Aniline Blue Research Articles

Hybridization between hexaploid triticale and Triticum monococcum L. I. Progamic reaction

One triticale (AUAUBBRR) was crossed with 15 forms of Triticum monococcum (AbAb) from different geographical regions. Twenty-four hours after hand pollination, the ovules were removed from the florets and fixed and stained in a solution containing water-soluble aniline blue. By means of a fluorescent microscope, assessments were made for the amount of pollen on stigmatic surface, the value of pollen germination, and the length of pollen tubes, calculated in diameters of the corresponding pollen grain. An absolute progamic barrier was observed in none of the 15 initial combinations. Five to 20 percent of pollen grains germinated, with important differences between the pollinators. The latter was observed for pollen tube length, too. A contingency table analysis revealed a strong correlation between both traits. It was shown that quantitative differences of this kind can only be detected if the real data distribution is used. A transformation into vectors or ranks, was less informative. Furthermore, a correlation between the quantitative differences mentioned above and the number of fertilized ovules was visible. In case of pollen germination and pollen tube length, (–values were 0.69*and 0.71*, respectively. It can be concluded that there is some kind of “specific crossability” even in such complicated distant hybridization. It is necessary, therefore, to involve more than one T. monococcum form, independent of a more or less uniform habit of this material.

エナメル・象牙境の観察―脱灰標本において象牙質表面に出現する網目状構造物について―

As part of morphological research to investigate the development and function of the human dentino-enamel junction, structural features of net-like structures on the decalcified dentin surface of upper and lower incisors were observed by using stereoscopic microscope, and scanning and transmission electron microscopes. The results were as follows : 1) The net-like structures on the decalcified dentin surface were stained by hematoxylin, orange G phosphomolybdic acid, toluidine blue and aniline blue. 2) The majority of net-like structures were covered with a membrane-like structure. 3) An individual reticulation of net-like structures corresponds to the margin of each dentin foveolus. 4) The dentin crystals situated in the vicinity of the dentinoenamel junction were larger than the crystals of usual dentin and were more closely aligned. On the other hand, the enamel crystals in this region were smaller than those in other layers of enamel. The surface dentin in the vicinity of the dentino-enamel junction seemed to be highly calcified. 5) It was possible to clearly demonstrate the outline and distribution of dentin foveolus by using certain dyes on the decalcified specimen. 6) By observing the distribution and structural features of net-like structures in detail by applying this dye method to human and animal teeth, the formation mechanism of the dentino-enamel junction and functions of those teeth were elucidated morphologically.

Evaluation of sperm chromatin integrity and seminal quality in HIV positive patients

To identify the effect of HIV infection on DNA maturation process and integrity of sperm cells, as well as the degree of alteration in seminal parameters using aniline blue and acridine orange tests. Prospective, descriptive, comparative study. Twenty HIV positive male patients were included in the case group. Sperm chromatin was evaluated with acridine orange and aniline blue tests. Spermogram, hepatitis B, hepatitis C, Chlamydia and syphilis detection test were performed in every patient. Results were compared with an equal size group of healthy controls. Statistical evaluation was done using t test, simple correlation and logistic regression. There was no significant difference in aniline blue and acridine orange tests results in cases group compared with controls (P>0.05). Mean test results in HIV positive patients for aniline blue was 79.1% (±22.2), for acridine orange was 52.2% (±23.3). A statistically significant direct correlation was observed between aniline blue test result and acrinidine orange test result, sperm concentration, vitality and morphology in HIV patients (Figure 1). A direct correlation was observed between acridine orange test result and sperm concentration, semen volume, vitality and motility in the same group (Figure 2). There was no correlation between HIV viral load or CD4 count with chromatin integrity test results or spermogram parameters alterations. There is no difference in sperm chromatin alterations was observed in HIV positive patients compared with healthy controls. There is no correlation of the degree of sperm chromatin or semen parameters alterations with HIV viral load or CD4 count in HIV positive patients. Analine blue and orange acridine tests results are correlated with most semen parameters results in HIV patients.

