Development of immunoassays for human interleukin 3 and interleukin 4, some of which discriminate between different recombinant DNA-derived molecules

Two panels of hybridomas were produced that secreted monoclonal antibodies (MAbs) against recombinant DNA-derived human interleukin 3 and interleukin 4 (rhIL-3 and rhIL-4). From each panel, sensitive immunoradiometric assays (IRMAs) were developed which were capable of detecting the recombinant molecule used as the immunogen but were unable to recognize natural or other recombinant forms of the same cytokine. Subsequent studies using the MAbs from each panel showed that a number of the MAbs appeared only to recognize that particular recombinant molecule used as immunogen, with little or no binding to other recombinant forms of the molecule. By using MAbs that were found to be unrestricted in their recognition for different recombinant forms of the cytokines, it was possible to develop an IRMA for IL-4 that was capable of detecting natural IL-4 as well as all the recombinant forms equally. An IRMA was also developed for IL-3 but was not of equivalent sensitivity in detecting the different recombinant forms of IL-3 used in the study. The recombinant DNA-derived cytokine molecules used to raise the two panels of MAbs contained amino acid substitutions relative to the natural sequences, and these findings indicate that caution should be exercised when using immunoassays to estimate natural sequence molecules if antibodies raised to modified rDNA-derived molecules are used.     

Functional homogeneity of the non-mitochondrial Ca2+ pool in intact mouse lacrimal acinar cells.

In the absence of extracellular Ca2+, treatment of mouse lacrimal acinar cells with maximal concentrations of methacholine released Ca2+ from intracellular stores. No additional Ca2+ was mobilized by subsequent application of the intracellular Ca(2+)-ATPase inhibitor, thapsigargin, the stable inositol 1,4,5-trisphosphate ((1,4,5)IP3) analog, inositol 2,4,5-trisphosphate ((2,4,5)IP3) (by microinjection), or the Ca2+ ionophore, ionomycin. However, following prolonged activation of cells by methacholine in the presence of extracellular Ca2+, Ca2+ accumulated into a pool which was released by ionomycin but not by thapsigargin. This latter accumulation was blocked by prior microinjection of ruthenium red, indicating that it represents mitochondrial uptake. In saponin-permeabilized lacrimal cells, two Ca(2+)-sequestering pools were detected: (i) a ruthenium red-sensitive, thapsigargin-insensitive pool, presumed to be the mitochondria; and (ii) a ruthenium red-insensitive, thapsigargin-sensitive pool. Only the thapsigargin-sensitive pool accumulated Ca2+ at concentrations similar to those in unstimulated cells. The thapsigargin-sensitive Ca2+ pool was sensitive to (1,4,5)IP3; however, in contrast to findings in intact cells, only 44% of this pool was releasable by (1,4,5)IP3 or (2,4,5)IP3. These data indicate that, in intact lacrimal acinar cells, all exchangeable (ionomycin-sensitive) Ca2+ residues in a pool which responds homogeneously to agonists, (1,4,5)IP3, and thapsigargin. Prolonged elevation of [Ca2+]i results in Ca2+ accumulation into a second, ruthenium red-sensitive pool, presumably mitochondria. Finally, permeabilization of the cells fragments the non-mitochondrial pool, resulting in two pools, one sensitive and one insensitive to (1,4,5)IP3.     

Biology of Furcellaria lumbricalis (Hudson) Lamouroux (Rhodophyta: Gigartinales), a commercial carrageenophyte

Information on the commercially utilized marine red alga Furcellaria lumbricalis is summarized from published sources. Biological aspects under discussion include: nomenclature, taxonomy, morphology, development and life history, distribution, phenology, ecophysiology, growth and resource management. A brief history of the utilization of this species, and an account of its chemical constituents including the hydrocolloid furcellaran are also presented.     

Down-modulation of epidermal growth factor receptor affinity in fibroblasts treated with interleukin 1 or tumor necrosis factor is associated with phosphorylation at a site other than threonine 654.

