It is pleasant and heavy: convergence of visual contents in tobacco, alcohol and food marketing in Brazil.
The tactical use of visuoperceptual content in marketing may encourage impulsive consumption of unhealthy products. In this study, the application of visuoperceptual content was compared in advertisements used by industries of tobacco, alcohol and food. The aim was to ascertain whether similarities exist in the strategies used as variables for the selection of commercial stimuli, such as color, position and size. Scion Image and Corel Draw Graphics Suite software were used to analyze the content of a non-probabilistic sample of advertising images (N = 150). Differences were identified in the use of the colors green (p = 0.04) and red (p = 0.01), but not in the use of the color blue (p = 0.64), suggesting that induction of feelings of pleasantness resulting from the use of the color blue may be associated with the advertising in the alcohol and tobacco industries. Regarding the position of the commercial stimuli, a predominance of the use of quadrants 'C' (p = 0.00) and 'D' (p = 0.01) was found in all three industries, indicating a similar use of areas perceived as being 'heavier'. As to the size, 78% of advertisements placed the commercial stimuli within a range of 0-25% of the total image. The results showed some similarities in the use of visuoperceptual content in advertisements for tobacco, alcohol and food, especially between tobacco and alcohol. The article offers a convergence analysis of these three industries altogether, providing additional subsidies for the formulation of protection policies.


Controlled release from bilayer-decorated magnetoliposomes via electromagnetic heating.
Nanoscale assemblies that can be activated and controlled through external stimuli represent a next stage in multifunctional therapeutics. We report the formation, characterization, and release properties of bilayer-decorated magnetoliposomes (dMLs) that were prepared by embedding small hydrophobic SPIO nanoparticles at different lipid molecule to nanoparticle ratios within dipalmitoylphosphatidylcholine (DPPC) bilayers. The dML structure was examined by cryogenic transmission electron microscopy and differential scanning calorimetry, and release was examined by carboxyfluorescein leakage. Nanoparticle heating using alternating current electromagnetic fields (EMFs) operating at radio frequencies provided selective release of the encapsulated molecule at low nanoparticle concentrations and under physiologically acceptable EMF conditions. Without radio frequency heating, spontaneous leakage from the dMLs decreased with increasing nanoparticle loading, consistent with greater bilayer stability and a decrease in the effective dML surface area due to aggregation. With radio frequency heating, the initial rate and extent of leakage increased significantly as a function of nanoparticle loading and electromagnetic field strength. The mechanism of release is attributed to a combination of bilayer permeabilization and partial dML rupture.


Trigeminal neuropathic pain is alleviated by inhibition of Cav3.3 T-type calcium channels in mice.
In this brief report, we demonstrate that the Cav3.3 T-type voltage-gated calcium channel subtype is involved in our FRICT-ION model of chronic trigeminal neuropathic pain. We first showed that the Cacna1i gene encoding Cav3.3 is significantly upregulated in whole trigeminal ganglia of FRICT-ION mice compared to controls at week 10 post-injury. We confirmed protein upregulation of Cav3.3 compared to controls using Western blot analysis of whole trigeminal ganglia tissues. Finally, we demonstrated that intraperitoneal injection of a selective TAT-based Cav3.3 blocking peptide in FRICT-ION mice significantly reduces Cav3.3 protein expression at the peak anti-allodynic effect (4 hrs post-injection) of the attenuated neuropathic pain behavior. We also suggest that blockade of Cav3.3 may be more effective in attenuating trigeminal neuropathic pain in female than male FRICT-ION mice. Therefore, blocking or attenuating Cav3.3 function may be an effective strategy for the treatment of trigeminal neuropathic pain.


COVID-19 in Singapore and Malaysia: Rising to the Challenges of Orthopaedic Practice in an Evolving Pandemic.
With the increasing number of COVID-19 cases and related deaths worldwide, we decided to share the development of this condition in Singapore and Malaysia. First few cases were diagnosed in the two countries at the end of January 2020, and the numbers have surged to thousands by end of March 2020. We will focus on strategies adopted by the government and also the Orthopaedic community of the two countries up till the beginning of April 2020. We hope that by sharing of relevant information and knowledge on how we are managing the COVID-19 condition, we can help other communities, and health care workers to more effectively overcome this pandemic.


Evolution of phenolic compounds of spanish oak wood during natural seasoning. First results.
Low molecular weight polyphenols and ellagitannins were analyzed by HPLC, and the molecular weight distribution of ellagitannins was calculated by GPC, in oak heartwood of Quercus robur L., Quercus petraea Liebl., Quercus pyrenaica Wild., and Quercus faginea Lam., grown in Spain, before and after 1 year of seasoning, in Bordeaux, France. During this process, the concentrations of low molecular weight polyphenols (acids and aldehydes, benzoic and cinnamic, and coumarins) increased, and those of ellagitannins (castalagin, vescalagin, and roburins A-E) decreased. A similar behavior for the A and B compounds in all species was not found. This modification in the chemical composition was similar in the four Spanish species of Quercus studied and allowed the differentiation between the unseasoned wood and the wood after the first year of seasoning.


Effect of lower versus higher doses of tamoxifen on pituitary-gonadal function and sperm indices in oligozoospermic men.
Administration of the antiestrogen tamoxifen for one month to 12 patients with idiopathic oligozoospermia significantly increased the mean basal testosterone (T) level and the responses of luteinizing hormone (LH) and follicle stimulating hormone (FSH) to constant luteinizing hormone releasing hormone (LHRH) infusion but did not significantly influence the mean oestradiol (E2) levels or the E2 over testosterone ratio. Mean sperm concentration and total sperm output increased by about 70% after a mean treatment period of 5.5 +/- 0.4 months. No statistically significant difference was found between the two subgroups of patients treated with either the lower (5 or 10 mg once daily) or higher dose of tamoxifen (10 mg twice daily) with respect to basal or LHRH stimulated gonadotropin and testosterone response or the E2/T ratio and the effect on sperm density and total sperm output. In both subgroups the sperm motility and morphology remained unchanged. In conclusion higher doses of tamoxifen in this study prove not to be superior to lower doses in improving mean sperm density and total sperm output. The relative small percentage of patients achieving normalisation of only these sperm parameters pleads for further search for more effective selection of patients and other more effective treatment modalities in patients with idiopathic oligozoospermia.


Do working conditions explain the increased risks of disability pension among men and women with low education? A follow-up of Swedish cohorts.
Rates of disability pension are greatly increased among people with low education. This study examines the extent to which associations between education and disability pensions might be explained by differences in working conditions. Information on individuals at age 13 years was used to assess confounding of associations. Two nationally representative samples of men and women born in 1948 and 1953 in Sweden (22 889 participants in total) were linked to information from social insurance records on cause (musculoskeletal, psychiatric, and other) and date (from 1986-2008) of disability pension. Education data were obtained from administrative records. Occupation data were used for measurement of physical strain at work and job control. Data on paternal education, ambition to study, and intellectual performance were collected in school. Women were found to have higher rates of disability pension than men, regardless of diagnosis, whereas men had a steeper increase in disability pension by declining educational level. Adjustment of associations for paternal education, ambition to study, and intellectual performance at age 13 had a considerable attenuating effect, also when disability pension with a musculoskeletal diagnosis was the outcome. Despite this, high physical strain at work and low job control both contributed to explain the associations between low education and disability pensions in multivariable models. Working conditions seem to partly explain the increased rate of disability pension among men and women with lower education even though this association does reflect considerable selection effects based on factors already present in late childhood.


The cAMP Pathway Amplifies Early MyD88-Dependent and Type I Interferon-Independent LPS-Induced Interleukin-10 Expression in Mouse Macrophages.
Interleukin-10 (IL-10) is a key anti-inflammatory cytokine, secreted by macrophages and other immune cells to attenuate inflammation. Autocrine type I interferons (IFNs) largely mediate the delayed expression of IL-10 by LPS-stimulated macrophages. We have previously shown that IL-10 is synergistically expressed in macrophages following a costimulus of a TLR agonist and cAMP. We now show that the cAMP pathway directly upregulates IL-10 transcription and plays an important permissive and synergistic role in early, but not late, LPS-stimulated IL-10 mRNA and protein expression in mouse macrophages and in a mouse septic shock model. Our results suggest that the loss of synergism is not due to desensitization of the cAMP inducing signal, and it is not mediated by a positive crosstalk between the cAMP and type I IFN pathways. First, cAMP elevation in LPS-treated cells decreased the secretion of type I IFN. Second, autocrine/paracrine type I IFNs induce IL-10 promoter reporter activity only additively, but not synergistically, with the cAMP pathway. IL-10 promoter reporter activity was synergistically induced by cAMP elevation in macrophages stimulated by an agonist of either TLR4, TLR2/6, or TLR7, receptors which signal via MyD88, but not by an agonist of TLR3 which signals independently of MyD88. Moreover, MyD88 knockout largely reduced the synergistic IL-10 expression, indicating that MyD88 is required for the synergism displayed by LPS with cAMP. This report delineates the temporal regulation of early cAMP-accelerated vs. late type I IFN-dependent IL-10 transcription in LPS-stimulated murine macrophages that can limit inflammation at its onset.


A Rare Case of Monophasic Synovial Sarcoma of Thoracic Vertebra.
Synovial sarcoma of spine is an extremely rare malignancy with poor prognosis. It is often metastatic at the time of presentation. Its relative rarity and histological resemblance to other tumors make it diagnostically challenging, requiring the need of immunohistochemistry and cytogenetics for definite diagnosis. Surgery is the mainstay of therapy with adjunct chemotherapy, although survival rates are very low.


Oral oseltamivir treatment of influenza in children.
Oral oseltamivir administration is effective treatment for influenza in adults. This study was conducted to determine the efficacy, safety and tolerability of oseltamivir in children with influenza. In this randomized, double blind, placebo-controlled study, children 1 through 12 years with fever [> or =100 degrees F (> or =38 degrees C)] and a history of cough or coryza <48 h duration received oseltamivir 2 mg/kg/dose or placebo twice daily for 5 days. The primary efficacy endpoint was the time to resolution of illness including mild/absent cough and coryza mild/absent, return to normal activity and euthermia. Of 695 enrolled children 452 (65%) had influenza (placebo, n = 235; oseltamivir, n = 217). Among infected children the median duration of illness was reduced by 36 h (26%) in oseltamivir compared with placebo recipients (101 h; 95% confidence interval, 89 to 118 vs. 137 h; 95% confidence interval, 125 to 150; P < 0.0001). Oseltamivir treatment also reduced cough, coryza and duration of fever. New diagnoses of otitis media were reduced by 44% (12% vs. 21%). The incidence of physician-prescribed antibiotics was significantly lower in influenza-infected oseltamivir (68 of 217, 31%) than placebo (97 of 235, 41%; P = 0.03) recipients. Oseltamivir therapy was generally well-tolerated, although associated with an excess frequency of emesis (5.8%). Discontinuation because of adverse events was low in both groups (1.8% with oseltamivir vs. 1.1% with placebo). Oseltamivir treatment did not affect the influenza-specific antibody response. Oral oseltamivir administration is an efficacious and well-tolerated therapy for influenza in children when given within 48 h of onset of illness.


