Cytosporone E: racemic synthesis and preliminary antibacterial testing

The antibiotic cytosporone E (isolated from the broth of the endophytic fungi CR 200 (Cytospora sp.) and CR 146 (Diaporthe sp.)) was synthesized as a racemic mixture. The key step in the synthesis is the Meyers ortho-alkylation of a chiral aromatic oxazoline. Preliminary antibiotic activity shows antibiosis against Gram-positive bacteria but not Gram-negative bacteria as previously reported.     

阪崎肠杆菌能力验证样品均匀性和稳定性的研究

目的制备出以脱脂奶粉为基质的均匀性好、稳定性强的阪崎肠杆菌(Enterobacter sakazakii)能力验证样品。方法研究奶粉基质的粒度、奶粉和菌粉的混合比例,得到阪崎肠杆菌能力验证样品的最佳均匀性条件;研究包装形式及贮存温度对样品稳定性的影响,得到阪崎肠杆菌能力验证样品的最佳稳定性条件。结果要保证样品足够均匀,奶粉最佳粒径范围为120μmD180μm,1 g菌粉最多与300 g奶粉进行混合;贮存温度对样品稳定性有较大影响,高温明显降低样品稳定性;真空包装可以显著提高样品稳定性。结论以奶粉为基质的阪崎肠杆菌能力验证样品的制备能够更准确地考查乳品微生物检验人员的检测水平,为国内微生物能力验证水平的提高奠定了基础。     

HPV E6/E7 mRNA testing is more specific than cytology in post-colposcopy follow-up of women with negative cervical biopsy

Background

In Norway, women with negative or low-grade cervical biopsies (normal/CIN1) are followed up after six months in order to decide on further follow-up or recall for screening at three-year intervals. A high specificity and positive predictive value (PPV) of the triage test is important to avoid unnecessary diagnostic and therapeutic procedures whereas a low risk of high-grade disease among triage negative women assures safety.

Materials and Methods

At the University Hospital of North Norway, cytology and the HPV mRNA test PreTect HPV-Proofer, detecting E6/E7 mRNA from HPV types 16, 18, 31, 33 and 45, are used in post-colposcopy follow-up of women with negative or low-grade biopsy. In this study, women with negative biopsy after high grade cytology (ASC-H/HSIL) and/or positive HPV mRNA test in the period 2005–2009 were included (n = 520). Histologically confirmed cervical intraepithelial neoplasia of grade 2 or worse (CIN2+) was used as study endpoint.

Results

Of 520 women with negative or low-grade biopsy, 124 women (23.8%) had CIN2+ in follow-up biopsy. The sensitivity and specificity of the HPV mRNA test were 89.1% (95% CI, 80.1–98.1) and 92.5% (95% CI, 88.2–96.7), respectively. The ratios of sensitivity, specificity and PPV of HPV mRNA testing compared to repeat cytology for finding CIN2+ was 1.05 (95% CI: 0.92–1.21), 1.21 (95% CI: 1.12–1.32), and 1.49 (95% CI: 1.20–1.86), respectively. The PPV of mRNA was 77.3% (95% CI, 59.8–94.8) in women aged 40 or older.

Conclusion

Women with negative cervical biopsy require follow-up before resumption of routine screening. Post-colposcopy HPV mRNA testing was as sensitive but more specific than post-colposcopy cytology. In addition, the HPV mRNA test showed higher PPV. A positive mRNA test post-colposcopy could justify treatment in women above 40 years.     

Sensitivity of different E. coli and Salmonella strains in mutagenicity testing calculated on the basis of selected literature

Two microbial screening test systems for gene (point) mutations, the Salmonella typhimurium assay (TA1535, TA1537, TA1538, TA98 and TA100) and the Escherichia coli WP2 reverse-mutation system (WP2, WP2uvrA, WP2pKM101 and WP2uvrApKM101), were compared with regard to sensitivity toward a broad spectrum of compounds that cause base-pair or frameshift mutations and that have known carcinogenic qualities. Based on available published literature we found that all 44 carcinogens and 9 non-carcinogens examined in both test systems also met with criteria for data acceptance drawn up by us. The results obtained are: firstly, that the Salmonella assay is decidedly better validated than the E. coli WP2 test; and secondly, that the E. coli test system sensitivity (91%) is fully on a par with the sensitivity of the Salmonella assay (72%). This last is in divergence from earlier reports, e.g. Brusick et al. (1980), and this difference must be ascribed to the new plasmid-containing strains. The many compounds not tested in the E. coli department result in fewer false negatives in the E. coli test system and their omission constitutes a bias in favour of the E. coli assay. By eliminating compounds that are negative in Salmonella and dropped from the WP2 analysis owing to insufficient data, the sensitivity of the Salmonella system is raised to 84% as compared with 91% for the WP2 assay. The results further indicate that some of the tester strains are superfluous, and show an exceedingly sensitive test can be performed by combining the best tester strains from the two test systems.     

