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31.
Thomas J. Contreras John F. Jemionek John E. French Virginia M. Hartwig Dane A. Walden 《Cryobiology》1980,17(3):243-251
Highly purified human granulocytes isolated from continuous flow centrifugation leukapheresis concentrates by counterflow centrifugation-elutriation were stored at 4 °C in concentrations of 6 × 106 to 1 × 107 granulocytes per milliliter for up to 14 days. The in vitro physiological function assays of phagocytosis, oxygen consumption associated with phagocytosis, bacterial growth inhibition, chemotaxis, and five enzyme analyses indicated good storage survival for up to 4 days. Stored granulocytes separated from other blood cells have greater storage stability than granulocytes stored as leukapheresis concentrates. After 14 days of storage a small percentage of granulocytes still maintained all physiological functions, with the exception of chemotaxis. Of the five enzymes assayed, only the enzyme activity of leucine aminopeptidase decreased significantly by the 14th day of storage. The storage stability of each physiological function assayed decreased as follows: bacterial growth inhibition (most stable), phagocytosis, oxygen consumption, and chemotaxis (least stable). 相似文献
32.
An apparently full-length complementary DNA copy of in vitro polyadenylated MS2 RNA was synthesized with avian myeloblastosis virus RNA-dependent DNA polymerase. After the MS2 RNA template was removed from the complementary DNA strand with T1 and pancreatic RNase digestion, the complementary DNA became a good template for the synthesis of double-stranded MS2 DNA with Escherichia coli DNA polymerase I. We then constructed molecular chimeras by inserting the double-stranded MS2 DNA into the PstI restriction endonuclease cleavage site of the E. coli plasmid pBR322 by means of the poly(dA)· poly(dT) tailing procedure. An E. coli transformant carrying a plasmid with a nearly full-length MS2 DNA insertion, called pMS2-7, was chosen for further study. Correlation between the restriction cleavage site map of pMS2-7 DNA and the cleavage map predicted from the primary structure of MS2 RNA, and nucleotide sequence analysis of the 5′ and 3′ end regions of the MS2 DNA insertion, showed that the entire MS2 RNA had been faithfully copied, and that, except for 14 nucleotides corresponding to the 5′-terminal sequence of MS2 RNA, the fulllength DNA copy of the viral genetic information had been inserted into the plasmid. Restriction endonuclease analysis of the chimera plasmid DNA also revealed the presence of an extra DNA insertion which was identified as the translocatable element IS13 (see following paper). 相似文献
33.
34.
Overlapping of the VP2-VP3 gene and the VP1 gene in the SV40 genome. 总被引:14,自引:0,他引:14
The nucleotide sequence of the SV40 Hind E fragment has been determined mainly by the partial chemical degradation procedure of Maxam and Gilbert (1977). The sequence of the strand with the same polarity as the late messenger RNA shows only one open reading frame for translation. Considering that VP3 corresponds to the carbosyl terminal part of VP2, and considering various evidence which indicates that the SV40 Hind E segment is part of the amino acid sequence of VP2-VP3. It continues clockwise in Hind K, where it terminates with a UAA signal. The latter is located 110 nucleotides beyond the initiation signal for the major structural protein VP1 (Fiers et al., 1975; Van de Voorde et al., 1976). Hence this small overlapping region of the genome codes for the synthesis of three different proteins in two different reading frames. The deduced amino acid sequence covers a major part of the vp3 poly peptide, and the amino acid composition is in good agreement with published values (Greenaway and Levine, 1973). 相似文献
35.
The product of the nitrogen fixation regulatory gene nfrX of Azotobacter vinelandii is functionally and structurally homologous to the uridylyltransferase encoded by glnD in enteric bacteria.
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A Contreras M Drummond A Bali G Blanco E Garcia G Bush C Kennedy M Merrick 《Journal of bacteriology》1991,173(24):7741-7749
We sequenced the nitrogen fixation regulatory gene nfrX from Azotobacter vinelandii, mutations in which cause a Nif- phenotype, and found that it encodes a 105-kDa protein (NfrX), the N terminus of which is highly homologous to that of the uridylyltransferase-uridylyl-removing enzyme encoded by glnD in Escherichia coli. In vivo complementation experiments demonstrate that the glnD and nfrX products are functionally interchangeable. A vinelandii nfrX thus appears to encode a uridylyltransferase-uridylyl-removing enzyme, and in this paper we report the first sequence of such a protein. The Nif- phenotype of nfrX mutants can be suppressed by a second mutation in a recently identified nifL-like gene immediately upstream of nifA in A. vinelandii. NifL mediates nif regulation in response to the N status in A. vinelandii, presumably by inhibiting NifA activator function as occurs in Klebsiella pneumoniae; thus, one role of NfrX is to modify, either directly or indirectly, the activity of the nifL product. 相似文献
36.
