Influence of the age of the recipient on the manifestation of the effect of allogeneic inhibition]

The expression of allogenic inhibition was studied when transplanting 10(5) cells of bone marrow of C57BL mice to the lethally irradiated recipients (CBA X C57BL) F1 of different age (2 to 11 months). In the control experiments the bone marrow cells at the same dose were introduced to the lethally irradiated syngenic (C57BL) mice. The most pronounced inhibition of the parental stem cells proliferation was registered in 2 months old recipients F1 (4.7 times), it was somewhat weakened in 3 months old and animal in 4--11 months old recipients (1.1 to 1.8 times). The thymectomy of adult recipients F1 did not eliminate the expression of allogenic inhibition.     

Participation of the C-terminal region of the D1-polypeptide in the first steps in the assembly of the Mn4Ca cluster of photosystem II

Amino acid residue D1-Asp(170) of the D1-polypeptide of photosystem II was previously shown to be implicated in the binding and oxidation of the first manganese to be assembled into the Mn(4)Ca cluster of the oxygen-evolving complex (OEC). According to recent x-ray crystallographic structures of photosystem II, D1-Glu(333) is proposed to participate with D1-Asp(170) in the coordination of Mn4 of the OEC. Other residues in the C-terminal region of the D1-polypeptide are proposed to coordinate nearby manganese of the cluster. Site-directed replacements in Synechocystis sp. PCC 6803 at D1-His(332), D1-Glu(333), D1-Asp(342), D1-Ala(344), and D1-Ser(345) were examined with regard to their ability to influence the binding and oxidation of the first manganese in manganese-depleted photosystem II core complexes. Direct and indirect measurements reveal in all mutants, but most marked in D1-Glu(333) replaced by His, an impaired ability of Mn(2+) to reduce Y(Z)., indicating a reduced ability (elevated K(m)) compared with WT to bind and oxidize the first manganese of the OEC. The effect on the K(m) of these mutations is, however, considerably weaker than some of those constructed at D1-Asp(170) (replacement by Asn, Ala, and Ser). These observations imply that the C-terminal residues ultimately involved in manganese coordination contribute to the high affinity binding at D1-Asp(170) likely through electrostatic interactions. That these residues are far from D1-Asp(170) in the primary structure of the D1-polypeptide, imply that the C terminus of the D1-polypeptide is already close to its mature conformation at the first stages of assembly of the Mn(4)Ca cluster.     

兔主动脉前庭自律细胞与窦房结电生理特性的比较

为进一步阐明左心室流出道(主动脉前庭)自律细胞的特性,及其与窦房结细胞的异同,本实验利用常规的玻璃微电极细胞内记录技术,观察了一些离子通道阻断剂分别对离体兔窦房结起搏细胞与左心室流出道慢反应自律细胞的电生理特性的影响,重点探讨了这两种自律细胞的0期、4期去极离子流的异同。结果表明:(1)用1μmol/L维拉帕米(verapamil,VER)灌流后,窦房结及主动脉前庭自律细胞的动作电位幅值(APA)、0相最大除极速率(V_(max))、最大舒张电位(MDP)绝对值、舒张期除极速率(VDD)、自发放电频率(RPF)均明显下降,复极90％时间(APD_(90))延长(P<0.05)。(2)用180μmol/L氯化镍(NiCl_2)灌流,两自律细胞的VDD均明显下降;APA、V_(max)和RPF也显著降低,且窦房结细胞的APD_(90)明显延长。(3)给予2 mmol/L 4-氨基吡啶(4-AP)后,窦房结及主动脉前庭自律细胞的VDD均明显增快,MDP绝对值、APA和V_(max)显著下降,APD_(90)明显延长(P<0.05)。(4)给予2 mmol/L氯化铯(CsCl),两自律细胞的VDD及RPF均明显变慢。结果提示:(1)主动脉前庭自发慢反应电位的0相、4相去极离子流及复极离子流均与窦房结优势起搏细胞相似。(2)主动脉前庭起搏细胞Ca~(2+)内流为其0相主要去极离子流,复极过程主要由K~+外流引起,4相自动除极以K~+外流衰减为主,另外     

