Narrow Results

It is not only important for maintenance and improvement of health, but also indispensable for diagnosis and remedy of diseases to make inquiries into the biological defence mechanisms. Yonezawa et al. came across an induction of yet unknown defence mechanism(s) in mice which acquired radioresistance two weeks after low dose X-irradiation with 0.5 Gy. The 30-day survival rate after midlethal X-irradiation of the mice significantly increased by the pre-irradiation, but contrary to the common knowledge on radiation protection, recovery of the blood cell counts of thrombocytes, leukocytes and erythrocytes were not stimulated by the pre-irradiation. This study was planned to find some keys to elucidate the mechanism for the acquired radioresistance. Metal ions are well known to be important for enzyme activities as well as for metabolisms. Eleven elements, Cl, K, Ca, Cr, Mn, Fe, Ni, Cu, Zn, Se and Br were analysed in mice sera by PIXE.

The objective of this study is to investigate the significance of natural killer (NK) cells and interleukin-2 in uterus in the early embryo loss (or resorption), and to elucidate the immunological modulation of maternal-fetal interface with Chinese herbal medicine Radix Scutellariae (Huang Qin) and Rhizoma Atractylodis (Bai Zhu). Lipopolysaccharide (LPS) was given via the tail vein to induce abortion in mice at day 7 of gestation. Uterine NK cells and IL-2 contents were analyzed by immunohistochemistry and enzyme linked immunosorbent assay (ELISA), respectively. The number of NK cells was found to be much higher (mean = 180 ± 39) in the decidua of LPS-treated abortion mice. But when the Chinese herbal medicine was used to prevent LPS-induced abortion, less NK cells (mean = 11 ± 4) were counted (p < 0.01). The mean value of IL-2 in LPS-treated mice was 5.25 ± 2.5938 pg/mg protein, higher than (p < 0.05) that of the herb prevention group, which was only 1.86 ± 0.9789 pg/mg protein. The results therefore indicate that the increase of NK cells in the decidua and IL-2 contents in the uterus in LPS-treated mice is closely related to the embryo loss, and that the Chinese herbal medicine prescription composed of Radix scutellariae and Rhizoma atractylodis has an anti-bortive effect through inhibition of maternal-fetal interface immunity.

The effects of chronic treatment with hot water extract of Bitter Melon (Momordica charantia L.) or Ginger Rhizome (Zingiber offifinale Rosc) on spontaneous mammary tumorigenesis were examined in SHN virgin mice. In mice given free access to extract of Bitter Melon (0.5%) or Ginger (0.125%) in drinking water, the development of mammary tumors was significantly inhibited. Furthermore, the former inhibited uterine adenomyosis with a common pathological background to mammary tumors and the latter inhibited mammary tumor growth. While the mechanism of the effects of these natural products remains to be clarified, there were no adverse effects of chronic treatment with these agents as estimated from body weight, food and water intake and various plasma component levels as well as external appearance. Thus, these natural products, popular in Japan as foodstuffs, also appear to have a health benefit.

We have found that the administration of a diet containing 5% hydroxyapatite (HAP) derived from pig and cattle bones, and exposure to far-infrared rays (FIR) markedly inhibited spontaneous mammary tumorigenesis in SHN mice. Thus, the effect of combined treatment with HAP and FIR on mammary tumorigenesis was examined. The significant inhibition of tumor development by individual treatment with HAP or FIR was not enhanced by combined treatment; instead, the decrease in the inhibitory effect of HAP with age was ameliorated. Associated with this, life span was elongated and a decline in ovarian function was prevented by HAP plus FIR. Normal and preneoplastic growth of mammary glands and plasma component levels were not significantly affected by any treatment. The findings indicate that HAP and FIR have characteristics common to most natural products; in combination with other agents, they have little additive effect, when each is highly active.

It has been generally accepted that Hwangryunjihwang-tang (H-tang) is a useful prescription for treating polydipsia and to prevent obesity induced by a high-fat diet. The aim of this study was to clarify whether H-tang improved reproductive dysfunction caused by obesity in mice. Mice were fed a high density protein and lipid diet for 4 weeks, followed by administration of H-tang at 480 mg/kg body weight per day for 4 days. Thereafter, changes of body weight, ovulation rate, in vitro and in vivo fertilization, embryonic development and implantation rate were measured. H-tang markedly reduced the body weight of obese mice fed a high-fat diet, but not mice fed a normal diet. H-tang significantly improved ovulation rates, in vitro and in vivo fertilization rates and embryonic development. These results indicate pharmacological reversal of reproductive dysfunction caused by obesity, perhaps by adjusting internal secretions and metabolic functions.