Effect of sperm chromatin integrity on the embryo quality following ICSI

DNA cytometry could differentiate damaged (sub-haploid) sperm from mature condensed haploid sperm, while aniline blue (AB) test indicates deficient DNA protamination. AB test can differentiate between healthy (non-stained) and non-healthy (stained) sperm chromatin. The aim of our study was to evaluate and compare sperm chromatin status using these two techniques with the embryo quality in infertile couples undergoing ICSI for male factor infertility. Prospective study. Semen samples from 42 infertile men with no female factor infertility. Routine semen analysis was done according to the WHO guidelines (1999). An aliquot of semen was used for ICSI procedure. Embryos were scored for morphological appearance. Pregnancy test was done 2 week after the embryos transfer. Sperm quality was assessed by DNA cytometry and Aniline blue staining. Good quality embryos showed significantly lower percentage of sub-haploid sperm compared to poor embryo quality group (P value = 0.02) (see Table).TableComparison of sperm chromatin structure in good vs. poor quality embryos∗P<0.05 considered significant by the Student's t test. Open table in a new tab ∗P<0.05 considered significant by the Student's t test. Embryo quality after fertilization in ICSI can be affected by the sperm chromatin fragmentation. Aniline blue staining is unable to differentiate the sperm that will result in good equality embryos. Selecting sperm with higher sperm chromatin integrity will help in the improvement of embryo quality after ICSI.

Preweaning exposure to enriched environment induces hippocampal neurogenesis: experiment with rats

To explore the effects of preweaning exposure to enriched environment on hippocampal neurogenesis and the underlying mechanisms. Thirty-six 10-day-old SD rats were randomly divided into the 2 equal groups: control group and enriched environment group (EE group. From the age of 10 days to 24 days the rats received intraperitoneal injection of bromodeoxyuridine (BrdU) 50 mg/kg every other day to label the newly proliferated cells in vivo, and the rats in EE group were daily exposed to enriched environment for 20 minutes. Six rats of each group were sacrificed whren they were 24 days of age. Nuclear protein of the hippocampus was extracted to undergo Western blotting to detect the levels of calmodulin and phosphorylated CREB (cAMP response element binding). Other rats were sacrificed at the age of 63 days. Coronal cryostat sections of brain were cut. Sections at the level 3.6 mm posterior to the bregma were obtained and stained with methyl aniline blue and the number of cells in the hippocampal dentate gurus (DG) of the right hemisphere were counted using x 400 microscope. BrdU immunohistochemistry and double immunofluorescence labeling with BrdU/NeuN or BrdU/GFAP were done, and the numbers of BrdU-labeled cells and ratios of neurons and astrocytes differentiated from BrdU-labeled cells were calculated. The levels of calmodulin and phosphorylated CREB in the hippocampal nuclear extract of the EE group were 0.065 +/- 0.035 and 0.485 +/- 0.007 respectively, both significantly higher than those of the control group (0.245 +/- 0.035 and 0.220 +/- 0.014 respectively, P = 0.01 and P = 0.002). The number of cells in the DG area of right hippocampus 3.6 mm posterior to bregma of the EE group was 1580 +/- 72, significantly higher than that of the control rats (1375 +/- 62, t = -7.461, P < 0.01). The number of BrdU labeled cells of the EE group was 5363 +/- 487, significantly higher than that of the control group (2984 +/- 318, t = -14.177, P < 0.01). The ratio of neurons of the EE group was 85.0% +/- 2.8%, significantly higher than that of the control group (80.2% +/- 2.8%, t = -4.166, P < 0.01). The differentiation rate of astrocytes of the EE group was 4.0% +/- 0.5%, significantly higher than that of the control group (2.6% +/- 0.6%, t = -6.493, P < 0.01). Preweaning exposure to enriched environment can induce neurogenesis. The underlying mechanism may be that enriched environment induces the activation of calmodulin and CREB in hippocampus.

Assessment of the Possibility of Direct Crossing Between Cultivated Potato and Two Wild Allotetraploid Relatives

