Patch recordings from the electrocytes of Electrophorus electricus. Na currents and PNa/PK variability.
Sodium currents were recorded in cell-attached and inside-out patches from the innervated membrane of Electrophorus electrocytes. Electrocytes from Sachs and main electric organs were prepared as described by Pasquale et al. (1986. J. Membr. Biol. 93:195.). Maximal currents in the Sachs organ, measured with 1-2 microns diameter patch pipettes and at room temperature, were in the range of 20 to 300 pA (27 patches) and were obtained near +10 mV. This range of current corresponds to approximately 70 to 1,300 channels in a patch. Maximal current in main organ cells also occurred near +10 mV and were in the range of 100 to 400 pA. Delayed K current was observed in a few patches. The inactivation phase of the currents during maintained depolarizations appears to be a single-exponential relaxation. The time constant decreases from 1 ms near -55 mV to a minimum of 0.3 ms near 0 mV, and then gradually increases with stronger depolarization. The mean currents are half inactivated near -90 mV with an apparent voltage dependence of e-fold per 6 mV. No apparent differences were observed in the decay time course or steady-state inactivation of the currents in the same patch before and after excision. From ensemble fluctuation analysis the peak open probability was found to be approximately 0.5 at +25 mV and increased only gradually with larger depolarizations. The single channel conductances were approximately 20 pS with 200 mM Na outside and 200 mM K inside, and 40 pS in 400 mM solutions. Reversal potentials in the 200 Na parallel 200 K solutions ranged from +51 to +94 mV in multichannel patches, corresponding to selectivity ratios PNa/PK from 8 to 43. Large differences in reversal potentials were seen even among patches from the same cell. Several controls rule out obvious sources of error in the reversal potential measurements. It is concluded that there is heterogeneity in the selectivity properties of the Na channels.


Lack of association of herpes simplex virus type 2 seropositivity with the progression of HIV infection in the HERS cohort.
Many studies have chronicled the "epidemiologic synergy" between human immunodeficiency virus (HIV) and herpes simplex virus type 2 (HSV-2). HIV adversely affects the natural history of HSV-2 and results in more frequent and severe HSV-2 reactivation. Few longitudinal studies, however, have examined whether HSV-2 is associated with increased HIV plasma viral loads or decreased CD4 counts. The authors estimated the effect of HSV-2 seropositivity on HIV RNA viral load and on CD4 count over time among 777 HIV-seropositive US women not receiving suppressive HSV-2 therapy in the HIV Epidemiology Research Study (1993-2000). Linear mixed models were used to assess the effect of HSV-2 on log HIV viral load and CD4 count/mm(3) prior to widespread initiation of highly active antiretroviral therapy. Coinfection with HSV-2 was not associated with HIV RNA plasma viral loads during study follow-up. There was a statistically significant association between HSV-2 seropositivity and CD4 count over time, but this difference was small and counterintuitive at an increase of 8 cells/mm(3) (95% confidence interval: 2, 14) per year among HSV-2-seropositive women compared with HSV-2-seronegative women. These data do not support a clinically meaningful effect of baseline HSV-2 seropositivity on the trajectories of HIV plasma viral loads or CD4 counts.


The pursuit of equity in NHS resource allocation: should morbidity replace utilisation as the basis for setting health care capitations?
Although the English NHS has been described as a world leader in pioneering methods of distributing expenditure in relation to population needs, concerns about the legitimacy of using the current utilisation-based model to allocate health service resources are mounting. In this paper, we present a critical review of NHS resource allocation in England and demonstrate the feasibility and impact of using direct health estimates as a basis for setting health care capitations. Comparing target allocations for the inpatient treatment of coronary heart disease in a sample of 34 primary care trusts in contrasting locations in England, we find that a morbidity-based model would result in a significant shift in hospital resources away from deprived areas, towards areas with older demographic profiles and towards rural areas. Discussing the findings in relation to a wider policy context that is generally concerned to direct more health care resources towards the poor, the paper concludes by calling for greater clarity between the goals of health care equity and health equity. Whilst the former demands that the legitimate needs of demographically older populations for more health care resources are acknowledged, the goal of health equity requires real political commitment to resource broader social policy initiatives.


Efficacy and safety of first-line anlotinib-based combinations for advanced non-small cell lung cancer: a three-armed prospective study.
The evidence of combined therapies of multi-target agents in first-line treatment of advanced non-small cell lung cancer (NSCLC) was limited. This study aimed to evaluate the safety and efficacy of anlotinib combined with epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), chemotherapy, or immune checkpoint inhibitor (ICI) in advanced NSCLC. This open-label, three-arm, prospective study (NCT03628521) enrolled untreated locally advanced/metastatic NSCLC patients. Patients with EGFR mutation NSCLC received anlotinib and erlotinib (cohort A). Patients without EGFR/ALK/ROS1 mutation received anlotinib combined with carboplatin plus pemetrexed/gemcitabine (cohort B), or sintilimab (cohort C). The primary outcomes were safety and objective response rate (ORR). The secondary endpoints included progression-free survival (PFS), disease control rate (DCR), and overall survival (OS). Treatments were performed for at least 2 cycles and efficacy was evaluated every 2 cycles using RECIST version 1.1. Safety was assessed throughout the study. A total of 30, 30, and 22 patients were enrolled in cohorts A, B, and C, respectively. There were 3 patients did not complete the treatment in cohort A. In cohorts A and B, ≥ grade 3 treatment-related adverse events (TRAEs) occurred in 77.3% and 60.0% of patients, respectively. The most common TRAEs were rash (10.0%) and decreased platelet count (30.0%) in cohorts A and B, respectively. The ORRs were 92.9% and 60.0% in cohorts A and B, respectively, and DCRs were 96.4% and 96.7%, respectively. The ORR and incidence of ≥ grade 3 TRAEs of cohort C were, which 72.7% and 54.5%, which had been published previously. Median PFSs [95% confidence interval (CI)] were 21.6 (15.6 to 24.9), 13.0 [10.5 to not estimated (NE)], and 15.6 (12.9 to NE) months in cohorts A, B, and C, respectively. Median OS was 28.1 (95% CI: 21.82 to NE) months in cohort B. The 24-month OS rates in cohorts A and C were 87.1% and 83.9%, respectively. Anlotinib-based combinations with EGFR-TKI, chemotherapy, and ICI are well-tolerated and encouraging as first-line therapies for advanced NSCLC, which could be verified in future studies. Anlotinib-based combination might provide multiple choices for first-line treatment in patients with advanced NSCLC.


Sociospatial Disparities in "Third Place" Availability in the United States.
Tertiary to home and work, "third places" serve as opportunity structures that transmit information and facilitate social capital and upward mobility. However, third places may be inequitably distributed, thereby exacerbating disparities in social capital and mobility. The authors use tract-level data from the National Neighborhood Data Archive to examine the distribution of third places across the United States. There were significant disparities in the availability of third places. Higher poverty rates were associated with fewer third places. Tracts with the smallest shares of Black and Hispanic populations had comparatively more third places. However, this racial disadvantage was not linear, suggesting potential buffering effects in places with the largest shares of Black and Hispanic populations. There was also a rural disadvantage, except in the most isolated rural tracts. This study demonstrates the value of conceptualizing and measuring third places to understand sociospatial disparities in the availability of these understudied opportunity structures.


Molecular characterization and epidemiology of Toxoplasma gondii isolates from free-range chickens in the southwest region of Goiás: new genotypes.
The purpose of this study was to isolate Toxoplasma gondii from tissues of free-range chickens in the southwestern region of Goiás, to detect and molecularly characterize the genetic material of the parasite, and to determine the seroprevalence of the protozoan parasite in these animals. A seroprevalence of T. gondii antibodies of 76% (19/25) was found among the chickens, while genetic material from their tissues was detected in 56% (14/25). A total of 14 isolates was obtained in the bioassay, ten of which were considered acute, eight were considered isolates of high virulence lethal to mice, and four of low virulence, considered non-lethal but with the ability to chronify the infection. Seven of the ten isolates showed significant morphometric differences from the RH strain, in terms of nucleus-complex-apical distance, length and width. Genotyping of the acute isolates was performed by RFLP-PCR, using 11 genetic markers: SAG1, SAG2 (3'SAG2 and 5'SAG2), alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and APICO. The results were compared and classified according to the genotypes listed on the ToxoDB Platform, where different profiles were observed indicating the presence of two known genotypes (#7 and #63) and five new genotypes (NEW 3, NEW4, NEW5, NEW6, NEW 7). The results showed high seroprevalence, isolation rate, molecular detection and genotypic variations of T. gondii in free-range chickens in the southwestern region of Goiás.


Assessing Adherence to Responsible Reporting of Suicide Guidelines in the Canadian News Media: A 1-year Examination of Day-to-day Suicide Coverage: Évaluer la conformité au journalisme responsable en matière de directives sur le suicide dans les médias canadiens d'information: Un examen d'une année de la couverture quotidienne du suicide.
This study aims to examine routine day-to-day suicide reporting in the Canadian media, giving a descriptive overview of the tone and content of news articles. The primary objective is to assess adherence to responsible reporting of suicide recommendations in news articles about suicide. A secondary objective is to categorize these articles according to their focus. A tertiary objective is to compare guideline adherence across the different categories of articles. We collected news articles containing the keyword "suicide" from 47 Canadian news sources between April 1, 2019, and March 31, 2020. Articles were read and coded for their adherence to responsible reporting of suicide recommendations. Articles were also allotted into categories according to their focus and primary suicide discussed. Frequency counts and percentages of adherence were calculated for all key variables-both overall and by category of article. Chi-square tests were also conducted to assess for variations in adherence by category of article. The procedures resulted in 1,330 coded articles. On the one hand, there was high overall adherence to several recommendations. For example, over 80% of articles did not give a monocausal explanation, glamourize the death, appear on the front page, include sensational language, or use discouraged words. On the other hand, there was low adherence to other recommendations, especially those related to putatively protective content. For example, less than 25% included help-seeking information, quoted an expert, or included educational content. Cross-category analysis indicated that articles about events/policies/research and Indigenous people had the highest proportions of adherence, while articles about murder-suicide and high-profile suicides had the lowest adherence. While a substantial proportion of articles generally adhere to suicide reporting recommendations, several guidelines are frequently underapplied, especially those concerning putatively helpful content. This indicates room for improvement in the responsible reporting of suicide.


