuvant anthracycline-based chemotherapy remains controversial. Here, we investigated the association between HER2 status and pathological response in breast cancer patients who received neoadjuvant anthracycline-based regimens. Women (n = 538) with operable primary breast cancer received neoadjuvant anthracycline-based chemotherapy. Pathological complete response (pCR) was defined as no invasive breast tumor cells in breast after completion of neoadjuvant chemotherapy. HER2 status was determined by immunohistochemistry and/or by fluorescence in situ hybridization in core biopsy breast cancer tissue obtained before initiation of neoadjuvant chemotherapy. In this cohort of 538 patients, 23.9% of patients achieved a pCR in their breast. HER2-positive tumors had a lower rate of pCR than did HER2-negative tumors (14.7% versus 25.7%, P = 0.013); negative HER2 status remained as an independent favorable predictor of pCR after adjusted for age, estrogen receptor, progesterone receptor, tumor size, chemotherapy cycles, and tumor grade in a multivariate analysis (odds ratio = 3.14; 95% confidence interval = 1.60-6.16, P = 0.001). Furthermore, patients with a pCR had a higher 3-year disease-free survival (DFS) rate than did patients without a pCR (P = 0.007). Women with HER2-negative breast cancers rather than HER2-positive tumors benefit from anthracycline-based neoadjuvant chemotherapy.


Medicare Advantage in the era of health reform: progress in leveling the playing field.
Payments to private Medicare Advantage (MA) plans have exceeded Medicare fee-for-service (FFS) costs since those payments were increased by the Medicare Modernization Act of 2003 (MMA). Payments to MA plans in 2010 exceeded average costs in FFS Medicare nationally by 8.9 percent, a total of $8.9 billion. While these extra payments are substantial, they represent a decrease relative to 2009, when MA payments were 13.0 percent, or $11.4 billion, greater than FFS costs. The decrease in MA payments relative to FFS costs, while mostly resulting from policy decisions and other factors not directly related to the health reform law, begins to shift MA payments toward levels mandated in the provisions that are set to go into effect in 2012.


Biventricular support with HeartWare ventricular assist device in a pediatric patient.
Use of ventricular assist devices is increasing in the pediatric population. This has included the extended use of adult continuous-flow devices in the pediatric population. In a minority of cases, biventricular support may be needed. In these situations, biventricular support with continuous-flow devices can be surgically challenging, and therefore, only few cases have been reported. Here, we present a case of implantation of two HeartWare HVAD devices for biventricular support for a decompensating patient with acute myocarditis as well as present an alternative implantation surgical strategy.


Function of the cytochrome bc1-aa3 branch of the respiratory network in mycobacteria and network adaptation occurring in response to its disruption.
The aerobic electron transport chain in Mycobacterium smegmatis can terminate in one of three possible terminal oxidase complexes. The structure and function of the electron transport pathway leading from the menaquinol-menaquinone pool to the cytochrome bc1 complex and terminating in the aa3-type cytochrome c oxidase was characterized. M. smegmatis strains with mutations in the bc1 complex and in subunit II of cyctochome c oxidase were found to be profoundly growth impaired, confirming the importance of this respiratory pathway for mycobacterial growth under aerobic conditions. Disruption of this pathway resulted in an adaptation of the respiratory network that is characterized by a marked up-regulation of cydAB, which encodes the bioenergetically less efficient and microaerobically induced cytochrome bd-type menaquinol oxidase that is required for the growth of M. smegmatis under O2-limiting conditions. Further insights into the adaptation of this organism to rerouting of the electron flux through the branch terminating in the bd-type oxidase were revealed by expression profiling of the bc1-deficient mutant strain using a partial-genome microarray of M. smegmatis that is enriched in essential genes. Although the expression profile was indicative of an increase in the reduced state of the respiratory chain, blockage of the bc1-aa3 pathway did not induce the sentinel genes of M. smegmatis that are induced by oxygen starvation and are regulated by the DosR two-component regulator.