Tadimeti S. Rao Patricia C. Contreras Julie A. Cler Steve J. Mick Vickie M. Dilworth Smriti Iyengar Joseph B. Monahan Paul L. Wood 《Journal of neurochemistry》1991,56(3):890-897
Neurochemical interactions of tiletamine, a potent phencyclidine (PCP) receptor ligand, with the N-methyl-D-aspartate (NMDA)-coupled and -uncoupled PCP recognition sites were examined. Tiletamine potently displaced the binding of [3H]1-(2-thienyl)cyclohexylpiperidine with an IC50 of 79 nM without affecting sigma-, glycine, glutamate, kainate, quisqualate, or dopamine (DA) receptors. Like other PCP ligands acting via the NMDA-coupled PCP recognition sites, tiletamine decreased basal, harmaline-, and D-serine-mediated increases in cyclic cGMP levels and induced stereotypy and ataxia. Tiletamine was nearly five times more potent than PCP at inhibiting the binding of 3-hydroxy[3H]PCP to its high-affinity NMDA-uncoupled PCP recognition sites. However, following parenteral administration, dizocilpine maleate (MK-801), ketamine, PCP, dexoxadrol, and 1-(2-thienyl)cyclohexylpiperidine HCl, but not tiletamine, increased rat pyriform cortical DA metabolism and/or release, a response modulated by the NMDA-uncoupled PCP recognition sites. Pretreatment with tiletamine did not attenuate the MK-801-induced increases in rat pyriform cortical DA metabolism, a result suggesting that tiletamine is not a partial agonist of the NMDA-uncoupled PCP recognition sites in this region. However, following intracerebroventricular administration (100-500 micrograms/rat), tiletamine increased pyriform cortical DA metabolism with a bell-shaped dose-response curve. These data indicate a differential interaction of tiletamine with the NMDA-coupled and -uncoupled PCP recognition sites. The paradoxical effects of tiletamine suggest that tiletamine might activate receptor(s) or neuronal pathways of unknown pharmacology. 相似文献
37.
A. Illanes M. E. Zúñiga S. Contreras A. Guerrero 《Bioprocess and biosystems engineering》1992,7(5):199-204
A comprehensive methodology is presented for the design of reactors using immobilized enzymes as catalysts. The design is based on material balances and rate equations for enzyme action and decay and considers the effect of mass transfer limitations on the expression of enzyme activity. The enzymatic isomerization of glucose into fructose with a commercial immobilized glucose isomerase was selected as a case study. Results obtained are consistent with data obtained from existing high-fructose syrup plants. The methodology may be extended to other cases, provided sound expressions for enzyme action and decay are available and a simple flow pattern within the reactor might be assumed.List of Symbols
C kat/kg
specific activity of the catalyst
-
D m2/s
substrate diffusivity within the catalyst particle
-
Dr m
reactor diameter
-
d d
operating time of each reactor
-
E kat
initial enzyme activity
-
E
i kat
initial enzyme activity in each reactor
-
F m3/s
process flowrate
-
F
i m3/s
reactor feed flowrate at a given time
-
F
0 m3/s
initial feed flowrate to each reactor
-
H
number of enzyme half-lives used in the reactors
-
K mole/m3
equilibrium constant
-
K
S mole/m3
Michaelis constant for substrate
-
K
P mole/m3
Michaelis constant for product
-
K
m mole/m3
apparent Michaelis constant f(K, K
s, Kp, s0)
-
k mole/s · kat
reaction rate constant
-
k
d d–1
first-order thermal inactivation rate constant
-
L m
reactor height
-
L
r m
height of catalyst bed
-
N
R
number of reactors
-
P
i kg
catalyst weight in each reactor
-
p mole/m3
product concentration
-
R m
particle radius
-
R
P
ratio of minimum to maximum process flowrate
-
r m
distance to the center of the spherical particle
-
s mole/m3
substrate concentration
-
s
0i mole/m3
substrate concentration at reactor inlet
-
s
0 mole/m3
bulk substrate concentration
-
s mole/m3
apparent substrate concentration
-
T K
temperature
-
t d
time
-
t
i d
operating time for reactor i
-
t
s d
time elapsed between two successive charges of each reactor
-
V m3
reactor volumen
-
V
m mole/m3 s
maximum apparent reaction rate
-
V
p mole/m3 s
maximum reaction rate for product
-
V
R m3
actual volume of catalyst bed
-
V
r m3
calculated volume of catalyst bed
-
V
S mol/m3 s
maximum reaction rate for substrate
-
v mol/m3 s
initial reaction rate
-
v
i m/s
linear velocity
-
v
m mol/m3 s
apparent initial reaction rate f(Km, s,Vm)
-
X
substrate conversion
-
X
eq
substrate conversion at equilibrium
-
=s/K
dimensionless substrate concentration
-
0=s0/K
bulk dimensionless substrate concentration
-
eq=seq/K
dimensionless substrate concentration at equilibrium
-
local effectiveness factor
-
mean integrated effectiveness factor
-
Thiéle modulus
-
=r/R
dimensionless radius
-
s kg/m3
hydrated support density
-
substrate protection factor
-
s
residence time 相似文献
38.
