α-Globin gene deletion causes α-thalassemia syndromes in two German families

Summary Restriction endonuclease mapping of chromosomal DNA has been used to determine whether the -globin gene deletion or non-deletion form of -thalassemia is the underlying molecular defect in individuals of two unrelated German families with -thalassemia syndromes. The obtained DNA pattern in all cases indicated loss of -globin genes resulting in-/,--/, and--/- genotypes in thalassemia-2, -thalassemia-1, and Hb H individuals respectively. The chromosomes showing loss of one -globin gene in -thalassemia-2 and Hb H disease were characterized by the so-called rightward deletion form exhibiting loss of a 3.7 kb DNA fragment in the -gene cluster.     

The composition of Erythrosins, Fluorescein, Phloxine and Rose Bengal: a study using thin-layer chromatography and solvent extraction

Synopsis Commercial samples of Erythrosin B (CI 45430), Erythrosin Y (CI 45425), Fluorescein (CI 45350), Phloxine (CI 45410) and Rose Bengal (CI 45440) have been analysed by thin-layer chromatography. The Erythrosins were found to be mixtures consisting in the main of 4-iodofluorescein, 4,5-di-iodofluorescein, 2,4,5-triiodofluorescein and 2,4,5,7-tetraiodofluorescein, in some instances together with 2,4,5-tri-iodo-4,5,6,7-tetrachlorofluorescein and 2,4,5,7-tetraiodo-4,5,6,7-tetrachlorofluorescein. Samples of Fluorescein were mixtures of the nominal dye usually with traces of several unidentified, fluorescent components. Those of Phloxine consisted mainly of mixtures of 4-bromo-4,5,6,7-tetrachlorofluorescein, 4,5-dibromo-4,5,6,7-tetrachlorofluorescein, 2,4,5-tribromo-4,5,6,7-tetrachlorofluorescein and 2,4,5,7-tetrabromo-4,5,6,7-tetrachlorofluorescein, often with 4,5,6,7-tetrachlorofluorescein Samples of Rose Bengal were mixtures of 4-iodo-4,5,6,7-tetrachlorofluorescein, 4,5-di-iodo-4,5,6,7-tetrachlorofluorescein, 2,4,5-tri-iodo-4,5,6,7-tetrachlorofluorescein and 2,4,5,7-tetraiodo-4,5,6,7-tetrachlorofluorescein together with some unidentified components.Most of the commercial dye samples gave an insoluble residue when extracted with methanol. This residue was usually inorganic carbonate or halide. Some possible practical consequences of the various impurities are discussed.     

Dephosphorylated oligoadenylates modulate high voltage-activated calcium currents in GH3 cells

Effects of different forms of C2-5A (2,5ApApA; 2,5ApApepoxyA; and 3,5ApApA) on high voltage-activated (HVA) calcium currents in GH3 cells were studied using the whole-cell patch-clamp recording technique. Addition of 10 µM 2,5ApApA, a core (dephosphorylated) oligoadenylate, to the pipette solution induced an increase in HVA calcium current. Ten minutes after the whole-cell configuration was established, the current magnitude was enhanced about twofold compared with that observed at 2 min. High concentration of Mg²⁺ (5 µM) in the pipette solution blocked this effect. 2,5ApA and 3,5ApApA oligoadenylates, and products of 2,5A hydrolysis, adenosine and AMP, did not change the value of HVA current. A chemically modified analog of 2,5ApApA (2,5ApApepoxyA) has been the oligoadenylate most stable under phosphodiesterase action. Addition of 2,5ApApepoxyA to the pipette solution under the same conditions caused a smaller effect than 2,5ApApA did.Neirofiziologiya/Neurophysiology, Vol. 26, No. 6, pp. 405–408, November–December, 1994.     

Diurnal Lhc gene expression is present in many but not all species of the plant kingdom

The diurnal and circadian expression of light-harvesting genes (Lhc) is well documented for many plant species of the Angiospermae division. Here we present the diurnal mRNA levels of species of the Gymnospermae, Pteridophyta, Bryophyta and Phycophyta divisions. Except for four Coniferophytina species, diurnal Lhc mRNA accumulation is detected in fern, moss and algae, supporting the idea that the concept of circadian clock-controlled gene expression is an ancient process. Possible reasons why plants need the circadian clock control mechanism are discussed.Dedicated to Dr H. W. Heldt on the occasion of his 60th birthday.     

