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Proliferol is an investigational new drug containing lidocaine hydrochloride 0.25%, dextrose 12.5%, glycerin 12.5%, and phe­nol 1.0% in aqueous solution. Despite extensive human experience with similar drugs administered by intraligamentous injection for chronic musculoskeletal disorders, little is known concerning preclinical toxicity. The purpose of this study was to assess the acute toxicity of intramuscular Proliferol in 96 (48 male, 48 female) Charles River strain rats, which were randomly assigned to low-(1×), medium- (5×), or high- (10×) dose Proliferol (derived from a human dose of 20 ml on a volume per bodyweight basis), or high-dose saline placebo. Observations included clinical observations, biochemistry, hematology, urinalysis, and full histopathology after 24 h or 14 days. There were no signs of ill health or reaction to treat­ment, and gait and body temperature were within normal limits. Biochemistry findings at 24 h included elevated aspartate aminotransferase, alanine aminotransferase, and haptoglobin; at 14 days all values were within normal ranges. Urinalysis findings at 24 h included increased urobilinogen and blood in all dose groups com­pared with placebo. Urine concentrations of phenol and lidocaine were greatest at 2 h and absent at 24 h. Histopathology findings included localized acute inflammatory soft tissue changes at the injection sites at 24 h and skeletal muscle regeneration at 14 days, which were consistent with the anticipated mechanism of action of Proliferol. There was no evidence of systemic toxicity from intra­muscular injection of Proliferol in rats at up to 10× the human dose.

Background. There is a need to prevent or minimize bone loss associated with antiretroviral treatment (ART) initiation. We compared maraviroc (MVC)- to tenofovir disoproxil fumarate (TDF)–containing ART. Methods. This was a double-blind, placebo-controlled trial. ART-naive subjects with human immunodeficiency virus type 1 RNA load (viral load [VL]) >1000 copies/mL and R5 tropism were randomized to MVC 150 mg or TDF 300 mg once daily (1:1), stratified by VL

This article presents views of a discipline termed safety pharmacology or general pharmacology. This is an area that provides information through empirical studies on new pharmacologic agents at doses above those thought to be efficacious and the no-toxicologic-effect level (NOEL) above which unwanted effects might occur. The usefulness of batteries of tests is discussed, and comments are made about the value of these in the drug developmental process.

Prolotherapy is one of the many treatments available for chronic musculoskeletal disorders. A commonly used drug contains dextrose 12.5%, glycerin 12.5%, phenol 1.0%, and lidocaine hydrochloride 0.25% in aqueous solution (recently termed Proliferol). For chronic low back pain, this is injected into lumbosacral ligaments to stimulate connective tissue repair. Despite generally positive clinical results, the toxicity of this drug is not well characterized and was assessed in 48 (24 male, 24 female) Yucatan miniature swine randomly assigned to low (1×), medium (5×), or high (10×) dose or saline placebo. Outcomes included clinical observations, clinical chemistry, hematology, coagulation, urinalysis, toxicokinetics, and full gross and microscopic histopathology after 24 hours or 14 days. Findings attributable to Proliferol after 24 hours included dose-response elevations in alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and creatine kinase, which returned to normal after 14 days. There were no remarkable findings in hematology, coagulation, or urinalysis. Urine concentrations of lidocaine and phenol both peaked after 8 hours. Histopathology findings after 24 hours included hemorrhage, inflammation, necrosis, and vascular changes in the ligaments and adjacent soft tissues at the sites of injection. After 14 days, there was evidence of repair under way, with fibrosis and skeletal muscle regeneration at the injection sites.

