Miracle-dz/UniFAR-FTUs huggingface dataset:
Rapid Method for Detection of Gram-Positive and -Negative Bacteria in Milk from Cows with Moderate or Severe Clinical Mastitis

ABSTRACT A rapid method for demonstration of gram-positive and gram-negative bacteria in milk is described. The technique is based on dilution of the sample in a medium, followed by filtration through a porous polysulfone membrane with a pore size retaining and concentrating bacteria from the sample. The bacteria concentrated on the surface of the membrane are stained with a cationic dye (toluidine blue) that can be visualized by the naked eye. After staining, the membrane is treated with ethanol-acetic acid (pH 2.8 to 3.0), which causes decolorization of gram-negative bacteria, whereas gram-positive bacteria retain the stain. The method does not require heat fixation, electrical power, microscopic examination, or specially trained personnel. The time needed to perform the test is approximately 5 min. The technique was applied to artificially infected milk and milk from cows with moderate or severe clinical mastitis for detection and differentiation of bacteria. The sensitivity of the filtration method was 92 and 100% for gram-positive and gram-negative bacteria, respectively, compared with traditional bacteriological culture of milk samples. The detection limit was 5 × 106 CFU/ml for Staphylococcus aureus and 1 × 106 CFU/ml forEscherichia coli in spiked milk samples. The overall specificity of the method was 86%. This diagnostic method can provide on-site guidance to the veterinarian to optimize use of antibiotics in mastitis therapy.
A novel HTTP botnet traffic detection method

In this paper we propose a novel technique for detecting HTTP botnet traffic “N-gram based HTTP bot traffic detection” that makes use of Deep Packet Inspection (DPI) of network packets to detect hosts infected with a bot. Botnet is a collection of compromised hosts or computers (bots) which are remotely controlled by its originator (botmaster) under a common command and control (C&C) infrastructure. The proposed botnet detection technique is capable of detecting bots that use encryption to hide the contents of their communication which most of the existing DPI based techniques fail to detect. It is also capable of detecting Fast Flux botnets in which C&C dynamically changes its IP address which most of the existing techniques based on traffic flow analysis fail to detect. The proposed technique is based on the fact that the C&C responds with similar communication patterns with only slight modifications to an HTTP GET request made by a bot. The communications patterns do not vary unless the bot is updated. Performance observations using our proposed botnet detection technique show high accuracy of botnet detection.
Laser capture microdissection of fluorescently labeled embryonic cranial neural crest cells

Summary: This study is the first to report a unique genetic strategy to permanently label mammalian neural crest cells (NCC) with a fluorescent marker, selectively isolate the labeled NCC or their derivatives during murine ontogenesis by laser capture microdissection (LCM), and prepare molecular components, such as RNA, for selective gene expression analyses. Through utilization of a Cre recombinase/loxP system, a genetic strategy that has been used repeatedly to achieve tissue‐specific activation of reporter transgenes in mice, a novel two‐component mouse model was created in which neural crest cells (and their progeny) are indelibly marked throughout the pre‐ and postnatal lifespan of the organism. To generate this mouse model, a Wnt1‐Cre transgenic line was crossed with a mouse line expressing a conditional reporter transgene (“floxed” enhanced green fluorescent protein). Resulting offspring, expressing both the Wnt1‐Cre and “floxed” EGFP alleles, demonstrated EGFP expression in the NCC and all of their derivatives throughout embryonic, postnatal, and adult stages. In the present study, EGFP‐labeled cranial NCC from the first branchial arch of gestational day 9.5 murine embryos were visualized in frozen tissue sections and isolated by LCM under epifluorescence optics. RNA was extracted from “captured” cells and amplified by double‐stranded cDNA synthesis and in vitro transcription. Amplified mRNA samples from “captured” cells were evaluated by TaqMan quantitative, real‐time PCR for the expression of a panel of NCC gene markers. The molecular genetic strategy delineated in this report will facilitate future embryo‐genomic and ‐proteomic analyses of mammalian NCC that will serve to further our understanding of these pluripotent embryonic progenitor cells. genesis 39:58–64, 2004. © 2004 Wiley‐Liss, Inc.
TNFα-Induced Long Pentraxin PTX3 Expression in Human Lung Epithelial Cells via JNK

