Modulation of the effects of mutations in the basic region of the OmpA signal peptide by the mature portion of the protein

Oligonucleotide-directed site-specific mutagenesis was used to study the structure-function relationship of the positively charged amino terminus of the Escherichia coli outer membrane protein OmpA signal peptide. Mutations were isolated which reduced the overall charge of the amino-terminal region from +2 (wild type) to +1, 0, and -1, as well as one mutation from Thr to Ser at position 4. DNA encoding the wild type and mutant OmpA signal peptides was then fused in-frame to DNA encoding the mature regions of Staphylococcus aureus nuclease A and TEM beta-lactamase. In the case of both the beta-lactamase and nuclease fusions, normal processing was no longer observed when the charge at the amino terminus was reduced to zero or made negative. Differences between the two hybrid proteins were observed in the case of the Thr to Ser mutation. As expected, this mutation had no effect on the beta-lactamase hybrid; however, the processing rate of the nuclease hybrid protein was reduced to nearly one-half. Furthermore, this effect was essentially reversed when a Lys residue at position 3 was deleted. A model is presented which explains the differing effects of a signal peptide mutation on the secretion of different hybrid proteins based on kinetic differences in the translocation of the nuclease and beta-lactamase proteins.     

Effects of the ablation of the nucleus pulposus on the vibrational behavior of the lumbosacral hinge

This study was designed to investigate the respective damping properties of the annulus fibrosus and nucleus pulposus of the intervertebral disc during propagation of vibration waves through the osteoligamento-muscular axis of the spine. The study was conducted on a 8-10 kg deeply anesthetized baboon. In the first surgical phase five accelerometers were implanted in the first sacral vertebra and on the anterior side of the four lower lumbar vertebrae. The bioinstrumented animal was placed in a restraining chair and exposed to narrow-bandwidth (0-100 Hz) 0.16 G RMS random vibration. Once data was recorded, the nuclei pulposi of the studied discs were removed by suction, the surrounding annuli remaining intact. The still deeply anesthetized animal was again exposed to the same 0-100 Hz, 0.16 G RMS vibration. Results were analyzed and their reproducibility was tested on three animals.     

CT与X线诊断肠梗阻的临床价值探讨

目的：探讨腹部X线与CT扫描鉴别急性肠梗阻的准确性,为临床诊断提供参考。方法：选取2011年8月-2013年8月我院收治的66例急性肠梗阻患者的临床资料进行回顾分析。所有患者均经手术活检或病理诊断证实为急性肠梗阻。术后患者均行腹部X线及CT扫描检查,对两种方法判断肠梗阻的发生、梗阻部位、类型及病因与手术病理结果进行比较,评价并分析两种检查方法的准确率。结果：66例肠梗阻患者中,X线检出率为89．39％,CT检出率为95．45％。X线诊断小肠梗阻准确率为72．10％,CT为86．05％;X线诊断结肠梗阻准确率为69．57％,CT为86．96％。X线诊断肿瘤准确率为69．57％,CT为86．96％;X线诊断肠粘连准确率为67．86％,CT为82．14％;X线诊断肠套叠准确率为60．00％,CT为80．00％;X线诊断单纯性机械性肠梗阻准确率为78．72％,CT为82．98％;X线诊断绞窄性肠梗准确率为73．68％,CT为78．95％。CT对肠梗阻部位、病因及类型的诊断准确率高于腹部X线片,差异具有统计学意义（P〈0．05）。结论：腹部X线与CT用于诊断急性肠梗阻具有较高的准确率,但CT对于肠梗阻部位、梗阻类型及梗阻病因的诊断优于X线片。     