Characteristics of the receptive fields of neurons of the posterior temporal area of the cortex of the cat

In records of 219 single units in the posterotemporal cortical area (field 21) of nonanaesthetized cats, 51% of cells reacted to visual stimulation. The neurones had receptive fields (RFs) with central (0-10 degrees) or peripheral (10-52 degrees) localization in the visual field, their size increasing with eccentricity. Carting of RFs by a light bar scanning the visual field revealed a considerable variability of RFs shape, size and orientation in different cells. RFs sizes of the majority of recorded cells (100-1000 grad) were very large and exceeded the size of large RFs of neurones in the primary projection zone of the visual cortex.     

Factors influencing the properties of voltammetric carbon fibre electrodes: the importance of the pH of the medium used for the electrical treatment and of the resin coating of the fibres

Electrical treatment of resin-coated voltammetric carbon fibre electrodes with triangular voltage at low (1.1) pH resulted in electrodes almost insensitive to ascorbic acid (AA) and dihydroxyphenylacetic acid (DOPAC) while their response to 5-hydroxyindoleacetic acid (5-HIAA) was practically the same as after treatment in the usually employed pH 7.4 medium. Electrodes treated at high pH (12.0), on the other hand, were more sensitive to AA and DOPAC than those treated at pH 7.4 and less sensitive to 5-HIAA. Exposing resin-coated electrodes to the treatment media without electrical treatment was not sufficient to obtain the same results as with the application of the current. Electrodes without resin coating were sensitive to AA without electrical treatment while coated electrodes were not. Electrical treatment increased the sensitivity of non-coated electrodes and rendered coated electrodes even more sensitive than non-coated ones. Treatment of coated electrodes for a maximum sensitivity to 5-HIAA was found to require less time than to obtain maximum sensitivity to the other compounds. Present results suggest that it is possible to prepare selective voltammetric electrodes by choosing the right parameters for their electrical pretreatment.     

Protein markers in the microcyst of the posteroventral cochlear nucleus of the gerbil

Microcysts are most evident in the posteroventral and anteroventral cochlear nuclei (PVCN and AVCN) of the Mongolian gerbil. The origin and contents of the microcyst are not elucidated at present. The present study investigated the possible inclusions in the microcyst by employing immunocytochemical labeling to localize the existence of various protein markers. Thirty and 100 microm thick sections were used to substitute and reconstruct between 6 and 20 paraffin serial sections, respectively. In 30-microm-thick slice sections, immunoreactivity of glial fibrillary acidic protein (GFAP-IR), mitochondria inner membrane (MCA-In-IR), S-100 (S-100-IR), serotonin (5-HT-IR), myelin proteolipid protein (PLP-IR) and substance P (SP-IR) abutted on the perimeter of the microcyst. The immunolabeled SP-positive cells were adjacent to the evagination of the microcyst. In 100-microm-thick slice sections, immunoreactivity of nitric oxide synthase (NOS-IR) and somatostatin (SOM-IR) mainly precipitated as flocculent structures in the small to medium-sized microcysts. 5-HT-IR also precipitated as an elongated flocculent stalk adjacent to the large microcyst or randomly distributed in the neuropil. The findings suggest that GFAP, MCA-In, S-100, 5-HT, PLP, SP, NOS and SOM may be involved in modulating the physiological functions and maintaining micro-environmental homeostasis of the microcyst in the cochlear nucleus of the gerbil.     

Effect of desglycine-arginine vasopressin on the excitability of the command neurons of the defensive reflex in the edible snail

Perfusion of the snail (Helix lucorum L.) CNS with DG-AVP (concentration 10(-6) M) in the course of low frequency intracellular stimulation (2-4-minute interval) of the defensive reflex command neurons led to an increase in the excitability. It was expressed both in the reduction of the spike generation latency, in the increased number of spikes in response to fixed stimuli, and in the activation of pacemaker potentials. If DG-AVP was added to the medium during endoneuronal habituation, there was no increase in the excitability. It is supposed that modification of the neuronal excitability may be caused by the DG-AVP effect on the pacemaker mechanism.     