Interleukin 1 or tumor necrosis factor alpha can cause a transient down-modulation of epidermal growth factor (EGF) binding to quiescent fibroblast monolayers; the effect results from a reduction in EGF receptor (EGF-R) affinity and appears to be mediated by a protein kinase C (PKC)-independent mechanism. Here we show transient increases in EGF-R serine/threonine phosphorylation which are temporally coordinated with the effects on EGF binding; we also demonstrate that the cytokine-mediated phosphorylations, unlike those caused by PKC activators, have little discernible effect upon intrinsic EGF-R-associated tyrosine kinase activity. Cytokine-mediated EGF-R phosphorylation is resistant to staurosporine, an extremely potent inhibitor of PKC. Analysis of tryptic 32P-phosphopeptides reveals that Thr654, the unique site of PKC-mediated phosphorylation, is not phosphorylated in cytokine-treated cells, but a different, relatively acidic, peptide containing phosphoserine can be detected instead.     

Description of Meloidogyne nataliei n. sp. (Nematoda: Meloidogynidae) from Grape (Vitis labrusca) in Michigan,with SEM Observations

Meloidogyne nataliei n. sp. is described and illustrated from grape (Vitis labrusca) in a declining vineyard at Mattawan, Michigan, USA. Infected grape roots exhibit no hyperplastic symptoms. Females protrude from the roots and are surrounded by a massive egg sac containing many eggs. This new species is distinguishable from other species of the genus especially by its large, striking perineal pattern with, usually, two ropelike separated striae on each side extending laterally from the vulval and anal areas. Among other diagnostic characters are the location of the female excretory pore adjacent to or near the base of the head and the heavy larval stylet averaging about 22 μm in length. Examination of males, females, and larvae with the scanning electron microscope confirmed observations made by optical microscopy and revealed diagnostic and other structures in greater detail. Of particular significance was the nature of the male head, with a massive circular labial disc on which is located a rectangular structure surrounding the oral opening, and the six distinct lips appearing as a rosette in en face view. The known distribution of this new species is presently limited to its original location in Michigan.     

Plasma lipoprotein levels and the prevalence of hyperlipoproteinemia in a Canadian working population

The lipids and lipoproteins — cholesterol (C), triglyceride (TG) and high-density, low-density, very-low-density and sinking pre-β-lipoprotein cholesterol (HDL-C, LDL-C, VLDL-C and SPB-C) — in plasma samples from 1620 fasting white adults and children from the Toronto—Hamilton area were analysed. The mean concentration of HDL-C was about 45 mg/dl in men and about 60 mg/dl in women, and the levels were constant throughout adult life in both sexes. Boys had higher mean HDL-C levels than men, but girls had lower mean HDL-C levels than women. Mean LDL-C levels, like total C levels, increased with age, from about 87 mg/dl in boys to 136 mg/dl in men, and from about 91 mg/dl in girls to 145 mg/dl in women. The mean levels of VLDL-C followed the TG patterns for age and sex, rising from about 7 mg/dl in boys to 26 mg/dl in men, and from about 11 mg/dl in girls to 19 mg/dl in women. SPB-C was detectable visually in 39% of the population and with the aid of densitometry in 54%; the levels were not related to age, sex or oral contraceptive use, and the median level was 3 mg/dl.Prevalence estimates of hyperlipoproteinemia showed that type IV was the most common, and it was found more than three times as often in men as in women. This was in part due to the customary use of plasma TG cut-off points that do not reflect the large difference in TG levels between males and females. Type IIA hyperlipoproteinemia was found in about 2% of the adults and type IIb in a further 1%. Types I, III and V were all rare. The prevalence of types II and IV hyperlipoproteinemia was four times greater in women using oral contraceptives than in nonusers in the same age range.     

Localisation of thiamine pyrophosphatase within the cytoplasmic fine structure of trophozoites of Entamoeba histolytica and E. invadens

McCaul T.F. and Bird R.G. 1978. Localisation of thiamine pyrophosphatase within the cytoplasmic fine structure of trophozoites of Entamoeba histolytica and E. invadens. International Journal for Parasitology8: 501–506. The distribution of thiamine pyrophosphatase (TPPase) activity was studied in both formaldehyde and glutaraldehyde fixed trophozoites of Entamoeba histolytica and E. invadens. The activity was localised within certain vacuoles. No dense deposits for TPPase activity were seen in the small vesicles, elongated smooth-walled lacunae equated with endoplasmic reticulum, or the nucleus. The demonstration of small vesicles surrounding the larger vacuoles indicated that the Golgi-like vacuoles might be involved in the production of cell coat materials and primary lysosomes.