The second annual meeting of the NSCORT in Gravitational Biology was held at Kansas State University on September 29-October 1, 1992. Symposium presentations at the meeting included ones on basic gravitational cellular and developmental biology, spaceflight hardware for biological studies, studies on Space Shuttle, and special talks on Space Station Freedom and on life support systems.
The alpha crystallins are cytosolic proteins that co-localize and co-purify
with actin-containing microfilaments. Affinity column chromatography
employing both covalently-coupled actin or alpha crystallin was used to
demonstrate specific and saturable binding of actin with alpha crystallin.
This conclusion was confirmed by direct visualization of alpha aggregates
bound to actin polymerized in vitro. The significance of this interaction
in relation to the functional properties of these two polypeptides will be
discussed.
The distribution of organelles within columella cells of sweet clover was
examined by transmission electron microscopy following growth under static
or clinorotating conditions. A developmentally conditioned polarity was
observed, with a proximal location of the nucleus and a distal accumulation
of the endoplasmic reticulum. This polarity was insensitive to
clinorotation. In contrast, clinorotation altered the location of
amyloplasts. Application of cytoskeletal poisons (colchicine, cytochalasin
D, taxol, and phalloidin), especially during clinorotation, had interesting
effects on the maintenance of columella cell polarity, with a profound
effect on the extent, location, and structure of the endoplasmic reticulum.
The site of cytoskeletal interactions with sedimenting amyloplasts is
thought to be the amyloplast envelope. An envelope fraction, having
over 17 polypeptides, was isolated using immobilized antibody technology,
and will provide a means of assessing the role of specific peptides in
cytoskeleton/amyloplast interactions.
1Mount Desert Island Biological Laboratory, Salsbury Cove, Maine
04672.
Fertilized eggs of Ilyanassa obsoleta Stimpson were collected
immediately after their deposition in egg capsules. Unopened egg capsules
then were affixed to glass slides, and incubated either statically
(controls) or on a clinostat (experimentals). After incubation for 9-14
days, hatching occurred sooner and in a higher percentage of clinostated
capsules than in controls. Embryos that hatched while undergoing clinostat
incubation were abnormal in morphology, whereas other embryos present in
non-hatched capsules in the same tubes appeared normal, as did embryos in
the control tubes. Although the results are compatible with a conclusion
that vector-averaged gravity in the experimental tubes caused the altered
development, some other aspects of how the incubations were done may have
contributed to the differences between the control and experimental
results.
Functional reconstitution of purified preparations of human epidermal
growth factor receptor (EGFR) requires dissociation of the protein from its
plasma membrane lipid environment. Solubilization of membrane proteins in
this manner requires the use of detergents, which are known to disrupt
plasma membrane lipid/protein interactions. We have investigated to the
ability of three nonionic detergents to solubilize the human EGFR
selectively, and have also analyzed the effect of these various treatments
on the intrinsic tyrosyl kinase activity of the receptor. The nonionic
detergent known as n-octyl glucoside (n-octyl ß-D-glucopyranoside) was
found to give the best combination of selectivity, yield, and maintenance
of enzymatic activity of the human EGFR.
1Division of Biology, NSCORT, Section of Virology and Oncology, Kansas
State University, Manhattan, Kansas 66506.
The molecular mechanism participating in the transport of newly synthesized
proteins from the cytoplasm to the nucleus in mammalian cells is poorly
understood. Recently, the nuclear localization signal sequences (NLS) of
many nuclear proteins have been identified, and most have been found to be
composed of a highly basic amino acid stretch. A genetic "subtractive" and
a biochemical "additive" approach were used in our studies to identify the
NLS's of the polyomavirus structural capsid proteins. An NLS was
identified at the N-terminus
(Ala1-Pro-Lys-Arg-Lys-Ser-Gly-Val-Ser-Lys-Cys11)
of the major capsid protein VP1 and at the C-terminus
(Glu307-Glu-Asp-Gly-Pro-Glu-Lys-Lys-Lys-Arg-Arg-Leu318)
of the VP2/VP3 minor capsid proteins.