Prospects for quantitative and time-resolved double and continuous exposure off-axis electron holography.
The technique of double exposure electron holography, which is based on the superposition of two off-axis electron holograms, was originally introduced before the availability of digital image processing to allow differences between electron-optical phases encoded in two electron holograms to be visualised directly without the need for holographic reconstruction. Here, we review the original method and show how it can now be extended to permit quantitative studies of phase shifts that oscillate in time. We begin with a description of the theory of off-axis electron hologram formation for a time-dependent electron wave that results from the excitation of a specimen using an external stimulus with a square, sinusoidal, triangular or other temporal dependence. We refer to the more general method as continuous exposure electron holography, present preliminary experimental measurements and discuss how the technique can be used to image electrostatic potentials and magnetic fields during high frequency switching experiments.


Scholarly productivity of faculty in primary care roles related to tenure versus non-tenure tracks.
Increasing the number of primary care physicians is critical to overcoming the shortage of healthcare providers. Primary care physicians are increasingly called upon to address not only medical concerns but also behavioral health needs and social determinants of health which requires ongoing research and innovation. This paper evaluated scholarly productivity of faculty in tenure versus non-tenure tracks in primary care roles, defined as family medicine, internal medicine, internal medicine/pediatrics and pediatrics. The study included physician faculty in the clinical departments of Brody School of Medicine serving between the 2014-2015 and 2018-2019 academic years. Department, track, and rank at the beginning of each academic year (e.g., 2014-2015) were correlated with having any publications in the following calendar year (e.g., 2015), as determined from a search of the Scopus database. A total of 1620 observations and 542 unique faculty were included in the analysis. As of 2018-2019, 19% percent of primary care faculty were either tenured or on tenure track, as compared to 41% of faculty in other departments (p < 0.001). Primary care departments were also disproportionately staffed by junior faculty (60% as compared to 48% in other departments; p = 0.087). The proportion of faculty with any publications was significantly higher for faculty on the tenure track compared to those not on the tenure track (34% vs. 14%, p < 0.001). Academic productivity was lower among non-tenure-track physician faculty, as measured by publication in peer-reviewed journals. This was exacerbated among faculty in primary care departments, who were also more likely to hold non-tenure-track appointments. The loss of tenure-track positions disproportionately impacts scholarly activity in primary care and may be limiting progress in care-oriented research. Findings suggest that providing non-tenure faculty the time and resources to be involved in research, in addition to their clinical work, as well as access to research collaborators and mentors can promote scholarly activity among this group.


Propagation property of a nonuniformly polarized beam array in turbulent atmosphere.
A nonuniformly polarized beam array (NUBPA) is modeled by coherent superposition of a pair of orthogonally polarized spatial modes. The propagation of a NUPBA in turbulent atmosphere is investigated based on the extended Huygens-Fresnel method. An analytical expression for the intensity profile of a NUBPA in turbulent atmosphere is presented. The influence of polarization degree, intensity of turbulence, array number, and distance between adjacent elements on the intensity profile in the receiving plane is evaluated numerically and analyzed in detail.


Intracellular protein degradation in serum-deprived human fibroblasts.
IMR90 human fibroblasts were labelled by incubation of cells for 48 h in medium containing 10% serum and [3H]leucine. The labelled protein was degraded at a rate of 1%/h during a subsequent incubation in medium with 10% serum. Incubation in medium without serum caused a transient enhancement of the degradation of endogenous protein, which was also found in cells labelled in medium without serum. The degradation of micro-injected haemoglobin was enhanced by serum deprivation in a non-transient manner. These results suggest that enhanced degradation in serum-free medium occurs only for a subpopulation of cell proteins and that it appears transient because the major part of the pool of susceptible endogenous proteins is being degraded during the first 20-30 h in serum-free unlabelled medium. Protein turnover in various cell compartments was measured by a double-labelling technique. Most of the enhanced degradation in serum-deprived cultures (73-83%) was due to breakdown of cytosolic proteins. The enhanced degradation of cytosolic proteins seemed to affect several proteins irrespective of their molecular mass or metabolic stability.


Geldanamycin and herbimycin A induce apoptotic killing of B chronic lymphocytic leukemia cells and augment the cells' sensitivity to cytotoxic drugs.
We studied the actions of geldanamycin (GA) and herbimycin A (HMA), inhibitors of the chaperone proteins Hsp90 and GRP94, on B chronic lymphocytic leukemia (CLL) cells in vitro. Both drugs induced apoptosis of the majority of CLL isolates studied. Whereas exposure to 4-hour pulses of 30 to 100 nM GA killed normal B lymphocytes and CLL cells with similar dose responses, T lymphocytes from healthy donors as well as those present in the CLL isolates were relatively resistant. GA, but not HMA, showed a modest cytoprotective effect toward CD34+ hematopoietic progenitors from normal bone marrow. The ability of bone marrow progenitors to form hematopoietic colonies was unaffected by pulse exposures to GA. Both GA and HMA synergized with chlorambucil and fludarabine in killing a subset of CLL isolates. GA- and HMA-induced apoptosis was preceded by the up-regulation of the stress-responsive chaperones Hsp70 and BiP. Both ansamycins also resulted in down-regulation of Akt protein kinase, a modulator of cell survival. The relative resistance of T lymphocytes and of CD34+ bone marrow progenitors to GA coupled with its ability to induce apoptosis following brief exposures and to synergize with cytotoxic drugs warrant further investigation of ansamycins as potential therapeutic agents in CLL.


Health policy for the poor: an exploration on the take-up of means-tested health benefits in Turkey.
Recent healthcare reform in Turkey aims at achieving universal coverage with the introduction of General Health Insurance (GHI). As part of GHI, the state assumes the provision of health insurance coverage to those unable to afford the public health insurance premiums conditional on a means-testing procedure where the official threshold is set as one-third of the gross minimum wage. This article aims at exploring in Turkey the prevalence of non-take up of means-tested health insurance for the poor and the consequent financial burdens for those poor segments outside the coverage. Based upon Statistics of Income and Living Conditions micro data, the non-take-up rate is estimated to be around 44%, where the prevalence of non-take-up is lower yet still high, i.e. around 30%, for households with very low incomes as well as those with elderly or ill members. The results from a separate health expenditure survey on urban poor, which is specifically designed and implemented by the authors, reveal that poor households without health insurance coverage are faced with significant out-of-pocket expenditures. About 5% of those households without coverage were found to have inpatient expenditures that exceeded 20% of their annual disposable household income. Also, among the households without coverage but with at least one inpatient visit over the last two years, the median expenditure was reported as high as 8% of the annual household income as opposed to 0% median value for those with GHI. The results highlight that a large proportion of poor population still lacks public health insurance despite the overarching aim of universal coverage.


Effects of tryptophan depletion and a simulated alcohol binge on impulsivity.
Researchers have suggested binge drinkers experience disproportionate increases in impulsivity during the initial period of drinking, leading to a loss of control over further drinking, and that serotonergic mechanisms may underlie such effects. We examined the effects of a simulated alcohol binge and tryptophan depletion on 3 types of impulsivity-response initiation (immediate memory task [IMT]), response inhibition (GoStop task), and delay discounting (single key impulsivity paradigm [SKIP])-and tested whether observed effects were related to real-world binging. Adults (N = 179) with diverse drinking histories completed a within-subject crossover design over 4 experimental days. Each day, participants underwent 1 of 4 test conditions: tryptophan depletion/alcohol, tryptophan depletion/placebo, tryptophan-balanced control/alcohol, or tryptophan-balanced control/placebo. The simulated binge involved consuming 0.3 g/kg of alcohol at 5, 6, and 7 hr after consuming the tryptophan-depletion/balanced mixture. Impulsivity was measured before and after each drink. Relative to the placebo beverage condition, when alcohol was consumed, impulsive responding was increased at moderate and high levels of intoxication on the IMT and the GoStop but only at high levels of intoxication on the SKIP. Tryptophan depletion had no effect on impulsivity. Effects of alcohol and tryptophan manipulations on impulsivity were unrelated to patterns of binge drinking outside the laboratory. The effects of alcohol consumption on impulsivity depend on the component of impulsivity and the dose of alcohol consumed. Such effects do not appear to be a result of reduced serotonin synthesis. In addition, real-world binge drinking behaviors were unrelated to behavioral changes observed in the laboratory.


[Toxicity and tolerance of toxogonin].
A comparative characteristic of acute toxicity of toxogonine and dipiroxime is given for different animal species with various routes of introduction of the compounds from which it follows that, as concerns the toxicity parameters and the area of toxic action, toxogonine is in the lead. It is also shown that with its multiple administration in a therapeutic dose toxogonine does not affect the peripheral blood, the hepatic and renal functions, the cardio-vascular system, this being also supported by the anatomo-pathological findings. With its intramuscular introduction toxogonine does not produce any local irritation.


An intelligent cell-selective polymersome-DM1 nanotoxin toward triple negative breast cancer.
Antibody-drug conjugates (ADCs) are among the most significant advances in clinical cancer treatments, however, they are haunted with fundamental issues like low drug/antibody ratio (DAR), need of large amount of antibody, and complex chemistry. Targeted nanomedicines while offering a promising alternative to ADCs are afflicted with drug leakage and inferior cancer-specificity. Herein, we developed an intelligent cell-selective nanotoxin based on anti-CD44 antibody-polymersome-DM1 conjugates (aCD44-AP-DM1) for potent treatment of solid tumors. DM1 was simultaneously coupled to vesicular membrane via disulfide bonds during self-assembly and anti-CD44 antibody was facilely clicked onto polymersome surface, tailor-making an optimal aCD44-AP-DM1 with a controlled antibody density of 5.0, extraordinary DAR of 275, zero drug leakage and rapid reduction-responsive DM1 release. aCD44-AP-DM1 displayed a high specificity and exceptional cytotoxicity toward MDA-MB-231 triple negative breast cancer, SMMC-7721 hepatocellular carcinoma and A549 non-small cell lung cancer cells with half-maximal inhibitory concentrations (IC50) of 21.4, 3.7 and 64.6 ng/mL, respectively, 3.6-47.2-fold exceeding non-targeted P-DM1. Intriguingly, the systemic administration of aCD44-AP-DM1 significantly suppressed subcutaneous MDA-MB-231 tumor xenografts in nude mice while intratumoral injection achieved complete tumor eradication in four out of five mice, without causing toxicity. This intelligent cell-selective nanotoxin has emerged as a better platform over ADCs for targeted cancer therapy.


