Enzootic hepatitis A infection in cynomolgus monkeys (Macaca fascicularis).

During a toxicology study in cynomolgus (long-tailed or crab-eating) monkeys (Macaca fascicularis), a randomly distributed incidence of significantly increased hepatic enzyme activity was observed. Premedication hepatic enzyme activity in all monkeys of this study was normal, but increased alanine aminotransferase (ALT) activity was found in 4 of the 24 animals 2 weeks after initiation of the study and in 10 of 24 at 4 weeks. A drug-related effect was considered unlikely initially because the increases were not doserelated, and a 3-year review of 655 cynomolgus monkeys revealed a 15-20% incidence of increased hepatic enzyme activity. Good correlation was subsequently established between increased hepatic enzyme activity, active hepatitis A virus (HAV) infection, and histomorphologic confirmation of hepatitis (chronic periportal inflammation). Follow-up viral serodiagnostic screening of resident macaques revealed an overall incidence of anti-HAV IgG in 80% (155/193) of cynomolgus and in 70% (14/20) of rhesus monkeys. Serial screening demonstrated that several initially negative monkeys became seropositive for anti-HAV IgG, and a few acquired active infection (anti-HAV IgM). Among newly acquired cynomolgus monkeys, 2.5% (2/80) had an acute HAV infection, and 35% (28/80) eventually tested positive for anti-HAV IgG while quarantined in the primate facility. The characterization of an enzootic HAV infection in incoming monkeys posed a significant risk for the primate colony and handlers. Rigorous sanitation, isolation, and quarantine procedures, including personnel training and additional protective clothing for personnel working in the primate colony, reduced tho potential for transmission and arrested the outbreak. Experimenters should be cautious in ascribing toxicity to a test article based solely on increased hepatic enzyme activity associated with chronic periportal inflammation.

Tissue response in dogs to dense hydroxylapatite implantation in the femur.

Six and eight years after the implantation of both granular and solid ceramic hydroxylapatite forms in the femurs of beagle dogs, histologic examination demonstrated that the implants were totally encased in dense mature bone. The endosteal and periosteal surfaces appeared normal, and no resorption of the solid implants was observed. However, at six years, a few granules located at the periosteal surface showed interdigitation of connective tissue stalks, with large multinucleated cells at the interface with the implant. This phenomenon may represent some limited resorptive activity on the surfaces of these few isolated granules. Initially, radiographs showed exaggerated degrees of bone deposition on the endosteal surface under the solid implants (discs), as opposed to a less pronounced endosteal response to the implants of particulate material. In some cases, particularly with the disc implants, cracks were found in the ceramic material six years after implantation. These cracks, on staining, were found to be filled with amorphous material, suggesting an osseous matrix. The results of these long-term studies indicate that such hydroxylapatite implants in bone are highly biocompatible. Bone deposition and maturation on the implant surface resulted in a homogeneous bone/implant interface in which the host tissues appeared to respond to the implant as if it were normal bone.

Evaluation of hydroxylapatite root implants in baboons.

To determine if there is a need for close conformation of a hydroxylapatite implant to the root socket for successful prevention of alveolar ridge resorption after tooth extraction, second premolars and lateral incisors of six mature baboons were extracted bilaterally, and custom-fitted root replicas or half-length, plug-shaped forms of hydroxylapatite were implanted in the fresh sockets. Results showed that precise fit, as achieved by a custom-fitted implant, was not necessary for success. All implants were completely accepted, with evidence of new bone formation continuing for up to six months, the duration of the study. No resorption of the alveolar ridge was observed by gross or microscopic examination.

Nephrotoxic and ototoxic effects of hydroxygentamicin in cats.

