Isolation of protoplasts from somatic embryos of carrot

Protoplasts were isolated enzymatically from synchronously induced globular somatic embryos from a carrot suspension culture. Among the macerating enzymes tested, Driselase was the most effective for release of protoplasts from embryos. A higher medium osmolarity was required for the isolation of protoplasts from embryos than from undifferentiated cells. Protoplasts from embryos were smaller than protoplasts from undifferentiated cells. On step gradients of Ficoll, protoplasts from embryos gave one major band. Protoplasts from undifferentiated cells gave two major bands, one lighter and the other heavier than the protoplasts from embryos.     

Influence of Indigenous Microbiota on Amount of Protein and Activities of Alkaline Phosphatase and Disaccharidases in Extracts of Intestinal Mucosa in Mice

The protein content and the activities of alkaline phosphatase, maltase, and sucrase were measured at 0800, 1000, 1200, 1400, and 1600 in saline extracts of the proximal small bowels of germfree and of ex-germfree mice colonized with an indigenous microbiota. In extracts prepared from germfree mice, the total activities of all of the enzymes were relatively constant throughout the sampling period. Likewise, the total activity of alkaline phosphatase in extracts prepared from associated mice varied little as a function of time. By contrast, the total activities of maltase and sucrase in the extracts from these latter animals varied significantly from sample to sample. The total activity levels in extracts from germfree mice were approximately twofold greater than the levels in extracts from associated mice. The specific activities of alkaline phosphatase and sucrase did not vary from sample to sample in extracts prepared from either type of mouse. In contrast, the specific activity of maltase in extracts prepared from both germfree and associated mice differed significantly from sample to sample. The specific activities of all three enzymes were greater in extracts from germfree animals than in those from associated animals. The protein content of extracts prepared from germfree mice also was greater than that of extracts prepared from associated animals at every sampling time. The amount of protein extractable from the mucosa of the small bowels of the former animals varied significantly at different sampling times during the day, whereas the amount of protein extractable from the tracts of associated animals remained relatively constant throughout the day. The indigenous microbiota apparently stabilizes in some way the amount of protein extractable from the mucosa of the mouse small bowel.     

Effect of ethanol on activity of the plasma-membrane ATPase in, and accumulation of glycine by, Saccharomyces cerevisiae

The pH optimum of the ATPase activity in plasma membranes from Saccharomyces cerevisiae NCYC 431 from 8 h cultures was around 6.5 and that in membranes from organisms from 16 h cultures near 6.0. The Km[ATP] of the enzyme was virtually unaffected by the age of the culture from which organisms were harvested, although the Vmax of the enzyme in membranes from organisms from 8 h cultures was higher than that for organisms from 16 h cultures. Ethanol non-competitively inhibited ATPase activity in membranes, although the inhibition constant for the enzyme from organisms from 8 h cultures was lower than that from organisms from 16 h cultures. Glycine accumulation by the general amino acid permease was non-competitively inhibited by ethanol. Inhibition constants were virtually the same for glycine uptake by deenergized organisms from 8 h and 16 h cultures, but under energized conditions the value was greater for organisms from 16 h rather than 8 h cultures. The data indicate that inhibition of plasma-membrane ATPase activity by ethanol could account, at least in part, for inhibition of glycine accumulation by ethanol.     

Agars of Gelidiella acerosa of west and southeast coasts of India

Seventeen agar samples were extracted from Gelidiella acerosa (Forsskal) Feldmann and Hamel (Rhodophyta, Gelidiales) specimens collected from nine different sites on the Indian coast-five from southeast coast and four from the west coast. The agar samples were analysed. The stability characteristics of the gels of selected agar samples were studied by rheometry under applied stress conditions, i.e. variation of the storage (G') and loss moduli (G') were studied under varying frequency and duration (time) of the stress applied. Yield, apparent and dynamic viscosities, gelling and melting temperatures, 3,6-anhydrogalactose (3,6-AG), sulphate contents and TGA (Thermogravimetric Analysis) measurements of the products were done. It was observed that the best quality agar was produced by G. acerosa occurring in the Gulf of Mannar region in the southeast coast. The gel strengths and the viscosities of agars extracted from Gelidiella acerosa occurring in the Gulf of Mannar ranged from 500 to 700gcm(-2) and 33 to 45cP for 2001 collections and for 2002 collections the corresponding values were 450 to 845gcm(-2) and 55 to 67cP respectively. On the other hand, for the agar samples extracted from the west coast of India, the gel strength and viscosities values ranged from 225 to 400gcm(-2) and from 15 to 30cP, respectively. The agars obtained from G. acerosa collected from southeast coast have been found to be suitable for bacterial culture and molecular biology. This is the first report of superior quality of agar from the Indian agarophytes.     