Commentary. Major drug-prescribing patterns in general hospitals.
Prescriptions of major drug classes were surveyed for one year in five teaching and five nonteaching general hospitals. Their overall rates varied over a two- to three-fold range in medicine, surgery, and pediatrics, but were uniform throughout the hospitals. Parenteral fluids and antibiotics represented over one half of the pharmacotherapy used in every specialty except medicine, where tranquilizers were the most frequently prescribed drugs. Prescribing patterns lacked consistency in obstetrics and newborn services, and generally with steroids, vasopressors, and anticoagulants. Significant variations also involved the use of fluids in pediatrics, diuretics in surgery, and vasodilators in medicine. These variations were too unpredictable and large to be attributed to specific differences among patient populations. Only two discrepancies were noted between teaching and nonteaching hospitals: tranquilizers were prescribed more often in the former, and vasopressors in the latter.


Colorectal Cancer in Inflammatory Bowel Disease.
Patients with inflammatory bowel disease (IBD) are at an increased risk for developing colorectal cancer (CRC). However, the incidence has declined over the past 30 years, which is probably attributed to raise awareness, successful CRC surveillance programs and improved control of mucosal inflammation through chemoprevention. The risk factors for IBD-related CRC include more severe disease (as reflected by the extent of disease and the duration of poorly controlled disease), family history of CRC, pseudo polyps, primary sclerosing cholangitis, and male sex. The molecular pathogenesis of inflammatory epithelium might play a critical role in the development of CRC. IBD-related CRC is characterized by fewer rectal tumors, more synchronous and poorly differentiated tumors compared with sporadic cancers. There is no significant difference in sex distribution, stage at presentation, or survival. Surveillance is vital for the detection and subsequently management of dysplasia. Most guidelines recommend initiation of surveillance colonoscopy at 8 to 10 years after IBD diagnosis, followed by subsequent surveillance of 1 to 2 yearly intervals. Traditionally, surveillance colonoscopies with random colonic biopsies were used. However, recent data suggest that high definition and chromoendoscopy are better methods of surveillance by improving sensitivity to previously "invisible" flat dysplastic lesions. Management of dysplasia, timing of surveillance, chemoprevention, and the surgical approaches are all areas that stimulate various discussions. The aim of this review is to provide an up-to-date focus on CRC in IBD, from laboratory to bedside.


Enhancing the Therapeutic Delivery of Oligonucleotides by Chemical Modification and Nanoparticle Encapsulation.
Oligonucleotide (ON) drugs, including small interfering RNA (siRNA), microRNA (miRNA) and antisense oligonucleotides, are promising therapeutic agents. However, their low membrane permeability and sensitivity to nucleases present challenges to in vivo delivery. Chemical modifications of the ON offer a potential solution to improve the stability and efficacy of ON drugs. Combined with nanoparticle encapsulation, delivery at the site of action and gene silencing activity of chemically modified ON drugs can be further enhanced. In the present review, several types of ON drugs, selection of chemical modification, and nanoparticle-based delivery systems to deliver these ON drugs are discussed.


Targeting mantle cell lymphoma metabolism and survival through simultaneous blockade of mTOR and nuclear transporter exportin-1.
Mantle cell lymphoma (MCL) is an aggressive B-cell lymphoma with poor prognosis, characterized by aberrant expression of growth-regulating and oncogenic effectors and requiring novel anticancer strategies. The nuclear transporter exportin-1 (XPO1) is highly expressed in MCL and is associated with its pathogenesis. mTOR signaling, a central regulator of cell metabolism, is frequently activated in MCL and is also an important therapeutic target in this cancer. This study investigated the antitumor effects and molecular/metabolic changes induced by the combination of the small-molecule selective inhibitor XPO1 inhibitor KPT-185 and the dual mTORC1/2 kinase inhibitor AZD-2014 on MCL cells. AZD-2014 enhanced the KPT-185-induced inhibition of cell growth and repression of cell viability. The combination of KPT-185 and AZD-2014 downregulated c-Myc and heat shock factor 1 (HSF1) with its target heat shock protein 70 (HSP70). As a consequence, the combination caused repression of ribosomal biogenesis demonstrated by iTRAQ proteomic analyses. Metabolite assay by CETOF-MS showed that AZD-2014 enhanced the KPT-185-induced repression of MCL cellular energy metabolism through the TCA (Krebs) cycle, and further repressed KPT-185-caused upregulation of glycolysis.Thus the simultaneous inhibition of XPO1 and mTOR signaling is a novel and promising strategy targeting prosurvival metabolism in MCL.


A case of adult-onset tubulointerstitial nephritis and uveitis syndrome presenting with granulomatous panuveitis.
The tubulointerstitial nephritis and uveitis (TINU) syndrome is a rare disorder that is probably underdiagnosed in clinical practice. Ocular involvement in TINU syndrome not only presents with the nongranulomatous anterior uveitis in 80% of patients but also manifests as intermediate, posterior, or panuveitis. This case report mentions an adult male patient who presented with granulomatous iridocyclitis with panuveitis and mild renal insufficiency. Workup for connective tissue and infectious diseases was negative for the patient. He was diagnosed with TINU syndrome based on the findings of renal biopsy. Both the uveitis and nephritis promptly responded well to steroid treatment, and there was no recurrence during the follow-up of 24 months.


Factors Influencing Sexual Function in Postmenopausal Married Women.
This study identified the level of marital intimacy, menopausal symptom and sexual function, and examined factors affecting sexual function in postmenopausal married women. With cross-sectional survey, a sample of 245 subjects aged 45 to 60 years was recruited from January to March, 2014. Data were collected using self-reported questionnaires which included marital intimacy questionnaire (MIQ), menopause rating scale (MRS), and female sexual function index (FSFI). Data were analyzed by descriptive statistics, t-test, ANOVA, Pearson correlation coefficients and stepwise multiple regression. Marital intimacy, menopausal symptom and sexual function scores of subjects were 3.46±1.14, 13.71±6.61, and 12.55±8.33, respectively. Marital intimacy and menopausal symptom were related to sexual function. Variables including marital intimacy, psychosocial and urogenital menopausal symptom, period since menopause and body mass index therapy explained 51.5% in sexual function of participants. When nurses plan sexual counseling and education for sexual function of postmenopausal married women, it is necessary to consider marital intimacy, menopausal symptoms, and menopause-related factors.


Social capital, trust in health information, and acceptance of Measles-Rubella vaccination campaign in Tamil Nadu: A case-control study.
Parents' decision about vaccination of children is influenced by social relationships and sources of information. The aim of this study was to assess the influence of social capital and trust in health information on the status of Measles-Rubella (MR) vaccination campaign in Tamil Nadu. This was a case-control study carried out in Kancheepuram district in Tamil Nadu where the MR vaccination campaign offered by Government of Tamil Nadu had poor acceptance. Cases were parents of children who had refused the MR vaccine and controls were parents having children in the same age group who had accepted the vaccine. Data on social capital and trust in health information were collected by using social capital scale developed by the researchers and trust in the source of information was measured by using simple questions on the level of trust in the information source. Nonadministration of MR vaccine was high among young parents and parents of younger children. Vaccine acceptance was higher when it was offered at school (P < 0.000) and also among parents who trusted school teachers (P < 0.003) and other school children (P < 0.014) as source of information. MR vaccine acceptance was less among parents who trusted social media and WhatsApp information. Greater levels of health-related physical social capital led to greater vaccine hesitancy. Multivariate analysis revealed that greater the age of the child, better parental attitudes toward vaccination, poorer health-related physical social capital, and greater trust in health information provided by school teachers led to overall greater acceptance of the MR vaccine. Strong homogeneous bonding social capital had a negative influence on MR vaccine acceptance. Schools and school teachers played a vital role in influencing parental decision to vaccinate.


A systematic review of the relationship between high school dropout and substance use.
A systematic review of peer-reviewed, empirical literature published between 1990 and 2006 was undertaken to determine whether existing research could provide evidence, and a deeper understanding of the relationship between dropping out of high school and the use of substances such as tobacco, alcohol, cannabis/marijuana and other illicit drugs. Forty-six articles were reviewed. The review describes the heterogeneity of theoretical frameworks employed, as well as the limited ability of any one to adequately explain the relationship between high school dropout and substance use. A refinement of the many confounding and mediating variables into coherent conceptual categories would aid more robust theory building and theory integration. In spite of differences in dropout definitions and diverse measures of substance use across studies, the main findings point to a largely consistent relationship between dropping out of high school and substance use. However, socially disadvantaged and poor persons, dropouts, and drug users are over-represented in some of the loss to follow-up groups in longitudinal studies surveyed. More rigorous mechanisms to retain participants in longitudinal studies should be employed. Suggestions for future research include comparisons between urban and rural populations, employing qualitative research methods, and research in developing countries, which have the least favourable school outcomes and a dearth of research on high school dropout.


Herbal Medicines: Malaysian Women's Knowledge and Practice.
This is a cross-sectional, descriptive study among Malay women admitted in the antenatal and postnatal ward to determine the prevalence and use of herbal medicines during pregnancy and elemental analysis in the most popular herbs. A total of 460 women were surveyed. Herbal medicine use during pregnancy was 34.3%, while 73% utilized herbal medicines during labor, because of a belief that it may shorten and ease labor. The most commonly used herbal medicines in pregnancy were Anastatica hierochuntica L. (60.1%) followed by coconut oil (35.4%). The majority of women (89.2%) used only one type of herbal medicines and took one capsule/glass (38%) per day. Herbal medicines use by pregnant women is largely unsupervised (81%), with most women getting information from their parents (60.7%) and buying the products directly from traditional midwives (32.2%) and 77% agreed upon its efficacy and safety. From the 460 respondents, 89.8% women were in the low end of the herbs knowledge. There was a significant difference found between knowledge score and income (P < 0.05). Microdiffraction analysis revealed significant presence of carbon, oxygen, silica, calcium, magnesium, aluminium, potassium, zinc, and iron that were found in Anastatica hierochuntica L. and proved to have good benefits for pregnancy.


