Miracle-dz/UniFAR-FTUs huggingface dataset:
A novel laboratory technique demonstrating the influences of RHD zygosity and the RhCcEe phenotype on erythrocyte D antigen expression

D antigen is the most immunogenic and clinically relevant antigen within the complex Rh blood group system. Variability of D antigen expression was first described decades ago but has rarely been investigated quantitatively, particularly in the context of RHD zygosity along with RhCcEe serological phenotype. With IRB approval, 107 deidentified blood samples were analyzed. Rh phenotypes were determined serologically by saline technique using monoclonal antibodies against D, C, c, E, and e antigens. RHD zygosity was determined using both PCR‐restriction fragment length polymorphisms and quantitative real‐time PCR techniques. A novel and robust method was developed for quantitation of erythrocyte D antigen sites using calibrated microspheres and flow cytometry, allowing correlation of D antigen density with RHD zygosity and expression of Rh CcEe antigens. Subjects homozygous for RHD expressed nearly twice the number of D antigen sites compared with RHD hemizygotes (33,560 ± 8,222 for DD versus 17,720 ± 4,471 for Dd, P < 0.0001). Expression of c or E antigens was associated with significantly increased erythrocyte D antigen expression, whereas presence of C or e antigens reduced expression. These data and this novel quantitation method will be important for future studies investigating the clinical relevance of D antigen variability. Am. J. Hematol. , 2012. © 2011 Wiley Periodicals, Inc.
Effects of Chymase on the Macular Region in Monkeys and Porcine Müller Cells: Probable Involvement of Chymase in the Onset of Idiopathic Macular Holes

Objectives: To investigate chymase involvement in idiopathic macular hole onset, the effects of chymase on monkey eyes and cultured Müller cells were investigated. Methods: Immunohistochemistry using antinestin and antiglial fibrillary acidic protein antibodies was performed in a normal monkey eye. After chymase was injected into the monkey vitreous, histological changes in the retina were evaluated using the TdT-mediated dUTP nick-end labeling (TUNEL) assay. Expression of c-kit, a stem cell factor receptor, and nestin was examined in porcine Müller cells cultured with basic fibroblast growth factor. The effects of chymase on proliferation and TUNEL staining in Müller cells were also examined. Results: The number of nestin and glial fibrillary acidic protein-positive cells was higher in the macula than in other regions. Thickening of the posterior hyaloid membrane and some apoptotic cells were found in the macula of chymase-treated eyes. The expression of c-kit and nestin in Müller cells was shown and enhanced when cultured with basic fibroblast growth factor. Exposure to chymase inhibited Müller cell proliferation and produced TUNEL-positive cells. Conclusions: There might be Müller cells possessing atypical properties near the macular region and chymase might cause fibrosis and apoptosis through these cells. These findings suggest that increased chymase activity may result in idiopathic macular hole onset.
Comparison of single‐shot localization methods (steam and press) for In vivo proton NMR spectroscopy

Two single‐shot localization techniques, STEAM and PRESS, are analyzed with regard to specifications for in vivo localized proton NMR. In particular, attention is paid to optimum signal intensity per unit volume, sensitivity to motion and diffusion, shortest attainable echo time, water suppression and editing possibilities. Experimental results are shown for cat brain at 4.7 T and human brain at 1.5 T. Both STEAM and PRESS are highly effective localization methods. For long echo times, PRESS is the method of choice, because it offers a factor of two gain in signal intensity. In addition, the method is less sensitive to motion and diffusion, and not susceptible to multiple‐quantum effects. STEAM offers advantages for observation of (coupled) metabolites with short T2, because (a) shorter TEs can be attained and (b) effective water suppression sequences can be implemented without penalty in echo time. Differences relating to editing possibilities and B1 dependence, possibly important in choosing a method, are discussed.
Analysis of a recombinant mouse hepatitis virus expressing a foreign gene reveals a novel aspect of coronavirus transcription