A traditional Chinese Medicine (TCM) formula, Guo Min Kang (GMK), has been used in clinics in China for allergic diseases, including type I immediate hypersensitivity, a potentially fatal disease, but its modulatory mechanism remains elusive. The aim of this study was to investigate the modulatory mechanisms of GMK in a mouse model of Ag-induced anaphylaxis. Ag (conalbumin) sensitized mice were treated with either PBS (sham) or GMK before (schedule A) or during (schedule B) sensitization, and various anaphylactic parameters were measured following Ag challenge, including symptom score, cutaneous hypersensitivity response, mast cell degranulation, plasma histamine levels and the levels of specific IgE and T-cell responses. Systemic anaphylaxis was investigated in mice immediately following Ag challenge, and the results showed that GMK-treated mice from both treatment schedules A and B showed significantly reduced symptom scores when compared with the sham-treated group. The reduction in symptom score was associated with a significant reduction in the level of Ag-induced cutaneous immediate hypersensitivity. Also, GMK was able to suppress Ag-induced IgE production and T-cell responses, while it spares mitogen (Con A)-induced T-cell response. Further, treatment of mice with GMK abrogated the levels of Ag-induced histamine release and significantly reduced the number of degranulated mast cells. No effect of GMK was observed on the levels of total IgE and plasma histamine in naive mice. These results provide a basis for the modulation effect of GMK and suggest a potential utility of GMK as a prophylactic and therapeutic agent.

Toki-shakuyaku-san (TSS), an herbal formula based on traditional Chinese medicine, is commonly used in obstetrics. To examine the effects of TSS on the normal mouse fetus and placenta, TSS was administered to normal pregnant mice and their placentas and fetuses were studied. First, the effects of maternal TSS treatment on implantation were investigated. Administration of TSS from gestation day 0.5 (G0.5) to G6.5 showed that litter size was not altered at embryonic day 11.5 (E11.5), but the number of resorbed fetuses was slightly decreased. Then, to investigate effects on fetal and placental growths after implantation, TSS was administered from G5.5. At E14.5, the body weight of fetuses from TSS-treated dams was significantly increased. Gene expression of insulin-like growth factor 2 (Igf2), one of the most important modulators of fetal growth, was significantly increased in the placentas and fetuses of TSS-treated dams. In addition, the expression of particular placental developmental genes and nutrient transporter genes was significantly increased in TSS-treated placentas. At E18.5, after longer-term administration of TSS, fetal and placental weights were not altered, but the expression of the placental developmental and nutrient transporter genes remained elevated compared with controls. These results suggest that maternal TSS treatment in normal mice enhances the expression of Igf2, placental developmental genes and nutrient transporter genes, resulting in increased fetal weight. No obvious changes were observed in the expression of these genes after longer-term maternal TSS treatment.

Hypoxia-inducible factor-1 (HIF-1) is an $\alpha ∕\beta$ dimeric transcription factor. Because HIF-1$\alpha$ is instable with oxygen, HIF-1 is scarce in normal mammalian cells. However, HIF-1$\alpha$ is expressed in pathological conditions such as cancer and obesity. Inhibiting HIF-1$\alpha$ may be of therapeutic value for these pathologies. Here, we investigated whether emodin, derived from the herb of Rheum palmatum L, which is also known as Chinese rhubarb, and is native to China, regulates HIF-1$\alpha$ expression. Male C57BL/6 mice without or with diet-induced obesity were treated with emodin for two weeks, while control mice were treated with vehicle. HIF-1$\alpha$ expression was determined by Western blot. We found that emodin inhibited obesity-induced HIF-1$\alpha$ expression in liver and skeletal muscle but did not regulate HIF-1$\alpha$ expression in the kidneys or in intra-abdominal fat. In vitro, emodin inhibited HIF-1$\alpha$ expression in human HepG2 hepatic cells and Y1 adrenocortical cells. Further, we investigated the mechanisms of HIF-1$\alpha$ expression in emodin-treated HepG2 cells. First, we found that HIF-1$\alpha$ had normal stability in the presence of emodin. Thus, emodin did not decrease HIF-1$\alpha$ by stimulating its degradation. Importantly, emodin decreased the activity of the signaling pathways that led to HIF-1$\alpha$ biosynthesis. Interestingly, emodin increased HIF-1$\alpha$ mRNA in HepG2 cells. This may be a result of feedback in response to the emodin-induced decrease in the protein of HIF-1$\alpha$. In conclusion, emodin decreases hepatic HIF-1$\alpha$ by inhibiting its biosynthesis.

