Interstrain Differences Research Articles

Flurbiprofen Pharmacokinetics: Interstrain and Sex Differences in Rats

Interstrain differences in the pharmacokinetic behaviour of drugs can influence their plasma concentration-time profiles and the extent of drug exposure, thereby influencing the outcome of animal toxicity studies. The pharmacokinetics of flurbiprofen was investigated in Wistar and Sprague–Dawley rats, the two most commonly used strains in toxicity studies. Flurbiprofen was administered orally at a dose of 2 mg kg−1 in a solution of polyethylene glycol) 400, to male and female Wistar and Sprague–Dawley rats. Serial blood samples were collected at 0, 0.25, 0.5, 1, 2, 4, 6, 8 and 12h postdose by jugular vein cannulation. Wistar rats exhibited significantly higher peak plasma concentrations with a significant decrease in Vd/F. Within each strain, animals of each sex had similar absorption and distribution kinetics. However, male rats in both strains exhibited faster elimination and clearance, with a corresponding decrease in AUC0-∞. These differences were significant only in the Wistar strain. The results confirm the occurrence of significant interstrain and sex differences in the disposition of flurbiprofen in Wistar and Sprague–Dawley rats.

Multiplex PCR Assay for the Detection of Five Putative Virulence Genes Encoded in Verotoxigenic Escherichia coli Plasmids

The aim was to perform a pentavalent PCR assay for the detection of putative virulence genes encoded in VTEC plasmids, katP, espP, subA, stcE, and ehxA. The five-specific primer pairs used in the assay do not interfere with each other and generate amplification products of 914, 774, 556, 399, and 262bp. It was selected at random 39 strains belonged to 20 serotypes in order to evaluate the multiplex in a wide variety of strains. The results of this study indicate that it is possible to perform simultaneous amplification and search for recognized plasmid-encoded virulence markers from different E. coli serotypes and apply this technique to the genetic characterization of E. coli strains isolated from reservoirs, foods or patients. This complementary technique is a useful tool to detect interstrain differences for epidemiological studies and to provide information that could be related to the risk of human infection.

Comparative Gene Expression Analysis in the Skeletal Muscles of Dysferlin-deficient SJL/J and A/J Mice

Quantitative real-time polymerase chain reaction (qRT-PCR) analysis was conducted to determine whether or not there are interstrain or site-dependent differences in the gene expression profiles of skeletal muscles in SJL/J and A/J mice as dysferlinopathy models. Upon analysis by qRT-PCR, SJL/J mice showed a trend of increased gene expression level of uncoupling protein 2 in the rectus femoris and longissimus lumborum at 30 weeks of age when dystrophic lesions became histopathologically pronounced. Heme oxygenase 1 and S100 calcium binding protein A4 were upregulated in the rectus femoris, longissimus lumborum and abdominal muscles, in which dystrophic lesions occur more commonly in SJL mice. The gene expression levels of heat shock protein 70 in most muscles of A/J mice were lower than those of BALB/c mice as control. SJL/J mice exhibited a marked lowering of decay-accelerating factor 1/CD55 gene expression level in all studied muscles except for the heart at all ages compared with that of BALB/c mice. This study showed that there were some interstrain differences in the gene expres sion profiles of skeletal muscles between SJL/J and A/J mice. Further investigation is required to reveal whether these alterations of the expression levels are the cause of dystrophic changes or occur subsequent to muscle damage.

Interstrain Differences in the Liver Effects of Trichloroethylene in a Multistrain Panel of Inbred Mice

Interstrain Differences in the Liver Effects of Trichloroethylene in a Multistrain Panel of Inbred Mice

Interstrain differences in susceptibility to non-alcoholic steatohepatitis

Interstrain differences in susceptibility to non-alcoholic steatohepatitis

Interstrain differences in the liver effects of trichloroethylene in a multistrain panel of inbred mice.