We have inserted heterologous genetic material into the nonessential gene 4 of the coronavirus mouse hepatitis virus (MHV) in order to test the applicability of targeted RNA recombination for site-directed mutagenesis of the MHV genome upstream of the nucleocapsid (N) gene and to develop further genetic tools for site-directed mutagenesis of structural genes other than N. Initially, a 19-nucleotide tag was inserted into the start of gene 4a of MHV strain A59 with the N gene deletion mutant Alb4 as the recipient virus. In further work, the entire gene for the green fluorescent protein (GFP) was inserted in place of gene 4, creating the currently largest known RNA virus. The expression of GFP was demonstrated by Western blot analysis of infected cell lysates; however, the level of GFP expression was not sufficient to allow detection of fluorescence of viral plaques. Northern blot analysis of transcripts of GFP recombinants showed the expected alteration of the pattern of the nested MHV subgenomic mRNAs. Surprisingly, though, GFP recombinants also produced an RNA species that was the same size as wild-type mRNA4. Analysis of the 5' end of this species revealed that it was actually a collection of mRNAs originating from 10 different genomic fusion sites, none possessing a canonical intergenic sequence. The finding of these aberrant mRNAs suggests that long-range RNA or the ribonucleoprotein structure of the MHV genome can sometimes be the sole determinant of the site of initiation of transcription.
Evidence-based interventions to reduce tuberculosis stigma: a systematic review.

SETTING While substantial progress is being made in tuberculosis (TB) control, the success of public health efforts is hampered by pervasive stigma. OBJECTIVE To perform a systematic literature review to assess the effectiveness of interventions aimed at reducing TB stigma in patients, health care workers, care givers and the general community. DESIGN Studies were eligible for inclusion if they evaluated interventions aimed at reducing TB stigma and were published between 1950 and 2015. We searched eight databases (PubMed, Cochrane Library, Ovid, Embase, PsycInfo, Sociological Abstracts, Cumulative Index to Nursing and Allied Health Literature, World Health Organization Latin American and Caribbean Health Sciences Literature), and complemented the searches by using the snowball strategy and by reviewing relevant grey literature. RESULTS Only seven studies were identified as providing quantitative (n = 4) or qualitative (n = 3) evidence of effectiveness in reducing TB stigma. Quality assessment of the studies was poor. Knowledge-shaping and attitude-changing interventions aimed at the public, patients and their families were effective in reducing anticipated stigma. Home visits and support groups were effective in reducing both anticipated and internalised stigma. CONCLUSION There is a dearth of reliable information on the effectiveness of TB stigma-reduction interventions. Knowledge-shaping, attitude-changing and patient-support interventions can be effective in reducing TB stigma, but more rigorous evaluations are needed.
Establishment and characterization of a continuous cell line from pupal ovaries of Japanese oak silkworm Antheraea Yamamai Guerin-Meneville

SummaryPupal ovaries of the wild oak silkworm Antheraea yamamai Guerin-Meneville were cultured in MGM-448 (Modified Grace Medium-448) medium containing 10% fetal bovine serum. After the primary culture was set up in 1988, a continuous cell line was obtained in 1991, designated as NISES-Anya-0611 (Anya-0611). The population doubling time was 54 hrs. and 19 min. at 96 passages and 88 hrs. and 29 min. at 387 passages. Spindle-shaped and spherical cells coexisted in the cell group. The cell line karyotype line was typical of lepidopteran cell lines, consisting of numerous small chromosomes. The cell line was distinguished from other lepidopteran cell lines by comparing malic enzyme, phosphoglucose isomerase, phosphoglucose mutase, and isocitric dehydrogenase isozyme patterns. The cell line was highly infected to the Antheraea yamamai nuclear polyhedrosis virus (Anya NPV). The luciferase gene of recombinant Bm NPV (BmNPVP6ETL) was able to express in the cell line, too, so that luciferase recombinant products were able to be detected in the cell body and in supernatant. The Anya NPV clone group was isolated on the cell seat using plaque purification.
Determination of acute and early life stage toxicity of fat‐plant effluent using zebrafish (Danio rerio)

