Alizarin red S-stained bone and cartilage in calcium deficiency provoked by experimental liver injury in rats

Experimental liver injury with different stages was provoked in rats with daily injected doses of thioacetamide (ThAA). The dose recommended for both male and female rats was 50 mg/kg body weight. The liver damages caused were acute, subacute, cirrhotic and necrotic, with a traumatization period of 2, 7, 14 and 21 days. The loss of body weight under traumatization, indicating osteopenia, was in the case of female rats during the first experimental week markedly accelerated, and in the two subsequent weeks apparently inhibited when compared to male rats. The loss of body weight of male rats revealed a progressive fall. Vital staining was made giving intraperitoneally 200 mg/kg body weight of alizarin red S (ARS). The staining intensity was improved in the acute stage for both calvaria and tibia and in the necrotic stage for tibia only. It was impaired in the subacute stage for calvaria and tibia and in the necrotic stage for calvaria only. Prolonged traumatization with ThAA causes pathological defects in the liver and kidneys. Furthermore, the epiphyseal cartilage of necrotic-stage rats was bright red without any ARS staining.     

Acridine Orange Staining and Fluorescence of Nucleic Acids in Plasmodia and Associated Host Erythrocytes

Acridine orange in daily doses of 1, 2 and 4 mg for 4 days was given to chicks averaging 50 gm in weight. Dosage was started 1, 2 and 3 days after infection with Plasmodium gallinaceum. Such doses were sufficient to stain the parasite in vivo, as shown by its bright fluorescence in UV light, but did not exhibit any antimalarial action. Staining of fresh blood samples from infected chicks with 0.01% acridine orange in Krebs-Ringer containing 0.1 M phosphate buffer (pH 6.0-6.2) resulted in differential fluorescence of the nucleic acids of the plasmodia, to show nuclear DNA bright green and cytoplasmic RNA orange-red. After optimum acid hydrolysis, as used for the Feulgen reaction, staining with 0.1% acridine orange produced intense red fluorescence of the nuclear DNA in the plasmodia. Nuclear DNA of the chick erythrocytes showed bright fluorescence both in vivo and in vitro.     

植物细胞壁组织化学定位染色方法和技术的比较研究

利用光学和荧光显微镜比较研究几种植物细胞壁组织化学定位染色方法和技术,结果表明：（1）硫酸消化法和硫酸氢黄连素-苯胺兰对染法研究凯氏带,对取材时间和部位要求高,建议两种方法配合使用,可相互印证是否具凯氏带;（2）苏丹7B染色法,蓝色激发光下不染色和硫酸氢黄连素-苯胺兰对染研究细胞壁栓质层3种方法中,不染色蓝色激发光下结果比苏丹7B染色法敏感显色,但苏丹7B染色法在普通光学显微镜下观察较为便捷;（3）木质化细胞壁染色方法中硫酸氢黄连素-苯胺兰对染法比间苯三酚-盐酸染色法易显色观察;（4）甲苯胺兰快速染色细胞壁取代常规苏丹Ⅲ/Ⅳ法,细胞边界和层次更清楚。     

Effect of retinoids on rat spleen

It has been shown in experiments on Wistar male rats with the use of light and electron microscopy and morphometry that a single intraperitoneal injection of high doses of all-trans-methylretinoate and methyl-7,8- dihydroretinoate leads to an increase in the spleen weight, the appearance in the spleen of red cells changed in the shape, activation of phagocytic intensity of macrophages, intercellular interaction, lymphocyte proliferation and formation of new lymph follicles.     