Automated detection of physiologic deterioration in hospitalized patients.
Develop and evaluate an automated case detection and response triggering system to monitor patients every 5 min and identify early signs of physiologic deterioration. A 2-year prospective, observational study at a large level 1 trauma center. All patients admitted to a 33-bed medical and oncology floor (A) and a 33-bed non-intensive care unit (ICU) surgical trauma floor (B) were monitored. During the intervention year, pager alerts of early physiologic deterioration were automatically sent to charge nurses along with access to a graphical point-of-care web page to facilitate patient evaluation. Nurses reported the positive predictive value of alerts was 91-100% depending on erroneous data presence. Unit A patients were significantly older and had significantly more comorbidities than unit B patients. During the intervention year, unit A patients had a significant increase in length of stay, more transfers to ICU (p = 0.23), and significantly more medical emergency team (MET) calls (p = 0.0008), and significantly fewer died (p = 0.044) compared to the pre-intervention year. No significant differences were found on unit B. We monitored patients every 5 min and provided automated pages of early physiologic deterioration. This before-after study found a significant increase in MET calls and a significant decrease in mortality only in the unit with older patients with multiple comorbidities, and thus further study is warranted to detect potential confounding. Moreover, nurses reported the graphical alerts provided information needed to quickly evaluate patients, and they felt more confident about their assessment and more comfortable requesting help.


The effect of reimbursement policies on the management of Medicare patients with refractory ovarian cancer.
Ovarian cancer patients who are covered by Medicare are faced with therapeutic decisions that require consideration of out-of-pocket costs for oral anticancer agents and complete reimbursement for more expensive intravenous and often more toxic medications. The response rates for oral agents such as altretamine or etoposide are similar to those for intravenous paclitaxel or topotecan (14% to 26%), but the economic considerations differ markedly. Under current legislation, Medicare will completely cover the costs for the two intravenous outpatient chemotherapy regimens, but does not provide any financial support for oral regimens that do not have associated injectable formulations. This is a matter of concern for patients, as 89% prefer oral therapies. We compared the out-of-pocket costs and costs to the Medicare system of oral and intravenous agents used for refractory ovarian cancer, using published phase II and phase III data. The total cost of treatment was $18,635 for topotecan, $15,767 for paclitaxel, $7,721 for etoposide, and $4,477 for altretamine. Conversely, out-of-pocket costs for Medicare patients without Medigap coverage were highest for altretamine, at the full cost of $4,477, whereas Medicare covered all but $83 for topotecan, $37 for paclitaxel, and $66 for etoposide. Current Medicare reimbursement policies may affect patient options for cancer care. These policies are changing and should continue to change as Medicare adopts more managed care strategies.


Defining the Design Parameters for in Vivo Enzyme Delivery Through Protein Spherical Nucleic Acids.
The translation of proteins as effective intracellular drug candidates is limited by the challenge of cellular entry and their vulnerability to degradation. To advance their therapeutic potential, cell-impermeable proteins can be readily transformed into protein spherical nucleic acids (ProSNAs) by densely functionalizing their surfaces with DNA, yielding structures that are efficiently taken up by cells. Because small structural changes in the chemical makeup of a conjugated ligand can affect the bioactivity of the associated protein, structure-activity relationships of the linker bridging the DNA and the protein surface and the DNA sequence itself are investigated on the ProSNA system. In terms of attachment chemistry, DNA-based linkers promote a sevenfold increase in cellular uptake while maintaining enzymatic activity in vitro as opposed to hexaethylene glycol (HEG, Spacer18) linkers. Additionally, the employment of G-quadruplex-forming sequences increases cellular uptake in vitro up to fourfold. When translating to murine models, the ProSNA with a DNA-only shell exhibits increased blood circulation times and higher accumulation in major organs, including lung, kidney, and spleen, regardless of sequence. Importantly, ProSNAs with an all-oligonucleotide shell retain their enzymatic activity in tissue, whereas the native protein loses all function. Taken together, these results highlight the value of structural design in guiding ProSNA biological fate and activity and represent a significant step forward in the development of intracellular protein-based therapeutics.


Thyroglobulin messenger ribonucleic acid levels in the peripheral blood of children with benign and malignant thyroid disease.
Reverse transcriptase-PCR has identified thyroglobulin mRNA (Tg mRNA) in peripheral blood of normal adults and adults with thyroid cancer. However, no children were studied. The primary objective of this study was to determine whether whole blood Tg mRNA levels differ between benign and malignant thyroid disease in children. The secondary goals were to determine whether whole blood Tg mRNA levels vary with age or pubertal development among children with thyroid disease. Whole blood Tg mRNA levels were determined in 38 children (29 girls, nine boys; median age, 14.5 y; range, 4.8-20.4 y) with benign and malignant thyroid disease and correlated with diagnosis, age, pubertal status, thyroid size, and serum levels of free thyroxine, TSH, and Tg protein. Tg mRNA levels ranged from 3.3 to 104 pg Eq/microg total thyroid RNA (mean, 28 +/- 20.2 pg Eq/microg total thyroid RNA) and were similar in benign and malignant disorders (p = 0.67). However, in children with previously treated papillary thyroid cancer, Tg mRNA levels directly correlated with total body (131)I uptake (p = 0.026) and serum Tg protein (p = 0.037). There was no difference between boys and girls, and no change with pubertal maturation. In children with benign thyroid disease, Tg mRNA levels correlated with serum TSH (p = 0.031), but not with diagnosis, age, Tanner stage, or thyroid size. We conclude that Tg mRNA levels are similar in children with benign and malignant thyroid disease and unchanged by age or pubertal status, but correlated with tumor burden in previously treated papillary thyroid cancer.


MiR-526b-3p enhances sensitivity of head and neck squamous cell carcinoma cells to radiotherapy via suppressing exosomal LAMP3-mediated autophagy.
Lysosomal associated membrane protein 3 (LAMP3) has been reported to be a tumour promoter in multiple cancer types by modulating tumour cell autophagy. However, the potential mechanism of LAMP3 in radio-resistance of head and neck squamous cell carcinoma (HNSCC) remains unknown. Therefore, our current study aims to detect the impacts of LAMP3 on the resistance of HNSCC cells to radiotherapy and meanwhile explore its functional mechanism. Through RT-Qpcr examination, LAMP3 expression was identified to be expressed at a significantly high level in irradiation-resistant HNSCC cell lines compared with irradiation-sensitive HNSCC cell lines. Functional assays including CCK-8, colony formation and Transwell assays demonstrated that LAMP3 enhanced the radio-resistance through inducing autophagy to promote HNSCC cell growth. Furthermore, irradiation-resistant HNSCC cells could transfer exosomal LAMP3 to elevate LAMP3 expression in irradiation-sensitive HNSCC cells. Mechanistically, microRNA (miRNA) miR-526b-3p could inhibit LAMP3 expression so as to strengthen sensitivity of HNSCC cells to radiotherapy. In a word, exosomal LAMP3 expression promoted radioresistance of HNSCC cells via inducing autophagy, while this effect could be suppressed by miR-526b-3p in a targeted manner.


Stereoisomerization of beta-hydroxy-alpha-sulfenyl-gamma-butyrolactones controlled by two concomitant 1,4-type nonbonded sulfur-oxygen interactions as analyzed by X-ray crystallography.
We have synthesized nine beta-hydroxy-alpha-sulfenyl-gamma-butyrolactones having different substituents. The syn-anti or syn-syn lactones (or any mixture of both) invariably isomerized into syn-anti/syn-syn lactones in a ratio of approximately 6/4. The other two possible isomeric lactones (anti-syn or anti-anti) were never observed. The crystal structures of syn-syn lactones have been determined. We found sulfur-oxygen distances to be less than the sum of the van der Waals radii (3.3 A), with the angle formed among the hydroxylic oxygen, sulfur, and quaternary aromatic carbon being close to 180 degrees. In addition, carbonylic oxygen-sulfur is directed <40 degrees from the perpendicular to the C-S-C. Theoretical calculations were performed which emphasize the directionality of the sulfur-oxygen interaction. The X-ray and theoretical studies demonstrate that two concomitant, attractive 1,4 intramolecular interactions of divalent sulfur with both carbonyl and hydroxyl oxygens are the driving force for the aforementioned stereochemical preference. Then nonbonded sulfur-oxygen interactions would control the stereoselectivity of the reaction. The same features are observed in the X-ray structure of a beta-hydroxy-alpha-sulfinyl-gamma-butyrolactone.


Development of a microtitre plate enzyme immunoassay for the determination of progesterone.
A rapid, solid-phase microtitre plate enzyme immunoassay (EIA) for progesterone is described using progesterone 3-O-carboxymethyloxime-horseradish peroxidase as the label and an antiserum raised in rabbits to a progesterone 11 alpha-hemisuccinyl-bovine serum albumin immunogen. A competitive reaction was used with a reaction time of 2 h. Antibody-bound and free steroid were separated in a simple washing step of the antibody-adsorbed well surface. 2,2'-Azino-di-(3-ethylbenzthiazoline sulphonic acid) diammonium salt was used as the substrate with a reaction time of 1 h. A lower limit of sensitivity of 0.25 pg/well was obtained with the response being linear (logit/log) through 1000 pg/well. Results obtained by EIA and radioimmunoassay in several species gave excellent agreement (r = 0.98). This assay system allows accurate determination of progesterone in plasma with good specificity, precision and accuracy, and is suitable for the rapid assessment of luteal function and reproductive status in both clinical and research situations in a wide variety of species.


Parathyroid carcinoma masquerading as morning sickness in pregnancy.
Incidence of primary hyperparathyroidism (PHP) in pregnancy is 8/100,000 population/year with less than 200 cases reported. Physiological changes associated with pregnancy make a diagnosis of PHP difficult and 80% are asymptomatic. High index of suspicion is required as physiological hypocalcemia related to hemodilution, increased glomerular filtration rate resulting in maternal hypercalciuria and gestational hypoalbuminemia can mask hypercalcemia of PHP. Maternal and fetal complication rates are high. Early recognition followed by appropriate management and treatment significantly reduces complications. Here, we present a rare case of parathyroid carcinoma in pregnancy and highlight the difficulties in diagnosis given the non-specific symptoms related to hypercalcemia. We have also discussed the management of PHP during the pregnancy. PHP is a preventable cause of fetal and maternal morbidity and mortality.