Exploring the cognitive and emotional correlates to proactive and reactive aggression in a sample of detained girls.
The current study examined the distinction between reactive and proactive aggression in a sample of detained girls (N = 58) aged 12 to 18. This study employed a self-report measure of aggression that was designed explicitly to assess both the forms that aggression takes (i.e., relational and overt), as well as the functions that aggression serves (i.e., reactive and proactive). Reactive aggression was uniquely associated with poorly regulated emotion and anger to perceived provocation, whereas proactive aggression was uniquely associated with callous-unemotional (CU) traits and biased outcome expectations for aggression. While overt aggression appeared to largely account for these associations, relational aggression showed strong and unique associations with CU traits. The current findings highlight the importance of assessing reactive and proactive aggression, as well as both overt and relational aggression, in detained girls.


Mechanisms of NADPH oxidase activation in human neutrophils: p67phox is required for the translocation of rac 1 but not of rac 2 from cytosol to the membranes.
NADPH oxidase is the enzyme complex responsible for the production of oxygen radicals in phagocytes. On neutrophil stimulation, the cytosolic components of NADPH oxidase, p67phox and p47phox, as well as the Ras-related G-protein rac 2, are translocated from the cytosol to cell membranes where they associate with a flavocytochrome b to form a functional complex. Besides rac 2, rac 1 G-protein is also involved in the activation of the NADPH oxidase, but, to date, it has not been documented whether it is also translocated in activated neutrophils. In this paper we show that: (a) in neutrophils stimulated with formylmethionyl-leucylphenylalanine, concanavalin A or phorbol 12-myristate 13-acetate, both rac 1 and rac 2 are translocated from cytosol to the membranes; (b) in neutrophils from a patient with a form of chronic granulomatous disease in which p67phox is absent, rac 2 and p47phox were translocated as in normal neutrophils on stimulation with the above agonists, but rac 1 failed to be translocated from the cytosol to the membranes. This is the first demonstration that, in activated neutrophils, rac 1 is translocated from the cytosol to the membranes and this translocation requires p67phox. These results, coupled with those showing that rac 2 is not translocated in activated neutrophils lacking p47phox [El Benna, Ruedi and Babior (1994) J. Biol. Chem. 269, 6729-6734], may suggest that the assembly of the cytosolic components of NADPH oxidase on the plasma membrane takes place through selective coupling of activated rac 1 and rac 2 with p67phox and p47phox respectively.


Gender and racial differences in nonalcoholic fatty liver disease.
Due to the worldwide epidemic of obesity, nonalcoholic fatty liver disease (NAFLD) has become the most common cause of elevated liver enzymes. NAFLD represents a spectrum of liver injury ranging from simple steatosis to nonalcoholic steatohepatitis (NASH) which may progress to advanced fibrosis and cirrhosis. Individuals with NAFLD, especially those with metabolic syndrome, have higher overall mortality, cardiovascular mortality, and liver-related mortality compared with the general population. According to the population-based studies, NAFLD and NASH are more prevalent in males and in Hispanics. Both the gender and racial ethnic differences in NAFLD and NASH are likely attributed to interaction between environmental, behavioral, and genetic factors. Using genome-wide association studies, several genetic variants have been identified to be associated with NAFLD/NASH. However, these variants account for only a small amount of variation in hepatic steatosis among ethnic groups and may serve as modifiers of the natural history of NAFLD. Alternatively, these variants may not be the causative variants but simply markers representing a larger body of genetic variations. In this article, we provide a concise review of the gender and racial differences in the prevalence of NAFLD and NASH in adults. We also discuss the possible mechanisms for these disparities.


Generation separation in simple structured life cycles: models and 48 years of field data on a tea tortrix moth.
Population cycles have fascinated ecologists since the early nineteenth century, and the dynamics of insect populations have been central to understanding the intrinsic and extrinsic biological processes responsible for these cycles. We analyzed an extraordinary long-term data set (every 5 days for 48 years) of a tea tortrix moth (Adoxophyes honmai) that exhibits two dominant cycles: an annual cycle with a conspicuous pattern of four or five single-generation cycles superimposed on it. General theory offers several candidate mechanisms for generation cycles. To evaluate these, we construct and parameterize a series of temperature-dependent, stage-structured models that include intraspecific competition, parasitism, mate-finding Allee effects, and adult senescence, all in the context of a seasonal environment. By comparing the observed dynamics with predictions from the models, we find that even weak larval competition in the presence of seasonal temperature forcing predicts the two cycles accurately. None of the other mechanisms predicts the dynamics. Detailed dissection of the results shows that a short reproductive life span and differential winter mortality among stages are the additional life-cycle characteristics that permit the sustained cycles. Our general modeling approach is applicable to a wide range of organisms with temperature-dependent life histories and is likely to prove particularly useful in temperate systems where insect pest outbreaks are both density and temperature dependent.


Alpha-hydroxytamoxifen is a substrate of hydroxysteroid (alcohol) sulfotransferase, resulting in tamoxifen DNA adducts.
When alpha-hydroxytamoxifen (alpha-OHTAM) was incubated with rat liver hydroxysteroid (alcohol) sulfotransferase a (STa) and 3'-phosphoadenosine 5'-phosphosulfate, (E)-alpha-OHTAM was found to be a better substrate for STa than (Z)-alpha-OHTAM. To explore the formation of tamoxifen (TAM)-derived DNA adducts, DNA was incubated with STa and either (E)-alpha-OHTAM or (Z)-alpha-OHTAM in the presence of 3'-phosphoadenosine 5'-phosphosulfate. Using 32P-postlabeling analysis, the amount of TAM-DNA adducts resulting from (E)-alpha-OHTAM was 29 times higher than that observed with (E)-alpha-OHTAM alone. Using (Z)-alpha-OHTAM and STa, some TAM-DNA adducts were also detected but at levels 6.5 times lower than that observed with (E)-alpha-OHTAM and STa. When compared with standards of stereoisomers of 2'-deoxyguanosine 3'-monophosphate-N2-tamoxifen, the major tamoxifen adduct was identified chromatographically as an epimer of the trans form of alpha-(N2-deoxyguanosinyl)tamoxifen, and the minor adduct was identified as an epimer of the cis form. In the reaction mixture, a conversion from (E)-alpha-OHTAM to (Z)-alpha-OHTAM through the carbocation intermediate was also detected. These results show that sulfation of alpha-OHTAM catalyzed by STa results in the formation of TAM-DNA adducts.


Optical currents in vector fields.
We present the results of computer simulation of the spatial distribution of the Poynting vector and illustrate motion of microparticles and nanoparticles in spatially inhomogeneously polarized fields. The influence of phase relations and the degree of mutual coherence of superimposing waves on the characteristics of the microparticle's motion has been analyzed. For the first time, we have shown experimentally the possibility of diagnostics of optical currents in liquids caused by polarization characteristics of an optical field alone, using test metallic particles of nanoscale.


Conformational changes at cytoplasmic intersubunit interactions control Kir channel gating.
The defining structural feature of inward-rectifier potassium (Kir) channels is the unique Kir cytoplasmic domain. Recently we showed that salt bridges located at the cytoplasmic domain subunit interfaces (CD-Is) of eukaryotic Kir channels control channel gating via stability of a novel inactivated closed state. The cytoplasmic domains of prokaryotic and eukaryotic Kir channels show similar conformational rearrangements to the common gating ligand, phosphatidylinositol bisphosphate (PIP2), although these exhibit opposite coupling to opening and closing transitions. In Kir2.1, mutation of one of these CD-I salt bridge residues (R204A) reduces apparent PIP2 sensitivity of channel activity, and here we show that Ala or Cys substitutions of the functionally equivalent residue (Arg-165) in the prokaryotic Kir channel KirBac1.1 also significantly decrease sensitivity of the channel to PIP2 (by 5-30-fold). To further understand the structural basis of CD-I control of Kir channel gating, we examined the effect of the R165A mutation on PIP2-induced changes in channel function and conformation. Single-channel analyses indicated that the R165A mutation disrupts the characteristic long interburst closed state of reconstituted KirBac1.1 in giant liposomes, resulting in a higher open probability due to more frequent opening bursts. Intramolecular FRET measurements indicate that, relative to wild-type channels, the R165A mutation results in splaying of the cytoplasmic domains away from the central axis and that PIP2 essentially induces opposite motions of the major β-sheet in this channel mutant. We conclude that the removal of stabilizing CD-I salt bridges results in a collapsed state of the Kir domain.


Helminthic pseudotumours of the bowel: thirty-four cases of helminthoma.
Human infestation with nematode worms of the superfamily Strongyloidea has been recorded from time to time to give rise to serious surgical complications. Worms of the genus Oesophagostomum are most frequently responsible. These are common parasites of ruminants, monkeys, and apes in which their histotropic phase is confined to the bowel wall and sometimes results in multiple inflammatory nodules. Man is an accidental host and it seems an abnormal one. The worm fails to return to the bowel lumen, migrates further and persists in the tissues. The commonest manifestation is a solitary tumour-like inflammatory mass or abscess (;helminthoma') in the ileocaecal region. The ileum, transverse and sigmoid colons are affected less commonly and the lesions are occasionally multiple. Patients may also present with abscesses of the abdominal wall. The clinical diagnosis is difficult, even at laparotomy. Carcinoma, appendicitis, ileocaecal tuberculosis are frequently simulated and unnecessary radical surgery is often the result, particularly in expatriate Europeans. In this communication 34 cases from Uganda are reviewed with emphasis on histopathology as responsibility for the correct diagnosis is likely to fall on pathologists. Three characteristic appearances are described and related to phases in the natural history of the disease. Current knowledge on parasitology is reviewed. The disease affects Africans as well as Europeans and it is anticipated that cases will be seen in those returning from the tropics.


Dovitinib preferentially targets endothelial cells rather than cancer cells for the inhibition of hepatocellular carcinoma growth and metastasis.
Dovitinib is a receptor tyrosine kinase (RTK) inhibitor targeting vascular endothelial growth factor receptors, fibroblast growth factor receptors and platelet-derived growth factor receptor β. Dovitinib is currently in clinical trials for the treatment of hepatocellular carcinoma (HCC). In this study, we used five HCC cell lines and five endothelial cell lines to validate molecular and cellular targets of dovitinib. Tumor growth and pulmonary metastasis were significantly suppressed in an orthotopic HCC model. Immunoblotting revealed that among known dovitinib targets, only PDGFR-β was expressed in two HCC cell lines, while four of five endothelial lines expressed PDGFR-β, FGFR-1, and VEGFR-2. Dovitinib inhibited endothelial cell proliferation and motility at 0.04 μmol/L, a pharmacologically relevant concentration; it was unable to inhibit the proliferation or motility of HCC cells at the same concentration. Immunohistochemical analyses showed that dovitinib significantly decreased the microvessel density of xenograft tumors, inhibiting proliferation and inducing apoptosis in HCC cells. Our findings indicate that dovitinib inhibits HCC growth and metastasis preferentially through an antiangiogenic mechanism, not through direct targeting of HCC cells.


