The effects of electrical stimulation of the distal end of the cut sinus and aortic nerves on peripheral arterial chemoreceptor activity in the cat

1. In anaesthetized cats, stimulation of efferent components of the carotid sinus or aortic nerves depressed chemoreceptor discharge from the relevant chemoreceptor afferents. The local application of 2% procaine hydrochloride to the sinus nerve trunk, peripheral to the site of the stimulating electrodes and proximal to that of the afferent nerve twig, abolished the depression of afferent chemoreceptor discharge caused by electrical stimulation; on washing the procaine away electrical stimulation once more induced depression of chemoreceptor discharge.2. The depressant effect of efferent stimulation on carotid chemoreceptor activity was still seen during complete carotid glomeral ischaemia. Atropine given by close intra-arterial injection to the carotid body did not affect the depressant influence of efferent sinus nerve stimulation on carotid body chemoreceptor discharge.3. Stimulation of the sinus nerve efferents usually increased carotid body blood flow. Close arterial injection of atropine abolished this effect.4. The responses of glomeral blood flow and carotid chemoreceptor activity to efferent stimulation of the cut sinus nerve were not temporally related. It seems improbable that the depressant effect of such stimulation on chemoreceptor discharge was due to alterations of glomeral blood flow.5. Stimulation of the peripheral end of the cervical vagus in atropinized cats reduced chemoreceptor activity recorded in the ipsilateral aortic nerve.     

A comparison of primary afferent and cortical neurone activity coding sinus hair movements in the cat

Summary Responses in the somatosensory cortical area S I to stimulation of facial sinus hairs were recorded in the anaesthetized cat and compared with activity in primary afferent fibres innervating vibrissae follicles. The specific cortical vibrissa area is somatotopically organized; 39% of the cortical units in that area responded to stimulation of only a single sinus hair but in some cases all maxillary vibrissae activated a single cortical neurone. The responses consisted of three major groups; either a phasic discharge in response to the movement part of a stimulus, or an additional tonic discharge related to the steady period of vibrissa deflection, or a tonic discharge. On the basis of a comparison of response and excitability characteristics of primary afferent and cortical neurones it is concluded that all four kinds of peripheral units innervating sinus hair follicles project to the somatosensory cortical area S I. It appears from these findings that some cortical neurones receive a specific input related to a particular component of the complex primary afferent response in fibres innervating sinus hair follicles. The results are discussed with respect to previous reports on the central representation of facial sinus hairs in different species.     

Efferent and afferent impulse activity recorded from few-fibre preparations of otherwise intact sinus and aortic nerves

1. In anaesthetized cats, the efferent discharge recorded from slips of otherwise intact sinus nerves was sparse in eupnoeic conditions but increased markedly during systemic hypoxia or asphyxia or following the injection of cyanide or acetaldehyde into the circulation of the ipsilateral carotid body.2. When the sinus nerve was cut distal to the efferent slip the responses to cyanide or acetaldehyde were abolished. The sparse ;resting' activity which remained was increased following the intravenous injection of adrenaline. Following distal section, the impulse traffic of the efferent slip did increase during systemic hypoxia but the response was much feebler than when the nerve was otherwise intact.3. The impulse activity of most efferent slips, peeled off from the otherwise intact sinus nerve, was abolished when the glossopharyngeal nerve was cut central to its junction with the same sinus nerve, indicating that the activity was probably recorded from genuine efferent units. The discharge of some ;efferent' preparations was still present, however, following such section and showed an increase to local injections of cyanide. This activity was probably recorded from looping or branching chemoreceptor afferents.4. The discharge of efferent slips of the cut aortic nerve was increased following the intravenous injection of adrenaline and during systemic hypoxia. These responses were not present when the vagus nerve was cut central to the nodose ganglion.5. In eupnoea, chemoreceptor afferent activity recorded from slips of the sinus or aortic nerves is much the same whether these nerves be otherwise intact or whether they be cut. During systemic hypoxia, chemoreceptor afferent discharge was less when it was recorded from the otherwise intact nerves than when these nerve trunks were cut.6. The cell bodies of sinus nerve efferent fibres are synaptically excited by chemoreceptor afferents coursing in the same nerve trunk. The increase of efferent impulse activity aroused by this means depresses chemoreceptor afferent discharge.     

