Cytochemical index of the function of the afferent fiber

A correlation between the spike activity and ferrous deposits in the Ranvier nodes of afferent fibers has been revealed in the frog bladder. The degenerated fibers reveal neither deposits, nor spike activity. The native fibers reveal deposits in the presence of spike activity. When the spike activity is suppressed by procainamide the node affinity to ferrous ions is retained. It is suggested that positive cytochemical reactions in the nodes demonstrate the retention of the initial morpho-functional structure of afferent fibers.     

Reflexotherapeutic effects on biologically active points of the concha auriculae

By means of histochemical method, for revealing cholinergic nervous structures, and electron microscopy, innervation of biologically active points (BAP) and that of neutral areas of the rabbit ear skin has been studied, normal and after acu-, electro- and electroacupuncture. The BAP have more intensive vascularization and innervation, their specific feature is presence of well pronounced nervous fasciculi. The latter are formed by 6-10 fibers 1-6 mcm in diameter. The diameter of the fasciculi is within the limits 25-30 mcm up to 40-45 mcm. Under the electron microscopic investigation myelin and amyelin fibers are revealed in the nervous fasciculi. In the area of the epidermal basal layer and in the epidermis itself, single nerve terminals are found; they are considered as the point pain receptors. After acu-, electro- and electroacupuncture, intensity of the nervous fibers staining increases, thus demonstrating an increment of acetylcholine esterase activity. After insertion of acupuncture needles and after electrical irritation, the changes in the nervous and cellular elements in the BAP areas are studied electron microscopically. After the effects mentioned, mast cells situating in the BAP area become, as a rule, degranulated. After repeated electroacupuncture irritation of the BAP, an inflammatory focus appears with peculiarities specific for the given process. The reflexotherapeutic effect is supposed to be transferred via the nervous system. The mechanical irritation of the nerve fasciculi and the accompanying traumatization of the surrounding cellular elements initiate the mechanism of reflexotherapy.     

Prenatal development of the intrinsic nerves of the muscles of the human foot

Myelogenesis and blood supply of the intraorganic nerves have been studied in 4-6- and 7-9-month-old human fetuses. At first, the intramuscular nerves are presented as very thick fasciculi (the diameter is more than 90), thick (the diameter is from 50 up to 90) and single muddle neural fasciculi (the diameter is from 30 up to 50 mcm). The microcirculatory blood bed is formed at the expense of branches of the blood vessel-satellites and the blood vessels of the surrounding tissues and is carried out, without any interruption, along the whole extent. In 7-9-month-old fetuses the neural apparatus becomes more complex. The number of the middle neural fasciculi appear. On the background of fine neural fibers in the fasciculi a small part of the middle neural fibers appears, and in the musculus flexor digitorum brevis--single thick neural fibers. The intramuscular nerves have their own hemocirculatory bed presented by microvessels that are on the perineurium surface, in its bulck and among neural fibers.     

Localized deposition of lead phosphate precipitate in glycerinated rabbit psoas muscle during ATP-induced contraction

The location of ATP-ase sites within sarcomeres was studied by allowing glycerinated rabbit psoas muscle to shorten under load in a solution containing ATP and lead nitrate. The lead phosphate precipitate formed during the contraction is located mainly within the A-band of shortened sarcomeres. In fibers glycerinated at rest length, the heaviest precipitate deposits are located at the center of the A-band, roughly corresponding to the double overlap zone, with lesser precipitate concentrations at the A-l boundary contraction bands. In fibers glycerinated at stretched length, shortened sarcomeres with heavy contraction bands at the A-l boundary and patent H-bands have heavy precipitate deposits over the contraction bands and no localization to the center of the A-band. The precipitate is thus seen to be most concentrated at those regions where one would expect to find the best contact between thick and thin filaments. Physiological experiments were performed. Evidence is presented that lead ions can substitute for calcium in the contraction process, altering the time course of contraction.     