The cultivated potato Solanum tuberosum L. is an autotetraploid with Endosperm Balance Number (EBN) of 4. There are some allotetraploid 2EBN wild species with desirable traits for potato improvement. However, it is very difficult to cross between these two groups due to differences in EBN. Eleven genotypes of autotetraploid cultivated potato including the six cultivars of subsp. Tuberosum (tbr) and five clones of subsp. Andigena (adg) were crossed with two wild allotetraploid species: S. acaule (acl) and S. stoloniferum (sto). For asses the pollen-pistil incompatibility, some ofpollinated flowers were fixed in Carnoy's solution and stained with aniline blue. Evaluation with fluorescent microscope revealed that pre-zygotic pollen-pistil incompatibility in these crosses is frequent. The pistils of both subsp. of cultivated potato were incompatible with pollens of acl but when sto used as staminate parent it was compatible with five cultivars of subsp. Tuberosum, but incompatible with most ofsubsp. Andigena (4 clones). From 739, 4x, 4EBN x 4x, 2EBN crosses we couldn't get any viable seed. In reciprocal crosses, allotetraploid species were pollinated with a mix of pollens from Tuberosum or Andigena clones. The crosses of sto x adg, acl x adg and acl x tbr were compatible but in the case of sto x tbr the situation was not clear. From these crosses 157 fruits were obtained but most of them contained only shrink seed and we find just one hybrid from the sto x tbr combination. The hybrid was tetraploid (2n = 4x = 48) and vigorous with profuse flowering and good fertility. When this hybrid was used as the pistillate parent it was compatible with both parental species.

Involvement of arabinogalactan proteins in the regeneration process of cultured protoplasts of Marchantia polymorpha

Protoplasts of Marchantia polymorpha L. (liverwort) regenerated new cell walls in initial culture. However, the survival rate of regenerated cells decreased rapidly after this stage. The decrease in survival rate was suppressed by the β‐glucosyl Yariv reagent (βglcY), which binds to arabinogalactan proteins (AGPs), only when it was added to culture medium during the period of incipient cell wall regeneration. The addition of βglcY after the period of incipient cell wall regeneration had no effect on the survival rate. These results suggested the involvement of AGPs in the cell wall regeneration process. After cell wall regeneration, the regenerated cells started to divide actively after being transferred to a medium with 1% activated charcoal (AC). Protoplasts that had been cultured with βglcY during the period of incipient cell wall regeneration and then transferred to the AC medium divided vigorously, and the cell division rate was remarkably increased (&gt;80%). However, without transfer to the AC medium, βglcY at concentrations higher than 20 μg ml−1 inhibited cell division. No effect on cell survival nor cell division was observed with the α‐galactosyl Yariv reagent. Staining of β‐1,3‐glucan (callose) with aniline blue (AB) showed that a large amount of β‐1,3‐glucan was deposited in the regenerated cell walls of the protoplasts cultured without βglcY, while little or no β‐1,3‐glucan was stained by AB in protoplasts cultured with βglcY. These results suggest that AGPs and β‐1,3‐glucan play important roles in the survival and subsequent cell division of regenerated cells of M.polymorpha protoplast cultures.

Assessment of nuclear DNA integrity of epididymal spermatozoa following experimental chronic spinal cord injury in the rat

Infertility is considered as one of the major problems associated with spinal cord injury (SCI). However, the exact underlying mechanism is still unknown. Therefore, the main objective of this experimental study was to evaluate the effect of chronic SCI on sperm parameters as well as chromatin integrity and DNA of spermatozoa aspirated from cauda epididymis of rats. Forty-five adult Wistar rats were divided into three groups - SCI, sham, and control. Following laminectomy, SCI was induced onto exposed dura matter (T10). The sham group underwent laminectomy of T10 only, while the control rats were not exposed to any type of injury or medication. The cauda epididymal sperms were aspirated after 8 weeks for analysis of sperm parameters and sperm chromatin integrity with aniline blue (AB), chromomycin A3 (CMA3), sodium dodecyl sulphate (SDS), and acridine orange (AO) tests. The sperm progressive motility and normal morphology of SCI rats were significantly changed when compared with other groups (p < 0.05). In addition, AB as well as CMA3 tests were insignificantly increased in the SCI group when compared with the sham and control groups. However, SDS and AO tests were significantly changed in SCI samples when compared with the sham and control groups (p < 0.001). The results showed that chronic SCI in rat disturbs sperm parameters as well as nuclear maturity and DNA integrity of sperms. Therefore, sperm chromatin structure is compromised in SCI animals as revealed by chromatin structural probes. These alterations may reduce the fertility potential of the male gamete following SCI.