Spinal cord injury is a devastating neurological disease in desperate need of a cure. We have previously shown that overexpression of the adhesion molecule L1 contributes to locomotor recovery after injury and were therefore interested in how electro-acupuncture would influence the expression of this molecule. Here, we investigated the effects of electro-acupuncture at “Jiaji” points (EX-B2), newly established by us, in young adult mice to determine whether improved recovery via electro-acupuncture could be due to enhanced L1 expression. Locomotor function, as evaluated by the Basso Mouse Scale score and by catwalk gait parameters, was improved by electro-acupuncture at different time points after injury in parallel with enhanced levels of L1 expression. Interestingly, the levels of the astrocytic marker glial fibrillary acidic protein (GFAP) were also increased, but only in the early phase after injury, being reduced at later stages during recovery. Acupuncture alone showed less pronounced changes in expression of these molecules. We propose that electro-acupuncture improves regeneration in part by promoting the L1 expression and beneficial activation of stem cells, and by differentially modulating the expression of GFAP by promoting regeneration-conductive astrocytic responses at initial stages and reducing regeneration-adversive activation in the secondary stages. Expression of the stem cell marker nestin was upregulated by electro-acupuncture in the acute stage. The combined observations show for the first time in mice the beneficial functions of electro-acupuncture at Jiaji points in the spinal cord injury mouse model and provide novel insights into some molecular mechanisms underlying electro-acupuncture in spinal cord injury.

Building on the work of Schimmerling [Coherent sequences and threads, Adv. Math.216(1) (2007) 89–117] and Steel [PFA implies AD^L(ℝ), J. Symbolic Logic70(4) (2005) 1255–1296], we show that the failure of square principle at a singular strong limit cardinal implies that there is a nontame mouse. The proof presented is the first inductive step beyond L(ℝ) of the core model induction that is aimed at getting a model of AD_ℝ + "Θ is regular" from the failure of square at a singular strong limit cardinal or PFA.

We prove the following result which is due to the third author. Let $n\ge 1$. If ${\mathrm{\Pi }}_n^1$ determinacy and ${\mathrm{\Pi }}_{n+1}^1$ determinacy both hold true and there is no ${\mathrm{\Sigma }}_{n+2}^1$-definable ${\omega }_1$-sequence of pairwise distinct reals, then $M_n^{\#}$ exists and is ${\omega }_1$-iterable. The proof yields that ${\mathrm{\Pi }}_{n+1}^1$ determinacy implies that $M_n^{\#}(x)$ exists and is ${\omega }_1$-iterable for all reals $x$. A consequence is the Determinacy Transfer Theorem for arbitrary $n\ge 1$, namely the statement that ${\mathrm{\Pi }}_{n+1}^1$ determinacy implies ${⅁}^{(n)}(<{\omega }^2-{\mathrm{\Pi }}_1^1)$ determinacy.

We establish natural criteria under which normally iterable premice are iterable for stacks of normal trees. Let $\mathrm{\Omega }$ be a regular uncountable cardinal. Let $m<\omega$ and $M$ be an $m$-sound premouse and $\mathrm{\Sigma }$ be an $(m,\mathrm{\Omega }+1)$-iteration strategy for $M$ (roughly, a normal $(\mathrm{\Omega }+1)$-strategy). We define a natural condensation property for iteration strategies, inflation condensation. We show that if $\mathrm{\Sigma }$ has inflation condensation then $M$ is ${(m,\mathrm{\Omega },\mathrm{\Omega }+1)}^{\ast }$-iterable (roughly, $M$ is iterable for length $\le \mathrm{\Omega }$ stacks of normal trees each of length $<\mathrm{\Omega }$), and moreover, we define a specific such strategy ${\mathrm{\Sigma }}^{\mathrm{\text{st}}}$ and a reduction of stacks via ${\mathrm{\Sigma }}^{\mathrm{\text{st}}}$ to normal trees via $\mathrm{\Sigma }$. If $\mathrm{\Sigma }$ has the Dodd-Jensen property and $\mathrm{\text{card}}(M)<\mathrm{\Omega }$ then $\mathrm{\Sigma }$ has inflation condensation. We also apply some of the techniques developed to prove that if $\mathrm{\Sigma }$ has strong hull condensation (introduced independently by John Steel), and $G$ is $V$-generic for an $\mathrm{\Omega }$-cc forcing, then $\mathrm{\Sigma }$ extends to an $(m,\mathrm{\Omega }+1)$-strategy ${\mathrm{\Sigma }}^{+}$ for $M$ with strong hull condensation, in the sense of $V[G]$. Moreover, this extension is unique. We deduce that if $G$ is $V$-generic for a ccc forcing then $V$ and $V[G]$ have the same $\omega$-sound, $(\omega ,\mathrm{\Omega }+1)$-iterable premice which project to $\omega$.

