Photodynamic hyperthermal therapy with indocyanine green (ICG) induces apoptosis and cell cycle arrest in B16F10 murine melanoma cells

We examined the effects of photodynamic hyperthemal therapy (PHT), which is a combination of photodynamic therapy (PDT) and hyperthermia (HT), on the apoptosis and cell cycle progression of murine melanoma B16F10 cells. The percentage of apoptotic cell was determined by flow cytometry using fluorescein isothiocyanate (FITC)-conjugated Annexin V and propidium iodide (PI) double staining. The cell cycle analysis was performed by PI staining with flow cytometry. The expression of cyclins and heat shock protein 70 (Hsp70) were examined by a Western blotting analysis. PHT induces death in B16F10 cells, and PHT-mediated apoptosis occurred acutely and persistently in vitro. Our study demonstrated that PHT using indocyanine green (ICG) and near infrared (NIR) light source induces apoptosis and G0/G1 cell cycle arrest in the B16F10 cells.     

Watercore in Japanese pear. III. Changes in the activities of some enzymes relating to the degradation of cell walls and the accumulation of sugar

The relationship between cellular breakdown and the activities of some cell-wall degrading enzymes (endocellulase, exocellulase, polygalacturonase, pectin methylesterase and β-galactosidase) was investigated in the watercored tissue of Japanese pear fruit, Pyrus serotina Rehder var. ‘Culta’ Rehder, cultivar ‘93-3’. In watercored tissue at an early stage, endocellulase activity, especially neutral form, was higher than that in healthy tissue. As the symptom of the watercore developed, polygalacturonase and β-galactosidase activities in the watercored tissue increased compared with those in healthy tissue. This indicated that in tissues at an early stage of watercore, these cell-wall degrading enzymes were activated, and operative in the breakdown of the cell wall. However, exocellulase and pectin methylesterase were not activated in the watercored tissue. The breakdown of cellular tissue was also supported by the fact that the DNA and protein contents decreased in the watercored tissue. Invertase activity decreased in the watercored tissue, compared with healthy tissue, and it was suggested that its decrease could be one of the causes of sucrose accumulation in the watercored tissue.     

Responses of leaf photosynthesis and plant growth to altered source–sink balance in a Japanese and a Dutch tomato cultivar

We investigated the effects of altered source/sink ratio by leaf or fruit pruning on leaf photosynthetic characteristics and whole-plant growth of ‘Momotaro York’, a Japanese cultivar, and ‘Dundee’, a Dutch cultivar and verified a hypothesis for sink-limitation of plant growth proposed by Tanaka and Fujita (1974). Plants were grown hydroponically with a high-wire system in a greenhouse for 11 weeks. Light-saturated photosynthesis of young, fully expanded leaves measured at atmospheric CO₂ partial pressure of 37 Pa and at an intercellular CO₂ partial pressure of 20 Pa was not influenced by alteration of source/sink ratio for either cultivar. Although soluble sugars were accumulated in leaves under high source/sink conditions, the amount of ribulose-1,5-bisphosphate carboxylase/oxygenase, a rate-limiting factor for CO₂-limited photosynthesis, was not affected by the altered source–sink balance. Net assimilation rate of a whole plant increased with decreasing source/sink ratio, but this can be accounted for by the changes in light interception per unit leaf area, without taking the source–sink relationship into consideration. It was concluded that the altered source/sink ratio did not change leaf photosynthetic capacity and the sink-limitation hypothesis cannot be applied to either cultivar under the conditions of the present study.     

Growth characteristic and sink strength of fruit at different CO2 concentrations in a Japanese and a Dutch tomato cultivar

The aim of the present study was to elucidate how fruit growth was limited by the source and sink capacities in a Japanese (‘Momotaro York’) and a Dutch (‘Dundee’) tomato cultivar. The two cultivars were grown hydroponically with a high-wire system in greenhouses for 25 weeks, and the growth characteristics and sink strength of fruit were determined. Fruits were pruned to four (4F) or one (1F) per truss. The latter were used to determine potential fruit growth, an indicator of fruit sink strength. Growth was also determined under normal (LC) and enriched (HC, 700 μmol mol⁻¹) CO₂ concentrations to examine the effect of source enhancement on fruit production. In both cultivars under normal CO₂, the growth rate of fruit pruned to 4F per truss was lower than that in 1F, indicating that maximum potential fruit growth was not achieved. Under HC conditions, fruit growth rate of ‘Dundee’ achieved in 4F trusses was lower than that in 1F. In ‘Momotaro York’ in HC, fruit growth in 4F trusses was close to potential. This implies that fruit growth was source-limited irrespective of CO₂ concentrations in ‘Dundee’ cultivar while fruit growth in ‘Momotaro York’ under normal and enriched CO₂ conditions was limited by source and sink strengths, respectively. Adjustments of cultural practices including increasing fruit number per truss and/or genetic approaches to enhancing fruit sink strength by breeding may improve fruit yields of Japanese cultivars under high source/sink conditions.     