CT 与X线诊断肠梗阻的临床价值探讨

目的：探讨腹部X 线与CT 扫描鉴别急性肠梗阻的准确性,为临床诊断提供参考。方法：选取2011年8 月-2013 年8 月我院 收治的66 例急性肠梗阻患者的临床资料进行回顾分析。所有患者均经手术活检或病理诊断证实为急性肠梗阻。术后患者均行腹 部X 线及CT扫描检查,对两种方法判断肠梗阻的发生、梗阻部位、类型及病因与手术病理结果进行比较,评价并分析两种检查 方法的准确率。结果：66 例肠梗阻患者中,X线检出率为89.39%,CT 检出率为95.45%。X 线诊断小肠梗阻准确率为72.10%,CT 为86.05%;X线诊断结肠梗阻准确率为69.57%,CT 为86.96%。X线诊断肿瘤准确率为69.57%,CT 为86.96%;X线诊断肠粘连 准确率为67.86%,CT 为82.14%;X 线诊断肠套叠准确率为60.00%,CT 为80.00%;X线诊断单纯性机械性肠梗阻准确率为78.72%,CT为82.98%;X线诊断绞窄性肠梗准确率为73.68%,CT 为78.95 %。CT 对肠梗阻部位、病因及类型的诊断准确率高于腹部X 线片,差异具有统计学意义(P<0.05)。结论：腹部X 线与CT 用于诊断急性肠梗阻具有较高的准确率,但CT 对于肠梗阻部位、梗阻类型及梗阻病因的诊断优于X线片。     

On the existence of parasympathetic motor nerves to the submaxillary gland of the dog

Duct pressure and salivary flow were recorded in submaxillary glands of anaesthetized dogs, to study whether parasympathetic stimulation caused effects referable to activity in myoepithelial cells. At fairly low frequency of stimulation, e.g. 3 Hz, the pressure curve had two distinct components, with initial steep and a secondary gradual rise. It resembled that obtained on sympathetic stimulation, where the first phase is ascribed to myoepithelial contraction, the second phase to secretion. When parasympathetic stimulation ceased, there was first a steep fall, then a more gradual decline of the pressure. The steep fall was of the same magnitude as the steep rise; both increased with the frequency of stimulation. The size of the initial fall was fairly independent of the pressure level from which it started. Such a steep fall did not occur subsequent to parasympathetic stimulation if the myoepithelial cells were already in a state of strong contraction caused by sympathetic impulses or bradykinin. The phase of steep fall was inferred to be due mainly to relaxation of contracted myoepithelial cells, the following decline to back-flow of fluid into the gland. The salivary flow rate was highest at the beginning of a period of parasympathetic stimulation, particularly if the duct system was well filled and the saliva thin. It was concluded that myoepithelial contraction had initially expelled saliva. A brief period of parasympathetic stimulation while a slow basal secretion at constant rate was going on was found to accelerate this flow, and afterwards there was a transient deceleration of the flow. Acceleration was attributed partly to myoepithelial contraction, mainly to superimposed secretion; retardation to myoepithelial relaxation. The effect appeared independently of the way in which the basal flow was evoked, and the retardation resembled that seen after sympathetic stimulation or bradykinin.     

The role of the methylation in the detoxication of arsenate in the rabbit

The biotransformation, tissue retention, intracellular binding and biokinetics of arsenic were studied in rabbits exposed to [74As]arsenate (0.4 mg As/kg body wt., i.v.). Inhibition of the methyltransferase activity by injection of periodate-oxidized adenosine (PAD) caused a marked decrease of the formation of [74As]dimethylarsinic acid (DMA), which gave rise to 1.5-4 times increased tissue levels of 74As. This is almost the same as reported for rabbits given arsenite in combination with PAD and was due to a rapid reduction of the arsenate to arsenite which bound to the tissues. Only about 30% of the arsenate given was excreted unchanged in the urine, indicating that a large part was reduced to AsIII. Thus the methylation to DMA seems to be almost as important for the detoxication following exposure to arsenate as that following exposure to arsenite. In the rabbits with normal methylating capacity 50-70% of the produced AsIII was methylated to DMA. The liver was the only organ in which DMA was present 1 h after the administration, indicating that this is the main site of the methylation. The DMA was rapidly cleared from all tissues except the thyroid.     