The data of frequency properties of single neurons in the auditory midbrain center, central nucleus of the inferior colliculus and morphologic aspects of its functional ordering are reviewed. On the basis of reconstruction of single units frequency receptive fields and morphophysiologic mapping of their location within the frequency-band lamina of the central nucleus the model of spectral coding by auditory midbrain neurons is developed. The main structural basement of spectral coding in the auditory midbrain is the tonotopic organization of its central nucleus as well as the order in its morphological structure expressed in alternation of layers of disc-shaped neurons and neuropil. The main neural mechanism of spectral coding is a critical bands mechanism. The critical bandwidth is controlled by inhibitory disc-shaped neurons which constitute an essential part of the population of the central nucleus disc-shaped cells.

Recently, a number of collaborative large-scale mouse mutagenesis programs have been launched. These programs aim for a better understanding of the roles of all individual coding genes and the biological systems in which these genes participate. In international efforts to share phenotypic data among facilities/institutes, it is desirable to integrate information obtained from different phenotypic platforms reliably. Since the definitions of specific phenotypes often depend on a tacit understanding of concepts that tends to vary among different facilities, it is necessary to define phenotypes based on the explicit evidence of assay results. We have developed a website termed PhenoSITE (Phenome Semantics Information with Terminology of Experiments: ), in which we are trying to integrate phenotype-related information using an experimental-evidence–based approach. The site's features include (1) a baseline database for our phenotyping platform; (2) an ontology associating international phenotypic definitions with experimental terminologies used in our phenotyping platform; (3) a database for standardized operation procedures of the phenotyping platform; and (4) a database for mouse mutants using data produced from the large-scale mutagenesis program at RIKEN GSC. We have developed two types of integrated viewers to enhance the accessibility to mutant resource information. One viewer depicts a matrix view of the ontology-based classification and chromosomal location of each gene; the other depicts ontology-mediated integration of experimental protocols, baseline data, and mutant information. These approaches rely entirely upon experiment-based evidence, ensuring the reliability of the integrated data from different phenotyping platforms.

The object of this paper is to present a set of techniques integrated into two low-cost human computer interfaces. Although the interfaces have many potential applications, one main application is to help the disabled persons to attain or regain some degree of independent communications and control. The first interface is a voice-controlled mouse and the second one is an accelerometer-based mouse.

The object of this paper is to present a low-lost vision-based computer interface which allows people with disabilities to use their head movements to manipulate computers. Our system requires only one low-cost web camera and a personal computer. Several experiments were conducted to test the performance of the proposed human-computer interface.

Genome-scale metabolic modeling is a systems-based approach that attempts to capture the metabolic complexity of the whole cell, for the purpose of gaining insight into metabolic function and regulation. This is achieved by organizing the metabolic components and their corresponding interactions into a single context. The reconstruction process is a challenging and laborious task, especially during the stage of manual curation. For the mouse genome-scale metabolic model, however, we were able to rapidly reconstruct a compartmentalized model from well-curated metabolic databases online. The prototype model was comprehensive. Apart from minor compound naming and compartmentalization issues, only nine additional reactions without gene associations were added during model curation before the model was able to simulate growth in silico. Further curation led to a metabolic model that consists of 1399 genes mapped to 1757 reactions, with a total of 2037 reactions compartmentalized into the cytoplasm and mitochondria, capable of reproducing metabolic functions inferred from literatures. The reconstruction is made more tractable by developing a formal system to update the model against online databases. Effectively, we can focus our curation efforts into establishing better model annotations and gene–protein–reaction associations within the core metabolism, while relying on genome and proteome databases to build new annotations for peripheral pathways, which may bear less relevance to our modeling interest.