Is mode of delivery associated with postpartum depression at 6 weeks: a prospective cohort study.
To examine the relationship between delivery mode and postpartum depression at 6 weeks following hospital discharge. A prospective cohort study. Eleven hospitals in Ontario, Canada. A total of 2560 women ≥16 years of age who delivered singleton, live infants at term. Women completed a questionnaire in hospital and 74% (n = 1897) participated in a structured telephone interview 6 weeks after discharge. Additional data were extracted from labour and delivery records. Generalised estimating equations (GEEs) were used to investigate factors associated with postpartum depression. Women were screened for depression at 6 weeks following hospital discharge using the Edinburgh Postnatal Depression Scale (EPDS). A score of ≥12 on the EPDS was used as a measure of the primary outcome, depression. Mode of delivery was not independently associated with postpartum depression, and did not factor into the main-effects model. The multivariable analysis identified 11 predictor variables for depression: young maternal age (OR 5.27; 95% CI 2.73-10.15); maternal hospital readmission (OR 3.02; 95% CI 1.46-6.24); non-initiation of breastfeeding (OR 2.02; 95% CI 0.99-4.11); good, fair, or poor self-reported postpartum health (OR 1.82; 95% CI 1.19-2.80); urinary incontinence (OR 1.79; 95% CI 1.06-3.03); multiparity (OR 1.59; 95% CI 1.22-2.08); low mental health functioning (OR 1.20; 95% CI 1.15-1.25); low subjective social status (OR 1.16; 95% CI 1.02-1.33); high number of unmet learning needs in hospital (OR 1.12; 95% CI 1.03-1.22); low social support (OR 1.06; 95% CI 1.03-1.09); and low physical health functioning (OR 1.03; 95% CI 1.003-1.055). An exploratory interaction model revealed that caesarean section was associated with higher odds of becoming depressed in Canadian-born women, but that in women born outside of Canada it was associated with a lower risk of becoming depressed. Delivery mode had no significant impact on the development of postpartum depression in the main-effects model. However, it may interact with place of birth and other unmeasured factors to create a risk for depression.


Characterization of the c-type cytochromes of Nitrosomonas europaea with the aid of fluorescent gels.
When a total soluble extract of Nitrosomonas europaea was denatured with dodecyl sulphate, subjected to dodecyl sulphate/polyacrylamide-gel electrophoresis and illuminated with near-u.v. light, eight bands of protein fluorescence were observed. All but one of these bands were red in colour, a property characteristic of c-type cytochromes. Standard techniques were used to purify soluble c-type cytochromes from this organism, and it was then possible to assign all but two very minor bands to specific c-type cytochromes, namely hydroxylamine oxidase, cytochrome c-554, cytochrome c-552 and a cytochrome c-550 not previously described. The eight band had fluorescence peaking in the green region of the spectrum, probably caused by covalently bound flavin, and co-purified with hydroxylamine oxidase. The following physical properties were determined for these components: isoelectric point, molecular weights according to gel filtration and mobility on dodecyl sulphate/polyacrylamide gels, and alpha-band spectra at room temperature and 77K. Redox potentials were measured as follows: cytochrome c-554, E(m,7) = +20mV; cytochrome c-552, E(m,7) = +230mV; cytochrome c-550, E(m,7) = +140mV. When washed membranes were applied to dodecyl sulphate/polyacrylamide gels in the same way, a number of fluorescent bands were observed that could be matched by soluble proteins. In addition, there was one band that could not be detected in supernatants, migrating with an apparent molecular weight of 24000. This species is probably coincident with a c-type cytochrome having E(m,7) = +170mV found in redox titration of these membranes. In future studies, gel fluorescence should form a useful complement to spectroscopy for analysis of cytochrome composition in active cell-free preparations or semi-purified material.


Alpha subunit in gestational trophoblastic disease.
Alpha HCG, estimated after evacuation of hydatidiform mole, was found to follow the decline of beta-HCG. In patients with low-risk non-metastatic gestational disease the alpha-subunit values also followed the beta-HCG values. This indicates no additional value in following alpha-HCG in such patients. Whether alpha-HCG elevation may portend recurrence in treated high-risk cases of metastatic disease in remission with non-detectable beta-HCG remains unresolved by this study.


Molecular dynamics simulations of polymer welding: strength from interfacial entanglements.
Large-scale simulations of thermal welding of polymers are performed to investigate the rise of mechanical strength at the polymer-polymer interface with the welding time t(w). The welding process is at the core of integrating polymeric elements into devices as well as in the thermal induced healing of polymers, processes that require the development of interfacial strength equal to that of the bulk. Our simulations show that the interfacial strength saturates at the bulk shear strength long before polymers diffuse by their radius of gyration. Along with the strength increase, the dominant failure mode changes from chain pullout at the interface to chain scission as in the bulk. The formation of sufficient entanglements across the interface, which we track using a primitive path analysis, is required to arrest catastrophic chain pullout at the interface. The bulk response is not fully recovered until the density of entanglements at the interface reaches the bulk value. Moreover, the increase of interfacial strength before saturation is proportional to the number of interfacial entanglements between chains from opposite sides.


Cell-mass structures expressing the aromatase gene Cyp19a1 lead to ovarian cavities in Xenopus laevis.
The African clawed frog, Xenopus laevis, has a ZZ/ZW-type sex-determination system. We previously reported that a W-linked gene, Dm-W, can determine development as a female. However, the mechanisms of early sex differentiation remain unclear. We used microarrays to screen for genes with sexually dimorphic expression in ZZ and ZW gonads during early sex differentiation in X laevis and found several steroidogenic genes. Importantly, the steroid 17α-hydroxylase gene Cyp17a1 and the aromatase gene Cyp19a1 were highly expressed in ZZ and ZW gonads, respectively, just after sex determination. At this stage, we found that Cyp17a1, Cyp19a1, or both were expressed in the ZZ and ZW gonads in a unique mass-in-line structure, in which several masses of cells, each surrounded by a basement membrane, were aligned along the anteroposterior axis. In fact, during sex differentiation, ovarian cavities formed inside each mass of Cyp17a1- and Cyp19a1-positive cells in the ZW gonads. However, the mass-in-line structure disappeared during testicular development in the ZZ testes. These results suggested that the mass-in-line structure found in both ZZ and ZW gonads just after sex determination might be formed in advance to produce ovarian cavities and then oocytes. Consequently, we propose a view that the default sex may be female in the morphological aspect of gonads in X laevis.


Beyond scale-free networks: integrating multilayer social networks with molecular clusters in the local spread of COVID-19.
This study evaluates the scale-free network assumption commonly used in COVID-19 epidemiology, using empirical social network data from SARS-CoV-2 Delta variant molecular local clusters in Houston, Texas. We constructed genome-informed social networks from contact and co-residence data, tested them for scale-free power-law distributions that imply highly connected hubs, and compared them to alternative models (exponential, log-normal, power-law with exponential cutoff, and Weibull) that suggest more evenly distributed network connections. Although the power-law model failed the goodness of fit test, after incorporating social network ties, the power-law model was at least as good as, if not better than, the alternatives, implying the presence of both hub and non-hub mechanisms in local SARS-CoV-2 transmission. These findings enhance our understanding of the complex social interactions that drive SARS-CoV-2 transmission, thereby informing more effective public health interventions.


Differentiation-independent fluctuation of pluripotency-related transcription factors and other epigenetic markers in embryonic stem cell colonies.
Embryonic stem cells (ESCs) maintain their pluripotency through high expression of pluripotency-related genes. Here, we show that differing levels of Oct4, Nanog, and c-myc proteins among the individual cells of mouse ESC (mESC) colonies and fluctuations in these levels do not disturb mESC pluripotency. Cells with strong expression of Oct4 had low levels of Nanog and c-myc proteins and vice versa. In addition, cells with high levels of Nanog tended to occupy interior regions of mESC colonies. In contrast, peripherally positioned cells within colonies had dense H3K27-trimethylation, especially at the nuclear periphery. We also observed distinct levels of endogenous and exogenous Oct4 in particular cell cycle phases. The highest levels of Oct4 occurred in G2 phase, which correlated with the pKi-67 nuclear pattern. Moreover, the Oct4 protein resided on mitotic chromosomes. We suggest that there must be an endogenous mechanism that prevents the induction of spontaneous differentiation, despite fluctuations in protein levels within an mESC colony. Based on the results presented here, it is likely that cells within a colony support each other in the maintenance of pluripotency.


[Kangaroo-Mother Care: scientific evidence and impact on breastfeeding].
To describe the history of the Kangaroo Mother Care and present scientific evidence about benefits of this practice on morbidity and mortality, psychological and neurological development and breastfeeding of low birth weight infants. Papers about Kangaroo Mother Care published from 1983 to 2004 were consulted, selected in Medline and Lilacs, as well as books, thesis and technical publications from the Brazilian Health Department. Since its first description, Kangaroo Mother Care has been extensively studied. The analysis of randomized trials showed that it consists in a protection factor to breastfeeding at discharge (relative risk 0.41, 95% confidence interval 0.25 to 0.68). The method was always associated with the following reduced risks: nosocomial infection at 41 weeks' corrected gestational age (relative risk 0.49, 95% confidence interval 0.25 to 0.93), severe illness (relative risk 0.30, 95% confidence interval 0.14 to 0.67), lower respiratory tract disease at 6 months (relative risk 0.37, 95% confidence interval 0.15 to 0.89) and better gain of weight per day (weighted mean difference 3.6 g/day, 95% confidence interval 0.8 to 6.4). Psychomotor development at 12 months' corrected age was similar in the two groups. There was no evidence of a difference in infant's mortality. A positive impact of Kangaroo Mother Care on breastfeeding was found. Although the method appears to reduce severe infant morbidity without any serious deleterious effect reported, there is still insufficient evidence to recommend its routine use. It is necessary to develop studies about effectiveness, acceptability and applicability of the method in the Brazilian context.


Fc-based Duokines: dual-acting costimulatory molecules comprising TNFSF ligands in the single-chain format fused to a heterodimerizing Fc (scDk-Fc).
Targeting costimulatory receptors of the tumor necrosis factor superfamily (TNFSF) to activate T-cells and promote anti-tumor T-cell function have emerged as a promising strategy in cancer immunotherapy. Previous studies have shown that combining two different members of the TNFSF resulted in dual-acting costimulatory molecules with the ability to activate two different receptors either on the same cell or on different cell types. To achieve prolonged plasma half-life and extended drug disposition, we have developed novel dual-acting molecules by fusing single-chain ligands of the TNFSF to heterodimerizing Fc chains (scDuokine-Fc, scDk-Fc). Incorporating costimulatory ligands of the TNF superfamily into a scDk-Fc molecule resulted in enhanced T-cell proliferation translating in an increased anti-tumor activity in combination with a primary T-cell-activating bispecific antibody. Our data show that the scDk-Fc molecules are potent immune-stimulatory molecules that are able to enhance T-cell mediated anti-tumor responses.