An analysis of the relationships between subthreshold electrical properties and excitability in skeletal muscle

Skeletal muscle activation requires action potential (AP) initiation followed by its sarcolemmal propagation and tubular excitation to trigger Ca(2+) release and contraction. Recent studies demonstrate that ion channels underlying the resting membrane conductance (G(M)) of fast-twitch mammalian muscle fibers are highly regulated during muscle activity. Thus, onset of activity reduces G(M), whereas prolonged activity can markedly elevate G(M). Although these observations implicate G(M) regulation in control of muscle excitability, classical theoretical studies in un-myelinated axons predict little influence of G(M) on membrane excitability. However, surface membrane morphologies differ markedly between un-myelinated axons and muscle fibers, predominantly because of the tubular (t)-system of muscle fibers. This study develops a linear circuit model of mammalian muscle fiber and uses this to assess the role of subthreshold electrical properties, including G(M) changes during muscle activity, for AP initiation, AP propagation, and t-system excitation. Experimental observations of frequency-dependent length constant and membrane-phase properties in fast-twitch rat fibers could only be replicated by models that included t-system luminal resistances. Having quantified these resistances, the resulting models showed enhanced conduction velocity of passive current flow also implicating elevated AP propagation velocity. Furthermore, the resistances filter passive currents such that higher frequency current components would determine sarcolemma AP conduction velocity, whereas lower frequency components excite t-system APs. Because G(M) modulation affects only the low-frequency membrane impedance, the G(M) changes in active muscle would predominantly affect neuromuscular transmission and low-frequency t-system excitation while exerting little influence on the high-frequency process of sarcolemmal AP propagation. This physiological role of G(M) regulation was increased by high Cl(-) permeability, as in muscle endplate regions, and by increased extracellular [K(+)], as observed in working muscle. Thus, reduced G(M) at the onset of exercise would enhance t-system excitation and neuromuscular transmission, whereas elevated G(M) after sustained activity would inhibit these processes and thereby accentuate muscle fatigue.     

Localization of the peptide Hydra morphogen in the pontine neurons of the human brain

Localization of neurons with peptide "Hydra morphogen" in the human pons has been investigated by means of immunocytochemical methods. Single neurons are found in the cranial part of the pons, in its grey substance, that immediately adjoins the lateral walls of the IV ventricles and in the nucleus tegmenti dorsalis. These neurons are round and polygonal with body diameter 22-25 mcm, they contain light brown precipitate of various density. The data obtained corresponds to general evolutionary regularities on distribution of biologically active peptides in the nervous system of vertebrate and invertebrate animals.     

DNA content and expression of genes related to cell cycling in developing Gossypium hirsutum (Malvaceae) fibers

The cell cycle in cotton (Gossypium hirsutum) fibers is poorly understood. The objective of this study was to evaluate the cell cycle status and DNA content in developing cotton fibers. The DNA content and cell cycle distribution in fiber and hypocotyl cells were determined by flow cytometry. Expression levels of minichrosomal maintenance protein (mcm), cyclin B, and a retinoblastoma-like protein (rb) genes were determined with real-time PCR in fibers and dividing and nondividing tissues. No endoreduplication occurred, nor did genome size or percentage of G1-phase nuclei differ between hypocotyls and fibers. Approximately 13 and 17% of fiber nuclei were in the S phase in 14 days after anthesis (d) fibers and 25 d fibers, respectively. The mcm and cyclin B were expressed at higher levels in fibers than in mature leaves, but expression levels in fibers were less than 15% of meristematic tissues. Rb was expressed in fibers at levels less than 50% of mature leaves or meristematic tissues. Based on an apparent increase in S-phase cells as fibers mature and the low level of expression of genes associated with cell cycle progression, we conclude that S-phase arrest occurs in developing cotton fiber.     

Characteristics of the lymphoid cells of rabbits]

By means of scanning electron microscopy it has been stated that the thymus gland, spleen, paratracheal lymph node, Peyer's patches and appendix in an intact rabbit of "Shinshilla" strain have T- and B-lymphocytes in different proportion. T-lymphocytes are seen as spherical cells with rough surface, with a few microprocesses, have 3.0 +/- 0.1 mcm--4.7 +/- 0.2 mcm in diameter depending on their location. B-lymphocytes are seen as spherical cells with a considerable amount of microvilli on their surface, have 2.9 +/- 0.1 mcm--4.4 +/- 0.1 mcm in diameter depending also on their location. In the thymus gland, cells with rough surface, a few microprocesses and crater-like hollows prevail. In the spleen and lymph node, most of the cells are of similar diameter and have microvilli on the surface. Lymphoid population in the Peyer's patch and appendix is represented by large cells with microprocesses.     