Adrenergic and pressure-induced modulation of carotid sinus baroreceptors in rabbit

The sympathetic control of the carotid sinus baroreceptor activity was studied in rabbits. Stimulation of the cervical sympathetic trunk (10 Hz, 1 ms, 4–12 V) elicited an increase in discharge frequency of the non-medullated baroreceptor afferents but not of the medullated fibers. An isolated carotid sinus area preparation was used to examine the influence of noradrenaline perfusion on baroreceptor activity. Non-medullated baroreceptor afferents, but not the medullated afferents, responded to noradrenaline perfusion (10^-6 g/ml) with a significant increase in firing rate. Short-term resetting of the baroreceptors was also studied with the same preparation. After perfusing the sinus at a hypertensive level for 30 min the pressure-response curves of both the non-medullated and the medullated baroreceptor afferents shifted to the right with increased thresholds and decreased maximal discharge frequencies. The extent of the short-term resetting was greater in non-medullated afferents than in medullated ones. It is concluded that the carotid sinus baroreceptors with non-medullated afferents are under the sympathetic control. The physiological significance of this is discussed.     

Characterization of cases of Clostridium difficile infection (CDI) presenting at an emergency room: molecular and clinical features differentiate community-onset hospital-associated and community-associated CDI in a tertiary care hospital

Definition of community-onset, hospital-acquired Clostridium difficile infection (CO-HA-CDI) is difficult in patients presenting with diarrhea at hospitals or outpatient clinics, especially 4 to 12 weeks after the last discharge. We performed C. difficile stool culture for 272 diarrheic patients visiting the emergency room (ER) between January 2006 and June 2010. C. difficile was isolated from 36 cases (13.2%), and isolation rates increased year by year, from 10.1% in 2008 to 12.4% in 2009 and 16.7% in 2010. Among 32 toxin-positive isolates, 13 (40.6%) and 19 (59.4%) were associated with CO-HA-CDI and community-acquired CDI (CA-CDI), respectively, if cases with CDI diagnosed within 12 weeks after discharge were considered hospital associated. The majority (70%) of CO-HA-CDI cases occurred within 2 weeks after hospital discharge, although the interval from discharge to onset of symptoms was as long as 10 weeks. We found via tcdA and tcdB and repetitive sequence PCR analysis, that toxin A-positive/toxin B-positive isolates were the most prevalent in both CO-HA-CDI (53.8%) and CA-CDI (94.7%) cases. Toxin A-negative/toxin B-positive isolates were also still highly associated with HA-CDI cases but were also observed in CA-CDI cases. Younger age, fewer underlying diseases, lack of prior antibiotic use, and genetic diversity of isolates in repetitive sequence PCR were the main characteristics in CA-CDI cases visiting the ER.     

Production of staphylococcal alpha toxin. II. Glucose repression of toxin formation

The effect of glucose on alpha toxin production was studied in the Wood 46 strain of Staphylococcus aureus. Optimal toxin production occurred when 0.2% glucose was present in the medium. Omission of glucose gave lower yields of toxin, and concentrations of 0.5% and higher severely depressed toxin formation. Glucose affected the initiation of alpha toxin synthesis in growing cultures. As the glucose concentration increased, the time lag prior to the onset of toxin production also increased, and maximal rates of synthesis were not obtained until essentially all the glucose had been exhausted from the medium. The addition of glucose to toxin-producing cultures caused a temporary, almost complete repression of toxin formation which was not due to pH changes in the culture. The synthesis of most extracellular proteins was not inhibited during the period of repression. After recovery, toxin was produced at rates equal to those of untreated control cultures. The kinetics of toxin repression and the observation that the glucose analogues, 2-deoxy-d-glucose and alpha-methyl-glucoside, as well as other carbon sources, inhibit toxin production suggest that transient repression is responsible for the inhibition of toxin formation. No evidence for a regulatory role of adenosine 3', 5'-cyclc monophosphate in alpha toxin production was obtained.     

Biochemistry of Vibrio cholerae Virulence III. Nutritional Requirements for Toxin Production and the Effects of pH on Toxin Elaboration in Chemically Defined Media

The investigations described in this report concern the metabolic and physiologic parameters governing cholera enterotoxin production in chemically defined growth media. The results indicate that the minimal nutritional requirements for growth of pathogenic Vibrio cholerae are the same as those for toxin production, and that toxin production parallels growth of the organisms. Studies of the relationship between toxin accumulation and pH reveal that toxin biosynthesis can be separated from toxin release. Toxin is synthesized below pH 7.0, but release and accumulation of extracellular toxin occur only at neutral or alkaline pH values.     