Pitting of malaria parasites in microfluidic devices mimicking spleen interendothelial slits.
The spleen is a hematopoietic organ that participates in cellular and humoral immunity. It also serves as a quality control mechanism for removing senescent and/or poorly deformable red blood cells (RBCs) from circulation. Pitting is a specialized process by which the spleen extracts particles, including malaria parasites, from within circulating RBCs during their passage through the interendothelial slits (IES) in the splenic cords. To study this physiological function in vitro, we have developed two microfluidic devices modeling the IES, according to the hypothesis that at a certain range of mechanical stress on the RBC, regulated through both slit size and blood flow, would force it undergo the pitting process without affecting the cell integrity. To prove its functionality in replicating pitting of malaria parasites, we have performed a characterization of P. falciparum-infected RBCs (P.f.-RBCs) after their passage through the devices, determining hemolysis and the proportion of once-infected RBCs (O-iRBCs), defined by the presence of a parasite antigen and absence of DAPI staining of parasite DNA using a flow cytometry-based approach. The passage of P.f.-RBCs through the devices at the physiological flow rate did not affect cell integrity and resulted in an increase of the frequency of O-iRBCs. Both microfluidic device models were capable to replicate the pitting of P.f.-RBCs ex vivo by means of mechanical constraints without cellular involvement, shedding new insights on the role of the spleen in the pathophysiology of malaria.


The role of cholesterol in invasion and growth of malaria parasites.
Malaria parasites are unicellular eukaryotic pathogens that develop through a complex lifecycle involving two hosts, an anopheline mosquito and a vertebrate host. Throughout this lifecycle, the parasite encounters widely differing conditions and survives in distinct ways, from an intracellular lifestyle in the vertebrate host to exclusively extracellular stages in the mosquito. Although the parasite relies on cholesterol for its growth, the parasite has an ambiguous relationship with cholesterol: cholesterol is required for invasion of host cells by the parasite, including hepatocytes and erythrocytes, and for the development of the parasites in those cells. However, the parasite is unable to produce cholesterol itself and appears to remove cholesterol actively from its own plasma membrane, thereby setting up a cholesterol gradient inside the infected host erythrocyte. Overall a picture emerges in which the parasite relies on host cholesterol and carefully controls its transport. Here, we describe the role of cholesterol at the different lifecycle stages of the parasites.


Monoclonal antibody to the murine type 3 complement receptor inhibits adhesion of myelomonocytic cells in vitro and inflammatory cell recruitment in vivo.
Macrophage interactions with extracellular matrix and other cells are important in phagocytosis, inflammation, and immunity. To learn more about the surface molecules involved in adhesion we compared the binding of murine macrophages and polymorphonuclear leukocytes (PMN) with artificial substrate in vitro. A distinctive type of adhesion of thioglycollate-elicited peritoneal macrophages (TPM) to bacteriologic plastic (BP) was defined, which was pronase-sensitive, Mg2+-dependent, and required cytoskeletal stabilization. A rat mAb designated 5C6 was isolated because it inhibited TPM attachment to BP, as well as mediating detachment of TPM adherent to that substratum. In addition, it inhibited the attachment of PMN to tissue culture plastic. This antiadhesive property of 5C6 mAb required intact IgG; the F(ab')2 fragment was partially effective and Fab was ineffective. 5C6 recognized the type 3 complement receptor, inhibiting rosetting of EAC3bi to TPM and immunoprecipitating a heterodimer of 160 and 95 kD that comigrated with the M1/70 immunoprecipitate. 5C6 recognized a pronase-stable epitope distinct from that of M1/70. Other mAbs, including M1/70 (CR3) and 2.4G2 (FcR), failed to have any antiadhesive effect in vitro. The inhibitory activity of 5C6 in short-term adhesion assays correlated with its inhibition of recruitment of myelomonocytic cells to a thioglycollate-elicited peritoneal exudate in vivo, after intravenous injection of mAb. 5C6 IgG inhibited recruitment of myelomonocytic cells by 84 +/- 3% at 1 d compared with saline-injected controls. The F(ab')2 fragment and a class-matched control IgG had little effect. Recruitment of TPM at 4 d was also efficiently inhibited by 5C6 IgG. 5C6 IgG was not cytotoxic, had no effect on marrow egress, did not cause increased phagocytic clearance of circulating neutrophils, and had no adverse effect on chemotaxis in vitro. We show that CR3 alone of the LFA-family is necessary for the recruitment of myelomonocytic cells to inflammatory stimuli such as thioglycollate broth. This strategy may be of general use in isolating reagents that inhibit the adhesive function of CR3 and provides a novel approach to antiinflammatory therapy.


Mechanical support of the circulation followed by cardiac transplantation.
Improvements in both mechanical circulatory support devices and immune therapy promise a wider use of sequential mechanical support as a bridge to orthotopic cardiac transplantation. The intra-aortic balloon pump, the left and right ventricular assist pumps, and the pneumatic artificial heart represent the range of devices capable of keeping a patient alive who is awaiting a donor organ. The major difficulty in using circulatory support devices is infection, which is caused by their required percutaneous tubes. We report here our experiences with mechanical circulatory support devices as a bridge to cardiac transplantation. In a series of 31 consecutive transplant procedures, six patients have required preoperative mechanical circulatory support. The intra-aortic balloon pump was used in two patients for 2 and 14 days, respectively, before transplantation. Both patients are well 10 and 11 months after the transplant procedure. Two patients required the left ventricular assist device for 11 and 21 days and are alive 3 weeks and 8 months, respectively, after transplantation. One patient was supported by the pneumatic artificial heart for 10 days before a donor heart became available but died of septic shock 17 days after transplantation. A second patient received a pneumatic artificial heart 7 days after transplantation when the heart transplant failed. He has been in stable condition for 45 days but is recovering from renal failure. Our early experiences indicate that either partial or total mechanical support as a bridge to transplantation is successful if overwhelming sepsis or renal failure can be avoided.


[Retinal complications of pars plana vitrectomy].
A series of 500 consecutive pars plana vitrectomies was evaluated with regard to retinal complications. If the retina is attached preoperatively the risk of retinal complications is only 2.0%; however, in cases with preoperative detachment it rises to 9.2%. When membrane peeling is performed the risk rises by a further 7.3%. These statistics support the view that early vitrectomy is today a safe operation in cases with a simple initial anatomical situation. In contrast, late vitrectomy in cases where the initial anatomical situation is complex and membrane-peeling has to be performed is necessarily associated with a higher rate of complications.


Optimization of enzyme-based assays in coagulation testing.
Optimized assays for antithrombin III and plasminogen have been developed based on a study of the kinetic parameters Km and Kcat for four commercially available substrates: the p-nitroanilide derivatives of D-Phe-pipecolyl-Arg (S-2238), and toluenesulfonyl-Gly-Pro-Arg (Chromozym TH), which are thrombin substrates; D-Val-Leu-Lys (S-2251), a plasminogen/streptokinase substrate; and alpha-N-carbobenzoxy-L-lysine thiobenzyl ester, a substrate for both enzymes. We used a centrifugal analyzer system for rapid data acquisition and interactive analysis. Optimized conditions for assay of a particular enzyme are not constant for different substrates in the same buffering agent. For example, in 1,4-piperazine diethanesulfonic acid buffer at 37 degrees C, thrombin-catalyzed hydrolysis of Chromozym TH is optimal at 125 mmol/L buffer, 100 mmol/L NaCl, and pH 8.2, whereas substitution of S-2238, also a tripeptide p-nitroanilide, yields optimal hydrolysis at 85 mmol/L buffer, 300 mmol/L NaCl, and pH 7.2. We conclude that optimized assay conditions are best obtained by an extensive survey of available buffers and a detailed investigation of the effects of variation in pH and in the concentrations of the buffer and auxiliary reagents through use of both one-factor-at-a-time and multivariate response surface experimentation.


Activity and safety of crizotinib in patients with advanced clear-cell sarcoma with MET alterations: European Organization for Research and Treatment of Cancer phase II trial 90101 'CREATE'.
Clear-cell sarcoma (CCSA) is an orphan malignancy, characterized by a specific t(12;22) translocation, leading to rearrangement of the EWSR1 gene and overexpression of MET. We prospectively investigated the efficacy and safety of the tyrosine kinase inhibitor crizotinib in patients with advanced or metastatic CCSA. Patients with CCSA received oral crizotinib 250 mg twice daily. Primary end point was objective response rate (ORR), secondary end points included duration of response, disease control rate (DCR), progression-free survival (PFS), progression-free rate (PFR), overall survival (OS), OS rate and safety. The study design focused on MET+ disease with documented rearrangement of the EWSR1 gene by fluorescence in situ hybridization. Among 43 consenting patients with the local diagnosis of CCSA, 36 had centrally confirmed CCSA, 28 of whom were eligible, treated and assessable. Twenty-six out of the 28 patients had MET+ disease, of whom one achieved a confirmed partial response and 17 had stable disease (SD) (ORR 3.8%, 95% confidence interval: 0.1-19.6). Further efficacy end points in MET+ CCSA were DCR: 69.2% (48.2% to 85.7%), median PFS: 131 days (49-235), median OS: 277 days (232-442). The 3-, 6-, 12- and 24-month PFR was 53.8% (34.6-73.0), 26.9% (9.8-43.9), 7.7% (1.3-21.7) and 7.7% (1.3-21.7), respectively. Among two assessable MET- patients, one had stable disease and one had progression. The most common treatment-related adverse events were nausea [18/34 (52.9%)], fatigue [17/34 (50.0%)], vomiting [12/34 (35.3%)], diarrhoea [11/34 (32.4%)], constipation [9/34 (26.5%)] and blurred vision [7/34 (20.6%)]. The PFS with crizotinib in MET+ CCSA is similar to results achieved first-line in non-selected metastatic soft tissue sarcomas with single-agent doxorubicin. The PFS is similar to results achieved with pazopanib in previously treated sarcoma patients. EORTC 90101, EudraCT number 2011-001988-52, NCT01524926.


Computed tomography fluoroscopy versus conventional computed tomography guidance for biopsy of intrathoracic lesions: a retrospective review of 1143 consecutive procedures.
To compare the success rates, complication rates, and radiation doses of conventional computed tomography (CCT) versus computed tomography with fluoroscopy (CTF) during image-guided percutaneous needle biopsies of intrathoracic lesions. We conducted a retrospective analysis of 1143 consecutive intrathoracic biopsies performed under computed tomography guidance at a single tertiary care center. For each procedure we recorded patient age and sex, lesion size and location, and radiation dose administered to the patient. Thereafter, appropriate tests of statistical significance were applied to compare rates of success, complications, and radiation between CCT and CTF guidance. After correcting for lesion size, CTF was associated with an odds ratio (OR) of 6.07 [95% confidence interval (CI): 2.23-16.50] for technical procedural success, an OR of 0.79 (95% CI: 0.55-1.15) for fine-needle aspiration success, an OR of 2.11 (95% CI: 1.02-4.38) for core-needle biopsy success, and an OR of 1.45 (95% CI: 1.00-2.21) for overall success when compared with CCT. CTF was associated with an OR of 1.10 (95% CI: 0.35-3.48) for complications when compared with CCT. Mean dose-length product per procedure was 1332 mGy cm for CCT and 1730 mGy cm for CTF (P=0.027). CCT and CTF are both valuable tools for intrathoracic biopsy. CTF improves technical and core-needle biopsy success at the expense of higher radiation doses to the patient. Operator confidence with CCT and CTF in light of lesion characteristics should guide the choice to select one over the other.


