Detection of Salmonella spp. in retail meat products: a comparison between a discontinued commercial kit and a laboratory-developed screening method.

The National Antimicrobial Resistance Monitoring System for Enteric Bacteria retail food surveillance programme screens retail meat samples for the presence of Salmonella spp. to track antimicrobial resistance in food. In this study, a laboratory developed real-time PCR assay that detects Salmonella spp. was evaluated as a screening method to replace the discountinued 3M TECRA kit. The 3M TECRA kit was a commercially available, visual immunoassay used to screen food samples for the presence of Salmonella spp. This kit was discontinued in September 2016 by the manufacturer and an alternative screening method was needed to replace the discontinued TECRA kit. Salmonella spp. is detected by the real-time PCR assay earlier in the screening process than by the TECRA kit. Salmonella spp. can also be reliably isolated from the enrichment broth earlier in the protocol. Additionally, cost analysis shows that the real-time PCR assay saves $2·50 per sample. New York State Department of Health currently uses this real-time PCR assay as a screening method for the presence of Salmonella spp. in retail meat samples. The assay allows for continued monitoring of antimicrobial resistance in Salmonella spp., while providing a cost savings and a decrease in turnaround time. SIGNIFICANCE AND IMPACT OF THE STUDY: The National Antimicrobial Resistance Monitoring System for Enteric Bacteria (NARMS) tracks antimicrobial susceptibility of enteric bacteria in people, food and animals (https://www.fda.gov/animalveterinary/safetyhealth/antimicrobialresistance/nationalantimicrobialresistancemonitoringsystem/). The New York State Department of Health (NYSDOH) became a NARMS retail food surveillance (RFS) site in 2003. The NARMS-RFS programme screens retail meat samples from grocery stores in the United States for the presence of Salmonella spp. and other enteric pathogens to monitor the prevalence of antimicrobial resistance among these pathogens. The NYSDOH developed a rapid and cost-effective real-PCR assay to screen for Salmonella spp. in retail meat products.

Premarket evaluations of the IMDx C. difficile for Abbott m2000 Assay and the BD Max Cdiff Assay.

Clostridium difficile-associated diarrhea is a well-recognized complication of antibiotic use. Historically, diagnosing C. difficile has been difficult, as antigen assays are insensitive and culture-based methods require several days to yield results. Nucleic acid amplification tests (NAATs) are quickly becoming the standard of care. We compared the performance of two automated investigational/research use only (IUO/RUO) NAATs for the detection of C. difficile toxin genes, the IMDx C. difficile for Abbott m2000 Assay (IMDx) and the BD Max Cdiff Assay (Max). A prospective analysis of 111 stool specimens received in the laboratory for C. difficile testing by the laboratory's test of record (TOR), the BD GeneOhm Cdiff Assay, and a retrospective analysis of 88 specimens previously determined to be positive for C. difficile were included in the study. One prospective specimen was excluded due to loss to follow-up discrepancy analysis. Of the remaining 198 specimens, 90 were positive by all three methods, 9 were positive by TOR and Max, and 3 were positive by TOR only. One negative specimen was initially inhibitory by Max. The remaining 95 specimens were negative by all methods. Toxigenic C. difficile culture was performed on the 12 discrepant samples. True C. difficile-positive status was defined as either positive by all three amplification assays or positive by toxigenic culture. Based on this definition, the sensitivity and specificity were 96.9% and 95% for Max and 92.8% and 100% for IMDx. In summary, both highly automated systems demonstrated excellent performance, and each has individual benefits, which will ensure that they will both have a niche in clinical laboratories.

Successful generation of sperm protein 17 (Sp17)-specific cytotoxic T lymphocytes from normal donors: implication for tumour-specific adoptive immunotherapy following allogeneic stem cell transplantation for Sp17-positive multiple myeloma.