Toxins, butyric acid, and other short-chain fatty acids are coordinately expressed and down-regulated by cysteine in Clostridium difficile

It was recently found that a mixture of nine amino acids down-regulate Clostridium difficile toxin production when added to peptone yeast extract (PY) cultures of strain VPI 10463 (S. Karlsson, L. G. Burman, and T. Akerlund, Microbiology 145:1683-1693, 1999). In the present study, seven of these amino acids were found to exhibit a moderate suppression of toxin production, whereas proline and particularly cysteine had the greatest impact, on both reference strains (n = 6) and clinical isolates (n = 28) of C. difficile (>99% suppression by cysteine in the highest toxin-producing strain). Also, cysteine derivatives such as acetylcysteine, glutathione, and cystine effectively down-regulated toxin expression. An impact of both cysteine and cystine but not of thioglycolate on toxin yield indicated that toxin expression was not regulated by the oxidation-reduction potential. Several metabolic pathways, including butyric acid and butanol production, were coinduced with the toxins in PY and down-regulated by cysteine. The enzyme 3-hydroxybutyryl coenzyme A dehydrogenase, a key enzyme in solventogenesis in Clostridium acetobutylicum, was among the most up-regulated proteins during high toxin production. The addition of butyric acid to various growth media induced toxin production, whereas the addition of butanol had the opposite effect. The results indicate a coupling between specific metabolic processes and toxin expression in C. difficile and that certain amino acids can alter these pathways coordinately. We speculate that down-regulation of toxin production by the administration of such amino acids to the colon may become a novel approach to prophylaxis and therapy for C. difficile-associated diarrhea.     

Production of Staphylococcal Alpha Toxin I. Relationship Between Cell Growth and Toxin Formation

Alpha toxin production and its relationship to cell growth were studied in the Wood 46 strain of Staphylococcus aureus. Toxin first appeared in the culture in the late logarithmic stage, but at least 80% was produced during the subsequent period of slower cell growth. The toxin concentration per unit of cell mass or viable count increased continually throughout the period of toxin production and and at its maximum represented 1.6 to 2.0% of the dry weight of the cells. The possibility that alpha toxin is released as a result of cell lysis was examined by using the appearance of cellular deoxyribonucleic acid in the medium as an indicator of lysis. The results showed that no appreciable amount of lysis occurred during toxin production; at a time when almost maximum amounts of toxin were present in the culture, less than% 4 of the cells had lysed. This finding, together with the observation that less than 0.25% of the total amount of toxin in the culture could be found intracellularly, indicates that alpha toxin is released from intact cells shortly after it is synthesized.     

Effect of heptakis (2,6-O-dimethyl) beta-cyclodextrin on the production of pertussis toxin by Bordetella pertussis.

The effect of heptakis (2,6-O-dimethyl) beta-cyclodextrin (Me beta CD) on the production of pertussis toxin was evaluated. The addition of Me beta CD to the medium stimulated cell growth and pertussis toxin production. Me beta CD enhanced pertussis toxin production 100 times more in synthetic media, such as Stainer-Scholte medium (D. W. Stainer and M. J. Scholte, J. Gen. Microbiol. 63:211-220), than in Me beta CD-free medium in 2-day shake cultures. Maximum production of pertussis toxin was estimated as 50 mg of protein per liter of culture broth both by in vitro and in vivo assays. Purified toxin was demonstrated to be biochemically and biologically identical to the toxin produced in Me beta CD-free static cultures.     

Relationship of bacteriophages to alpha toxin production in Clostridium novyi types A and B.

The relationship of specific bacteriophages to the production of the lethal alpha toxin in Clostridium novyi types A and B was investigated. When type A strain 5771 reverted to the phage-sensitive state, it ceased to produce alpha toxin but continued to produce the gamma and epsilon antigens. This "nontoxigenic" culture, therefore, more closely resembled C. botulinum types C and D than the other C. novyi types. Phage-sensitive type B strains also ceased to produce the alpha toxin but continued to produce the beta toxin, and therefore very colesly resembled C. novyi type D (C. haemolyticum). Alpha toxin was again produced when the phage-sensitive cultures were reinfected with the respective tox+ phages. Alpha toxin production could also be induced in the "nontoxigenic" phage-sensitive derivatives from type B strain 8024 by tox+ phages isolated from other strains of type B. tox- phages were also isolated, but they did not affect alpha toxin production. The tox+ phages also caused a marked change in the colonial morphology of type B strains. In this report we present evidence that alpha toxin production by C. novyi type A strain 5771 and type B strain 8024 depends upon the continued presence and participation of specific bacteriophages designated as NA1tox+ and NB1tox+, respectively.     