Effect of cortisol on the cellular proliferation and maturation in the cerebrum and the cerebellum of the rat: Importance of the age of the animals at the beginning of treatment]

Young rats were given either a single subcutaneous injection (1 mg at 0, 1, 4 or 8 days), or four consecutive daily injections (0.2 mg/day between 0 and 3 days; 0.4 mg/day between 4 and 7 days; 0.6 mg/day between 8 and 11 days) of cortisol acetate in order to test the influence of age on the action of corticosteroids on the biochemical maturation of the cerebrum and cerebellum in terms of their DNA, RNA, and protein contents. The results showed that: 1 The diminution of the DNA content at 35 days was greater in the cerebellum (- 16 to - 32%) than in the cerebrum (- 9 to 20%); the DNA content of the cerebrum was more affected by treatment at birth, whereas that of the cerebellum was more affected by the delayed treatments. Results were different when expressed in terms of reduction of the normal increase: the gain of DNA decreased more in the cerebrum (-70%) than in the cerebellum (-40%); but the most delayed treatment induced a greater effect in both organs. These abnormalities were not always accompanied by a significant decrease of the body weight. 2 Generally, the treatments led to an increase of the mean cell territory, expressed either in terms of decrease of the DNA concentration, or in terms of increase of the organ weight/DNA ratio. Moreover, the increase of the RNA/DNA and the protein/DNA ratios constituted an indication of an accelerated cellular maturation.     

Interrelationship of the mediator receptors in the terminals of the afferent nerves of the cornea of the eye]

In local application of substances with a mediator action (adrenaline, acetylcholine, histamine) and blockers of specific receptors (dihydroergotoxin, inderal, atropine and of subcutaneous dimedrol) together with dicaine it was possible to reveal a distinct presence in the cornea of the eye of the mediator receptors capable of influencing one another. Pharmacological analysis permitted a supposition to be made that there were two receptor systems in the sensory nerve endings of the cornea; these systems were in reciprocal inhibitory relations; due to this their regular influence on the processes of excitation and inhibition is effected in the sensory nerve ending.     

Effect of cold exposure on the concentration of triglyceride in the liver of the rat

The aim of the present study was to examine an effect of cold exposure on the concentration of triglycerides (TG) in the rat's liver. The rats were divided into the following groups: control, fed with oil, treated with hydrocortisone, fed with oil and treated with hydrocortisone, treated with noradrenaline. The rats exposed to cold were kept in wire cages (one rat in one cage) in the cold room at temperature +2 degrees C. They had free access to food (pellet diet for rodents) and water. In the control group the exposure to cold increased mildly (though significantly) the TG concentration after 1 and 3 h and had no effect after 2 and 24 h. It did not affect the concentration of plasma free fatty acids (FFA). At room temperature feeding with oil (2 ml/100 g of body weight) alone, and combined with hydrocortisone treatment (5 mg/100 g of body weight) as well as treatment with noradrenaline (0.1 mg/100 g of body weight) had no effect on the liver TG concentration, although the concentration of plasma FFA was increased. Exposure to cold for 3 h increased markedly the liver TG concentration in each of those groups. It is concluded that exposure to cold elicits a mechanism, which in the presence of elevated plasma FFA concentration induces accumulation of TG in the liver.     

Effect of piracetam on the habituation of the neuronal cholinoreceptor membrane of the edible snail

Piracetam is shown to modulate the habituation pattern of cholinoreceptor neuronal membrane in Helix lucorum. Piracetam (10(-2) M) intensified the reversible decrease of depolarization and spike discharge induced by the local microionophoretic rhythmical application of acetylcholine to the neuron soma. The ability of piracetam to intensify the habituation of neuronal cholinoreceptor membrane may serve as a model of its facilitating effect on the development of habituation of behavioural reactions. Piracetam (5.10(-2) M) was shown to induce a shift in the membrane potential towards depolarization in the majority of identified neurons studied.     