Long pentraxin 3 (PTX3), an acute-phase protein, is a newly clarified mediator for innate immunity and inflammation. As a soluble pattern recognition receptor, it has a nonredundant role in antifungal infection. Overexpression of PTX3 worsens acute lung injury. The lung epithelium is a critical factor in defense against pulmonary pathogens; it is also involved in acute inflammatory responses related to tissue injury. However, very little is known about how PTX3 is regulated in the lung epithelium. In this study, we found that i.v. injection of LPS induced PTX3 expression in rat lung alveolar epithelium. Using human lung cell lines and primary epithelial cells, we found that PTX3 expression was significantly up-regulated by TNF-α in a time- and dose-dependent manner, but not by LPS. Pretreatment with either actinomycin D or cycloheximide abolished TNF-α-induced PTX3 expression, indicating the requirement for both transcriptional and translational regulation. The TNF-α-induced PTX3 expression was blocked by SP600125, a JNK-specific inhibitor, but not by the inhibitors against NF-κB, ERKs, or p38 MAPK. Knockdown of either JNK1 or JNK2 with small interfering RNA also significantly reduced the regulated PTX3 expression. Thus, lung epithelial cells appear to be a major local source for PTX3 production, which could be induced in vivo from these cells by LPS or other inflammatory stimuli, and may be an important mediator for host defense and tissue damage. The importance of the JNK pathway for the regulated PTX3 expression may be a potential target for its regulation in the lung.
RISE AND FALL OF SYNTHETIC CDO MARKET: LESSONS LEARNED

This paper uses a unique data set of more than 1000 synthetic Collateralized Debt Obligations (CDOs) deals to describe typical structures, their pricing and performance with the aim of identifying the factors behind the spectacular collapse of this important segment of structured credit market in late 2008. The data suggests that mark-to-market losses on many synthetic CDO tranches were much more significant than in case of simpler, lower-rated products despite the former experiencing little or no impairment of the notional. The losses were driven instead by the concentration of relatively limited number of defaults in a short period of time, suggesting that pre-crisis pricing must have seriously underestimated such risk of default clustering. In view of the post-crisis pick-up in synthetic CDO issuance, the paper attempts to heed this lesson and offer a simple factor model of default correlation in the spirit of Marshall–Olkin that is naturally suited to capturing the temporal dimension of default dependencies that have been crucial for synthetic CDOs investors. The model allows building a rich dependence structure capable of consistently fitting standardized iTraxx and CDX index tranches, which makes it ideal for pricing bespoke CDOs.
CD137 on inflamed lymphatic endothelial cells enhances CCL21‐guided migration of dendritic cells

CD137/TNFR9/41BB was originally described as a surface molecule present on activated T and NK cells. However, its expression is broader among leukocytes, and it is also detected on hypoxic endothelial cells and inflamed blood vessels, as well as in atherosclerotic lesions. Here, we demonstrate that lymphatic endothelial cells (LECs) up‐regulate CD137 expression from undetectable baseline levels on stimulation with TNF‐α, LPS, and IL‐1β. CD137 cross‐linking with an agonistic mAb results in NF‐κB nuclear translocation, followed by up‐regulation of VCAM and a 3‐fold increase in the production of the chemokine CCL21. Accordingly, there is a 50% increase in CCR7‐dependent migration toward conditioned medium from activated LECs on CD137 cross‐linking with the agonistic mAb or the natural ligand (CD137L). Such an enhancement of cell migration is also observed with monocyte‐derived dendritic cells transmigrating across CD137‐activated LEC monolayers. Using explanted human dermal tissue, we found that inflamed skin contains abundant CD137+ lymphatic vessels and that ex vivo incubation of explanted human dermis with TNF‐α induces CD137 expression in lymphatic capillaries. More interestingly, treatment with CD137 agonistic antibody induces CCL21 expression and DC accumulation close to lymphatic vessels. Collectively, our results demonstrate that the inflammatory function of lymphatic vessels can be regulated by CD137. —Teijeira, A. , Palazón, A. , Garasa, S. , Marré, D. , Aubá, C. , Rogel, A. , Murillo, O. , Martínez‐Forero, I. , Lang, F. , Melero, I. , Rouzaut, A. CD137 on inflamed lymphatic endothelial cells enhances CCL21‐guided migration of dendritic cells. FASEB J. 26, 3380–3392 (2012). www.fasebj. org
Brain-derived neurotrophic factor gene polymorphisms and schizophrenia: a meta-analysis