Sequence analysis of linked maize 22 kDa α-zein genes

We have determined the nucleotide sequence of a 7343 bp zein genomic clone (gZ22.8H3) from the maize inbred W64A. Computer-aided analysis of the DNA sequence revealed two contiguous 22 kDa -zein genes. The 5 gene (gZ22.8) encodes a complete polypeptide and contains putative regulatory sequences in both the 5 and 3 flanking regions that are typical of zein genes. In contrast, the 3 gene (gZ22.8) appears to be a pseudogene, because it contains numerous insertions and deletions that would prevent translation of the mRNA. Alignment of the 5 and 3 flanking sequences of both genes indicated that they resulted from a 3.3 kb DNA duplication event.     

Conformational transitions in closed circular DNA molecules I. Topological and energetical considerations

A theory of conformational transitions in closed circular DNA as a function of topological linking number of the molecule () is elaborated taking into account topological and energetical considerations. The theory predicts a step-like dependence of a number of superhelical turns in DNA molecules () on . Thus, the number of superhelical turns = for small values of . For a large (when conformational transitions begin to occur) =–_ij, where _ij is the total angle of conformational transitions for a given . This prediction is in good agreement with published data on the dependence of the sedimentation coefficient of circular DNA molecules on their topological linking number. The results also allow to explain the disagreement between a number of titratable superhelical turns in circular DNA molecules and a number of supercoiles seen on electron micrographs for molecules with sufficiently large .     

The nutritional status of Picea abies (L.) Karst. across Europe,and implications for ‘forest decline’

Summary During the summer of 1986, three year-classes of foliage were sampled from approximately 30-year-old Norway spruce [Picea abies (L.) Karst.] trees at 12 sites from S. W. Germany to N. Scotland. At sites in Germany, where trees were showing symptoms of decline, samples were taken from trees with good crown condition and poor crown conditon. The distinction between good and poor was made on the basis of international protocols for defining crown density and foliar discoloration. There was a wide range in nutrient content (percent dry weight) in apparently healthy trees. Current year foliage had ranges of mean values per site: S(0.07–0.13%), N(0.9–1.4%), K(0.5–0.9%), Ca(0.2–0.7%), and Mg (0.05–0.1%). Ranges were greater for 2-year-old foliage: S(0.09–0.18%), N(1.0–1.8%), K(0.4–0.7%), Ca(0.2–1.4%), and Mg(0.03–0.09%). At sites with trees having poor crown condition, there were significantly smaller concentrations of Mg and Ca, and larger concentrations of K in 2-year-old foliage from poor trees, compared with adjacent good trees. Ratios of nutrient content were more significantly related to crown condition within sites than individual nutrients, especially in older needles. Poor crowns were associated with larger ratios of NMg, KMg, SMg, KCa and smaller ratios of SK and NK. A risk index is defined for trees showing no visible decline symptoms, based upon nutrient content and nutrient ratios, which may be useful in identifying sites liable to experience deterioration in crown condition.With the exception of one German site, where few poor trees were observed, the index increases from Scottish sites to English sites to Dutch sites to German sites. The index is empirical, and not necessarily related to potential effects of air pollution. The time dependence of foliar nutrient content may also be useful in diagnosis. At sites with trees having poor crown condition, even apparently healthy trees showed a lack of increase in calcium content with needle age, decreases in nitrogen content and very large decreases in magnesium content with needle age. The results show the importance of sampling several year classes of foliage from Norway spruce trees in determining the nutrient status of the tree.     

