Use of log-linear analysis to construct explanatory models for TDBP- and AFB1-induced mutation spectra in lacI transgenic animals

Mutation spectra recovered from lacI transgenic animals exposed in separate experiments to tris-(2,3-dibromopropyl)phosphate (TDBP) or aflatoxin B1 (AFB1) were examined using log-linear analysis. Log-linear analysis is a categorical procedure that analyses contingency table data. Expected contingency table cell counts are estimated by maximum likelihood as effects of main variables and variable interactions. Evaluation of hierarchical models of decreasing complexity indicates when significant explanatory power is lost by the sequential omission of interactions between variables. Use of this technique allows construction of the most parsimonious models to account for mutation spectra obtained in the two experiments. The resulting statistical models are consistent with previous analyses of these data and with biological explanations for causes of the observed spectra.     

Highly attenuated vaccine strains of simian immunodeficiency virus protect against vaginal challenge: inverse relationship of degree of protection with level of attenuation

Three different deletion mutants of simian immunodeficiency virus (SIV) that vary in their levels of attenuation were tested for the ability to protect against mucosal challenge with pathogenic SIV. Four female rhesus monkeys were vaccinated by intravenous inoculation with SIVmac239Delta3, four with SIVmac239Delta3X, and four with SIVmac239Delta4. These three vaccine strains exhibit increasing levels of attenuation: Delta3 < Delta3X 相似文献   

Urogenital system of the spotted hyena (Crocuta crocuta Erxleben): a functional histological study

The unique urogenital anatomy and histology of female spotted hyenas (Crocuta crocuta Erxleben) was reexamined to identify adaptations of "structure" that enable/facilitate urination, mating, and parturition through the clitoris. Unusual features of penile anatomy required for meeting ceremonies and successful mating through a clitoral point of insertion were also examined. As reported previously, the upper urogenital tract of the female spotted hyena is typical of other carnivores and consists of the oviducts, uterine horns, uterine body, and vagina. An anatomically defined cervix is absent, even though a histologically defined transition zone between the uterine body and vagina was demonstrated. Adaptive features of the upper genital tract were a helical-shaped uterine cavity, extensive smooth muscle in the uterus and vagina, and a newly discovered submucosal mucous urogenital gland (SMUG) located immediately caudal to the vagina. The extensive smooth muscle facilitates the expulsion of the large pups at parturition through the recurved birth canal. Secretions of the SMUG provide lubrication and protection for the urogenital mucosa during mating and parturition. Two types of "erections" are suggested by behavioral observations: the common hemodynamic erection required for insertion and thrusting by the male, and phallic "flipping" that commonly occurs earlier in the mating sequence and is sometimes seen during meeting ceremonies. Phallic "flipping" appears to be accomplished by the coordinated contractions of the large ischiocavernosus and retractor muscles acting on the semirigid organ. The extremely thick tunica albuginea and interstitial collagen of the common corporal body of the penis and clitoris gives the flaccid phallus some degree of rigidity even in the resting state in males and nulliparous females. Phallic "flipping" implies a hinge region in which flexibility is the key feature. Such a proximal hinge region of the male and female phallus was defined and was notable for its diminished collagen content. The urogenital sinus traversing the clitoris was specialized for distensibility, thus facilitating receipt of the penis during mating and for passage of the infant to the tip of the glans clitoris, where it emerges at parturition. The morphology of the glans penis is notable for the tapered common corporal body that extends to the distal tip of the glans. This adaptation is suggested to be required for a clitoral (as opposed to a vaginal) point of insertion during mating. Finally, additional segments of erectile tissue devoid of a thick collagenous capsule were demonstrated in the glans penis and glans clitoris, which appear to account for the "partially-locking" of the male into the female during the late stages of a mating sequence. Taken together, it is evident that the unusual sexual behaviors of the male and female spotted hyenas are facilitated by unique structural modifications of the relevant organs.     