Red Light-inhibited Mesocotyl Elongation in Maize Seedlings: II. Kinetic and Spectral Studies

Red light induces two distinct inhibition responses in mesocotyls of etiolated corn seedlings. A light dose of 10 nanoeinsteins per square centimeter is saturating for the more sensitive response, whereas doses above 1,000 nanoeinsteins per square centimeter are required to exceed the threshold sensitivity of the less sensitive one. The sensitive response can be detected within 20 minutes of the onset of illumination whereas the other response does not become apparent until more than 4 hours after the beginning of irradiation. The reciprocity law is valid for the first response, but probably not for the second. An action spectrum for the first response shows two maxima, one at 640 nanometers and the other between 660 and 670 nanometers, with a pronounced minimum near 650 nanometers. The effects both of 640 and 665 nanometers of light were reversible by far red light, but doses of far red required for full reversibility were almost three orders of magnitude greater than the doses of red required either to saturate the initial inhibition or to reverse the effect of far red light. The results suggest that corn may contain a red-absorbing pigment other than phytochrome which in some way interacts with phytochrome in the inhibition of mesocotyl elongation by red light.     

Phytochrome changes correlated to mesocotyl inhibition in etiolated Avena seedlings

The elongation of etiolated Avena mesocotyls is inhibited by red light (660 mμ). Immediately after exposing mesocotyl sections to varying doses of red light the ensuing concentrations of phytochrome in the far-red absorbing form (P₇₃₀) were measured. The extent of mesocotyl inhibition observed 5 days later is proportional to the logarithm of P₇₃₀ concentration in mesocotyl tissue at the time of red light exposure.

The inhibition of mesocotyl growth by red light can be reversed partially by subsequent exposure to far-red light (730 mμ). Increasing doses of far-red light result in decreasing concentrations of P₇₃₀ as compared with the original P₇₃₀ level due to the preceding red light exposure. The reduced mesocotyl inhibition of seedings which had been exposed to red and far-red light is proportional to the logarithm of P₇₃₀ concentration remaining in the tissue at the end of the two light exposures.

This indicates that the same correlation exists between P₇₃₀ concentration and growth response whether the seedlings had been exposed to red light only or to red followed by far-red light.

     

Morphological alterations and NO-synthase expression in the heart after continuous light exposure of rats

Although exposure to continuous light is associated with hypertension and modulates the outcome of ischemia-reperfusion injury, less attention has been paid to its effects on cardiac morphology. We investigated whether 4-week exposure of experimental rats to continuous 24 h/day light can modify cardiac morphology, with focus on heart weight, fibrosis and collagen I/III ratio in correlation with NO-synthase expression. Two groups of male adult Wistar rats were studied: controls exposed to normal light/dark cycle (12 h/day light, 12 h/day dark) and rats exposed to continuous light. After 4 weeks of treatment the absolute and the relative heart weights were determined and myocardial fibrosis and collagen type I/III ratio were evaluated using picrosirius red staining. Endothelial and inducible NO-synthase expression was detected immunohistochemically. The exposure of rats to continuous light resulted in an increase of body weight with proportionally increased heart weight. Myocardial fibrosis remained unaffected but collagen I/III ratio increased. Neither endothelial nor inducible NO-synthase expression was altered in light-exposed rats. We conclude that the loss of structural homogeneity of the myocardium in favor of collagen type I might increase myocardial stiffness and contribute to functional alterations after continuous light exposure.     

Dose-response curves for radish seedling phototropism

Radish seedlings (Raphanus sativus L.) were grown for 4 days in complete darkness, or in white light, or for 3 days in darkness followed by 1 day of red light. Phototropic dose-response curves for the seedlings grown in these three ways were determined with 460-nm light. The dark-grown and red light-treated seedlings responded with positive curvatures of no more than 10° to energy doses in the first positive range and with larger positive curvatures in the second positive dose range. No indifferent or negative curvature was seen with the light intensity used. White light-grown seedlings did not respond to first positive energy doses, but responded as strongly to second positive doses as the other types of seedlings.     

