Age-related changes in bone mineral density, cross-sectional area and the strength of long bones in the hind limbs and first lumbar vertebra in female Wistar rats

Age-related changes in bone mineral density (BMD) and cross-sectional area and bone strength index (SSI) of the femur, tibia, humerus, and first lumbar vertebra in female Wistar (WM/MsNrs) rats were examined by a quantitative computed tomography (pQCT) method. One hundred and sixteen virgin female Wistar (WM/MsNrs) rats aged 2-33 months were used. The data indicate that the total BMD values of metaphyses and diaphyses of long bones increased until 12 months, then decreased to a varying degree depending on the bone after 15-24 months, but the values of cortical and trabecular BMD with age were not always similar to the total BMD value. Nevertheless, the values for cross-sectional area and SSI in the long bones increased regardless of the total BMD decrease with age, indicating that this increase might have been due to a characteristic of the modeling pattern in rats. The total and cortical BMD values in the first lumbar vertebra decreased after 18 months, and SSI did after 15 months. The data obtained in this study were compared with those obtained from males in a previous study. In conclusion, it was indicated that in this strain the rats over 12 months with the highest total BMD values in the femur and tibia, and before the onset of various tumors, are useful as a model animal for osteoporosis experiments and observation of senile bone change.     

Genetic diagnosis of claudin-16 deficiency and sex determination in bovine preimplantation embryos

Renal tubular dysplasia is an autosomal recessively inherited disorder in Japanese black cattle that is due to deletion mutations in the claudin-16 gene and causes chronic renal failure and death of affected animals. Here, we report a multiplex-PCR procedure to determine the genotype for claudin-16 deficiency in preimplantation embryos. The presence or absence of the wild-type and mutant allele(s) was precisely detected with the multiplex-PCR using as little as 5 pg of genomic DNA from leukocytes. When biopsied embryo cells were examined for claudin-16 deficiency, 97.2% of genotypes were consistent with the PCR results obtained for the corresponding embryos. In addition, sexing of embryos by PCR was performed using an aliquot of DNA extracted from biopsied embryo cells, and determination of claudin-16 genotype and sex was successfully achieved with an efficiency of 91.7% for claudin-16 genotyping and 83.3% for sexing. The production of a 100-day fetus that was male and homozygous for claudin-16 deficiency, as expected from the analysis of biopsied embryo cells, gave evidence of the reliability and applicability of this procedure for preventing the transmission of this disease and for enabling advances in animal breeding.     

Different effects on the inflammatory lesions in the lacrimal and salivary glands after neonatal thymectomy in IQI/Jic mice, a model for Sjögren's syndrome

Thymectomy on day 3 after birth (D3Tx) is understood to eliminate CD4+CD25+ regulatory T cells (Treg) from the peripheral T cell repertoire in rodents, leading to the activation of autoreactive T cells. Herein, D3Tx was performed in IQI/Jic mice, a model for Sj?gren's syndrome characterized by autoimmune infiltrations into the lacrimal and salivary glands. At the age of 16 weeks, very severe lesions were observed in lacrimal tissues from thymectomized mice, suggesting that Treg preserve their immunoregulatory function in young IQI/Jic mice. In contrast, salivary lesions were comparable in the D3Tx and control groups, raising the possibility that either salivary-specific Treg escaped elimination in thymectomized mice or spontaneous lesions in IQI/Jic mice developed independently of the tolerance through Treg.     

Malate dehydrogenase activities are lower in some types of peripheral leucocytes of dogs and cats with type 1 diabetes mellitus

The activities of the enzymes in the malate-aspartate shuttle were measured in peripheral leucocytes of spontaneous type 1 diabetic dogs and cats treated with insulin injections. In the diabetic dogs and cats, fasting plasma glucose concentrations were three- or fourfold greater than the control levels in spite of insulin injections and the activities of cytosolic malate dehydrogenase (MDH), one of pivotal enzymes in the malate-aspartate shuttle, were remarkably lower than the controls. Depressed expression of cytosolic MDH mRNA was confirmed by RT-PCR analysis in the diabetic animals. The cytosolic ratio of MDH/lactate dehydrogenase (LDH) activity (M / L ratio) in leucocytes of the diabetic animals was significantly lower than that of normal control animals. The smaller M / L ratio appeared to reflect depression of energy metabolism in the diabetic animals. Intrinsically lower and further decreased MDH activities may be factors that induce insulin resistance observed in diabetic cats.     