Incubation of murine fibroblasts with orthovanadate, a global tyrosine
phosphatase inhibitor, was shown to confer a "pseudo-transformed" phenotype
with regard to cell morphology and growth characteristics. This alteration
was manifested by both an increasing refractile appearance of the cells,
consistent with many transformed cell lines, as well as an increase in
maximum cell density was attained. Despite the abrogation of cellular
tyrosine phosphatase activity, orthovanadate-treated cells remained
sensitive to the biological activity of a naturally occurring
sialoglycopeptide (SGP) cell surface proliferation inhibitor. The results
indicated that tyrosine phosphatase activity, inhibited by orthovanadate,
was not involved in the signal transduction pathway of the SGP.
The lung rudiment, isolated from mid-gestation (11 day) mouse embryos, can
undergo morphogenesis in organ culture. Observation of living rudiments,
in culture, reveals both growth and ongoing bronchiolar branching activity.
To detect proteoglycan (PG) biosynthesis, and deposition in the
extracellular matrix, were metabolically labeled with radioactive sulfate,
then fixed, embedded, sectioned and processed for autoradiography. The
sulfated glycosaminoglycan (GAG) types, composing the carbohydrate
component of the proteoglycans, were evaluated by selective GAG degradative
approaches that showed chondroitin sulfate PG principally associated with
the interstitial matrix, and haparan sulfate PG principally associated with
the basement membrane. Experiments using the proteoglycan biosynthesis
disrupter, ß-xyloside, suggest that when chondroitin sulfate PG
deposition into the ECM is perturbed, branching morphogenesis is
compromised.
The unifying hypothesis of the NSCORT in gravitational biology postulates
that the ECM and growth factors are key interrelated components of a
macromolecular regulatory system. The ECM is known to be important in
growth and branching morphogenesis of embryonic organs. Growth factors have
been detected in the developing embryo, and often the pattern of
localization is associated with areas undergoing epithelial-mesenchymal
interactions. Causal relationships between these components may be of
fundamental importance in control of branching morphogenesis.
1Mount Desert Island Biological Laboratory, Salsbury Cove, Maine 04672.
Morphogenesis of the clearnose skate, Raja eglanteria, was not
significantly inhibited as a result of 7 days of exposure to 1-2 mM
selenate in the sea water during Days 59-69 of embryonic development
(hatching would normally have occurred at 82±4 days of incubation).
Although corneal transparency appeared normal in the eye, preliminary
measurements of the thickness of Bowman's layer of the cornea suggested
that it was significantly thinner in the corneas of embryos exposed to 1-2
mM selenate. Selenate is an ion reported to inhibit sulfation of
glycosaminoglycans in connective tissue.
By the end of 10th nuclear cycle, the somatic nuclei of the
Drosophila embryo have migrated to the periphery of the egg.
Centrifugation of embryos did not result in the displacement of these
nuclei, since cytoskeletal elements anchor them to the cortex. But, mild
centrifugal forces displace the centrally located, nascent yolk nuclei. If
this increased sensitivity to hypergravity occurs before the beginning of
nuclear differentiation during cycle 8, when the nascent yolk and somatic
nuclei physically separate, then it would mark the earliest functional
difference between these two lineages.
This manuscript briefly reviews ground-based and flight experiments,
discusses how those experiments complement each other, and details how
those experiments lead us to speculate about the gravity-sensitive nature
of protein kinase C.
1Chief Scientist, Space Station Freedom and
2Manager, Pressurized Payload Accomodations, Space Station
Freedom, Office of Space Systems Development, NASA Headquarters,
Washington, DC 20546.
The advent of Space Station Freedom (SSF) will provide a permanent
laboratory in space with unparalleled opportunities to perform biological
research. As with any spacecraft there will also be limitations. It is
our intent to describe this space laboratory and present a picture of how
scientists will conduct research in this unique environment we call space.
SSF is an international venture which will continue to serve as a model for
other peaceful international efforts. It is hoped that as the human race
moves out from this planet back to the moon and then on to Mars that SSF
can serve as a successful example of how things can and should be done.
Populations of wading birds in Sedgwick County, Kansas, were studied in
1991 to determine the size and species composition of active colonies, and
to document the locations of major foraging areas. Two colonies were
active that year: a rural one of 60-70 pairs of Great Blue Herons (Ardea
herodius), and a larger colony in Wichita, which contained 530 nests of
five species. Local breeding populations of wading birds are declining.