Determinants of the pH of the Golgi complex

The factors contributing to the establishment of the steady state Golgi pH (pH(G)) were studied in intact and permeabilized mammalian cells by fluorescence ratio imaging. Retrograde transport of the nontoxic B subunit of verotoxin 1 was used to deliver pH-sensitive probes to the Golgi complex. To evaluate whether counter-ion permeability limited the activity of the electrogenic V-ATPase, we determined the concentration of K(+) in the lumen of the Golgi using a null point titration method. The [K(+)] inside the Golgi was found to be close to that of the cytosol, and increasing its permeability had no effect on pH(G). Moreover, the capacity of the endogenous counter-ion permeability exceeded the rate of H(+) pumping, implying that the potential across the Golgi membrane is negligible and has little influence on pH(G). The V-ATPase does not reach thermodynamic equilibrium nor does it seem to be allosterically inactivated at the steady state pH(G). In fact, active H(+) pumping was detectable even below the resting pH(G). A steady state pH was attained when the rate of pumping was matched by the passive backflux of H(+) (equivalents) or "leak." The nature of this leak pathway was investigated in detail. Neither vesicular traffic nor H(+)/cation antiporters or symporters were found to contribute to the net loss of H(+) from the Golgi. Instead, the leak was sensitive to voltage changes and was inhibited by Zn(2+), resembling the H(+) conductive pathway of the plasma membrane. We conclude that a balance between an endogenous leak, which includes a conductive component, and the H(+) pump determines the pH at which the Golgi lumen attains a steady state.     

Phosphorylation of troponin and the effects of interactions between the components of the complex

1. The troponin complex from skeletal muscle contains approximately 1 mol of phosphate/80000g of complex, covalently bound to the troponin T component. 2. On prolonged incubation of the troponin complex or troponin T with phosphorylase kinase the phosphate content of troponin T was increased to approx. 3mol/mol. 3. On prolonged incubation of troponin I with phosphorylase kinase up to 1.6mol of phosphate/mol were incorporated. 4. Phosphorylation of troponin I was greatly inhibited by troponin C owing to the strong interaction between these proteins. Thus in the troponin complex troponin T was the main substrate for phosphorylase kinase. The phosphorylation of isolated troponin T was also inhibited by troponin C. 5. Troponin I was phosphorylated when the troponin complex was incubated with a bovine cardiac 3′:5′-cyclic AMP-dependent protein kinase. Troponin T either in its isolated form or in the troponin complex was not phosphorylated by bovine protein kinase to any significant extent under the conditions used. 6. If the troponin complex was dephosphorylated to 0.2mol/mol, or phosphorylated up to 2.5mol/mol there was no significant effect on the ability of normal concentrations to confer Ca²⁺ sensitivity on the adenosine triphosphatase of densensitized actomyosin.     

Analysis of the association of syncollin with the membrane of the pancreatic zymogen granule

Syncollin is a pancreatic zymogen granule protein that was isolated through its ability to bind to syntaxin. Here we show that syncollin has a cleavable signal sequence and can be removed from granule membranes by washing with sodium carbonate. When membranes were subjected to Triton X-114 partitioning, syncollin was found predominantly in the aqueous phase, indicating that it is not sufficiently hydrophobic to be embedded in the membrane. Syncollin has intramolecular disulfide bonds and was accessible to water-soluble cross-linking and biotinylating reagents only when granules were lysed by sonication. These results indicate that syncollin is tightly bound to the luminal surface of the granule membrane. In situ, syncollin was resistant to proteases such as trypsin. When granule membranes were solubilized in ionic detergents such as deoxycholate, this trypsin resistance was maintained, and syncollin migrated on sucrose density gradients as a large (150 kDa) protein. In contrast, in non-ionic detergents such as Triton X-100, syncollin became partially sensitive to trypsin and behaved as a monomer. Syncollin in alkaline extracts of granule membranes was also monomeric. However, reduction of the pH regenerated the oligomeric form, which was insoluble. We conclude that syncollin exists as a homo-oligomer and that its ability to self-associate can be reversibly modulated via changes in pH. In light of our findings, we reassess the likely role of syncollin in the pancreatic acinar cell.     

