An unusual type of cytokeratin filament in cells of a human cloacogenic carcinoma derived from the anorectal transition zone

Epithelia-derived tumors (carcinomas) can be distinguished from mesenchymally derived tumors by the presence of intermediate-sized filaments of the cytokeratin type, which usually coincides with the absence of other types of intermediate-sized filaments such as vimentin filaments. In the course of diagnostic examinations of human tumors, using immunofluorescence microscopy, we have come across a case of an unusual carcinoma (Primary tumor and lymph node metastasis) positively stained not only with cytokeratin antibodies but also with immunoglobulins present in vimentin antisera. Therefore, this tumor, a cloacogenic carcinoma apparently derived from the rectal-anal transitional region, has been examined in greater detail using both immunofluorescence microscopy and immuno-electron microscopy as well as gel electrophoretic analysis of cytoskeletal polypeptides from total tumor tissue and from microdissected nodules enriched in carcinoma cells. The unusual reaction of the carcinoma cells with immunoglobulins present in seven different (rabbit or guinea pig) antisera raised against vimentin, has been found to be diminished after absorption on purified cytokeratin or total epidermal cytoskeletal material, but not after absorption on purified vimentin. Gel electrophoretic analysis of tumor cytoskeletons showed an unusual complex pattern of cytokeratin polypeptides containing relatively large (Mr 68,000 and Mr 58,000) neutral-to-slightly basic cytokeratins, as are typically found in epidermis and other stratified squamous epithelia, as well as several smaller acidic cytokeratins, including a Mr 40,000 polypeptide found in certain nonstratified epithelial such as colon and small intestine. Total tumor also showed the inclusion of some vimentin which, however, was significantly decreased in analysis of excised carcinoma nodules. Examining antibody binding to polypeptides separated by gel electrophoresis and blotted on nitrocellulose paper, we have found that antisera raised against vimentin contained not only vimentin antibodies but also immunoglobulins which specifically bound to the largest cytokeratin component. We conclude that the unusual reaction of immunoglobulins present in vimentin antisera with cytokeratin filament bundles does not represent specific binding to vimentin in these carcinoma cells, but is due to a component obviously widespread in vimentin antisera which binds specifically to a cytokeratin present in this type of tumor but not in most other carcinomas. It is proposed that use is made in diagnostic examinations of vimentin antisera or affinity-purified vimentin antibodies that have been pre-absorbed on cytokeratin protein, in order to eliminate such disturbing reactions.     

Photoperiod and Temperature Regulation of Floral Initiation and Anthesis in Soya Bean

Floral development includes initiation of floral primordia andsubsequent anthesis as discrete events, even though in manyinvestigations only anthesis is considered. For ‘Ransom’soya bean [Glycine max (L.) Merrill] grown at day/night temperaturesof 18/14, 22/18, 26/22, 30/26, and 34/30 °C and exposedto photoperiods of 10, 12, 14, 15, and 16 h, time of anthesisranged from less than 21 days after exposure at the shorterphotoperiods and warmer temperatures to more than 60 days atlonger photoperiods and cooler temperatures. For all temperatureregimes, however, floral primordia were initiated under shorterphotopenods within 3 to 5 days after exposure and after notmore than 7 to 10 days exposure to longer photoperiods. Onceinitiation had begun, time required for differentiation of individualfloral primordia and the duration of leaf initiation at shootapices increased with increasing length of photoperiod. Whileproduction of nodes ceased abruptly under photoperiods of 10and 12 h, new nodes continued to be formed concurrently withinitiation of axillary floral primordia under photoperiods of14, 15 and 16 h. The vegetative condition at the main stem shootapex was prolonged under the three longer photoperiods and issuggestive of the existence of an intermediate apex under theseconditions. The results indicate that initiation and anthesisare controlled independently rather than collectively by photoperiod,and that floral initiation consists of two independent steps—onefor the first-initiated flower in an axil of a main stem leafand a second for transformation of the terminal shoot apex fromthe vegetative to reproductive condition. Apical meristem, intermediate apex, floral initiation, anthesis, photoinduction, Glycine max(L.) Merrill, soya bean, photoperiod, temperature     

