The pharmacokinetics and metabolism of diclofenac in chimeric humanized and murinized FRG mice.

The pharmacokinetics of diclofenac were investigated following single oral doses of 10 mg/kg to chimeric liver humanized and murinized FRG and C57BL/6 mice. In addition, the metabolism and excretion were investigated in chimeric liver humanized and murinized FRG mice. Diclofenac reached maximum blood concentrations of 2.43 ± 0.9 µg/mL (n = 3) at 0.25 h post-dose with an AUCinf of 3.67 µg h/mL and an effective half-life of 0.86 h (n = 2). In the murinized animals, maximum blood concentrations were determined as 3.86 ± 2.31 µg/mL at 0.25 h post-dose with an AUCinf of 4.94 ± 2.93 µg h/mL and a half-life of 0.52 ± 0.03 h (n = 3). In C57BL/6J mice, mean peak blood concentrations of 2.31 ± 0.53 µg/mL were seen 0.25 h post-dose with a mean AUCinf of 2.10 ± 0.49 µg h/mL and a half-life of 0.51 ± 0.49 h (n = 3). Analysis of blood indicated only trace quantities of drug-related material in chimeric humanized and murinized FRG mice. Metabolic profiling of urine, bile and faecal extracts revealed a complex pattern of metabolites for both humanized and murinized animals with, in addition to unchanged parent drug, a variety of hydroxylated and conjugated metabolites detected. The profiles in humanized mice were different to those of both murinized and wild-type animals, e.g., a higher proportion of the dose was detected in the form of acyl glucuronide metabolites and much reduced amounts as taurine conjugates. Comparison of the metabolic profiles obtained from the present study with previously published data from C57BL/6J mice and humans revealed a greater, though not complete, match between chimeric humanized mice and humans, such that the liver humanized FRG model may represent a model for assessing the biotransformation of such compounds in humans.

The pharmacokinetics and metabolism of lumiracoxib in chimeric humanized and murinized FRG mice.

The pharmacokinetics and metabolism of lumiracoxib were studied, after administration of single 10mg/kg oral doses to chimeric liver-humanized and murinized FRG mice. In the chimeric humanized mice, lumiracoxib reached peak observed concentrations in the blood of 1.10±0.08µg/mL at 0.25-0.5h post-dose with an AUCinf of 1.74±0.52µgh/mL and an effective half-life for the drug of 1.42±0.72h (n=3). In the case of the murinized animals peak observed concentrations in the blood were determined as 1.15±0.08µg/mL at 0.25h post-dose with an AUCinf of 1.94±0.22µgh/mL and an effective half-life of 1.28±0.02h (n=3). Analysis of blood indicated only the presence of unchanged lumiracoxib. Metabolic profiling of urine, bile and faecal extracts revealed a complex pattern of metabolites for both humanized and murinized animals with, in addition to unchanged parent drug, a variety of hydroxylated and conjugated metabolites detected. The profiles obtained in humanized mice were different compared to murinized animals with e.g., a higher proportion of the dose detected in the form of acyl glucuronide metabolites and much reduced amounts of taurine conjugates. Comparison of the metabolic profiles obtained from the present study with previously published data from C57bl/6J mice and humans, revealed a greater though not complete match between chimeric humanized mice and humans, such that the liver-humanized FRG model may represent a useful approach to assessing the biotransformation of such compounds in humans.

Extensive double humanization of both liver and hematopoiesis in FRGN mice.

Preclinical research in animals often fails to adequately predict the outcomes observed in human patients. Chimeric animals bearing individual human tissues have been developed to provide improved models of human-specific cellular processes. Mice transplanted with human hematopoietic stem cells can be used to study human immune responses, infections of blood cells and processes of hematopoiesis. Animals with humanized livers are useful for modeling hepatotropic infections as well as drug metabolism and hepatotoxicity. However, many pathophysiologic processes involve both the liver and the hematolymphoid system. Examples include hepatitis C/HIV co-infection, immune mediated liver diseases, liver injuries with inflammation such as steatohepatitis and alcoholic liver disease. We developed a robust protocol enabling the concurrent double-humanization of mice with mature hepatocytes and human blood. Immune-deficient, fumarylacetoacetate hydrolase (Fah(-/-)), Rag2(-/-) and Il2rg(-/-) deficient animals on the NOD-strain background (FRGN) were simultaneously co-transplanted with adult human hepatocytes and hematopoietic stem cells after busulfan and Ad:uPA pre-conditioning. Four months after transplantation the average human liver repopulation exceeded 80% and hematopoietic chimerism also was high (40-80% in bone marrow). Importantly, human macrophages (Kupffer cells) were present in the chimeric livers. Double-chimeric FRGN mice will serve as a new model for disease processes that involve interactions between hepatocytes and hematolymphoid cells.

