Larval stages of Neodiaptomus lindbergi Brehm, 1953 (Copepoda,Calanoida)

All the postembryonic developmental instars of Neodiaptomus lindbergi Brehm, 1953, reared in the laboratory from eggs, are described and illustrated. Further, the larval affinities of this species vis-a-vis certain other Indian diaptomid species are mentioned.     

Synthesis of glycuronamides of amino acids,constituents of microbial polysaccharides and their conversion into neoglycoconjugates of copolymer type

Glycopyranosiduronic acids, amidically linked to amino acids (alanine, serine, threonine, and lysine) were prepared.O-tert-Butyl andN-tert-butyloxycarbonyl protected amino acidtert-butyl esters were used in ethyl 2-ethoxy-1,2-dihydroquinoline-1-carboxylate promoted condensation with 2-azidoethyl glycosides of glucuronic and galacturonic acid. Reduction of the azido-function followed byN-acryloylation and removal of blocking groups with trifluoroacetic acid gave the target monomers. These were converted into neoglycoconjugates of copolymer type, potentially useful for immunochemical studies.On leave from the Indian Institute of Chemical Technology, Hyderabad, India.     

Cytoarchitectonic pattern of the hypothalamus in the crocodile,Gavialis gangeticus

Summary The hypothalamus of the crocodile, Gavialis gangeticus, was investigated to reveal the organization of various nuclear complexes and to suggest homologies. The hypothalamic nuclei of G. gangeticus are composed of magnocellular and parvocellular neuronal entities. In the magnocellular system the nucleus supraopticus is well developed, whereas the nucleus paraventricularis and nucleus retrochiasmaticus are represented by scattered somata. Application of cytoarchitectonic criteria permits the delineation of 24 distinct parvocellular nuclear complexes extending rostrocaudally from the anterior commissure to the level indicated by the median eminence and nucleus mamillaris; some are further divisible into subgroups. The nucleus of the preoptic recess appears to be a unique property of the crocodilian hypothalamus. The nucleus suprachiasmaticus possesses a wing-like ventrolateral expansion that protrudes along the lateral aspect of the optic nerve. The tuberal region displays an elaborate pattern of nuclei segregated by regional specializations of the neuropil. The nucleus hypothalamicus posterior occupies the periventricular zone, flanked laterally by the nucleus hypothalamicus dorsomedialis and nucleus arcuatus. Further laterally, extended subdivisions of the nucleus hypothalamicus lateralis contain neurons rich in Nissl substance; the specializations shown by these subdivisions, in comparison to the lateral cell groups in lizards and snakes, are suggestive of enhanced integrative functions. The conspicuous paraventricular organ is encircled by dorsal and ventral divisions of the nucleus of the paraventricular organ. The neurons of the nucleus subfornicalis and nucleus hypothalamicus medialis are few in number, but large in size. The general organization of the hypothalamus of G. gangeticus reveals a mosaic-like pattern with the constituent groups appearing as clusters of small and large neurons, arranged medially and laterally in a definitive manner and accompanied by extensive zones of neuropil in the subependymal and lateral zones of the hypothalamus. The median eminence is divisible into an anterior and a posterior region. The nuclear pattern in the crocodilian hypothalamus reveals a higher state of morphologic organization compared to the situation in lizards or snakes, and thus reflects an evolutionary trend in the avian direction.     

SEX AND SPECIATION: GENETIC ARCHITECTURE AND EVOLUTIONARY POTENTIAL OF SEXUAL VERSUS NONSEXUAL TRAITS IN THE SIBLING SPECIES OF THE DROSOPHILA MELANOGASTER COMPLEX

Phenotypic divergence in the male reproductive system (genitalia and gonads) between species of the Drosophila melanogaster complex and their hybrids was quantified to decipher the role of these traits in species differentiation and speciation. Internal as well as external, sexual and nonsexual traits were analyzed with respect to genetic variation and trait asymmetry between strains within species, genetic divergence between species, and dominance and asymmetry in species and hybrids. The variation between strains within species was significant among sexual traits, and only external traits were less asymmetric than internal ones, which suggests that sexual traits are not strongly constrained within species. Three main findings show that sexual traits are most divergent between species: (1) testis length and area, and the area of the posterior lobe of the genital arch (sexual traits) showed the highest proportion of variation between species; (2) linear discriminant functions with the highest components associated to sexual traits were better predictors of species membership; and (3) testis length and area revealed a departure from a linear relationship between members of the species group. Examination of interspecific hybrids showed that sexual traits had higher asymmetry in species hybrids than in the parental species and that sexual traits showed additivity or dominance whereas nonsexual traits showed overdominance (with the exception of malpighian tubules length). These results suggest that sexual traits have undergone more genetic changes and, as a result, tend to show higher divergence and stronger hybrid breakdown between species than nonsexual traits. We propose that sexual selection in the broad sense, affecting all aspects of sexuality, may be responsible for the diversified appearance of sexual traits among closely related species and that the genetic architecture underlying sexual traits may be more prone to disruption during the early stages of speciation.     

Location and clonal analysis of stem cells and their differentiated progeny in the human ocular surface.

We have analyzed the proliferative and differentiation potential of human ocular keratinocytes. Holoclones, meroclones, and paraclones, previously identified in skin, constitute also the proliferative compartment of the ocular epithelium. Ocular holoclones have the expected properties of stem cells, while transient amplifying cells have variable proliferative potential. Corneal stem cells are segregated in the limbus, while conjunctival stem cells are uniformly distributed in bulbar and forniceal conjunctiva. Conjunctival keratinocytes and goblet cells derive from a common bipotent progenitor. Goblet cells were found in cultures of transient amplifying cells, suggesting that commitment for goblet cell differentiation can occur late in the life of a single conjunctival clone. We found that conjunctival keratinocytes with high proliferative capacity give rise to goblet cells at least twice in their life and, more importantly, at rather precise times of their life history, namely at 45-50 cell doublings and at approximately 15 cell doublings before senescence. Thus, the decision of conjunctival keratinocytes to differentiate into goblet cells appears to be dependent upon an intrinsic "cell doubling clock. " These data open new perspectives in the surgical treatment of severe defects of the anterior ocular surface with autologous cultured conjunctival epithelium.     