Messenger RNA decay and maturation in Bacillus subtilis.
Our understanding of the ribonucleases that act to process and turn over RNA in Bacillus subtilis, a model Gram-positive organism, has increased greatly in recent years. This chapter discusses characteristics of B. subtilis ribonucleases that have been shown to participate in messenger RNA maturation and decay. Distinct features of a recently discovered ribonuclease, RNase J1, are reviewed, and are put in the context of a mechanism for the mRNA decay process in B. subtilis that differs greatly from the classical model developed for E. coli. This chapter is divided according to three parts of an mRNA-5' end, body, and 3' end-that could theoretically serve as sites for initiation of decay. How 5'-proximal elements affect mRNA half-life, and especially how these elements interface with RNase J1, forms the basis for a set of "rules" that may be useful in predicting mRNA stability.


MR-less surface-based amyloid assessment based on 11C PiB PET.
β-amyloid (Aβ) plaques in brain's grey matter (GM) are one of the pathological hallmarks of Alzheimer's disease (AD), and can be imaged in vivo using Positron Emission Tomography (PET) with (11)C or (18)F radiotracers. Estimating Aβ burden in cortical GM has been shown to improve diagnosis and monitoring of AD. However, lacking structural information in PET images requires such assessments to be performed with anatomical MRI scans, which may not be available at different clinical settings or being contraindicated for particular reasons. This study aimed to develop an MR-less Aβ imaging quantification method that requires only PET images for reliable Aβ burden estimations. The proposed method has been developed using a multi-atlas based approach on (11)C-PiB scans from 143 subjects (75 PiB+ and 68 PiB- subjects) in AIBL study. A subset of 20 subjects (PET and MRI) were used as atlases: 1) MRI images were co-registered with tissue segmentation; 2) 3D surface at the GM-WM interfacing was extracted and registered to a canonical space; 3) Mean PiB retention within GM was estimated and mapped to the surface. For other participants, each atlas PET image (and surface) was registered to the subject's PET image for PiB estimation within GM. The results are combined by subject-specific atlas selection and Bayesian fusion to generate estimated surface values. All PiB+ subjects (N = 75) were highly correlated between the MR-dependent and the PET-only methods with Intraclass Correlation (ICC) of 0.94, and an average relative difference error of 13% (or 0.23 SUVR) per surface vertex. All PiB- subjects (N = 68) revealed visually akin patterns with a relative difference error of 16% (or 0.19 SUVR) per surface vertex. The demonstrated accuracy suggests that the proposed method could be an effective clinical inspection tool for Aβ imaging scans when MRI images are unavailable.


Reducing future nutrient inputs to the Black Sea.
Rivers export increasing amounts of dissolved inorganic (DIN, DIP) and organic (DON, DOP) nitrogen and phosphorus to the Black Sea causing coastal eutrophication. The aim of this study is to explore future trends in river export of these nutrients to the sea through a sensitivity analysis. We used the Global NEWS (Nutrient Export from WaterSheds) model to this end. We calculated that between 2000 and 2050 nutrient inputs to the Black Sea may increase or decrease, depending on the assumed environmental management. We analyzed the effects of agricultural and sewage management on nutrient inputs to the sea in 2050 relative to two Millennium Ecosystem Assessment (MEA) scenarios, Global Orchestration (GO) and Adaptive Mosaic (AM). In these baselines, total N and P inputs to the Black Sea decrease between 2000 and 2050, but not for all rivers and nutrient forms. Our results indicate that it is possible to reduce nutrient inputs to the sea further between 2000 and 2050 in particular for dissolved inorganic N and P and for many river basins, but not for all. For scenarios assuming combined agricultural and sewage management dissolved inorganic N and P inputs to the Black Sea are reduced by up to two-thirds between 2000 and 2050 and dissolved organic N and P inputs by one-third. River export of DIN is mainly affected by agricultural management and that of DIP by sewage management. On the other hand, in scenarios assuming increased fertilizer use for, for instance bioenergy crops, nutrient inputs to the sea increase. An increase in DIP inputs by southern rivers seems difficult to avoid because of the increasing number of people connected to sewage systems.


The application of monolithic columns in pesticides analysis.
HPLC and HPLC/MS are the most widely used analytical techniques in the field of pesticides analysis. In recent years, there has been considerable focus on fast separations in HPLC in order to reduce analysis time as well as cost. Monolithic columns, consisting of continuous beds with macropores and mesopores, can meet this requirement and have been widely used in the medical and biological fields. However, it has seldom been used when analyzing pesticides. In this work, the application of monolithic columns in pesticides analysis and their advantages are evaluated and compared with those obtained using conventional packed columns.


Case reports. 1. An autopsy case of fatal arrhythmia induced by injuries of the atrioventricular conduction system: a case report.
A 65-year-old woman died three days after being involved in a traffic accident, following an episode of ventricular fibrillation. She was diagnosed as having suffered cardiac contusion, liver contusion, mediastinal hematoma and rib fracture on admission. Her electrocardiogram showed complete right bundle branch block, complete atrioventricular block, and right axis deviation. Aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase and creatine kinase-MB were found to be elevated on biochemical blood analysis. These findings recovered and her condition appeared to improve daily. At autopsy, epicardial and intramyocardial haemorrhage were macroscopically seen in the posterior wall of the bilateral ventricles. On microscopic examination, there was evidence of fresh haemorrhage and coagulative necrosis with inflammatory reaction in the ordinary myocardium and adipose tissue around the atrioventricular node, which had spread to the proximal portion of the His' bundle. It is considered that these findings caused ventricular fibrillation to occur, and that the cause of death in this case was myocardial contusion due to blunt thoracic injury. This case would indicate that myocardium nearby atrioventricular junction is vulnerable to external force. Moreover, it would seem that fatal arrhythmia occasionally occurs during the follow-up stage, despite the lack of any significant clinical findings.


Association between biological markers of sickle cell disease and cognitive functioning amongst Cameroonian children.
Some of the major complications of sickle cell disease (SCD) occur in the brain and apart from overt stroke, patients also present with cognitive impairments. We sought to evaluate the prevalence of cognitive deficits as well as their biological predicting factors in young SCD patients in Cameroon. The cognitive performances of Cameroonian SCD young patients were evaluated using a neuropsychological test battery assessing four domains of cognitive functioning (executive functions, attention, memory, and sensory-motor skills) previously adapted and normalized on healthy subjects in Yaoundé. Up to 37.5% of the 96 SCD patients aged 6 to 24 years (M = 13.5, SD = 4.9) had mild-to-severe cognitive deficits. The cognitive deficits tend to increase with age. There was a significant effect of SCD on executive functions and attention, whereas SCD patients performed as well as controls on memory and sensory-motor skills tests. Structural equation models showed a significant association between (a) severe anemia and lower executive functioning, (b) low fetal hemoglobin levels and lower executive functioning and attention, (c) history of cerebrovascular accidents and lower performances in executive functioning, sensory-motor skills, and memory, (d) pathological electroencephalogram and lower attention, and (e) abnormal Transcranial Doppler and lower memory. SCD patients in Cameroon presented a very high prevalence of cognitive deficits, with a specific impairment of executive functions and attention. Routine neuropsychological evaluation for early detection of cognitive deficits in SCD patients could represent a cost-effective tool to implement in resource-limited contexts such as in sub-Saharan Africa.


One-year effectiveness and safety of ustekinumab in ulcerative colitis: a multicenter real-world study from Italy.
Efficacy and safety of ustekinumab for the treatment of ulcerative colitis (UC) has been demonstrated in clinical trials, but few real-world data are available so far. The aim of this study was to assess effectiveness and safety of ustekinumab in a cohort of refractory UC patients. Data of patients with moderate to severe UC treated with ustekinumab were retrospectively collected. Primary endpoint was steroid-free clinical remission at weeks 24 and 52 of therapy. Secondary endpoints were treatment response, endoscopic remission, treatment persistence at 12 months and safety. A total of 68 patients [males 63%; median (range) age 42 (16-72) years] were included. Almost all patients (97%) were biologics experienced. At weeks 24 and 52, 31% and 50% of patients achieved steroid-free clinical remission, 84% and 82% had clinical response, respectively. At the end of follow-up, there was a significant reduction of pMS from baseline (p < 0.001) and of steroid use (p < 0.001). At week 52, 22% of the available endoscopies (18/38) showed mucosal healing. The probability to persist in therapy at week 52 was 87%. Only one adverse event occurred. Data from our real-life cohort of refractory UC patients suggest satisfactory effectiveness and a good safety of ustekinumab.


Can the Pain Attitudes and Beliefs Scales be adapted for use in the context of osteoarthritis with general practitioners and physiotherapists?
Conservative, first-line treatments (exercise, education and weight-loss if appropriate) for hip and knee joint osteoarthritis are underused despite the known benefits. Clinicians' beliefs can affect the advice and education given to patients, in turn, this can influence the uptake of treatment. In New Zealand, most conservative OA management is prescribed by general practitioners (GPs; primary care physicians) and physiotherapists. Few questionnaires have been designed to measure GPs' and physiotherapists' osteoarthritis-related health, illness and treatment beliefs. This study aimed to identify if a questionnaire about low back pain beliefs, the Pain Attitudes and Beliefs Scale for Physiotherapists (PABS-PT), can be adapted to assess GP and physiotherapists' beliefs about osteoarthritis. This study used a cross-sectional observational design. Data were collected anonymously from GPs and physiotherapists using an online survey. The survey included a study-specific demographic and occupational characteristics questionnaire and the PABS-PT questionnaire adapted for osteoarthritis. All data were analysed using descriptive statistics, and the PABS-PT data underwent principal factor analysis. In total, 295 clinicians (87 GPs, 208 physiotherapists) participated in this study. The principal factor analysis identified two factors or subscales (categorised as biomedical and behavioural), with a Cronbach's alpha of 0.84 and 0.44, respectively. The biomedical subscale of the PABS-PT appears appropriate for adaptation for use in the context of osteoarthritis, but the low internal consistency of the behavioural subscale suggests this subscale is not currently suitable. Future research should consider the inclusion of additional items to the behavioural subscale to improve internal consistency or look to develop a new, osteoarthritis-specific questionnaire. This trial was part of the primary author's PhD, which began in 2012 and therefore this study was not registered.