Hydroxygentamicin, an aminoglycoside antibiotic, was administered subcutaneously to cats in doses up to 160 mg base/kg daily for 10 to 13 weeks. Gentamicin and a vehicle solution were tested as positive and negative control, respectively; in one test netilmicin was also included for comparative purposes. Several parameters, including serum urea nitrogen, serum creatinine, organ/body weight ratios, serum and tissue concentrations of the antibiotics, and renal pathology, were determined to ascertain the nephrotoxic potential of the three aminoglycosides. In addition, observations for the onset of ataxia and impairment of righting reflex were made during the course of the studies to compare the vestibular ototoxic effects of the three antibiotics. Although serum urea nitrogen and serum creatinine values increased markedly in those cats which eventually died or were sacrificed moribund, these parameters in survivors were slightly but not significantly higher than controls. Serum concentrations of the drugs were proportional to the doses administered, but renal concentrations were approximately two and five times as high for netilmicin and gentamicin, respectively, as they were for equivalent doses of hydroxygentamicin. The morphological changes observed in the kidney of cats given 60 mg base/kg of hydroxygentamicin were slightly less than those seen in cats administered 10 mg base/kg of gentamicin; similarly, kidney changes in cats given netilmicin were observed approximately twice as frequently as they were in those receiving equivalent doses of hydroxygentamicin. The nephrotoxic effects of aminoglycoside antibiotics were directly related to renal drug concentration and not to serum concentration, which was a function of dose.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparative nephrotoxicity of hydroxygentamicin and other aminoglycosides in rats.

The nephrotoxicity of hydroxygentamicin and amikacin was examined in young adult Fischer 344 rats. Serum creatinine (SCr) and urea nitrogen (BUN) levels were not significantly affected following sc injection of 80 or 160 mg/kg/day of hydroxygentamicin for 15 days. However, 250 mg/kg of amikacin produced significant increases in both parameters and in kidney/body weight ratios. The ratios were also significantly increased after 80 or 160 mg/kg of hydroxygentamicin, but kidneys of rats receiving amikacin were considerably heavier than those of rats treated with hydroxygentamicin. The antibacterial potency of 250 mg/kg of amikacin is comparable to that of 100 mg/kg of hydroxygentamicin. Additional studies, directly comparing hydroxygentamicin, a mutational biosynthetic, with gentamicin or netilmicin, all at 40, 80, and 160 mg base/kg, and incorporating renal function parameters as well as SCr, BUN, organ weight, tissue concentration, and kidney histopathology, revealed a characteristic pattern typical of aminoglycoside nephrotoxicity in mature adult male rats. In most parameters, values in rats given hydroxygentamicin or netilmicin were normal and comparable to those in controls, but kidney/body weight ratios were significantly increased at high doses. However, kidneys of rats medicated with gentamicin at comparable doses were considerably heavier than those of hydroxygentamicin-treated rats. Significant nephrotoxicity also was seen in rats given low doses of gentamicin or netilmicin. Eosinophilic granulation and vacuolization of renal proximal tubular epithelium, interstitial inflammation, and tubular dilation were observed microscopically with all three drugs in the following descending order of severity: gentamicin greater than netilmicin greater than hydroxygentamicin. The effects on proximal tubular epithelial cells following treatment with amikacin, netilmicin, or hydroxygentamicin correlated reasonably well with renal drug concentrations, but drug concentrations of gentamicin, which produced the most extensive kidney injury, were lower than those of the other three aminoglycosides. Elevated SCr or BUN were indicative of the presence of nephrosis, but early stages of tubular epithelial degeneration were not predicted by increases in BUN or SCr. Although minimal or mild nephrosis was seldom predicted by polyuria, proteinuria, or changes in osmolality, effects observed in renal function parameters usually correlated well with renal histopathology. However, a decrease in osmolality correlated best with enlarged kidneys and changes in renal morphology.

Histologic observation of soft tissue responses to implanted, multifaceted particles and discs of hydroxylapatite.