Acquired inguinal hernia in the horse: a review of 27 cases.
Twenty-seven cases of acquired inguinal hernia in the horse were reviewed. The history, clinical signs, treatment, and complications were summarized. Mortality was 25.9%. Only 13 or the 27 cases were diagnosed prior to admission. With early diagnosis and treatment, mortality should approach zero. In 24 of these cases, the hernia was strangulated. In each case, the signs were colic, a firm slightly enlarged testicle in the scrotum, and small intestine displacement through the inguinal ring, as determined via rectal palpation. In was concluded that palpation of the scrotum and inguinal rings should be performed on every stallion with colic. The importance of early diagnosis in decreasing mortality was illustrated by the comparison of nonsurvivors to survivors.


Discrete GPCR-triggered endocytic modes enable β-arrestins to flexibly regulate cell signaling.
β-Arrestins are master regulators of cellular signaling that operate by desensitizing ligand-activated G-protein-coupled receptors (GPCRs) at the plasma membrane and promoting their subsequent endocytosis. The endocytic activity of β-arrestins is ligand dependent, triggered by GPCR binding, and increasingly recognized to have a multitude of downstream signaling and trafficking consequences that are specifically programmed by the bound GPCR. However, only one biochemical 'mode' for GPCR-mediated triggering of the endocytic activity is presently known - displacement of the β-arrestin C-terminus (CT) to expose clathrin-coated pit-binding determinants that are masked in the inactive state. Here, we revise this view by uncovering a second mode of GPCR-triggered endocytic activity that is independent of the β-arrestin CT and, instead, requires the cytosolic base of the β-arrestin C-lobe (CLB). We further show each of the discrete endocytic modes is triggered in a receptor-specific manner, with GPCRs that bind β-arrestin transiently ('class A') primarily triggering the CLB-dependent mode and GPCRs that bind more stably ('class B') triggering both the CT and CLB-dependent modes in combination. Moreover, we show that different modes have opposing effects on the net signaling output of receptors - with the CLB-dependent mode promoting rapid signal desensitization and the CT-dependent mode enabling prolonged signaling. Together, these results fundamentally revise understanding of how β-arrestins operate as efficient endocytic adaptors while facilitating diversity and flexibility in the control of cell signaling.


Current and future perspectives on fulvestrant.
Fulvestrant, an estrogen receptor antagonist with no known agonist effects, is effective and well tolerated in the treatment of hormone-sensitive breast cancer after antiestrogen failure in postmenopausal women. Numerous phase II and III clinical trials of fulvestrant that are designed to build on its efficacy in breast cancer and explore its value in other tumors are ongoing or in the final planning stage. Favorable safety, dose-response, and pharmacokinetic data led to the initiation of clinical trials to evaluate loading and higher doses with the aim of building on the well-defined efficacy of fulvestrant. Recently reported results of phase II trials by the North Central Cancer Treatment Group and the Swiss Group for Clinical Cancer Research support the clinical activity of fulvestrant after recurrence or progression on a nonsteroidal aromatase inhibitor, and 2 international phase III trials are ongoing in this setting. As a first-line treatment for metastatic disease, fulvestrant is currently being evaluated in combination with anastrozole versus anastrozole alone in 2 phase III trials, 1 by the Southwest Oncology Group and the other being conducted mainly in Scandinavia. Preclinical data have also led to randomized phase II trials of fulvestrant in combination with the epidermal growth factor receptor tyrosine kinase inhibitor, gefitinib, and the HER2/neu-targeted antibody trastuzumab. Additional phase II and III trials are currently evaluating fulvestrant plus tipifarnib, Theratope vaccine, or the dual kinase inhibitor, GW572016. Although fulvestrant is undergoing robust clinical development as a treatment for breast cancer, investigation of this agent in other types of solid tumors has only just begun.


Is genetically modified crop the answer for the next green revolution?
Post-green revolution advances made in biotechnology paved the way of cultivating the high-yielding, stress and disease resistant genetically modified (GM) varieties of wheat, rice, maize cotton and several other crops. The recent rapid commercialization of the genetically modified crops in Asia, Americas and Australia indicates the potentiality of this new technology. GM crops give higher yields and are rich in nutritional values containing vitamins and minerals and can thus can help to alleviate hunger and malnutrition of the growing population in the under developed and developing countries. It could also be possible to develop more biotic and abiotic stress resistant genotypes in these crops where it was difficult to develop due to the unavailability of genes of resistance in the crossing germplasms. However, further research and investigations are needed to popularize the cultivation of these crops in different parts of the world. This review provides an insight of the impact of GM crops on contemporary agriculture across the past few decades, traces its' history across time, highlights new achievements and breakthroughs and discusses the future implication of this powerful technology in the coming few decades.


Protective effect of Chuquiraga spinosa Lessing associated with simvastatin on N-Nitroso-N-methylurea (NMU)-induced prostate cancer in rats.
Chuquiraga spinosa Lessing (ChS) has shown protective effect on N-Nitroso-N-methylurea (NMU)-induced prostate cancer in rats. Currently, statins are being studied for their pro-apoptotic and antimetastatic effects. The main objective of this research was to determine the protective effect associated with the oral administration of simvastatin and ethanolic extract of the aerial parts of ChS in the prevention of prostate cancer. Fifty-six albino male rats were randomized into seven groups: I) negative control: physiological serum: 2 mL/kg; II) TCN: testosterone 100 mg/kg + cyproterone 50 mg/kg + NMU 50 mg/kg; III) TCN + S40 (simvastatin 40 mg/kg); IV) TCN + ChS250 (ChS 250 mg/kg); V) TCN + ChS50 (ChS 50 mg/kg) + S40; VI) TCN + ChS250 (ChS 250 mg/kg) + S40; and VII) TCN + ChS500 (ChS 500 mg/kg) + S40. The antioxidant activity was tested by using (2,2-diphenyl-1-picrylhydrazyl) (DPPH) assay. Hematology, toxicological biochemical parameters, prostate-specific antigen (PSA), histology and prostate size were evaluated as main indicators of protective effect. Triglyceride values were decreased in the groups receiving ChS, being significant (P=0.02) in IV and VII group compared to cancer-inducing group (TCN). In groups that received ChS, PSA levels (P=0.71) were significant compared with TCN group. The VII group had the lowest prostate volume by sonography. The TCN group showed multiple foci of high-grade prostatic intraepithelial neoplasia (HG-PIN) with the presence of cells in mitosis; whilst, groups V and VI had few areas of HG-PIN. In experimental conditions, the ethanolic extract of C. spinosa in association with simvastatin showed a protective effect on prostate cancer through hypolipidemic and antioxidant activity.


Stromal signatures in endometrioid endometrial carcinomas.
The pattern of myometrial invasion in endometrioid endometrial carcinomas varies considerably; ie, from widely scattered glands and cell nests, often associated with a fibromyxoid stromal reaction (desmoplasia) and/or a lymphocytic infiltrate, to invasive glands with little or no stromal response. Recently, two distinct stromal signatures derived from a macrophage response (colony-stimulating factor 1, CSF1) and a fibroblastic response (desmoid-type fibromatosis, DTF) were identified in breast carcinomas and correlated with clinicopathologic features including outcome. In this study, we explored whether these stromal signatures also apply to endometrioid carcinomas and how their expression patterns correlated with morphologic changes. We studied the stromal signatures both by immunohistochemistry and in situ hybridization in 98 primary endometrioid carcinomas with (87 cases) and without (11 cases) myometrial invasion as well as in the corresponding regional lymph nodes metatases of 9 myoinvasive tumors. Desmoplasia correlated positively with the DTF expression signature. Likewise, mononuclear infiltrates were found in the stroma of tumors expressing CSF1. Twenty-four out of eighty-seven (27%) myoinvasive endometrioid carcinomas were positive for the macrophage signature and thirteen out of eighty-seven (15%) expressed the fibroblast signature. Eleven additional cases were positive for both DTF and CSF1 signatures (11/87; 13%). However, over half of the cases (39/87; 45%) and the majority of the non-myoinvasive tumors (8/11; 73%) failed to express any of the two stromal signatures. The macrophage response (CSF1) was associated with higher tumor grade, lymphovascular invasion, and PIK3CA mutations (P<0.05). There was a concordance in the expression of the CSF1 signature in the primary tumors and their corresponding lymph node metastases. This study is the first characterization of stromal signatures in endometrioid carcinomas. Our findings shed new light on the relationship between genetically different endometrioid carcinomas and various stromal responses. Preservation of the CSF1 macrophage stromal response in the metastases leds support to targeting the CSF1 pathway in endometrioid endometrial carcinomas.


Clinical, social, and policy factors in COVID-19 cases and deaths: methodological considerations for feature selection and modeling in county-level analyses.
There is a need to evaluate how the choice of time interval contributes to the lack of consistency of SDoH variables that appear as important to COVID-19 disease burden within an analysis for both case counts and death counts. This study identified SDoH variables associated with U.S county-level COVID-19 cumulative case and death incidence for six different periods: the first 30, 60, 90, 120, 150, and 180 days since each county had COVID-19 one case per 10,000 residents. The set of SDoH variables were in the following domains: resource deprivation, access to care/health resources, population characteristics, traveling behavior, vulnerable populations, and health status. A generalized variance inflation factor (GVIF) analysis was used to identify variables with high multicollinearity. For each dependent variable, a separate model was built for each of the time periods. We used a mixed-effect generalized linear modeling of counts normalized per 100,000 population using negative binomial regression. We performed a Kolmogorov-Smirnov goodness of fit test, an outlier test, and a dispersion test for each model. Sensitivity analysis included altering the county start date to the day each county reached 10 COVID-19 cases per 10,000. Ninety-seven percent (3059/3140) of the counties were represented in the final analysis. Six features proved important for both the main and sensitivity analysis: adults-with-college-degree, days-sheltering-in-place-at-start, prior-seven-day-median-time-home, percent-black, percent-foreign-born, over-65-years-of-age, black-white-segregation, and days-since-pandemic-start. These variables belonged to the following categories: COVID-19 related, vulnerable populations, and population characteristics. Our diagnostic results show that across our outcomes, the models of the shorter time periods (30 days, 60 days, and 900 days) have a better fit. Our findings demonstrate that the set of SDoH features that are significant for COVID-19 outcomes varies based on the time from the start date of the pandemic and when COVID-19 was present in a county. These results could assist researchers with variable selection and inform decision makers when creating public health policy.