Luminescence of aniline blue in hydrophobic cavity of BSA

The fluorescence of aniline blue in the presence of bovine serum albumin (BSA) was investigated using total luminescence spectroscopy (TLS), fluorescence emission and synchronous fluorescence spectra. The fluorescence exhibits appreciable hypsochromic shift along with an enhancement in the fluorescence yield. It is suggested that the polarity-dependent twisted intramolecular charge transfer (TICT) process is responsible for the remarkable fluorescence sensitivity. The reduction in the rate of TICT in the hydrophobic interior of albumin leads to the increase of fluorescence yield. In BSA little rotation around N–C bond of aniline blue occurs during the excited state lifetime. Therefore, it could be concluded that aniline blue molecules have been included into the hydrophobic cavity of BSA and formed complex. The driving forces behind such association between the dye and biopolymer might include electrostatic attraction between ionic dye species and opposite charges on the biopolymer. In the synchronous fluorescence spectra of aniline blue when Δ λ = 170 nm, dual fluorescence for aniline blue is observed, local excited state (LE) and TICT state, the relative fraction of the normal planar state increased quickly in low BSA concentration. It is indicated that hydrophobic pockets of BSA affected the LE-TICT equilibrium.

Effects of Single Intratracheal Exposure to Chlorhexidine Gluconate on the Rat Lung

Chlorhexidine gluconate (CHX) is an antiseptic that has been widely used for disinfection of cutaneous wound and gingivae. Recently, a patient who inhaled CHX solution died from acute respiratory distress syndrome (ARDS). Although it is highly possible that direct pulmonary damage might be the cause of ARDS, there is no preclinical information about the pulmonary toxicity of CHX. In the current study, the acute direct action of CHX to the lung was evaluated in rats. We successfully exposed the left but not the right lung either to CHX at concentrations of 1%, 0.1%, and 0.01% or to saline using a curved-tip administration tube. At the higher concentrations of CHX (0.1% and 1%), severe congestion to the alveoli and capillaries and perivascular and intra-alveolar hemorrhages were observed 1 day after exposure. Aniline blue–stained collagen fibers with an infiltration of inflammatory cells were present 7 days after exposure. The fibrotic changes and intra-alveolar inflammatory cells had decreased but were still observed sporadically 28 and 84 days after exposure. These detrimental effects were more severe at 1% than at 0.1% CHX. No remarkable effect was observed after exposures to 0.01% CHX and saline. We were able to evaluate the time-course changes in the pulmonary toxicity of CHX by exposures limited to the left lung. It is highly possible that CHX at a concentration of more than 0.1% might directly induce ARDS when aspirated and reaching to the alveoli

An improved method for clearing and staining free-hand sections and whole-mount samples.

Free-hand sectioning of living plant tissues allows fast microscopic observation of internal structures. The aim of this study was to improve the quality of preparations from roots with suberized cell walls. A whole-mount procedure that enables visualization of exo- and endodermal cells along the root axis was also established. Free-hand sections were cleared with lactic acid saturated with chloral hydrate, and observed with or without post-staining in toluidine blue O or aniline blue. Both white light and UV light were used for observation. Lactic acid was also used as a solvent for berberine, and fluorol yellow for clearing and staining the samples used for suberin observation. This procedure was also applied to whole-mount roots with suberized celllayers. Clearing of sections results in good image quality to observe the tissue structure and cell walls compared with non-cleared sections. The use of lactic acid as a solvent for fluorol yellow proved superior to previously used solvents such as polyethylene glycol-glycerol. Clearing and fluorescence staining of thin roots such as those of Arabidopsis thaliana were successful for suberin visualization in endodermal cells within whole-mount roots. For thicker roots, such as those of maize, sorghum or tea, this procedure could be used for visualizing the exodermis in a longitudinal view. Clearing and staining of peeled maize root segments enabled observation of endodermal cell walls. The clearing procedure using lactic acid improves the quality of images from free-hand sections and clearings. This method enhances the study of plant root anatomy, in particular the histological development and changes of cell walls, when used in combination with fluorescence microscopy.

Morphology after synergistic terminal end-to-side nerve graft repair: Investigation in a nonhuman primate model