Monitoring DNA cytometric parameters during the course of chronic myelogenous leukemia.
The prognostic value of three DNA cytometric parameters--stemline ploidy (STL), stemline shoulder fraction (SSF) and "proliferative" fraction (PRF)--for the prediction of disease transformation and survival was examined for 20 patients with chronic myelogenous leukemia (CML) during the course of their disease and compared with two commonly used hematologic parameters (degree of leukocytosis and percentage of circulating leukemic progenitor cells). With disease progression, STL and SSF increased significantly, whereas PRF showed a steady decrease from diagnosis to blast crisis. The most significant part of these changes took place during the chronic phase, before the clinical onset of disease transformation. Hematologic parameters, in comparison, revealed significant changes later, shortly before blast crisis. The remaining duration of the chronic phase diminished from 25.5 months at the time of diagnosis, when the median STL was 2.0c, to 19.6 months for patients showing an STL of 2.1c, to 15.0 months with an STL of 2.2c and to 1.0 months for those with an STL of greater than or equal to 2.3c. Prognostically relevant limits for SSF and PRF were at 20%. When the SSF passed this limit or the PRF fell below it, the mean remaining chronic phase of these patients amounted to only 14.1 and 10.1 months. Interactive cytometry allows analysis of the DNA cytometric equivalent of changes in leukemic progenitor cells, which are well known from cytogenetic and cell kinetic studies. These three DNA cytometric parameters reflect the "natural history" of CML with the development of a cytogenetically hyperdiploid clone during disease progression in most patients and a simultaneous loss of proliferative potential on the level of myelobasts.(ABSTRACT TRUNCATED AT 250 WORDS)


Estimating the size of the HIV epidemic by using mortality data.
Evidence that more people are dying as a result of HIV infection than is reflected by the number of deaths among reported cases meeting the WHO definition of AIDS is derived from mortality data. Ninety-five causes of death likely to be associated with HIV infection were selected. Standardized mortality ratios due to these causes increased for single men aged 15-54 years from 100 in 1984 to 118 in 1987. The age, sex, marital status, temporal and geographic distribution of these excess deaths suggest that they are HIV-associated. It is estimated that 58% of excess deaths due to HIV-related causes were among cases reported to the CDSC AIDS Surveillance Programme in 1987. Some of these deaths may have been among HIV-positive people who did not meet the WHO definition at the time of death. There is a need for surveillance to be extended to include HIV-positive people who die before meeting the WHO definition if the full extent of the HIV epidemic is to be identified.


Jianpi Yangzheng Xiaozheng decoction alleviates gastric cancer progression via suppressing exosomal PD-L1.
Jianpi Yangzheng Xiaozheng decoction (JPYZXZ) is an empirical traditional Chinese medicine formula that has been reported to significantly prolong the survival of patients with advanced gastric cancer (GC). However, its underlying mechanism have not been fully elucidated. The present work aims to explore the possible mechanism of JPYZXZ on regulating GC progression. We firstly confirmed the inhibitory effect of JPYZXZ in GC MKN74 cells and 615-strain mice, which was possibly mediated with IL-6/JAK2/STAT3 pathway dependent PD-L1 expression. Moreover, we showed that JPYZXZ diminished the expression levels of GC-derived exosomal PD-L1 in MFC murine cells and xenograft GC model, as well as stage IIA-IIIB GC patients. We further found that in different types of tumor-infiltrating immune cells, PD-L1 expression was most positively correlated with myeloid-derived suppressor cells (MDSCs) in GC in the TISIDB database. We isolated exosomes derived from supernatants of MFC cells and co-cultured with bone marrow cells derived from C57BL/6 mice, and further revealed that the expansion of MDSCs was mediated by GC-derived exosomal PD-L1. Meanwhile, our results indicated that JPYZXZ inhibited the delivery of exosomal PD-L1 from GC cells to bone marrow cells, thereby alleviating exosomal PD-L1-induced differentiation and expansion of MDSCs in the tumor microenvironment. This led to a decrease in the levels of several immunosuppressive factors, including iNOS, Arg-1, TGF-β, IL-10, and IL-6, in 615-strain mice. Moreover, clinical data also revealed a significant positive relationship between exosomal PD-L1 and polymorphonuclear MDSCs under the JPYZXZ treatment in stage IIA-IIIB GC patients. In conclusion, our study confirmed that exosomal PD-L1 could be a key factor in controlling MDSCs differentiation in GC. JPYZXZ alleviated GC progression via suppressing exosomal PD-L1 mediated expansion of MDSCs, thereby remodeling the immunosuppressive tumor microenvironment, which provided the experimental evidence for the clinical application of JPYZXZ in the treatment of GC via PD-L1.