Objectives To evaluate whether the G196A and C270T polymorphisms of the brain-derived neurotrophic factor gene are associated with increased risk of schizophrenia. Methods A meta-analysis of nine genetic association studies was carried out. The meta-analysis included genotype data on 1404/1597 schizophrenics/controls for G196A and 877/989 schizophrenics/controls for C270T. Results The overall analysis for investigating the association of the G196A allele G and the risk of developing schizophrenia relative to the allele A, showed significant evidence of heterogeneity (P=0.05, I2=58%) between the studies and nonsignificant association [random effects odds ratio 1.08 and 95% confidence interval (0.88–1.32)]. In Caucasians, there was a trend towards heterogeneity (P=0.19, I2=40%), then, the random and fixed effects odds ratios were 1.24 (0.96–1.60) and 1.27 (1.06–1.53), respectively. For the C270T polymorphism, overall, there was significant evidence of heterogeneity between studies (P=0.07, I2=55%) and the allele T was associated with a 63% increased risk of schizophrenia compared with C allele [random effects odds ratio 1.63 (1.01–2.65)]. The dominant model for allele T produced significant association [random effects odds ratio 1.68 (1.02–2.79)]. No source of bias was seen in the selected studies and the differential magnitude of effect in large versus small studies was not significant. Conclusions The meta-analysis results provided a weak evidence of association between C270T polymorphism and schizophrenia, and large heterogeneity between studies, whereas there was no evidence of association for G196A polymorphism. The above findings reinforce the need for large and more rigorous association studies.
Cell cycle arrest and apoptosis induced by SART-1 gene transduction.

UNLABELLED The biological function of the SART-1 gene product is demonstrated and its potential as a target for cancer gene therapy is discussed. MATERIALS AND METHODS The SART-1 gene was transduced by a recombinant adenovirus vector and its expression was promoted by a CMV promoter. RESULTS The transduction efficiency by recombinant adenoviruses in A549 and MCF-7 cells was determined using a vector expressing luciferase, which showed high expression in the cells. Cell count analysis using Trypan-Blue dye exclusion showed that SART-1 gene transduction inhibited cell growth. Flow cytometry analysis suggested that SART-1 gene transduction induced cell cycle arrest followed by apoptosis. Western blot analysis confirmed that the apoptosis pathway was activated by SART-1 gene transduction. CONCLUSION These results show that SART-1 gene transduction induces cell cycle arrest leading to apoptosis and suggest the possibility of gene therapy against cancer. In addition, SART-1 is known to be a tumor antigen in a range of cancers recognized by T cells, thus a potential strategy would be the combination of suicide gene therapy with immuno-gene therapy.
Motivations, Challenges, and Opportunities of Successful Solvers on an Innovation Intermediary Platform

The study aims is to identify motivations, challenges, and opportunities of successful solvers participating in virtual teams of innovation contests (ICs) organized by an innovation intermediary. Based on 82 interviews of successful solvers, it provides novel insights into ICs. The main motivational factors of successful solvers engaged in problem solving are money, learning, fun, sense of achievement, passion, and networking. Major challenges solvers face include unclear or insufficient problem description, lack of option for communication, language barrier, time zone differences, difficulties in finding suitable team members, framing the results, and difficulties in becoming quick learners and team players. Despite challenges, solvers have many opportunities, such as diversified knowledge, learning culture, developing a different way of thinking, gaining insights from other experts, the ability to work in a diverse environment, options of work after retirement and from distant locations, and a new source of income.
Calibration of X-ray radiographs and its feasible application for 2D/3D reconstruction of the proximal femur