An autoradiographic study of the distribution of fibers from the dorsal motor nucleus of the vagus to the digestive tube of the rat

At the present time, complete agreement on the origin and course of parasympathetic preganglionic fibers to the alimentary canal has not been reached. The purpose of this study was to trace vagal fibers to the abdominal cavity and to follow the distribution of these fibers to the digestive tube. The technique used was to label neurons in the dorsal motor nucleus of the vagus (DMX) with 3H-leucine and then to follow the orthograde transport. 16 albino rats were used in this experiment. The right DMX in one group of rats and the left DMX in the other group was injected with 25 microCi of 3H-leucine in three injections. The injection sites and tissue sections from various areas of the digestive tube were processed for autoradiography. A heavy label was observed in the injection site and it could be traced down the vagus nerve through the thorax into the abdomen. Labelled vagal fibers were found in the parasympathetic ganglia of the stomach, small intestine and colon.     

Interaction of cells of the lymphoid organs in the production of antibodies against the influenza virus. Report 1. Localization of the virus in the cells of the spleen]

A study was made of localization of crude and inactivated influenza virus injected intravenously and intraperitoneally. Macrophages were found to engulf the virus penetrating into the spleen; this was shown by the quantitative and qualitative immunofluorescent method, elcetron microscopy and inoculation to syngenous recipients of homogenates of macrophagal and lymphocytic fractions of splenic cells. No virus was revealed in the lymphocytes.     

Effect of the Cerebral Tryptaminergic System on the Turnover of Dopamine in the Striatum of the Rat

The effect of the cerebral 5-hydroxytryptamine system on the turnover of striatal 3,4-dihydroxyphenyl-ethylamine (dopamine) was investigated by measuring the level of dopamine and one of its metabolites in rats depleted of cerebral 5-hydroxytryptamine or treated with a 5-hydroxytryptamine receptor blocker. Treatment with p-chlorophenylalanine induced, in addition to a reduction in striatal 5-hydroxytryptamine and 5-hydroxyindol-3-ylacetic acid, an increase in the striatal concentration of dopamine, a diminution in the concentration of homovanillic acid in the same cerebral area, and a reduction in the rise of this acid after the administration of a butyrophenone derivative or tetrabenazine. Treatment with methysergide also reduced the increase of homovanillic acid induced by the butyrophenone. When probenecid was given to rats treated with p-chlorophenylalanine, homovanillic acid failed to accumulate, whereas the accumulation of 5-hydroxyindol-3-ylacetic acid was unaffected. The decay of dopamine after alpha-methyl-p-tyrosine administration was normal for the first 6 h but was later reduced in rats given p-chlorophenylalanine or methysergide. The results suggest that the lack of activation of 5-hydroxytryptamine receptors leads to a reduction in the turnover of dopamine in the nigrostriatal pathway.     

Effect of triftazin on the transmission of excitation in the brain of the rabbit under the prolonged administration of the preparation]

Experiments were conducted on awake rabbits to which trifluoperazine was given daily for 2 weeks in a dose of 1 mg/kg a day; a study was made of the reaction of the cortical neurones in response to the electrostimulation of its adjacent areas. In measuring the dispersions of the poststimular histograms it was found that under the effect of trifluoperazine there occurred a reduction of the value and the duration of the responses of the cortical neurons after the infliction of stimulation. The effect obtained after the administration of trifluoperazine was regarded from the point of view of aggravation of the signal conduction in the neuronal reticula.     

Studies on the effect of manual massage of the ovaries on the reproductive activity of the mare

The aim of this study was to determine the influence of hand massage of mares' ovaries on breeding activity and hormonal changes in the winter anestrous period and during the luteal phase of the oestrus cycle. The experiment was conducted on 5 experimental and 5 control mares. In winter, (January) the experimental mares underwent 30-sec daily massage of both ovaries, for 30 d, and in summer (August) from the 6th day of the cycle to the occurrence of estrus. The sexual behavior of all mares was determined each day by individual teasing by a vigorous stallion, and the ovaries were checked by palpation per rectum and with an USG. Every second day blood samples were drawn from each mare to determine progesterone and estradiol in the plasma. Ovarian massage during deep winter anestrus had no significant effect on acceleration of the mares' active breeding season. Nevertheless, a higher concentration of estradiol was observed in the experimental group. These differences occurring on the 11th, 17th and 20th days were found to be significant (P ≤ 0.05). It was shown that during the summer period, in the luteal phase of the cycle, ovarian massage shortened the length of the estrous cycle, and ovulation was brought on somewhat earlier.     