Skin testing versus radioallergosorbent testing for indoor allergens

BACKGROUND: Skin testing (ST) is the most common screening method for allergy evaluation. Measurement of serum specific IgE is also commonly used, but less so by allergists than by other practitioners. The sensitivity and specificity of these testing methods may vary by type of causative allergen and type of allergic manifestation. We compared ST reactivity with serum specific IgE antibodies to common indoor allergens in patients with respiratory allergies. METHODS: 118 patients (3 mo-58 yr, mean 12 yr) with allergic rhinitis and/or bronchial asthma had percutaneous skin testing (PST) supplemented by intradermal testing (ID) with those allergens suspected by history but showed negative PST. The sera were tested blindly for specific IgE antibodies by the radioallergosorbent test (Phadebas RAST). The allergens were D. farinae (118), cockroach (60), cat epithelium (90), and dog epidermal (90). Test results were scored 0-4; ST >/= 2 + and RAST >/= 1 + were considered positive. RESULTS: The two tests were in agreement (i.e., either both positive or both negative) in 52.2% (dog epidermal) to 62.2% (cat epithelium). When RAST was positive, ST was positive in 80% (dog epidermal) to 100% (cockroach mix). When ST was positive, RAST was positive in 16.3% (dog epidermal) to 50.0% (D. farinae). When RAST was negative, ST was positive in 48.5% (cat epithelium) to 69.6% (D. farinae). When ST was negative, RAST was positive in 0% (cockroach) to 5.6% (cat epithelium). The scores of ST and RAST showed weak to moderate correlation (r = 0.24 to 0.54). Regardless of history of symptoms on exposure, ST was superior to RAST in detecting sensitization to cat epithelium and dog epidermal. CONCLUSION: For all four indoor allergens tested, ST was more sensitive than RAST. When both tests were positive, their scores showed poor correlation. Sensitizations to cat epithelium and dog epidermal are common, even in subjects who claimed no direct exposure.     

Interference testing

* Interference occurs when a substance or process falsely alters an assay result. * Interferences are classified as endogenous or exogenous. Endogenous interference originates from substances present in the patient's own specimen. Exogenous interferences are substances introduced into the patient's specimen. * To perform interference studies, proper planning is required. * Interference from haemolysis, icterus and lipaemia are most frequently studied. Haemolysis affects more analytes than does any other type of interference. * Protein interferences are most often associated with paraproteins and predominantly with IgM or IgG and rarely with IgA. * Drug interference may be due to the parent drug, metabolite(s) or additives in the drug preparation. * Collection tube components can affect determination of analytes. * Carryover interference typically occurs when analyte from a high concentration sample (or reagent) is incompletely removed by the analytical system's washing process, whether probe, mixer or cuvette washing. * Immunoassay interferences are most commonly due to antibodies (generally polyclonal). They may be autoantibodies (e.g. in thyroid disease) or heterophile antibodies that predominantly interfere in two-site immunometric (sandwich) assays, forming a bridge between capture and detection antibodies. * Determining if interference is significant requires deviation limits from the original result. * Once interferences are identified during method evaluation or in general use, there is a need to establish procedures for handling affected results as part of the quality system.     

A combined testing protocol approach for mutagenicity testing.

The antischistosomal agent, hycanthone methanesulfonate (HMS), was employed to illustrate the utility of carrying out several mutagenicity tests in a single concurrent animal experiment. Several commonly used procedures that were successfully integrated into a multiple testing protocol included (1) metaphase analysis in bone marrow, (2) micronucleus test in bone marrow, (3) analysis of the urine for mutagenic constituents, and (4) the host-mediated assay using Salmonella typhimurium. In addition to these animal studies, in vitro mutagenicity testing with and without activation was carried out using S. typhimurium. HMS produced positive, dose--response effects in in vitro tests, metaphase analysis, micronucleus test, and urine analysis, but not in the host-mediated assay. The results of these integrated techniques suggest that such a protocol may be a benefit to those concerned with mutagenicity testing of chemicals.     

Biaxial testing of membrane biomaterials: testing equipment and procedures

A testing facility for measuring the biaxial mechanical properties of highly deformable membranes is described. Forces are applied, via strain-gauge force transducers, to four points on each side of an initially square 12 to 25 mm membrane sample to produce biaxial extensions of up to 80 percent of undeformed length. Strain is estimated from the displacement of markers bounding a 1 to 2 mm central square. The accuracy of stress and strain field measurements has been assessed by finite element analysis of a biaxially-loaded isotropic elastic membrane. Major advantages of the present system over those previously described in the literature are that 1) sample mounting procedures are simplified, 2) there is provision for independent adjustment of stress field uniformity and measurement of the applied point forces and 3) faster strain rates can be imposed on the relatively small samples tested.