Trichloroethylene (TCE) is a widely used industrial chemical and a common environmental contaminant. It is a well-known carcinogen in rodents and a probable carcinogen in humans. Studies utilizing panels of mouse inbred strains afford a unique opportunity to understand both metabolic and genetic basis for differences in responses to TCE. We tested the hypothesis that strain- and liver-specific toxic effects of TCE are genetically controlled and that the mechanisms of toxicity and susceptibility can be uncovered by exploring responses to TCE using a diverse panel of inbred mouse strains. TCE (2100 mg/kg) or corn oil vehicle was administered by gavage to 6- to 8-week-old male mice of 15 mouse strains. Serum and liver were collected at 2, 8, and 24 h postdosing and were analyzed for TCE metabolites, hepatocellular injury, and gene expression of liver. TCE metabolism, as evident from the levels of individual oxidative and conjugative metabolites, varied considerably between strains. TCE treatment-specific effect on the liver transcriptome was strongly dependent on genetic background. Peroxisome proliferator-activated receptor-mediated molecular networks, consisting of the metabolism genes known to be induced by TCE, represent some of the most pronounced molecular effects of TCE treatment in mouse liver that are dependent on genetic background. Conversely, cell death, liver necrosis, and immune-mediated response pathways, which are altered by TCE treatment in liver, are largely genetic background independent. These studies provide better understanding of the mechanisms of TCE-induced toxicity anchored on metabolism and genotype-phenotype correlations that may define susceptibility or resistance.

The Accessory Genome of Pseudomonas aeruginosa

Pseudomonas aeruginosa strains exhibit significant variability in pathogenicity and ecological flexibility. Such interstrain differences reflect the dynamic nature of the P. aeruginosa genome, which is composed of a relatively invariable "core genome" and a highly variable "accessory genome." Here we review the major classes of genetic elements comprising the P. aeruginosa accessory genome and highlight emerging themes in the acquisition and functional importance of these elements. Although the precise phenotypes endowed by the majority of the P. aeruginosa accessory genome have yet to be determined, rapid progress is being made, and a clearer understanding of the role of the P. aeruginosa accessory genome in ecology and infection is emerging.

Interstrain Differences of In Vitro Metabolic Stability and Impact on Early Drug Discovery

With the extensive use of different strains of mice and rats in in vivo efficacy models, lack of relevant metabolic clearance data among strains has been a concern. Metabolic clearance is an important parameter impacting drug discovery, and it is often used as a compound selection filter. Metabolically stable compounds are often preferred, and will have a better chance to achieve the desired exposure in vivo. The present study examined strain differences in mouse and rat, using 96 compounds which spanned a wide range of intrinsic clearances. The in vitro clearances were determined using liver microsomes from commonly used strains of mouse (BALB/c, C57BL/6J, and CD-1) and of rat (Sprague-Dawley, Fischer, and Wistar Han). There were few discrepancies in the interpretation of the in vitro intrinsic clearance results within species for the 96 compounds tested in mouse and rat liver microsomes. This data gives us confidence that the phase I hepatic clearance can be determined using only one strain of mouse or rat liver microsomes.

Genetic Susceptibility to the Delayed Sequelae of Neonatal Respiratory Syncytial Virus Infection Is MHC Dependent

Respiratory syncytial virus (RSV) is a major cause of respiratory morbidity, resulting in hospitalization for bronchiolitis in some infected infants that is associated with wheeze in later life. Genetic factors are known to affect the severity of the sequelae after RSV infection, but the complexity of the temporal and genetic effects makes it difficult to analyze this response in studies in man. Therefore, we developed a murine genetic model to analyze the sequelae occurring after RSV infection in early life. Haplotype-based genetic analysis of interstrain differences in severity identified the MHC as an important genetic determinant. This was confirmed by analysis of responses in congenic mice with different MHC haplotypes. We also found that susceptible strains had high CD8 levels during secondary infection. Analysis of first filial generation, second filial generation, and back-cross progeny produced by intercrossing resistant (H-2(k), C3H/HeN) and sensitive (H-2(b), BALB/c) strains indicated that susceptibility to sequelae after RSV infection was dominantly inherited but also segregated in a non-MHC-dependent manner. Thus, MHC haplotype and its effect on CD8 cell response is an important determinant of the outcome of neonatal RSV infection.