Relationship between ADD1 Gly460Trp gene polymorphism and essential hypertension in Madeira Island.
Essential hypertension (EH) is a complex disease in which physiological, environmental, and genetic factors are involved in its genesis. The genetic variant of the alpha-adducin gene (ADD1) has been described as a risk factor for EH, but with controversial results.The objective of this study was to evaluate the association of ADD1 (Gly460Trp) gene polymorphism with the EH risk in a population from Madeira Island.A case-control study with 1614 individuals of Caucasian origin was performed, including 817 individuals with EH and 797 controls. Cases and controls were matched for sex and age, by frequency-matching method. All participants collected blood for biochemical and genotypic analysis for the Gly460Trp polymorphism. We further investigated which variables were independently associated to EH, and, consequently, analyzed their interactions.In our study, we found a significant association between the ADD1 gene polymorphism and EH (odds ratio 2.484, P = .01). This association remained statistically significant after the multivariate analysis (odds ratio 2.548, P = .02).The ADD1 Gly460Trp gene polymorphism is significantly and independently associated with EH risk in our population. The knowledge of genetic polymorphisms associated with EH is of paramount importance because it leads to a better understanding of the etiology and pathophysiology of this pathology.


Selective suppression of retroviral gp70-anti-gp70 immune complex formation by prostaglandin E1 in murine systemic lupus erythematosus.
The effect of pharmacologic quantities of prostaglandin E1 (PGE) was investigated in three strains of mice (NZB X NZW, MRL/1, and BXSB) that spontaneously develop lupus-like glomerulonephritis. PGE-treatment prolonged survival and retarded the glomerular deposition of immune complex (IC) and the development of glomerulonephritis in NZB X NZW and MRL/1 mice, but did not similarly protect BXSB mice. Changes in the responsive strains correlated well with reduced amounts of circulating gp70 complexed with anti-gp70 antibodies compared with untreated controls, although total concentrations of gp70 (free and complexed) detectable in sera were similar in both groups of mice. The results strongly suggest that: (a) PGE selectively suppressed the immune response to retroviral gp70, (b) PGe had little effect on the quantity or quality of anti-DNA antibodies but did reduce the deposition of anti-DNA containing IC in the kidneys, and (c) gp70 IC appear to play an important role in the pathogenesis of glomerulonephritis in murine systemic lupus erythematosus.


The kinetics of electrical activity of beta cells in response to a "square wave" stimulation with glucose or glibenclamide.
The effect of a "square wave" stimulation with glucose or glibenclamide on the electrical activity of beta-cells has been studied with microelectrode techniques in isolated perifused mouse islets. While glucose evoked a burst activity with periodic oscillations of the membrane potential between two levels, glibenclamide produced a constant depolarization with a continuous spike activity. In both cases the time course of activity was biphasic and resembled closely the corresponding patterns of insulin release. The effect of glibenclamide on the membrane was irreversible. The results provide further evidence for a direct correlation between insulin release and electrical activity. Furthermore, it is suggested that the membrane potential is, at least in part, involved in the mechanism of biphasic insulin release.


Nicotine stimulates DNA synthesis and proliferation in vascular endothelial cells in vitro.
Nicotine is a major component of cigarette smoke and has been postulated to play an important role in atherogenesis and malignancy. Endothelial cell growth may be regulated by nicotine, yet operative mechanisms at the endothelial level are poorly understood. We studied the effects of nicotine (10(-14)-10(-4) M) on endothelial DNA synthesis, DNA repair, proliferation, and cytotoxicity by using cultures of bovine pulmonary artery endothelial cells. Assays were performed on cells incubated with nicotine in the presence and absence of hydroxyurea (an inhibitor of scheduled DNA synthesis), serum, human platelet-poor plasma, and platelet-derived growth factor and endothelial cell growth factor (PDGF and PDECGF, respectively). Nicotine significantly stimulated endothelial cell DNA synthesis and proliferation at concentrations lower than those obtained in blood after smoking (<10(-8) M). The stimulatory effects of nicotine were enhanced by serum (0.5%) and PDECGF and were blocked by the nicotinic-receptor antagonist hexamethonium. The response to nicotine was bimodal because cytotoxicity was observed at higher concentrations (>10(-6) M). This study has implications for understanding cellular mechanisms of nicotine action. The results may be important in tumor angiogenesis, atherogenesis, and vascular dysfunction in smokers.


Molecular identification of Salp15, a key salivary gland protein in the transmission of lyme disease spirochetes, from Ixodes persulcatus and Ixodes pacificus (Acari: Ixodidae).
Salp15 is a multifunctional protein, vital to the tick in its need to obtain vertebrate host blood without stimulating a host inflammatory and immune response. The Salpl5 protein from both Ixodes scapularis Say and Ixodes ricinus (L.), the principal vectors of the Lyme disease spirochete in eastern North America and Europe, respectively, have been well characterized and found to bind the murine CD4 receptor, DC-SIGN, and the OspC protein of Borrelia burgdorferi. In the current study, we characterized the full salp15 gene in Ixodes pacificus Cooley & Kohls and Ixodes persulcatus Schulze, the principal vectors of Lyme disease spirochetes in western North America and Asia, respectively. In comparing the Salp15 protein of all four principal vector ticks of public health importance for the transmission of Lyme disease spirochetes, we find the 53 C-terminal amino acids to have a high degree of similarity. There are at least three clades in the tree of Salp15 and its homologues, probably representing a multigene family.


Applying genetic algorithm optimization to a folded geometry acousto-optic modulated spatial pulse shaper.
A folded geometry acousto-optic modulator spatial pulse shaper has been designed for shaping individual pulses from a high power amplified laser. The design preserves the capability of computer programmable amplitude and phase modulation of femtosecond laser pulses. An additional application of genetic algorithm optimization approach for compressing a stretched pulse is also demonstrated for such a pulse shaper. Spectrally and temporally resolved optical gating technique is used to characterize the shaped pulses.


Factors influencing quality of life in breast cancer survivors.
Longitudinal data from 195 breast cancer survivors were used to identify factors affecting the level and rate of change in quality of life after completion of treatment. Women were interviewed up to four times at approximately yearly intervals using Kaplan and Bush's Quality of Well Being instrument (QWB). Random coefficient regression analysis was used to model QWB as a function of time since diagnosis and personal characteristics. QWB scores decreased over time and the rate of decline increased with age (p = 0.032). This was similar to declines in women with benign breast biopsies, but overall QWB levels were lower in women with breast cancer. Having a spouse tended to slow the rate of decline in breast cancer survivors (p = 0.004). The presence of comorbidity was associated with significantly lower QWB levels (p = 0.037) but did not affect the rate of change over time. Education, family history of breast cancer, cancer stage and treatment modalities were not significantly related to QWB levels or rates of change. Breast cancer survivors experience a reduction in quality of life that persists for years after treatment and is similar in magnitude to that associated with other health problems.


Metabolic compensation of fitness costs associated with overexpression of the multidrug efflux pump MexEF-OprN in Pseudomonas aeruginosa.
The acquisition of antibiotic resistance has been associated with a possible nonspecific, metabolic burden that is reflected in decreased fitness among resistant bacteria. We have recently demonstrated that overexpression of the MexEF-OprN multidrug efflux pump does not produce a metabolic burden when measured by classical competitions tests but rather leads to a number of changes in the organism's physiology. One of these changes is the untimely activation of the nitrate respiratory chain under aerobic conditions. MexEF-OprN is a proton/substrate antiporter. Overexpression of this element should result in a constant influx of protons, which may lead to cytoplasmic acidification. Acidification was not observed in aerobiosis, a situation in which the MexEF-overproducing mutant increases oxygen consumption. This enhanced oxygen uptake serves to eliminate intracellular proton accumulation, preventing the cytoplasmic acidification that was observed exclusively under anaerobic conditions, a situation in which the fitness of the MexEF-OprN-overproducing mutant decreases. Finally, we determined that the early activation of the nitrate respiratory chain under aerobic conditions plays a role in preventing a deleterious effect associated with the overexpression of MexEF-OprN. Our results show that metabolic rewiring may assist in overcoming the potential fitness cost associated with the acquisition of antibiotic resistance. Furthermore, the capability to metabolically compensate for this effect is habitat dependent, as demonstrated by our results under anaerobic conditions. The development of drugs that prevent metabolic compensation of fitness costs may help to reduce the persistence and dissemination of antibiotic resistance.


Activation of pp60c-src protein kinase activity in human colon carcinoma.
The tyrosine-specific protein kinase activity of pp60c-src molecules obtained from human colon carcinoma tissues and tumor-derived cell lines was found to be elevated over that from normal colon tissues or cultures of normal colon mucosal cells. The elevated pp60c-src protein kinase activity in tumor tissues and in cultured colon carcinoma cells does not appear to result solely from an increase in the abundance of the c-src-encoded protein, suggesting that the specific activity of the pp60c-src tyrosine phosphotransferase is enhanced. These results raise the possibility that activation of the pp60c-src protein kinase may contribute to the genesis of human colon tumors.


Impact of multiplanar review of three-dimensional echocardiographic data on management of congenital heart disease.
In patients with congenital cardiac malformations, accurate diagnosis is critical in diagnosis and management. The multiplanar review mode (MPR) allows the operator to cut three-dimensional (3D) echocardiographic data sets in infinite planes, and to review the moving image in three simultaneous orthogonal planes. We sought to describe the clinical utility of MPR of 3D echocardiography for analysis of congenitally malformed hearts. Cross-sectional and 3D MPR echocardiography was performed in 300 patients with congenitally malformed hearts. Analysis in multiplanar mode was possible in all patients. New, clinically important information, which altered management or changed the principal diagnosis, was obtained in 32 (11%) cases. This determined suitability for biventricular repair in 11 patients, clarified the morphology of atrioventricular valves in 7, helped in assessment of aortic, mitral, or prosthetic valvar disease in 13, and identified a vascular ring in the other patient. 3D MPR is feasible in the setting of the congenitally malformed heart, permitting focused and in-depth analysis. This substantially improves the understanding of functional morphology, above the information derived from cross-sectional echocardiography. We recommend the use of the 3D format with MPR for patients with complex congenital cardiac disease.