Effects of photodynamic treatment on biological antioxidant systems in rats

Effects of photodynamic treatments on inherent antioxidant metabolites and cellular defence enzymes have been investigated in rats. Wistar rats were grouped into untreated controls, light controls, hematoporphyrin derivative (Hpd) (treated with 5 and 10 mg Hpd/kg body weight and kept in dark) and sets treated with both Hpd and red light (dose 172 and 344 j/m2 ). After 2, 24, 48 and 72 hr of Hpd injection the rats sacrificed, livers quickly excised to analyze Hpd uptake, activities of enzymes like catalase, GSH-Px and antioxidants like GSH, vitamin A, vitamin E and vitamin C. The results showed that the loss of Hpd from liver as a function of post- injection time was non- linear. An increased generation of lipid radicals was observed in the groups treated with 5 mg Hpd and higher dose of light and in groups treated with 10 mg Hpd at both the doses of light. Combination of light and Hpd reduced hepatic GSH content with a concomitant reduction in GSH-Px. At higher doses of Hpd and light, there was a significant reduction in hepatic vitamin A levels. Combination of Hpd and light in all doses reduced vitamin E content in liver. The decreased biological antioxidant contents and GSH-Px may be attributed to their utilization for the scavenging of free radicals generated by Hpd and light in tissues. However, no change in catalase activity and vitamin C content in liver was noted in experimental rats. The results suggest that exposure to higher doses of Hpd with light alters oxidant stress system and TBARS content in rat.     

Localization of expression of KNAT3, a class 2 knotted1-like gene

KNAT3 is a class 2 kn1 -like gene in Arabidopsis thaliana . The RNA expression patterns of KNAT3 were characterized through the use of promoter-GUS fusion analysis and in situ hybridization. KNAT3 is expressed in several tissues and at several times during development. There are three main expression patterns: (1) during early organ development in young leaves, buds and pedicels; (2) at and near the junction between two organs at specific times during development, including the hypocotyl-root boundary in young seedlings, the anther-filament junction in mature flowers, and the ovule-funiculus and peduncle-silique boundaries in elongating siliques; and (3) in maturing tissues such as the style of elongating siliques, the petioles of maturing leaves, and most of the root. The varied expression patterns may indicate that KNAT3 plays several different roles in plants, depending on when and where it is expressed. Previous work on KNAT3 (Serikawa et al. , 1996) indicated that expression of its RNA is regulated by light. Promoter-GUS seedlings were grown under different light conditions (continuous white, red and far-red light) to examine more closely the light regulation of the KNAT3 promoter. Continuous white light resulted in stronger overall GUS staining in the same patterns seen in seedlings grown under long-day conditions (cotyledons, upper hypocotyl and roots). Continuous red light resulted in reduced GUS expression in those same tissues. Continuous far-red light led to seedlings showing stronger staining in the hypocotyl and cotyledons than red light-grown plants but no staining in the roots. Thus, the KNAT3 promoter responds differently to red and far-red light.     

水稻原生质体细胞核及原生质体融合体的简易染色观察法

筛选出一种荧光染料罗丹明B(Rhodamine B),利用该荧光染料染色,原生质体细胞核在普通光学显微镜或荧光显微镜下呈红色或发出强烈的桔红色荧光,能清晰地进行分辨。利用罗丹明B或者使用荧光染料FDA,对两种不同来源的原生质体进行染色,在荧光显微镜下,两种不同来源的原生质体分别发出桔红色或绿色荧光,因此可以用于原生质体融合中不同的融合体类型的观察和分析。     

Influence of light colours on growth and stress response of pearl gourami <Emphasis Type="Italic">Trichopodus leerii</Emphasis> under laboratory conditions

This study aimed to investigate the influence of illumination with different light spectra on growth and stress response in pearl gourami Trichopodus leerii Bleeker, 1852. Fish (6.35 ± 0.43 cm, 5.69 ± 0.67 g) were reared in 8 glass aquaria each with 25 fish under one of four different lighting conditions: yellow (max 546 nm), red (max 605 nm), blue (max 470 nm) and white (control). Experiments lasted for 90 days. The stress response was evaluated by measuring cortisol levels. No significant effect was observed on final body weight, length and specific growth rate of fish. However, best values for these characteristics were achieved for fish reared under yellow light. A significant higher weight gain, condition factor was found in fish reared under yellow light compared with fish reared under blue or red light. In contrast, a lower final body weight, specific growth rate and weight gain were achieved for fish reared under red light. Feed conversion ratio was significantly lower in fish under yellow light compared with red or blue light. Stressed fish showed lower cortisol levels under yellow light compared with other light exposures. The study indicates that, pearl gourami grows better under yellow light and that yellow light lowers the stress-induced cortisol response.     