The Role of Interleukin-1 in Wound Biology. Part I

Wound healing is a multistep, complex process that involves the coordinated action of multiple cell types. Conflicting results have been obtained when conventional methods have been used to study wound biology. Therefore, we analyzed the wound response in a mouse genetic model. We analyzed inflammatory mediators produced within incisional wounds induced in 16 inbred mouse strains. Computational haplotype-based genetic analysis of inter-strain differences in the level of production of 2 chemokines in wounds was performed. An in vitro experimental analysis system was developed to investigate whether interleukin (IL)-1 could affect chemokine production by 2 different types of cells that are present within wounds. The level of 2 chemokines, keratinocyte-derived chemokine (KC) and macrophage inflammatory protein 1α, exhibited very large (75- and 463-fold, respectively) interstrain differences within wound tissue across this inbred strain panel. Genetic variation within Nalp1, an inflammasome component that regulates IL-1 production, correlated with the interstrain differences in KC and macrophage inhibitory protein 1α production. Consistent with the genetic correlation, IL-1β was shown to stimulate KC production by murine keratinocyte and fibroblast cell lines in vitro. Genetic variation within Nalp1 could contribute to interstrain differences in wound chemokine production by altering the amount of IL-1 produced.

The Role of Interleukin-1 in Wound Biology. Part II

In the accompanying paper, we demonstrate that genetic variation within Nalp1 could contribute to interstrain differences in wound chemokine production through altering the amount of interleukin (IL)-1 produced. We further investigate the role of IL-1 in incisional wound biology and its effect on wound chemokine production in vivo and whether this mechanism could be active in human subjects. A well-characterized murine model of incisional wounding was used to assess the in vivo role of IL-1 in wound biology. The amount of 7 different cytokines/chemokines produced within an experimentally induced skin incision on a mouse paw and the nociceptive response was analyzed in mice treated with an IL-1 inhibitor. We also investigated whether human IL-1β or IL-1α stimulated the production of chemokines by primary human keratinocytes in vitro, and whether there was a correlation between IL-1β and chemokine levels in 2 experimental human wound paradigms. Administration of an IL-1 receptor antagonist to mice decreased the nociceptive response to an incisional wound, and reduced the production of multiple inflammatory mediators, including keratinocyte-derived chemokine (KC) and macrophage inhibitory protein (MIP)-1α, within the wounds. IL-1α and IL-1β stimulated IL-8 and GRO-α (human homologues of murine keratinocyte-derived chemokine) production by primary human keratinocytes in vitro. IL-1β levels were highly correlated with IL-8 in human surgical wounds, and at cutaneous sites of human ultraviolet B-induced sunburn injury. IL-1 plays a major role in regulating inflammatory mediator production in wounds through a novel mechanism; by stimulating the production of multiple cytokines and chemokines, it impacts clinically important aspects of wound biology. These data suggest that administration of an IL-1 receptor antagonist within the perioperative period could decrease postsurgical wound pain.

Fermentative characteristics and fibrolytic activities of anaerobic gut fungi isolated from wild and domestic ruminants

Fermentative characteristics and fibrolytic enzyme activities of anaerobic gut fungi from wild (17 isolates) and domestic ruminants (15 isolates) were examined. In a medium containing 0.5% wheat straw and 0.02% cellobiose as energy source, activities of carboxymethyl cellulase (CMCase), avicelase, xylanase, acetyl esterase and protease produced by the fungal isolates were investigated. Average activity of CMCase (17.4 vs. 8.25 mIU ml−1), acetyl esterase (134 vs. 57 mIU ml−1) and protease (4400 vs. 1683 mIU ml−1) were significantly higher in isolates from wild ruminants than those from domestic ruminants. Xylanase and avicelase activities were comparable. When compared irrespective of source, fungal isolates having monocentric growth pattern produced more fibrolytic enzymes than isolates having polycentric growth pattern. CMCase, xylanase, avicelase activities were highest in Neocallimastix isolates. Acetyl esterase activity was highest in Piromyces and Neocallimastix isolates. Protease activity was highest in Piromyces isolates followed closely by Neocallimastix isolates. Between isolates from wild and domestic ruminants few differences were observed in pattern of carbohydrate utilisation and end products of fermentation. Inter-strain differences in the end product formation were apparent. All of the isolates produced acetate, lactate and formate; only a few isolates produced succinate. For isolation of superior fibrolytic isolates of anaerobic fungi, greater emphasis should be given to the screening of enzyme activities of isolates of genera Neocallimastix and Piromyces.

