Sex identification of Japanese black bear,Ursus thibetanus japonicus,by PCR based on amelogenin gene

A method for sex identification of the Japanese black bear was examined using a polymerase chain reaction (PCR) and sequencing of a part of the amelogenin gene. This gene is located on the X and Y chromosomes, and there are 54 nucleotide deletions on the Y chromosome-specific gene. Forty-seven (26 male and 21 female) DNA samples and 23 (13 male and 10 female) DNA samples, respectively extracted from white blood cells and hairs of Japanese black bears were analyzed. The primers SE47 and SE48 from this X-Y homologous region were used in sex identification by PCR amplification. These primers amplified X- and Y-specific bands, which could be used to discriminate between sexes by a length polymorphism in all samples. We suggest that PCR amplification using the primers SE47 and SE48 is useful for sex determination of the Japanese black bear and could be applied to DNA analysis of small samples such as hairs.     

Effect of Gaseous Acetic Acid Treatment on Breadmaking Properties of Flour

Eleven different wheat flours of varying protein content were treated with gaseous acetic acid. Each wheat flour exhibited maximum breadmaking properties (bread height and specific volume) at different levels of gaseous acetic acid treatment. There was a relationship (r = +0.7384) between the level of gaseous acetic acid treatment required for maximum breadmaking properties and the protein content of the wheat flour. Wheat flours with higher protein contents were more resistant to the decrease in pH value due to acetic acid treatment.     

A cat with myelodysplastic syndrome by administration of the methylation inhibitor Azacytidine

A 5-year-old female cat with nonregenerative anemia and thrombocytopenia was diagnosed with myelodysplastic syndromes (MDS), since peripheral blood and bone marrow (BM) examination revealed various dysplasias and a blast ratio of 19%. Chemotherapy with azacytidine (AZA; 70–35 mg/m², 3–5 days, three cycles) and treatment with prednisolone, antibiotics, and vitamin K2, and blood transfusion were performed. On day 106, blast cells and dysplasia had decreased in the BM, and the cat remained alive for at least 1,474 days. This report is the first on feline MDS treated with AZA, suggesting appropriate drug dosage, interval and effective combination should be investigated and the pharmacological and cell biological mechanisms needs to be elucidated in the future.     

Zoogeographical barriers causing discontinuous osteometrical variations in the northern treeshrew skulls

Morphological variation of the skull was examined in the northern treeshrew (Tupaia belangeri) from various localities across Southeast Asia. Through a multivariate analysis, the treeshrews from South Vietnam exhibited distinct morphological characteristics compared to other populations from Thailand and Laos, and Malaysia. The plots of the specimens of North Vietnam are not randomly mixed with Thailand plots segregation in the scatteregrams of canonical discriminant analysis. Since the skulls of the population from North Vietnam were morphologically similar to those form central Laos and northern and northeastern Thailand, the zoogeographical barrier effect of Mekong River was not clearly confirmed. The population of the Kanchanaburi in western Thailand is clearly smaller in size compared to the other populations. The southern border of the distribution of this species is determined by the Isthmus of Kra or Kangar-Pattani Line. In the northern treeshrew, which is distributed from southern China to Bangladesh and southern Thailand, we have detected osteometrical geographical variation driven by geography. These results indicate that the skull morphology in the Tupaia glis-belangeri complex distinctively differs in South Vietnam, western Thailand, and southern Thailand. The zoogeographical barrier and factor separating these districts are expected to clarify in the future.     

Virucidal and Immunostimulating Activities of Monogalactosyl Diacylglyceride from Coccomyxa sp. KJ,a Green Microalga,against Murine Norovirus and Feline Calicivirus

Human noroviruses are the most common pathogens causing acute gastroenteritis and may lead to more severe illnesses among immunosuppressed people, including elderly and organ transplant recipients. To date, there are no safe and effective vaccines or antiviral agents for norovirus infections. In the present study, we aimed to demonstrate the antiviral activity of monogalactosyl diacylglyceride (MGDG) isolated from a microalga, Coccomyxa sp. KJ, against murine norovirus (MNV) and feline calicivirus (FCV), the surrogates for human norovirus. MGDG showed virucidal activities against these viruses in a dose- and time-dependent manner—MGDG at 100 μg/mL reduced the infectivity of MNV and FCV to approximately 10% after 60 min incubation. In the animal experiments of MNV infection, intraoral administration of MGDG (1 mg/day) exerted a therapeutic effect by suppressing viral shedding in the feces and produced high neutralizing antibody titers in sera and feces. When MGDG was orally administered to immunocompromised mice treated with 5-fluorouracil, the compound exhibited earlier stopping of viral shedding and higher neutralizing antibody titers of sera than those in the control mice administered with distilled water. Thus, MGDG may offer a new therapeutic and prophylactic alternative against norovirus infections.     