Reproducibility of morphometric measurements of amyloid after various staining methods

Four histologic staining methods used for detecting amyloid (Congo red, viewed in both normal and polarized light, Sirius red, Crystal violet and Thioflavine T) were applied to heart muscle autopsy samples from 19 patients who suffered from amyloidosis. The amount of amyloid present was evaluated with morphometry (point counting) by five pathologists, and the interobserver reproducibility and variation of point counting in these staining methods were analyzed. The Sirius red method showed the least variation and was the most suitable stain for demonstrating amyloid with respect to reproducibility. Thioflavine T showed the greatest variation and was the least suitable stain with respect to reproducibility. The range of variation was considerable in all staining methods. The results show that stains differ in their specificity and sensitivity in staining amyloid, observers differ in their interpretation of staining results and certain stains result in more uniform interpretations than do others.     

Persistence of red light induction in lettuce seeds of varying hydration

Incubation of lettuce seeds (Lactuca sativa L. cv. Grand Rapids) in 0.3 m mannitol allows sufficient water uptake to make seeds fully sensitive to red light. But germination is possible only after lowering the osmotic potential of the incubation medium. The red light induction of these incompletely hydrated seeds can be reversed by far red light. Their reversibility declines with time at a slower rate than seeds incubated in water. About half the seeds in 0.3 m mannitol respond to far red light when all seeds in water have escaped control by far red light. Close to 100% of the seeds remain sensitive to far red exposure if 0.6 m mannitol is used as osmoticum. The retention of the original red light stimulus is inversely related to the concentration of the incubation medium.The fresh weight of viable seeds incubated with water or with an osmoticum increases rapidly during the first 5 hours, then remains stationary for about 12 hours. After that only germinating seeds experience a second increase in fresh weight. Heat-killed seeds do not show such a discontinuity in water uptake.     

A Simple Method Using Alizarin Red S For the Detection of Calcium in Epoxy Resin Embedded Tissue

This report presents a simple procedure for staining 1-2 μm epoxy plastic sections of cells and mineralizing matrix present in fetal bovine bone tissue cultures. A 0.3% aqueous toluidine blue 0 solution was used as a cellular stain and was followed with 2% alizarin red S for the detection of calcium at sites of mineralition. Effects of concentration and pH of alizarin red S on the penetration of epon embedded thick sections were investigated Optimal staining was achieved with a 2% aqueous alizarin red S solution adjusted to a pH of 5.5-6.5. This staining procedure provides unusually clear contrast between mineral and bone cells in plastic sections for light microscopy.     

Effect of different light qualities on the ultrastructure, thylakoid membrane composition and assimilation metabolism of Chlorella fusca

Chlorella fusca (Shihira et Krauss) strain C-1.1.10 was grown under three different light qualities (red, white or blue light) in homocontinuous cultures. Under electron microscopy, blue light cultures showed enlarged cells, thinner cell walls and lower starch content than red light cells. Under blue light, the degree of stacking of the thylakoid membranes was significantly lower than under white or red light conditions. Changing the light from blue to red the ratio of exposed to appressed membranes was doubled. Compared to red light cells, blue light cells exhibited higher photosynthetic rates per chlorophyll molecule and contained less chlorophyll per dry weight. Blue light stimulated the content of soluble protein as well as that of soluble carbohydrates. The dry weight productivity per unit time was enhanced under blue light conditions. The thylakoid protein complexes which are generally assumed to be localized in the exposed membranes were found in higher concentrations under blue light than under red light. In blue light, both the Photosystem II/Photosystem I ratio and the ratio of light-harvesting chlorophyll protein to P-700 chlorophyll a -protein were lower than in red light. Blue light cells contained twice the concentration of cytochrome f , which correlates well with their higher photosynthetic capacity. When altering the light quality, the degree of change in the reaction center complexes was much lower than expected given the corresponding degree of change in the ratio of exposed to appressed membranes. These results are discussed in light of the question as to whether the variation in the stoichiometry of the laterally distributed complexes can be explained by changes in the degree of stacking alone.     