The post-antifungal effect (PAFE) of amphotericin B, nystatin, ketoconazole and 5-fluorocytosine and its impact on the colonization traits ofCandida glabrata

The post-antifungal effect (PAFE) has been shown to affect Candida pathogenicity, but there is little information on either PAFE or its association with the colonization traits of Candida glabrata. The objective of this study was to determine, in vitro, the PAFE on 14 C. glabrata isolates following exposure to amphotericin B (AMB), nystatin (NYS), ketoconazole (KETO) and 5-fluorocytosine (5FC). In addition, we evaluated the impact of PAFE on yeast adherence to buccal epithelial cells (BEC), cell-surface-hydrophobicity (CSH) and biofilm growth (BG) on denture acrylic surfaces. PAFE was induced following a 1-h exposure of yeasts to (x1-x4MIC) of AMB, NYS, KETO and 5FC in RPMI medium and, measured using automated turbidometry. The BEC adhesion, CSH and BG assays were performed by the methods of Kimura & Pearsall, Sweet et al., and Jin et al., respectively. Significant differences in PAFE (P < 0.001) were observed after exposure to AMB and NYS, but not KETO and 5FC. Following exposure to AMB, NYS, KETO and 5FC, significant inter-strain differences (P < 0.001) were observed in percentage terms in adhesion (39.0%, 43.48%, 38.28%, 35.07%) and biofilm growth (42.86%, 39.86%, 42.81%, 36.38%), respectively. Short exposure of C. glabrata to sub-cidal concentrations of antifungals modulates yeast growth and also affects some of their colonization traits.

Comparisons of water activity and temperature impacts on growth of Fusarium langsethiae strains from northern Europe on oat-based media

This study has examined the effect of water activity (a w, 0.995–0.90) and temperature (10–37 °C) on the lag phases prior to growth, growth rates and used models to develop two dimensional profiles for optimum and marginal conditions for two strains of Fusarium langsethiae from four northern European countries (UK, Norway, Sweden, and Finland) on an oat-based medium. Results showed that the optimum a w for growth was at 0.98–0.995 and 25 °C. The limit for growth of the strains was at 0.92–0.93 a w with minima of 10 °C. No growth occurred at 37 °C. The lag phases prior to growth were lowest under optimum conditions and extended to > 10 days at marginal conditions. Statistical analyses of intra and inter-strain differences in terms of both lag phases prior to growth and growth rates were not statistically significant. However, a w and temperature were statistically significant factors. Two dimensional profiles for strains from each country of origin were built to identify optimum and marginal conditions for F. langsethiae for the first time. These environmental profiles will be beneficial for improving the ecological knowledge of this species which is able to produce trichothecene mycotoxins in a range of temperate cereals.

Interstrain differences in the resistance to the infection with densonucleosis virus (Yamanashi isolate) in the silkworm, Bombyx mori

Interstrain differences in the resistance to the infection with densonucleosis virus (Yamanashi isolate) in the silkworm, Bombyx mori

Liver expression of the nuclear hormone receptors PPAR, LXR, and RXR and blood parameters of lipid and carbohydrate metabolism in strains of mice susceptible and resistant to hepatocarcinogenesis