The present study examines the effects of an effluent from a fat plant (FP) on zebrafish (Danio rerio) embryos and larvae using the whole effluent toxicity testing methods (WET). The method is based on acute toxicity using 96‐h larval mortality and chronic toxicity using endpoints such as the time to hatch, hatching success, deformity, growth rate, swim‐up failure, accumulative mortality, and sex ratio. On the basis of larval mortality the 96‐h LC50 (the concentration was lethal to 50% of newly hatching zebrafish larvae) was 68.9%. In chronic toxicity test, newly fertilized embryos (<5‐h old) were exposed to 1, 6, 12, 25, 50% effluent concentrations in a 24‐h static renewal system at (27 ± 0.5)°C until 15‐day posthatch. The results showed that all chronic endpoints were significantly different from the control at 50% dilution. Embryos began to show lesions on third day at higher concentrations (12, 25, 50% FP effluent concentrations). Treatment group of 25% dilution showed delayed time to hatch. Morphological abnormalities were observed in newly hatched larvae at 25 and 50% FP effluent concentrations. At 25% dilution, sex ratio of larvae was alternated and there was feminization phenomenon. On the basis of the study, the FP effluent tested here may cause increasing embryotoxicity in the zebrafish embryos. We conclude that the test using zebrafish is feasible to evaluate both acute and chronic toxicities of industrial effluents. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2008.
Fusobacterium nucleatum and Human Beta-Defensins Modulate the Release of Antimicrobial Chemokine CCL20/Macrophage Inflammatory Protein 3α

ABSTRACT Cells of the innate immune system regulate immune responses through the production of antimicrobial peptides, chemokines, and cytokines, including human beta-defensins (hBDs) and CCL20. In this study, we examined the kinetics of primary human oral epithelial cell (HOEC) production of CCL20 and hBDs in response to Fusobacterium nucleatum, a commensal bacterium of the oral cavity, which we previously showed promotes HOEC induction of hBDs. HOECs secrete maximal levels of CCL20 at 6 h, following stimulation by F. nucleatum cell wall (FnCW). The kinetics of CCL20 release is distinct from that of hBD-2 and -3, which peaks after 24 h and 48 h of FnCW stimulation, respectively. FnCW-induced release of CCL20 by HOECs requires both transcriptional and translational activation. Release of CCL20 by HOECs is inhibited by brefeldin A, suggesting that it is secreted through a vesicle transport pathway. Other epithelium-derived agents that FnCW induces, such as hBD-2, hBD-3, tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β), are also able to release CCL20. By focusing on mitogen-activated protein kinases, we show that both extracellular signal-regulated kinase 1/2 and p38, but not JNK, are required for hBD-, TNF-α-, and IL-1β-induced secretion of CCL20 by HOECs. The ability of FnCW and its induced hBDs to produce proinflammatory cytokines and CCL20 suggests the broad role of F. nucleatum and human antimicrobial peptides in primary immune responses elicited by oral epithelium.
Psychotherapies for Panic Disorder: A Tale of Two Sites.

OBJECTIVE To compare cognitive-behavioral therapy (CBT), panic-focused psychodynamic psychotherapy (PFPP), and applied relaxation training (ART) for primary DSM-IV panic disorder with and without agoraphobia in a 2-site randomized controlled trial. METHOD 201 patients were stratified for site and DSM-IV agoraphobia and depression and were randomized to CBT, PFPP, or ART (19-24 sessions) over 12 weeks in a 2:2:1 ratio at Weill Cornell Medical College (New York, New York) and University of Pennsylvania ("Penn"; Philadelphia, Pennsylvania). Any medication was held constant. RESULTS Attrition rates were ART, 41%; CBT, 25%; and PFPP, 22%. The most symptomatic patients were more likely to drop out of ART than CBT or PFPP (P = .013). Outcome analyses revealed site-by-treatment interactions in speed of Panic Disorder Severity Scale (PDSS) change over time (P = .013). At Cornell, no differences emerged on improvement on the primary outcome, estimated speed of change over time on the PDSS; at Penn, ART (P = .025) and CBT (P = .009) showed greater improvement at treatment termination than PFPP. A site-by-treatment interaction (P = .016) for a priori-defined response (40% PDSS reduction) showed significant differences at Cornell: ART 30%, CBT 65%, PFPP 71% (P = .007), but not at Penn: ART 63%, CBT 60%, PFPP 48% (P = .37). Penn patients were more symptomatic, differed demographically from Cornell patients, had a 7.2-fold greater likelihood of taking medication, and had a 28-fold greater likelihood of taking benzodiazepines. However, these differences did not explain site-by-treatment interactions. CONCLUSIONS All treatments substantially improved panic disorder with or without agoraphobia, but patients, particularly the most severely ill, found ART less acceptable. CBT showed the most consistent performance across sites; however, the results for PFPP showed the promise of psychodynamic psychotherapy for this disorder. TRIAL REGISTRATION ClinicalTrials.gov identifier: NCT00353470.
PTEN methylation is associated with advanced stage and microsatellite instability in endometrial carcinoma

