Comparative observation of freeze–thaw-induced damage in pig, rabbit, and human corneal stroma

Objective  Although variations exist between species with respect to outcomes after cryopreservation, little is known about the differences in the susceptibility of the corneal stroma to cryoinjury. We performed this study to investigate freeze–thaw-induced damage in keratocytes and collagen in rabbit, pig, and human corneas.
Animals studied  Rabbit, pig, and human.
Procedures  We prepared 250-μm-thick anterior stroma from rabbit, pig, and human corneas after scraping off the epithelium and endothelium. Each 250-μm-thick corneal stroma without epithelium was placed in a 50-mL tube, frozen with liquid N₂ for 15 min and taken out to thaw rapidly at 37 °C. This procedure of rapid freezing and thawing was repeated three times. Differences between the species with respect to cells and collagen structures were examined using hematoxylin–eosin (H&E) staining, terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay, and transmission electron microscopy (TEM). We orthotopically transplanted the pig and rabbit corneal transplants after the triple freeze–thaw cycle into rabbit eyes and evaluated graft survival.
Results  On gross examination, rabbit corneas became opaque after the triple freeze–thaw procedure, while pig and human corneas remained transparent. Histologically, keratocytes were apoptotic on TUNEL assay and TEM in rabbit, pig, and human corneas. Collagen fibrils were fragmented and the arrangement of collagen fibrils was severely disturbed in rabbit corneas on H&E staining and TEM; collagen was well preserved in pig and human corneas. Rabbit corneal stroma underwent autolysis after transplantation, whereas the pig corneal stroma remained clear for 1 month.
Conclusions  Our study showed that rabbit corneal stroma was more susceptible to freeze–thaw injury than pig and human corneas.     

Growth Responses and Differences in Gene Expression depending on Cultivation Temperature between Alternative type Wheat Varieties

Journal of Crop Science and Biotechnology - Two domestic wheat varieties were grown in the growth chamber set at 17°C, 20°C, 23°C and 26°C on average after the emergence of...     

水稻中抗品种抗褐稻虱机制的研究

褐稻虱在中抗水稻品种上的刺探频率低于抗级品种。虽然褐稻虱在中抗品种上能正常取食,但同化利用率较低。中抗品种受害后,无论是植株损害系数、光合作用强度降低程度或产量损失率均显著较感虫品种的少,说明了受害后较之感虫品种具有较强的补偿能力。另外,水稻品种受害后呼吸强度均较健株的明显降低。     

Measurement of Atmospheric Formaldehyde and Monoaromatic Hydrocarbons using Differential Optical Absorption Spectroscopy during Winter and Summer Intensive Periods in Seoul, Korea

Ozone effects on Mediterranean forests were studied for 10 years in southern France. The aim of these studies was to understand the impact of the pollutant on foliar damage among Aleppo pine (Pinus halepensis Mill) and Arolla pine (Pinus cembra L) populations. During the summers of 2001 and 2002, ozone concentrations were measured using UV absorption analysers (4) and passive samplers (29) at two forest monitoring sites: Site 1 in the Nice coastal area for Aleppo pine and Site 2 in the Mercantour mountains for Arolla pine, in the vicinity of tree observation sites. AOT 40 values were quite high (25–33 ppm.h), especially in the Mercantour mountains, for the 5 months of measurement. Foliar damage (defoliation and discoloration) and specific symptoms (chlorotic mottle) were observed on 8 Aleppo pine and 13 Arolla pine plots. For Arolla pines, there was a significant correlation between defoliation and chlorotic mottles. There was also a correlation between mean ozone concentrations measured by passive samplers and chlorotic mottles. These correlations were less significant for Aleppo pine.     

A new Brucella canis species-specific PCR assay for the diagnosis of canine brucellosis

Brucellosis is a zoonotic disease that is transmitted from animals to humans, and the development of a rapid, accurate, and widely available identification method is essential for diagnosing this disease. In this study, we developed a new Brucella canis species-specific (BcSS) PCR assay and evaluated its specificity and sensitivity. A specific PCR primer set was designed based on the BCAN_B0548-0549 region in chromosome II of B. canis. The PCR detection for B. canis included amplification of a 300-bp product that is, not found on other Brucella species or, genetically or serologically related bacteria. The detection limit of BcSS-PCR assay was 6 pg/μl by DNA dilution, or 3 × 10³ colony-forming units (CFU) in the buffy coats separated from whole blood experimentally inoculated with B. canis. Using the buffy coat in this PCR assay resulted in approximately 100-times higher sensitivity for B. canis as compared to detect directly from whole blood. This is the first report of a species-specific PCR assay to detect B. canis, and the new assay will provide a valuable tool for the diagnosis of B. canis infection.     