Bone-marrow karyotypes of 68 specimens of the subterranean octodontid rodent genus Ctenomys from 16 different populations of north east Argentina and one from Paraguay have been studied. A surprising variety of chromosome numbers was found, ranging from 2n=42 to 2n=70. Some of the karyomorphs are clearly assigned to named species by topotypy: C. conoveris 2n=50, FN=56; C. argentinus, 2n=44, FN=54; C. perrensi, 2n=50, FN=84; C. dorbignyi, 2n=70, FN=84; C. roigi, 2n=48, FN=80; C. yolandae, 2n=50, FN=78. Four populations of Corrientes Province similar in morphology to C. perrensi were found to be polymorphic and polytypic; they maintain the same FN=84, but diploid numbers increase from 2n=54 to 2n=58 from SW to the NE, thus suggesting Robertsonian rearrangements. In the middle of this cline, a stable karyomorph of 2n=62, FN=84 was found in two different populations, suggesting to belong to an undescribed species. Another karyomorph of 2n=42, FN=76 found in Curuzú Laurel, Corrientes, may also prove to represent another undescribed species. One karyomorph of 2n=52, FN=74, and another of 2n=56, FN=78 from Paraná and Ubajaý (Entre Ríos Province, Argentina) respectively are close to C. rionegrensis. The relationships among these karyomorphs is considered in light of data on sperm morphology. The hypothesis is advanced that karyotypic rearrangements among the FN=84 group may be the result of Robertsonian repatterning from a 2n=70 original widespread form. Fixation of chromosomal variants is correlated with patchy distribution and small size of unstable demes, and may or may not have resulted in reproductive isolation. 相似文献
39.
The developmental profiles of four glycosidase enzymes (-D-glucosidase, -D-glucuronidase, -D-N-acetylglucosaminidase and -D-galactosidase) in the cochleas, cochlear nuclei and inferior colliculi of four strains of mice were investigated. The strains used were an audiogenically seizure-susceptible strain (DBA/2) and three non-susceptible strains (BALB/c, C3H/He and Swiss/A2G). The enzymic activities fell to varying degrees from 7 to 28 days of age. Two significant observations were made—-D-glucuronidase was low in the regions of the C3H/He strain, and -D-galactosidase was particularly low in the regions of the DBA/2 strain. The very low activity of -D-galactosidase in the DBA/2 mice is discussed in relation to the ganglioside patterns known to be present in these seizure-susceptible mice. Studies on the DNA contents of these auditory regions in the four strains showed no correlation with seizure sensitivity.This paper is dedicated to Dr. Derek Richter on his seventy-fifth birthday. 相似文献
40.
Summary Cellulolytic fungi were isolated from rotting leaves and tested for extra-cellular cellulase activities (CMCase, avicelase, cellobiase and xylanase). The effect of the proportion of the enzyme activities on the rate of degradation of leached beet cosette was observed using a range of supernatant fluids in appropriate combinations. At low cosette concentrations (1.5–3.0 g/l), avicelase and cellobiase were the rate limiting enzymes; avicelase in the initial stages of reaction and cellobiase after 6–8 hours, when cellobiose inhibition becomes important. A ratio of celiobiase to avicelase of approx 2.0 was established as appropriate. At higher substrate concentrations (10 g/l, 40 g/l) the best cellobiase to avicelase ratio was maintained and up to 40% hydrolysis was obtained in the 10 g/l incubation with 10 Uav/l and 20 Ucellob/l. At 100 g/l cosette concentration, substrate inhibition was observed. 相似文献