Miracle-dz/UniFAR-FTUs huggingface dataset:
Alielic exchange in Pseudomonas aeruginosa using novel ColE1‐type vectors and a family of cassettes containing a portable oriT and the counter‐selectable Bacillus subtilis sacB marker

An improved method for allele replacement in Pseudomonas aeruginosa was developed. The two main ingredients of the method are: (i) novel ColE1‐type cloning vectors derived from pBR322 and pUC19; and (ii) a family of cassettes containing a portable oriT, the sacB gene from Bacillus subtilis as a counter‐selectable marker, and a chloramphenicol‐resistance gene allowing positive selection of both oriT and sacB. Introduction of plasmid‐borne DNA into the chromosome was achieved in several steps. The DNA to be exchanged was first cloned into the new ColE1‐type vectors. After insertion of the oriT and sacB sequences, these plasmid were conjugally transferred into P. aeruginosa and plasmid integrants were selected. Plating on sucrose‐containing medium allowed positive selection for both plasmid excision and curing since Pseudomonas aeruginosa strains containing the sacB gene in single‐ or multiple copy were highly sensitive to 5% sucrose in rich medium. This procedure was successfully used to introduce an agmR mutation into P. aeruginosa wild‐type strain PA01 and should allow the exchange of any DNA segment into any non‐essential regions of the P. aeruginosa chromosome.
Comparing fludeoxyglucose F18-PET assessment of regional cerebral glucose metabolism and [11C]dihydrotetrabenazine-PET in evaluation of early dementia and mild cognitive impairment.

OBJECTIVE To compare assessment of regional cerebral metabolic changes with [(11)C]dihydrotetrabenazine (DTBZ)-positron emission tomography (PET) measurement of regional cerebral blood flow (K(1)) and fludeoxyglucose F18 (FDG)-PET measurement of regional cerebral glucose uptake (CMR(glc)) in a clinically representative sample of subjects with mild dementia and mild cognitive impairment (MCI). DESIGN [(11)C]Dihydrotetrabenazine-PET K(1) and FDG-PET CMR(glc) measurements were performed. SETTING University-based cognitive disorders clinic. PARTICIPANTS Fifty subjects with either mild dementia (Mini-Mental State Examination score > or = 18) or MCI. Their results were compared with those of 80 normal control subjects. MAIN OUTCOME MEASURES The DTBZ-PET regional K(1) and FDG-PET CMR(glc) measurements were compared with standard correlation analysis. The overall patterns of DTBZ-PET K(1) and FDG-PET CMR(glc) deficits were assessed with stereotaxic surface projections (SSPs) of parametric images. RESULTS The DTBZ-PET regional K(1) and FDG-PET CMR(glc) measurements were highly correlated, both within and between subjects. The SSP maps of deficits in DTBZ-PET regional K(1) and FDG-PET CMR(glc) measurements were markedly similar. The DTBZ-PET K(1) SSP maps exhibited a mild decrease in sensitivity relative to FDG-PET CMR(glc) maps. CONCLUSIONS Both DTBZ-PET K(1) and FDG-PET CMR(glc) measurements provide comparable information in assessment of regional cerebral metabolic deficits in mild dementia and MCI. Blood flow measures can assess regional cerebral metabolism deficits accurately in mild dementia and MCI. Blood flow assessments of regional cerebral metabolic deficits can be combined with tracer binding results to improve utility of PET imaging in mild dementia and MCI.
Demographic Characteristics of Extinction in a Small, Insular Population of House Sparrows in Northern Norway