In the past ten years, a variety of computer-assisted surgical technologies have been introduced for total hip replacement (THR). Despite the different approaches, orthopedic surgeons routinely still acquire 2D X-ray radiographs for preoperative diagnostic and templating purposes. Such images provide detailed information in high resolution about the underlying anatomy. But lack of knowledge about the magnification factor of the bony structures complicates the correct interpretation of the real anatomy and therefore could lead to mismatching implants intraoperatively based on the preoperative performed planning. Therefore we developed a calibration device, which can be mounted within the usual clinical environment and allows to improve the accuracy of conventional radiographs by applying a specific calibration algorithm. Furthermore, we investigated the feasibility of reconstructing the surface of the proximal femur on the basis of two calibrated X-ray radiographs. The accuracy of this approach has been thoroughly validated using 14 dry cadaver femur bones (pathological as well as non-pathological cases), one plastic femur bone and one wet cadaver pelvis.
Specific Association Of CD63 with the VLA-3 and VLA-6 Integrins (*)

We screened monoclonal antibodies to cell-surface proteins and selected an antibody, called 6H1, that recognizes a putative integrin-associated protein. The 6H1 monoclonal antibody (mAb) indirectly coprecipitated α3β1 and/or α6β1, but not α2β1, or α5β1 from Brij 96 detergent lysates of multiple cell lines. Large scale purification using the 6H1 mAb yielded a single protein of 45-60 kDa with an amino-terminal sequence that exactly matched CD63. Confirming that the 6H1 mAb recognized the CD63 protein, 6H1 and a known anti-CD63 mAb yielded identical coprecipitation results and identical colocalization into lysosomal granules containing cathepsin D. Furthermore, we used an established anti-CD63 mAb to detect this protein in an α3β1 immunoprecipitate, and also we observed VLA-3 and CD63 colocalization in cellular “footprints. ” Notably, the cytoplasmic domain of α3 was neither required nor sufficient for CD63 association, suggesting that it occurred elsewhere within the α3β1 complex. Knowledge of these specific CD63-α3β1 and CD63-α6β1 biochemical associations should lead to critical insights into the specialized functions of α3β1, α6β1, and CD63.
Embedding Psychosocial Perspectives Within Clinical Management of Low Back Pain: Integration of Psychosocially Informed Management Principles Into Physical Therapist Practice—Challenges and Opportunities

As the biopsychosocial model of health has become increasingly understood, it has become clear that there are complex, interdependent relationships between the physical and biomedical features of low back pain and the psychological and social factors that present concomitantly. Epidemiological studies have not only highlighted that psychological and social factors are associated with back pain and disability but also have shed light on the way in which these factors serve as prognostic indicators, or obstacles to recovery, predicting which patients will have a poor prognosis. Integrating the assessment of these obstacles to recovery into physical therapist practice and using this information to guide clinical decision making have the potential to improve the quality of care offered by physical therapists by improving the targeting of treatments to individuals and enhancing the therapist-patient relationship and adherence to management advice and treatment programs. In turn, such approaches may improve both patients' clinical outcomes and the efficiency and effectiveness of service provision, helping direct interventions to those who need them. This article summarizes the key challenges to embedding psychosocial perspectives within physical therapist practice for patients with low back pain and the opportunities that could be realized by doing so, and it highlights new developments in research, clinical practice, and education that are shaping future directions in this field.
A New Non-Parametric Matching Method for Bias Adjustment with Applications to Economic Evaluations