Effect of Gaseous Acetic Acid on Dough Rheological and Breadmaking Properties

Breads baked from wheat flours (protein contents 14.1–16.5% at 14.0% mb) that were pretreated with 2–3 mL of gaseous acetic acid per kg of wheat flour, showed maximum bread height and specific volume (cm³/g). Flour-water suspension and the crumb pH values were gradually decreased with increased amounts of acetic acid. Gas generation and dough expansion tests with bread dough showed that the addition of the same amount of acetic acid, which achieved maximum specific volume, also showed the highest rate of gas generation and dough expansion. However, increasing acetic acid decreased these values. Scanning electron microscope (Cryo-SEM) observation showed that the bread dough made from the same acetic acid-treated flour indicated continuum and no cracks in the dough matrix. Evaluation of mixograms showed the decrease of mixing stability with increased acetic acid levels. Viscosity and water binding capacity of flour-water suspensions were sharply increased by the addition of acetic acid at pH 5.0–3.5.     

Study of Three-Dimensional Structure of Wheat Starch Granules Stained with Remazolbrilliant Blue Dye and Extracted with Aqueous Sodium Dodecyl Sulfate and Mercaptoethanol Solution

Wheat starch granules were obtained from soft wheat flour by acetic acid fractionation (pH 3.5), and the starch was stained by reaction with Remazolbrilliant blue (RBB) dye. RBB-stained starch was extracted with 1% sodium dodecyl sulfate (SDS) and 1% 2-mercaptoethanol (ME) for 14.5 hr at room temperature. This extraction step was repeated five times (extracts 1–5). SDS-ME extracts were subjected to size-exclusion column chromatography, and comparisons of their profiles for specific absorbance at 650 nm (A₆₅₀) and carbohydrates were made. After high molecular weight (HMW) carbohydrates on the starch granule surface were extracted, HMW carbohydrates inside the granule appeared to be extracted. Finally, low molecular weight (LMW) carbohydrates near the granule surface were extracted. Phase-contrast light microscopy of the treated starch granules showed that all granules became transparent. Two different interior structures were observed. Scanning electron microscopy indicated that the granule was split into two parts at the equatorial groove. The interior of the granule showed two different areas: a central hole area and the surrounding stratified area. Extraction beyond five times with the same solvent dissolved the weak part of the granule structure and left two types of skeletal structures. The appearance of the skeletal structure of the granule surface was different from the appearance of interior structures.     

Abundance of Type II Diamonds

More than 1300 natural microdiamonds, mostly averaging 0.0015 carat, were studied for transparency to ultraviolet radiation, lambda2537. Of most of them, of this size, 16 percent were completely transparent (presumably type II); 27 percent, completely absorbent (presumably type I). The remainder transmitted partially. The number of type-II diamonds is unexpectedly very high; a considerable proportion of microdiamonds may begin life as type II, later incorporating absorbent type-I material.     

Induced Resistance in Hypocotyl of Cucumber by Infection with Colletotrichum lagenarium in Leaves

Localized infection of cucumber leaves with Colletotrichum lagenarium induced resistance against infection after challenge inoculation with pathogenic fungi, including C. lagenarium and Pythium ultimum var. ultimum. Systemic acquired resistance in the hypocotyl was observed when challenge inoculation was made 4 to 7 days after the first inoculation of cotyledons. Seven days after the first inoculation of a primary leaf, induced resistance against the challenge inoculation in the hypocotyl was also observed. However, the hypocotyl did not develop induced resistance when plants were challenged by a wound inoculation with P. ultimum. Received 9 June 1999/ Accepted in revised form 13 December 1999