拽线法:一个构建系统发育树的新算法

这篇文章要讨论的拽线法（DL）是贪婪算法的一种。和Fitch—Margoliash（FM）一样,DL也是基于距离矩阵构建系统发育树,但是和FM算法相比,DL具有低复杂度、较高的容错性和准确度高的优点。当存在误差时,DL算法只是加大了不在同一个父节点下的基因序列的距离,但能够准确的判断序列的亲缘关系,进而得到完美的进化树拓扑结构;相比之下,FM算法让各个基因序列间的距离均摊了这种误差,从而有可能将本应该具有相同父节点的基因序列分到不同的分支。     

Formation of structuro-functional units of the microcirculatory bed of muscles in the postnatal period of the ontogenesis of the white rat

By means of biomicroscopy, histological methods and scanning electron microscopy topological relationships between the white rat m. cremaster microvessels and changes of their spatial organization have been studied in the postnatal development beginning from the 3d to the 14th week. In the terminal link of the microcirculatory bed zonal functional complexes of microvessels--myoangions are revealed. They represent rather autonomic and regularly repeating constructions of microvessels, having spatial regularity in respect to muscle fibers. With age the myoangion increases its stretchness from 303 +/- 14 in 3-week-old up to (70 +/- 3) X 10 mcm in 14-week-old animals. Capillary density in the m. cremaster during the development decreases from 354 +/- 12 up to 210 +/- 4 mcm, respectively. Distance between the longitudinal capillaries increases from 16.1 +/- 1.3 up to 31.2 +/- 1.8 mcm. Increase of transversal connections takes place in the capillary network, increasing number of the branching knots; this demonstrates certain complication of the nutritive link of the muscle microcirculatory bed. By the time of sexual maturation (6 weeks) there is a sharp decrease of the inflow and outflow coefficients. Higher meanings of the coefficient (0.70-0.74) are noted before sexual maturation period in comparison to that in mature animals (0.53-0.55).     

Structural changes in Mcm5 protein bypass Cdc7-Dbf4 function and reduce replication origin efficiency in Saccharomyces cerevisiae

Eukaryotic chromosomal replication is a complicated process with many origins firing at different efficiencies and times during S phase. Prereplication complexes are assembled on all origins in G(1) phase, and yet only a subset of complexes is activated during S phase by DDK (for Dbf4-dependent kinase) (Cdc7-Dbf4). The yeast mcm5-bob1 (P83L) mutation bypasses DDK but results in reduced intrinsic firing efficiency at 11 endogenous origins and at origins located on minichromosomes. Origin efficiency may result from Mcm5 protein assuming an altered conformation, as predicted from the atomic structure of an archaeal MCM (for minichromosome maintenance) homologue. Similarly, an intragenic mutation in a residue predicted to interact with P83L suppresses the mcm5-bob1 bypass phenotype. We propose DDK phosphorylation of the MCM complex normally results in a single, highly active conformation of Mcm5, whereas the mcm5-bob1 mutation produces a number of conformations, only one of which is permissive for origin activation. Random adoption of these alternate states by the mcm5-bob1 protein can explain both how origin firing occurs independently of DDK and why origin efficiency is reduced. Because similar mutations in mcm2 and mcm4 cannot bypass DDK, Mcm5 protein may be a unique Mcm protein that is the final target of DDK regulation.     

Spatial relationship and conformational changes between the cardiac glycoside site and beta-subunit oligosaccharides in sodium plus potassium activated adenosinetriphosphatase