In this chapter we discuss how to engineer 3D pulmonary tissue constructs in vitro using primary isolates of foetal mouse distal lung cells. When cultured in hydrogel-based 3D constructs, the mixed cell population, comprised epithelial, mesenchymal and endothelial cells, organised into alveolar forming unit (AFU)-like sacculated structures, which, in terms of morphology and cytodifferentiation, were reminiscent of native distal lung. By using a unique, serum-free medium supplemented with a cocktail of tissue-specific growth factors, we were able to induce concomitant alveolisation and neovascularisation when culturing the cells in the hydrogels, but not in scaffolds composed of synthetic polymers. Our data suggest that our in vitro model is capable of recapitulating the parallel morphogenesis of epithelial and endothelial pulmonary tissue components, which may occur through dynamic paracrine interactions. These results also stress the importance of the complex input from co-cultures, tissue-specific growth factors and integrin signalling for successful tissue engineering in vitro. In a mouse model in vivo, incorporation of the primary lung cell isolates into Matrigel plugs, implanted either subcutaneously (s.c.), or under the kidney capsule, leads to the formation of sacculated AFUs in close proximity to patent capillaries. Effective functional vascularisation, however, was only observed upon addition of angiogenic growth factors to the scaffolds and their controlled release over time. Use of a fluorescent cell tracker confirmed that the neovessels in the constructs comprised endothelial cells from both the host and the grafts. These data demonstrate that it is feasible to generate vascularised pulmonary tissue constructs in vivo with proper epithelial differentiation, and that the degree of vascularisation may be manipulated by incorporating the release of an angiogenic factor within the construct.

Intense sound exposure causes permanent hearing loss due to hair cell and cochlear damage. Prior conditioning with sublethal stressors, such as non-traumatic sound, heat stress and restraint protects ear from acoustic injury. To explore the mechanisms of conditioning-induced cochlear protection, Young's modulus and the amount of filamentous actin (F-actin) of mouse outer hair cells (OHCs) with heat stress and those without such stress were investigated by atomic force microscopy (AFM) and confocal laser scanning microscopy (CLSM), respectively. Heat stress caused an increase in Young's modulus of OHCs at 3–6 h after its application along with an increase in their amount of F-actin. These time courses are similar to the previous report in which heat stress was shown to suppress permanent threshold shift. These results indicate that heat stress structurally modifies OHCs to increase their F-actin and thereby renders them stiffer, resulting in protection of ear from acoustic injury.

Because the mammalian Y chromosome is largely isolated from the reciprocal recombination with a paired homolog that has underpinned the mapping and positional cloning of genes elsewhere in the genome, the identification of Y gene functions separable from that of testis determination has derived primarily from deletion mapping. Evidence for a Y gene function in spermatogonial proliferation mapping to the mouse Y chromosome short arm was first obtained in 1986, and functions in spermiogenesis to the long arm in 1988 and to a second deletion interval on the short arm in 1998. Defining the gene content of the deletion intervals has been a major task. The first short arm deletion interval proved to be >900 kb, and it was not until 1998 that the eight genes lying partially or completely within the interval were identified. The other two deletion intervals are much larger, and comprise regions of repetitive DNA within which there are multi-copy Y genes. Assigning the functions defined by the deletions to specific genes has proved extremely difficult because gene targeting is not an option for multi-copy genes, and has been unsuccessful for single-copy Y genes. Here, we review attempts to use transgene rescue to identify the Y gene(s) that underlies each of the deletion phenotypes. The only success has been in assigning the Y short arm gene function in spermatogonial proliferation to Eif2s3y. There is a closely related X-linked copy, Eif2s3x, which encodes a virtually identical protein (a subunit of the translation initiation factor EIF2); we have now shown that an Eif2s3x transgene also rescues the spermatogonial proliferation failure. Since Eif2s3x escapes X inactivation and is thus expressed from both X copies, the normal expressed Eif2 dosage is two in males and females; this suggests that the spermatogonial block in the deletion mice is due to haploinsufficiency for the EIF2 subunit. However, this still begs the question why spermatogonia are the only cells that are overtly compromised by this haploinsufficiency. Although the transgene rescue approach has so far only achieved this single success, analysis of the Eif2 transgene rescue mice has pointed to two further functions for genes mapping to the same deletion interval as Eif2s3y: one in relation to the spindle checkpoint at the first meiotic metaphase, and the second at the stage when sperm head elongation and tail development begin. Hopefully, it will not be long before the Y genes matching these two new functions are identified.