Analysis of changes in the electrical activity of the neocortex in dogs during the formation of the stereotype of food-procuring conditioned reflexes in them

A system of food-procuring conditioned reflexes (dynamic stereotype after I.P. Pavlov) was elaborated in dogs. In the interstimuli periods, 0.6 to 0.8 sec. prior the action of the conditioned stimulus, they exhibited a "state of expectancy" characterized by an increase of frequency (up to 80 per sec.) of potential oscillations of a small amplitude (20 to 30 mcv) and their pronounced sychroneity, predominantly in the anterior parts of the neocortex. The conditioned signal acting against such background produced specific reactions in the form of a limited number of bursts (3 to 4 in 0.5 sec.) of highfrequency (up to 100 per sec.) synchronized activity (HSA) of considerable amplitude (50 to 60 mcv) which always preceded conditioned reactions, being in certain temporal relations with them. A correlation-spectral analysis has shown that in the HSA period, electrical activity became considerably more regular; in the intensity spectra the extreme frequencies of the analyzed band (5 to 7 c/s and 90 to 100 c/s) became prominent, and in most cases high values of the coherence function were due to them. HSA reactions are regarded as a major link in the trigger mechanism of conditioned food-procuring reactions.     

Open junctions in the endothelium of the postcapillary venules of the diaphragm

We have previously established that approximately 30% of the endothelial junctions in the pericytic venules of the mouse diaphragm are open to a gap of approximately 30--60 A, and are fully permeated by hemeundecapeptide (H11P) (mol diam approximately 20 A). To estimate the size limit for molecules that can permeate these junctions, we have administered graded tracers intravenously and studied their behavior at the level of pericytic venules in bipolar microvascular fields (BMFs) in the mouse diaphragm. Horseradish peroxidase (HRP) (mol diam approximately 50 A) permeated only approximately 50% of the open junctions of the venular endothelium. Outflow through venular junctions appeared to be modest since the tracer remained restricted to the perivenular spaces. Hemoglobin (Hb, mol diam 64 x 55 x 50 A) permeated only a few (less than 5%), and ferritin (mol diam 110 A), practically none, of the endothelial junctions of the pericytic venules. The findings suggest that under normal conditions the size limit for permeant molecules for open venular junctions is approximately 60 A. Replicas of freeze-fracture preparations from appropriate regions in BMF showed that the intercellular junctions of the venular endothelium have the same organization as previously described for the corresponding segments of the microvasculature in the omentum and mesentery: discontinuous creases or grooves either free of or marked by few intramembrane particles only. Administration of histamine (topically or systemically) and 5-hydroxytryptamine (5-HT) (topically) resulted in typical focal separations of the endothelial junctions and intramural deposits of large tracer particles (carbon black) in the postcapillary venules.     

Ultrasonic evaluation of the corpus luteum of the mare

Two distinct luteal morphologies were observed in the ovaries of mares studied by daily ultrasound examinations. Luteal glands that formed after 48.5% of 95 ovulations were uniformly echogenic over 90 to 100 percent of the area of the image of the gland throughout the period of detectability. The remaining luteal structures (51.5%) exhibited a centrally located nonechogenic area. The nonechogenic area was first detected on day 0 (28%), day 1 (62%), day 2 (6%) or day 3 (4%) postovulation. Glands classified as centrally nonechogenic were echogenic over 80 to 100 percent of the area of the image of the gland on day 0; mean percentages of echogenic tissue decreased to 45 percent by day three then gradually increased to 95 percent before the glands became unidentifiable. The echogenic portion of the luteal glands of both morphologies had a bright echogenicity (gray-scale zone 4.5 to 5) on day 0. The echogenicity decreased (zones 3 to 3.5) by day 8 and was maintained at approximately that level until day 12. Mean gray-scale values tended to increase (zone 4 to 4.5) prior to the time the luteal glands became ultrasonically unidentifiable. These changes in grayscale values may have reflected changes in luteal hemodynamics. The nonechogenic area of centrally nonechogenic glands was attributed to clotted blood (corpus hemorrhagicum). The formation of a corpus hemorrhagicum was apparently not functionally important because it was present in only one half of the luteal glands. In addition, the mean length of time that the luteal gland was identifiable (17 days) or the mean length of the interovulatory interval (21 days) was not significantly different between the two luteal morphologies. Therefore, the hypothesis that the formation of a corpus hemorrhagicum is a necessary step in luteogenesis was not supported.     