Abstract: In conservation ecology there is an urgent need for indicators that can be used to predict the risk of extinction of populations. Identifying extinction‐prone populations has been difficult because few data sets on the demographic characteristics of the final stage to extinction are available and because of problems in separating out stochastic effects from changes in the expected dynamics. We documented the demographic changes that occurred during the period prior to extinction of a small island population of House Sparrows (Passer domesticus) after the end of permanent human settlement. A mark‐recapture analysis revealed that this decline to extinction was mainly due to increased mortality after closure of the last farm that resulted in a negative long‐term‐specific growth rate. No change occurred in either the structural composition (breeding sex ratio and age distribution) of the population or in female recruitment. No male, however, recruits were produced on the island after the farm closure. Based on a simple, stochastic, density‐dependent model we constructed a population prediction interval (PPI) to estimate the time to extinction. The 95% PPI slightly overestimated the time to extinction with large uncertainty in predictions, especially due to the influence of demographic stochasticity and parameter drift. Our results strongly emphasize the importance of access to data on temporal variation that can be used to parameterize simple population models that allow estimation of critical parameters for credible prediction of time to extinction.
Phosphorus-31 Nuclear Magnetic Resonance Study of the Effect of Pentachlorophenol (PCP) on the Physiologies of PCP-Degrading Microorganisms

ABSTRACT Free and agarose-encapsulated pentachlorophenol (PCP)-degradingSphingomonas sp. isolates UG25 and UG30 were compared toSphingomonas chlorophenolica ATCC 39723 with respect to the ability to degrade PCP. Pretreatment of the UG25 and UG30 strains with 50 μg of PCP per ml enabled the cells to subsequently degrade higher levels of this environmental pollutant. Similar treatment of ATCC 39723 cells had no effect on the level of PCP degraded by this strain. Phosphorus-31 nuclear magnetic resonance spectra of agarose-immobilized strains UG25 and UG30 grown in the absence of PCP showed that there was marked deenergization of the cells upon exposure to a nonlethal concentration of PCP (120 μg/ml). For example, no transmembrane pH gradient was observed, and the ATP levels were lower than the levels obtained in the absence of PCP. The transmembrane pH gradient and ATP levels were restored once the immobilized cells had almost completely degraded the PCP in the perfusion medium. PCP-pretreated cells, on the other hand, maintained their transmembrane pH gradient and ATP levels even in the presence of high levels of PCP. The ability of PCP-pretreated strain UG25 and UG30 cells to remain energized in the presence of PCP was shown to correlate with an altered membrane phospholipid profile; these cells had a higher concentration of cardiolipin than cells cultured in the absence of PCP. Strain ATCC 39723, which did not degrade higher levels of PCP after PCP pretreatment, did not show this response.
A Preliminary Test of the Interpersonal Psychological Theory of Suicidal Behavior in Young People With a First Episode of Psychosis.

OBJECTIVES The interpersonal-psychological theory (IPT) of suicidal behavior suggests that an individual who does not feel they belong and believes they are a burden combined with the capability to attempt suicide is more likely to attempt suicide. The study aimed to investigate this hypothesis in the context of psychosis. METHOD Young people with a first episode of psychosis (N = 45) who considered suicide, attempted suicide, and had no history of suicide or attempt were compared on self-report measures of suicidal desire, capability for suicide, and substance use. RESULTS No significant differences were found between groups in terms of suicidal desire or capability. All participants perceived they were a burden, did not feel they belonged, and had the capability for suicide. Poisoning (n = 22) was the most frequently reported method of a suicide attempt. Of those who had attempted suicide, 50% (n = 7) reported that it had been related to a psychotic episode. Of the participants, 30 reported previous suicidal ideation mainly to completely end or stop the distress they were experiencing. Levels of depression were significantly different between groups; moderate levels of depression were reported in the suicidal ideation and attempt groups. CONCLUSION The concepts of IPT appear to resonate with the experience of psychosis, regardless of suicidality. Specific features of psychosis and their influence on suicidality are worth further exploration.
Polarized distribution of neutral endopeptidase 24.11 at the cell surface of cultured human intestinal epithelial Caco-2 cells.

