Isolation of xanthomegnin from Penicillium viridicatum by preparative high-pressure liquid chromatography

A method was developed for the production and purification of xanthomegnin from Penicillium viridicatum (NRRL 6430) cultured on rice at 15 degrees C for 29 days. Liquid-liquid extraction followed by high-pressure liquid chromatography afforded 440 mg of crystalline xanthomegnin per kg of rice.     

Evaluation of five green fluorescence-emitting streptavidin-conjugated fluorochromes for use in immunofluorescence microscopy

 Fluorescein isothiocyanate (FITC) is largely used in immunofluorescence methods. We propose to analyse the quality of some recent fluorochromes using image analysis. Fluorochromes tested include FITC and dichlorotriazinylaminofluorescein (DTAF), dipyrrometheneboron difluoride (BODIPY), Rhodol Green and cyanine 2. RAMOS cells were immunolabelled against the proliferating cell nuclear antigen (PCNA) revealed by the biotin-streptavidin technique. Slides were mounted in anhydrous glycerol or in buffered glycerol (pH 7.0 or pH 8.5). No antifading medium was added. Cell fluorescence emission intensity and bleaching characteristics were measured. Rhodol Green exhibited the highest fluorescence intensity and the best photobleaching resistance. Although BODIPY also resisted well during the photobleaching assay, its fluorescence intensity was weak. FITC, DTAF and cyanine 2 showed intermediate fluorescence intensity and a fast decay of fluorescence. Among the green emitting fluorochromes tested, Rhodol Green appeared to be the best. Accepted: 1 April 1996     

Is there an accumulation of cells during G2 in some acute lymphoblastic leukaemias?

In some cases of acute lymphoblastic leukaemia (ALL) the percentage of cells in G2 + M is higher than anticipated when compared with the percentage in S phase. This increase in G2 + M, as detected by flow cytometry measurement of DNA content, may be due to an accumulation of cells, either in G2 or during the end of S phase; it may also be related to the existence of small tetraploid clones generally ignored by cytogeneticists. In order to identify possible subpopulations of cells with a DNA index greater than or equal to 2.0, we have compared the results of a cytogenetic analysis to the G2 + M values. We have also studied the distribution of S phase cells in 24 cases of ALL by incorporating 5-bromodeoxyuridine, labelling the cells by indirect immunofluorescence, and analysing them by flow cytometry after propidium iodide staining. The distribution of cells during S phase was quantified: no accumulation of cells was ever observed at the end of S phase. The question of the existence of small tetraploid clones, G2 arrested cells or cells with a G2 elongation remains open. However, we feel that it is more probable that, in this pathology, an elongation of the duration of G2 occurs.     

The Conservation of Saproxylic Insects in Tropical Forests: A Research Agenda

Invariably, insects are overlooked when tropical forest management issues are discussed, because there are so many species, they are taxonomically intractable and so poorly known. Often people take the view that if you look after the vegetation and vertebrates, the insects will look after themselves. This may be true for some functional groups, but for saproxylic insects, this seems unlikely. Their study deserves high priority, since they are dependent on the very resource – wood – whose removal from the ecosystem is the usual object of forest management. Given the current international effort to develop 'criteria and indicators' to monitor sustainable forest management for biodiversity values, there is a window of opportunity for sound ecological research on saproxylic insects to influence the formulation of forest policy such that their needs can be taken into account. There is already a large body of knowledge on temperate and boreal region saproxylic insects, and on the effects that logging has on them, but knowledge of the tropical forest situation lags far behind. This paper proposes a research agenda to enable the needs of saproxylic insects to be taken into account in natural forest management in the tropics. Basic questions, such as whether logging has so far had an impact on tropical saproxylic insects, and whether there are workable sampling techniques to investigate this, still remain to be addressed and deserve high priority. The links between the responses of saproxylic insects and more 'charismatic' study species need to be investigated. We also need to know whether there is a correlation between the intensity of logging and the response of saproxylic insects, and, critically, whether we would be justified in measuring some surrogate aspects of forest structure (as potential habitat for saproxylic insects) rather than the saproxylic insects themselves, and modelling this to determine likely impacts of different management regimes. We consider such an ambitious research agenda as justified given the scale of impact that forest use and management is likely to have on tropical forest insects in the future.     

The ABC of a Simple Method for Pulmonary Vein Angiography

Catheter-directed intervention to treat atrial fibrillation (AF) is becoming widely accepted procedure in current clinical practice. For assessment of pulmonary vein (PV) anatomy, angiography of left atrium (LA) and/or PV is often performed. We present a new, simple angiographic method for PVs and LA opacification using SL1 sheath. Total of 100 patients in our clinic underwent this procedure. In all of the cases good angiographic results were achieved. No immediate or late complications related to this procedure were observed.     

Ancylostoma ceylanicum: immunization with soluble worm extract and responses to challenge infection of dogs

When dogs were immunized with soluble extract of adult Ancylostoma ceylanicum antigen, they were partially resistant to challenge infection in this model of human hookworm infection. Two immunizing doses, each of 1 mg protein suspended in Freund's complete adjuvant, were administered to one group of animals 1 and 3 weeks prior to infection with 5000 larvae. When compared with control dogs given the same infective dose, fecal egg excretion and intestinal adult worm burden in the immunized animals were reduced by 59 and 74%, respectively. Infection had no significant effect on hemoglobin concentrations, mean red cell volumes, total white cell counts, platelet levels, or spontaneous and phytohemagglutinin-induced lymphocyte transformations in both control and immunized animals. Both groups developed an eosinophilia, and lymphocytes from the immunized dogs responded transiently to stimulation with both larval and adult worm antigens. Specific IgM antibodies were transitory in both groups of dogs following infection. IgG antibodies developed significantly 2 weeks after infection in the immunized group; however, they did not appear until 4 weeks after infection in the control group. Both groups developed IgA antibodies 1 week after infection. They were maintained in the control dogs, in contrast to the levels in immunized animals which subsided rapidly 4 weeks after infection. Therefore, when animals are injected with soluble adult worm antigen prior to infection, specific protective immunity is acquired.     

Rapid method for the assay of 4-aminobutyric acid (GABA), glutamic acid and aspartic acid in brain tissue and subcellular fractions

The thin-layer electrophoretic separation at pH 4.8 of brain extracts and a procedure for fluorescent staining of the plates with fluorescamine are described for the rapid routine determination of 4-aminobutyric acid (GABA), glutamic acid and aspartic acid in brain extracts and in particulate fractions of brain tissue. Automated sample application, electrophoretic separation using two chambers, and quantitation by in situ fluorescence scanning allows the assay of 280 samples within three working days. The method is reproducible (S.D. <8% of the mean) within the range of 0.2–2 nmole per spot. The staining procedure can be applied to a variety of related analytical problems. The method has proved useful for the determination of the specific radioactivities of GABA, glutamic acid and aspartic acid in metabolic studies.