Multi-locus sequence analysis reveals host specific association between Bartonella washoensis and squirrels

To clarify phylogenetic relationships and genetic diversity among Bartonella washoensis strains obtained from squirrels, multi-locus sequence analysis (MLSA) with the 16S rRNA, ftsZ, gltA, groEL, ribC, and rpoB genes was applied for 20 strains of B. washoensis isolated from five genera of squirrels (Tamias, Tamiasciurus, Glaucomys, Sciurus, and Spermophilus) within the family Sciuridae. Sequence similarities in the concatenated sequences of B. washoensis strains from squirrels of different genera ranged from 94.7% (Sciurus vs. Spermophilus) to 98.4% (Tamiasciurus vs. Glaucomys). Phylogenetic trees based on the concatenated sequences revealed that B. washoensis strains formed five distinct clades and each clade correlated with the genus of squirrel from which the strains were originally obtained. The discrimination was supported by 100% bootstrap values and posterior probabilities, respectively. These results suggest that B. washoensis strains may have co-speciated with their squirrel hosts and provide new insights into the application of the MLSA to identify sources of B. washoensis infection with accuracy.     

Effect of Platelet-Rich Plasma-Incorporated Gelatin Hydrogel Microspheres and Subchondral Drilling on Equine Cartilage Defects

We evaluated the effect of platelet-rich plasma-incorporated gelatin hydrogel microsphere (PRP/GM) together with subchondral drilling for cartilage regeneration in horses. Cartilage defects (diameter, 4.5 mm; depth, 3.0 mm) were created in both third carpal bones of six Thoroughbred horses. A subchondral hole was drilled at the center of this cartilage defect (diameter, 2.0 mm; depth, 35 mm) in one randomly selected carpal joint (test joint), whereas no subchondral hole was made in the contralateral joint (control joint). Thereafter, the test joint was injected with PRP/GM and the control joint with saline-infused GM. Additional injections of PRP/GM were given 14 and 28 days after surgery. After 16 weeks, histologic analysis was conducted. The macroscopic and histologic scores of the test group were significantly higher than those of the control group (P < .05). These data indicated that the application of PRP/GM together with subchondral drilling accelerated cartilage regeneration in horses.     

Cyanobacteria from Terrestrial and Marine Sources Contain Apoptogens Able to Overcome Chemoresistance in Acute Myeloid Leukemia Cells

In this study, we investigated forty cyanobacterial isolates from biofilms, gastropods, brackish water and symbiotic lichen habitats. Their aqueous and organic extracts were used to screen for apoptosis-inducing activity against acute myeloid leukemia cells. A total of 28 extracts showed cytotoxicity against rat acute myeloid leukemia (IPC-81) cells. The design of the screen made it possible to eliminate known toxins, such as microcystins and nodularin, or known metabolites with anti-leukemic activity, such as adenosine and its analogs. A cytotoxicity test on human embryonic kidney (HEK293T) fibroblasts indicated that 21 of the 28 extracts containing anti-acute myeloid leukemia (AML) activity showed selectivity in favor of leukemia cells. Extracts L26-O and L30-O were able to partly overcome the chemotherapy resistance induced by the oncogenic protein Bcl-2, whereas extract L1-O overcame protection from the deletion of the tumor suppressor protein p53. In conclusion, cyanobacteria are a prolific resource for anti-leukemia compounds that have potential for pharmaceutical applications. Based on the variety of cellular responses, we also conclude that the different anti-leukemic compounds in the cyanobacterial extracts target different elements of the death machinery of mammalian cells.     

Age structure of wind-felled canopy trees for Siebold’s beech (<Emphasis Type="Italic">Fagus crenata</Emphasis>) in the northernmost population in Karibayama,Hokkaido

To determine the lifespan of Siebold’s beech (Fagus crenata), a dominant canopy tree species of the cool temperate forests of Japan, we estimated the age of 33 mature beech trees felled by a large typhoon in September 2004 at Karibayama, in southern Hokkaido. The study site lies within one of the northernmost distributions of Siebold’s beech populations in Japan. We counted tree rings from tree disks at a height of 1 m, and diameter growth was measured in annual increments. The sample age revealed two major peaks at 240 and 175 years. The diameter growth was good between 40 and 60 years of age and gradually decreased after 70 years of age. Annual diameter increments were noticeably small in good fruiting years, indicating a large allocation of annual carbon gains to the reproductive organs. Bud scale counts of juvenile trees revealed that the age at which the height reached 1 m was approximately 6–13 years. The lifespan of Siebold’s beech in the Karibayama population was therefore estimated to be approximately 250 years, an estimation that is higher than that previously reported for this species in the northernmost populations.     

Linkage analysis among loci for RAPDs, isozymes and some agronomic traits in Brassica campestris L.

Common bean, Phaseolus vulgaris L. is intercropped or relay cropped with maize in many Andean highlands of Colombia and Peru. Breeding beans for the target multiple cropping systems is essential for the development of productive and sustainable agriculture for the Andean smallholders. Outline of the breeding programme should follow the farming system approach with the establishment of on-farm trials and early farmers involvement. Bean breeding is oriented to minimize intercrop competition and to stabilize complementarity with maize. Genetic traits needed for improved varieties are divided as follows : traits not interacting with the cropping systems, traits specific to intercrops and traits related with socioeconomic and seed quality aspects. Screening, prebreeding and recombination nursery are better made under sole cropping while varietal improvement and on-farm trials are conducted under the target multiple cropping systems. Breeding schemes may involve recurrent, pedigree and bulk hybrid selection. The given application concerns the genetic improvement of P. coccineus, P. polyanthus and interspecific hybrids of P. vulgaris for both simultaneous and relay intercropping in Colombia and Peru. Earliness, cold tolerance, resistance to fungus diseases (mainly Ascochyta leaf blight and anthracnosis) and seed yield potential were the major objectives of the bean improvement programme. Priority has been given to the exploitation of the large diversity available in the secondary gene pool of common bean. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic nature of low capsaicin content in the variant strains induced by grafting in Capsicum annuum L.