Apparent high levels of mortality were evident in the mixed-species colony,
with more than 160 eggs and 60 dead chicks found on the ground and in
nests. Egg predation by American Crows (Corvus brachyrhynchos) also
was documented. Interspecific differences were noted in the compass
directions of foraging flights taken by birds from the mixed-species
colony, and in the types of habitats used. Wading birds were observed at
more than 100 foraging sites.
A two-year study of the herpetofauna of the F.B. and Rena G. Ross Natural
History Reservation was conducted. A list of the herpetofauna of the
Reservation was prepared, and notes were made on the natural histories of
these animals. Field observations by the author, as well as recorded
field observations in the Reservation files, were used to compile the
herpetofauna list. In all, 10 amphibian species and 27 reptilian speicies
were found to occur, consisting of 2 salamanders, 8 frogs, 4 turtles, 6
lizards, and 17 snakes.
In 1987, the Arkansas River flowed throughout its course in southwestern
Kansas during much of the year. Streamflow was reduced or absent in many
segments of the stream during the summer of 1988, which has been the
typical case in recent years. Quantitative surveys of the ichthyofauna
were conducted in 1987 and 1988 to assess the initial impacts of
temporarily restored streamflow on the fishes. A total of 25 species of
fishes was collected during this study, compared to 22 species reported in
earlier surveys. None of the four previously reported taxa protected in
Kansas was collected, but six distributional records within Kansas and
increased populations of several species of fishes were documented in
1987.
Three localized laminated limestones occurring near the top of the Hamilton
paleochannel sequence (Virgilian) contain the well-known Hamilton fossil
assemblage, a thanatocoenosis of terrestrial, marine, nonmarine, and
euryhaline forms including plants, invertebrates ,and vertebrates
exhibiting "skin preservation." Dissolution or disaggregation and sieving
of the three limestones and associated limestone and mudstone units, the
Hamilton Fossil-Lagerstätte proper, has produced the following
additions to the Hamilton quarry (main quarry) assemblage: 1) the
ostracodes Amphissites, Geisina, Gutschickia?, Pseudobythocypris,
and Whipplella?, 2) two foraminiferids, an encrusting form and
Globivalvulina, and 3) the shark Xenacanthus. Additional
taxa are well-known fossils abundant in rocks of this age. As in the
situtation of previously identified taxa from the Hamilton
Fossil-Lagerstätte, the listed forms lived in a range of
paleoenvironments from freshwater (Whippella?) to marine (e.g.,
Globivalvulina), reflecting the coastal, and probably estuarine,
setting of the Hamilton beds.
For the lower alcohols, the tendency to serve as hydrogen bond donors (HBD)
toward solutes is commonly expressed through various semiempirical scales
incorporating polarity and acidity parameters. In this report a simplified
model for such HBD acidity is proposed which is based upon molecular
orbital perturbation theory. Two first-order perturbation terms are
included in the Hamiltonian which carry over to the total energy function
for the highest occupied molecular orbital (HOMO) of the alkanol.
Quantitative tests using dipolarity and HBD acidity data support the
two-term formulation of the Hamiltonian.
The collection of two Western Cottonmouths (Agkistrodon piscivorus
leucostoma) from Cherokee County in 1991 is reported. The history of
this taxon in Kansas is discussed and previous records are discredited.
The collection of the latest specimens confirms the presence of this taxon
in extreme east-central Cherokee County.
Myxidium typhonius, a myxosporidian protozoan, is described from the
gall bladders of 13 species of anuran amphibians in Peru, Deapartamento de
Madre de Dios, Provincia Tambopata, Cusco Amazónico, Rio Madre de
Dios, ca. 15 km E Puerto Maldonado. The ellipsoidal spores of the parasite
are distinguished from those of other myxidia infecting anurans on the
basis of overall size, nature of the sutural line, number of longitudinal
ridges, number and features of transverse ridges, polar capsule size, coils
in polar filament, and identical left and right sides. Each spore of M.
typhonius has a longitudinal suture extending pole to pole, 2
longitudinal ridges, and 9 or 10 transverse ridges. Two nuclei are located
between the 2 polar capsules and each polar filament has 4 or 5 turns
(coils). Average measurements (and ranges) in microns of 325 spores from
the frontal view are: spore length 10.9 (9.8-12.2); spore width 7.2
(5.7-8.9); polar capsule length 3.8 (3.3-4.9); polar capsule width 3.6
(3.3-5.0). Myxidium typhonius appears to be a common parasite of
the gall bladder of anurans from Cusco Amazonico, Peru, with an average
prevalence of 85.5%.