Modulation of TXA2 generation of platelets by human lipoproteins

The lipoprotein (LP) fractions VLDL, LDL, HDL2 and HDL3 were prepared by ultracentrifugation of plasma from healthy volunteers and from patients with coronary heart disease (CHD). We investigated the capacity of platelets from healthy volunteers and patients with atherosclerosis to generate thromboxane A2 (TXA2) during spontaneous clotting of whole blood under the influence of the lipoprotein fractions. In our experiments the serum concentration of TXB2, reflecting the capacity of platelets to generate TXA2 during clotting, depends on several factors: the type of LP fraction used, the blood used for generation of TXA2, and for the same LP fraction whether it was taken from plasma of healthy volunteers or patients with CHD. VLDL prepared from plasma of healthy volunteers inhibited but VLDL prepared from plasma of patients with CHD enhanced the TXA2 formation of platelets from healthy volunteers (p less than 0.05, resp.). LDL from CHD patients inhibited the TXA2 formation of platelets from atherosclerotic patients (p less than 0.01). The HDL subfractions HDL2 and HDL3 from healthy volunteers inhibited TXA2 formation by platelets from healthy volunteers as well as those from atherosclerotic patients (p less than 0.05; p less than 0.01, respectively). HDL2 from patients with CHD inhibited only the TXA2 formation of platelets from healthy volunteers (p less than 0.01), whereas HDL3 from CHD patients inhibited only the TXA2 formation of platelets from atherosclerotic patients (p less than 0.01).     

Isolation of Clostridium difficile from various colonies of laboratory mice

An attempt was made to isolate Clostridium difficile from a total of 565 mice from nine different conventional mouse colonies and six different specified-pathogen-free mouse colonies. C. difficile was isolated from all the conventional colonies but from none of the specified-pathogen-free colonies. Ampicillin injected intraperitoneally increased the isolation rate of C. difficile from mouse faeces to 63.6% compared with 19.4% from untreated mice.     

Comparison of chemical analyses of boat and helicopter-collected water samples

Helicopters can be used to collect water samples from many lakes over a wide geographic area within a relatively short time period. Here we report the results from an experiment in which sequential water samples from a lake were collected first from a nonmotorized boat and then immediately afterward from a helicopter. No significant differences were found between the means of the measurement of 20 chemical parameters for the two methods of collection. When compared to obtaining samples from a boat, collection of samples from a helicopter platform had no effect on the content of the water samples.     

Studies of the GTPase domain of archaebacterial ribosomes

Ribosomes from the methanogens Methanococcus vannielii and Methanobacterium formicicum catalyse uncoupled hydrolysis of GTP in the presence of factor EF-2 from rat liver (but not factor EF-G from Escherichia coli). In this assay, and in poly(U)-dependent protein synthesis, they were sensitive to thiostrepton. In contrast, ribosomes from Sulfolobus solfataricus did not respond to factor EF-2 (or factor EF-G) but possessed endogenous GTPase activity, which was also sensitive to thiostrepton. Ribosomes from the methanogens did not support (p)ppGpp production, but did appear to possess the equivalent of protein L11, which in E. coli is normally required for guanosine polyphosphate synthesis. Protein L11 from E. coli bound well to 23S rRNA from all three archaebacteria (as did thiostrepton) and oligonucleotides protected by the protein were sequenced and compared with rRNA sequences from other sources.     

Isolation and Partial Characterization of Bacteriophages of the Phytopathogen Pseudomonas syringae

Bacteriophages isolated from culture supernatants of Pseudomonas syringae pv. syringae and from sewage were identified. The DNA from each phage was isolated and digested with the restriction endonuclease EcoRI. Eight isolates were determined to be different, with two phage isolates from sewage having restriction patterns identical to two phages from culture supernatants. The sizes of the phage DNA ranged from 24 to49 kilobases for isolates from sewage and from 39 to 52.5 kilobases for the isolates from culture supernatants. Buoyant densities of phage particles in CsCl varied from 1.498 to 1.507 g/cm³ for isolates from sewage and from 1.506 to 1.516 g/cm³ for isolates from culture supernatants. Electron microscopy revealed four morphological types. Based on plaque-forming ability of culture supernatants, 31 out of 47 strains of P. syringae are probably lysogenic.     