Summary The genetic nature of low capsaicin content in variant strains induced by grafting was elucidated by crossing the G₅S₂₃ strain with the two cultivars, Yatsubusa (Capsicum annuum L. var. fasciculatum Ilish) and Spanish Paprika (C. annuum L. grossum Sendt) originally used for grafting. Decreased capsaicin synthesis was a stable characteristic for at least several generations from G₅S₁₆ to G₅S₂₃ and was transmitted to the progenies of crosses. This fact shows that the decreased capsaicin synthesis is a genetic trait in the graft-induced variant strains. The genetically dependent characteristic of pungency of the G₅S₂₃, Yatsubusa and Spanish Paprika were analysed by High Perfomance Liquid Chromatography measurement and tasting assay.Cross experiment between the G₅S₂₃ strain, hot parent Yatsubusa and sweet parent Spanish Paprika showed that pungent and sweet for pungency chracteristics are generally controlled by at least two pairs of genes. Although segregation of capsaicin content and pungency was clearly demonstrated in F₂ progenies from reciprocal crosses between the G₅S₂₃ strain and Spanish Paprika, F₂ progenies from reciprocal crosses between G₅S₂₃ and Yatsubusa were all hot and did not develop low capsaicin plants.     

Detection of giant myofibrillar proteins connectin and nebulin in fish meat by electrophoresis in 3-5 gradient sodium dodecyl sulfate polyacrylamide slab gels

An improved method was investigated for sodium dodecyl sulfate polyacrylamide slab gel electrophoresis (SDS-PAGE) to facilitate the analysis of the giant myofibrillar proteins, connectin and nebulin, in fish meat by using jack mackerel (Trachurus japonicus) as the sample fish. It was established that separation of the alpha-connectin band from the beta-connectin band by SDS-PAGE could be achieved by using 3-5% gradient gels with glycerol to facilitate the formation of a gradient with polymerization at 35 degrees C. SDS-PAGE samples of white dorsal muscle from the jack mackerel were homogenized with a 2% SDS solution containing an inhibitor mixture (1 microg/mL of phenylmethanesulfonyl fluoride, 1 microg/mL of leupeptin, and 1 microg/mL of E-64) and heated at 50 degrees C for 20 min. Heating these samples at 100 degrees C for 2 min resulted in the disintegration of connectin but did not affect nebulin. A purified myofibril sample and a whole muscle sample showed similar changes in the overall rate of degradation of whole connectin and nebulin during the postmortem storage period, but it was clear that beta-connectin was cleaved from alpha-connectin during the preparation of myofibrils at the early stage postmortem. Storage of the SDS-PAGE samples at -85 degrees C was preferable to storage at -18 degrees C for a long period.     

Predicting the saturated hydraulic conductivity of compacted subsoils using the non-similar media concept

It is widely recognized that saturated hydraulic conductivity is dominated by the micromorphology of soil pores rather than by the merely total porosity or dry bulk density. Nevertheless, some researchers are reporting that the decrease in saturated hydraulic conductivity of subsoil is simply associated with the decrease in soil porosity or increase in dry bulk density. Based on these understandings in published papers and on our preliminary field investigation, we assumed that micromorphology of soil pores in topsoils is subjected to be destroyed with continuous disturbance by frequent tillage while subsoils tend to be compacted without serious changes of micromorphology of soil pores. Thus, we focused on finding the dependence of saturated hydraulic conductivity on dry bulk density by separating the soils into tilled layer and compacted layer. The objective of this study was to describe the relationship between saturated hydraulic conductivity and dry bulk density using a theoretical model, the non-similar media concept (NSMC) model, capable of predicting saturated hydraulic conductivities of soils with different values of dry bulk densities. The study area was located near the Tone River in Saitama Prefecture, Japan, where the soils were classified into Haplic Brown Lowland Soils according to the Classification of Cultivated Soils in Japan (Eutric Fluvisol according to FAO/UNESCO). Two sites, where the topsoils were seasonally tilled while the subsoils were sustained as it is, and another site where the topsoil was seasonally tilled, too, but extra deep tillage (1 m tillage depth) had been done, were chosen for the measurements. The saturated hydraulic conductivities and dry bulk densities of undisturbed soil cores from different depths were measured in the laboratory. The NSMC model was carefully applied only when the soil textures were the same among samples. The well-known conventional equations formulated by Kozeny–Carman and by Campbell, were used to compare the applicabilities with the NSMC model. The NSMC model succeeded in predicting the saturated hydraulic conductivities in the compacted subsoils. On the other hand, the NSMC model was not applicable to the tilled topsoils and to the deeply tilled subsoil. The saturated hydraulic conductivity of tilled topsoils and deeply tilled subsoil was always lower than that of compacted subsoils at the same dry bulk densities. The Kozeny–Carman and Campbell equations both failed in the prediction of saturated hydraulic conductivity in subsoil. It was concluded that the saturated hydraulic conductivity of subsoils under compaction without extreme disturbance is well related with its dry bulk density by the NSMC model.