PURPOSE:The purposes of this study were to determine (1) the optimal techniques for and potential diagnostic usefulness of the polymerase chain reaction (PCR) in early Lyme disease, and (2) the true frequency and clinical correlates of PCR-documented blood-borne infection in the dissemination of Lyme disease. PATIENTS AND METHODS:We performed a prospective, controlled, blinded study of PCR, culture, and serology on fractionated blood samples from 105 patients; 76 with physician-diagnosed erythema migrans and 29 controls. Clinical characteristics of the patients were obtained with a standardized data entry form and correlated with results of the laboratory studies. RESULTS:Only 4 of the 76 (5.3%) patients with erythema migrans were culture positive; however, 14 of 76 (18.4%) had spirochetemia documented by PCR of their plasma. None of 29 controls were PCR or culture positive (P = 0.007, versus patients). PCR-documented spirochetemia correlated with clinical evidence of disseminated disease; 10 of 33 patients (30.3%) with systemic symptom(s) were PCR positive compared to 4 of 43 (9.3%) without such evidence (P = 0.02). PCR positivity was more frequent among patients with each of four specific symptoms: fever, arthralgia, myalgia, and headache (all P < 0.05). A higher total number of symptoms (median 2.5 in PCR-positive patients versus 0 in PCR-negative controls; P < 0.01) and the presence of multiple skin lesions (37.5% of patients with multiple, versus 13.3% of patients with single lesions [P = 0.04] were also correlated with PCR positivity. Patients with both systemic symptoms and multiple skin lesions had a 40% PCR-positivity rate; however, 4 of 42 (9.5%) asympatomatic patients with only single erythema migrans lesions were also PCR positive. In multivariate analysis using logistic regression, the number of systemic symptoms was the strongest independent predictor of PCR positivity (P = 0.004). CONCLUSIONS:PCR detection of Borrelia burgdorferi is at least three times more sensitive than culture for identifying spirochetemia in early Lyme disease and may be useful in rapid diagnosis. PCR positivity significantly correlates with clinical evidence of disease dissemination. Bloodstream invasion is an important and common mechanism for the dissemination of the Lyme disease spirochete.

Liver DNA and RNA were isolated from rats treated with the liver carcinogens N-nitrosopyrrolidine (NPYR) and N-nitrosohexamethyleneimine (NHX). After hydrolysis in 70% perchloric acid (100 degrees C, 1.0 h), 70% of the radioactivity in both the DNA and RNA hydrolysates chromatographed as a single peak. The material from both hydrolysates had comparable Rf values on cation exchange and Sephadex G-10 chromatography. Subsequent experiments indicated this material was volatile. After depurination (0.1 M HCl) of the DNA from NPYR- and NHX-treated rats, Sephadex G-10 chromatography separated only a single radioactive peak which co-eluted with the apurinic acid at the void volume. The material which comprised this peak was not volatile. After dialysis of the same 0.1 M HCl hydrolysate from NHX-treated rats, 98% of the radioactivity remained attached to the apurinic acid. These 2 cyclic nitrosamines appear to produce alkylating species that: (1) are capable of extensive, if not exclusive, phosphotriester formation; or (2) have 2 active sites that cross-link to keep purines attached to apurinic acid after 0.1 M HCl hydrolysis.

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTEvidence for the occurrence and formation of diazonium ions in the Agaricus bisporus mushroom and its extractsAllan E. Ross, Donald L. Nagel, and Bela TothCite this: J. Agric. Food Chem. 1982, 30, 3, 521–525Publication Date (Print):May 1, 1982Publication History Published online1 May 2002Published inissue 1 May 1982https://doi.org/10.1021/jf00111a028RIGHTS & PERMISSIONSArticle Views120Altmetric-Citations42LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (541 KB) Get e-Alertsclose Get e-Alerts

A chromatographic technique was developed that could clearly separate beta-N [gamma-L(+)-glutamyl]-4-hydroxymethylphenylhydrazine (agaritine) from all other components in 10-500-microliters samples of mushroom extracts. Locally purchased mushrooms were found to contain mean levels of 0.4 - 0.7 mg agaritine g. The agaritine content of the mushrooms had decreased by 2-47% after 1 wk of storage in a domestic refrigerator and by 36-76% after 2 wk of such storage. Canned mushroom soup and canned mushrooms did not contain detectable agaritine; a sample of frozen mushrooms contained a mean level of 0.33 mg/g and a batch of fresh mushrooms lost about 32% of their agaritine content on cooking. In mice given 3 mg agaritine by gavage, agaritine was detected in all parts of the gastro-intestinal tract 15 min after dosing, but none was detectable in the gut after 3 hr. The enzyme gamma-glutamyltranspeptidase derived from pig's kidney was found to be capable of decomposing agaritine to glutamic acid and 4-(hydroxymethyl)phenylhydrazine, and to have nine times such activity as an enzyme isolated from mushrooms.

Five cyclic nitrosamines, nitrosoazetidine, nitrosopyrrolidine, nitrosopiperidine, nitrosomorpholine, and dinitrosopiperazine, labeled with deuterium and tritium, were injected into rats and then the nucleic acids of the liver were examined for the presence of alkylated bases derived from the nitrosamines. A semiquantitative mass spectrometric method able to detect 25 ng of a base was used. No deuterium-containing alkylated base was detected, although a small radioactive peak was present in the liver RNA from nitrosomorpholine-treated rats. Nitrosomethylcyclohexylamine gave rise to a deuterium-labeled (—CD3) 7-methylguanine in the liver nucleic acids, showing that diazomethane was not an intermediate in its formation, but not in the esophagus nucleic acids, although esophagus is a target organ for this nitrosamine and liver is not. The expected correlation between carcinogenesis by these nitrosamines and alkylation of nucleic acids in vivo was not seen.

Summary Nitrosomethylurea (NMU) and nitrosoethylurea (NEU) were given to rats by injection of 10 and 30 mg, respectively, in 5 ml of 0.9% NaCl solution into the hepatic portal vein. Of 20 animals treated with NMU, 12 developed tumors; 13 of 20 treated with NEU developed tumors; there were tumors in 6 of 20 0.9% NaCl solution controls. Most of the nitrosoalkylurea-treated animals had tumors of more than 1 organ. There seemed to be no predilection for a particular organ, except that 5 NEU-treated animals had brain tumors and 7 had skin tumors; 3 NMU-treated animals had skin tumors. There was no liver cell damage detectable by light microscopy following the treatment. Analysis of liver nucleic acids following similar treatment of rats with deuterium-labeled NMU and NEU showed no detectable alkylation of DNA. However, deuterium-labeled 7-methylguanine was present in the RNA of NMU-treated rats, but no 7-ethylguanine could be detected in the RNA of NEU-treated rats. This difference in ability to alkylate nucleic acids could not be correlated with the similar tumorigenic activity of NMU and NEU. The identification of methyl- d 3 -guanine as the base formed from NMU- d 3 showed that diazomethane was not an intermediate in nucleic acid alkylation by NMU.

The alkylation of nucleic acids of liver, lung, and intestine of the rat in vivo was studied with the use of diethylnitrosamine (DEN), labeled with both tritium and deuterium. 7-Ethylguanine was isolated from the nucleic acids of lung (not a target organ for DEN) and liver, but not from the intestine. The ethylation by DEN did not involve formation of diazoethane as intermediate. A small quantity of a second alkylated base, 7-methylguanine, in the rat liver RNA appeared to arise from a trace of methylethylnitrosamine in both the tritium-labeled DEN and the deuterium-labeled DEN.