The effects of dissolved macro-nutrients on the herbaceous vegetation around dune pools

In Dutch coastal dunes the relation was studied between the composition of the vegetation on the banks of pools and the nutrients supplied by the groundwater. Areas infiltrated with eutrophic river water were compared with non-affected areas. The floristic composition was interpreted according to classification systems of ruderal, nitrophilous tall hemicryptophytes and the conservation parameters richness and rarity. Of the three main macro-nutrients (orthophosphate, nitrate and potassium), the orthophosphate load shows the highest correlation with the vegetation variables. It is suggested that a reduction of orthophosphate during pre-purification of the infiltration water would reduce the dominance of the ruderals to the extent that the original dune slack species could return locally.Nomenclature of species follows Arnolds & Van der Meijden (1975).We thank G. J. de Bruyn, W. H. van Dobben, W. T. de Groot, W. J. ter Keurs, F. van der Meulen, W. H. van der Molen, M. A. W. Noordervliet, and I. S. Zonneveld for their comments on the study; J. Braak, J. Brakkee and R. van Nood for their assistance with the chemical analyses; A. R. Kaal for the translation, and G. P. G. Hock for the drawing of the figures.     

Spectral position control of dye absorption band maximum in an orienting matrix

The continuous control of the maximum position of the dye absorption band (the zero of the derivative dD ()/d of the cell's optical density D ()) in a nematic matrix is demonstrated experimentally, as a result of changing the angle between the optical axis of a planar-oriented sample and the plane of polarization of absorbed light incident normal to the optical axis. The theory proposed describes quantitatively the experimental dependence (). The rotation of the polarizer with given frequency results in the spectral position modulation of the solute band maximum () within (=0°)–(90°)=700 cm^–1.     

Green manures of oat,rape and sweet corn for reducing common root rot in pea (Pisum sativum) caused by Aphanomyces euteiches

Green manure crops of sweet corn, soybean, alfalfa, snap bean, rape, pea and of the two oat cultivars Dane and Troy were incorporated into the same soil and containers in which the crop had grown for five weeks. The soil was then evaluated for suppression of common root rot (Aphanomyces euteiches) of pea grown in infested pasteurized and non-pasteurized soils in the greenhouse. Pea biomass reduction and a plant bioassay for A. euteiches were used to measure the green manure suppression of disease. Green manures of sweet corn cv.Jubilee, oat cv.Troy, and rape cv.Humus significantly reduced pea biomass losses over the non-amended control soil treatments. Oat cv.Troyand sweet corn cv.Jubilee green manures significantly reduced inoculum density of A. euteiches over the corresponding fallow controls in inoculated pasteurized soil by 87% and 76%, respectively, and in inoculated non-pasteurized soil by 67% and 66%, respectively. Only the green manure of oat cv. Troy reduced inoculum density significantly below fallow.     

Improved rate of callus induction from rice anther culture following microscopic staging of microspores in iron alum-haematoxylin

Summary High frequencies of callusing were obtained in indica rice from the microspores which were staged in acetic acid iron alum-haematoxylin stain prior to culture on G5 medium. Two local varieties, Khonorullo and Namyi, and two advance pre-release cultures, PK 1-1-3 and PK 12-22, were used in this investigation. All the cultures exhibited a wide adaptation to varying medium; however, the frequency of callusing was highest (45.5%) in PK 1-1-3 followed by PK 12-22 (32.4%) and Khonorullo (31.6%). Cold shock (10 °C) for 11 days enhanced the frequency of callusing by 200% in Khonorullo.     

An alfalfa (Medicago sativa L.) cDNA encoding an acidic leghemoglobin (MsLb3)

We have found an alfalfa cDNA clone that encodes an acidic leghemoglobin. To date, 14 alfalfa leghemoglobin clones have been identified. Five different leghemoglobin components have been biochemically defined on the basis of their pI. A higher-resolution comparison, provided by sequence data analysis, identifies six leghemoglobin classes. All 14 leghemoglobins are assigned to the six classes, which can be distributed among the five leghemoglobin components. The newly identified leghemoglobin is the only member of a sixth class of leghemoglobins, and it also is the only member of one of the acidic leghemoglobin components IV or V.     

Involvement of abscisic acid in bulb dormancy of Allium wakegi Araki. II. A comparison between dormant and nondormant cultivars

The content of abscisic acid (ABA) in bulbs of two Allium wakegi Araki cultivars, Kiharabansei No. 1 (dormant type) and Ginoza (nondormant type), was similar and changed similarly during the development and storage of the bulbs. It increased during bulb development, reached a maximum shortly after bulb harvesting, and gradually decreased during bulb storage. The bulbs of Kiharabansei No. 1 showed dormancy correlated with the change in ABA content, but those of Ginoza did not show significant dormancy throughout the experimental period. The ABA content in the buds of dormant bulbs of Kiharabansei No. 1 did not change after planting of bulbs, but that of nondormant bulbs of Ginoza planted on the same day rapidly decreased after planting. Application of ABA to bulbs delayed sprouting of both cultivars, but dormant bulbs of Kiharabansei No. 1 had higher sensitivity to ABA than the bulbs of Ginoza or the bulbs of Kiharabansei No. 1 partly released from dormancy. These results suggest that the decrease in the ABA content after planting (watering) and low sensitivity to ABA are correlated with the nondormancy of Ginoza.     