The human colon cancer cell line Caco-2 undergoes spontaneous enterocytic differentiation during growth, and expresses a number of brush-border-membrane-associated hydrolases typical of a differentiated phenotype. Among these are alkaline phosphatase, dipeptidyl peptidase IV and sucrase-isomaltase (sucrase, EC 3.2.1.48). Neutral endopeptidase 24.11 [EC 3.4. 24.11, neprilysin (NEP)] is another abundant protease of normal enterocytes but its presence in Caco-2 cells has not been fully documented yet. In this paper, we show that Caco-2 cell extracts hydrolyse tritiated [D-Ala2Leu5]enkephalin with a Km of 180 microM, very close to the value obtained for the NEP present in the rabbit kidney (118 microM). Western-blot analysis of brush-border membranes purified from post-confluent cells revealed a protein with an apparent molecular mass of 94000 Da similar to that of the rabbit kidney NEP. The amount of enzyme in cell extracts increased as a function of the age of the culture, indicating that NEP expression is correlated with the degree of cell differentiation as is also the case for sucrase and dipeptidylpeptidase IV (DPP-IV). Binding of a radiolabelled antibody to Caco-2 cell monolayers grown on semi-permeable filters indicated that 95% of NEP molecules present at the cell surface are on the apical side. Immunocytochemical and flow cytometric analysis of intact and permeabilized cells were also used to investigate the presence of NEP and DPP-IV at the surface of Caco-2 cells. Whereas DPP-IV staining appeared to be homogeneous throughout the entire cell population, NEP-related fluorescence exhibited a bimodal distribution which indicates an uneven expression of the protein at the cell surface. Permeabilization of monolayers with saponin before staining restored a labelling pattern for NEP similar to the one obtained for DPP-IV. This suggests that although DPP-IV and NEP follow similar patterns of expression when enzymic activities are measured on whole-cell extracts, targeting of these brush-border proteins to the cell surface appears to be regulated in different ways.
Adulthood consumption of preserved and nonpreserved vegetables and the risk of nasopharyngeal carcinoma: A systematic review

The incidence rates of nasopharyngeal carcinoma (NPC) are dramatically higher in certain regions of Asia compared to the rest of the world. Few risk factors for NPC are known; however, in contrast to the hypothesized health benefits of nonpreserved vegetables, it is thought that preserved vegetable intake may play a role in contributing to the higher incidence of NPC in high‐risk regions. Therefore, the purpose of this study was to systematically review the epidemiologic evidence on the associations between adulthood intake of preserved and nonpreserved vegetables and NPC risk. A search of the epidemiological literature from 1966 to 2004 was performed using several bibliographic databases, including PubMed and the Chinese Biomedical Literature Database System. There were no language restrictions. Meta‐analysis was conducted to obtain pooled odds ratios (ORs) for the highest‐versus‐lowest categories of preserved and nonpreserved vegetable intake. A total of 16 case‐control studies were identified in the search. Results showed that highest‐versus‐lowest preserved vegetable intake was associated with a 2‐fold increase in the risk of NPC (Random Effects Odds Ratio (RE OR) 2.04; 95% Confidence Limits (CL) 1.43, 2.92). Conversely, high nonpreserved vegetable intake was associated with 36% decrease in the risk of NPC (RE OR 0.64; 95% CL 0.48, 0.85). Findings for both preserved and nonpreserved vegetables were consistent across vegetable type and by country of study. Further research in high‐risk areas to gain insight into the risk associated with preserved vegetables and protection associated with nonpreserved vegetables may advance understanding of NPC and yield clues for prevention. © 2006 Wiley‐Liss, Inc.
A Cost-effectiveness Analysis of Cyclosporine in Cadaveric Kidney Transplantation