Microtubule assembly and disassembly at alkaline pH

Although it is now apparent that the intracellular pH may rise considerably above neutrality under physiological conditions, information on the effect of alkaline pH on microtubule assembly and disassembly is still quite fragmentay. We have studied the assembly/disassembly of bovine brain microtubule protein at alkaline pH in vitro. When microtubules are assembled to a new steady state at pH less than 7 and pH is then made more alkaline, they undergo a rapid disassembly to a new steady state. This disassembly is reversed by acidification. The degree of disassembly is determined largely by the pH- dependence of the critical concentration, which increases five to eight times, from pH 7 to 8. A fraction of assembly-incompetent tubulin is identified that increases with pH, but its incompetency is largely reversed with acidification. Measurements of microtubule lengths are used to indicate that disassembly occurs by uniform shortening of microtubules. A comparison of shortening by alkalinization with dilution suggests that the intrinsic rate of disassembly is accelerated by increasing pH. The capacity for initiating assembly is progressively lost with incubation at alkaline pH (although some protection is afforded by sulfhydryl-reducing agents). However, direct assembly from depolymerized mixtures is possible at least up to pH 8.3, and the steady state achieved at these alkaline pH values is stable. Such preparations are readily disassembled by cold and podophyllotoxin (PLN). Disassembly induced by PLN is also markedly enhanced at alkaline pH, suggesting a corresponding enhancement of “treadmilling.” The implications of physiological events leading to alkaline shifts of pH for microtubule assembly/disassembly are discussed, particularly in the light of recent hypotheses regarding treadmilling and its role in controlling the distribution of microtubules in vivo.     

Stability analyses for estimating relative durability of quantitative resistance

Summary Experiments in which a series of host cultivars are inoculated in all combinations with a series of pathogen isolates have been used to detect specificity in the host resistance. A theoretical model of polygenic resistance involving both general and specific interactions with pathogen virulence was developed to test the abilities of statistical analyses to discriminate between host genotypes with different levels of general and specific resistance. Estimates of levels of specific resistance could be obtained in regressions of disease severity scores for each host cultivar X pathogen isolate combination vs. the virulence index of each isolate. If the virulence index was based on the mean disease severity induced by the isolate over all host cultivars, the slopes of the regression lines were correlated with the levels of specific resistance in host cultivars. If the virulence index was based on the disease severity induced by the isolate on a host cultivar with a minimum of specific resistance, the mean squares for deviations from the regression were correlated with the levels of specific resistance in host cultivars. A method was developed to consistently choose host cultivars with minimum specific resistance. The two regression analyses gave estimates of specificity in randomly generated, model genotypes of approximately equal accuracy, although the second method appeared to be more accurate when the numbers of loci controlling resistance and virulence were small. The best estimates of numbers of genes for specific resistance were obtained by calculating a rating based on mean disease severity, the mean square for deviation from the regression on the virulence index based on disease severity on the cultivar with minimum specific resistance and the slope of the regression on the virulence index based on the mean disease severity. The best estimates of proportions of resistance genes that were specific were obtained by calculating a rating based on the above deviation mean square and slope alone.Cooperative investigation of the U.S. Department of Agriculture, Agricultural Research Service and the North Carolina Agricultural Research Service. Journal Series Paper No. 8326 of the North Carolina Agricultural Research Service     

THE EFFECTS OF DIURNAL AND TIDAL PERIODICITIES IN THE NUMBERS AND ACTIVITIES OF HERRING GULLS LARUS ARGENTATUS IN A COLONY

The average numbers of Herring Gulls Larus argentatus present in a breeding colony on Walney Island, Cumbria, were found to vary with the tidal cycle but to remain effectively constant with time of day through the breeding season. An activity survey, based on 50 Herring Gulls observed at half-hourly intervals during March and April 1973, showed that sleep and rest varied inversely with each other with sleep increasing to 50 per cent at midday. After a peak in the proportion of gulls asleep four hours before low tide, sleeping progressively decreased until low tide; seemingly a result of resident gulls waking and remaining more alert as others left the colony in search of food. Preening was constant throughout the day and tide cycle. Other behaviours (mostly courtship and agonistic behaviour associated with territory defence) increased slightly during low tide and were more common early and late in the day. Night observations of the gulls' activities showed that there was a peak of sleeping between midnight and 02.00 hours. It is suggested that Herring Gulls have a bimodal diel sleep pattern.     

