Localization of nitric oxide synthase in canine ileocolonic and pyloric sphincters

The distribution of neurons containing NADPH-diaphorase (NADPH-d) activity and nitric oxide synthase-like immunoreactivity (NOS-LI) in the canine pyloric and ileocolonic sphincters was studied. Cells within the myenteric and submucosal ganglia were positive for NADPH-d. These cells generally had the morphology of Dogiel type-I enteric neurons, however, there was some diversity in the morphology of NADPH-d-positive neurons in the myenteric plexus of the pylorus. Intramuscular ganglia were observed in both sphincters, and NADPH-d was found in a sub-population of neurons within these ganglia. Dual staining with an antiserum raised against nitric oxide synthase (NOS) demonstrated that almost all cells with NOS-LI were also NADPH-d positive. Varicose fibers within ganglia and within the circular and longitudinal muscle layers also possed NOS-LI and NADPH-d activity. Dual staining with anti-VIP antibodies showed that some of the NADPH-d-positive cells in the myenteric and submucosal ganglia also contained VIP-LI, but all VIP-LI-positive cells did not express NADPH-d activity. These data are consistent with recent physiological studies suggesting that nitric oxide serves as an inhibitory neurotransmitter in the pyloric and ileocolonic sphincters. The data also suggest that VIP is expressed in a sub-population of NADPH-d-positive neurons and may therefore act as a co-transmitter in enteric inhibitory neurotransmission to these specialized muscular regions.     

Heterochromatin accumulation,disposition and diversity in Gibasis karwinskyana (Commelinaceae)

C-banding differences within Gibasis karwinskyana (Roem & Schult.) Rohw. were reassessed using dual fluorochrome staining. Pronounced differences in C-band pattern between two subspecies with identical basic karyotypes were due to different chromosomal locations of AT-rich and GC-rich heterochromatin. The AT-rich component had an equilocal distribution in the karyotype and has evidently been accumulated at telomeres, as shown by its prevalence in supernumerary segments and B chromosomes. The GC-rich component also varied in amount, but was limited to nucleolus organizing regions (NORs) and centromeres. Centromeres and telomeres are suggested to constitute separate, although perhaps interdependent, centres of heterochromatin amplification. The possible role of nuclear architecture in determining the accumulation, distribution and spread of these sequences is discussed.Abbreviations H Hoechst 33258 - CMA chromomycin A3 - NOR nucleolus organizing region - SS supernumerary segment - Q quinacrine dihydrochloride - H⁺ H etc. indicate enhanced (+) and quenched (-) fluorescence with the stated fluorochrome by H.C. Macgregor     

DEVELOPMENT OF THE SEA-STAR, ASTERINA BATHERI GOTO

Using natural spawning and artificial fertilization, the entire process of development from eggs to juveniles was observed in the sea-star, Asterina batheri Goto.
The breeding season of this animal in Tsukumo Bay and Toyama Bay is estimated to be late summer. The spawned eggs are approximately 430 μm in diameter and float near the surface of sea water. They develop, through a wrinkled blastula stage by holoblastic. radial cleavage, into a pear-shaped brachiolaria bearing 3 blunt brachiolar arms. Metamorphosis takes place while the brachiolariae are swimming. Ten days after fertilization, metamorphosis is complete; the resulting juveniles are about 800 μm in diameter and colored pale brown with a green tint. They bear 2 pairs of tube-feet and a terminal tentacle in each arm.
Development of this species is thus of the direct type, and very similar in every respect to that of Asterina coronata japonica , which is closely related to the present species.     

Effect of Cullural Conditions on the Growth of Agrostemma githago Cells in Suspension Culture and the Concomitant Production of an Anti-Plant Virus Substance

An extract of cultured Agroxieinma githago L. cells was found to show potent inhibitory activity against plans virus infection. The effects of cultural conditions on the growth of the cell suspension and on the production of the inhibitor were examined. Since the production of substance was dependent on growth. experiments were made to improve growth. The optimum temperature was 26 to 30°C and optimum pH of the medium before autoclaving was between 5 and 7. In a medium of higher osmotic pressure, the water content of the cultured cells was lowered markedly. The growth rate in a small volume of the medium was higher than that in a larger volume at an early stage of the cultivation, but it was not changed by different inoculum sizes. The cells required thiamine and 2,4-D for growth but no other vitamins or growth regulators. The optimum level of 2,4-D was 0.1 mg/l. Higher sucrose concentration in the medium gave higher production of cell mass and of the inhibitor. However, 3% of sucrose was selected as the most economical concentration. For normal cell growth, the presence of both NH₄NO₃ and KNO₃ as nitrogen sources was required. The use of a single nitrogen source caused a long lag period or inhibition of the cell growth. KH₂PO₄ stimulated the growth when in was used in the level of 2.5 to 5 mM. The cell adhesion on the surface of the fermentor sometimes causes trouble in a large-scale cultivation. It was found that reducing the Ca²⁺ level in the medium prevented the cell adhesion and foaming remarkably. Based on the results obtained, a modified medium was established which was excellent for shortening the culture period and for efficient production of the anti-plant virus inhibitor.     

广州地区中华按蚊生理年龄的组成

中华按蚊是我国平原地区的传疟媒介,研究中华按蚊的生态学,对于疟疾的流行病学及预防均具有重要的意义。生理年龄及生殖营养环的研究,不但可以进一步认识中华按蚊在自然界的一些生态学问题,更重要的是有助于分析其传疟的作用,作为防制的依据。     

Oxygen Deficits,Clarity, and Eutrophication in some Madison Lakes

Oxygen deficits, rates of oxygen depletion, and Secchi disk measurements were the major indices examined in this study to determine trends in trophic conditions in Lakes Mendota, Monona, and Waubesa near Madison, Wisconsin. On the basis of available data, there is little that can be said reliably with respect to documenting eutrophication, particularly in Lake Mendota in the past several decades, that could not be attributed to normal variability. Better evidence for eutrophication comes from cores of some of the lakes. A comparison of oxygen deficits without simultaneous information on mean hypolimnetic temperatures and the lateness of ice out may provide a misleading interpretation of trophic variations over time.     

Cloning and expression profiling of testis-expressed microRNAs

Using a new small RNA cloning method, we identified 141 miRNAs from the mouse testis, of which 29 were novel. The 141 miRNAs were mapped onto all chromosomes except the Y chromosome and 2/3 of these miRNA genes exist as clusters. ∼ 70% of these miRNA genes were located in intronic or intergenic regions, whereas the remaining miRNAs were derived from exonic sequences. We further validated these cloned miRNAs by examining their expression in multiple mouse organs including developing testes and also in purified spermatogenic cells using semi-quantitative PCR analyses. Our expression profiling assays revealed that 60% of the testis-expressed miRNAs were ubiquitously expressed and the remaining are either preferentially (35%) or exclusively (5%) expressed in the testis. We also observed a lack of strand selection during testicular miRNA biogenesis, characterized by paired expression of both the 5′ strands and 3′ strands derived from the same precursor miRNAs. The present work identified numerous miRNAs preferentially or exclusively expressed in the testis, which would be interesting targets for further functional studies.