Influenza/pneumonia is the only infections disease(s) among the top ten causes of death in the U.S. (Check, 1984). Influenza virus kills an average of over 20,000 people each year in the United States (Katz, 1985). In addition, the illness has a substantial economic impact. Schoenbaum (1987) estimated that the total direct cost of influenza (health care cost) exceeds $1 billion per year in the U.S., with the actual cost (including lost productivity) on the order of $3 to $5 billion per year. Yet, until recently at least, only about 20% of the over 43 million Americans at risk of death from influenza by virtue of their age or underlying medical condition receive vaccinations (Mostow, 1986; Check, 1984). If there is a close match between the vaccine strain and the circulating strain, protection can be 70 to 80% in the adult population (Douglas, 1990). Protection may be lower in the infirm elderly who are at greatest risk (Gross et al, 1989).

Monochloroacetone was introduced in 1914 as a war gas, and presently it has a number of uses as a chemical intermediate. Apart from its biological properties as a lacrimator and vesicant, it is not a well-studied compound toxicologically. A series of acute toxicity studies were done using different routes of administration. An Ames mutagenicity test also was performed. These data were compared to manufacturing use information and data published in the literature. It is recommended that direct contact with liquid and vapor be prevented through strict engineering controls and that air concentrations be kept below 1 ppm as a ceiling concentration.

Methyl fluorosulfonate (Magic Methyl®), an active met hy la ting agent used by research chemists, was studied for acute oral toxicity, acute inhalation toxicity, ocular irritation, and dermal irritation. This compound is very hazardous and may have been responsible for at least one human death. The results of these studies confirmed that Magic Methyl is markedly toxic by all routes studied and particularly by the inhalation route since the LC50 value for rats was found to be between 5 and 6 ppm.

Ronidazole was evaluated for mutagenic potential using in vitro microbial tests and in vivo studies in mice. The microbial test used the histidine requiring mutants of Salmonella typhimurium with and without a rat liver microsomal activation system (Ames test). The studies in mice included the dominant lethal test, micronucleus test and cytogenetic assays. Ronidazole was given orally in doses of 50, 100 and 200 mg/kg/day in the in vivo studies. In the dominant lethal test, groups of male mice were treated for five consecutive days before being mated with untreated females. In the micronucleus test, the mice were administered the compound for 2 or 5 consecutive days; they were killed 6 h after the last dose and bone marrow was examined for the presence of micronuclei in developing erythrocytes. In the cytogenetic assays, bone marrow cells in metaphase were examined for chromosome aberrations, 6, 24 and 48 h after mice were treated acutely with the test compound. In addition, similar examinations of chromosomes were made on mice given five consecutive dosages of ronidazole and killed 6 h after the last dose. The results of the various in vivo studies did not suggest that ronidazole would be mutagenic for the mammal. Ronidazole at concentrations of 10 and 50 mug/plate was found to increase the number of back mutations of missense mutants in the in vitro bacterial test. This finding confirms the results of Voogd et al. [19]. Incorporation of the microsomal activation system had no effect on the mutagenic capability of the test compound. In conclusion, although ronidazole was shown to be mutagenic in in vitro bacterial systems, the in vivo systems did not suggest that the compound would be mutagenic for the mammal.

