Isolation of Mycobacterium spp. from milking buffaloes and cattle in Nepal

In Nepal, mycobacterial isolates obtained from the milk and feces of buffaloes and cattle that were positive for the single intradermal cervical tuberculin (SICT) tests were genetically identified. A total of 36 mycobacterial strains were isolated from 39% of the buffaloes (14 of 36) and 34% of the cattle (11 of 32). Of the 36 strains, 13 were identified as M. bovis, and these strains were isolated from 17% of the buffaloes (6 of 36) and 16% of the cattle (5 of 32). M. bovis was isolated from both the milk and feces of one buffalo and one cattle, the milk alone of three buffaloes and three cattle, and the feces alone of two buffaloes and one cattle. These results suggest that milking buffaloes and cattle infected with M. bovis exist in Nepal. The remaining 23 strains were atypical mycobacteria. A program for the elimination of bovine tuberculosis should be implemented as soon as possible, and the public health education and proper hygienic practices may be required.     

Curcuminoids and sesquiterpenoids in turmeric (Curcuma longa L.) suppress an increase in blood glucose level in type 2 diabetic KK-Ay mice

Turmeric, the rhizome of Curcuma longa L., has a wide range of effects on human health. The chemistry includes curcuminoids and sesquiterpenoids as components, which are known to have antioxidative, anticarcinogenic, and antiinflammatory activities. In this study, we investigated the effects of three turmeric extracts on blood glucose levels in type 2 diabetic KK-A(y) mice (6 weeks old, n = 5/group). These turmeric extracts were obtained by ethanol extraction (E-ext) to yield both curcuminoids and sesquiterpenoids, hexane extraction (H-ext) to yield sesquiterpenoids, and ethanol extraction from hexane-extraction residue (HE-ext) to yield curcuminoids. The control group was fed a basal diet, while the other groups were fed a diet containing 0.1 or 0.5 g of H-ext or HE-ext/100 g of diet or 0.2 or 1.0 g of E-ext/100 g of diet for 4 weeks. Although blood glucose levels in the control group significantly increased (P < 0.01) after 4 weeks, feeding of 0.2 or 1.0 g of E-ext, 0.5 g of H-ext, and 0.5 g of HE-ext/100 g of diet suppressed the significant increase in blood glucose levels. Furthermore, E-ext stimulated human adipocyte differentiation, and these turmeric extracts had human peroxisome proliferator-activated receptor-gamma (PPAR-gamma) ligand-binding activity in a GAL4-PPAR-gamma chimera assay. Also, curcumin, demethoxycurcumin, bisdemethoxycurcumin, and ar-turmerone had PPAR-gamma ligand-binding activity. These results indicate that both curcuminoids and sesquiterpenoids in turmeric exhibit hypoglycemic effects via PPAR-gamma activation as one of the mechanisms, and suggest that E-ext including curcuminoids and sesquiterpenoids has the additive or synergistic effects of both components.     

Determination of silver in soils by atomic absorption spectrometry

A method for the determination of Ag in soils using atomic absorption spectrometry is described. The method involves the extraction of Ag from soil by boiling with 6 M HC1 followed by separation of the extracted Ag into methylisobutylketone (MIBK) using sodium N, N-diethyldithiocarbamate (DDTC) as a complexing agent. Silver is determined in the MIBK by direct aspiration into a flame atomic absorption spectrophotometer. The detection limit (S/N=2) for this method is 0.0001 mg L^?1 for aqueous solution and 0.002 mg kg^?1 for soil. The Ag content of even unpolluted soils can be determined by this method. The determination of Ag using this method was shown to be unaffected by the presence of various ions in the soil. The method was able to recover nearly 100% of Ag added to soil and approximately the same amounts of soil Ag were determined using this method as with HF-H₂SO₄ decomposition. For 3 reference soils of the Canadian Certified Reference Materials Project (CCRMP), the Ag values obtained by this method were the same as the values determined by Ebarvia et al. (1988). The amounts of Ag in the soils sampled in the Ichi River basin and the Ichi River sediments were determined using this method. This area has been polluted by Cd, Cu, Pb, and Zn discharged from the Ikuno Mine and Smelter. The Ag values ranged from 0.27 to 6.89 mg kg^?1 which were much higher than the values of the unpolluted soils.     

Long-term postsurgical outcomes of mast cell tumors resected with a margin proportional to the tumor diameter in 23 dogs

We evaluated the postsurgical outcomes of cutaneous or subcutaneous mast cell tumors (MCTs, n=25) in 23 dogs, resected with lateral surgical margins proportional to the widest tumor diameter, including at least one facial plane. The tumor diameter range was 0.3–2.6 cm (median: 0.9 cm), and all were histologically diagnosed as Kiupel’s low-grade MCT. Resection was histologically complete in 20, close (deep margin) in three, and incomplete (deep margin) in two. No dogs developed local recurrence at the site of initial surgery during follow-up of 161–2,219 days (median: 976 days). These results suggest that resection of low-grade, relatively small MCTs with surgical margins proportional to the tumor diameter is a practical procedure with high success rates.     

