Inhibition of human lymphocyte activation by wheat germ agglutinin: a model for saccharide-specific suppressor factors

The suppressive effect of wheat germ agglutinin (WGA) on lectin-stimulated blastogenesis and immunoglobulin production was studied. Addition of WGA at 10 micrograms/ml inhibited phytohemagglutinin (PHA)-, concanavalin-A (Con-A)-, and pokeweed mitogen (PWM)-induced mitogenic responses by 70-80%. PWM-driven immunoglobulin synthesis was suppressed by 45% with WGA. The inhibitory effects of WGA were not due to cell death or to interference with lectin binding at the cell surface. Inhibition was dependent on the presence of WGA in the cell culture during the first 24 hr of mitogen exposure and was observed in cultures of both monocyte-depleted peripheral blood mononuclear cells as well as T-cell-enriched populations. WGA-induced inhibition of blastogenesis was blocked by the addition of N-acetylglucosamine (GluNAc) which prevents WGA binding to the cell surface. WGA was found to mimic the suppressive effect of a soluble immune suppressor supernatant (SISS) derived from Con-A-activated mononuclear cell cultures. PHA responses were inhibited by 80 and 95% with SISS and WGA, respectively. The inhibition by both WGA and SISS was totally reversed with addition of GluNAc. Furthermore, WGA and SISS demonstrated competition for the same cell surface receptor site. WGA may therefore be useful as an in vitro model of a saccharide-specific, biologically relevant, soluble mediator for the investigation of mechanisms of immunologic suppression.     

Fermentation of a milk-soymilk and Lycium chinense Miller mixture using a new isolate of Lactobacillus paracasei subsp. paracasei NTU101 and Bifidobacterium longum

A milk–soymilk mixture was fermented using Lactobacillus paracasei subsp. paracasei NTU101 and Bifidobacterium longum BCRC11847 at different inoculum ratios (1:1, 1:2, 1:5, 2:1, and 5:1). When the inoculum ratio was 1:2, the cell numbers of both strains were balanced after 12 h of cultivation. The pH and titratable acidity were very similar at the various inoculum ratios of cultivation. The milk–soymilk mixture was supplemented with 5, 10, 15, and 20% Lycium chinense Miller juice and fermented with Lactobacillus paracasei subsp. paracasei NTU101 and B. longum BCRC11847. Sensory evaluation results showed that supplementation with 5% Lycium chinense Miller juice improved the acceptability of the fermented milk–soymilk. The fermented beverage was stored at 4°C for 14 days; variations in pH and titratable acidity were slight. The cell numbers of L. paracasei subsp. paracasei NTU101 and B. longum BCRC11847 in the fermented beverage were maintained at 1.2×10⁹ CFU/ml and 6.3×10⁸ CFU/ml, respectively, after 14 days of storage.     

Intra-tissue Characteristics of Chloroplasts in the Lamina and Petiole of Mature Winter Leaf of Arum italicum Miller

Abstract: Laminae and petioles from mature winter leaves of Arum italicum were studied in order to obtain information on the sun—shade intra-tissue properties of chloroplasts. This inference was based on the: (1) micro- and submicroscopic characteristics of the chloroplasts, (2) cytochemical localizations of functional PS I and PS II, (3) pigment patterns and compositions, (4) immunolocalization of Rubisco, and (5) net photosynthesis. It was inferred that all the chloroplasts across the lamina had adaptations to intermediate shade conditions, without a sun-shade dimorphism between the palisade and the spongy tissues. In the petiole, where normally-structured chloroplasts were surprisingly present in the entire thickness of the organ, a structural and chemical dimorphism was found between the outer chlorenchyma and the inner aerenchyma where intermediate shade-type and extreme shade-type chloroplasts were present, respectively. However, some anomalies in the pigment composition were noted chiefly in the inner aerenchyma (low concentrations of β-carotene and lutein, absence of zeaxanthin, presence of unusual pigments, for instance lutein epoxide, lutein cis-isomer, and chlorophyllide a). The Rubisco immunolabelling in the outer chlorenchyma of the petiole was similar to that in the lamina, while it was very scant in the inner aerenchyma. Net photosynthesis in the petiole was about 75% of that recorded in the lamina. These data suggest that the petiole of the mature winter leaf of A. italicum closely co-operates with the lamina for enhancing light capture and utilization.     

Mutagenesis and mutation transfer induced by ultraviolet light in plasmid-cloned DNA

We describe here simple techniques for increasing the frequency of UV-induced mutations in a DNA fragment cloned in plasmid pBR322. Irradiation of both the host and the plasmid DNA before transformation is necessary to produce new mutations in the plasmid DNA, presumably because the UV-damaged pBR322 replicon cannot efficiently induce the error-prone repair pathway of Escherichia coli. In contrast, U V irradiation of the plasmid DNA alone before transformation primarily causes the transfer of preexisting mutations from the host chromosome to homologous DNA present in the plasmid. The only other kind of mutants obtained were large deletions of the plasmid DNA. Two chromosomal mutations from the host galK gene and one from the lacZ gene have been transferred to the plasmid by UV irradiation of the plasmid DNA alone. The technique can thus be of general use.     