There are many possible micronucleus assays involving different test organisms and tissues. Because micronuclei arise from chromosomal fragments or chromosomes that are not incorporated into daughter nuclei at the time of cell division, the assay detects both clastogens and agents that affect the spindle apparatus. We know of no case in which micronuclei and chromosomal breakage (or loss) have been shown to occur independently of one another in any dividing cell population. This relationship is so close that false-positives and false-negatives (insofar as the detection of tissue-specific chromosome damage is concerned) should be determined primarily by the statistics of sampling. The production of micronuclei in various experimental organisms has been reviewed. Although there are several promising experimental approaches such as the use of meiotic plant cells or human cells in culture, only one form of the assay, the in vivo mammalian bone-marrow polychromatic erythrocyte (PCE) assay, has been sufficiently developed to be considered a standard assay. More than 150 chemicals have been tested in this assay, with varying degrees of rigor. The data from the literature have been summarized and evaluated in light of the work Group's recommendations for an adequate test. The standards for an adequate test are an important part of the recommendations. These standards, although based on the most recent information available to us, are subject to change because this assay is still evolving. The most important recommendations in this report are: (1) at least 500 PCE should be examined from each of 8 animals to detect an increase of about 4‰ (per thousand) PCE when the background is less than 4 per 1000, (2) sampling should be extended to at least 72 h after the initial treatment, with sampling intervals no greater than 24 h, and (3) the highest possible doses should be used. The success rate of the assay to detect chemicals designated by the Environmental Protection Agency (EPA) as carcinogens is difficult to estimate for several reasons. First, few chemicals designated as noncarcinogens were studied, although in routine testing noncarcinogens are expected to be much more common than carcinogens. Hence, the rate of false-positives (insofar as the detection of cancer is concerned), which ought to be one of the strongest features of the assays, could not be estimated. Second, few chemicals have been tested as rigorously as this report recommends. Hence, the rate of false-negatives is almost certainly overestimated. (It is, nevertheless, obvious that false-negatives are to be expected for any tissue-specific in vivo assay like the micronucleus assay. For example diethylnitrosamine, which produces chromosomal aberrations, micronuclei, and cancer in the liver, is not detected in the bone-marrow micronucleus assay.) Third, many carcinogens are species-specific and this fact has not been taken into account. Considering these caveats, the uncorrected detection rate of the chemicals designated as carcinogens by EPA is about 50%. We believe that this would have been significantly higher had all tests been performed according to the test criteria. Further improvements in the assay are to be expected and these may lead to improvements in its success rate. Recent developments are discussed.

Hydrocarbon activation and mutation induction in the L5178Y mouse lymphoma mutation assay were studied as a function of the concentration of an exogenously added rat liver homogenate activation system (S20). Four polycyclic aromatic hydrocarbons were tested at constant dosage levels with 4 concentrations of S20 for ability to induce forward mutations at the thymidine kinase (TK) locus. With all 4 hydrocarbons, increasing S20 concentration resulted in a significant (p less than 0.001) decrease in mutation frequency and corresponding increase in survival. In the absence of S20, no significant mutation induction was observed. Within 5 min after the addition of S20 to a reaction mixture containing [3H]benzo[a]pyrene [B(a)P] (3 micrograms/ml) and appropriate cofactors, 70% of the B(a)P was metabolized with a high S20 concentration, whereas only 14% was metabolized with a low S20 concentration. In the absence of S20, the cells did not metabolize B(a)P. Incubation of B(a)P with a low concentration of S20 in the presence of calf thymus DNA (1.5 mg/ml) resulted in twice as much covalently bound B(a)P per mg DNA (33 pmoles/mg) as incubation with a high concentration of S20 (15 pmoles/mg). The amount of the major B(a)P-DNA adduct, (+/-)-7 beta,8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene-deoxyguanosine, per mg DNA was 5 times greater with low S20 concentration than with high (11.6 pmoles/mg and 2.1 pmoles/mg, respectively). These results suggest that the decrease in mutation frequency with high S20 concentrations results from the rapid oxidation of the hydrocarbon which reduces the opportunity for mutagenic derivatives that are formed from B(a)P to interact with DNA and induce mutation.

The use of the electron microscope is becoming more widespread in laboratories throughout the world, including those serving the various disciplines of environmental health. As a result, practicing industrial hygienists may have opportunities to use the instrument. This paper points out some of the areas in which the electron microscope has been employed in the solution of problems in the field of occupational health.