The effects on soft tissue of short- and long-term implants of ceramic hydroxylapatite (durapatite) are reported. In rats, after one week and one, six, and 12 months of subcutaneous implantation of hydroxylapatite in multifaceted particle and disc form, neither particles nor discs had resulted in any microscopically remarkable inflammation. In beagle dogs, after subcutaneous implantation of hydroxylapatite as multifaceted particles and as discs for seven and 24 days, nine months, and two and six years, no implant migration was observed. Encapsulation of particles and discs of increasing thickness was seen throughout the six years of observation. Except for a few isolated macrophages seen within the connective tissue stroma at seven and 24 days, no evidence of inflammation was found. In tissue sections taken at six months from beagle dogs in which multifaceted particles had been placed subperiosteally beneath the gingiva, dense connective tissue was observed adjacent to and surrounding the individual particles. These results show that hydroxylapatite implanted subcutaneously in rats and dogs produces little or no inflammatory response and is compatible with tissue irrespective of the shape of the implant.

Activity of quinfamide against natural infections of Entamoeba criceti in hamsters: a new potent agent for intestinal amoebiasis.

A novel tetrahydroquinolinyl ester, quinfamide, administered orally in multiple doses for 3 days had an ED50 of 0.25 mg/kg/day (total dose 0.75 mg/kg) for eradicating Entamoeba criceti in hamsters in several tests. It was significantly more active by direct comparison than 3 commercially available amoebicides and at least as active as 2 other esters of the parent compound, 1-(dichloroacety)-1,2,3,4-tetrahydro-6-quinolinol. After administration of a single dose, ED50 calculations for quinfamide averaged 0.9 mg/kg. Quinfamide was considerably more active than the other tetrahydroquinolinols, diloxanide furoate and teclozan, and it was approximately 1.5 times more active than etofamide; a statistical significance between the latter 2 drugs could be demonstrated in one of 4 tests. Administered prophylactically, quinfamide was shown to protect hamsters from re-infection with E. criceti. It also inhibited propagation of E. histolytica in vitro at a concentration of 20 microgram/ml. No adverse effects were noted in rodents after a single dose as high as 10 g/kg. Daily administration to monkeys of doses up to 500 mg/kg for as long as 37 days produced no pharmacological aberrations during or after medication; haematological studies and urine analyses were normal and no gross or microscopical tissue changes attributable to quinfamide were observed. No toxicity was revealed following acute (2 g/kg) and chronic (500 mg/kg/day x 31 days) administration of the drug to dogs and rats, respectively.

1-(Dichloroacetyl)-1,2,3,4-tetrahydro-6-quinolinol esters. New potent antiamebic agents.

A series of 1-(dichloroacetyl)-1,2,3,4-tetrahydro-6-quinolinols and certain O-acyl derivatives thereof have been prepared and shown to be potent antiamebic agents in the Entamoeba criceti infected hamster model. Compounds were compared with etichlordifene and diloxamide and one of them, 1-(dichloroacetyl)-6-(2-furoyloxy)-1,2,3,4-tetrahydroquinoline (4), was selected for human trial.

Chemotherapeutic activity of combination doses of chloroquine, pyrimethamine, and sulfamethoxy-diazine, a long-acting sulfanilamide, against Plasmodium berghei infections in mice.

During the course of antimalarial screening, it was discovered that sulfamethoxydiazine, a long-acting sulfanilamide extensively used in genitourinary tract infections, not only was effective against Plasmodium berghei infections in mice when administered alone but also was active when used in combination with chloroquine, in effect making it possible to use half as much of the latter drug as normally required to achieve the same results. The triple combination of chloroquine, sulfamethoxydiazine, and pyrimethamine, when administered in a ratio of 30:10:1, was found to be potentiating against both blood-induced and sporozoite-induced P. berghei NK(65) infections. Mean effective dose values were calculated for chloroquine, sulfamethoxydiazine, and pyrimethamine against blood-induced P. berghei infection, and when a combination of the three drugs was administered therapeutically in the ratio given above, only one-tenth as much chloroquine, one-thirtieth as much sulfamethoxydiazine, and one-sixtieth as much pyrimethamine were needed to cure 50% of murine infections as was needed for each drug alone. The triple combination also showed enhanced activity against lethal sporozoite-induced P. berghei NK(65) infection in A/J strain mice.