Evaporative light-scattering analysis of sulforaphane in broccoli samples: Quality of broccoli products regarding sulforaphane contents

Broccoli sulforaphane has received attention as a possible anticarcinogen. Sulforaphane analysis is difficult due to the lack of a chromophore for spectrometric detection. Hence, we developed a method for determining sulforaphane by using high-performance liquid chromatography (HPLC) coupled with an evaporative light-scattering detector (ELSD). Sulforaphane was extracted from acid-hydrolyzed broccoli samples, followed by solid-phase extraction and reversed-phase HPLC. Sulforaphane was detected by ELSD and concurrently identified by electrospray ionization time-of-flight mass spectrometry. The recovery of sulforaphane from broccoli samples was above 95%. The detection limit was 0.5 mug. The present method was sensitive enough to determine sulforaphane in mature broccoli, broccoli sprouts, and commercial broccoli products. Sulforaphane concentration in broccoli sprout (1153 mg/100 g dry weight) was about 10 times higher than that of mature broccoli (44-171 mg/100 g dry weight). Therefore, the broccoli sprout is recommended as a source of sulforaphane-rich products. In contrast, we found that sulforaphane could not be detected in most of broccoli products, suggesting present commercial broccoli products having low quality.     

Symptoms and molecular characterization of apple dimple fruit viroid isolates from apples in Japan

Apple dimple fruit viroid was detected from an apple tree (‘Jonagold’) bearing apples with mild dapple apple symptom. The isolates in Japan were distinct from those in apples in Italy and China and in fig in Italy. Graft-inoculation experiments showed that the symptoms were variable depending on the cultivar, and the symptom on ‘Starking Delicious’ was virtually similar to those reported in Italy. Symptoms induced by apple dimple fruit viroid were similar in part to those by apple fruit crinkle viroid or apple scar skin viroid, indicating that they cannot be discriminated by symptoms on any specific variety.     

Trichomes: interaction sites of tomato leaves with biotrophic powdery mildew pathogens

The present study aimed to explore the possibility of using the type I trichomes of tomato (Solanum lycopersicum) to monitor the infection processes of powdery mildews by microscopy. Individual trichome cells of two tomato genotypes were inoculated with pathogenic and non-pathogenic powdery mildew species, Pseudoidium neolycopersici, Erysiphe trifoliorum and Podosphaera xanthii. On the trichome cells of the tomato cultivar Moneymaker, hyphae of the pathogenic Pseudoidium neolycopersici (isolates KTP-03 and KTP-04) grew vigorously; whereas hyphal growth of the non-pathogenic Erysiphe trifoliorum and Podosphaera xanthii ceased after appressorium formation, which was associated with papilla formation and hypersensitive cell death, respectively. Similar infection processes of the tested powdery mildews were seen in Moneymaker epidermal cells. Therefore, tomato trichomes are suitable for analysing, at individual cell level, the infection processes of different pathotypes of powdery mildews and for observing the cytological responses of plants by non-pathogenic powdery mildews. On the other hand, it was observed that both isolates KTP-03 and KTP-04 failed to produce conidiophores on the hyphae elongating on Moneymaker trichomes. Similarly, no conidiophores were produced on the hyphae elongating on trichomes of Solanum peruvianum LA2172, which is resistant to KTP-03 and susceptible to KTP-04. Interestingly, delayed cell death occurred in LA2172 epidermal cells, which were attacked by KTP-03 hyphae elongating from trichomes and conidiophores were formed on new hyphae growing from the leaf epidermal cells. Thus, leaf trichomes and epidermal cells of the wild tomato species LA2172 reacted differently to the avirulent isolate KTP-03.     

Progression of the dose-related effects of estrogenic endocrine disruptors, an important factor in declining fertility, differs between the hypothalamo-pituitary axis and reproductive organs of male mice

For the purpose of investigation of working mechanisms in endocrine disruptors, we evaluated the dose-related effects of fetal and/or neonatal exposure to an estrogenic compound on the male reproductive organs in adult mice, particularly with respect to gene expression of steroidogenic acute regulatory protein (StAR). The pregnant ICR mice were given subcutaneous injections of 10 micro g/day/animal of diethylstilbestrol (DES) to subject the fetal mice to in utero exposure (IUE). Subsequently, the newborn male mice were subjected to neonatal exposure (NE) by treatment with vehicle or 0.1-10 micro g/day/animal of DES. Fertility rates of each group were as follows: control, 100%; IUE only, 60%; IUE+NE 0.1 micro g, 25%; IUE+NE 1 micro g, 0%; IUE+NE 10 micro g, 0%. In general histology, germ cell layers in the seminiferous tubules were thinned in the group of IUE+NE 10 micro g. Hypoplasia of the Leydig cells, in which the staining intensity of eosin was diminished, was also observed in the groups of IUE+NE 0.1-10 micro g. The androgen receptor (AR) and estrogen receptor alpha (ERalpha) immunoexpression in the Leydig cells of IUE+NE 1-10 micro g was slightly lower than that in the controls. Long-term dysfunction of the hypothalamo-pituitary-testicular axis, including sustained hypoproduction of gonadotropin and testosterone, and altered expressions of steroid hormone receptors and StAR genes were observed. The hypothalamo-pituitary control of gonadotropin secretion may be affected by the smaller doses of estrogenic agents than the reproductive organs. Furthermore, the fertility rate in the male mice exposed to this estrogenic agent was closely correlated with the testosterone levels, and even more so with the rate-limiting factor of steroidogenesis, StAR. This finding suggests that endocrine disruptors have an important pronounced effect on StAR gene expression.