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.     

Genetic diversity of Pinus densiflora pollen flowing over fragmented populations during a mating season

We attempted to evaluate the genetic diversity of long-distance transported pollen flowing over fragmented Pinus densiflora populations during a mating season. A P.?densiflora clonal seed orchard, which was located in a rural area where many fragmented populations exist, was selected for pollen capture. Immigrant pollen captured by three clones having different flowering times was regarded as the pollen flowing over fragmented populations during a mating season. The genetic diversity (H _e) values of the immigrant pollen captured by the three clones were high (H _e?>?0.894). The correlation of paternity (r _p) values of the seeds having immigrant parent generated from the three clones were calculated to be negative. From these parameters, the pollen cloud is considered to have maintained high genetic diversity during the mating season. The genetic composition of the pollen cloud showed slight variation. The pollen captured by different trees (i.e., clonal ramets of the three clones) was significantly different based on analysis of molecular variance. Especially, the pollen pools captured by trees planted in the western side of the orchard were significantly different from the gene pool of the surrounding populations. Factors affecting this differentiation could be that the donors of the pollen transported to the orchard vary with time, as well as nonuniform dispersal of the pollen. From these results, the pollen flowing over fragmented P.?densiflora populations is considered to have high genetic diversity, compensating to some extent for fragmentation.     

Heterotrophic respiration causes seasonal hysteresis in soil respiration in a warm-temperate forest

To assess the effect of changes in organic litter stock on seasonal changes in heterotrophic respiration (R _H), soil respiration (R _S), and total ecosystem respiration (R _E), we measured seasonal changes in leaf litter respiration (R _LL) by the chamber method and estimated the seasonal change in total R _H using the RothC model in a warm-temperate mixed deciduous?Cevergreen forest in Japan. Both R _E and R _S had seasonal hysteresis and were higher in spring than at the same temperature during autumn. Under warm and humid conditions, the rate of decomposition of newly supplied leaf litter in one?year was high (60% loss). Consequently, R _LL and R _H were higher in spring after leaf drop, when more fresh material was available, than in autumn. In this study, 42 and 88% of the difference in R _E and R _S between spring and autumn (soil temperature 16?C18°C) could be accounted for by the difference in R _H, respectively, and 71% of the difference in R _H could be accounted for by the difference in R _LL. This study showed that seasonal changes in heterotrophic respiration (R _LL and R _H) could be a major factor in the seasonal hysteresis of R _E and R _S.     

Use of image analysis to estimate anthocyanin and UV-excited fluorescent phenolic compound levels in strawberry fruit

Strawberry is rich in anthocyanins, which are responsible for the red color, and contains several colorless phenolic compounds. Among the colorless phenolic compounds, some, such as hydroxycinammic acid derivatives, emit blue-green fluorescence when excited with ultraviolet (UV) light. Here, we investigated the effectiveness of image analyses for estimating the levels of anthocyanins and UV-excited fluorescent phenolic compounds in fruit. The fruit skin and cut surface of 12 cultivars were photographed under visible and UV light conditions; colors were evaluated based on the color components of images. The levels of anthocyanins and UV-excited fluorescent compounds in each fruit were also evaluated by spectrophotometric and high performance liquid chromatography (HPLC) analyses, respectively and relationships between these levels and the image data were investigated. Red depth of the fruits differed greatly among the cultivars and anthocyanin content was well estimated based on the color values of the cut surface images. Strong UV-excited fluorescence was observed on the cut surfaces of several cultivars, and the grayscale values of the UV-excited fluorescence images were markedly correlated with the levels of those fluorescent compounds as evaluated by HPLC analysis. These results indicate that image analyses can select promising genotypes rich in anthocyanins and fluorescent phenolic compounds.     

Improvement of DNA extraction method for dried blood spots and comparison of four PCR methods for detection of Babesia gibsoni (Asian genotype) infection in canine blood samples

To eradicate canine babesiosis in epidemic areas, mass-screening of the infection situation of Babesia gibsoni including occult infection is necessary. The development of cost-effective method for storage and transport of blood samples is required. A highly efficient DNA extraction procedure from dried blood spots (DBS) onto Whatman 3MM filter paper was developed for the diagnosis of B. gibsoni infection in dog by PCR. In 3 extraction methods, Chelex-based method in combination with saponin washing and phenol-chloroform-isoamyl alcohol extraction (Saponin-PCI method) provided the best results. Sensitivity of the 4 previously described PCR methods for detection of B. gibsoni infection was also compared using serially diluted blood samples of B. gibsoni-infected dogs. The PCR method using Gib599F/Gib1270R primer pair provided the best performance. To evaluate the stability of DNA in DBS, DBS of B. gibsoni-infected dogs stored at room temperature for 2 months. The stability was superior to whole blood samples stored at -20 degrees C for 2 months. This highly efficient DNA extraction method on DBS using Whatman 3MM filter paper has potential to be cost-effective and high performance tool for storage, and molecular diagnosis of clinical blood sample from dog. This procedure in combination with the PCR method using Gib599F/Gib1270R primer pair may greatly assist in diagnosis of B. gibsoni infection in dog populations that are geographically distant.     