A Malaria Parasite, Plasmodium aurulentum sp. nov. from the Neotropical Forest Gecko Thecadactylus rapicaudus

SYNOPSIS. Plasmodium aurulentum sp. nov. from the neotropical forest gecko Thecadactylus rapicaudus in Panama is characterized by oval or round to lenticular gametocytes, 6–22 nuclei in crudely fan-shaped schizonts, and light golden pigment masses. A prominent, pinkish red-staining mass, present in older schizonts, disappears by the time schizonts reach maturation.     

Fine Structure and Life Cycle of Lankesteria clarki sp. n. (Sporozoa: Eugregarinida) Parasitic in the Mosquito Aedes sierrensis (Ludlow)

Lankesteria clarki sp. n. from the treehole mosquito, Aedes sierrensis (Ludlow) in California, is described. This species can be separated from the closely related ones, L. culicis (Ross) and L. barretti Vávra, by the totally intracellular nature of the trophozoite, the shape of the gamont, the position of the gamont nucleus and the structure and position of the residual body in the spore. Morphologic and ultrastructural investigations of gametogenesis and sporogony were conducted and the host relationship discussed. Lankesteria clarki was found in mosquitoes collected from all treeholes examined near Novato in Marin County, California and an examination of over 6,000 larvae of A. sierrensis showed an infection rate of 27.5%. The parasite is capable of destroying the midgut epithelial cells during its trophic stage and Malpighian tubule cells during gametogenesis and sporogony.     

Insect Neuromuscular Synapses

Miniature end-plate potentials were used in studying severalaspects of the neuromuscular systems in the cockroach femur.The similar sizes and time courses of miniatures associatedwith fast and slow type excitatory axons suggest that they employthe same transmitter. There is other evidence also indicatingthat the essential difference between these two excitatory systemsis in the number of packets of transmitter released per nerveimpulse rather than different transmitter substances. From theshapes of miniatures it was suspected that typical muscle fibersmight have a branching structure. This was confirmed by histologicalexamination, intracellular stimulation, and intracellular dyeinjection. That inhibitory transmission is quantal is indicatedby hyperpolarizing miniatures which occur at random time intervals.Inhibitory transmission can be made to fail and recover in astepwise manner by manipulating the Ca/Mg ratio. In studiesof toxins which affect transmitter release at vertebrate motorend-plates, botulinal toxin was found to be without effect ateither excitatory or inhibitory junctions in cockroach muscle.However, black widow spider venom acted as it does in vertebrates,promoting massive release of transmitters and then permanentinactivation of the junctions.     

Lek Social Organization and Reproductive Success in the Greater Prairie Chicken

Between 1964 and 1969, studies were made of Prairie Chickenreproductive behavior on a lek with a stable social organization.The social organization of this lek was altered by the removalof dominant males; three in 1970 and two in 1971. Social organizationand reproductive behavior were observed on this lek prior toand following removal of the dominant males. Removal of dominantmales disrupted the stable social organization on the lek. Totalnumber of aggressive interactions between males increased followingremoval of dominant males.No change was noted in the total numberof females visiting the lek in 1970 compared with the previous6-year period. Likewise, no change was noted in the number ofattempted copulations observed in 1970 compared with previousyears. However, the number of successful copulations was drasticallyreduced (90% fewer) during 1970 because of increased male-maleaggressive interactions on the lek. During 1971, fewer territorialmales were on the lek compared to previous years, and fewerfemales visited the lek and fewer successful copulations wererecorded than in previous years.Late nests of Prairie Chickensare less successful and contain fewer eggs compared with nestsbegun earlier. In some instances, the female social hierarchymay serve to delay onset of nesting, therefore reducing totalreproduction for the year.