Application of the enzyme-linked immunosorbent assay for the detection of platelet antibodies in dogs.
An enzyme-linked immunosorbent assay (ELISA) was developed to detect serum antibodies reactive to canine platelets. The method used an anti-canine immunoglobulin G horseradish peroxidase conjugate, O-dianisidine, as a water-soluble chromagen, and pooled platelets as a temporary antibody support. The ELISA was compared with a conventional immunoinjury method measuring the accelerating effect of platelet factor 3 on coagulation. The ELISA was more sensitive in the detection of antiplatelet antibodies in canine patients with idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, and systemic lupus erythematosus. Serial dilution of rabbit anti-canine platelet serum revealed a positive correlation between the strength of the antiserum and the degree of color measured spectophotometrically, indicating that the absorbance value may be useful in estimating the amount of antibody present in the serum in indirect tests and on the platelets in direct tests.


Potential Deoxycytidine Kinase Inhibitory Activity of Amaryllidaceae Alkaloids: An In Silico Approach.
Plants of the Amaryllidaceae family have been under intense scrutiny for the presence of a couple of alkaloidal secondary metabolites with endued cytotoxic activity, such as pancratistatin (1), 7-deoxypancratistatin (2), narciclasine (3), 7-deoxynarciclasine (4), trans-dihydronarciclasine (5), and 7-deoxy-trans-dihydronarciclasine (6). Nevertheless, preclinical evaluation of these alkaloids has been put on hold because of the limited quantity of materials available from isolation. To explore the underlying cytotoxic molecular mechanisms of the Amaryllidaceae alkaloids (1-6) and to assess their absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiles using chemoinformatic tools. AutoDock 4.0 software along with different in silico chemoinformatic tools, namely PharmMapper, Molinspiration, MetaPrint2D, and admetSAR servers, were used to assess the drugability of the Amaryllidaceae alkaloids (1-6). Deoxycytidine kinase (dCK) (PDB: 1P60) was predicted as a potential target with fitting score of 5.574. In silico molecular docking of (1-6) into dCK revealed good interactions, where interesting hydrogen bonds were observed with the amino acid residues-Gly-28 and Ser-35-located in the highly conserved P-loop motif. This motif plays a special role in dCK function. Contrary to (1), in silico pharmacokinetic results have shown good absorption and permeation and thus good oral bioavailability for (2-6). The in silico docking data have proposed that the reported cytotoxic activity of the Amaryllidaceae alkaloids (1-6) could be mediated through dCK inhibition. In addition, the ADMET profile of these alkaloids is promising and thus (1-6) could be candidates for future drug development.


Transplantation of embryonic spinal cord-derived neurospheres support growth of supraspinal projections and functional recovery after spinal cord injury in the neonatal rat.
Great interest exists in using cell replacement strategies to repair the damaged central nervous system. Previous studies have shown that grafting rat fetal spinal cord into neonate or adult animals after spinal cord injury leads to improved anatomic growth/plasticity and functional recovery. It is clear that fetal tissue transplants serve as a scaffold for host axon growth. In addition, embryonic Day 14 (E14) spinal cord tissue transplants are also a rich source of neural-restricted and glial-restricted progenitors. To evaluate the potential of E14 spinal cord progenitor cells, we used in vitro-expanded neurospheres derived from embryonic rat spinal cord and showed that these cells grafted into lesioned neonatal rat spinal cord can survive, migrate, and differentiate into neurons and oligodendrocytes, but rarely into astrocytes. Synapses and partially myelinated axons were detected within the transplant lesion area. Transplanted progenitor cells resulted in increased plasticity or regeneration of corticospinal and brainstem-spinal fibers as determined by anterograde and retrograde labeling. Furthermore, transplantation of these cells promoted functional recovery of locomotion and reflex responses. These data demonstrate that progenitor cells when transplanted into neonates can function in a similar capacity as transplants of solid fetal spinal cord tissue.


Optofluidic time-stretch imaging - an emerging tool for high-throughput imaging flow cytometry.
Optical imaging is arguably the most effective tool to visualize living cells with high spatiotemporal resolution and in a nearly noninvasive manner. Driven by this capability, state-of-the-art cellular assay techniques have increasingly been adopting optical imaging for classifying different cell types/stages, and thus dissecting the respective cellular functions. However, it is still a daunting task to image and characterize cell-to-cell variability within an enormous and heterogeneous population - an unmet need in single-cell analysis, which is now widely advocated in modern biology and clinical diagnostics. The challenge stems from the fact that current optical imaging technologies still lack the practical speed and sensitivity for measuring thousands to millions of cells down to the single-cell precision. Adopting the wisdom in high-speed fiber-optics communication, optical time-stretch imaging has emerged as a completely new optical imaging concept which is now proven for ultrahigh-throughput optofluidic single-cell imaging, at least 1-2 orders-of-magnitude higher (up to ∼100 000 cells per second) compared to the existing imaging flow cytometers. It also uniquely enables quantification of intrinsic biophysical markers of individual cells - a largely unexploited class of single-cell signatures that is known to be correlated with the overwhelmingly investigated biochemical markers. With the aim of reaching a wider spectrum of experts specializing in cellular assay developments and applications, this paper highlights the essential basics of optical time-stretch imaging, followed by reviewing the recent developments and applications of optofluidic time-stretch imaging. We will also discuss the current challenges of this technology, in terms of providing new insights in basic biology and enriching the clinical diagnostic toolsets.


GASAT: a genetic local search algorithm for the satisfiability problem.
This paper presents GASAT, a hybrid algorithm for the satisfiability problem (SAT). The main feature of GASAT is that it includes a recombination stage based on a specific crossover and a tabu search stage. We have conducted experiments to evaluate the different components of GASAT and to compare its overall performance with state-of-the-art SAT algorithms. These experiments show that GASAT provides very competitive results.


Bedside ultrasound in the diagnosis of nonalcoholic fatty liver disease.
Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in the United States. While the American Association for the Study of Liver Diseases guidelines define NAFLD as hepatic steatosis detected either on histology or imaging without a secondary cause of abnormal hepatic fat accumulation, no imaging modality is recommended as standard of care for screening or diagnosis. Bedside ultrasound has been evaluated as a non-invasive method of diagnosing NAFLD with the presence of characteristic sonographic findings. Prior studies suggest characteristic sonographic findings for NAFLD include bright hepatic echoes, increased hepatorenal echogenicity, vascular blurring of portal or hepatic vein and subcutaneous tissue thickness. These sonographic characteristics have not been shown to aid bedside clinicians easily identify potential cases of NAFLD. While sonographic findings such as attenuation of image, diffuse echogenicity, uniform heterogeneous liver, thick subcutaneous depth, and enlarged liver filling of the entire field could be identified by clinicians from bedside ultrasound. The accessibility, ease of use, and low-side effect profile of ultrasound make bedside ultrasound an appealing imaging modality in the detection of hepatic steatosis. When used with appropriate clinical risk factors and steatosis involves greater than 33% of the liver, ultrasound can reliably diagnose NAFLD. Despite the ability of ultrasound in detecting moderate hepatic steatosis, it cannot replace liver biopsy in staging the degree of fibrosis. The purpose of this review is to examine the diagnostic accuracy, utility, and limitations of ultrasound in the diagnosis of NAFLD and its potential use by clinicians in routine practices.


Efficacy of Bromelain along with Trypsin, Rutoside Trihydrate Enzymes and Diclofenac Sodium Combination Therapy for the treatment of TMJ Osteoarthritis - A Randomised Clinical Trial.
Osteoarthritis (OA) is a degenerative disorder characterized by chronic inflammatory response of cartilage and articular surface involving Temporomandibular Joint (TMJ). Pain as one of the major symptom of osteoarthritis affects the quality of life and is usually managed by Non Steroidal Anti Inflammatory Drugs (NSAIDs) such as diclofenac sodium. Bromelain, trypsin and rutoside trihydrate formulation can be used to treat this disease because of anti-inflammatory and anti-oxidant effects. To assess the effectiveness of oral bromelain, trypsin, rutoside trihydrate enzymes and diclofenac sodium combination therapy over diclofenac sodium for the treatment of TMJ osteoarthritis. Thirty Patients with symptomatic TMJ osteoarthritis were randomly divided into three groups. 10 patients were treated with diclofenac sodium (Group 1), 10 were given oral enzymes (bromelain, trypsin, rutoside trihydrate) and diclofenac sodium combination (Group 2), and 10 were treated with oral enzyme preparation (bromelain, trypsin, rutoside trihydrate) (Group 3). Patients were evaluated on day 1, day 4, day 7 and day 10. Comparison of pain rating within three groups was assessed using numeric rating scale. The efficacy criteria were analysed applying ANOVA followed by post-hoc test. Inter group comparison of the effectiveness of management of pain, resulted in a value p < 0.05 between Group 2 and other two groups, which indicated that Group 2 patients responded better than Group 1 and Group 3, while p > 0.05 between Group 1 and Group 3 showed both groups responded equally to the treatment. The trial showed significant improvement in reducing pain in patients treated with oral enzymes and diclofenac sodium combination therapy.


Changes in H+-translocating vacuolar-type ATPase in the anterior silk gland cell of Bombyx mori during metamorphosis.
A proton-translocating vacuolar-type ATPase (V-ATPase) was identified and characterized in the anterior silk gland of Bombyx mori. By incubating the intact tissue with the fluorescent dye Acridine Orange, the acidified compartment was detected at the apical pole of the epithelial cells. This was observed throughout the feeding period of the fifth-instar larva until the onset of spinning. Acidification was prevented completely and reversibly by 0.8 micromol l-1 bafilomycin A1, a specific inhibitor of V-ATPase. The presence of V-ATPase in a microsomal fraction was verified by immunoblots using an antiserum to the V-ATPase holoenzyme from Manduca sexta midgut. The antiserum localized the V-ATPase to the apical plasma membrane of the anterior silk gland cells, suggesting that the enzyme is functionally active in pumping protons out of the cell towards the glandular lumen of feeding silkworm larvae. In spinning larvae, the acidification produced by the V-ATPase appears to cease, because acidic compartments were seen rarely and only in the periphery of basal cytoplasm, and because immunocytochemical staining for the V-ATPase was greatly reduced at the apical surface. The metamorphic changes in relation to the occurrence of V-ATPase corresponded well with the ultrastructural changes in the anterior silk gland cell of Bombyx mori larvae.