Expected costs and health outcomes associated with cadaveric kidney transplantation using cyclosporine (CsA) plus steroids, azathioprine (Aza) plus steroids, and "dialysis only" were estimated from both a societal perspective and a Medicare perspective. Published data on patient and graft survival and treatment costs were incorporated into a Markov model to predict the ten-year experience of hypothetical cohorts of 1000 35-year-old persons with end-stage renal disease (ESRD) exposed to each treatment option. In the base-case analysis conducted from the societal perspective, ten-year cumulative costs for the "dialysis only," CsA, and Aza cohorts were $181, $147, and $138 million, respectively. Transplantation using CsA rather than Aza would cost an estimated $19,800 per additional life-year and $9,700 per additional graft-year; whereas from Medicare's perspective, CsA would be less costly than Aza. This analysis suggests that under present regulations, widespread use of CsA instead of Aza for cadaveric graft recipients would result in significant cost shifting from Medicare to the private sector; but from a societal perspective, this would result in no, or at worst, relatively inconsequential, additional health expenditures. A policy whereby dollar savings achieved by Medicare from improved graft survival were used to help underwrite the cost of CsA for cadaveric kidney transplants would promote access to this drug and have little impact on the overall cost of care for individuals with ESRD. Key words: cost- effectiveness; cyclosporine; end-stage renal disease; cadaveric kidney transplantation; Med icare. (Med Decis Making 6:199-207, 1986)
Rapid Microwave Fixation of Cell Monolayers Preserves Microtubule-associated Cell Structures

Microwave (MW) fixation has been suggested as a method to rapidly immobilize cellular dynamics for fine structural studies in the electron microscope. To show its suitability for studies on cell monolayers, one has to apply MW fixation systematically in correlation with samples on the light microscopy level. Examples for MW fixation of cell monolayers, however, are still rare. MW-accelerated fixation for relatively long periods of time (1–2 min) has been reported without showing its suitability at the fine structural level. Here, we provide a rapid MW fixation protocol for cell monolayers on a subminute time scale. The impact of the MW-accelerated glutaraldehyde fixation on temperature-sensitive cytoskeletal components such as microtubules was evaluated. For testing the effectiveness of MW-assisted primary fixation, saponin treatment of the monolayers was included. Simultaneous MW-accelerated fixation and extraction by saponin was necessary to achieve a gradual improvement in visualization of cytoskeletal aspects in association with cell junctions, mitochondria, and centrioles. To establish a valuable routine program for fine structural studies of resin-embedded cell models on substrata, a protocol combining MW fixation with automatic processing in a tissue processor is provided.
Melanoma-Reactive CD8 + T Cells Recognize a Novel Tumor Antigen Expressed in a Wide Variety of Tumor Types

An autologous melanoma cell line selected for loss of expression of the immunodominant MART-1 and gp100 antigens was initially used to carry out a mixed lymphocyte tumor culture (MLTC) in a patient who expressed the human leukocyte antigen (HLA)-A1 and HLA-A2 class I major histocompatibility complex alleles. Ten clones identified from this MLTC seemed to recognize melanoma in an HLA-A1–restricted manner but failed to recognize a panel of previously described melanoma antigens. The screening of an autologous melanoma cDNA library with one HLA-A1–restricted melanoma-reactive T-cell clone resulted in the isolation of a cDNA clone called AIM-2 (antigen isolated from immunoselected melanoma-2). The AIM-2 transcript seemed to have retained an intronic sequence based on its alignment with genomic sequences as well as expressed sequence tags. This transcript was not readily detected after Northern blot analysis of melanoma mRNA, indicating that only low levels of this product may be expressed in tumor cells. Quantitative reverse transcriptase–polymerase chain reaction analysis, however, demonstrated a correlation between T-cell recognition and expression in HLA-A1–expressing tumor cell lines. A peptide that was encoded within a short open reading frame of 23 amino acids and conformed to the HLA-A1 binding motif RSDSGQQARY was found to represent the T-cell epitope. The AIM-2–reactive T-cell clone recognized a number of neuroectodermal tumors as well as breast, ovarian, and colon carcinomas that expressed HLA-A1, indicating that this represents a widely expressed tumor antigen. Thus, AIM-2 may represent a potential target for the development of vaccines in patients bearing tumors of a variety of histologies.
Mouse Relapse Model of Clostridium difficile Infection