Increasing levels of atmospheric carbon dioxide suggest increasing
temperatures over the earth, all other causes of climate being constant.
This paper reviews changes to Kansas climate perceived by the mid-21st
century. Annual temperatures are expected to warm by some 9°F, with
precipitation remaining about the same as today, although perhaps
distributed somewhat differently during the year. The warming would cut
heating degree days by about 40%, increase cooling degree days by an
additional 170% (changing costs accordingly), and increase growing
degree days by about 50%. The latter change implies a faster growth rate
for field crops, but also implies increased moisture stress. A corn yield
model suggests that the warmer climate would decrease corn yields in
eastern Kansas by about 19%.
In this series of experiments, the effect of selenium (IV) oxide
(SeO2) on aromatic methyl groups was investigated. The products
obtained when 2-picoline, 4-picoline and 8-methylquinoline were oxidized by
selenium dioxide were 2-pyridine carboxylic acid, 4-pyridine carboxylic
acid and 8-quinoline aldehyde, respectively. The yield corresponding to
each product above was 50%, 77%, and 49%, respectively. Other compounds,
such as 2, 6-lutidine, 2-methylquinoline and 4-methylquinoline were also
studied. The products were 2, 6-pyridine dicarboxylic acid, 2-quinoline
aldehyde and 4-quinoline aldehyde. Additionally, the compounds which did
not react with SeO2 were toluene, 2-nitrotoluene, 2,
4-dinitrotoluene, 2-chlorotoluene and 4-chlorotoluene. The mechanism for
reactions involving selenium (IV) oxide is discussed in detail. The
products were analyzed qualitatively by using FTIR, NMR, and other physical
and chemical methods.
Wetapolystoma almae n. gen., n. sp. (Monogenea: Polystomatidae) is
described from the urinary bladder of Bufo typhonius (Linnaeus,
1758) collected in Cusco Amazonico, Peru. Its large terminal muscular oral
sucker, voluminous uterus, and long ovary with a straight fore-end
distinguish it from the other 3 polystomatid genera (Polystoma,
Mesopolystoma, and Riojatrema) known to inhabit South American
amphibians. The opistphaptor is wider than the body and armed with one
large pair of anchors with multiple striations and scleritized accessory
pieces concentrated at the base of the hook. There are eight hooks
surrounding the genital pore. The long and coiled uterus is filled with
many eggs, exhibiting ovoviviparous reproduction, the perfect adaptation to
a terrestrial host, a toad. Wetapolystoma almae is the perfect
polystome described from B. typhonius from Peru and the second
polystomatid genus from B. typhonius from South America. It is the
second polystomatid genus and species from Peru and the fourth genus known
to occur in South American amphibians.
Age structure and reproductive activity of beavers in Kansas were
determined by examining carcasses collected from trappers. Kits and
yearlings comprised 49% of the sample, and none of these were
reproductively active. The mean number of embryos and/or placental scars
in the uteri of female beavers older than yearlings was 2.9, and the litter
size varied with age. Data produced by this study indicated that the
beaver population sampled was not overexploited.
Mortality profiles of Mammuthus imperator (Leidy) were derived from
samples of isolated and socketed teeth recovered from the Rushville (Sh-3)
and Gordon (Sh-5) Quarries of Sheridan County, Nebraska. The Sh-3
mortality profile (MNI=11) is dominated by adult and old individuals and
closely corresponds to selective mortality of Loxodonta africana
after extended periods of drought. The Sh-5 mortality profile (MNI=7)
corresponds to mortality profiles that reproduce a stable age distribution
of East African L. americana, and time-averaged assemblages and mass
death events of other North American mammoth samples.
2Division of Biology, NSCORT, Kansas State University,
Manhattan, Kansas 66506.
3Southwestern College, Winfield, Kansas 67156.
4Address correspondence to G.W. Conrad at the Manhattan, Kansas,
address.
2Laboratory of Molecular Virology, National Cancer Institute,
Bethesda, Maryland 20892.
2Division of Biology, NSCORT, Kansas State University,
Manhattan, Kansas 66506.
3Mote Marine Laboratory, 1600 Thompson Parkway, Sarasota,
Florida 34236.
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