Fibronectin levels of unstimulated saliva from naval recruits with and without chronic inflammatory periodontal disease.

Past studies have suggested that gingival crevicular fluid is produced more readily in persons with severe periodontal diseases than in persons with healthy gingivae. In this study, salivary fibronectin was selected as an index of total gingival crevicular fluid flow. Our purpose was to determine whether a relationship could be found between salivary fibronectin level and periodontal disease status. Unstimulated saliva was collected from 20 healthy and 20 periodontally-diseased naval recruits. The periodontally-diseased subjects included 10 with localized juvenile periodontitis and 10 with moderate to severe periodontitis. Mean subject ages and salivary flow rates were similar for the 2 groups. Although 2 of the periodontally-diseased subjects showed unusually high fibronectin levels, the mean level for the remaining 18 subjects did not differ significantly from the mean of the healthy group, and no association of periodontal disease status with salivary fibronectin content was seen. Consequently, it was not evident from salivary fibronectin levels that the content of gingival crevicular fluid in unstimulated whole saliva differed significantly for persons with or without severe periodontal disease, except possibly for extreme cases of disease.

Quantitative relationship of Treponema denticola to severity of periodontal disease.

The Treponema denticola content of plaque was quantitatively estimated for samples taken from periodontitis patients as well as periodontally healthy subjects among two separate human populations. The populations studied included military volunteers and civilians at a university dental clinic. The plaque samples from each population were grouped according to pocket depth measurements at the collection site. A biotin-avidin enzyme-linked immunosorbent assay procedure was developed with a monoclonal antibody specific for a serovariety of T. denticola. T. denticola was present at significantly elevated levels in plaque samples collected from deep-pocket sites of patients with severe periodontitis relative to the healthy controls and a group with moderate disease. The ratio of T. denticola content per milligram of plaque in the deep pocket groups to that of the other two groups was about 2:1 for both populations. This is the first quantitative evidence of a positive relationship between a specific spirochete species and severe periodontitis.

Diagnostic value of a coagglutination procedure using monoclonal antibodies to Bacteroides gingivalis.

A specific monoclonal antibody against Bacteroides gingivalis was bound to particles coated with protein A and evaluated for use in a coagglutination test. B. gingivalis was the only organism tested which gave a specific positive reaction with the CoA reagent. Subgingival plaque samples were collected from 217 patients diagnosed as having periodontitis. Organisms that gave biochemical reactions which indicated they were B. gingivalis were isolated from eleven of the 217 gingival pockets. These eleven strains were the only organisms which gave a positive reaction using the CoA test.

Radiographic evaluation of juvenile periodontitis (periodontosis).

Of 49,380 male naval recruits who were screened for juvenile periodontitis (JP), 270 were clinically diagnosed as having the disease. Full-mouth radiographs identified 182 of these 270 patients as having JP with extensive bone loss on permanent first molars and/or incisors. These 182 patients, 137 (75.3%) of whom were black, were further classified into Type I: bone loss involving first molars and/or incisors and up to two additional teeth; Type II: involvement of first molars/incisors and several additional teeth; and Type III: generalized involvement (more than 14 teeth) but with bone loss notably more extensive on the first molars and/or incisors. Of the 182 patients, 129 (70.9%) were Type I; 43 (23.6%) were Type II, and 10 (5.5%) were Type III. The molars were involved more frequently than the incisors; more than one molar was always involved, with or without incisor involvement. Most cases had minimal or no radiographic caries, and 46% had demonstrable calculus. Of the remaining 88 cases from the 270 with the initial diagnosis of JP, 63 demonstrated severe bone loss on more than 14 teeth, with many of the teeth being involved to the same degree. These cases were termed rapidly progressive periodontitis. Six of the 88 cases had bone loss on only one tooth surface of the dentition. These cases were termed acute localized destruction of alveolar bone. The status of the other 19 cases could not be determined.