Additional Records of Digenetic Trematodes of Birds from Taiwan

43 species of digenetic trematodes of birds are reported from Taiwan. One new family, one new subfamily, two new genera, and nine new species are described: Taiwantrematidae fam. n. Taiwantrema arborophilae gen. et sp. n. from Arborophila crudigularis; Cyclocoelidae, Morishitium taiwanense sp.n. from A. crudigularis; Eucotylidae, Tanaisia (Tamerlania) taiwanensis sp. n. from A. crudigularis; Brachylaimidae, Leucochloridium taiwanense sp. n. from A. crudigularis; Echinostomatidae, Echinostoma taiwanense sp. n. from Zosterops japonica simplex; Dicrocoeliidae, Concinnum taiwanense sp. n. from Pycnonotus taivanus; Lecithodendriidae, Pseudocryptotropa formosae sp. n. from Pycnonotus sinensis formosae; Microphallidae, Promicrophallinae subfam. n. Promicrophallus taiwanensis gen. et sp. n. from A. crudigularis; Opisthorchiidae, Metametorchis taiwanensis sp. n. from the domestic duck.     

Formation of Dulcitol in Aerobic Dissimilation of d-Galactose by Yeasts

During the study of aerobic dissimilation of galactose by yeasts, polyhydric products were isolated in crystalline form from the fermented broths and identified. Yeast species may be divided into two groups on basis of sugar alcohol production: type I yeasts form the same end products from galactose as from glucose; type II yeasts produce dulcitol from galactose with or without other sugar alcohols but they produce no dulcitol from glucose. Isolation of dulcitol from microorganism has not been previously described.     

Chemical compositions of elastins isolated from aortas and pulmonary tissues of humans of different ages

1. Elastins were isolated from the visceral pleuras and parenchymas of lungs of humans of different ages. 2. The elastin content of pleuras increased whereas that of parenchymas remained constant with increasing age. 3. The amino acid compositions and carbohydrate contents of elastins isolated from both pulmonary tissues changed in the same way with increasing age of the subjects. These changes were similar to those observed in elastins isolated from the aorta. 4. Similar glycoproteins were isolated from pleuras and aortas, and were more difficult to extract from the elastins of older subjects. Contamination with these glycoproteins was responsible for the changes in composition of elastin, as the age of the tissue from which it was extracted increased. 5. The amount of the cross-linking amino acids desmosine and isodesmosine was lower in elastins isolated from both aorta and pulmonary tissues of senile subjects than those from younger subjects.     

Effects of serum from human subjects of various ages on proliferation of human lung and skin fibroblasts

We carried out a study to determine whether serum from old human subjects inhibited cell proliferation. The results showed that serum from old subjects of either sex did not greatly inhibit the proliferation of human fetal lung fibroblast TIG-1 cells, even when serum from subjects in their 80s was used. The same results were obtained when the effects of serum on cell proliferation were examined up to a serum concentration of 50%. It was also found that serum from old subjects did not inhibit proliferation of human skin fibroblasts from a young adult to any greater degree than serum from young adult subjects, and that serum from young adult subjects did not stimulate proliferation of skin fibroblasts from an elderly donor to any greater degree than serum from old subjects.     

Dispersal of Beauveria bassiana by the activity of nettle insects

Recent studies have shown that the entomopathogenic fungus Beauveria bassiana occurs naturally on the phylloplanes of several plants, including nettles. Insects could, by their activity, be contributing to this inoculum by dispersing it from other sites. The potential of nettle aphids Microlophium carnosum and their predator Anthocoris nemorum to disperse conidia of B. bassiana from soil to nettles and from sporulating cadavers in the nettle canopy was investigated in laboratory experiments. In petri dish assays, aphids showed potential to distribute B. bassiana from soil to nettle leaves. Predators dispersed inoculum from both soil and cadavers to nettle leaves in petri dishes. In microcosms, aphids did not disperse B. bassiana from the soil or from cadavers confined in the canopy, but A. nemorum were able to transfer inoculum from soil into the nettle canopy and to distribute conidia from cryptic cadavers. In some instances, infections were initiated in aphids and predators as a consequence of dispersal.     

New species of Choleoeimeria (Apicomplexa: Eimeriidae), coccidia of bile-bladders of reptiles, illustrating a multiplicity of host cell-parasite interrelations

Oocyst characteristics and histological features of the endogenous development of bile-bladder coccidia of the genus Choleoeimeria Paperna and Landsberg, 1989 are described and the main features for species differentiation are discussed for the following new species: C. allogamae n. sp. from Agama sp., Cameroon, West Africa; C. allogehyrae n. sp. from Gehyra australis, Magnetic Island (type) and mainland N Queensland, Australia; C. boulii n. sp. from Gehyra variegata, SW Queensland, Australia; C. calotesi n. sp. from Calotes mystaceus, Xiang-Mai, Thailand; C. heteronotis n. sp. from Heteronotia binoei, N Queensland, Australia; C. lygosomis n. sp. from Lygosoma buringi, Kon-Kaen, Thailand; C. sylvatica n. sp. from Carlia rhomboidalis, N Queensland, Australia, and C. xiangmaii n. sp. from Hemidactylus frenatus, Xiang-Mai, Thailand. Oocyst characteristic of Choleoeimeria are also reported from Oedura castelnaui, N Queensland. The described species demonstrate a diversity of associations with the bile-bladder epithelial lining, from a single parasite in a single hypertrophic host cell to multiple infections inducing the hypertrophied cells to form stratified layers, or merge into branched clumps.     

