Department of Health and Human Services Public Health Service, Grant Application

Principal investigator/Program Director (Last, first, middle):

Duesber, Peter H.

Newell et al., 1985a; Haverkos and Dougherty, 1988b; Lesbian & Gay Substance Abuse Planning
Group, 1991a; Lesbian & Gay Substance Abuse Planning Group, 1991b; Duesberg, 1992b;
Duesberg, 1992a; Morgan et al., 1992).
Here we propose to examine the immunotoxic and sarcomagenic potential of nitrite inhalants -
perhaps the psychoactive drug most frequently used by AIDS risk groups - either as autonomous
pathogens or as cofactors of the retrovirus, HIV. Our proposal is based on the following toxicological
and epidemiological evidence.
b) Toxicity of nitrites. Because inhalation of alkyl nitrites relaxes smooth muscles, it has been
prescribed since 1867 against angina pectoris and heart pain at doses of 0.2 ml (Cox et al., 1983;
Newell et al., 1985a; Shorter, 1987; Seage et al., 1992). No AIDS-defining diseases have been
reported at these doses in patients with those relatively severe, terminal cardiovascular diseases (Cox et
al., 1983; Shorter, 1987), possibly because the patients did not inhale enough and did not live long
enough to develop them. However, the consumption of higher doses and for durations of up to 10
years have been reported to precede AIDS. Under these conditions, nitrites are used as sexual
stimulants in private homes, bath houses or as "room deodorizers" in gay discos (Newell et al., 1985a;
Lauritsen and Wilson, 1986; Dax et al., 1988; Rappoport, 1988; Adams, 1989).
Given a molecular weight of 120 daltons, 1 ml (or 0.01 mole) of amyl nitrite contains 6 x 1021
molecules, or 6 x 107 nitrite molecules for every one of the 1014 cells in the human body - clearly
enough for abundant toxicity. Alkylnitrites are directly toxic as they are rapidly hydrolyzed in vivo to
yield nitrite ions, which react with all biological macromolecules (Osterloh and Olson, 1986; Maikel,
1988). Indeed, immediate and late toxicities have been observed in recreational users who have
inhaled milliliters of nitrite inhalants (Newell et al., 1985a; Dax et al., 1988; Schwartz, 1988). Addicts
with 0.5 mM nitrite derivatives and 70% methemoglobin (nitrite-oxidized hemoglobin) in the blood
have been recorded (Osterloh and Olson, 1986). Toxicity for the immune system, the central nervous
system, the hematologic system and pulmonary organs has been observed after short exposure to
nitrites in humans and in animals (Newell et al., 1985a; Dax et al., 1988; Newell et al., 1988; Wood,
1988). In 1982, Goedert et al. found that the helper to suppresser T-cell ratio was lower in
homosexual men who had used volatile nitrite inhalants than among nonusers (Goedert et al., 1982).
Further, alkylnitrites were shown to be both mutagenic and potentially carcinogenic in animals(Jorgensen and Lawesson, 1982; Hersh et al., 1983; Newell et al., 1985a; Mirvish et al., 1988;
Newell et al., 1988).
By comparing the AIDS risk factors of homosexual men with AIDS to those without, Newell
et al. and others determined a direct "dose-response gradient": the higher the nitrite usage, the greater
the risk for AIDS (Marmor et al., 1982; Newell et al., 1985b; Haverkos and Dougherty, 1988b) and
deduced a 7-10 year lag time between chronic consumption of nitrites and Kaposi's sarcoma (Newell
et al., 1985a). In view of this, several investigators have proposed that nitrite inhalants cause
-pulmonary and skin Kaposi's sarcoma and pneumonia by direct toxicity on the skin, the oral mucosa
and on lymphocytes (Centers for Disease Control, 1982; Marmor et al., 1982; Haverkos et al., 1985;
Mathur-Wagh et al., 1985; Newell et al., 1985b; Lauritsen and Wilson, 1986; Mirvish and Haverkos,
1987; Haverkos, 1990; Morgan et al., 1992).
Because of their toxicity, a prescription requirement was instated for the sale of nitrite inhalants
by the rood and Drug Administration in 1969 (Newell et at., 1985a), and because of an "AIDS link"
(Cox, 1986) the sale of nitrites was banned by the U.S. Congress in 1988 (Public Law 100-690)
(Haverkos, 1990) and by the "Crime Control Act of 1990". Moreover the US. Food and Drug
Administration limits nitrites as food additives to less than 200 ppm, because of direct toxicity and
because "they have been implicated in an increased incidence of cancer" (Lewis, 1989).
c) Evidence for a nitrite use epidemic among homosexuals at risk for AJDS. An early CDC

study of 420 homosexual men attending clinics for sexually transmitted diseases in New York, Atlanta
and San Francisco reported that 86.4% had frequently used amyl- and butyl nitrites as sexual
PHS 398 (Rev.9/91) Page 25
Number pages consecutively at the bottom throughout the application. Do not use suffixes such as Sa, 3b.