Glutathione-dependent redox status of frataxin-deficient cells in a yeast model of Friedreich's ataxia.
Friedreich's ataxia is a neurodegenerative disease caused by reduced expression of the mitochondrial protein frataxin. The main phenotypic features of frataxin-deficient human and yeast cells include iron accumulation in mitochondria, iron-sulphur cluster defects and high sensitivity to oxidative stress. Glutathione is a major protective agent against oxidative damage and glutathione-related systems participate in maintaining the cellular thiol/disulfide status and the reduced environment of the cell. Here, we present the first detailed biochemical study of the glutathione-dependent redox status of wild-type and frataxin-deficient cells in a yeast model of the disease. There were five times less total glutathione (GSH+GSSG) in frataxin-deficient cells, imbalanced GSH/GSSG pools and higher glutathione peroxidase activity. The pentose phosphate pathway was stimulated in frataxin-deficient cells, glucose-6-phosphate dehydrogenase activity was three times higher than in wild-type cells and this was coupled to a defect in the NADPH/NADP(+) pool. Moreover, analysis of gene expression confirms the adaptative response of mutant cells to stress conditions and we bring evidence for a strong relation between the glutathione-dependent redox status of the cells and iron homeostasis. Dynamic studies show that intracellular glutathione levels reflect an adaptation of cells to iron stress conditions, and allow to distinguish constitutive stress observed in frataxin-deficient cells from the acute response of wild-type cells. In conclusion, our findings provide evidence for an impairment of glutathione homeostasis in a yeast model of Friedreich's ataxia and identify glutathione as a valuable indicator of the redox status of frataxin-deficient cells.


A rational design strategy of the novel topoisomerase II inhibitors for the synthesis of the 4-O-(2-pyrazinecarboxylic)-4'-demethylepipodophyllotoxin with antitumor activity by diminishing the relaxation reaction of topoisomerase II-DNA decatenation.
A rational design strategy of the novel podophyllum topoisomerase II (Topo II) inhibitors for the synthesis of the esterification and amidation substituted 4'-demethylepipodophyllotoxin (DMEP) derivates was developed in order to discover the potential antitumor prodrug. Firstly, according to the structure-activity relationship, drug combination principle and bioisosterism, the -COO- and the -NH- bond substituents at the 4 position of cycloparaffin would be a great modification direction to improve antitumor activity of 4'-demethylepipodophyllotoxin (DMEP). Secondly, from the prodrug principle view, the esterification and amidation at the C-4 position of DMEP would be two useful structure modifications for improve solubility. Thirdly, from the activity pocket in Topo II-DNA cleavage complex point of view, a series of heterocyclic with pharmacological activity were chosen as module for improving antitumor activity by binding with Topo II. Finally, nine novel esterification and amidation DMEP derivates were designed and synthesized for the potential Topo II inhibitors with the superior biological activity. All the novel compounds exhibited promising in vitro antitumor activity, especially 4-O-(2-pyrazinecarboxylic)-4'-demethylepipodophyllotoxin (compound 1). The antitumor activity of compound 1 against tumor cell line HeLa (i.e., the IC50 value of 0.60 ± 0.20 μM), A549 (i.e., the IC50 value of 3.83 ± 0.08 μM), HepG2 (i.e., the IC50 value of 1.21 ± 0.05 μM), and BGC-823 (i.e., the IC50 value of 4.15 ± 1.13 μM) was significantly improved by 66, 16, 12, and 6 times than that of the clinically important podophyllum anticancer drug etoposide (i.e., the IC50 values of 15.32 ± 0.10, 59.38 ± 0.77, 67.25 ± 7.05, and 30.74 ± 5.13 μM), respectively. Compound 1 could arrest HeLa cell cycle G2/M and induce apoptosis by strongly diminishing the relaxation reaction of Topo II-DNA decatenation. The correctness of rational drug design was strictly demonstrated by the bioactivity test.


Pediatric community acquired urinary tract infections due to extended-spectrum beta-lactamase versus non-extended-spectrum beta-lactamase producing bacteria.
This study aimed to evaluate the demographics, clinical characteristics, risk factors, and antibiotic resistance of pediatric community-acquired urinary tract infections (UTIs) caused by extended-spectrum beta-lactamase (ESBL)-producing and non-ESBL-producing uropathogens. This retrospective study was conducted at a tertiary care hospital in Saudi Arabia, among children aged between 0 and 14 years, with a culture-proven diagnosis of community-acquired UTI between February 2019 and September 2021. Patients were divided into two groups based on whether or not their UTI was caused by ESBL-producing bacteria. A total of 383 patients with community-acquired UTI were evaluated. Escherichia coli was detected in 72.6% of cultures. Extended-spectrum beta-lactamase-producing organisms were responsible for 35.7% of UTI episodes. Of these 69% and 31% were caused by E. coli and Klebsiella pneumoniae, respectively. There were no significant differences between the two groups with regard to clinical presentation or urine analysis. The resistance rates in the ESBL-producing group were 39.4% for amoxicillin/clavulanic acid, 65.7% for ciprofloxacin, 72.3% for co-trimoxazole, 32.8% for nitrofurantoin, 21.2% for gentamicin, and 0.7% for amikacin and carbapenems. In the non-ESBL-producing group, it was 22.4% for amoxicillin/clavulanic acid, 22.4% for ciprofloxacin, 38.2% for co-trimoxazole, 23.6% for nitrofurantoin, 6.1% for gentamicin, and zero for amikacin and carbapenems. The presence of renal abnormalities (p = 0.014) and male gender (p = 0.026) were determined to be independent risk factors for ESBL UTIs. Recognizing risk factors and antibiotic resistance for ESBL-producing bacteria may aid in tailoring an antibiotic regimen for pediatric patients at high risk of ESBL-UTIs.


Supra-biological factors in the assessment of males seeking penile prostheses.
The technique of installing penile prostheses for treatment of erectile dysfunction in the male is becoming increasingly popular, particularly, but by no means exclusively in the cases of impotence in which biological factors are clearly implicated. The literature to date fails to take into account suprabiological factors such as the nature of the couple's sexual relationship and more particularly the sexual myths that shape their sexual behaviour. The author evaluated eight couples in which the male was seeking a penile prostheses and in all eight couples a variety of sexual myths had contributed to major sexual problems for the couple, even before the onset of the erectile dysfunction. A plea is made for discarding the false dichotomy or "organic" versus "functional", in favour of a list of etiological factors including the biological ones as well as suprabiological ones. Also, the installation of penile implants should only be considered after a full investigation of the couple's sexuality by a competent sex therapist and only when other forms of couple/sex therapy have failed.


Effect of cryo-protective agents on the growth and ultrastructure of Saccharomyces cerevisiae.
When suspended in a glucose medium supplemented with varying amounts of DMSO, ethylene glycol, glycerol, lactose and sodium chloride at its optimum temperature of 30 degrees C, a diploid strain of Saccharomyces cerevisiae gave a mean generation time inversely proportional to the increased concentrations of these anti-freeze compounds. No growth was attained at 0 degrees C and -5 degrees C, and mixtures of the compounds did not support any growth even at 30 degrees C. At 0 degrees C and -5 degrees C with increasing percentages of the individual compounds, and at 30 degrees C, 0 degrees C and -5 degrees C, with mixtures of the compounds, a significant decrease in survival was obtained. As compared to cells grown in the absence of antifreeze compounds, electron microscopy of the cells which grew at 30 degrees C in the presence of antifreeze compounds showed thicker cell walls, highly convoluted plasmalemma, vacuoles filled with electron-dense fibrous material, spherosomes, poorly developed mitochondria and many vesicles.


Influence of particle size distribution, organic carbon, pH and chlorides on washing of mercury contaminated soil.
Feasibility of soil washing to remediate Hg contaminated soil was studied. Dry sieving was performed to evaluate Hg distribution in soil particle size fractions. The influence of dissolved organic matter and chlorides on Hg dissolution was assessed by batch leaching tests. Mercury mobilization in the pH range of 3-11 was studied by pH-static titration. Results showed infeasibility of physical separation via dry sieving, as the least contaminated fraction exceeded the Swedish generic guideline value for Hg in soils. Soluble Hg did not correlate with dissolved organic carbon in the water leachate. The highest Hg dissolution was achieved at pH 5 and 11, reaching up to 0.3% of the total Hg. The pH adjustment was therefore not sufficient for the Hg removal to acceptable levels. Chlorides did not facilitate Hg mobilization under acidic pH either. Mercury was firmly bound in the studied soil thus soil washing might be insufficient method to treat the studied soil.


Curcumin, a potent anti-tumor reagent, is a novel histone deacetylase inhibitor regulating B-NHL cell line Raji proliferation.
To investigate curcumin (diferuloylmethane) induced apoptosis and its molecular mechanism of action in B-NHL cell line Raji cells. Raji cells were cultured in RPMI-1640 medium and treated with curcumin in different concentrations. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) assay was used to detect growth inhibition and Hoechst 33258 staining was used to detect apoptosis. Immunocytochemistry and Western blot were used to detect the expressions of histone deacetylase 1, 3, and 8 (HDAC1, HDAC3, and HDAC8) and acetylated histone H4 (Ac-histone H4) protein. Curcumin inhibited the proliferation of B-NHL cell line Raji cells with a 36-h IC50 value of 24.1+/-2.0 micromol/L. Hoechst 33258 staining showed that curcumin could induce Raji cell apoptosis. The expression levels of HDAC1, HDAC3, and HDAC8 proteins were downregulated following curcumin treatment in Raji cells, whereas Ac-histone H4 protein expression was upregulated after treatment with curcumin. Curcumin, as a new member of the histone deacetylase inhibitors, can inhibit the expression of class I HDACs (HDAC1, HDAC3, and HDAC8), and can increase the expression of Ac-histone H4 in Raji cells. Curcumin plays an important role in regulating B-NHL cell line Raji cell proliferation and apoptosis.


Generation of induced pluripotent stem cells from human Tenon's capsule fibroblasts.
This study aimed to develop a feasible and efficient method for generating embryonic stem cell (ESC)-like induced pluripotent stem (iPS) cells from human Tenon's capsule fibroblasts (HTFs) through the expression of a defined set of transcription factors, which will have significant application value for ophthalmic personalized regenerative medicine. HTFs were harvested from fresh samples, and reprogramming was induced by the exogenous expression of the four classic transcription factors, OCT-3/4, SOX-2, KLF-4, and C-MYC. The HTF-derived iPS (TiPS) cells were analyzed with phase contrast microscopy, real-time PCR, immunofluorescence, FACS analysis, alkaline phosphatase activity analysis, and a teratoma formation assay. Human ESC colonies were used as the positive control. The resulting HTF-derived iPS cell colonies were indistinguishable from human ESC colonies regarding morphology, gene expression levels, pluripotent gene expression, alkaline phosphatase activity, and the ability to generate all three embryonic germ layers. This study presents a simple, efficient, practical procedure for generating patient-tailored iPS cells from HTFs. These cells will serve as a valuable and preferred candidate donor cell population for ophthalmological regenerative medicine.