Sperm protein 17 (Sp17) is a highly immunogenic cancer-testis antigen expressed by tumour cells from up to 30% of patients with multiple myeloma (MM). We recently successfully generated Sp17-specific human leucocyte antigen (HLA)-A1 and B27-restricted cytotoxic T lymphocytes (CTLs) from the peripheral blood of a healthy donor. Because CTLs were able to kill HLA-matched fresh myeloma cells, it may be possible to generate and administer myeloma-specific donor T cells to MM patients following allogeneic stem cell transplantation to enhance graft-versus-myeloma (GVM) without inducing graft-versus-host disease (GVHD). To determine how widely applicable this approach is, we have determined the ability to generate Sp17-specific CTLs from four consecutive healthy donors with other HLA class I phenotypes. We found that Sp17-specific HLA class I-restricted CTLs could be easily generated from all four donors. Sp17-specific CTLs were primarily CD8 in phenotype and produced interferon-gamma and very little interleukin-4. These T cells killed target cells primarily via the perforin-mediated route. These results therefore suggest that myeloma-specific donor T-cell infusion that targets Sp17 to selectively enhance GVM could be applicable to patients with Sp17+ MM.

Characteristics of glottis-induced turbulence in oscillatory flow: an empirical investigation.

Turbulence inducement from the glottis was scrutinized by employing an idealized model of the larynx and trachea for oscillatory flow conditions. The characterization of turbulence was achieved with the two-component velocity measurements of split-film probe anemometry and with the flow visualization of a smoke-wire technique. The apertures of two different (triangular and circular) shapes were utilized in the airway model to address the distinct effects of the triangular-shaped glottal aperture on the generation, development, and decay of turbulence. One of the salient turbulence characteristics for the triangular aperture case was found to be the relatively high turbulence levels around the center region (2r/D approximately 0) in conjunction with the asymmetric mean axial velocity across the frontal-rear (A-O-P) plane of the trachea at one tracheal diameter (x/D = 1) downstream from the glottis. The detailed turbulence properties such as the Reynolds shear stresses and turbulence intensities for the triangular aperture case differed significantly from those for the circular aperture case within a few tracheal diameters (x/D < 7) downstream from the apertures. The glottis-induced turbulence was incipient during the acceleration phase of inspiration and convected downstream with the traits of decaying turbulence.

[Accidental poisoning with silo gas]. / Przypadkowe zatrucie gazem silosowym.

Eight cases of poisoning in workers cleaning silo are presented. Silo gas, produced during fermentation of vegetable material, contains very toxic nitrogen oxides. In this group three workers died within silo, four patients were hospitalized (one of them with acute toxic pulmonary oedema, two with sings of pneumonia, one had only transient decrease of consciousness) and recovered without detectable sequelae. One patient, in general good condition, refused hospitalization and recovered.

Correlation of altered tyrosine phosphorylation with methotrexate resistance in a cisplatin-resistant subline of L1210 cells.

Collateral resistance to cisplatin and methotrexate has been reported in several cell lines. A murine leukemia cell line (L1210/DDP) selected for cisplatin resistance also has been shown to be highly resistant to methotrexate. Of the mechanisms proposed for methotrexate resistance, only changes in methotrexate transport into the cells were found in an earlier report. Methotrexate enters mammalian cells via an active transport system. In the present study, we demonstrated that the transport into the cell may be impaired in the resistant cells due to altered tyrosine phosphorylation of a membrane protein with a molecular mass of 66 kDa. This alteration was manifested by altered tyrosine phosphorylation of the 66 kDa protein and may be an underlying modification that renders the cells resistant to methotrexate. These results suggest involvement of tyrosine phosphorylation in folate transport and methotrexate resistant in L1210/DDP cells.

Investigations on the mechanisms of methotrexate resistance in a cisplatin-resistant L1210 murine leukemia cell subline.

We report a murine leukemia cell variant (L1210/DDP), selected for cisplatin (DDP) resistance, to be cross-resistant to methotrexate (MTX). Cross-resistance of L1210 cells to DDP and MTX has been observed by others, and has also been recorded in P388 murine leukemia and SSC-25 human squamous carcinoma cells. We demonstrated that MTX resistance is not due to dihydrofolate reductase (DHFR) gene amplification, increased DHFR enzyme activity or decreased MTX binding to the target enzyme. Of the mechanisms commonly proposed for MTX resistance, only differences in transport were observed when comparing sensitive (L1210/0) and resistant (L1210/DDP) cells. Our results suggest that MTX resistance in L1210/DDP cells is due to altered methotrexate uptake.