(Na,K)-ATPase, the enzyme responsible for active transport of Na and K across the plasma membranes of animal cells, consists of a catalytic subunit (alpha) and a glycoprotein subunit (beta) with unknown function. We have determined the distance between fluorescent probes directed to specific sites on the alpha- and beta-subunits and ligand-induced changes in the fluorescence of a probe specifically attached to the beta-subunit. The cardiac glycoside site on the alpha-subunit was labeled with anthroylouabain [Fortes, P. A. G. (1977) Biochemistry 16, 531-540]. The oligosaccharides on the beta-subunit were labeled with lucifer yellow carbohydrazide [Lee, J. A., & Fortes, P. A. G. (1985) Biochemistry 24, 322-330]. Resonance energy transfer from anthroylouabain to lucifer yellow was measured by steady-state and time-resolved fluorescence spectroscopy. The distance between these probes was determined from the efficiency of energy transfer. The average distance between anthroylouabain and lucifer yellow was 47 A and was independent of the number of acceptor molecules attached to the beta-subunit. The measured distance corresponds to the distance between the cardiac glycoside site and the center of the labeled oligosaccharides on the beta-subunit within one alpha beta dimer. The distance was the same (47 A) when anthroylouabain was bound with ATP or Pi as phosphorylating ligands but increased to 49 A in the presence of vanadate. The change in average distance provides quantitative evidence of a conformational difference between the complexes of cardiac glycosides with (Na,K)-ATPase induced by phosphorylating ligands or by vanadate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Size of the lesioned portion of the retina after laser exposure (electron microscopic study)

Morphological pattern of the retina in the trauma focus and outside its borders has been studied electron microscopically after application of ruby laser monoimpulse radiation (wave length 0.69 mcm, energy 1.06-1.25 X 10(-4) J and impulse length 20 nc). The trauma focus is not limited only by the volcano-shaped inflation of the retina. The lesion of the photoreceptory ultrastructure and the disturbance of microcirculation in the vascular layer are defined in 3 zones investigated: at the distance 50, 100 and 1,000 mcm from the spot on the retina, visible in the light microscope. This phenomenon should be taken into consideration when choosing therapeutic doses of laser radiation, applied for treatment of various diseases of the eye.     

Roles of Trm9- and ALKBH8-like proteins in the formation of modified wobble uridines in Arabidopsis tRNA

Uridine at the wobble position of tRNA is usually modified, and modification is required for accurate and efficient protein translation. In eukaryotes, wobble uridines are modified into 5-methoxycarbonylmethyluridine (mcm(5)U), 5-carbamoylmethyluridine (ncm(5)U) or derivatives thereof. Here, we demonstrate, both by in vitro and in vivo studies, that the Arabidopsis thaliana methyltransferase AT1G31600, denoted by us AtTRM9, is responsible for the final step in mcm(5)U formation, thus representing a functional homologue of the Saccharomyces cerevisiae Trm9 protein. We also show that the enzymatic activity of AtTRM9 depends on either one of two closely related proteins, AtTRM112a and AtTRM112b. Moreover, we demonstrate that AT1G36310, denoted AtALKBH8, is required for hydroxylation of mcm(5)U to (S)-mchm(5)U in tRNA(Gly)(UCC), and has a function similar to the mammalian dioxygenase ALKBH8. Interestingly, atalkbh8 mutant plants displayed strongly increased levels of mcm(5)U, and also of mcm(5)Um, its 2'-O-ribose methylated derivative. This suggests that accumulated mcm(5)U is prone to further ribose methylation by a non-specialized mechanism, and may challenge the notion that the existence of mcm(5)U- and mcm(5)Um-containing forms of the selenocysteine-specific tRNA(Sec) in mammals reflects an important regulatory process. The present study reveals a role in for several hitherto uncharacterized Arabidopsis proteins in the formation of modified wobble uridines.     

Two mcm3 mutations affect different steps in the initiation of DNA replication

Mcm3 is a subunit of the hexameric MCM2-7 complex required for the initiation and elongation of DNA replication in eukaryotes. We have characterized two mutant alleles, mcm3-1 and mcm3-10, in Saccharomyces cerevisiae and showed that they are defective at different steps of the replication initiation process. Mcm3-10 contains a P118L substitution that compromises its interaction with Mcm5 and the recruitment of Mcm3 and Mcm7 to a replication origin. P118 is conserved between Mcm3, Mcm4, Mcm5, and Mcm7. An identical substitution of this conserved residue in Mcm5 (P83L of mcm5-bob1) strengthens the interaction between Mcm3 and Mcm5 and allows cells to enter S phase independent of Cdc7-Dbf4 kinase (Hardy, C. F., Dryga, O., Pahl, P. M. B., and Sclafani, R. A. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 3151-3155). Mcm3-1 contains a G246E mutation that diminishes the efficiency of replication initiation (Yan, H., Merchant, A. M., and Tye, B. K. (1993) Genes Dev. 7, 2149-2160) but not its interaction with Mcm5 or recruitment of the MCM2-7 complex to replication origin. These observations indicate that Mcm3-10 is defective in a step before, and Mcm3-1 is defective in a step after the recruitment of the MCM2-7 complex to replication origins.     