Sequential Use of Wright's and Ziehl-Neelsen's Stains for Demonstrating Phagocytosis of Bacterial Spores

Nongerminating spores, germinating spores, and vegetative cells of Clostridium botulinum type A were observed during phagocytosis in the peritoneal fluid of white mice. Since phagocytes are easily ruptured by heat, the method described avoids heating, as this has been employed in conventional spore staining methods. A thin smear of the fluid is air dried on the slide for 2 hr, and stained by Wright's method: stain, 2 min; dilution water, 2 min; and rinsed; then in 0.005% methylene blue for 30 sec, and rinsed. This is followed by Ziehl-Neelsen's stain for 3-4 min and destained with 1: acetone-95% ethanol for 10 sec. The slide is rinsed, and Wright's staining repeated: stain 1 min, dilution 2-3 min; and thereafter washed about 5 ml of Wright's buffer. Blotting and air drying completes the staining. Non-germinating spores stain light red with a red spore wall, germinating spores are deep red throughout, vegetative cells are blue, and leucocytes show a dark purple nucleus and light blue cytoplasm.     

Light Dependence of Chloroplast Replication and Starch Metabolism in the Moss Polytrichum commune

Spores of Polytrichum conwtuine were grown on a mineral salt solution with or without sucrose and exposed to continuous white light, continuous darkness, red light and/or far-red light. With sucrose, germination and filament growth occurred in all conditions, Without sucrose, germination and filament growth occurred only in light. Two phytochrome mediated responses of the chloroplasts were demonstrated. Chloroplast replication occurred in continuous white light and red light of 15 min/6 hours. In continuous darkness and in far red light of 15 min/6 hours, the size of the chloroplasts increased; but no replication occurred. Both the chloroplast replication and chloroplast size were red, far-red light reversible. When changed from one continuous light environment to another, a lag period occurred before the chloroplasts responded to the new environment. Electron micrographs of sections and in vivo staining of the chloroplasts with iodine solution demonstrated that the change in size of the chloroplasts was at least partially due to the synthesis and degradation of starch.     

Phytochrome-mediated Germination Responses in gamma-Irradiated Lettuce Seeds

Sublethal doses of γ-radiation and far red light have some-what analogous, red light reversible, effects on the germination of lettuce seeds (Lactuca sativa L. var. Grand Rapids). However, the mechanism by which γ-radiation retards germination appears to differ from that of far red light. Compared to controls, γ-radiation retarded germination for the first 24 hours; but after 36 or 48 hours of imbibition gemination of treated seeds was higher than that of the controls, whether or not the γ-irradiated seeds received red or far red light. The effects of γ-radiation are more pronounced in seeds containing 15% water at the time of treatment than in those containing only 7% water. The promotive action of red light is operative in the presumed absence of cell division in γ-treated seeds.     

The Effect of Red Irradiation on Plastid Ribosomal RNA Synthesis in Dark-grown Pea Seedlings

Dark-grown pea seedlings (Pisum sativum L.) were irradiated for a short period each day with low intensity red light (662 nm), red light immediately followed by far red light (730 nm), or far red light alone. Other plants were transferred to a white light regime (14 hours light/10 hours dark). There was no change in the amount of RNA in the tissue on a fresh weight basis after the various treatments. However, compared with dark-grown seedlings, those plants irradiated with red light showed an increase in the net RNA content per stem apex. In addition there was a two- to three-fold increase in ribosomal RNA of the etioplasts relative to the total ribosomal RNA. These increases were comparable to those found in plants grown in the white light regime. The changes were much smaller if the dark-grown plants were irradiated either with red light followed by far red light, or with far red light alone. Thus continuous light is not essential for the production of ribosomal RNA in plastids, and the levels of ribosomal RNA found in chloroplasts can also be attained in etioplasts of pea leaves in the dark provided the leaf phytochrome is maintained in its active form.