Studies on the molecular mechanism of the translocation of estrogen receptor from the cytoplasm into the nucleus

A detailed study of the molecular mechanism of the translocation of estrogen receptor (ER) from the cytoplasm into the nucleus was undertaken in an in vitro system of porcine uterus. The capabilities of vero-ER . E (basic ER molecular bound with estradiol) (sedimentation coefficient 4.5S; Stokes radius 44 A) and the complexes ["5S" ER . E, (vero-ER . E) . (component A); "6S" ER . E, (vero-ER . E) . (component B)6; "8S" ER . E, (vero-ER . E) . (component B)6 . (component A)] with ER-binding factors (ERBFs) to translocate into the isolated nuclei were estimated by subtracting the amounts of ER adsorbed by the nuclear envelopes from those of ER bound to the whole nuclei. The results strongly supported our previous assumption that vero-ER . E translocates into the nuclei, and the complexes with ERBFs do not. The results suggested also that the binding site of vero-ER to ERBFs is required to be unoccupied in the process of the translocation of ER from the cytoplasm into the nucleus. The presence of a cytoplasmic factor (component C) which binds specifically with "5S" ER . E under low salt conditions was indicated. The complex, ("5S" ER . E) . (component C), was shown to possess relatively high affinity towards nuclear envelopes, but not to translocate into the nuclei.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of turn residues in directing the formation of the beta-sheet and in the stability of the beta-sheet

The designed peptide (denoted 20-mer, sequence VFITS(D)PGKTYTEV(D)PGOKILQ) has been shown to form a three-strand antiparallel beta-sheet. It is generally believed that the (D)Pro-Gly segment has the propensity to adopt a type II' beta-turn, thereby promoting the formation of this beta-sheet. Here, we replaced (D)Pro-Gly with Asp-Gly, which should favor a type I' turn, to examine the influence of different type of turns on the stability of the beta-sheet. Contrary to our expectation, the mutant peptide, denoted P6D, forms a five-residue type I turn plus a beta-bulge between the first two strands due to a one amino-acid frameshift in the hydrogen bonding network and side-chain inversion of the first beta-strand. In contrast, the same kind of substitution at (D)Pro-14 in the double mutant, denoted P6DP14D, does not yield the same effect. These observations suggest that the SDGK sequence disfavors the type I' conformation while the VDGO sequence favors a type I' turn, and that the frameshift in the first strand provides a way for the peptide to accommodate a disfavored turn sequence by protruding a bulge in the formation of the beta-hairpin. Thus, different types of turns can affect the stability of a beta-structure.     

Stimulation of the lateral hypothalamus provokes the initiation of robust long-term potentiation of the thalamo-cortical input to the barrel field of the adult, freely moving rat

Long-term potentiation in the thalamo-cortical input to the somatosensory cortex barrel field has been reported to be inducible in vitro only during a narrow critical period of the first postnatal week. Here we explored whether this is due to inability of adult synapses to express LTP or lack of appropriate conditions for LTP induction in slice preparations. We recorded thalamo-cortical field potentials (FPs) from the barrel field of chronically prepared adult rats. In the first series, several parameters of conditioning tetanization of thalamus (T) have been tried. Statistically significant LTP of 135-150% relative to the baseline was observed only in rare cases (3/18) so that the mean changes were not statistically significant. In the second series, five trains of 100 Hz stimulation of T were paired with a "reinforcing" stimulation of the lateral hypothalamus (LH). In most cases (9/13) thalamo-cortical FPs were potentiated. The mean post-tetanic amplitude was 238 +/- 42% (+/- SEM) relative to the baseline (n = 13). The potentiation persisted for > > 1 hr and typically even further increased when tested 24-48 hr later. LTP magnitude strongly correlated with the initial paired-pulse ratio (PPR, coefficient of correlation r = 0.98) so that LTP magnitude was larger (333 +/- 107, n = 6) in cases with PPR > 1.3. The mean PPR tended to decrease after LTP (from 2.05 to 1.65). Altogether the results suggest that LTP is inducible in the thalamo-cortical input to the barrel field of normal adult rats. The dependence of LTP magnitude upon the initial PPR suggests that inputs with low initial release probability undergo larger LTP. Together with the tendency to a decrease in the PPR this suggests an involvement of presynaptic mechanisms in the maintenance of neocortical LTP.     

