Characteristic upregulation of glucose-regulated protein 78 in an early lesion negative for hitherto established cytochemical markers in rat hepatocarcinogenesis

Previously, we reported α(2)-macroglobulin (α(2)M) to be a novel marker characteristic of rat hepatocellular preneoplastic and neoplastic lesions negative for hitherto well-established markers. In the present study, we further examined other candidate markers with specificity for the same type of lesions. Glutathione S-transferase-placental form (GST-P)-negative hepatocellular altered foci (HAF) were generated using a two-stage (initiation and promotion) carcinogenesis protocol with N,N-diethylnitrosamine (DEN) and either Wy-14,643 or clofibrate, two peroxisome proliferators. Microarray analysis using total RNAs isolated from laser-microdissected GST-P-negative HAF (amphophilic cell foci) and adjacent normal tissues was conducted along with immunohistochemistry and real-time RT-PCR. Staining for glucose-regulated protein 78 (GRP78) was detected in GST-P-negative HAF and hepatocellular adenomas, and slightly increased GRP78 mRNA expression was observed in the lesions by real-time RT-PCR analysis. Thus, an early increase of GRP78 expression in hepatocarcinogenesis is likely a feature of the amphophilic subset of HAF.     

Distribution of benzimidazole-resistant strains of the onion gray-mold neck rot pathogens,Botrytis aclada and Botrytis allii,in Hokkaido,Japan

Botrytis aclada and Botrytis allii, associated with onion gray-mold neck rot and isolated in Hokkaido, were tested for sensitivity to benzimidazole. Of the B. aclada strains, 59% were highly resistant and the remaining 41% were sensitive; all strains of B. allii were sensitive. Resistant strains were widespread in Hokkaido. We analyzed the sequences of the β-tubulin gene of resistant strains and detected the replacement of glutamic acid (GAG) by lysine (AAG) at codon 198. This is the first report of benzimidazole resistance in B. aclada. This study revealed a difference in fungicide sensitivity between the two Botrytis species.

     

Changes in the blood lactate dehydrogenase measurements in canines and felines following the international standardization of the assay method

Lactate dehydrogenase (LDH) in blood is measured using the Japanese Society of Clinical Chemistry (JSCC) method in Japan and the International Federation of Clinical Chemistry (IFCC) method in other countries. In human clinical practice, the IFCC method replaced the JSCC method due to international standardization. Moreover, veterinary LDH measurement will also eventually shift to the IFCC method. However, the relationship between the IFCC and JSCC methods for LDH in various animals and whether they can be equated or not have not yet been investigated. This study aimed to present the changes in measurements in canines and felines after switching to the IFCC method. The plasma LDH activity of canines (N=177) and felines (N=115), who visited a secondary care veterinary clinic, was measured using the JSCC and IFCC methods. The IFCC/JSCC ratio was <1.0 in 85% of canines and 88% of felines, indicating that the IFCC method tended to show lower values than the JSCC method, presumably because LDH5 is dominant among the LDH isozymes in canines and felines. The increase in the systematic errors of both assays was in the high value range, with some samples exceeding the error tolerance from near the upper end of the reference range. When switching to the IFCC method for LDH measurement in canines and felines, each institution should consider whether the reference range and clinical diagnostic values established by the JSCC method are appropriate for continued use.     

Usability of normal force distribution measurements to evaluate asymmetrical loading of the back of the horse and different rider positions on a standing horse

Pressure measurement devices in equine sports have primarily focused on tack (saddle pads and saddle fitting methods). However, saddle pressure devices may also be useful in evaluating the interaction and distribution of normal forces between the horse and rider, including rider position and riding technique. This study examined the validity, reliability, repeatability and possibilities of using a saddle pressure device to evaluate rider position. All measurements were performed using a standing horse. Validity was tested by calculating the correlation coefficient between measured normal force and the weight of the rider. Repeatability was tested by calculating intra-class correlation coefficients. The use of normal force measurements to evaluate horse–rider interaction was tested by adding a known weight to saddle or rider and collecting measurements with the rider sitting in four different positions.The device was found to be valid and reliable for force measurements when the measurement device was not replaced. The system could be used to determine the expected differences with added weight and in different rider positions. The normal force distribution measurement device proved to be a valid and reliable tool for studying the interaction between a rider and a static horse provided it is positioned carefully and consistently relative to both the horse and the saddle.     

First report of whisker mold,a postharvest disease on citrus caused by <Emphasis Type="Italic">Penicillium ulaiense</Emphasis> (in Japan)

In March 2006, stored fruits of the medium-to-late-ripening citrus variety Shiranuhi ([Citrus unshiu × C. sinensis] × C. reticulata) were found to have a disease similar to blue mold. The fungus causing this disease differed distinctly from the well-known, blue mold agent, Penicillium italicum, because it formed whisker-like coremia measuring 1–8 mm. On the basis of the morphological characteristics and the phylogenetic analysis based on the nucleotide sequence of the β-tubulin gene, the fungus was identified as P. ulaiense. This is the first report of citrus whisker mold caused by P. ulaiense in Japan.     

