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991.
The N-terminal amino-acid sequence of the intrinsic membrane protein dipeptidyl peptidase IV (DP IV) was determined. The protein was isolated from pig kidney and solubilized by autolysis at pH 3.8. The first 34 amino acids were sequenced and indicated approximately 78% identity to the N-terminal sequence of rat liver DP IV. 相似文献
992.
In a series of bronchial and bladder carcinomas, p53 protein expression was examined. Samples from formalin-fixed, paraffin-embedded tissue (routine-treated) were compared with parallel samples of fresh tissue and tissue fixed in paraformaldehyde and ethanol. The expression of p53 was measured by immunofluorescence staining and dual parameter flow cytometry, with simultaneous monitoring of DNA content. For each tumor, p53 fluorescence with different fixatives was expressed relative to fresh tissue. The p53 fluorescence signals were on average brighter from routine-treated tissue than from fresh tissue. The tissue fixed in paraformaldehyde showed no difference from fresh tissue. In the ethanol-fixed tissue, however, fluorescence signals were reduced by nearly 70%, and the fraction of detectable p53 positive cells in tumor tissue was reduced by more than 50%. This loss of fluorescence was probably due to a leakage of the antigen from nucleus to cytoplasm. Pepsin treatment did not influence p53 fluorescence. Within the same tumor, the S-phase fraction in p53 positive cells was significantly higher than in p53 negative cells (13.1 +/- 2.0% vs. 6.5 +/- 0.8%). This pattern was not influenced by formalin fixation or pepsin treatment. Our study demonstrates that in measuring a nuclear antigen, tissue handling may influence the results, and care should be taken to optimize the preparation procedure. Using the antibody PAb 1801, p53 expression measured in archival material is not reduced as compared to fresh tissue. 相似文献
993.
M. Heinz R. Krattenmacher B. Hoffmann W. Clauss 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1991,161(4):363-370
Summary Transepithelial electrogenic Na+ transport (INa) was investigated in the coprodeum of 20-days-old chicken embryos in Ussing chambers. Short circuit current (Isc) and transepithelial resistance (Rt) were 14.7±4.8 A · cm-2 (n=12) and 0.53±0.09 k · cm-2 (n=12), respectively. INa was calculated from changes in Isc by substitution of mucosal Na+ by (N-methyl-d-glucamine) (NMDG). Isc inversed during Na+ removal, and INa was found to be 27.8±4.7 A · cm-2 (n=12). Amiloride (100 mol · l-1) inhibited only about 60% of INa. Analysis of Isc fluctuations revealed a Lorentzian component in the power density spectrum with a corner frequency of about 57 Hz. This component was not correlated to INa, and its origin is still unclear. Removal of mucosal Ca2+ increased INa about 2.5-fold due to an increase of the amiloride-insensitive component of INa in additionally investigated adult tissues. The results clearly show that this is due to a non-selective cation channel with an apparent order of selectivity Cs+>Na+=K+>Rb+>Li+. The Ca2+ concentration required to block 50% of the Isc was about 18 mol · l-1. The I
sc
Ca
could also be supressed by other divalent cations such as Mg2+ and Ba2+. Additionally, an INa-linked Lorentzian component occurred which dominated the control spectrum with a significantly higher corner frequency (about 88 Hz). The results indicate that Na+ absorption in the coprodeum of the chicken embryo is more complex than in adult hens. However, the Ca2+ sensitivity of INa is similar to comparable effects described for other epithelia. This possibly reflects the existence of two types of amiloride-insensitive apical cation channels as pathways for Na+ absorption, which may be involved to differing degrees in ontogenetic developments of nonselective channels to Na+-specific ion channels.Abbreviations
DPL
direct-linear-plot method
-
slope of the back-ground noise component
-
EGTA
ethylene glycol-bi(2-amino-ethylether)-N,N,N,N-tetraacetic acid
-
f
frequency
-
f
c
corner frequency of the Lorentzian noise component
-
G
t
transepithelial conductance
-
HEPES
N-hydroxyethylpiperazine-N-ethanesulfonic acid
-
I
sc
short-circuit current
-
I
Na
transepithelial sodium current
-
I
sc
Ca
Ca2+-sensitive short-circuit current
-
K
m
Ca
Michaelis-Menten constant for Ca2+
-
K
B
power density of the background noise component at f=1Hz
-
m
mucosal
-
NMDG
N-methyl-D-glucamine
-
R
t
transepithelial resistance
-
s
serosal
-
SEM
standard error of mean
-
S(f)
power density of the Lorentzian noise component
-
S
o
plateau value of the Lorentzian noise component 相似文献
994.
W H Schunck F Vogel B Gross E K?rgel S Mauersberger K K?pke C Gengnagel H G Müller 《European journal of cell biology》1991,55(2):336-345
cDNAs were cloned, sequenced and expressed which encode two different cytochrome P-450 forms of the alkane-assimilating yeast Candida maltosa, designated as P-450Cm1 and P-450Cm2. The amino acid sequences deduced were about 55% identical. Expression in Saccharomyces cerevisiae resulted in the formation of intact microsomal P-450 systems catalyzing the hydroxylation of n-hexadecane and lauric acid with significantly different substrate preferences. A massive proliferation of the endoplasmic reticulum was observed in the S. cerevisiae cells which produced P-450. Depending on the P-450 form expressed, distinctly organized stacks of paired membranes appeared and occupied considerable areas of the cytoplasm. As shown by immunoelectron microscopy for P-450Cm1, the protein expressed was highly concentrated within these newly formed membrane structures. 相似文献
995.