BACKGROUND: Functional efficacy of synergistic terminal end-to-side nerve graft repair in a median nerve defect model in nonhuman primates was recently published elsewhere. The purpose of this study was to investigate the morphology of the end-to-side repair site and the nerve graft used after synergistic terminal end-to-side nerve graft repair. METHODS: Experiments were carried out in seven adult baboons. Synergistic end-to-side nerve graft repair bridging from the terminal motor branch of deep branch of the ulnar nerve to the thenar motor branch of the median nerve was performed after transection of the median nerve at forearm level. Specimens were harvested three months after surgery. Chromotrope aniline blue was used for staining neural collagenic connective tissue. S100-antibody was used to quantify the number of Schwann cells. Axon counts were performed using NF (neurofilament) antibody staining. To enable profound morphological assessment, specimens were harvested 5 mm proximal to the end-to-side nerve graft repair site (deep branch of ulnar nerve), at the end-to-side nerve graft repair site (proximal graft), and at the distal end-to-end nerve graft suture site. The contralateral terminal deep branch of ulnar nerve served as a control. RESULTS: The area of collagenic connective tissue was 6% larger at the end-to-side repair site and 28% larger at the distal end of the nerve graft. Highest mean values were found at the distal site of the nerve graft. This was due to very strong collagenization of the nerve graft of animal number six. In other words, there was a moderate enhancement of collagenic connective tissue within the nerve graft in six animals. Schwann cell counts showed similar values within all nerve segments. There was no difference when compared to nonoperated control. The number of axons distal to the end-to-side repair site was smaller by 19% on average. Additionally, we found an average 23% decrease of axons at the end-to-end coaptation site compared to data of the deep branch of ulnar nerve. CONCLUSIONS: The results in this morphological study of synergistic end-to-side nerve graft repair at the level of very peripheral terminal motor branches in a nonhuman primate model demonstrate effective distal axonal sprouting and successful Schwann cell survival. As expected, we found a modest increase of neural collagenic connective tissue within the nerve graft in most animals (6 out of 7 animals).

Class 1 Reversibly Glycosylated Polypeptides Are Plasmodesmal-Associated Proteins Delivered to Plasmodesmata via the Golgi Apparatus

SE-WAP41, a salt-extractable 41-kD wall-associated protein that is associated with walls of etiolated maize (Zea mays) seedlings and is recognized by an antiserum previously reported to label plasmodesmata and the Golgi, was cloned, sequenced, and found to be a class 1 reversibly glycosylated polypeptide ((C1)RGP). Protein gel blot analysis of cell fractions with an antiserum against recombinant SE-WAP41 showed it to be enriched in the wall fraction. RNA gel blot analysis along the mesocotyl developmental axis and during deetiolation demonstrates that high SE-WAP41 transcript levels correlate spatially and temporally with primary and secondary plasmodesmata (Pd) formation. All four of the Arabidopsis thaliana (C1)RGP proteins, when fused to green fluorescent protein (GFP) and transiently expressed in tobacco (Nicotiana tabacum) epidermal cells, display fluorescence patterns indicating they are Golgi- and plasmodesmal-associated proteins. Localization to the Golgi apparatus was verified by colocalization of transiently expressed AtRGP2 fused to cyan fluorescence protein together with a known Golgi marker, Golgi Nucleotide Sugar Transporter 1 fused to yellow fluorescent protein (GONST1:YFP). In transgenic tobacco, AtRGP2:GFP fluorescence is punctate, is present only in contact walls between cells, and colocalizes with aniline blue-stained callose present around Pd. In plasmolyzed cells, AtRGP2:GFP remains wall embedded, whereas GONST1:YFP cannot be found embedded in cell walls. This result implies that the targeting to Pd is not due to a default pathway for Golgi-localized fusion proteins but is specific to (C1)RGPs. Treatment with the Golgi disrupting drug Brefeldin A inhibits Pd labeling by AtRGP2:GFP. Integrating these data, we conclude that (C1)RGPs are plasmodesmal-associated proteins delivered to plasmodesmata via the Golgi apparatus.

Characteristics of Photocatalytic Titania Fabricated by Anodization

The anodic titanium dioxide (TiO2) films for photocatalyst were prepared by anodization in acid solution. The characteristics of surface layer and the photocatalytic efficiency of the anodic film have been investigated. The chemical states of anodic film were analyzed using XPS, and the photocatalytic efficiency was evaluated by the degradation rate of aniline blue. From XPS results the component elements of the electrolyte, P and S, were observed in the anodic film, which were incorporated from the electrolyte into the oxide layer during anodization. The porous TiO2 film exhibited high photocatalytic efficiency for degradation of aniline blue.