[Low molecular weight heparin (Fraxiparine) as adjunctive therapy with thrombolysis for acute myocardial infarction: a pilot study with a one year follow up]. / Drobnoczasteczkowa heparyna (Fraksyparyna) jako uzupe-nienie leczenia trombolitycznego swiezego zawalu serca; próba "pilotowa" z roczna obserwacja chorych.

In 53 patients with recent (< 6 hrs) acute myocardial infarction a study was undertaken to evaluate the safety of conjunctive therapy with streptokinase (1.5 mln U), aspirin (150 mg) and low molecular weight heparin (Fraxiparine). Patients were treated with Fraxiparine 250 U anti-Xa IC/kg/24 hrs iv for 2 days (with bolus 12.5 U anti-Xa IC/kg), and 125 U anti-Xa IC/kg twice a day sc for 5 subsequent days. Clinical course in one-year observation was compared regarding the time the therapy was initiated. In the group undergoing therapy 3-6 hrs after the infarct had occurred 4 (7.5%) patients died (2 during hospitalization, 2 after discharge). In 31 patients treated within 3 hrs of the myocardial infarction there were fewer cases of recurrent myocardial infarction, unstable angina or congestive heart failure necessitating rehospitalization their (9.1%) than in 22 patients included in the treatment regimen between 3 rd and 6th h of the infarction (27.3%). Earlier thrombolysis was also connected with higher left ventricular ejection fraction (55 +/- 8% vs 49 +/- 10%) and more frequent peak CK-MB values 12 hrs after thrombolysis (81% and 68% of patients respectively). Neither symptomatic deep vein thrombosis nor pulmonary embolism was detected. The left ventricular thrombosis was diagnosed by echocardiography in 4 of 20 patients (20%) with the first anterior myocardial infarction. There was neither bleeding requiring blood transfusion nor cerebrovascular stroke. The treatment with Fraxiparine did not induce the prolongation of APTT values. Conjunctive thrombolytic therapy with low molecular weight heparin was safe and followed by a favorable outcome of the acute myocardial infarction, especially if instituted within the first 3 hrs after the onset of infarction.

A recombinagenic effect of strychnine in Salmonella typhimurium.

The aroC321 allele in Salmonella typhimurium permits a positive selection for genetic duplications. Bacteria that contain a large genetic duplication are detected as tryptophan prototrophs in aroC321 strains and occur at a spontaneous frequency greater than 1/10(4) cells plated on the selection medium. Duplications originate by a recombinational mechanism, and the induction of duplications by chemicals or radiation may therefore be considered to be a recombinagenic effect. We have found that strychnine is a potent recombinagen in this system; it causes a dose-dependent increase in the frequency of genetic duplications, and very high frequencies of duplications are recovered at high doses. In contrast, brucine, the 2,3-dimethoxy derivative of strychnine, caused no increase in duplication frequencies under the identical conditions.

Induction of neonatal rat skin and liver aryl hydrocarbon hydroxylase by coal tar and its constituents.

Topical application of coal tar solution (USP) to neonatal rats resulted in the induction of skin and liver aryl hydrocarbon hydroxylase (AHH) activities. Furthermore indirect exposure of the animals to coal tar vapors resulted in induction of the enzyme in skin and liver. Cutaneous application of coal tar to pregnant rats resulted in induction of skin and liver AHH activity in both mothers and prenatal rats. Among several defined constituents of coal tar tested benzo(a)pyrene (BP), anthracene and acridine were found to have measurable induction effects on neonatal rat skin and liver AHH. These studies indicate that therapeutic coal tar solution as well as selected defined chemical constituents of coal tar are capable of altering the activity of AHH in skin and liver.