Loss of heterozygosity and mutations in the PTEN (MMAC1) tumor suppressor gene are frequent in endometrial carcinoma. Promoter hypermethylation has recently been identified as an alternative mechanism of tumor suppressor gene inactivation in cancer, but its importance in the PTEN gene in endometrial carcinoma is unknown. The purpose of our study was to assess the frequency of promoter methylation of the PTEN gene and to determine its correlation with clinicopathologic variables in a prospective and population‐based series of endometrial carcinomas with complete follow‐up. Presence of PTEN promoter methylation was seen in 26 of 138 patients (19%). Methylation was significantly associated with metastatic disease (p = 0.01) and a microsatellite unstable phenotype (p = 0.006). In conclusion, we find that PTEN promoter methylation is relatively frequent in endometrial carcinoma. Its association with metastatic disease and microsatellite instability implicates its importance in the development of this tumor type. Int. J. Cancer 91:22–26, 2001. © 2001 Wiley‐Liss, Inc.
The Influence of Recency of Use on fMRI Response During Spatial Working Memory in Adolescent Marijuana Users

Abstract Some neurocognitive recovery occurs within a month of abstinence from heavy marijuana use, yet functional magnetic resonance imaging (fMRI) has revealed altered activation among recent and abstinent adult users. We compared fMRl response during a spatial working memory (SWM) task between adolescent marijuana users with brief and sustained durations of abstinence. Participants were 13 recent users (two to seven days abstinent), 13 abstinent users (27 to 60 days abstinent), and 18 nonusing controls, all ages 15 to 18. Groups were similar on demographics, had no psychiatric or medical disorders, and user groups were similar on substance histories. Teens performed a two-back SWM task during fMRI. Recent users showed greater fMRI response in medial and left superior prefrontal cortices, as well as bilateral insula. Abstinent users had increased response in the right precentral gyrus (clusters ≥ 1328 μ1, p < .05). Results suggest that adolescents who recently used marijuana show increased brain activity in regions associated with working memory updating and inhibition. This study preliminarily suggests that (1) recent marijuana use may disrupt neural connections associated with SWM and result in compensatory brain response, and (2) sustained abstinence from marijuana may be associated with improvements in SWM response among adolescents.
Measuring relationship strength in online social networks based on users' activities and profile information

Using online social networks as a means of interchanging information and ideas is developing daily so that internet users spend too much time in social network websites. This fact has turned social networks into very rich sources of information for analyzing users' activities and relationships. In recent years, most research in analysis of social networks has dealt with the friendship network. Since the friendship network is binary, the analysis thereof is of no difference between two users who are very close friends or farther acquaintances. Therefore, the result of the analysis would be not real in many cases. In this paper, we tried to measure relationship strength based on users' activities and profile information. For this purpose, a Facebook application was developed so as to collect users' information. Then based on the users' answers to the questionnaire, we measured their relationship strength at four levels. After outlier removal and normalization of reminder data, some of the features were removed based on the Information Gain criterion. Then considering the collected data, two models of Decision Tree and multilayer perceptron were used so as to measure the users' relationship strength. Having applied 10-fold Cross Validation, the validity of the learned model was evaluated. Based on our experiments, multilayer perceptron could predict the users' relationship strength with 72% accuracy and the accuracy of decision tree with ID3 learning algorithm is a bit lower. Finally In two-class case, we compared MLP with SVM and found that MLP achieved 87% of classification accuracy. Based on accuracy, recall and precision we concluded that MLP can classify relationship strength better.
P-glycoprotein inhibition of drug resistant cell lines by nanoparticles

