A new CSLM-based method for determination of the phase behaviour of aqueous mixtures of biopolymers

On mixing different types of high molecular weight (bio)polymers in an aqueous solution, phase separation often occurs. In some cases, the occurrence of phase separation may be readily observed, because due to density differences the heavier of the two phases is accumulated at the bottom of the vessel in which the mixture is contained. By using classical techniques, the composition of the two phases may then be determined. In the case where the density differences are not so large, and the viscosity of the system is high, the two phases remain intimately mixed. An alternative route to determine the phase behaviour of these systems might be a microscopic technique (Confocal Scanning Laser Microscopy, CSLM), using the fluorescence intensity of labelled biopolymers to quantify their concentration and phase volume in the system. Experiments were performed with several mixtures of sodium alginate, labelled with fluorescein, and sodium caseinate, fluorescently labelled with Texas Red. The viscosity of the mixtures studied was low enough to allow bulk phase separation of the phases by using an ultracentrifuge. Results of the phase volumes, and the composition of the phases, obtained independently by applying the two different methods (CSLM, or analysis of the separate phases after centrifugation) were compared and found to be in reasonable agreement.     

Changes in responsiveness to ethylene and gibberellin during corolla expansion ofIpomoea nil

Corolla expansion inIpomoea nil appears to be triggered by changes in gibberellin concentration and ethylene production during development. We investigated the role of responsiveness to GA and ethylene in corolla expansion. The effects of growth regulators applied in vitro were measured as a change in area of corolla segments from younger (15–17 mm) and older (18–20 mm) whole corollas. Applied gibberellic acid (GA₃) significantly (p < 0.05) promoted growth in the younger segments but was less effective in the older segments. Moreover, applications of the GA biosynthesis inhibitors, PP333 (paclobutrazol) AMO₁₆₁₈ (2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidinecarboxylate methyl chloride), chlorocholine chloride, and tetcyclasis had little effect on younger segments but inhibited growth of older segments. The older corollas have apparently synthesized and accumulated enough GA-like substances to become less responsive to additional applied GA₃. The amount of growth induced by applied or endogenous GA depended on the amount of ethylene simultaneously produced in the tissue. The younger corollas rapidly produced ethylene from endogenous 1-aminocyclopropane-1-carboxylic acid (ACC) and did not respond to applied ACC whereas the older corollas naturally produced much less ethylene and were significantly (p < 0.05) inhibited by applied ACC. When ethylene production was inhibited by applying aminoethoxyvinylglycine (AVG), growth was promoted in all segments. However, only the growth of the younger segments was further stimulated by simultaneously applied AVG and GA₃ over the GA₃ control. Thus the differential responses of segments from 15- to 20-mm long corollas to applied growth regulators reflect developmental changes in responsiveness of the developing corolla. The change in responsiveness is attributed in part to the changes in production of endogenous growth regulators and to the effect of one endogenous plant growth regulator (PGR) on the responsiveness of the corolla to another PGR.     

Non-radioactive detection of single-copy DNA-DNA hybrids

A new procedure for non-radioactive detection of single-copy DNA-DNA hybrids combines an existing non-radioactive labeling and detection kit with a new substrate AMPPD for the enzyme alkaline phosphatase. The main advantages of this procedure are the possibility to reuse the blots easily and the much shorter detection time compared to radioactive detection methods.     

Expression of C gamma 4 T cell receptors and lack of isotype exclusion by dendritic epidermal T cell lines

Although four murine C gamma gene segments (C gamma 1, 2, 3, and 4) are known to exist, the large majority of expressed gamma-chains have been shown to be of the C gamma 1 isotype and no evidence exists for the expression of more than one receptor by gamma delta TCR-bearing cells. We investigated the nature of the TCR expressed on a number of murine dendritic epidermal T cell-derived cell lines by using both Northern blot and immunoprecipitation analyses. One of these CD3+ cell lines (T195) expresses C gamma 4, V gamma 1, and delta mRNA, and its CD3-associated TCR complex can be precipitated by both anti-C gamma 4 and anti-delta sera, indicating that this receptor is a C gamma 4/delta heterodimer. Furthermore, we show that two cell lines (Y245, Y93) express two distinct TCR gamma-chains, one derived from the C gamma 4 locus, whereas the second gamma-chain is probably derived from the C gamma 2 locus. Together with the previous demonstration of C gamma 1/delta TCR on a number of dendritic epidermal T cell lines (DETC), these results indicate that such DETC are capable of expressing a variety of gamma delta TCR and that, in some DETC, isotype exclusion of gamma-chain expression does not occur.     

Characterization of a cell surface-expressed disulfide-linked dimer involved in murine T cell activation

We have produced a hamster mAb, H1.2F3, which was derived by immunization with a murine TCR-gamma delta + epidermal T cell line. H1.2F3 immunoprecipitates a cell surface-expressed disulfide-linked dimer that has a m.w. of 85,000 under non-reducing conditions and consists of subunits of 35,000 to 39,000 m.w. This dimer is distinct from the CD3-associated TCR-gamma delta complex (CD3/TCR), inasmuch as H1.2F3 does not co-precipitate or co-modulate with the CD3/TCR complex and recognizes an Ag with a single-peptide backbone of 22,000 m.w. after N-Glycanase treatment. H1.2F3 is weakly reactive with a small percentage of cells from unfractionated thymus, spleen, or lymph node, but reactivity with both T and B lymphocytes is markedly enhanced by a brief period of stimulation with Con A or PMA in vitro. This enhancement requires de novo protein synthesis. Enhanced expression of the H1.2F3 Ag can also be induced in vivo by injection of Con A or anti-CD3. H1.2F3 is a potent stimulator of T, but not B, cell proliferation in the presence of PMA and FcR-bearing accessory cells. These functional and biochemical studies strongly suggest that the Ag recognized by H1.2F3 is the murine homologue of the human CD28 Ag recognized by mAb 9.3.     

