Local cytoplasmic calcium gradients in living mitotic cells

Cytoplasmic free calcium has been proposed as a regulator of many microtubule-mediated processes, including mitosis. It has been difficult to test this hypothesis because methods for local measurement of free Ca2+ in the living cell have not been available. We have used the fluorescent calcium indicator dye Quin-2 (methoxyquinoline-1bis(o-aminophenoxy)ethane-N,N,N',N' -tetra acetic acid), which allows such observations to be made by digital processing of fluorescent images from the light microscope. Here we report the application of this technique to the study of local Ca2+ concentrations in mitotic endosperm cells of Haemanthus sp., and show that there is transient increase in free Ca2+ at the mitotic spindle poles during anaphase. This locally high Ca2+ may provide a mechanism for the regional control of microtubules and other cytoskeletal elements during anaphase.     

Calcium uptake by sarcoplasmic reticulum from nerve-intact and standard skeletal muscle grafts

A freely grafted rat soleus muscle exhibits a decrease in velocity and capacity of SR calcium uptake. This deficit is not prevented by maintaining neural connections (nerve-intact graft) during grafting. Thus the greater mechanical capability of nerve-intact grafts, relative to standard grafts, is not accompanied by any enhancement of the SR tubules.     

Human gamma-chain genes are rearranged in leukaemic T cells and map to the short arm of chromosome 7

Three gene families that rearrange during the somatic development of T cells have been identified in the murine genome. Two of these gene families (alpha and beta) encode subunits of the antigen-specific T-cell receptor and are also present in the human genome. The third gene family, designated here as the gamma-chain gene family, is rearranged in murine cytolytic T cells but not in most helper T cells. Here we present evidence that the human genome also contains gamma-chain genes that undergo somatic rearrangement in leukaemia-derived T cells. Murine gamma-chain genes appear to be encoded in gene segments that are analogous to the immunoglobulin gene variable, constant and joining segments. There are two closely related constant-region gene segments in the human genome. One of the constant-region genes is deleted in all three T-cell leukaemias that we have studied. The two constant-region gamma-chain genes reside on the short arm of chromosome 7 (7p15); this region is involved in chromosomal rearrangements identified in T cells from individuals with the immunodeficiency syndrome ataxia telangiectasia and observed only rarely in routine cytogenetic analyses of normal individuals. This region is also a secondary site of beta-chain gene hybridization.     

Androstenedione or corticosterone treatment during pregnancy alters estrous cycle of adult female offspring in mice

Female offspring from mice injected with androstenedione during late pregnancy showed lengthened vaginal cycles, persistent estrus and decreased incidence of pro-estrus and diestrus, whilst offspring from mice injected with corticosterone showed increased incidence of diestrus. These observations give qualified support to the hypothesis that stress during pregnancy alters the female offspring reproductive system through the action of adrenal steroids.     

Primary structure and transmembrane orientation of the murine anion exchange protein

The amino-acid sequence of murine band 3, deduced from the nucleotide sequence of a complementary DNA clone, confirms that this integral membrane glycoprotein is composed of two major structural domains which correlate with its dual functions as the anchor for the erythrocyte cytoskeleton and as a plasma membrane anion antiporter. This latter activity resides within a highly hydrophobic domain that crosses the plasma membrane at least 12 times.     

An HTLV-III peptide produced by recombinant DNA is immunoreactive with sera from patients with AIDS