In health economic studies that use observational data, a key concern is how to adjust for imbalances in baseline covariates due to the non-random assignment of the programs under evaluation. Traditional methods of covariate adjustment such as regression and propensity score matching are model dependent and often fail to replicate the results of randomized controlled trials. We demonstrate a new non-parametric matching method, Genetic Matching, which is a generalization of propensity score and Mahalanobis distance matching (Sekhon forthcoming), using two contrasting case studies. In the first, an economic evaluation of a clinical intervention (Pulmonary Artery Catheterization), applying Genetic Matching to observational data replicates the substantive results of a corresponding randomized controlled trial unlike the extant literature. And in the second case study evaluating capitation versus fee-for-service, Genetic Matching radically improves balance on baseline covariates and overturns previous conclusions based on traditional methods.
Liver-specific mRNA for Insig-2 down-regulated by insulin: Implications for fatty acid synthesis

Insig-1 and -2 are closely related proteins of the endoplasmic reticulum (ER) that block proteolytic activation of sterol regulatory element-binding proteins (SREBPs), transcription factors that activate the synthesis of cholesterol and fatty acids in liver and other organs. When cellular cholesterol levels are high, Insig proteins bind and trap SREBP cleavage-activating protein (SCAP), retaining it in the ER and preventing it from escorting SREBPs from ER to the site of proteolytic activation in the Golgi complex. Here, we report the discovery of a liver-specific transcript of Insig-2, designated Insig-2a. This transcript and the ubiquitous transcript, designated Insig-2b, differ through the use of different promoters that produce different noncoding first exons that splice into a common second exon. Although the Insig-2a and -2b mRNAs encode identical proteins, they differ in patterns of regulation. Insig-2a is the predominant transcript in livers of fed animals, and it is selectively down-regulated by insulin. Insig-2a mRNA increases when mice are fasted, and it declines when they are refed. The transcript also increases in livers of rats whose insulin-secreting pancreatic beta cells have been destroyed by streptozotocin, and it is reduced when insulin is injected. The insulin-mediated fall in Insig-2a may allow SREBP-1c to be processed, thereby allowing insulin to stimulate fatty acid synthesis, even under conditions in which hepatic cholesterol levels are elevated.
Barriers to enterprise development in the Caribbean

ABSTRACT Caribbean economies have suffered from stagnant growth since the 1990s. This can be a feature of small developing economies and is a major concern for policymakers. In this article, we examine establishment-level data to gain a better understanding of the factors that constrain the growth of businesses in the region. In addition to documenting broad differences in obstacles to business within and across the region, we particularly focus on the main obstacles affecting small and medium-sized firms. The econometric analysis highlights three main barriers at a regional level: an inadequately educated workforce; access to finance; and crime, theft and disorder. However, there are variations at the country level and the analysis indicates clusters of countries that experience obstacles to similar degrees. The article concludes with recommendations for alleviating the constraints to enterprise development and in stimulating economic growth.
Microneedles permit transdermal delivery of a skin-impermeant medication to humans

Drugs with poor oral bioavailability usually are administered by hypodermic injection, which causes pain, poor patient compliance, the need for trained personnel, and risk of infectious disease transmission. Transdermal (TD) delivery provides an excellent alternative, but the barrier of skin's outer stratum corneum (SC) prevents delivery of most drugs. Micrometer-scale microneedles (MNs) have been used to pierce animal and human cadaver skin and thereby enable TD delivery of small molecules, proteins, DNA, and vaccines for systemic action. Here, we present a clinical study of MN-enhanced delivery of a medication to humans. Naltrexone (NTX) is a potent mu-opioid receptor antagonist used to treat opiate and alcohol dependence. This hydrophilic and skin-impermeant molecule was delivered from a TD patch to healthy human subjects with and without pretreatment of the skin with MNs. Whereas delivery from a standard NTX TD patch over a 72-h period yielded undetectable drug plasma levels, pretreatment of skin with MNs achieved steady-state plasma concentrations within 2 h of patch application and were maintained for at least 48 h. The MNs and NTX patch were well tolerated with mild systemic and application site side effects. The MN arrays were painless upon administration and not damaged during skin insertion, and no MNs were broken off into the skin. This human proof-of-concept study demonstrates systemic administration of a hydrophilic medication by MN-enhanced TD delivery. These findings set the stage for future human studies of skin-impermeant medications and biopharmaceuticals for clinical applications.