Methylation methods for the quantitative analysis of conjugated linoleic acid (CLA) isomers in various lipid samples

Precise methylation methods for various chemical forms of conjugated linoleic acid (CLA), which minimize the formation of t,t isomers and allylmethoxy derivatives (AMD) with the completion of methylation, were developed using a 50 mg lipid sample, 3 mL of 1.0 N H(2)SO(4)/methanol, and/or 3 mL of 20% tetramethylguanidine (TMG)/methanol solution(s). Free CLA (FCLA) was methylated with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). CLA esterified in safflower oil (CLA-SO) was methylated with 20% TMG/methanol (100 degrees C, 5 min), whereas CLA esterified in phospholipid (CLA-PL) was methylated with 20% TMG/methanol (100 degrees C, 10 min), followed by an additional reaction with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). Similarly, CLA esterified in egg yolk lipid (CLA-EYL) was methylated by base hydrolysis, followed by reaction with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). These results suggest that for the quantitative analysis of CLA in lipid samples by GC, proper methylation methods should be chosen on the basis of the chemical forms of CLA in samples.     

Cell proliferation and neuroblast differentiation in the dentate gyrus of high-fat diet-fed mice are increased after rosiglitazone treatment

In this study, we determined how rosiglitazone (RSG) differentially affected hippocampal neurogenesis in mice fed a low-fat diet (LFD) or high-fat diet (HFD; 60% fat). LFD and HFD were given to the mice for 8 weeks. Four weeks after initiating the LFD and HFD feeding, vehicle or RSG was administered orally once a day to both groups of mice. We measured cell proliferation and neuroblast differentiation in the subgranular zone of the dentate gyrus using Ki67 and doublecortin (DCX), respectively, as markers. In addition, we monitored the effects of RSG on the levels of DCX and brain-derived neurotrophic factor (BDNF) in hippocampal homogenates. At 8 weeks after the LFD feeding, the numbers of Ki67- and DCX-positive cells as well as hippocampal levels of DCX and BDNF were significantly decreased in the RSG-treated group compared to the vehicle-treated animals. In contrast, the numbers of Ki67- and DCX-positive cells along with hippocampal levels of DCX and BDNF in the HFD fed mice were significantly increased in the RSG-treated mice compared to the vehicle-treated group. Our data demonstrate that RSG can modulate the levels of BDNF, which could play a pivotal role in cell proliferation and neuroblast differentiation in the hippocampal dentate gyrus.     

Photoperiod and Soil Munition Constituent Effects on Phytoaccumulation and Rhizosphere Interactions in Boreal Vegetation

Permafrost thaw is expected to alter biogeochemistry and hydrology, potentially increasing the mobility of soil constituents. Northern latitude boreal forests where permafrost thaw is occurring also experience extreme changes in day length during the growing season. As the effects of photoperiod on plant uptake of soil constituents or interactions with the rhizosphere are unknown, our objective was to determine these interactions with three plant species from different functional groups. A tree, forb, and grass common to military training ranges in this region were grown in soil spiked with or without lead, antimony, or 2,4-dinitrotoluene and grown under 16, 20, or 24 h of light. Plant biomass, soil constituent uptake, and rhizosphere bacterial communities were compared between treatments. Photoperiod had no effect on plant uptake of any soil constituent or on rhizosphere community, indicating that plants and their associated microbial communities adapted to this environment are resilient to extremes in photoperiod. Lead uptake was not significant in any plant species and had no effect on the rhizosphere. Antimony increased the percentage composition of Saprospirales in the rhizospheres of two of the three plants, indicating an interaction between this bacterial order and antimony. Antimony uptake by white spruce (Picea glauca) was considerable, with a mean concentration of 1731 mg kg^?1 in roots, while mean shoot concentration was only 155 mg kg^?1, indicating its potential to phytostabilize this heavy metal. Although antimony had the strongest impact on the rhizosphere bacterial community, it was also readily accumulated by the grass and tree.     

Synthesis and biological evaluation of [1-(1H-1,2,4-triazol-1-yl)alkyl]-1-silacyclopentanes

A series of 1-aryl-1-(1H-1,2,4-triazol-1-yl)alkyl-1-silacyclopentanes has been synthesized by four-step reactions starting from 1-chloroalkyltrichlorosilane and tested for fungicidal activities in vitro for ten fungi and in vivo for four fungi occurring in rice, cucumber, tomato etc. Biological activities of the title compounds are strongly dependent upon the p-substituent on the phenyl group in the following order: F>Cl>Ph>OEt>H. Especially, 1-p-fluorophenyl-1-[1-(1H-1,2,4-triazol-1-yl)alkyl]-1-silacyclopentanes (alkyl=methyl or ethyl) showed significant fungicidal activity with a broad spectrum comparable to flusilazole in in-vivo assay. © 1998 SCI.