939. STROBILANTHES GLUTINOSA

Strobilanthes glutinosa is described and illustrated. It is a winter‐flowering species native to the foothills of the northwestern Himalayas. Although frost sensitive, it appears to flourish as a cultivated plant in shaded places enjoying a mild moist climate.     

787. STROBILANTHES NUTANS

The history of the discovery and naming of Strobilanthes nutans (Nees) T. Anderson is provided. It is described and illustrated. Some of the problems in trying to subdivide the genus Strobilanthes are discussed in the context of highlighting the distinctive features of S. nutans including the distinctive cone‐like inflorescence, the white flowers and the subglobose incurved anthers. The introduction and cultivation of S. nutans as well as the possibilities of the introduction of other hardy species of Strobilanthes are discussed.     

975. STROBILANTHES HAMILTONIANA

Strobilanthes hamiltoniana is described and illustrated. It is named after Francis Buchanan‐Hamilton who discovered it in Assam in 1808. Over the years, it has been known under different names which reflect both taxonomic changes and the variability of the species. Wild forms from the Himalayan region are predominantly white‐ or pale lilac‐flowered, whereas cultivated and naturalized plants found elsewhere are pink‐flowered and nearly always sterile. Propagation is, therefore, by cuttings. It is widely cultivated in places with a tropical climate or where tropical conditions can be replicated. It has become widely naturalized and even invasive on some oceanic islands.     

Heterosis at Allozyme Loci under Inbreeding and Crossbreeding in PINUS ATTENUATA

The dependence of heterosis at isozyme loci on inbreeding and crossbreeding was studied in 10-yr-old trees of knobcone pine (Pinus attenuata Lemm.). Heterozygosity was determined at 24 polymorphic isozyme loci and related to the rate of vegetative growth and cone production. The inbreds, created by selfpollination, had 46% of the heterozygosity of their mothers; the crossbreds, created by interpopulation crossing, had 155% of the heterozygosity of their mothers. Within the crossbreds, heterozygosity was positively correlated with trunk growth, but negatively correlated with cone production. Results in the crossbreds, however, were strongly influenced by a few individuals that showed unusually slow growth, high reproduction and low heterozygosity. Without those individuals, there was no relationship of heterozygosity to either growth or reproduction.—Within the inbreds, heterozygosity was positively correlated with both trunk growth and cone production. Each locus that was heterozygous in the mothers was calculated to mark about 3% of the genome for identity by descent in the inbred progeny; the total proportion of the genome marked was between 10 and 11%. Using these estimates to relate heterozygosity to the inbreeding coefficient (F) gave estimates of inbreeding depression per unit of F that fell within the range of published values for conifers. The strength of heterosis found among the inbreds suggests that single-locus or multilocus overdominance should be exceedingly difficult to detect in natural populations of predominantly outcrossing species.     

HETEROZYGOSITY AND DEVELOPMENTAL STABILITY UNDER INBREEDING AND CROSSBREEDING IN PINUS ATTENUATA

The relationship between stability of annual trunk growth and heterozygosity at 24 polymorphic isozyme loci was studied in 10-year-old trees of knobcone pine (Pinus attenuata Lemm.) that were the products of contrasting systems of mating, self-, and interpopulation cross-pollination. Heterozygosity and variability of trunk growth were strongly related only when inbreds and crossbreds were compared; the crossbreds showed greater residual variability on an absolute scale, and greater responsiveness to climate on both absolute and relative scales. Within the inbreds there was no evidence of a relationship between heterozygosity and variability. Within the crossbreds, only one trait, a measure of relative trunk growth rate, showed a relationship with heterozygosity, and indicated greater variability of the more heterozygous trees. These results, and others in the literature, suggest that the relationship of heterozygosity to homeostasis for fitness components is neither simple nor monotonic; it varies between scales of measurement, genetic backgrounds, and environments.     

SEXUAL DIMORPHISM IN SPOTTED DOLPHINS (STENELLA ATTENUATA) IN THE EASTERN TROPICAL PACIFIC OCEAN

We analyzed 4 meristic and 32 morphometric cranial traits of 612 adult specimens of spotted dolphins ( Stenella attenuata ) from the eastern tropical Pacific Ocean for sexual dimorphism. Eleven 5" latitude/longitude blocks with five or more of each sex were assessed with a two-way ANOVA for sex and geographic differences. Interaction between these two factors was found for six measurements, suggesting that the degree of sexual dimorphism varies geographically for a few characters (although in no discernible geographic pattern). Sexual dimorphism was demonstrated for 23 of the 36 characters, with differences ranging from 0.00 to 5.88 percent. Females characteristically had a longer rostrum, while males generally had larger skulls overall. In terms of number of characters, the extent of sexual dimorphism demonstrated for skulls of spotted dolphins goes considerably beyond that shown for any other small delphine. A discriminant function involving a combination of 10 characters enabled us to identify correctly the sex of more than 75 percent of the specimens. A procedure for correcting specimen measurements is outlined that would enable an investigator to combine male and female specimens in geographic variation studies. A term (zwitter) is proposed for use when referring to specimens where measurements have been corrected to take into account differences between the sexes.     