COP9 signalosome components play a role in the mating pheromone response of S. cerevisiae

A family of genetically and structurally homologous complexes, the proteasome lid, Cop9 signalosome (CSN) and eukaryotic translation initiation factor 3, mediate different regulatory pathways. The CSN functions in numerous eukaryotes as a regulator of development and signaling, yet until now no evidence for a complex has been found in Saccharomyces cerevisiae. We identified a group of proteins, including a homolog of Csn5/Jab1 and four uncharacterized PCI components, that interact in a manner suggesting they form a complex analogous to the CSN in S. cerevisiae. These newly identified subunits play a role in adaptation to pheromone signaling. Deletants for individual subunits enhance pheromone response and increase mating efficiency. Overexpression of individual subunits or a human homolog mitigates sst2-induced pheromone sensitivity. Csi1, a novel CSN interactor, exhibits opposite phenotypes. Deletants also accumulate Cdc53/cullin in a Rub1-modified form; however, this role of the CSN appears to be distinct from that in the mating pathway.     

Induction of mucosal homing virus-specific CD8(+) T lymphocytes by attenuated simian immunodeficiency virus

Induction of virus-specific T-cell responses in mucosal as well as systemic compartments of the immune system is likely to be a critical feature of an effective AIDS vaccine. We investigated whether virus-specific CD8(+) lymphocytes induced in rhesus macaques by immunization with attenuated simian immunodeficiency virus (SIV), an approach that is highly effective in eliciting protection against mucosal challenge, express the mucosa-homing receptor alpha4beta7 and traffic to the intestinal mucosa. SIV-specific CD8(+) T cells expressing alpha4beta7 were detected in peripheral blood and intestine of macaques infected with attenuated SIV. In contrast, virus-specific T cells in blood of animals immunized cutaneously by a combined DNA-modified vaccinia virus Ankara regimen did not express alpha4beta7. These results demonstrate the selective induction of SIV-specific CD8(+) T lymphocytes expressing alpha4beta7 by a vaccine approach that replicates in mucosal tissue and suggest that induction of virus-specific lymphocytes that are able to home to mucosal sites may be an important characteristic of a successful AIDS vaccine.     

Selenium and immunocompetence in patients with head and neck cancer

This randomized double-blind placebo-controlled study aimed to determine whether oral intake of 200 μg/d of sodium selenite, a dose within the safe and adequate daily intake (50–200 μg/d) recommended by the U.S. Food and Nutrition Board, will abrogate depressed or enhance normal-level immune functions of patients receiving therapy for squamous cell carcinoma of the head and neck. Subjects were given one selenium/placebo tablet/d for 8 wk, beginning on the day of their first treatment for the disease (e.g., surgery, radiation, or surgery and radiation) and their immune functions were monitored. Supplementation with selenium (Se) during therapy resulted in a significantly enhanced cell-mediated immune responsiveness, as reflected in the ability of the patient’s lymphocytes to respond to stimulation with mitogen, to generate cytotoxic lymphocytes, and to destroy tumor cells. The enhanced responsiveness was evident during therapy and following conclusion of therapy. In contrast, patients in the placebo arm of the study showed a decline in immune responsiveness during therapy, which was followed, in some patients, by an enhancement, but the responses of the group remained significantly lower than baseline values. The data also show that at baseline, patients entered in the study had significantly lower plasma Se levels than healthy individuals, and patients in stage I or II of disease had significantly higher plasma selenium levels than patients in stage III or IV of disease.     

Mutant frequencies and mutation spectra of dimethylnitrosamine (DMN) at the lacI and cII loci in the livers of Big Blue transgenic mice

