Incidence and potentiation of external and internal fetal anomalies resulting from chlordiazepoxide and amitriptyline alone and in combination.

The teratogenic potential of a combination of chlordiazepoxide (Cdz) and amitriptyline (Amt) was examined with regard to both internal and external anomalies. Timed pregnant golden hamsters were given a single intraperitoneal injection on day 8 of gestation of one of the following: chlordiazepoxide hydrochloride (28.5 mg/kg), amitriptyline hydrochloride (70.3 mg/kg), Cdz-Amt combination (28.5 mg/kg Cdz + 70.3 mg/kg Amt, in order to retain the 1:2.5 dose ratio utilized in a clinically-used preparation of these agents), or saline vehicle (control). Fetuses were recovered on gestation day 15 following maternal sacrifice. Cranial malformations were analyzed in Bouin's-fixed fetuses by making 1-mm coronal sections through each head, whereas visceral anomalies were examined following general dissection of each body. Amt alone produced a significant (P less than 0.05) incidence of bent tail and encephalocele, whereas Cdz significantly (P less than 0.05) altered the male:female ratio of surviving fetuses when compared with saline-injected controls. The Cdz-Amt combination caused significant increases in cranial malformations, open eye, bent tail, abnormal lung, and urogenital anomalies. The teratogenic effects of potentiation between the components of this combination are discussed in terms of external and internal malformations.

Comparative teratogenicity of chlordiazepoxide, amitriptyline, and a combination of the two compounds in the fetal hamster.

A combination of chlordiazepoxide and amitriptyline maternally administered as a single intraperitoneal injection on day 8 of gestation in the fetal hamster produced predominantly central nervous system anomalies including exencephaly and encephalocoele. In addition, omphalocoele, spinal flexion, and microcephaly were noted. A dose response relationship was found in which a maternal dose range of 13/33 mg/kg--33/83 mg/kg chlordiazepoxide/amitriptyline produced 7-92 percent fetal anomalies. Combination drug dose levels up to 23/58 mg/kg produced no maternal mortality. However, higher levels did result in a marked dose dependent mortality rate. The teratogenic potential of the combined drugs is much more pronounced than that of either drug administered alone since chlordiazepoxide at a maternal dose range of 280/3100 mg/kg produced 3-55 percent fetal anomalies, and amitriptyline at a maternal dose range of 60-100 mg/kg produced 6-45 percent fetal anomalies. The majority of these aberrant fetal developmental entities also were classifiable as exencephaly and encephalocoeles. Dose-dependent maternal mortality was observed at all dose levels for each drug administered separately.

Haloperidol teratogenicity in the fetal hamster.

Haloperidol, administered as a single intraperitoneal injection on day 8 of gestation, produced a variety of fetal malformations in hamsters. Fetal defects included exencephaly, cranioschisis, microphthalmia, anophthalmia, lower body hypodevelopment, and total body hypodevelopment. A dose-response relationship was evident as 80-245 mg/kg produced from 3 to 70% fetal anomalies.

Duration of interferon inhibition following single and multiple injections of morphine.

The increase in mouse serum interferon that occurs in response to poly I:C administration is markedly attenuated by injection of a narcotic preceding the polynucleotide challenge. A single injection of morphine is capable of inhibiting the mechanism(s) responsible for increasing serum interferon levels for a period of at least 9 days following the narcotic injection. Daily single injections of morphine for 5 or 10 days prior to poly I:C challenge do not amplify the extent of the decrease in serum interferon over that measured for a single injection, even though the amount of narcotic injected is increased on each successive day of the 5 or 10 day period. The implications of this relationship for the phenomenon of narcotic tolerance are evaluated and discussed as they relate to the immune processes.

Role of spleen in the interferon-lowering action of morphine.

1. Polyinosinie-polycytidylic acid (Poly I:C) administered intraperitioneally to mice produced an increase in the level of interferon in the tissues of the spleen, liver, lung, and kidney. The level of circulating serum interferon also was increased by the polynucleotide. 2. The highest titer of interferon, presumably resulting from the greatest response to poly I:C was measured in the splenic tissue. 3. Morphine inhibited the poly I:C induced increase in all tissues and circulating serum. The decrease in interferon titer was similar in spleen and serum. 4. These results may indicate the serum level of interferon was more closely related and dependent upon events occuring in the spleen than in the other tissues.