A study on relations of vegetation,climate and soils in Shanxi province,China

Relationships of vegetation, climate and soils in Shanxi plateau wereanalyzed by use of Canonical Correspondence Analysis (CCA). Shanxi province,located at 34°35–40°43 N, 110°15–114°33 E, was divided into a series of rectangular districts of30 latitude by 20 longitude. Areas of vegetation formations and soil types ineach district were measured carefully using fine grain on the vegetation andsoil maps of Shanxi. Climatic data were mean values of 25 years records in eachdistrict. Three data matrices of climate, vegetation and soil were combined byCCA. The results showed that the distribution of vegetation is closely relatedto the variety of climates and to soils distribution.     

Modulation of cytokine responses of murine CD8+ αβ intestinal intraepithelial lymphocytes by IL-4 and IL-12

The immune responses of the intestine mucosa feature the noninflammatory type, such as IgA production and oral tolerance. Th2 type cytokines have been implicated in the induction of these noninflammatory responses. In the present study, cytokine responses of CD8+ and CD4+ TCR+ intestinal intraepithelial lymphocyte ( iIEL) subsets to TCR stimulation under the influence of IL-12, IL-4, or CD28 costimulation were examined. IL-12 enhanced production of IL-10 and IFN- by the CD8+ iIEL significantly but only marginally affected the CD8+ subset, whereas IL-4 induced IL-4, IL-5, and IL-10 production and augmented TGF- production by both subsets. CD28 costimulation induced production of Th2 cytokines by CD4+ iIEL in the absence of exogenous IL-4. Unlike lymph node CD4+ cells, the CD28 costimulation-induced Th2 differentiation of CD4+ iIEL was not inhibited by IFN-. These results demonstrate active cytokine production by CD4+, CD8+, as well as CD8+ iIEL. The Th2-skewed cytokine profile of CD8+ iIEL and the IFN--resistance of Th2 differentiation of the CD4+ iIEL suggest that both iIEL subsets contribute to the induction of noninflammatory mucosal immune responses.     

Promotion of murine antitumour activity by prothymosin α treatment: I. Induction of tumoricidal peritoneal cells producing high levels of tumour necrosis factor α

Summary The effect of prothymosin (ProT) on the survival of DBA/2 mice inoculated with syngeneic tumour cells was studied. DBA/2 mice inoculated intraperitoneally (i.p.) with 2×10⁵ syngeneic leukaemic L1210 cells developed ascites within 8–12 days and died 10–14 days later. Treatment with ProT consistently inhibited the development of ascites in 20% of the treated animals and prolonged the survival of 40%–60% of the animals up to 70 days. The most effective treatment schedule of ProT was 300 ng/mouse given i.p. at 2-day intervals for 3 weeks followed by a rest period of 7 days, prior to tumour cell inoculation. Peritoneal exudate (PE) cells collected from mice treated with the optimal dose of ProT produced, in the absence of exogenous stimulus, six- to eightfold higher levels of tumour necrosis factor (TNF) than PE cells from control mice. Furthermore these cells exhibited cytotoxic activity against several tumour cell lines including the syngeneic L1210, the TNF-insensitive P815 mastocytoma, the human MOLT-4 lymphoblastic leukaemia, as well as the murine TNF-sensitive L929 fibroblast cell line. Kinetic studies revealed that both production of TNF and tumoricidal activity peaked 7 days after the last injection of ProT and were maintained at high levels over a period of 1 month. Injections with 150 ng ProT slightly improved the survival of mice whereas higher (500 ng and 1000 ng) doses of ProT and a wide range of thymosin 1 doses remained without any effect. PE cells collected from these mice produced extremely low levels of TNF and exhibited negligible tumoricidal activity. Our data demonstrate that ProT has a protective effect in vivo against the growth of adoptively transfered tumour cells and suggest that this effect is, at least in part, mediated by ProT-activated PE cells. These cells were demonstrated to produce high levels of TNF in vitro and to exhibit activity against both TNF-sensitive and TNF-resistant cell lines.Supported by a CEC grant to Dr. M. Papamichail     