Klaus Hartfelder Heinz Rembold 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1991,160(6):617-620
Summary Juvenile hormone III content and ecdysteroid titer were analyzed for larval and pupal development of the stingless bee,Scaptotrigona postica depilis. Castespecific differences in juvenile hormone III content were detected at three developmental phases: at the transition from the fourth to the fifth larval stadium, in the spinning phase of the fifth larval stadium, and shortly after the imaginal moult. During the fifth larval stadium, juvenile hormone content closely reflects corpora allata activity. Juvenile hormone synthesis may thus be responsible for the elevated hormone titer in spinning-phase queen larvae, a phase of known sensitivity for induction of queen characters by exogenous juvenile hormone. For ecdysteroids, two phases of caste-specific differences were found: in the pre-pupal phase, and shortly after the imaginal moult. In both periods the titer in queens is distinctly higher compared to workers.Abbreviations
Im
imago 1 day after eclosion
-
L3, L4, L5
larval instars 3, 4, and 5
-
L5F1, L5F2
substages of feeding phase in fifth larval instar
-
L5S1, L5S2, L5S3
substages of spinning phase in fifth larval instar
-
PP1, PP2
substages of prepupal phase
-
Pw
white eyed pupa
-
Pp
pink eyed pupa
-
Pr
red eyed pupa
-
Pd
dark eyed pupa
-
Pdl, Pdm, Pdd
dark eyed pupa with progressive tanning of cuticle
-
RIA
radioimmunoassay 相似文献
996.
O Nilsson B W?ngberg A Wigander K Lundmark A Dahlstr?m H Ahlman A Bjartell R Ekman 《Peptides》1991,12(5):1077-1083
Delta sleep-inducing peptide (DSIP)-like immunoreactive (LI) material has been detected in nine different human pheochromocytoma tumors by immunocytochemistry. In primary tumors subjected to indirect immunofluorescence a variable number of tumor cells (25-75%) showed positive cytoplasmic labeling after incubation with DSIP antiserum. Tumor cells grown in culture were strongly labeled by the DSIP antiserum with DSIP-LI concentrated to cell bodies. Electron microscopic immunocytochemistry (immunogold labeling) of pheochromocytoma cells demonstrated DSIP-LI over the dense core of secretory granules. The presence of DSIP-LI in several HPLC fractions from conditioned culture media indicates secretion of DSIP-LI from cultured pheochromocytoma cells. The observations suggest that DSIP-LI is synthesized and stored in secretory granules before release. The different HPLC profiles from each of the tumors may reflect differences in processing or turnover of DSIP-LI in pheochromocytoma cells. 相似文献
997.
L P Panchenko O V Egorov I G Skripal' A S Ra?t E M Ivanova V F Zarytova V V Vlasov 《Mikrobiologicheekij zhurnal》1991,53(4):58-63
Stability of alkylating derivatives of decathymidylates protected on the 3'-terminal by cholesterol and phenazine residues has been studied in the process of their interaction with cells of Acholeplasma laidlawii PG-8. It is shown that the studied reagents are not split by nucleases of A. laidlawii PG-8 for the time necessary for alkylation of mycoplasma biopolymers. 相似文献
998.
A single X-ray irradiation of the rabbit hindlimbs in a dose of 0.24 C/kg evokes a decrease in fluorescence of the ANS probe bound with membranes of the sarcoplasmatic reticulum as a result of the decrease of binding sites, binding constant as well as the quantum output of the probe. A decrease in fluorescence of tryptophan residues of Ca-ATPase localized in membranes and attenuation of interaction of its SH-group with dithionitrobenzoic acid has been also observed at early postradiation terms (1 and 24 h). The obtained results evidence for structural rearrangements occurring in membranes of the sarcoplasmic reticulum under the effect of ionizing radiation. Changes in conformation of CA-ATPase molecules contribute much to this process. 相似文献
999.
V D Semenova V M Mikha?lover M L Zlochevski? K Lakhchev M Iu Beburov 《Molekuliarnaia genetika, mikrobiologiia i virusologiia》1991,(7):25-28
A gene for imidazole glycerophosphate dehydratase (HIS) has been selected from the library of Hansenula polymorpha genes by complementation of Escherichia coli hisB mutations or his3 mutation of Saccharomyces cerevisiae. Inactivation of the gene in Hansenula polymorpha strain 48V, as well as Leu(-)-Leu+ conversion of phenotype and arising of antibiotic G418 resistance resulted from transformation of the strain by the linear DNA molecules of the cloned HIS-gene with the in vitro inserted LEU2 gene from Saccharomices cerevisiae and KmR gene. The Southern hybridization analysis of the Leuf+His- G418R clones representing 1-2% of transformants population together with the DNA analysis of the monospore clones from the hybrid Leu+His-G418R transformant tetrad and tester strain have revealed the locus specific nature of the clones. 相似文献
1000.
Thin cardiac slices (100-200 microns) from newborn (1-14 days old) rat heart ventricles were used for patch clamp recordings. High resistance seals (10-50 GOhms) between patch-clamp pipettes and the membrane of cardiac cells as well as classical patch-clamp configurations can be achieved on this preparation without any enzymatic treatment of tissue. Resisting potential for cardiac cells measured in whole-cell configuration ranged between -30 and -65 mV. Averaged sodium currents and single inward rectifying potassium elementary currents recorded in cell-attached mode displayed basic features similar to those previously reported for isolated rat ventricular cells. Application of the method described here in cardiac electrophysiology will allow patch-clamp studies on heart cells without the complicated procedures of cell isolation. In addition, the uncertainty associated with enzyme treatment can be avoided. In future, this technique could be a new tool for studying electrophysiological properties of heart cells in situ. 相似文献