Crown gall of tobacco caused by <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> biovar 1 in tobacco fields

Crown gall disease of tobacco was found in Iwate Prefecture, Japan in 1995. Ten bacterial isolates, obtained from the galls of tobacco, were identified as Agrobacterium tumefaciens (Smith and Townsend 1907) Conn 1942 biovar 1 based on their ability to induce galls on the 14 tested plants, including tobacco after needle-prick inoculation, and on 12 cultural, physiological, and biological characteristics. The growth of the causal organism was not inhibited in vitro by agrocin of A. radiobacter strain K84. This report is the first on the natural occurrence of crown gall caused by A. tumefaciens on tobacco plants.     

Genetic diversity of the entomopathogen<Emphasis Type="Italic">Verticillium lecanii</Emphasis> on the basis of vegetative compatibility

Forty-three isolates ofVerticillium lecanii from insects, phytopathogenic fungi and other substrates were tested for vegetative compatibility by observing heterokaryon formation among complementary nitrate-nonutilizing (nit) mutants.nit mutants were isolated from 42/43 strains examined. Twenty-one isolates were self-incompatible, and the remaining 21 isolates were divided into 14 vegetative compatibility groups (VCGs): ten containing only a single strain each, and the remaining four containing two to four isolates each. Members of isolates in each of these VCGs all shared the same IGS haplotype. Further, the isolates within a VCG were correlated with one another in part by fragment patterns of mt-LrDNA, -SrDNA, Bt-2 and H4 region, by PCR-RFLP and -SSCP, but not by dsRNA. Two isolates belonging to VL-J2 have high virulence to aphids, whereas strains from VL-J1 lack this character. These findings indicate that two VCGs (VL-J1 and -J2) may originate from two distinct clonal lineages. Alternatively, high VCG diversity and HSI frequency ofV. lecanii might be associated with an array of distinct lineages. These data not only suggest relationships among DNA polymorphisms, virulence, and VCG, but also demonstrate genetic heterogeneity ofV. lecanii. http://www.phytoparasitica.org posting Sept. 30, 2003.     

Cryopreservation of in vitro-grown meristems of strawberry (Fragaria × ananassa Duch.) by encapsulation-vitrification

Alginate-coated meristems from in vitro-grown strawberry (Fragaria × ananassa Duch.) were successfully cryopreserved following dehydration by a vitrification solution. Excised meristems from cold-hardened plantlets at 4 °C for two weeks in the dark were encapsulated into alginate-gel beads containing a mixture of 2 M glycerol plus 0.4 M sucrose. These encapsulated and cryoprotected meristems were dehydrated with a highly concentrated vitrification solution (designated PVS2) for 2 hr at 0 °C prior to a plunge into liquid nitrogen. Successfully encapsulated vitrified meristems remained green and then developed shoots within one week after plating without intermediary callus formation. The average rate of shoot formation of encapsulated vitrified meristems amounted to nearly 90%. The cryogenic protocol was successfully applied to four cultivars of strawberry. It was also confirmed that encapsulated vitrified meristems cooled to - 196 °C produced higher shoot formation than encapsulated dried meristems. Besides, the recovery growth was much earlier than the latter. The encapsulation-vitrification method is easy to handle and produces high levels of shoot formation. Thus, the protocol promises for cryopreservation of strawberry germplasm. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of the lateral growth rate on wood properties in fast-growing hardwood species

We investigated the feasibility of using several fast-growing tropical or subtropical hardwood species for timber production by measuring key wood qualities in relationship to the high rates of lateral growth. The trees tested were sampled from even-aged plantations of Acacia mangium, A. auriculiformis, hybrid Acacia (A. mangium × A. auriculiformis), Eucalyptus grandis, E. globulus, and Paraserianthes falcataria (Solomon and Java origin) that had already reached commercial harvesting age. The released strain of the surface growth stress (RS), xylem density (XD), microfibril angle (MFA), and fiber length (FL) were measured at the outermost part of the xylem at breast height in each tree. Results were then compared to the lateral growth rate (radius/age) at breast height, which provides a relative indicator of the amount of tree growth per year. Our findings indicated that RS was constant, regardless of lateral growth rate in each species. Similar results were observed for XD, MFA, and FL, with a few exceptions, suggesting that high growth rates do not intrinsically affect the wood properties of fast-growing tropical or subtropical species that have reached harvesting age. However, special attention must be paid to patterns of xylem maturation when developing plantations of such species.