Earlier it was shown that male mice of the DD/He strain were highly susceptible to ortho-aminoa-zotoluene (OAT) induced hepatocarcinogenesis, and resistant to spontaneous liver tumor development as compared to male mice of the CC57BR/Mv strain. In the present work we have made a comparative investigation of peroxisome proliferator-activated receptor (PPAR), liver X-receptor (LXR) and retinoic X-receptor (RXR) mRNA levels in liver as well as concentrations of corticosterone, glucose, lipids and insulin in blood of male DD/He and CC57BR/Mv mice. Using the multiplex RT-PCR method it was found that PPAR-α, PPAR-γ, RXR-α, and RXR-β mRNA content was significantly lower in the liver of DD mice than in CC57BR mice. No significant interstrain differences were found in the content of LXR-α and LXR-β mRNA. In DD mice there was more than the 3-fold decrease of blood content of corticosterone (involved in PPAR and RXR regulation). DD mice demonstrated a significant decrease in blood serum glucose and insulin concentrations as well as higher reactivity to insulin as compared with NN57BR mice. Elevated blood total cholesterol and HDL cholesterol levels were found in DD mice whereas triglyceride content was basically the same in both mouse strains. It is known that glucocorticoids, PPAR and RXR play crucial role in transcription regulation of the inflammation response. Therefore our data on decreased corticosterone level in blood, PPAR and RXR mRNA content in the liver of the DD suggest that sensitivity of this strain of mice to the development of OAT-induced hepatocarcinomas may be associated with increased inflammatory reaction to this carcinogen.

Respiratory pattern in awake rats: Effects of motor activity and of alerting stimuli

Our aim was to assess the impact of motor activity and of arousing stimuli on respiratory rate in the awake rats. The study was performed in male adult Sprague–Dawley (SD, n = 5) and Hooded Wistar (HW, n = 5) rats instrumented for ECG telemetry. Respiratory rate was recorded using whole-body plethysmograph, with a piezoelectric sensor attached for the simultaneous assessment of motor activity. All motor activity was found to be associated with an immediate increase in respiratory rate that remained elevated for the whole duration of movement; this was reflected by: i) bimodal distribution of respiratory intervals (modes for slow peak: 336 ± 19 and 532 ± 80 ms for HW and SD, p < 0.05; modes for fast peak 128 ± 6 and 132 ± 7 ms for HW and SD, NS); and ii) a tight correlation between total movement time and total time of tachypnoea, with an R 2 ranging 0.96–0.99 ( n = 10, p < 0001). The extent of motor-related tachypnoea was significantly correlated with the intensity of associated movement. Mild alerting stimuli produced stereotyped tachypnoeic responses, without affecting heart rate: tapping the chamber raised respiratory rate from 117 ± 7 to 430 ± 15 cpm; sudden side move — from 134 ± 13 to 487 ± 16 cpm, and turning on lights — from 136 ± 12 to 507 ± 14 cpm ( n = 10; p < 0.01 for all; no inter-strain differences). We conclude that: i) sniffing is an integral part of the generalized arousal response and does not depend on the modality of sensory stimuli; ii) tachypnoea is a sensitive index of arousal; and iii) respiratory rate is tightly correlated with motor activity.

Substrain Differences Reveal Novel Disease-Modifying Gene Candidates That Alter the Clinical Course of a Rodent Model of Multiple Sclerosis

Experimental autoimmune encephalomyelitis (EAE) is a rodent model of multiple sclerosis that is executed in animals by immunization with myelin Ag in adjuvant. The SJL/J autoimmune-prone strain of mouse has been used to model relapsing-remitting multiple sclerosis. However, significant variations in peak scores, timing of onset, and incidence are observed among laboratories, with the postacute (relapse) phase of the disease exhibiting significant inconsistency. We characterized two substrains of SJL/J mice that exhibit profoundly different EAE disease parameters. Induction of EAE in the first SJL/J substrain resulted in many cases of chronic EAE that was dominated by an aggressive B cell response to the immunizing Ag and to endogenous CNS Ags. In contrast, the other SJL/J substrain exhibited a relapsing-remitting form of EAE concomitant with an elevated number of cytokine-producing CD4(+) T cells in the CNS. Exploiting these interstrain differences, we performed a genome-wide copy number analysis on the two disparate SJL/J substrains and discovered numerous gene-dosage differences. In particular, one inflammation-associated gene, Naip1, was present at a higher copy number in the SJL/J substrain that exhibited relapsing-remitting EAE. These results demonstrate that substrain differences, perhaps at the level of genomic copy number, can account for variability in the postacute phase of EAE and may drive chronic versus relapsing disease.