Hypoglycemia associated with disseminated metastatic tonsillar squamous cell carcinoma producing insulin‐like growth factor‐I in a Pomeranian dog

A 13‐year‐old spayed female Pomeranian dog was presented for persistent, severe hypoglycemia (37 mg/dL; reference interval [RI] 75‐128 mg/dL). Progressive nonregenerative anemia (hematocrit 23.3%‐15.9%; RI 37.0%‐55.0%) and severe thrombocytopenia (36 000/µL; RI 200‐500 000/µL) were also noted. The serum insulin concentration was low (0.24 ng/mL; RI 0.302‐1.277 ng/mL). Computed tomography revealed multiple splenic nodules (1‐6 mm in diameter) and several hepatic nodules (7.6, 12 mm in diameter). Ultrasound‐guided fine‐needle aspiration of the splenic and hepatic nodules revealed low numbers of epithelial cells with mild cellular atypia, suggestive of a metastatic epithelial tumor, but the primary site was unknown at that time. On careful oral examination under general anesthesia, an enlarged right tonsil was noted grossly, and histopathologic examination of the tonsil diagnosed squamous cell carcinoma. Bone marrow aspirates and biopsies of the splenic and hepatic nodules were performed; all samples were diagnosed as metastatic carcinoma on histopathologic examination. No nodules were present in the pancreas, despite careful palpation during exploratory laparotomy. On immunohistochemistry, the neoplastic cells were positive for cytokeratin AE1/3 and insulin‐like growth factor (IGF)‐I but were negative for chromogranin A, PGP9.5, insulin, and inconclusive for IGF‐II. This is the first report of a primary IGF‐I‐producing squamous cell carcinoma in the tonsil of a dog with metastases to bone marrow, liver, and spleen, resulting in hypoglycemia.     

Localization of CO2 source by a hexapod robot equipped with an anemoscope and a gas sensor

A hexapod robot was developed to explore an agricultural field and gather agronomical information such as soil nutrients and plant growth. In addition, a sensor device for the robot to track a gas source was developed. The sensor device consisted of a CO₂ gas sensor and an anemoscope. The gas sensor detected the concentration of CO₂ in the surroundings, and the anemoscope indicated the upwind direction from the robot. An indoor test was conducted during which the hexapod robot tracked windward direction using the sensor device. According to the results, the hexapod robot with the sensor device successfully localized a CO₂ source that was in the upwind direction. The robot paused on the spot under windless conditions, and walked windward as soon as the wind started blowing again.     

Hyaluromycin,a New Hyaluronidase Inhibitor of Polyketide Origin from Marine Streptomyces sp.

Hyaluromycin (1), a new member of the rubromycin family of antibiotics, was isolated from the culture extract of a marine-derived Streptomyces sp. as a HAase inhibitor on the basis of HAase activity screening. The structure of 1 was elucidated through the interpretation of NMR data for the compound and its 3″-O-methyl derivative in combination with an incorporation experiment with [1,2-¹³C₂]acetate. The compound’s absolute configuration was determined by the comparison of its circular dichroism (CD) spectrum with those of other rubromycins. Hyaluromycin (1) consists of a γ-rubromycin core structure possessing a 2-amino-3-hydroxycyclopent-2-enone (C₅N) unit as an amide substituent of the carboxyl function; both structural units have been reported only from actinomycetes. Hyaluromycin (1) displayed approximately 25-fold more potent hyaluronidase inhibitory activity against hyaluronidase than did glycyrrhizin, a known inhibitor of plant origin.     

Comparison of the growth traits of a commercial pioneer tree species,paper mulberry (Broussonetia papyrifera L. Vent.), with those of shade-tolerant tree species: investigation of the ecophysiological mechanisms underlying shade-intolerance

In rural areas of northern Laos, a commercially valuable pioneer tree species, paper mulberry (Broussonetia papyrifera L. Vent.) has been recommended for intercropping under plantations of commercial trees. However, less is understood about the growth of this pioneer tree species in the understorey and the mechanism underlying the shade intolerance. We measured growth characteristics for seedlings of paper mulberry under four light intensities. We compared the relative growth rates in aboveground biomass and standing leaf area (RGR_mass and RGR_leaf), light-capture efficiency, and seeding-level mass-based daily photosynthetic rates (A _mass) with those of field-grown seedlings of eight shade-tolerant species to identify factors potentially responsible for shade-intolerance. Most growth traits of the paper mulberry seedlings did not differ consistently from those of the shade tolerant species. The ecophysiological–architectural model software showed higher A _mass and RGR_mass capacity in paper mulberry than in shade-tolerant species. Despite their higher RGR_mass, paper mulberry seedlings had negative RGR_leaf under shaded conditions due to short leaf lifespan. The linear RGR_mass–RGR_leaf relationship for paper mulberry had a high RGR_mass intercept, indicating that a high RGR_mass was required to provide positive RGR_leaf. Progressive decreases in standing leaf area with time, and possibly photosynthesis, appear to be responsible for the shade-intolerance of paper mulberry. Although intercropping of paper mulberry has been suggested in the species’ native region, understorey cultivation of paper mulberry would only be possible with relatively open canopies.     

Dodeca-CLE peptides as suppressors of plant stem cell differentiation

In plants and animals, small peptide ligands that signal in cell-cell communication have been suggested to be a crucial component of development. A bioassay of single-cell transdifferentation demonstrates that a dodecapeptide with two hydroxyproline residues is the functional product of genes from the CLE family, which includes CLAVATA3 in Arabidopsis. The dodecapeptide suppresses xylem cell development at a concentration of 10(-11) M and promotes cell division. An application, corresponding to all 26 Arabidopsis CLE protein family members, of synthetic dodecapeptides reveals two counteracting signaling pathways involved in stem cell fate.