Edward Hitchcock’s Pre-Darwinian (1840) “Tree of Life”

The “tree of life” iconography, representing the history of life, dates from at least the latter half of the 18th century, but evolution as the mechanism providing this bifurcating history of life did not appear until the early 19th century. There was also a shift from the straight line, scala naturae view of change in nature to a more bifurcating or tree-like view. Throughout the 19th century authors presented tree-like diagrams, some regarding the Deity as the mechanism of change while others argued for evolution. Straight-line or anagenetic evolution and bifurcating or cladogenetic evolution are known in biology today, but are often misrepresented in popular culture, especially with anagenesis being confounded with scala naturae. Although well known in the mid 19th century, the geologist Edward Hitchcock has been forgotten as an early, if not the first author to publish a paleontologically based “tree of life” beginning in 1840 in the first edition of his popular general geology text Elementary Geology. At least 31 editions were published and those between 1840 and 1859 had this “paleontological chart” showing two trees, one for fossil and living plants and another for animals set within a context of geological time. Although the chart did not vary in later editions, the text explaining the chart did change to reflect newer ideas in paleontology and geology. Whereas Lamarck, Chambers, Bronn, Darwin, and Haeckel saw some form of transmutation as the mechanism that created their “trees of life,” Hitchcock, like his contemporaries Agassiz and Miller, who also produced “trees of life,” saw a deity as the agent of change. Through each edition of his book Hitchcock denounced the newer transmutationist hypotheses of Lamarck, then Chambers, and finally Darwin in an 1860 edition that no longer presented his tree-like “paleontological chart.”     

Phenylalanine promotes interaction of transmembrane domains via GxxxG motifs

Interactions of transmembrane helices play a crucial role in the folding and oligomerisation of integral membrane proteins. In order to uncover novel sequence motifs mediating these interactions, we randomised one face of a transmembrane helix with a set of non-polar or moderately polar amino acids. Those sequences capable of self-interaction upon integration into bacterial inner membranes were selected by means of the ToxR/POSSYCCAT system. A comparison between low/medium-affinity and high-affinity sequences reveals that high-affinity sequences are strongly enriched in phenylalanine residues that are frequently observed at the − 3 position of GxxxG motifs, thus yielding FxxGxxxG motifs. Mutation of Phe or GxxxG in selected sequences significantly reduces self-interaction of the transmembrane domains without affecting their efficiency of membrane integration. Conversely, grafting FxxGxxxG onto unrelated transmembrane domains strongly enhances their interaction. Further, we find that FxxGxxxG is significantly over-represented in transmembrane domains of bitopic membrane proteins. The same motif contributes to self-interaction of the vesicular stomatitis virus G protein transmembrane domain. We conclude that Phe stabilises membrane-spanning GxxxG motifs. This is one example of how the role of certain side-chains in helix-helix interfaces is modulated by sequence context.     

Use of bioclimatic indexes to characterize phenological phases of apple varieties in Northern Italy

The research was designed to characterize the phenological behaviour of different apple varieties and to compare different bioclimatic indexes in order to evaluate their adaptability in describing the phenological phases of fruit species. A field study on the requirement for chilling units (winter chilling requirement) and the accumulation of growing degree hours of 15 native apple cultivars was carried out in a fruit-growing area in North West Italy (Cuneo Province, Piedmont). From 1991 to 1993, climatic data were collected at meteorological stations installed in an experimental orchard (Verzuolo, Cuneo). Four methods were compared to determine the winter chilling requirement: Hutchins, Weinberger-Eggert, Utah and North Carolina. The Utah method was applied to determine the time when the chilling units accumulated become effective in meeting the rest requirements. A comparison of the different methods indicated that the Weinberger-Eggert method is the best: as it showed the lowest statistical variability during the 3 years of observations. The growing degree hour requirement (GDH) was estimated by the North Carolina method with two different base temperatures: 4.4°C and 6.1°C. More difficulties were met when the date of rest completion and the beginning of GDH accumulation was determined. The best base temperature for the estimation of GDH is 4.4°C. Phenological and climatic characterizations are two basic tools for giving farmers and agricultural advisors important information about which varieties to choose and which are the best and the most correct cultivation practices to follow. Received: 25 October 2000 / Revised: 6 August 2001 / Accepted: 6 August 2001     

Quadruple colocalization of calretinin,calcitonin gene-related peptide,vasoactive intestinal peptide,and substance P in fibers within the villi of the rat intestine

Double-labeling immunofluoresenct histochemistry demonstrates that calretinin, a calcium-binding protein, coexists with calcitonin gene-related peptide, vasoactive intestinal peptide, and substance P in the fibers innervating the lamina propria of the rat intestinal villi. An acetylcholinesterase histochemical stain revealed that the majority of calretinin-containing cells in the myenteric ganglia were cholinergic and that about one half of the submucosal calretinin-containing cells colocalized with acetylcholinesterase. In situ hybridization studies confirmed the presence of calretinin mRNA in the dorsal root ganglia, and a ribonuclease protection assay verified the presence of calretinin message in the intestine. The coexistence of calretinin in calcitonin-gene-related-peptide-containing cells that also contained substance P and vasoactive intestinal polypeptide in the dorsal root ganglia suggest that these ganglia are the source of the quadruple colocalization within the sensory fibers of the villi. Although the function of calretinin in these nerves is unknown, it is hypothesized that the coexistence of three potent vasodilatory peptides influences the uptake of metabolized food products within the vasculature of the villi.