Evidence of Transcellular Strands in Transverse Cryostat Sections of Cucurbita pepo Sieve Tubes

Transverse cryostat sections of rapidly frozen vascular bundlesof Cucurbita pepo were viewed in a microscope with Nomarskioptics. Structural bodies were frequently observed in sievetube lumina and filling the sieve plate pores. The bodies consistof an outer boundary ring and an inner core, often granularin appearance, and may represent transverse sections of boundedtubes filled by substructural material. This evidence is consistentwith earlier observations of strand-like structures with parallelsubstructural elements seen in longitudinal sections.     

MICROTUBULES IN THE SHELL OF THE INVASIVE BIVALVE CORBICULA FLUMINEA (MULLER, 1774) (BIVALVIA: HETERODONTA)

Using scanning electron and histological techniques on specimensof the bivalve Corbicula fluminea a new relationship betweenmantle, shell and periostracum has been observed, apparentlyfor the first time. Here we demonstrate that several extensionsof the mantle epithelium pierce the shell to join the innerlayer of the two-layered periostracum. The mantle extensionsare confirmed as unicellular processes. We suggest that theycould serve the animal in the mobilization of calcium from theshell for buffering the extrapalhal fluid under anaerobic conditions,when organic acids accumulate or when an extra contributionof Ca²⁺ is required (Received 5 January 1994; accepted 30 March 1994)     

Comparison of TRPs From Murine and Human Malignant Melanocytes

Most of our knowledge of the mammalian tyrosinase related protein (TRP) activities is derived from studies using murine melanoma models, such as B16 or Cloudman S-91 melanocytes. Owing to the high degree of homology between the murine and human enzymes, it has been assumed that their kinetic behavior could be similar. However, the protein sequences at the metal binding sites of the murine and human enzymes show some differences of possible functional relevance. These differences are more significant in the metal-A site than in the metal-B site. By using three human melanoma cell lines (HBL, SCL, and BEU), we have studied the catalytic abilities of the human melanogenic enzymes in comparison to those obtained for the counterpart murine enzymes isolated from B16 melanoma. We have found that TRP2 extracted from all cell lines show dopachrome tautomerase activity, although the activity levels in human malignant melanocytes are much lower than in mouse cells. Reconstitution experiments of the human enzyme indicate that TRP2 has Zn at its metal binding-sites. Although mouse tyrosinase does not show DHICA oxidase activity, and this step of the melanogenesis pathway is specifically catalyzed by mouse TRP1, the human enzyme seems to recognize carboxylated indoles. Thus, human tyrosinase could display some residual DHICA oxidase activity, and the function of human TRP1 could differ from that of the murine protein. Attempts to clarify the nature of the metal cofactor in TRP1 were unsuccessful. The enzyme contains mostly Fe and Cu, but the reconstitution of the enzymatic activity from the apoprotein with these ions was not possible.     

Estrogen and Androgen Target Cells in the Brain of Fishes, Reptiles and Birds: Phylogeny and Ontogeny

The cellular and subcellular distribution of 3H estradiol inthe forebrain of goldfish, platyfish, lizard, turtle, and ringdove was studied by thaw-mount autoradiography. The topographicdistribution of estrogen target cells in these non-mammalianspecies corresponds to the general pattern established earlierin our laboratory for mammals. In all non-mammalians studied,accumulations of estrogen target cells exist in preoptic-septal,central hypothalamic and thalamic areas. While allocorticaldistribution of estrogen target cells is present in the avianand reptilian species, it appears to be absent in the teleosts.Differences in brain evolution probably account for it. Thisphylogenetically delayed acquisition of telencephalic steroidtarget sites seems paralleled by an ontogenetic delay, as observedin the chick embryo: early embryonic appearance of estrogenuptake (receptors) in preoptic and central hypothalamic regionsand late appearance of estrogen receptors in the amygdala. Androgendistribution in the forebrain of the dove and lizard overlapswith the estrogen distribution with some exceptions.     

Amplification of a New Region of DNA in an Unselected Laboratory Stock of L. tarentolae: The T Region

A new DNA amplification is described from an isolate of the lizard parasite Leishmania tarentolae . This DNA is present in up to 50 copies in the Trager line of this species and present but not amplified in all other lines tested. This amplification has been named the T amplification (for Tarentolae/Trager). Restriction enzyme digestion and electrophoresis of total DNA reveal amplified fragments totalling 19 kb following staining with ethidium bromide, a finding confirmed by the use of specific hybridization probes. Much of the amplified T DNA occurs as extra-chromosomal circular molecules. No cross-hybridization was observed between the T region and other amplified DNA of Leishmania , or the maxicircle of L. tarentolae , nor was resistance to methotrexate, chloroquine or primaquine detected in the T-amplified line. Combined with our previous results showing H region amplification in 2 other unselected lab stocks, these data demonstrate the prevalence of apparently spontaneous gene amplifications in L. tarentolae .     

New stingless bee genus endemic to Central American cloudforests: phylogenetic and biogeographic implications (Hymenoptera: Apidae: Meliponini)

Meliwillea bivea gen.n., sp.n. is the only known stingless bee genus endemic to Central America. External morphology and male genitalia show affinity of Meliwillea with Scaptotrigona . Its plesiomorphies and current sympatry with Scaptotrigona suggest Meliwillea is relictual and diverged in montane habitat during the Tertiary, predating the Pleistocene connection between Central and South America which allowed immigration by Scaptotrigona . Addition of Meliwillea to the list of Neotropical genera changes the phylogenetic position of Nannotrigona , making it sister to Paratrigona instead of Scaptotrigona .     

Lorica Construction in Eufolliculina sp. (Ciliophora,Heterotrichida)1

ABSTRACT. The structure and ultrastructure of the chitinous lorica of Eufolliculina sp. are described. The lorica is produced from precursor material secreted by the motile swarmer immediately after settling. This material is located in numerous vesicles found in the cortical region of the cells and is secreted by exocytosis. Initially, material is secreted from the ventral part of the cell to produce the attachment plate of the lorica. After this, exocytosis occurs over most of the body surface as the ampulla part of the lorica is constructed. During the later stages of lorica formation, secretion is mainly limited to the anterior of the cell as the neck is formed. The lorica is shaped mainly by the action of the cilia and by the behavior of the cell. While the neck is being formed, the anterior part of the cell is deformed by a local accumulation of cytoplasmic vacuoles. This deformation is employed in shaping the neck. No changes were detected in the organization of the cortical infraciliature during the first stages of lorica formation, but they do occur after the neck has been produced and as the swarmer develops into the sessile form.     

Comparison of Coding and Spacer Region Sequences of Chromosomal rRNA-Coding Genes of Two Sequevars of Pneumocystis carinii

Two distinct sequevars, denoted Pc1 and Pc2, of the opportunistic pathogen Pneumocystis carinii have been previously identified based on the sequence of their 26S rRNA genes, the location of group I self-splicing introns and pulsed field electrophoretic patterns of chromosomal DNA. This study shows that the sequences of 16S and 5.8S rRNA genes also vary between these sequevars, and that greater variation was seen in the internal transcribed spacer regions. Polymerase chain reaction and restriction analysis can distinguish between these sequevars.