The reduction of tetrazolium salts by plant mitochondria

Summary Data has been obtained concerning the reduction of tetrazolium salts by mitochondria isolated from Jerusalem artichoke tubers with succinate as the substrate using a direct recording spectrophotometric method of assay. ATP was found to increase the rate of reduction of the tetrazolium salts, this being independent of the effect ATP had on the rate of oxygen uptake. The magnitude of the stimulation by ATP depended on the concentration of tetrazolium salts present and under certain circumstances was suppressed by the addition of azide and cyanide. The sites at which the tetrazolium salts were reduced along the electron transport chain were investigated. The role of ATP has been discussed in relation to the mechanism of tetrazolium reduction.Abbreviations TTC 2,3,5-triphenyl-2,1,3,4-tetrazolium chloride - BT 5,5-diphenyl-3,3-(3,3-dimethoxy-4,4-diphenylene)-ditetrazolium chloride - NT 2,2,5,5-tetraphenyl-3,3-(p-diphenylene)-ditetrazolium chloride - MTT 3-(4,5-dimethyl thiozolyl-2)-2,5-diphenyl tetrazolium bromide - INT 2-(p-iodophenyl)-3-p-dinitrophenyl-3-p-nitrophenyl-5-phenyl tetrazolium chloride - NBT 2,2-dinitrophenyl-5,5-diphenyl-3,3-dimethoxy-4,4-diphenylene)-ditetrazolium chloride - TNBT 2,2-5,5-tetra-p-nitrophenyl-3,3-dimethoxy-4,4-diphenylene) ditetrazolium chloride     

Effect of photoperiod on in vitro tuberization of potato (Solanum tuberosum L.)

Single-node cuttings of potato cultivars Jemseg, Katahdin, Russet Burbank and Superior were cultured on a multiplication medium containing MS salts and no growth regulators. Cultures were exposed to 8 h (SD) and 16 h (LD) photoperiodic regimes. The subsequent plantlets were excised and single node cuttings from each photoperiodic regime were placed under SD or LD on a second medium containing growth regulators which promoted tuberization. Production of microtubers was strongly influenced by genotype and by photoperiodic treatments. Superior produced stunted plantlets and some microtubers under SD conditions in the multiplication medium. The number of microtubers formed by Jemseg was not influenced by photoperiod. However, Katahdin and Russet Burbank formed fewer microtubers under LD-LD conditions compared to LD-SD, SD-SD and SD-LD regimes. Compared with the other regimes, LD-SD photoperiod generally promoted microtuber formation with larger diameters and significantly (p<0.05) greater fresh weight. The intensity of the tuberization stimulus was affected by daylength, and this was characterized by microtubers with secondary tubers, the growth of more than one axillary microtuber, and microtubers subtended by stolons. The maturity group of the potato cultivars and photoperiodic regime in vitro strongly influenced the production of microtubers. These results can be employed to adapt light regimes for multiplication and tuberization to the specific requirements for cultivars from different maturity groups, and thus increase the efficiency of potato multiplication protocols.     

Effect of nitrogen on thiocyanate content of Brassica oleracea var. acephala leaves

An experiment was carried out to study the effect of nitrogen fertilizer application levels (0, 47, 94 and 188 kg N/ha) on thiocyanate contents of petioles and lamina of two Brassica oleracea L. var. acephala varieties (Thousand-headed kale and Georgia collards).The results showed that N application significantly decreased thiocyanate contents of both laminae and petioles. Applying 47, 94, 188 kg N/ha reduced thiocyanate of laminae and petioles by 26, 41 and 52% and 2, 9 and 39%, respectively. The results also showed that thiocyanate of leaves from Thousand-headed kale was significantly higher than that of Georgia collards. Lamina and petiole thiocyanates of Georgia collards were 79% and 83% of those of Thousand-headed kale, respectively.