Persistent histones in immature sperm are associated with DNA fragmentation and affect paternal contribution of sperm: A study of aniline blue staining, fluorescence in situ hybridization (FISH) and DNA nick translation

We have tested whether aniline blue staining of sperm is related to DNA chain breakage. During sperm development there are concurrent nuclear and cytoplasmic processes, including meiosis, DNA repair, histone-protamine replacement, and cytoplasmic extrusion. Aniline blue, a marker of persistent histones in immature sperm, may stain sperm light (L, mature sperm), intermediate (IN) and dark (D, immature sperm, high level of persistent histones). It was reported that the immature D sperm do not show FISH chromosome labeling, whereas in L and IN there are well characterized FISH signals. We have examined, with a combination of aniline blue staining and FISH or aniline blue staining and DNA nick translation, whether the lack of FISH signals in D sperm is caused by an interference of aniline blue staining with FISH, or if in immature D sperm there is also DNA fragmentation, thus the FISH probes fail to recognize the chromosomes. Sperm were stained with aniline blue, and the microscopic fields of aniline blue stained sperm were digitized and captured by the computer assisted Metamorph program (Universal Imaging Co., PA). Subsequently, the aniline blue stain was bleached with 30% methanol-DDW, and the slides were subjected to either multicolor FISH using X, Y, and 17 chromosome probes, or to DNA nick translation. Nick translation generates color via enzyme linked nucleotides inserted at DNA chain breaks. The color intensity is proportional with the extent of DNA fragmentation, and graded as light nick (LN), intermediate nick (IMN) and dark nick (DN) sperm. Further, the saved aniline blue stained sperm images were compared with those of the same sperm after FISH or DNA nick translation treatment. Statistical evaluation by Student t, Mann-Whitney, and the Spearman’s correlation tests was carried out by the SigmaStat program (Jandel, CA). We confirmed that D sperm indeed fail to show FISH signal, whereas in L and IN sperm the fluorescence chromosomes are well detectable. The sperm stained for aniline blue and nick translation provided evidence for extensive DNA degradation in D sperm. In 5 men we studied 600 spermatozoa. There was an agreement between the staining patterns with aniline blue and subsequent nick translation in 82.8% of sperm (500 of 600): 46.5% of sperm stained as L and LN, 27.7% of sperm stained IM and IMN, and 7.6% of sperm stained D and DN, respectively. Thus, sperm with high level of persistent histones have also shown extensive DNA degradation. Only 10 sperm of 600 had an L-DN or D-LN discordant pattern. The correlation between the aniline blue and nick translation staining patterns was r=0.82, p<0.001. Using probes for persistent histones and fragmented DNA in the same sperm indicates that diminished maturity sperm show defects in both histone-protamine replacement and DNA fragmentation. Both persistent histones and DNA fragmentation affect the paternal contributions of sperm to the zygote. Thus, aniline blue staining may facilitate the quick and inexpensive prospective evaluation of DNA integrity in husbands treated with assisted reproduction.

The Use of Aniline Blue for the Determination of Dead Phytoplankton, Zooplankton and Meroplankton in LC50 Testings After 96 Hours… A Re-Evaluation of the US Environmental Protection Agency Methodology

There is a need for a reliable staining technique to distinguish between live and dead organisms following LC50 tests. This is especially so in cases where organisms can be stressed or even become unconscious and appear dead to the aided or naked eyes. Visual observations under such conditions can result in an LC50 value shifting to the lower concentration thereby imposing stiffer guidelines for compliance. Aniline blue can only stain individuals which are physiologically dead imposing an accurate live-dead evaluation and producing a true LC50 value. Guidelines imposed using such data will facilitate compliance and provide an accurate value for an LC50.

TiO2계 나노 복합촉매 특성에 미치는 생성상의 영향

In the present study, <TEX>$TiO_2$</TEX> imbedded composite powders have been successfully prepared from the (Cu. Zn)/<TEX>$TiO_2$</TEX> composite salt solution. The composite (Cu, Zn)/<TEX>$TiO_2$</TEX> powders were formed by drying the solution at 200~<TEX>$600^{\circ}C$</TEX> in the hydrogen atmosphere. Photocatalytic characteristics was evaluated by detecting the decomposition ratio of aniline blue with UV-visible spectrophotometer(Shimazu Co., UV-1601). Phase analysis of (Cu, Zn)/<TEX>$TiO_2$</TEX> composite powders was carried out by XRD and DSC, and powder size was measured with TEM. The mean particle size of composite powders was about 100mm. As the reduction temperature increases, a few zinc sulfide and oxide phases was formed and copper oxide phase was reduced. The decomposition ratio of aniline blue was about 80％ under the UV irradiation by the TiO<TEX>$_2$</TEX> phase in the composite (Cu, Zn)/<TEX>$TiO_2$</TEX> powders and similar decomposition ratio of 80％ was obtained at the UV lightless condition by virtue of Cu and Zn compounds.