Anther culture in rice proportionally rescues microspores according to gametophytic gene effect and enhances genetic study of hybrid sterility

Background

To investigate plant hybrid sterility, we studied interspecific hybrids of two cultivated rice species, Asian rice (Oryza sativa) and African rice (O. glaberrima). Male gametes of these hybrids display complete sterility owing to a dozen of hybrid sterility loci, termed HS loci, but this complicated genetic system remains poorly understood.

Results

Microspores from these interspecific hybrids form sterile pollen but are viable at the immature stage. Application of the anther culture (AC) method caused these immature microspores to induce callus. The segregation distortion of 11 among 13 known HS loci was assessed in the callus population. Using many individual calli, fine mapping of the HS loci was attempted based on heterozygotes produced from chromosome segment substitution lines (CSSLs). Transmission ratio distortion (TRD) from microspores was detected at 6 of 11 HS loci in the callus population. The fine mapping of S₁ and S₁₉ loci using CSSLs revealed precise distances of markers from the positions of HS loci exhibiting excessive TRD.

Conclusions

We demonstrated that AC to generate callus populations derived from immature microspores is a useful methodology for genetic study. The callus population facilitated detection of TRD at multiple HS loci and dramatically shortened the process for mapping hybrid sterility genes.

     

Quantifying landscape pattern and ecosystem service value changes in four rapidly urbanizing hill stations of Southeast Asia

Context

Hill stations are known for their favorable cool climate and natural environments which generate valuable ecosystem services that benefit the local population, tourists and visitors. However, rapid urbanization threatens the sustainability of these highly valued fragile landscapes.

Objectives

We aim to characterize and quantify the changes in the landscape patterns and ecosystem service values (ESVs) of Baguio (Philippines), Bogor (Indonesia), Dalat (Vietnam), and Pyin Oo Lwin (Myanmar), and discuss their implications to landscape sustainability.

Methods

We used remote sensing imagery to map land-use/cover (2001 and 2014), and spatial metrics and gradient analysis to characterize the changes in landscape pattern. We employed a benefit transfer method to estimate the changes in ESV and human-to-ESV ratio. A land-change model was used to simulate different scenarios of future built-up expansions (2014–2030).

Results

The landscapes of Dalat and Pyin Oo Lwin are becoming more fragmented, while those of Baguio and Bogor are getting more aggregated. Dalat had the highest decrease (absolute change) in ESV and H-ESV ratio, while Bogor had the highest percentage decrease (2001–2014).

Conclusions

Rapid urbanization has been a major factor in the landscape transformation of Baguio, Bogor, Dalat and Pyin Oo Lwin. If the current built-up expansion rate will speed up, the decline in future ESV and H-ESV ratio (2014–2030) will be higher than if the rate will continue or slow down. Unless the concept of landscape sustainability is taken seriously in landscape and urban planning, the respective ‘values’ of these precious hill stations will become less and less.

     

Detection of genetic variation among seed and seedlings ofChamaecyparis obtusa using allozyme markers

The effects of nursery practices on genetic composition and structure were studied inChamaecyparis obtusa by analyzing seeds and one-, two-, and three-year-old seedlings. Enzyme polymorphisms of the6Pgd-2, G6pd, Got, Gk, Shd-2, Pod, Dia-1, andPgm loci were used. OnlyDia-1 indicated significantly different allelic frequencies between seeds and three-year-old seedlings. Most of the genetic diversity of the parental clones in this seed orchard could be transferred to the offsprings. Genetic diversity on the basis of an average expected heterozygosity did not differ between seeds and seedlings. A slight excess of homozygotes in seeds and a significant excess of heterozygotes in seedlings of different ages to the expected panmictic proportions were found. Most of the differences in genotypic distributions among the seed and seedling stages, probably resulted from viability selection favoring outcrosses, and selective removal of inbred offsprings including self-fertilized ones prior to their transplantation in the nursery. A part of this paper was orally presented at the 107th Annual Meeting of the Japanese Forestry Society (1996).     

Liquid penetration of precompressed wood VI: Anatomical characterization of pit fractures

Pit fractures of refractory coniferous heartwoods caused by precompression in the radial direction were investigated and are discussed in terms of improved liquid penetration. Small cracks appeared at the boundary between the torus and margo, along the outer margin of the margo, and on the torus when specimens were compressed and deformation was fixed by drying. The remarkable cracks were generally observed for Cryptomeria japonica D. Don. Pseudotsuga menziesii Franco showed peculiar detachment of the torus from the pit border, and Larix leptolepis Gordon exhibited only small cracks on the torus. These fractures patterns were clearer when the precompressed specimens were recovered by water impregnation and then redried.This report was presented at the 46th Annual Meeting of the Japan Wood Research Society, Kumamoto, April 1996