ABSTRACT Clostridium difficile is the causative agent of primary and recurrent antibiotic-associated diarrhea and colitis in hospitalized patients. The disease is caused mainly by two exotoxins, TcdA and TcdB, produced by the bacteria. Recurrent C. difficile infection (CDI) constitutes one of the most significant clinical issues of this disease, occurs in more than 20% of patients after the first episode, and may be increasing in frequency. However, there is no well-established animal model of CDI relapse currently available for studying disease pathogenesis, prevention, and therapy. Here we report the establishment of a conventional mouse model of recurrence/relapse CDI. We found that the primary episode of CDI induced little or no protective antibody response against C. difficile toxins and mice continued shedding C. difficile spores. Antibiotic treatment of surviving mice induced a second episode of diarrhea, while a simultaneous reexposure of animals to C. difficile bacteria or spores elicited a full spectrum of CDI similar to that of the primary infection. Moreover, mice treated with immunosuppressive agents were prone to more severe and fulminant recurrent disease. Finally, utilizing this model, we demonstrated that vancomycin only delayed disease recurrence, whereas neutralizing polysera against both TcdA and TcdB completely protected mice against CDI relapse. In conclusion, we have established a mouse relapse CDI model that allows for future investigations of the role of the host immune response in the disease's pathogenesis and permits critical testing of new therapeutics targeting recurrent disease.
Solid self-nanoemulsifying drug delivery system (S-SNEDDS) for oral delivery of glimepiride: development and antidiabetic activity in albino rabbits

Abstract Context: This study presents novel self-nanoemulsifying drug delivery system potential of oral delivering which leads poorly aqueous soluble drug glimepiride. Objective: The objective of this study was to prepare solid self-nanoemulsifying drug delivery system (S-SNEDDS) for the improved oral delivery of glimepiride and to evaluate its therapeutic efficacy in albino rabbits. Results and discussion: The droplet size analyses revealed a droplet size of less than 200 nm. The solid state characterization of S-SNEDDS by scanning electron microscopy (SEM), X-ray powder diffraction and differential scanning calorimetry (DSC) revealed the absence of crystalline glimepiride in the S-SNEDDS. The in vitro dissolution studies revealed that the significant improvement in glimepiride release characteristics. The effect of S-SNEDDS on therapeutic efficacy of glimepride was assessed in albino rabbits by monitoring blood glucose levels and compared with free drug suspension, L-SNEDDS. The S-SNEDDS showed significant (p < 0.05) increase in in vitro drug release and therapeutic efficacy as compared with free drug. Conclusion: This study demonstrated that S-SNEDDS is a promising novel drug delivery system of glimepride to enhance oral delivery.
A Lightweight Authenticated Communication Scheme for Smart Grid

In smart grid, various kinds of sensors have been equipped in smart meters to collect real-time consumption data and execute the instructions from the power management center, which leads to a more robust and customer-friendly grid system. However, a smart grid enables the two-way communication between the electricity supplier and users, which brings security challenges to the smart grid. In this paper, we propose a lightweight authenticated communication scheme for the smart grid, which ensures a secure two-way communication between the smart meters and the neighborhood gateway. Since in the proposed scheme, the smart meters only need to execute the bitwise exclusive-OR operations to encrypt collected usage data, and Lagrange interpolation formula is used to authenticate the sender; the transmitted messages can be real-timely authenticated. On the other side, the storage burden and communication cost of the smart meters can be significantly reduced. The security analysis shows that the messages transmission between the smart meters and the neighborhood gateway can be done in a confidential and authenticated manner. Furthermore, compared with other existing schemes, the performance analysis demonstrates that the proposed scheme has both lower storage and communication cost.
Multiple frames of reference: Why international student workers in Australia tolerate underpayment

This article advances research on why international students, who comprise a growing segment of the workforce in many countries, are underpaid. By revisiting Piore’s dual frames of reference theory, the article builds an important knowledge base around migrant workers’ tolerance of low pay. The research uses mixed methods incorporating a survey of 1433 international students, and interviews with 40 of them. Drawing on segmented labour market theory and examining workers’ ‘multiple frames of reference’, the article seeks to explain migrant workers’, in this case international students’, tolerance of extensive and persistent underpayment. While migrants’ reference to lower pay in their home country has long been accepted as explanation for their acceptance of low pay, it is not found to be a significant factor. The new concept of ‘peer frame of reference’ is developed to explain their tolerance of underpayment.