Removal of textile dyes from aqueous solutions by adsorption on biodegradable wastes

The adsorption of the textile dyes Basic Blue 41, Aniline Blue, Reactive Black 5 and Mariposa Blue was studied from model solutions prepared in distilled water in the concentration range up to 2000 mg 1–1. Biodegradable wastes were used and are proposed here as adsorbents due to their abundance and inexpensiveness in the authors’ country. The influence of the particle size, pH, presence of an electrolyte in the dye solution, adsorbent activation and competition in ternary dyes mixtures was determined in this study. Adsorption capacity of the adsorbents designated as HOMAP and POMAP with particle diameter less than 3 mm for the Basic Blue 41 dye was found to be 154 and 77 mg g‐1, respectively. The efficiency of the prepared adsorbents for removing basic, acid and reactive dyes with a concentration of 400 mg 1–1 was examined. A comparison of the adsorption capacity and desorption feasibility of the adsorbents was made. The basic dye was adsorbed with efficiencies varying from 87–98%, the reactive dye was removed in the range 60–98% and the add dye, 60–70% by three of the four adsorbents tested in this study. The determination of adsorption equilibrium concentrations was followed by HPLC with diode array detection.

Development of novel elongated fiber-structure in protoplast cultures of Betula platyphylia and Larix leptolepis

Novel elongated fiber-structures were repeatedly found both in leaf protoplast culture of two clones of Betula platyphylla and in protoplast culture of embryogenic cells of Larix leptolepis. Suboptimum culture conditions for cell division appeared to lead to fiber formation when using multi-well plate culture with varying medium compositions The suboptimum conditions for cell divisions were brought about by (1) plant growth regulators: auxins and cytokinins; (2) pH: 3.5, 4.5, 5.8; (3) divalent cations: CaCl2 and MgCl2; and (4) sugars: sucrose and mannitol. Divalent cations had the most profound effect on fiber formation. Calcium ions were preferred by Betula and magnesium ions were preferred by Larix. Single fiberpurification and micro-staining methods using a micromanipulator were developed. The fibers fluoresced when stained with Calcofluor White and Aniline Blue, which suggested that they were composed of cell wall component(s), including callose (β-1,3-glucan). Electron microscopy showed that fiber bundles of Larix fibers had helical substructures.

New insight into the mechanism of cellulose and callose biosynthesis: proteases may regulate callose biosynthesis upon wounding

Using a silver-enhanced, gold-secondary antibody immuno-location approach, we investigated the mechanisms for the switch from β-1,4- to β-1,3-glucan biosynthesis upon wounding. Antibodies against β-1,4- and β-1,3-glucan synthases were used to locate these synthases before and after wounding of Mung bean (Vigna radiata var Berken) hypocotyls. Within 5 min of wounding, β-1,4-glucan synthases which were densely localized on plasma membranes adjacent to the secondary walls at the wound site completely disappeared, and β-1,3-glucan synthases became labeled. The immuno-location of the β-1,3-glucan synthases in the secondary walls was in good accordance with the region where the β-1,4-glucan synthases were localized before wounding. Aniline blue was also utilized to visualize the deposition of callose upon wounding. Within 5 min of wounding, callose had accumulated in the corresponding region where the immuno-labeling of β-1,3-glucan synthase was detected after wounding. The β-1,3-glucan synthases were always detected from the sieve plate and plasmodesmata which are known to have constitutive synthesis of callose regardless of wounding. Secondary walls located distantly into the tissue away from the wound site were consistently labeled by the β-1,4-glucan synthase antibody even after wounding. Immuno-blot analysis clearly shows that the levels of β-1,4-glucan synthase subunit Ces A decreased dramatically within 30 min, whereas the β-1,3-glucan synthase subunit CFL1 levels increased significantly after wounding. The intensity of labeling reached a maximum at the wound site, and gradually decreased in correspondence with the distance from the wound site. When a protease inhibitor cocktail was applied upon wounding, neither the β-1,3-glucan synthase appeared nor callose was deposited during the first 5 min of wounding. On the other hand, β-1,4-glucan synthase was detected at the wound site, implying that activation of β-1,3-glucan synthase may rely on the degradation of the β-1,4-glucan synthase. Our study may provide new insight into β-glucan synthesis in higher plants.