Impact of nuclear accidents on marine biota.
The accident at the Fukushima Daiichi nuclear power plant, precipitated by the earthquake and subsequent tsunami that struck the northeastern coast of Japan in March 2011, has raised concerns about the potential impact to marine biota posed by the release of radioactive water and radionuclide particles into the environment. The Fukushima accident is the only major nuclear accident that has resulted in the direct discharge of radioactive materials into a coastal environment. This article briefly summarizes what is currently understood about the effects of radioactive wastewaters and radionuclides to marine life.


Quantified relations between exposure to tobacco smoking and bladder cancer risk: a meta-analysis of 89 observational studies.
Smoking is a major risk factor for bladder cancer (BC). This meta-analysis updates previous reviews on smoking characteristics and BC risk, and provides a more quantitative estimation of the dose-response relationship between smoking characteristics and BC risk. In total, 89 studies comprising data from 57 145 BC cases were included and summary odds ratios (SORs) were calculated. Dose-response meta-analyses modelled relationships between smoking intensity, duration, pack-years and cessation and BC risk. Sources of heterogeneity were explored and sensitivity analyses were conducted to test the robustness of findings. Current smokers (SOR = 3.14, 95% CI = 2.53-3.75) and former smokers(SOR = 1.83, 95% CI = 1.52-2.14) had an increased risk of BC compared with never smokers. Age at first exposure was negatively associated with BC risk. BC risk increased gradually by smoking duration and a risk plateau at smoking 15 cigarettes a day and 50 pack-years was observed. Smoking cessation is most beneficial from 20 years before diagnosis. The population-attributable risk of BC for smokers has decreased from 50% to 43% in men and from 35% to 26% in women from Europe since estimated in 2000. Results were homogeneous between sources of heterogeneity, except for lower risk estimates found in studies of Asian populations. Active smokers are at an increased risk of BC. Dose-response meta-analyses showed a BC risk plateau for smoking intensity and indicate that even after long-term smoking cessation, an elevated risk of bladder cancer remains.


Comparison of manual and automatic sampling for monitoring ochratoxin A in barley grain.
Automatic and manual sampling for ochratoxin A (OTA) in barley grain was compared under industrial conditions considering sampling uncertainty as well as practical and technical aspects. Ten tonnes of barley inoculated with Penicillium verrucosum were incubated until the OTA concentration reached approximately 15 µg kg(-1) and sampled with manual and automatic sampling. A nested experimental design and ANOVA was used to estimate variance components from sampling, sample reduction, sample preparation and analysis. Manual sampling resulted in a high sampling uncertainty and OTA concentrations in aggregate samples ranged from 2 to 80 µg kg(-1). When aggregate samples were formed by automatic sampling the uncertainty arising from nugget effects and spatial distribution was practically eliminated. Results from this study show that an automatic sampler mounted after a mixer or conveyer can provide representative samples of OTA from a moving stream of barley. Automatic sampling might present a practical and economical alternative to manual sampling for feed mill operators when monitoring low levels of mycotoxins in grain or other commodities. Despite careful precautions, sample preparation and analysis resulted in a relative uncertainty of ±40% (p = 0.95), which was attributed to the sub-sampling following the two grinding steps. Size fractionation of the coarsely ground barley showed that 40% of the total amount of OTA was present in a small fraction of fine particles with a strong tendency to aggregate or stick to equipment and containers. Thus, in order to take advantage of the automatic sampling, it is crucial to apply an appropriate sub-sampling to prevent segregation of particles which may affect the OTA measurements.


Protective effect of quercetin on liver damage induced by chronic toluene exposure in rats.
The aim of this study was to evaluate the possible protective effects of quercetin (QE) on liver injury caused by chronic toluene exposure in rats. The rats were randomly allotted into one of three experimental groups: control, toluene treated and toluene treated with QE; each group contained 10 animals. The control group received 1 ml physiologic serum, and toluene treatment was performed by inhalation of 3000 ppm toluene, for 8 h/day and 6 days/week for 12 weeks. The rats in the QE-treated groups were given QE (15 mg/kg, intraperitoneal) once a day for 12 weeks starting just after toluene exposure. Tissue samples were obtained for histopathological investigation. To date, no histopathological changes in the liver of rats after chronic toluene exposure by QE treatment have been reported. Light microscopic evaluation of liver tissue samples of toluene-exposed rats revealed enlarged sinusoids filled with blood. In addition, some of the hepatocytes showed loss of cytoplasm, and some had a hyperchromatic nucleus. QE treatment attenuated alterations in liver histology. The alpha smooth muscle actin, transforming growth factor beta-positive cells and the activity of terminal deoxynucleotidyl transferase dUTP nick end labeling in the toluene-treated group were observed to be reduced with QE treatment. The data indicate that QE attenuates toluene-induced liver injury, but further investigation is still absolutely necessary.


All primary care trainees are not the same: the role of economic factors and career choice.
Historically, the specialties classified as "primary care" have been pediatrics, internal medicine, and family medicine. Often, primary care disciplines are aggregated in workforce or career-preference studies, and any differences among them are not assessed or reported. However, such aggregation is likely unwarranted and may actually lead to false or misguided policy direction in the name of "primary care disciplines" when, in fact, there may be substantive differences among these specialties. We examine here the data available to assess whether the physicians who make up 2 of the primary care specialties are necessarily the same with regard to their perspectives on the economic aspects of medicine in general, or their own personal economic preferences regarding career choice in particular. Recent research has demonstrated that there are substantive differences among internal medicine trainees and pediatric trainees. As such, we must be cautious when using data from 1 primary care specialty to suggest structural and/or policy changes regarding the economic structure of training for the others.


Xolik: finding cross-linked peptides with maximum paired scores in linear time.
Cross-linking technique coupled with mass spectrometry (MS) is widely used in the analysis of protein structures and protein-protein interactions. In order to identify cross-linked peptides from MS data, we need to consider all pairwise combinations of peptides, which is computationally prohibitive when the sequence database is large. To alleviate this problem, some heuristic screening strategies are used to reduce the number of peptide pairs during the identification. However, heuristic screening strategies may miss some true cross-linked peptides. We directly tackle the combination challenge without using any screening strategies. With the data structure of double-ended queue, the proposed algorithm reduces the quadratic time complexity of exhaustive searching down to the linear time complexity. We implement the algorithm in a tool named Xolik. The running time of Xolik is validated using databases with different numbers of proteins. Experiments using synthetic and empirical datasets show that Xolik outperforms existing tools in terms of running time and statistical power. Source code and binaries of Xolik are freely available at http://bioinformatics.ust.hk/Xolik.html. Supplementary data are available at Bioinformatics online.


Blockade of Adrenal Medulla-Derived Epinephrine Potentiates Bee Venom-Induced Antinociception in the Mouse Formalin Test: Involvement of Peripheral β -Adrenoceptors.
The injection of diluted bee venom (DBV) into an acupoint has been used traditionally in eastern medicine to treat a variety of inflammatory chronic pain conditions. We have previously shown that DBV had a potent antinociceptive efficacy in several rodent pain models. However, the peripheral mechanisms underlying DBV-induced antinociception remain unclear. The present study was designed to investigate the role of peripheral epinephrine on the DBV-induced antinociceptive effect in the mouse formalin assay. Adrenalectomy significantly enhanced the antinociceptive effect of DBV during the late phase of the formalin test, while chemical sympathectomy had no effect. Intraperitoneal injection of epinephrine blocked this adrenalectomy-induced enhancement of the DBV-induced antinociceptive effect. Moreover, injection of a phenylethanolamine N-methyltransferase (PNMT) inhibitor enhanced the DBV-induced antinociceptive effect. Administration of nonselective β -adrenergic antagonists also significantly potentiated this DBV-induced antinociception, in a manner similar to adrenalectomy. These results demonstrate that the antinociceptive effect of DBV treatment can be significantly enhanced by modulation of adrenal medulla-derived epinephrine and this effect is mediated by peripheral β -adrenoceptors. Thus, DBV acupoint stimulation in combination with inhibition of peripheral β -adrenoceptors could be a potentially novel strategy for the management of inflammatory pain.


Inhibition of hepatitis B Dane particle DNA polymerase activity by pyrophosphate analogs.
A DNA polymerase is associated with the core of the so-called Dane particles. The probability that this is the hepatitis B viral DNA polymerase offers the possibility of preventing hepatitis B multiplication by selective inhibition of this enzyme. We have previously reported that trisodium phosphonoformate (PFA) inhibits Dane particle DNA polymerase. Fifteen compounds with structural similarity to PFA and pyrophosphate have now been tested for inhibition of hepatitis B virus DNA polymerase in an attempt to define the structural requirement for the inhibition. Active structures have two acid groups at close proximity of which at least one is a phosphono group. Phosphonoformate and hypophosphare were the two most active inhibitors. The Ki value for PFA was 7.2 microM when dTTP was used as variable substrate, and the mechanism of inhibition was non-competitive. Phosphonoformate caused rapid shut-off of the polymerase reaction, indicating that it might inhibit elongation. The efficient inhibition of hepatitis B virus DNA polymerase by PFA and its low toxicity suggest that it could be used to inhibit hepatitis B virus multiplication in vivo.


Substrate adhesion of rat hepatocytes: on the mechanism of attachment to fibronectin.
We examined the mechanisms of cell attachment to fibronectin-coated substrates. Inhibition of cell attachment was obtained by species-specific antifibronectin antibodies, which presumably recognize a distinct antigenic structure in the protein located at, or in the immediate vicinity of, the cell-binding site. The inhibiting antibodies could be adsorbed on a column of Sepharose substituted with plasma fibronectin. The initial phase of cell attachment was also inhibited by addition of soluble fibronectin to the incubation medium in a reaction that exhibited specificity and concentration dependence. These data suggest that cell-binding sites are available in an active form on the surface of soluble fibronectin. However, the inhibitory effect of fibronectin was greatly enhanced by adding the protein together with heparin, heparan sulfate, collagen, or a fibronectin-binding collagen peptide (CB-7), which is consistent with an "activation" of fibronectin on binding to these matrix components. A similar activation of fibronectin was obtained by cleaving the protein with trypsin. We discuss these findings in relation to conformational rearrangements in the fibronectin molecule. Data is presented supporting a mechanism of cell attachment to fibronectin involving multiple weak interactions between cellular receptors and substrate molecules, although some steps in the attachment process appear to disobey the requirements for such a mechanism.