REPRODUCTIVE MATURITY AND SEASONALITY OF MALE SPOTTED DOLPHINS, STENELLA ATTENUATA, IN THE EASTERN TROPICAL PACIFIC

We estimated age at attainment of sexual maturity and examined reproductive seasonality for male spotted dolphins, Stenella attenuata , from the eastern tropical Pacific Ocean. Maturity was determined by histological examination of testes. Average age at sexual maturation was 14.7 yr (the mean of two readers' age estimates). Testis and epididymis weight and seminiferous tubule diameters were reliable indicators of maturity, whereas age, length and color phase were less reliable. Seasonality was determined by changes in testis and epididymis weight, relative quantity of spermatids and spermatozoa, and lumen diameter, as well as an index of testis development (weight of the right testis and epididymis divided by length of the right testis). Testis and epididymis weights and index values peaked in July and August, midway between two predicted mating seasons for the northern offshore stock, but spermatozoa levels were elevated during the predicted breeding seasons.     

ULTRASTRUCTURE OF AUXILIARY AND GONIMOBLAST CELLS DURING CARPOSPOROPHYTE DEVELOPMENT IN FAUCHEOCOLAX ATTENUATA (RHODOPHYTA)1

The ultra structure of post-fertilization development in Faucheocolax attenuata Setch. is described. Following fertilization and transfer of the diploid nucleus to the auxiliary cell, four gonimoblast initials usually are produced of the multinucleate auxiliary cell. Gonimoblast initials originally are uninucleate but undergo karyokinesis to form multinudeate gonimoblast cells. Terminal or generative gonimoblast cells cleave successively to form lobes of incipient carpospores, with each group of spores differentiating synchronously. Portions of the initial generative gonimoblast cells, however, remain to resume karyokinesis and repeat the process of cleavage into carpospores. Axial gonimoblast cells are transformed into secretory cells, which produce mucilage. Generative gonimoblast cells and auxiliary cells are similar in cellular structure. Both contain typical red algal proplastids, some dictyosomes, cytoplasmic concentric membranes, and numerous small vesicles. In addition, dark staining spherical masses, occurring in the cytoplasm of all cell types, may represent dehydrated haploid chromatin. Large septal plugs interconnect gonimoblast cells and the auxiliary cell. These plugs are small when first formed but increase dramatically in size during carposporophyte development.     

Ectopic expression of von Hippel-Lindau tumor suppressor induces apoptosis in 786-O renal cell carcinoma cells and regresses tumor growth of 786-O cells in nude mouse

The von Hippel-Lindau (VHL) is a known tumor suppressor that binds to alpha-subunits of hypoxia-inducible factors and induces ubiquitin-mediated degradation of the protein in an oxygen-dependent manner. VHL is also involved in the regulation of tumor angiogenesis, glycolysis, cell cycle regulation, and apoptosis. In the present study, we showed that ectopic expression of VHL induces apoptosis in renal cell carcinoma 786-O cells which contain only the mutant VHL, evidenced by TUNEL assay and DAPI staining. Furthermore, biochemical studies indicated that expression of VHL in 786-O cells results in both PARP and CPP32 cleavage, suggesting that VHL-induced apoptosis in 786-O cells is caspase dependent. Moreover, we also observed that apoptosis induced by ectopic VHL expression was associated with up-regulation of p27 as well as Bax, implicating the roles of these two proteins in VHL-induced apoptosis. The up-regulation of p27 and Bax by VHL was specific since we did not detect any changes in the level of other apoptotic factors including Fas and Bcl2 by the expression of VHL. We next examined the effect of VHL expression on the tumor growth of 786-O renal cell carcinoma cells in nude mouse. The results showed that injection of Ad.VHL adenovirus regresses the tumor growth of 786-O cells in nude mouse. The analysis by TUNEL assay as well as DAPI staining of 786-O tumors injected with Ad.VHL showed clear evidence of apoptosis. These results suggest that ectopic VHL expression induces apoptotic response in 786-O VHL mutant cells both in vitro and in vivo.     

Vascular invasion in human breast cancer is correlated to T-->786C polymorphism of NOS3 gene.

BACKGROUND: Nitric oxide (NO) is a free radical known to be a major regulator of vascular tonus, to inhibit cell proliferation, induce apoptosis, and be a mediator of macrophage cytostatic and cytotoxic effects. Recently, NO synthesis has been reported to be elevated in different cancers and is expected to promote metastasis by maintaining a vasodilator tone in blood vessels in and around the tumour. Two different common genetic polymorphisms were found on endothelial NO synthase (NOS3) gene: Glu298Asp on exon 7 and T-->786C in the promoter region. PURPOSE: To evaluate the impact of the NOS3 polymorphisms on vascular invasion and metastasis in breast cancer patients. DESIGN: Two NOS3 gene polymorphisms (Glu298Asp and T-->786C) were genotyped in 71 patients operated for breast cancer and followed for 6-30 months (median 21). A control population of 91 age and sex matched tumour-free subjects was also genotyped for the same polymorphisms. RESULTS: The distribution of both polymorphisms was not different between cases and controls. In patients without vascular invasion, T allele frequency was significantly lower than in patients with vascular invasion (p=0.033). At the end of the follow-up, T allele frequency was found to be less frequent in the metastasis free group than normal population (0.51 vs 0.64; p=0.047). CONCLUSION: Our results suggest that T allele reduction at the NOS3 promoter region may reduce vascular invasion in breast cancer and consequently reduce metastatic spread and be a favorable prognostic factor. These results need further validation in larger studies.     