Ultrastructural organization of the surface of the cingulate cortex of the cerebrum in rats

In the cingulate cortex of rats the marginal glia is predominantly presented as fibrillar astrocytes, their bodies are situated immediately at the surface. Numerous axons, dendrites, synapses and myelinated fibers are often arranged near the very surface and are separated from it with only 1-2 thin processes of glial cells. Along the whole cortical surface one can see a limiting membrane--a layer of non-cellular substance, situating at the distance of 60-100 mcm from plasmalemmas of the marginal astrocytes. Using ruthenium red, it is possible to reveal the glycocalix layer on the surface of the limiting membrane, as well as cords of the electron opaque substance, that connect it with plasmolemma of the superficial astrocytes. Three types of the cingulate cortex surface are described in rats: superficial areas to which cells of the pia mater membrane adjoin; areas where cells of the pia mater membrane are situated at various distance from the cortical surface and areas of close adjoining of the right and left hemispheres of the cerebrum. Sometimes the cleft between the hemispheres is completely reduced, and narrow lamellar-like cells of the pia mater membrane are tightly inserted between the limiting membranes of both hemispheres or adjoin the blood vessel, situating between the hemispheres. At the surface numerous gap and desmosome-like junctions are observed. This is especially important at the border where the media are separated. At injection of neurotoxin 6-hydroxydopamine certain ultrastructural rearrangements are noted in cytoplasm of the marginal astrocytes, changes in the number and extension of intercellular junctions.(ABSTRACT TRUNCATED AT 250 WORDS)     

嗜酸乳杆菌产生的广谱抗菌肽AP311的分离和鉴定

分离、鉴定了乳酸菌素片生产菌嗜酸乳杆菌 (Lactobacillusacidophilus)产生的热稳定性抗菌多肽AP31 1。浓缩的AP31 1对革兰氏阳性菌和革兰氏阴性菌均有抑制作用。AP31 1对枯草杆菌蛋白酶、凝乳蛋白酶、胃蛋白酶、胰蛋白酶、蛋白酶K敏感。AP31 1的抑菌活性随pH升高而降低 ,在酸性pH(pH2～ 4)条件下可经 1 0 0℃加热 30min活性不变。正丁醇可使AP31 1沉淀 ,当沉淀溶解于pH2的酸溶液时 ,活性恢复。氯仿、甲苯、乙酸乙酯、丁酮等有机溶剂对AP31 1活性没有影响。基于AP31 1可被蛋白酶酶解 ,具有广谱的抑菌活性 ,认为这种生物活性物质是一种抗菌肽。     

Acetylcholine inhibition in rabbit sinoatrial node is prevented by pertussis toxin

The effects of acetylcholine (ACh) were examined on the naturally occurring slow action potentials (APs) of the isolated, organ-cultured, spontaneously beating sinoatrial (SA) node of the rabbit, in the presence or absence of pertussis toxin. The sensitivity of the SA-node preparations to ACh was not altered after 24 h incubation in organ culture medium. Activation of the muscarinic receptor hyperpolarized the cells and reduced the frequency of spontaneous activity at low concentrations (1 X 10(-6) and 3 X 10(-6) M), and completely abolished automaticity at higher concentrations (1 X 10(-5) M). However, stimulated activity was maintained. Increased concentrations (1 X 10(-4) M) of ACh completely abolished excitability. When the SA-node preparations were cultured in the presence of 0.5 micrograms/mL pertussis toxin, concentrations of ACh as high as 1 X 10(-4) M had no effect on the AP parameters and frequency of spontaneous activity. The results indicate that inactivation of G proteins by pertussis toxin caused inhibition of the ACh effects on the automaticity of the SA node. In addition, the blocking effect of ACh to the naturally occurring slow APs was also inhibited by pertussis toxin. We conclude that in the rabbit SA node, the effects of ACh on automaticity and on the slow channels are mediated by G protein.