Coherent Spin Dynamics of Electrons in CsPbBr3 Perovskite Nanocrystals at Room Temperature.
Coherent spin dynamics of charge carriers in CsPbBr3 perovskite nanocrystals are studied in a temperature range of 4-300 K and in magnetic fields of up to 500 mT using time-resolved pump-probe Faraday rotation and differential transmission techniques. We detect electron spin Larmor precession in the entire temperature range. At temperatures below 50 K, hole spin precession is also observed. The temperature dependences of spin-related parameters, such as Landè g-factor and spin dephasing time are measured and analyzed. The electron g-factor increases with growing temperature, which can not be described by the temperature-induced band gap renormalization. We find that photocharging of the nanocrystals with either electrons or holes depends on the sample cooling regime, namely the cooling rate and illumination conditions. The type of the charge carrier provided by the photocharging can be identified via the carrier spin Larmor precession.


Ferromagnetic Anomalous Hall Effect in Cr-Doped Bi2Se3 Thin Films via Surface-State Engineering.
The anomalous Hall effect (AHE) is a nonlinear Hall effect appearing in magnetic conductors, boosted by internal magnetism beyond what is expected from the ordinary Hall effect. With the recent discovery of the quantized version of the AHE, the quantum anomalous Hall effect (QAHE), in Cr- or V-doped topological insulator (TI) (Sb,Bi)2Te3 thin films, the AHE in magnetic TIs has been attracting significant interest. However, one of the puzzles in this system has been that while Cr- or V-doped (Sb,Bi)2Te3 and V-doped Bi2Se3 exhibit AHE, Cr-doped Bi2Se3 has failed to exhibit even ferromagnetic AHE, the expected predecessor to the QAHE, though it is the first material predicted to exhibit the QAHE. Here, we have successfully implemented ferromagnetic AHE in Cr-doped Bi2Se3 thin films by utilizing a surface state engineering scheme. Surprisingly, the observed ferromagnetic AHE in the Cr-doped Bi2Se3 thin films exhibited only a positive slope regardless of the carrier type. We show that this sign problem can be explained by the intrinsic Berry curvature of the system as calculated from a tight-binding model combined with a first-principles method.


Nonsimultaneous stimulus presentations and their role in listener naming.
Previous studies on naming have presented the object and its name simultaneously during both training and testing, and thus the training component may establish a transformation of function directly between the object and the name. Successful tests for listener naming may thus not require the emergence of a novel (entailed) transformation of function. The current study aimed to control for this possibility by presenting the object and the name sequentially and nonsimultaneously. Eight typically developing toddlers participated in the current study. During name training, objects and names were presented nonsimultaneously, and all participants failed to emit listener-naming responses during the first test session. Subsequently, 4 participants received multiple exemplar training, which led to improvements in listener naming for all 4; and speaker naming for only 1 participant. As a control condition, the remaining 4 participants were tested repeatedly, without multiple exemplar training, and did not show any consistent improvements in their listener or speaker performances. Multiple exemplar training thus appeared to be effective in establishing generalized listener responses, which involved generating entailed transformation of functions. The strategy of using nonsimultaneous stimulus presentations could allow for greater precision in identifying the behavioral processes involved in listener-naming.


Women and men who have served in Afghanistan/Iraq: coming home.
The experience of war changes people - some will acknowledge that the changes are positive and some will feel the opposite or a combination, but that it changes a person cannot be disputed. For those who return, the experience of reintegration to civilian life or as a respite before redeployment can present numerous challenges. The research presented in this article reports the findings on interviews with over 800 service members who had returned from either Afghanistan or Iraq. The Post Deployment Reintegration Scale was used to refine the areas that respondents identified as positive or negative in their reintegration experience. Implications for practice with returning service members are noted.


Proteasome inhibition in skeletal muscle cells unmasks metabolic derangements in type 2 diabetes.
Two-dimensional difference gel electrophoresis (2-D DIGE)-based proteome analysis has revealed intrinsic insulin resistance in myotubes derived from type 2 diabetic patients. Using 2-D DIGE-based proteome analysis, we identified a subset of insulin-resistant proteins involved in protein turnover in skeletal muscle of type 2 diabetic patients, suggesting aberrant regulation of the protein homeostasis maintenance system underlying metabolic disease. We then validated the role of the ubiquitin-proteasome system (UPS) in myotubes to investigate whether impaired proteasome function may lead to metabolic arrest or insulin resistance. Myotubes derived from muscle biopsies obtained from people with normal glucose tolerance (NGT) or type 2 diabetes were exposed to the proteasome inhibitor bortezomib (BZ; Velcade) without or with insulin. BZ exposure increased protein carbonylation and lactate production yet impaired protein synthesis and UPS function in myotubes from type 2 diabetic patients, marking the existence of an insulin-resistant signature that was retained in cultured myotubes. In conclusion, BZ treatment further exacerbates insulin resistance and unmasks intrinsic features of metabolic disease in myotubes derived from type 2 diabetic patients. Our results highlight the existence of a confounding inherent abnormality in cellular protein dynamics in metabolic disease, which is uncovered through concurrent inhibition of the proteasome system.


Intracellular substrates for the primer-unblocking reaction by human immunodeficiency virus type 1 reverse transcriptase: detection and quantitation in extracts from quiescent- and activated-lymphocyte subpopulations.
Treatment of human immunodeficiency virus type 1 (HIV-1)-infected patients with 3'-azido-3'-deoxythymidine (AZT) selects for mutant forms of viral reverse transcriptase (RT) with increased ability to remove chain-terminating nucleotides from blocked DNA chains. We tested various cell extracts for the presence of endogenous acceptor substrates for this reaction. Cell extracts incubated with HIV-1 RT and [(32)P]ddAMP-terminated DNA primer/template gave rise to (32)P-labeled adenosine 2',3'-dideoxyadenosine 5',5'''-P(1),P(4)-tetraphosphate (Ap(4)ddA), ddATP, Gp(4)ddA, and Ap(3)ddA, corresponding to the transfer of [(32)P]ddAMP to ATP, PP(i), GTP, and ADP, respectively. Incubation with [(32)P]AZT monophosphate (AZTMP)-terminated primer/template gave rise to the analogous (32)P-labeled AZT derivatives. Based on the rates of formation of the specific excision products, ATP and PP(i) levels were determined: ATP was present at 1.3 to 2.2 mM in H9 cells, macrophages, and unstimulated CD4(+) or CD8(+) T cells, while PP(i) was present at 7 to 15 microM. Under these conditions, the ATP-dependent reaction predominated, and excision by the AZT-resistant mutant RT was more efficient than wild type RT. Activated CD4(+) or CD8(+) T cells contained 1.4 to 2.7 mM ATP and 55 to 79 microM PP(i). These cellular PP(i) concentrations are lower than previously reported; nonetheless, the PP(i)-dependent reaction predominated in extracts from activated T cells, and excision by mutant and wild-type RT occurred with similar efficiency. While PP(i)-dependent excision may contribute to AZT resistance in vivo, it is likely that selection of AZT-resistant mutants occurs primarily in an environment where the ATP-dependent reaction predominates.


Conservative management following closed reduction of traumatic anterior dislocation of the shoulder.
Acute anterior dislocation, which is the most common type of shoulder dislocation, usually results from an injury. Subsequently, the shoulder is less stable and is more susceptible to re-dislocation, especially in active young adults. This is an update of a Cochrane review first published in 2006. To assess the effects (benefits and harms) of conservative interventions after closed reduction of traumatic anterior dislocation of the shoulder. These might include immobilisation, rehabilitative interventions or both. We searched the Cochrane Bone, Joint and Muscle Trauma Group Specialised Register (September 2013), the Cochrane Central Register of Controlled Trials (CENTRAL) (2013, Issue 8), MEDLINE (1946 to September 2013), EMBASE (1980 to Week 38, 2013), CINAHL (1982 to September 2013), PEDro (1929 to November 2012), OTseeker (inception to November 2012) and trial registries. We also searched conference proceedings and reference lists of included studies. Randomised or quasi-randomised controlled trials comparing various conservative interventions versus control (no or sham treatment) or other conservative interventions applied after closed reduction of traumatic anterior dislocation of the shoulder. All review authors independently selected trials, assessed risk of bias and extracted data. Study authors were contacted for additional information. Results of comparable groups of trials were pooled. We included three randomised trials and one quasi-randomised trial, which involved 470 participants (371 male) with primary traumatic anterior dislocation of the shoulder reduced by various closed methods. Three studies evaluated mixed populations; in the fourth study, all participants were male and 80% were soldiers. All trials were at some risk of bias but to a differing extent. One was at high risk in all domains of the risk of bias tool, and one was at unclear or high risk in all domains; the other two trials were deemed to have predominantly low risk across all domains. Overall, reflecting both the risk of bias and the imprecision of findings, we judged the quality of evidence to be "very low" for all outcomes, meaning that we are very uncertain about the estimates of effect.The four trials evaluated the same comparison - immobilisation in external rotation versus internal rotation - and each of our three primary outcomes (re-dislocation, patient-reported outcome measures (PROMs) for shoulder instability and resumption of activities) was reported by one or more of the trials, with two-year or longer follow-up. Pooling was possible for "re-dislocation" (three trials) and for aspects of "resumption of sport/activities at pre-injury level" (two trials).There was no evidence to show a difference between the two groups in re-dislocation at two-year or longer follow-up (risk ratio (RR) 1.06 favouring internal rotation, 95% confidence interval (CI) 0.73 to 1.54; P value 0.77; 252 participants; three trials). In a low-risk population, with an illustrative baseline risk of 247 re-dislocations per 1000, these data equate to 15 more (95% CI 67 fewer to 133 more) re-dislocations per 1000 after immobilisation in external rotation. In a medium-risk population, with an illustrative baseline risk of 436 re-dislocations per 1000, the data equate to 26 more (95% CI 118 fewer to 235 more) re-dislocations after immobilisation in external rotation.Nor was evidence found to show a difference between the two groups in return to pre-injury levels of activity at two-year or longer follow-up (RR 1.25 favouring external rotation, 95% CI 0.71 to 2.2; P value 0.43; 278 participants; two trials). In a low-risk population, with an illustrative baseline risk of 204 participants per 1000 returning to pre-injury levels of activity, this equates to 41 more (95% CI 59 fewer to 245 more) participants per 1000 resuming activity after immobilisation in external rotation. In a high-risk population, with an illustrative baseline risk of 605 participants per 1000 returning to pre-injury levels of activity, this equates to 161 more (95% CI 76 fewer to 395 more) participants per 1000 resuming activity after immobilisation in external rotation.One trial reported that the difference between the two groups in Western Ontario Shoulder Instability Index scores, analysed using non-parametric statistics, was "not significant (P = 0.32)". Of our secondary outcomes, pooling was possible for "any instability" (two trials) and for important adverse events (three events, two trials). However, adverse event data were collected only in an ad hoc way, and it is unclear whether identification and reporting of such events was comprehensive. No report addressed participant satisfaction or health-related quality of life outcome measures.There was no evidence confirming a difference between the two positions of immobilisation in any of the primary or secondary outcomes; for each outcome, the confidence intervals were wide, covering the possibility of substantial benefit for each intervention. Numerous conservative strategies may be adopted after closed reduction of a traumatic anterior dislocation of the shoulder, and many warrant investigation. However, our review reveals that evidence from randomised controlled trials is only available for a single approach: immobilisation in external rotation versus immobilisation in the traditional position of internal rotation. Moreover, this evidence is insufficient to demonstrate whether immobilisation in external rotation confers any benefit over immobilisation in internal rotation.We identified six unpublished trials and two ongoing trials that compare immobilisation in external versus internal rotation. Given this, the main priority for research on this question consists of the publication of completed trials, and the completion and publication of ongoing trials. Meanwhile, increased attention to other interventions is required. Sufficiently powered, good quality, well reported randomised controlled trials with long-term surveillance should be conducted to examine the optimum duration of immobilisation, whether immobilisation is necessary at all (in older age groups particularly), which rehabilitative interventions work best and the acceptability to participants of different care strategies.