The nature of the acid-volatile selenium in the liver of the male rat

1. The properties of rat liver acid-volatile selenium have been compared with those of H(2)Se and (CH(3))(2)Se. 2. In model experiments oxidation-sensitive H(2) (75)Se was trapped quantitatively under anaerobic conditions in 0.1m-AgNO(3), and (CH(3))(2) (75)Se was trapped quantitatively in 8m-HNO(3). The acid-labile selenium of a liver homogenate, and of a microsomal fraction, was found to behave quite unlike (CH(3))(2) (75)Se and in a manner indistinguishable from H(2) (75)Se. 3. It was concluded that the acid-volatile material is certainly not (CH(3))(2)Se and that it is probably H(2)Se. 4. The significance of these findings is discussed in relation to current knowledge about the metabolism and detoxication of selenium, and a scheme is proposed which incorporates this knowledge with recent observations on the interactions between trace amounts of selenium and tocopherol, and the production of acute selenium deficiency by Ag(+) in vitamin E-deficient rats.     

Influence of the structure of the lipid-water interface on the activity of hepatic lipase

Factors affecting the hydrolytic activity of purified rat hepatic lipase have been examined in mixed-monolayer systems. When nonsubstrate lipids [either egg sphingomyelin or beta-O-hexadecyl-gamma-O-(1-ocadec-9-enyl)-DL-phosphatidylcholine (OPPC-ether)] were used as inert matrices, hydrolytic activity for both triolein and dioleoylphosphatidylethanolamine was shown to decrease with increasing surface pressure (pi); negligible activity occurred at pi greater than or equal to 30 mN/m. Examination of the effect of introduction of cholesterol into either matrix containing 2 mol % triolein indicated that the mean molecular area decreased with increasing cholesterol and that, at pi = 24 mN/m, triolein was fully miscible in the sphingomyelin matrix at cholesterol concentrations less than or equal to 32.5 mol % and in the OPPC-ether matrix at cholesterol concentrations less than or equal to 49 mol %. Above these critical concentrations of cholesterol, the phase diagrams indicate transitions that suggest that triolein is forced out of the monolayer. Introduction of increasing amounts of cholesterol into either inert matrix increased the rate of hydrolysis of triolein by hepatic lipase, although by different degrees. There are at least two factors contributing to these effects: (1) condensation of the monolayer by cholesterol, thus increasing the total surface concentration of triolein at pi = 24 mN/m in the constant area surface balance, and (2) some change in triolein conformation and/or accessibility since at identical surface concentrations of triolein (8.7 +/- 0.1 pmol/cm2) and pi (24 mN/m) the rate of hydrolysis of triolein by hepatic lipase is 1.5-fold higher in the OPPC-ether matrix than in the egg sphingomyelin matrix.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in the electrical characteristics of bacterial cells in the disruption of the barrier function of the cytoplasmic membrane

A change in the electro-orientation spectrum followed by a change in the electric characteristics of bacterial cells had been measured when the barrier function of the cytoplasmic membrane was disordered under the action of damaging physical and chemical factors. Analysis of changes in the spectra at a frequency of 10(4) to 10(7) Hz allows one to estimate the degree of cell damage in quantitative terms. The paper presents the results of comparing three procedures for such an estimation: inoculation onto solid growth media; linear regression analysis of an electro-orientation spectrum; comparison of the electro-orientation effect at a high frequency (ca. 5 X 10(5) Hz) and at a medium frequency (ca. 10(4) Hz).