The lacI gene in Big Blue transgenic rodents has traditionally been used as a surrogate gene for in vivo mutations. Recently, a more efficient and less expensive assay involving direct selection in the smaller lambda cII gene has been developed. Little is known, however, about the comparative sensitivity of the two loci or their influence on the recovered mutation spectrum following mutagen treatment. We have compared the mutation frequency (MF) and mutational spectrum (MS) of lacI and cII from the same DNA samples isolated from the liver of control and dimethylnitrosamine (DMN)-treated mice. A three-fold (p<0.01) increase in the MF was observed at both loci in the DMN-treated group compared to the corresponding control groups. While the DMN-induced mutation spectrum at lacI was significantly different from its corresponding spontaneous mutation spectrum (p<0.001), the mutation spectrum at cII (p>0.28) was not. The mutation spectra at the two loci from the DMN-treated mice resembled each other but the 4, 2.5 and 12-fold increase in the mutation frequency of A:T>T:A transversions, single base deletions and deletions of more than four base pairs, respectively, at lacI, altered the spectra significantly (p<0.007). The number of mutations of these classes at cII was also increased, but the fractions were lower than at lacI. The spontaneous mutation spectra at the cII and lacI loci resembled each other except for the seven-fold increase in G:C相似文献   

Subchronic administration of phenobarbital alters the mutation spectrum of lacI in the livers of Big Blue transgenic mice

Phenobarbital (PHE) is a liver carcinogen in B6C3F1 mice and a weak mutagen that does not appear to form DNA adducts. To investigate PHE mutagenicity in vivo, B6C3F1 Big Blue(R) male transgenic mice harboring the lambdaLIZ shuttle vector containing the lacI target gene were fed PHE at 2500 ppm for 180 days. A modest increase in the mutant frequency (MF) from 5.02+/-2.4x10(-5) in the control group to 6.88+/-0.754x10(-5) in the PHE-treated group, which was marginally different (p<0.05), was obtained. To better assess the relevance of this increase in MF, a random collection of mutants from each PHE-exposed mouse was sequenced. After correcting for clonal expansion, which is the most conservative approach, the MF in the PHE-treated mice decreased to 6.39+/-1.02x10(-5), an insignificant difference (p=0.10) from that in control group. Despite this modest increase in MF, the mutation spectrum obtained from the PHE-exposed group was significantly different (pA:T transitions remained the same in the two spectra. It is postulated that the increase in transversions at G:C base pairs found in the PHE-derived spectrum is likely due to oxidative damage as a result of induction of CYP2B isozymes by the chronic administration of PHE. Results from this study demonstrate that PHE alters the spectrum of mutations, rather than inducing a significant global increase in the MF. The PHE-derived spectrum of lacI mutants from the liver of Big Blue(R) B6C3F1 male mice was remarkably similar (p=0.8) to that generated by oxazepam (OX), a compound which also induces CYP2B isozymes following chronic administration of the drug.     

Low shear stress preferentially enhances IKK activity through selective sources of ROS for persistent activation of NF-kappaB in endothelial cells

NF-B signaling pathway has been known to play a major role in the pathological process of atherogenesis. Unlike high shear stress, in which the NF-B activity is transient, our earlier studies have demonstrated a persistent activation of NF-B in response to low shear stress in human aortic endothelial cells. These findings partially explained why low shear regions that exist at bifurcations of arteries are prone to atherosclerosis, unlike the relatively atheroprotective high shear regions. In the present study, we further investigated 1) the role of NF-B signaling kinases (IKK and ) that may be responsible for the sustained activation of NF-B in low shear stress and 2) the regulation of these kinases by reactive oxygen species (ROS). Our results demonstrate that not only is a significant proportion of low shear-induced-kinase activity is contributed by IKK, but it is also persistently induced for a prolonged time frame. The IKK activity (both and ) is blocked by apocynin (400 µM), a specific NADPH oxidase inhibitor, and diphenyleneiodonium chloride (DPI; 10 µM), an inhibitor of flavin-containing oxidases like NADPH oxidases. Determination of ROS also demonstrated an increased generation in low shear stress that could be blocked by DPI. These results suggest that the source of ROS generation in endothelial cells in response to low shear stress is NADPH oxidase. The DPI-inhibitable component of ROS is the primary regulator of specific upstream kinases that determine the persistent NF-B activation selectively in low shear-induced endothelial cells. upstream B kinases; laminar shear stress; oxidative stress; atherogenesis; reactive oxygen species