Action of naloxone on the interferon-lowering activity of morphine in the mouse.

The serum titer of interferon produced by polyinosinic:polycytidylic acid (poly I:C) was studied in mice following subcutaneous injections of either morphine alone, naloxone alone, morphine and naloxone together, morphine preceded by naloxone, naloxone preceded by morphine, or a sequence of injections of naloxone, morphine, and naloxone again. Poly I:C injected without the other drugs present served as the control. Both morphine and naloxone produced a significant decrease in the ability of the mice to increase the serum titer of interferon in response to poly I:C. Naloxone appeared to function as an antagonist to morphine when administered following the morphine injection, but not when injected before or simultaneously with the opiate. When naloxone was injected both before and following the morphine injection, results were interpreted to indicate the possibility that naloxone could function as both an agonist and antagonist under this condition.

Congenital malformations of the central nervous system produced by narcotic analgesics in the hamster.

Maternal dose--fetal teratogenic response data were obtained for a variety of narcotic and related compounds by single subcutaneous injections of the drugs into pregnant hamsters during the critical periods of central nervous system organogenesis. The number of abnormal fetuses from females injected with diacetylmorphine (heroin), thebaine, phenazocine, pentazocine, propoxyphene, and methadone increased as the maternal dose of the compounds was increased. By contrast, morphine, hydromorphone, and meperidine produced an increase in the number (per cent) of fetal anomalies only up to a certain maternal dose level. Further increases in maternal dose levels did not produce additional fetal anomalies. Comparative studies of single and multiple maternal doses indicated that diacetylmorphine (heroin) and methadone produced a four- to sixfold increase in fetal anomalies with repetitive doses whereas the percentage of malformed fetuses remained the same with hydromorphone (Dilaudid). The narcotic antagonists nalorphine, naloxone, levallophan, and cyclazocine blocked the teratogenic effects of both single and multiple doses of the narcotics.

Effect of morphine, hydromorphone, methadone, mescaline, trypan blue, vitamin A, sodium salicylate, and caffeine on the serum interferon level in response to viral infection.

The ability of morphine, hydromorphone, methadone, mescaline, trypan blue, vitamin A, sodium salicylate and caffeine to alter the circulating serum level of interferon was investigated in mice. Sodium salicylate, and caffeine increased the serum level of interferon; however, the interferon increase produced by sodium salicylate was dose-dependent, i.e. low doses increased interferon titers, high doses decreased the titers. All other compounds decreased the circulating level of interferon. These results suggest that compounds of varied structure and biological activity, and possessing teratogenic potential, are capable of depressing the circulating level of interferon. Some of these "anti-interferon" compounds, i.e. morphine, hydromorphone, methadone, depressed interferon levels at all doses whereas sodium salicylate decreased the interferon level only at relatively high dose levels.

Effect of ascorbic acid, sodium salicylate, and caffeine on the serum interferon level in response to viral infection.

The ability of ascorbic acid, sodium salicylate, and caffeine to alter the circulating serum level of interferon was investigated in mice. The animals were singly injected subcutaneously with one of the compounds, 4-8 h later again singly injected intraperitoneally with poly I:C, and bled 6-8 h afterward. The sera from the mice were assayed for interferon titer by the use of the plaque inhibition method utilizing vesicular stomatitis virus. Ascorbic acid, sodium salicylate, and caffeine increased the serum level of interferon; however, the increase produced by sodium salicylate was dose-dependent, i.e. low doses increased interferon titers, high doses decreased the titers. Caffeine produced minimal increases in the interferon titer. These observations suggest that a potential prophylactic result may occur in virus infections from administration of the three compounds either singly or in combination at the proper concentration.

Congenital malformations induced by mescaline, lysergic acid diethylamide, and bromolysergic acid in the hamster.

Malformations of the brain, spinal cord, liver, and other viscera; body edema; and localized hemorrhages were found in fetal hamsters from mothers injected subcutaneously with a single dose of mescaline, lysergic acid diethylamide, or 2-bromo-D lysergic acid diethylamide on the 8th day of pregnancy. In addition, all three drugs produced an increase in the percentages of small fetuses per litter, of resorptions, and of fetal mortality.