Zinc resistance in Neurospora crassa

Three non-identical Zn-resistant strains of Neurospora crassa have been isolated. ZNR-1 and ZNR-2 strains were obtained after repeated subculturing of wild type N. crassa on Zn-containing agar media (8mM and 16mM), while ZNR-3 was isolated after mutagenesis with diethyl sulfate, followed by selection on Zn agar plates (16mM). All three ZNR strains showed two- to threefold resistance to Zn in liquid media when compared with the wild type. However, growth measured by hyphal elongation clearly distinguished between the resistant strains (ZNR-3>ZNR-2>ZNR-1wild). The ZNR-2 and ZNR-3 strains were also cross-resistant to Co, while ZNR-2 alone was cross-resistant to Cu. Both Mg and Fe reversed the growth inhibition caused by Zn; Mg by suppression of Zn uptake and Fe without affecting the same. Assay of catalase, iron-binding siderophores and glutathione in Zn toxicity revealed significant increases in catalase and glutathione levels in the ZNR-2 strain when compared with the wild type. Kinetics of Zn uptake by preformed mycelia showed a rapid initial phase of uptake followed by a slower phase. The rates of Zn uptake measured after leaching surface-bound metal with EDTA revealed that ZNR strains have significantly reduced Zn uptake rates when compared with the wild type. The overall data suggest a partial transport block for Zn uptake as the major mechanism for resistance in ZNR strains. Genetic analysis of ZNR strains showed that in the ZNR-3 strain the znr locus maps close to the mating type locus (mt) of N. crassa LG I, while that of ZNR-1 and ZNR-2 is linked to LG IV associated with chromosomal aberration.     

The effect of chronic chlorpromazine administration on monoamine levels in various regions of rat brain

The neuroleptic drug, chlorpromazine (CPZ) has been shown to exert its antipsychotic effect by blocking post synaptic dopamine receptors. However, its effect on steady state levels of monoamines is still in discrepancy. In the present study, CPZ (4 mg/kg body weight) was administered intraperitoneally to adult Wistar rats chronically for 75 days and the levels of norepinephrine (NE), dopamine (DA) and 5-hydroxytryptamine (5-HT) were assayed in various brain regions by high performance liquid chromatography (HPLC). After the experimental period body and brain weights were not statistically different from controls. NE and 5-HT levels were increased only in hippocampus by 15% (p<0.01) and 16% (p<0.01) respectively. DA levels were consistently increased in cortex by 39% (p<0.001), striatum-accumbens by 18% (p<0.01), hippocampus by 27% (p<0.01), hypothalamus by 34% (p<0.001), cerebellum by 36% (p<0.001) and brainstem by 40% (p<0.001) in CPZ treated rats compared to controls. The results suggest that chronic CPZ administration increases DA levels in almost all regions of brain and reflect the ability of CPZ to preferentially interfere with synaptic transmission mediated by DA in brain. It also suggests that this increase in DA might be responsible for certain side effects seen in patients after chronic CPZ treatment.     

Delivering multiple anticancer peptides as a single prodrug using lysyl-lysine as a facile linker.

A large 40-residue precursor peptide (propeptide 5) was synthesized by linking together four designed anticancer peptide analogs to the neuropeptides: vasoactive intestinal peptide, somatostatin, bombesin and substance P, using enzyme cleavable lysyl-lysine linkers. On incubation with the enzyme trypsin, propeptide 5 was cleaved in a sequence-specific manner at the lysyl-lysine residues in the linker to release the individual peptide fragments which were identified by LC-MS. Another precursor peptide (propeptide 5a), consisting of two of the peptide analogs linked through lysyl-lysine linker, was also preferentially cleaved at the Lys-Lys site on incubation with the enzyme trypsin. Propeptide 5 showed potent anticancer activity, both in vitro and in vivo, which was greater than that of the individual component peptides. The enhanced activity suggests that the propeptide is possibly cleaved in the biological system at the lysyl-lysine site to yield the individual peptide analogs, which together show a synergistic effect. On the basis of these experimental findings, it can be concluded that pairs of basic amino acids such as Lys-Lys can be used as facile linkers for delivering multiple biologically active peptides.     

Variation and evolution of male sex combs in Drosophila: nature of selection response and theories of genetic variation for sexual traits

We investigated the genetic architecture of variation in male sex comb bristle number, a rapidly evolving secondary sexual character of Drosophila. Twenty-four generations of divergent artificial selection for sex comb bristle number in a heterogeneous population of Drosophila melanogaster resulted in a significant response that was more pronounced in the direction of low bristle numbers. We observed a strong positive correlated response to selection in the corresponding female transverse bristle row. The correlated response in male abdominal and sternopleural bristle numbers, on the other hand, did not follow the same pattern as sex comb bristle number differences between selection lines. Relaxation-of-selection experiments along with mate choice and fecundity assays using the selection lines developed demonstrated the action of stabilizing selection on sex comb bristle number. Our results show (1) substantial genetic variation underlying sex comb bristle number variation; (2) a weak relationship between the sex comb and developmentally related, non-sex bristle systems; and (3) that sexual selection may be a driving force in sex comb evolution, indicating the potential of sex combs to diversify rapidly during population differentiation and speciation. We discuss the implications of these results for theories of genetic variation in display and nondisplay male sex traits.