Abstract Several pharmaceutical excipients are known for their ability to interact with cell membrane lipids and reverse the phenomenon of multidrug resistance (MDR) in cancer. Interestingly, many excipients act as stabilizers and are key ingredients in a variety of nano-formulations. In this study, representatives of ionic and non-ionic excipients were used as surface active agents in nanoparticle (NP) formulations to utilize their MDR reversing potential. In-vitro assays were performed to elucidate particle–cell interaction and accumulation of P-glycoprotein (P-gp) substrates-rhodamine-123 and calcein AM, in highly drug resistant glioma cell lines. Chemosensitization achieved using NPs and their equivalent dose of free excipients was assessed with the co-administered anti-cancer drug doxorubicin. Among the excipients used, non-ionic surfactant, Cremophor® EL, and cationic surfactant, cetyltrimethylammonuium bromide (CTAB), demonstrated highest P-gp modulatory activity in both free solution form (up to 7-fold lower IC50) and as a formulation (up to 4.7-fold lower IC50) as compared to doxorubicin treatment alone. Solutol® HS15 and Tween® 80 exhibited considerable chemosensitization as free solution but not when incorporated into a formulation. Sodium dodecyl sulphate (SDS)-based nanocarriers resulted in slightly improved cytotoxicity. Overall, the results highlight and envisage the usage of excipient in nano-formulations in a bid to improve chemosensitization of drug resistant cancer cells towards anti-cancer drugs.
Automated Neutral Loss and Data Dependent Energy Resolved “Pseudo MS3” for the Targeted Identification, Characterization and Quantitative Analysis of Methionine-Containing Peptides

A strategy involving the fixed-charge sulfonium ion derivatization, stable isotope labeling, capillary high-performance liquid chromatography and automated data dependent neutral loss scan mode tandem mass spectrometry (MS/MS) and “pseudo multiple mass spectrometry (MS3)” product ion scans in a triple quadrupole mass spectrometer has been developed for the “targeted” gas-phase identification, characterization and quantitative analysis of low abundance methionine-containing peptides present within complex protein digests. Selective gas-phase “enrichment” and identification is performed via neutral loss scan mode MS/MS, by low energy collision-induced dissociation of the derivatized methionine side chain, resulting in the formation of a single characteristic product ion. Structural characterization of identified peptides is then achieved by automatically subjecting the characteristic neutral loss product ion to further dissociation by data dependent product ion scan mode pseudo MS3 under higher collision energy conditions. Quantitative analysis is achieved by measurement of the abundances of characteristic product ions formed by sequential neutral loss scan mode MS/MS experiments from “light” (12C) and “heavy” (13C) stable isotope encoded fixed-charge derivatized peptides. In contrast to MS-based quantitative analysis strategies, the neutral loss scan mode MS/MS method employed here was able to achieve accurate quantification for individual peptides at levels as low as 100fmol and at abundance ratios ranging from 0.1 to 10, present within a complex protein digest.
Evaluation of in vitro storage properties of prestorage pooled whole blood–derived platelets suspended in 100 percent plasma and treated with amotosalen and long‐wavelength ultraviolet light

BACKGROUND: Amotosalen, a psoralen, has been utilized for photochemical treatment (PCT) of apheresis platelets (PLTs) and pooled buffy coat PLTs suspended in additive solution. In the United States, the source of many PLT transfusions is from whole blood–derived PLTs prepared by the PLT‐rich plasma (PRP) method. This study investigated the in vitro PLT properties of amotosalen‐PCT of leukoreduced pools of PLTs prepared by the PRP method and suspended in 100 percent plasma.
Pharmacological actions of aconitine alkaloids.

Aconitine (AC), mesaconitine (MA), hypaconitine (HA) (50 μg/kg i.v.), benzoylaconine (BA), benzoylmesaconine (BM) and benzoylhypaconine (BH) (5 mg/kg i.v.) produced a weak temporary hypotension in rats which was blocked to some extent by atropine. In the isolated guinea pig right atria, MA and HA (3×10-8 g/ml) mediated a positive inotropic action and a positive chronotropic action. At the higher concentration of 10-7 g/ml, AC, MA and HA exhibited the above actions followed by inhibition of contractions and disorder of the beating rate. In the isolated guinea pig ileum, AC, MA and HA (>10-6 g/ml) caused a contraction which was completely blocked by atropine. In the isolated guinea pig vas deferens, AC, MA and HA (>10-5 g/ml) elicited a contraction which was completely obliterated by phentolamine. In the isolated guinea pig hypogastric nerve-vas deferens, the isolated rabbit mesenteric nerve-jejunum and the isolated rat phrenic nerve-diaphragm, responses induced by the electrical stimulation of the corresponding nerves were inhibited by AC, MA and HA (10-6-10-5 g/ml). Some of the mechanisms of the above actions induced by these alkaloids are discussed. These alkaloids more or less potentiated the hexobarbital anesthesia, inhibited the revolution of the wheel cage, and reduced the rectal temperature in mice, indicating that they have a depressant activity on the central nervous system.