eNOS gene T-786C polymorphism modulates atorvastatin-induced increase in blood nitrite

Statins inhibit cholesterol synthesis and produce pleiotropic, cholesterol-independent effects including endothelial NO synthase (eNOS) stimulation and increased expression. However, a functional polymorphism in the promoter of the eNOS gene (T-786C) reduces its activity and could modulate the response to statins. Here, we examined whether this polymorphism modulates the effects of atorvastatin on the plasma levels of markers of NO formation and oxidative stress. We genotyped 200 healthy subjects for this polymorphism, and 15 subjects with the TT genotype and 15 with the CC genotype were selected to receive placebo or atorvastatin 10 mg/day po for 14 days. To assess NO bioavailability, the plasma concentrations of nitrate, nitrite, and cGMP and the whole blood nitrite concentrations were determined after placebo or atorvastatin using an ozone-based chemiluminescence assay and an enzyme immunoassay. Thiobarbituric acid-reactive species (TBA-RS) were measured in the plasma to assess oxidative stress. Atorvastatin decreased cholesterol concentrations independent of genotype. Whereas atorvastatin produced no significant changes in plasma nitrite, nitrate, or cGMP concentrations in both genotype groups, atorvastatin increased whole blood nitrite concentrations and decreased plasma TBA-RS concentrations in the CC (but not in the TT) genotype group. These findings suggest that the T-786C polymorphism modulates the effects of atorvastatin on NO bioavailability and oxidative stress.     

GEOGRAPHIC PATTERNS OF VARIATION IN OFFSHORE SPOTTED DOLPHINS (STENELLA ATTENUATA) OF THE EASTERN TROPICAL PACIFIC OCEAN

The spotted dolphin (Stenella attenuata), a species commonly associated with the tuna fishery, is widely distributed in the eastern tropical Pacific Ocean (ETP). Geographic patterns in morphological characteristics of this dolphin were evaluated for 36 skull measurements of 574 museum specimens allocated to 5° latitude-longitude blocks. A strong subdivision between northern and southern offshore populations was demonstrated using principal components, canonical variates and cluster (UPGMA and function-point) analyses of block means derived from standardized, sex-adjusted measurements. Classification functions derived from a stepwise multiple discriminant analysis correctly identified more than 87 percent of the specimens as being from either a northern or southern locality. These results support recognition of southern spotted dolphins as a separate management stock. Dolphins from a sample representing a far-western locality show similarities with those from the south. Patterns of geographic variation in morphology were evaluated using Mantel tests and matrix correlations; 17 of the 36 characters showed regional patterning and 18 exhibited local patterning. Morphological trends were also assessed with respect to 14 oceanographic measures that characterize environmental variability in the ETP. In general, clinal variation in morphology mirrors a north-south trend found in several environmental variables. Overall, the most striking associations between morphological and environmental variables involved Solar Insolation (Jan.). Sea surface temperatures also exhibited a strong association with morphological variables; the highest correspondence was found between width of the temporal fossa and sea surface temperature in July. In addition, the thickness of the oxygen minimum layer exhibited a pattern of variation that was statistically associated with 58 percent of the skull characters. The geographic patterns found for cranial morphology and environmental measures are strongly clinal and in general congruent, suggesting that oceanographic conditions in part determine trends in morphological characteristics of spotted dolphins.     

Hyperthermia Induces Apoptosis of 786-O Cells through Suppressing Ku80 Expression

Hyperthermia as an anticancer method has been paid increasing attention in recent years. Several studies have shown that hyperthermia can kill tumor cells by inducing apoptosis. However, the underlying molecular mechanisms of hyperthermia-induced apoptosis are largely unknown. To investigate the effects and molecular mechanism of hyperthermia on the apoptosis in renal carcinoma 786-O cells, we firstly examined apoptosis and Ku expression in 786-O cell line treated with heat exposure (42°C for 0-4 h). The results showed that hyperthermia induced apoptosis of 786-O cells, and suppressed significantly Ku80 expression, but not Ku70 expression. Next, we knock-down Ku80 in 786-O cells, generating stable cell line 786-O-shKu80, and detected apoptosis, cell survival and cell cycle distribution. Our data showed higher apoptotic rate and lower surviving fraction in the stable cell line 786-O-shKu80 compared with those in control cells, exposed to the same heat stress (42°C for 0-4 h). Moreover, the results also showed suppression of Ku80 led to G2/M phase arrest in the stable cell line 786-O-shKu80 following heat treatment. Together, these findings indicate that Ku80 may play an important role in hyperthermia-induced apoptosis and heat-sensitivity of renal carcinoma cells through influencing the cell cycle distribution.