Synovial fluid total protein concentration as a possible marker for canine idiopathic polyarthritis

Idiopathic polyarthritis (IPA) is a very common inflammatory arthropathy in the dog. Canine IPA is diagnosed mainly by detecting increased number of leukocytes in the synovial fluid (SF), which is easily influenced by glucocorticoid therapy. We obtained 31 SF samples from 24 IPA dogs prior to (n=19) and/or after (n=12) 1 to 10 weeks of glucocorticoid therapy. The SF total protein concentrations of IPA dogs were significantly higher than those of dogs with non-arthritis diseases (n=34) and healthy controls (n=10). Our data revealed that the SF total protein concentrations are not influenced by several weeks of glucocorticoid therapy. Hence, the SF total protein concentration is applicable as a diagnostic marker of canine IPA even when the patients are receiving glucocorticoid therapy.     

Comparison of fertility on intrauterine insemination between cryopreserved ejaculated and cauda epididymal sperm in dogs

The fertility was compared between ejaculated and cauda epididymal sperm sensitized with prostatic fluid in dog after freeze-thawing using the fertility of ova from the contralateral ovary after injection (2 × 10(8) sperm) into dog uterus on the unilateral ovariectomized side, on the basis of the presence or absence of conception. No significant difference was observed in sperm quality after freeze-thawing between the two groups and conception rates were equivalent and low. Therefore, to achieve a high fertility by intrauterine insemination of canine frozen-thawed ejaculated and cauda epididymal sperm, intrauterine insemination on both sides is recommended, rather than insemination with a lot of sperm of the uterine horn on one side.     

Differential spread of discoloured and non‐conductive sapwood among four Fagaceae species inoculated with Raffaelea quercivora

To reveal the relationship between the susceptibility of Fagaceae species to Raffaelea quercivora Kubono et Shin‐Ito and the tangential expansion of regions of discoloured and non‐conductive sapwood among the species, we inoculated branches of three Quercus species and one Castanopsis species with the fungus. The sapwood around the inoculation hole in all four species became non‐conductive in response to the infection before the discolouration. The expansion of the region of non‐conductive sapwood ceased within 2 weeks after the inoculation. The region of non‐conductive sapwood in Q. crispula and Q. serrata was larger than that in Q. glauca and C. cuspidata var. sieboldii. These results suggest that the region of non‐conductive sapwood expanded soon after the infection by R. quercivora. It was also clear that the transverse non‐conductive sapwood has close relationship with the susceptibility among Fagaceae species to R. quercivora.     

Flowering Date1, a major photoperiod sensitivity gene in adzuki bean,is a soybean floral repressor E1 ortholog

Adzuki bean is an important legume crop originating in temperate regions, with photoperiod in sensitivity being a key factor in its latitudinal adaptation. The Flowering Date1 (FD1) gene has a large effect on the photoperiodic response of flowering time, but the molecular basis for the effect of this locus is undetermined. The present study delimited the FD1 locus to a 17.1 kb sequence, containing a single gene, an E1 ortholog (VaE1). A comparison between Vigna angularis ‘Shumari’ (photoperiod insensitive) and ‘Acc2265’ (photoperiod sensitive) identified 29 insertions/deletions and 178 SNPs upstream of VaE1 in the FD1 locus. VaE1 expression in ‘Acc2265’ was greater under long-day than short-day conditions, whereas VaE1 expression in ‘Shumari’ was lower regardless of day length. These findings suggested that responsible gene of FD1 is a VaE1, which acts as a floral repressor by being upregulated in response to long-day conditions. The inability to upregulate VaE1 under long-day conditions was linked to its ability to flower under these conditions. These results provide greater understanding of the molecular control of a flowering date and clues enabling the breeding of adzuki bean at higher latitudes.