The potential cytogenetic effect of benzene was studied by means of the micronucleus test in developing erythrocytes of male and female mice at dosages ranging from 0.0625 to 2.0 ml/kg/day. Groups of mice were given two daily oral doses and sacrificed 6, 18, 24 or 48 h, or 5, 9 or 16 days after the last dose. The doses were given 24 h apart. Bone marrow from the mice was processed for visualization of micronuclei in polychromatic erythrocytes. The results showed a significant (P < 0.05) increases in the numbers of polychromatic erythrocytes with micronuclei from mice sacrificed 6 h after being given dosages of 0.25 ml/kg/day or more of benzene. Similarly, significant (P < 0.05) increases in polychromatic erythrocytes with micronuclei were seen in mice given 0.125 ml/kg/day or more of benzene and sacrificed 18 or 24 h after the second dose. However, in a second experiment negative results were obtained at 24 h from mice at 0.0625, 0.125 and 0.25 ml/kg. One group of mice (0.25 ml/kg/day and killed at 48 h) showed significant increases in the numbers of polychromatic erythrocytes with micronuclei. Mice killed 5 days after treatment with 2 doses of 0.125 ml/kg or more of benzene showed significant numbers of micronuclei in polychromatic erythrocytes. The groups of mice at 0.5, 1.0 or 2.0 ml/kg/day and killed 9 or 16 days after the second dose of benzene showed values that were comparable to the corresponding negative controls. In all cases there were sufficient numbers of cells to analyze. Although benzene is a known hematopoietic poison and bone-marrow depressant, there were sufficient numbers of cells to evaluate in the micronucleus test. Thus, these experiments uphold the usefulness of the micronucleus test as a screening procedure in cytogenetics.

A series of novel benzoheterocyclic [(methoxyphenyl)amino]oxoalkanoic acid esters has been prepared. These compounds were tested as inhibitors of rat polymorphonuclear leukocyte 5-lipoxgenase (LO) in vitro and as inhibitors of leukotriene D4 (LTD4) and ovalbumin (OA) induced bronchospasm in the guinea pig (GP) in vivo. In general, inhibitory activity against 5-LO, LTD4, and OA was broadest for benzthiazole-containing analogues (benzthiazole greater than benzimidazole much greater than benzoxazole, benzofuran). The most potent 5-LO inhibitor, 4-[[3-(2-benzthiazolylmethoxy)-phenyl]hydroxyamino]-4-oxobutanoic acid methyl ester (7), had an IC50 of 0.36 microM. Compound 7, however, was inactive vs. OA. The most potent compound in vivo, 4-[[3-[(1-methyl-2-benzimidazolyl)methoxy]phenyl]-amino] -4-oxobutanoic acid methyl ester 4, inhibited both LTD4- and OA-induced bronchospasm by 83% and 60%, respectively, at 50 mg/kg intraduodenally. Compound 4 was studied in the Ames assay employing five strains of bacteria (TA1535, TA1537, TA1538, TA98, and TA100) with and without S-9 rat liver enzyme metabolic activation, and there was no significant number of reversions noted.

Three nitroparaffins (nitroethane, 1-nitropropane, and 2-nitropropane) were studied in the Salmonella typhimurium/mammalian microsome (Ames) test, with and without microsomal activation systems. Nitroethane and 2-nitropropane also were studied in an in vivo mutagenic (micronucleus) test. These studies were undertaken because these solvents are widely used in the chemical and pharmaceutical industries and 2-nitropropane was reported to cause liver cancer in rats exposed by the inhalation route. Neither nitroethane nor 1-nitropropane was active in the Ames test with Salmonella tester-strains TA1537, TA92, TA98, or TA100. However, 2-nitropropane produced a significant increase in revertants in all of these tester strains, particularly strain TA100, where 3 microliter/plate doubled the number of revertants in the presence of microsomal enzymes. Negative results were obtained with both nitroethane and 2-nitropropane in micronucleus tests. These studies have shown that 2-nitropropane has the potential for causing point mutations in a microbial test system. However, this compound probably will not cause a chromosome mutation of the clastogenic type.

Cyclization of the anilines (I) and subsequent etherification produce the nitrophenolates (V) which are transformed into the amides (VI).

Ganglioneuromas (GNs) are benign, slow-growing, rare soft-tissue tumors that arise from the sympathetic nervous system and comprise less than 1% of all soft-tissue neoplasms. Although GNs are slow-growing, they can and will invade bone and pressure local adjacent structures by their continued growth. In a 20-year-old woman, GN produced a large presacral mass. This case illustrates the diagnostic features at various stages of growth, histopathology, and treatment of GN.