A paired comparison of tuberculin skin test results in health care workers using 5 TU and 10 TU tuberculin.
Historically, 10 TU has been employed in Australia and the United Kingdom to perform the tuberculin skin test (TST). However, this makes it difficult to compare the rates of TST positivity with other countries such as the USA who use 5 TU. To assess the impact of the dose of tuberculin on the TST a comparison was made of TST responses in health care workers given a TST with both 5 and 10 TU. Two TSTs were performed simultaneously in each health care worker using 5 and 10 TU. Each dose was randomly assigned in a blinded manner to the right or left forearm and read at 48-72 hours by a single nurse who was blinded to the assignment of the 5 and 10 TU doses. A total of 128 health care workers were enrolled, 119 (93%) of whom had a past history of BCG vaccination. The overall mean difference in paired reaction sizes for the two doses was 1.5 mm with 95% limits of agreement of -3.6 to 6.5 mm. A slightly larger TST reading was seen with 10 TU than with 5 TU. The mean difference of 1.5 mm between the two doses should be considered when comparing rates of TST positivity between countries who use different doses of tuberculin to perform the tuberculin skin test.


Participant and Caregiver Perspectives on Clinical Research During Covid-19 Pandemic.
The COVID-19 pandemic has massively disrupted essential clinical research. Many regulatory organizations have rightfully advocated to temporarily halt enrollment and curtail all face-to-face interactions. Views and opinions of patients and their caregivers are seldom considered while making such decisions. The objective was to study older participants' and their caregivers' perspectives to participate in ongoing clinical research during the COVID-19 pandemic. Cross-sectional. VISN-16/Geriatric Research, Education and Clinical Center (GRECC), Department of Veterans Affairs. Older participants and their caregivers (N = 51) enrolled in ongoing clinical research studies. Questions about perceptions of safety to attend research visit, the level of panic among the general public, and medical center's preparedness in handling the pandemic. Other questions identified the source of pandemic information and the preference of a phone or in-person visit. Mean age was 69.3 (±9.4) years, 53% were male, 39% were caregivers, and 65% were Caucasian. Majority (78%) of the participants felt safe/very safe attending the scheduled research appointment; 63% felt that the extra screening made them feel safe/very safe; 82% felt that the medical center was prepared/very prepared for the pandemic. Participants split evenly on their preference for phone versus in-person visits. Family members and television news media were the commonly used sources of pandemic information irrespective of their education. Perceptions were influenced by gender and source of information, not by age or education. Females perceived higher level of panic compared to males (P = .02). Those relying on news media felt safer compared to those that relied on family members (P = .008). Even though informants felt that the medical center was prepared to handle the pandemic, only half the participants preferred the in-person visit. Pandemic information was obtained from family members or the television news media. Knowing patients' perspectives may help researchers be better prepared for future pandemics. J Am Geriatr Soc 68:E14-E18, 2020.


Adventitious Virus Detection in Cells by High-Throughput Sequencing of Newly Synthesized RNAs: Unambiguous Differentiation of Cell Infection from Carryover of Viral Nucleic Acids.
The use of high-throughput sequencing (HTS) to identify viruses in biologicals differs from current molecular approaches, since its use enables an unbiased approach to detection without the need to design specific primers to preamplify target sequences. Its broad range of detection and analytical sensitivity make it an important tool to ensure that biologicals are free from adventitious viruses. Similar to other molecular methods, however, identification of viral sequences in cells by HTS does not prove viral infection, since this could reflect carryover of inert viral sequences from reagents or other sources or the presence of transcriptionally inactive cellular sequences. Due to the broad range of detection associated with HTS, the above can potentially be perceived as a drawback for the testing of pharmaceutical biological products using this method. In order to avoid the identification of inert viral sequences, we present a methodology based on metabolic RNA labeling and sequencing, which enables the specific identification of newly synthesized viral RNAs in infected cells, resulting in the ability to unambiguously distinguish active infection by DNA or RNA viruses from inert nucleic acids. In the present study, we report the ability to differentiate Vero cells acutely infected by a single-stranded positive-sense RNA virus (tick-borne encephalitis virus) from cells which have been in contact with nonreplicating virus particles. Additionally, we also found a laboratory contamination by the squirrel monkey retrovirus of our Vero cell line, which was derived from an Old World (African green) monkey, a type of contamination which until now has been identified only in cells derived from primates from the New World.IMPORTANCE The use of high-throughput sequencing (HTS) to identify viral contamination of biological products is extremely sensitive and provides a broad range of detection. Nevertheless, viral sequences identified can also be inert. Examples include contamination resulting from reagents or the presence of inactivated viruses in animal-derived components of the cell culture medium. We therefore developed a method that relies on the sequencing of newly synthesized RNAs, an unequivocal sign of the presence of a transcriptionally active virus. This improvement in the specificity of viral testing increases the acceptability of HTS as a standard test for cells used in manufacturing biologicals and in biotherapies.


Hole Transfer Processes in meta- and para-Conjugated Mixed Valence Compounds: Unforeseen Effects of Bridge Substituents and Solvent Dynamics.
To address the question whether donor substituents can be utilized to accelerate the hole transfer (HT) between redox sites attached in para- or in meta-positions to a central benzene bridge, we investigated three series of mixed valence compounds based on triarylamine redox centers that are connected to a benzene bridge via alkyne spacers at para- and meta-positions. The electron density at the bridge was tuned by substituents with different electron donating or accepting character. By analyzing optical spectra and by DFT computations we show that the HT properties are independent of bridge substituents for one of the meta-series, while donor substituents can strongly decrease the intrinsic barrier in the case of the para-series. In stark contrast, temperature-dependent ESR measurements demonstrate a dramatic increase of both the apparent barrier and the rate of HT for strong donor substituents in the para-cases. This is caused by an unprecedented substituent-dependent change of the HT mechanism from that described by transition state theory to a regime controlled by solvent dynamics. For solvents with slow longitudinal relaxation (PhNO2, oDCB), this adds an additional contribution to the intrinsic barrier via the dielectric relaxation process. Attaching the donor substituents to the bridge at positions where the molecular orbital coefficients are large accelerates the HT rate for meta-conjugated compounds just as for the para-series. This effect demonstrates that the para-meta paradigm no longer holds if appropriate substituents and substitution patterns are chosen, thereby considerably broadening the applicability of meta-topologies for optoelectronic applications.


Metabolism of arsenic by sheep chronically exposed to arsenosugars as a normal part of their diet. 1. Quantitative intake, uptake, and excretion.
Information on the effects of long-term organoarsenical consumption by mammals is limited despite the fact that foodstuffs, especially seafood, often contain organoarsenicals at very high concentrations. Here we evaluate the intake, uptake, and excretion (urine and feces) of arsenic by sheep that live on North Ronaldsay in the Orkney Islands and naturally consume large amounts of arsenosugars through their major food source-seaweed. The sheep eat a broad variety of seaweed species, and arsenic concentrations were determined in all the species observed eaten by the sheep (5.7-74.0 mg kg(-1) dry mass). Because of preference and availability, they feed mostly on the seaweed species found to contain the highest arsenic concentrations: Laminaria digitata and Laminaria hyperborea (74 +/- 4 mg kg(-1) dry mass). To quantify the arsenic intake by the sheep, a feeding experiment reflecting natural conditions as close as possible was set up. In the feeding trial, the average daily intake of arsenic by 12 ewes was 35 +/- 6 mg (97% of water-extractable arsenic was present as arsenosugars) gained from feeding on the two brown algae. To test the possible influence of microflora on the metabolism of arsenosugars, six of the sheep were adapted to feeding on grass for 5 months before the start of the trial (control sheep), and the remaining six sheep were kept on their normal seaweed diet (wild sheep). No significant difference in seaweed/arsenic intake and arsenic excretion was found between the two groups of sheep. The arsenic excreted in the feces represents 13 +/- 10% (n = 12) of the total consumed, and on the assumption of that, the average urinary excretion is estimated to 86%.The main arsenic metabolite excreted in urine was dimethylarsinic acid (DMA(V)) (60 +/- 22%) and minor amounts of dimethylarsinoylethanol (DMAE), methylarsonic acid (MA(V)),tetramethylarsonium ion (TMA+), and arsenate (As(V)) together with seven unknown arsenic compounds were also excreted. The urinary arsenic excretion pattern showed a lag period (>4 h) before significant quantities appeared in the urine, an excretion rate that peaked between 4 and 28 h after seaweed intake and a relatively slow half-life (17 h) after end of intake.