Human T-cell lymphotropic retrovirus type III (HTLV-III), also called lymphadenopathy-associated virus (LAV), has been identified as the aetiological agent of acquired immune deficiency syndrome (AIDS). The sera of most patients with AIDS or AIDS-related complexes, and of asymptomatic individuals infected with HTLV-III, contain antibodies against antigens of HTLV-III. The characterization of these antibodies and their corresponding viral antigens is important not only for understanding immunity against HTLV-III and the pathology of AIDS, but also for the development of diagnostic methods and preventive vaccine for AIDS. Following the successful establishment of a long-term T-cell line permissive for HTLV-III replication, large quantities of virus have been produced, facilitating the purification of viral proteins and the development of mouse monoclonal antibodies against several viral antigens. More recently, the structure of HTLV-III proviral DNA has been elucidated. We now report the production, by genetic engineering methods, of a peptide encoded by a gene segment of HTLV-III. A 1.1-kilobase (kb) EcoRI DNA segment from an isolate of HTLV-III was inserted into a lpp and lac promoter-coupled expression vector, pIN-III-ompA. Escherichia coli transformants of this plasmid produced a peptide of relative molecular mass (Mr) 15,000 (15K) which was strongly immunoreactive with anti-HTLV-III antibodies present in sera from AIDS patients. Lysates of the clones expressing this 15K peptide inhibited the reactivity of the p31 virion protein with AIDS sera, suggesting that it is a fragment of the viral p31 protein. The peptide reacted with sera from all 20 AIDS patients but none of the 8 normal controls tested. These results suggest that the peptide may be useful for detecting anti-HTLV-III antibodies in blood samples.     

Molecular cloning of the fibroin light chain complementary DNA and its use in the study of the expression of the light chain gene in the posterior silk gland of Bombyx mori

Fibroin light chain (L-chain) mRNA (mol. wt 4.0 X 10(5) daltons) was purified from the posterior silk gland of the silkworm, Bombyx mori (J-131 strain). Double-stranded complementary DNA was synthesized and inserted into the PstI site of pBR322 employing the oligo(dC)-oligo(dG) tailing method. Several recombinant plasmids containing the inserts of about 800 base pairs were isolated. Hybridization-translation assay demonstrated that these clones hybridized specifically with the fibroin L-chain mRNA. One of these clones (pLA23) was used as a probe to investigate relative concentrations of the fibroin L-chain gene and mRNA in the posterior silk glands at different stages of late larval development.     

Seasonality in the reproductive phenology ofFicus: Its evolution and consequences

Summary Ficus can only reproduce if they are pollinated by mutualistic wasps that breed within the figs. Pollen-loaded wasps enter the figs when the female flowers are receptive. Several weeks later, their offspring load pollen within the fig and then emerge. As individual trees typically produce crops of synchronous figs at long intervals, the shortlived wasps have to move to another, receptive, tree. The wasp population can only survive, and hence the fig population reproduce, if there are trees fruiting all over the year. When only few trees are present within a population gaps in the flowering sequence may lead to the extinction of the local pollinator population. Two models are presented. One investigates the number of trees necessary in order to sustain a local pollinator population when the tree population has a seasonal pattern of fruiting. The second model investigates how such a seasonal pattern may evolve within a fig population as a result of individual selection on the trees. It is shown that pollinator populations are fragilized under seasonal conditions. Hence, the breeding system ofFicus limits their expansion into highly seasonal habitats. Seasonal habitats may also lead to seasonal adjustment of male versus female investments and to the evolution of dioecy.     

Streptozotocin induced diabetic nephropathy and renal tumors in the rat

Summary Adult Wistar rats rendered diabetic by a single dose of streptozotocin develop renal morphological changes which show subtle differences compared to those seen in human diabetic renal disease. The early tubular degeneration is sited in the distal rather than the proximal convoluted tubule and subsequent glomerular lesion shows linear deposits of IgG and albumin in the basement membrane rather than in the mesangium. The carcinogenicity of streptozotocin in the rat is reconfirmed.Acknowledgements. We wish to thank Mr S.G. Watkins and Mrs D. Greening for technical assistance.     

Effect of high anodic polarization on the passive layer properties of superduplex stainless steel friction stir welds at different chloride electrolyte pH values and temperatures

The conditions used for friction stir welding of duplex stainless steels determine the resulting mechanical and corrosion performance of the material.This study investigates the corrosion resistance of UNS S32750 and S32760 superduplex stainless steels(SDSSs)joined by friction stir welding,employing cyclic polarization,Mott–Schottky,and microscopy techniques for analysis.The microscopy images indicated the presence of a deleterious intermetallic phase after electrolytic etching of S32760,as well as decreased corrosion resistance.The presence of molybdenum in the steels promoted better passive behavior at low pH.The Mott–Schottky curves revealed p-n heterojunction behavior of the passive oxide.Images acquired after the polarization test by scanning electron microscopy showed higher passivation propensity with increases of temperature and pH.