Examination of the Possibility of the Involvement of the Alternative Respiration in Seed Germination

The possibility that the alternative respiration (AR) is involvedin seed germination was followed during and after imbibitionby noting the changes in the sensitivity to the respiratoryinhibitors, KCN and salicylhydroxamic acid (SHAM). This wascarried out by measuring oxygen uptake of mitochondrial preparationsfrom storage cotyledons and of the whole axis in a selectionof legumes (Vigna mungo, V. radiata, V. angularis, V. sinensisPhaseolus vulgaris, Pisum sativum and Dolichos lablab). Theextent of participation of AR in the respiration of cotyledonarymitochondria was variable according to species and time afterimbibition, and varied even within the same genus ( Vigna).In some species, AR was not observed. As to the axis, SHAM treatmentby itself had little effect on the rate of respiration, but,when applied with KCN, reduced oxygen uptake to an extent greaterthan KCN treatment alone. It was suggested that AR was not necessarilyimportant for seed germination. Key words: Alternative respiration, mitochondria, seed germination     

The effect of the nature of the strain on the character of the production of iron-dependent proteins by meningococci]

The comparative study of three Neisseria meningitidis strains (15, 125, 2394) was carried out by the method of electrophoresis in polyacrylamide gel in the presence of sodium dodecyl sulfate and by the method of immunoblotting. The intensive expression of 8 iron-regulated proteins (IRP) was shown to occur in iron-deficient culture medium. The major IRP with a molecular weight of 35 kD was expressed by all above-mentioned N. meningitidis strains under the conditions of iron deficiency and cross-reacted with 10 mouse and rabbit antisera to N. meningitidis of different groups, i.e. it was common to all Neisseria species. The antigenic activity of various IRP essentially differed with respect to antisera of animals and sera of patients with meningococcal infection.     

The importance of glucose in the oxidative metabolism of the testis of the conscious ram and the role of the pentose cycle

1. [U-(14)C]Glucose was infused into one or both testicular arteries of ten conscious rams and the specific activity of the glucose taken up by the testis was compared with the specific activity of the carbon dioxide produced by the testis. 2. Equilibration had occurred after infusion for 3hr. when a mean of 68% of the carbon dioxide was being derived by the testis from blood glucose and 86% of the glucose taken up by the testis was being oxidized to carbon dioxide. After 5hr. infusion, these values were 71% and 83% respectively. 3. In four other conscious rams, [1-(14)C]glucose was infused into one testicular artery and [6-(14)C] glucose into the other and the ;specific yields' of carbon dioxide calculated for the two forms of glucose. 4. From these values, it was calculated that a mean of 9.3% of the glucose taken up by the testis was metabolized via the pentose cycle.     

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.     

The structure of the cervical canal of the ewe

The cervical canal of the ewe does not allow for the consistent transcervical passage of insemination instruments. To define the factors affecting transcervical passage, the gross anatomy of the cervix and canal were studied in 100 estrous ewes and then in their reproductive organs following slaughter. In each ewe, the vagina and cervical opening was examined and the external os was classified into one of four types. Insemination instruments were introduced into the cervical opening and manipulation through the canal was attempted. Fluoroscopy was used to record the flow of contrast material through the canal. Ultrasound, xeroradiography and computed axial tomography were used to image the canal of the recovered reproductive tracts. Following imaging, each cervical canal was filled with silicone to create a mold which was used to measure and describe the canal. The average length (+/-SD) of the cervical canal was 6.7 +/- 1.1 cm and contained 4.9 +/- 1.0 funnel-shaped rings (n = 79). Successful passage of insemination instruments was limited by failure to identify the cervical opening and the small openings in the rings, 2.7 +/- 1.1 mm on average (+/-SD) which were not concentrically aligned. The eccentric rings were most consistently the second or third rings from the external os. The design of effective instrumentation and technique for transcervical passage must take these factors into account.