Prognostic factors in human primary breast cancer: comparison of c-myc and HER2/neu amplification.

Amplification of oncogenes in primary tumours may have prognostic and/or therapeutic significance for patients with breast cancer. We have studied HER2/neu and c-myc amplification together with steroid receptors in human primary breast tumours and related the outcome with (relapse-free) survival. A strong inverse correlation was found between HER2/neu amplification and the presence of oestrogen and progesterone receptors. Actuarial 5-years survival showed that breast cancer patients with c-myc amplification in their primary tumours experience a shorter relapse-free survival, especially in node-negative and in receptor-positive tumours, whereas HER2/neu amplification may be of prognostic value for overall survival in receptor-negative tumours. Overall, in our hands, c-myc amplification appeared to be a more potent prognosticator than HER2/neu amplification in human primary breast cancer.     

Four-hourly fluctuations in grass-pollen concentrations in relation to wet versus dry weather,and to short versus long over-land advection

Four-hourly volumetric measurements of airborne grass-pollen concentrations at Leiden, near the west coast of the Netherlands, were analyzed according to wet-versus-dry meteorological conditions in the pollen source area, and according to the distance of the over-land advection to the pollen sampler. Airborne pollen concentrations appear to be low when the source surface is wet by past or present rain, fog, or dew; and high when the source area is dry. Air adverted over long over-land distances from dry source areas contains much pollen, especially in the afternoon, due to pollen release and decreasing air turbulence. High nightly pollen concentrations are observed after a warm and dry day with much pollen release in distant source areas when the nocturnal meteorological conditions stimulate concentration of pollen grains into the lower layers of the atmosphere.Presented at the Sixth International Palynological Conference, 26 August – 1 September 1984, Calgary, Canada.     

L'action de l'air ionisé dans le traitement des ulcères gastroduodénaux (Données expérimentales chez le rat et essais cliniques)

Résumé Par un jeûne de trois jours ou par l'administration per os de réserpine(2 à 4 mg/kg) durant 15 à 24 jours, on a provoqué des ulcères d'estomac chez des rats. Une partie des bÊtes ainsi traitées fut exposée durant cette période et pendant 2 heures par jour à un air ionisé contenant 5.000 à 15.000 petits ions par cm³ d'air. Pour les animaux soumis au jeûne, la proportion d'ulcères se monte à 93% chez les bÊtes de contrÔle, alors qu'elle n'est que de 59% chez les bÊtes traÎtées par l'air ionisé. Pour les rats traÎtés à la réserpine, les proportions sont de 83,respectivement de 45%. On a en outre exposé 66 patients atteints d'ulcères d'estomac et d'intestins durant 12 à 15 jours et pendant 15 à 45 minutes par jour à un air contenant 5.000 à 10.000 petits ions négatifs par cm³ d'air. Pour 38 d'entre eux, la douleur et les autres symptÔmes avaient diminué ou disparu après le traitement.

---

Gastric ulcers were produced in rats by starvation for 3 days or the administration of reserpine (2 to 4 mg/kg) for 15 to 24 days. Some of the animals were exposed to ionized air for 2 hours daily during these periods(5,000 to 15,000 small negative ions/m3 air). The incidence of ulcer development was 93% in the untreated starved rats and 59% in those treated with ionized air; 83% in the animals treated with reserpine and 45% in those with additional ionized air treatment. In a clinical investigation 66 patients with peptic ulcer were exposed for 12 to 15 days to 5,000 to 10,000 small negative ions/m³ air daily. Pain and other symptoms were diminished or abolished after this treatment in 38 patients.

---

Zusammenfassung Bei Ratten wurden durch 3 Tage Hungern oder Behandlung mit Reserpin (2–4 mg/kg) wÄhrend 15 bis 24 Tagen Magenulcera provoziert. Ein Teil der Tiere wurde wÄhrend dieser Perioden 2 Std. tÄglich ionisierter Luft exponiert (5.000–15.000 kleine Ionen/m3 Luft). Die Ulcusrate betrug bei den unbehandelten Hungertieren 93%, bei den mit ionisierter Luft behandelten 59%; bei den Reserpin behandelten Tieren betrug sie 83% und bei den zusÄtzlich mit ionisierter Luft behandelten 45%. In der Klinik wurden 66 Patienten mit Magen-Darmulcera 12–15 Tage lang 15–45 min tÄglich mit 5.000 –10.000 neg. kleinen Ionen/m3 Luft behandelt. In 38 Patienten waren nach der Behandlung der Schmerz und die anderen Symptome abgeschwÄcht oder verschwunden.

     

City spore concentrations in the European Economic Community (EEC). VI. Poaceae (Grasses), 1982–1986

Summary Airborne grass-pollen concentrations in six cities in the EEC are compared, based on observations from five years, 1982–1986. Results show that there are quantitative differences both between the monitoring stations and between the years. Very provisionally, the average seasonal total of the European urban airborne grass-pollen concentration can be put at 4 to 5000 per m³ of air. Also qualitatively, regarding the seasonal fluctuations, there are differences between the stations, and between the years. Generally, the results confirm that June is the most typical grass-pollen month in northwestern Europe, whereas in mediterranean Europe May is the more prominent grass-pollen period. There seems to be little coherence between the starting dates of the grass-pollen season even at relatively nearby stations in northwestern Europe, suggestive for a great influence of the actual weather situation.