Inter-Strain Differences of Serotonergic Inhibitory Pain Control in Inbred Mice

BackgroundDescending inhibitory pain control contributes to the endogenous defense against chronic pain and involves noradrenergic and serotonergic systems. The clinical efficacy of antidepressants suggests that serotonin may be particularly relevant for neuropathic pain conditions. Serotonergic signaling is regulated by synthesis, metabolisms, reuptake and receptors.ResultsTo address the complexity, we used inbred mouse strains, C57BL/6J, 129 Sv, DBA/2J and Balb/c, which differ in brain serotonin levels. Serotonin analysis after nerve injury revealed inter-strain differences in the adaptation of descending serotonergic fibers. Upregulation of spinal cord and midbrain serotonin was apparent only in 129 Sv mice and was associated with attenuated nerve injury evoked hyperalgesia and allodynia in this strain. The increase of dorsal horn serotonin was blocked by hemisectioning of descending fibers but not by rhizotomy of primary afferents indicating a midbrain source. Para-chlorophenylalanine-mediated serotonin depletion in spinal cord and midbrain intensified pain hypersensitivity in the nerve injury model. In contrast, chronic inflammation of the hindpaw did not evoke equivalent changes in serotonin levels in the spinal cord and midbrain and nociceptive thresholds dropped in a parallel manner in all strains.ConclusionThe results suggest that chronic nerve injury evoked hypernociception may be contributed by genetic differences of descending serotonergic inhibitory control.

Individual, age and sex differences in fiber type composition of slow and fast muscles of adult Lewis rats: comparison with other rat strains

We analyzed fiber type composition of soleus and extensor digitorum longus (EDL) muscles of 3- to 19-month-old male and female inbred Lewis rats using histochemical demonstration of mATPase activity. The rats were divided into four groups of the mean age of 3, 6, 9 and 14 months. We found that the soleus muscle of 3-month-old rats contained significantly more of fast 2A fibers and less of slow type 1 fibers compared to older rats, while no significant difference was found between female and male rats at any age group. In contrast, we found no significant difference in the EDL fiber type composition among the age groups, but we found that the EDL muscle of female rats contained significantly less 2A fibers and more 2B fibers than that of male animals. Our results thus revealed an age difference in the soleus muscle and a sex difference in the EDL muscle among postnatal Lewis rats. The number of slow type 1 fibers in the soleus muscle varied between 87 and 100% and that of 2A fibers between 13 and 0%. In the EDL the percentage of type 1 fibers varied between 2.6 and 8.7%, that of 2A fibers between 12.6 and 25.8% and that of 2B fibers between 70.4 and 81.6%. Both muscles thus exhibited a considerable degree of variability among individual animals even in the same age group. Furthermore, a comparison of the Lewis rats with literature data of other rat strains showed that the number of fast 2A fibers in the soleus muscle of 4-month-old and older animals decreased in this order: SHR>Lister Hooded>Fisher 344>Sprague-Dawley>Wistar>WBN/Kob>Lewis strain, being almost 20% in the SHR and less than 2% in the Lewis rats. In contrast, the "fastest" composition (judged according to the percentage of the fastest 2B fibers) of the EDL muscle was demonstrated by Lewis, Wistar and Fisher 344 rats (about 75%), while Sprague-Dawley and WBN/Kob rats contained only about 50% of 2B fibers. The percentage of slow type 1 fibers in the EDL was low in all strains (about 5%). Our results thus show that the individual, age and sex as well as inter-strain differences in muscle fiber type composition should not be ignored when comparing results of different studies. We also demonstrated that the inbred Lewis strain appears to have more "specialized" muscle composition, as its soleus is the "slowest" and its EDL is the "fastest" among the routinely used rat strains.