Development of plants from leaf discs of variegatedColeus and its relation to patterns of leaf chlorosis

Summary Leaf discs approximately 8 mm in diameter taken from green and from chlorotic areas of variegated leaves ofColeus were grown in light under sterile conditions in a mineral salt, sucrose, vitamin medium supplemented with auxin and cytokinin. Green shoots, which later formed roots, grew from both green and chlorotic discs in media containing suitable amounts of auxin and cytokinin. None developed in media supplemented with auxin alone or with cytokinin alone. Discs with young plants were transferred to soil. Plants that grew varied widely from those with no chlorosis to those with more chlorosis than the original variety from which the discs were taken. Plants grown from discs taken from green areas of leaves with chlorosis varied in patterns of chlorosis as much as those that grew from discs from chlorotic areas of leaves. This research was supported, in part, by The Conservation and Research Foundation.     

山羊卵母细胞的减数分裂进程

The meiotic progression of goat oocytes from follicles of different diameters was investigated in this study. The results were summarized as follows: (1) The in vitro meiotic maturation capacity was different among oocytes from follicles of different diameters. And thus oocytes from < or = 0.5 mm follicles were unable to resume meiosis; oocytes from 0.8-1.2 mm follicles were capable to resume meiosis, but could develop only to MI stage (60% at 24 h); oocytes from 1.5-5 mm follicles had acquired full-meiotic maturation capacity and 91% of them developed to M II stage at 24 h of culture. (2) The percentage of oocytes with intact-germinal vesicles from 1.5-5 mm follicles decreased significantly during 2-8 h of in vitro maturation and the decrease was even more rapid during 4-6 h of culture (from 60% to 19%, p < 0.0005). The percentage of oocytes at M I-stage increased from 24% to 61% during 6-12 h of in vitro maturation, and it then decreased. By 24 h of culture, only 2% oocytes remained at M I-stage. Twenty one percent of the oocytes in this group developed to M II-stage at 16 h of culture, and by 24 h of culture, 91% were at M II-stage. (3) Statistic analysis of the meiotic progression (the duration of each cell cycle stage) of oocytes from 1.5-5 mm follicles showed that GV stage lasted from 0 to 3 h of culture, prometaphase-I stage was from 3.0 to 7.0 h, metaphase-I stage was from 7.0 to 14.6 h, anaphase-I/telophase-I was from 14.6 to 18.4 h and metaphase-II stage lasted from 18.4 to 24 h. (4) Whether the oocytes capable of GVBD and entrance of M I developed to M II, the timing of meiotic progression prior to M I was similar. In summary, our results provided necessary data for studies on the mechanisms and control of meiosis in mammalian oocytes.     

Increasing the toxic effect of pulmonary carcinogens by destruction of epithelial-mesenchymal interaction in explants of embryonic respiratory tract of mice

The explants from 17-day old embryos of A and C57BL mice were used for long-term organ cultures. The following series of explants were studied: 1) the intact control explants; 2) the explants treated by NMU; 3) the explants treated by BP; 4) the explants isolated from mesenchyma (M); 5) the explants isolated from M and treated by NMU; 6) the explants isolated from M and treated by BP. The survival of explants treated by NMU or BP did not differ from the intact control explants (p less than 0.1). The removal of M decreased the survival only in the explants from the distal RT of A mice (p 0.01). The survival of explants isolated from M and treated by NMU or BP was significantly lower than it was in intact explants (p less than 0.01-0.001). Thus, the destruction of epithelial-mesenchymal interaction induced by removal of M can modulate the toxic effect of pulmonotropic carcinogens.     

Dependence of the structure of component parasite communities on host age

Dependence of the structure of component parasite communities on host age is studied by the example of parasite communities in minnow and grayling from the North Dvina and Pechora rivers basins. Parasite communities from immature host groups are revealed to be different from those in mature fishes by lesser number of parasite individuals and biomass, number of groups discriminated by the ratio of biomasses, and frequently by lesser number of species. Indices of diversity describing parasite communities from hosts of different ages are nearly always the same in the area examined. This observation needs in verification because values of the indices characterizing parasite communities from fishes of different ages are not equal in the parasite communities from grayling of different age from the Pinega river and minnow from the Chovju river (Dorovskikh, 2002).