Nonpathogenic Escherichia coli can contribute to the production of Shiga toxin

The food-borne pathogen, Escherichia coli O157:H7, has been associated with gastrointestinal disease and the life-threatening sequela hemolytic uremic syndrome. The genes for the virulence factor, Shiga toxin 2 (Stx2), in E. coli O157:H7 are encoded on a temperate bacteriophage under the regulation of the late gene promoter. Induction of the phage lytic cycle is required for toxin synthesis and release. We investigated the hypothesis that nonpathogenic E. coli could amplify Stx2 production if infected with the toxin-encoding phage. Toxin-encoding phage were incubated with E. coli that were either susceptible or resistant to the phage. The addition of phage to phage-susceptible bacteria resulted in up to 40-fold more toxin than a pure culture of lysogens, whereas the addition of phage to phage-resistant bacteria resulted in significantly reduced levels of toxin. Intestinal E. coli isolates incubated with Shiga toxin-encoding phage produced variable amounts of toxin. Of 37 isolates, 3 produced significantly more toxin than was present in the inoculum, and 1 fecal isolate appeared to inactivate the toxin. Toxin production in the intestine was assessed in a murine model. Fecal toxin recovery was significantly reduced when phage-resistant E. coli was present. These results suggest that the susceptibility of the intestinal flora to the Shiga toxin phage could exert either a protective or an antagonistic influence on the severity of disease by pathogens with phage-encoded Shiga toxin. Toxin production by intestinal flora may represent a novel strategy of pathogenesis.     

The effect of antigen on the carotid sinus receftors with reflex production of antibodies pharmacodynamically analyzed

Summary An operation of isolation of the carotid sinus with the following introduction of the physiologic saline into the carotid sinus has almost no effect on the titer of antibodies in the blood.Preliminary treatment of receptors of carotid sinus by Novocain and by sodium fluoride does not change its sensitivity to the antigenic stimulation. Introduction of antigen on this background brings about production of antibodies, the titer of which reaches the same level as in antigenic stimulation without preliminary treatment by these substances.Treatment of receptors of the isolated carotid sinus by cocaine, Dicaine and Sovcaline causes disapperance of the excitability of receptors to the antigenic stimulation. Introduction of typhoid vaccine into the carotid sinus which had been treated by cocaine, Dicaine and Sovcaine does not induce antibody production. The latter supports the view of the reflex mechanism of production of these antibodies.Presented by Academician A. D. Speransky     

Production of exoenzyme S by clinical isolates of Pseudomonas aeruginosa.

Exoenzyme S differs from toxin A and diphtheria toxin in that it does not adenosine diphosphate (ADP)-ribosylate elongation factor-2, but rather catalyzes the transfer of the ADP-ribose moiety of nicotinamide adenine dinucleotide to a number of different proteins in extracts of eucaryotic cells. Polyoma-transformed BHK-21 cells were isolated which were resistant to diphtheria toxin and toxin A. Extracts from these cells are ADP-ribosylated by exoenzyme S but not toxin A or diphtheria toxin, providing an assay which distinguishes between S and A activities. A total of 124 clinical isolates of P. aeruginosa were analyzed for production of toxin A and exoenzyme S. Exoenzyme S production was detected in 38% of the strains, whereas 80% of the strains produced toxin A.     

Modulation of IL-1-induced collagenase production in articular chondrocytes by pertussis toxin

The effects of pertussis and cholera toxins on interleukin-1 (IL-1) stimulated collagenase production from rabbit articular chondrocytes were investigated. Cholera toxin (50–1000 ng/ml) had no significant effect on IL-1-stimulated collagenase production. Pertussis toxin gave a 50–100% inhibition of collagenase activity induced by submaximal IL-1 concentrations. However, pertussis toxin had little effect on collagenase activity induced by maximal concentrations of IL-1. Optimal inhibitory effects were observed with 5–10 ng/ml of toxin.     

The electrophysiological analysis of the action of antigens on angioceptors

Summary Staphylococcal antigen produces irritation of the carotid sinus receptors; this is transformed into excitation, which gives rise to fluctuating bioelectric potentials of monophasic and biphasic character. The discharge period is increased, while in the intervals between the discharges the initial fluctuation of the potentials becomes intensified. Pertussis antigen also irritates the carotid sinus receptors and thus produces in the carotid sinus nerve a flow of impulses changing the normal oscillogram of this nerve.E. coli has a weak ability to irritate the carotid sinus receptors and changes only insignificantly the fluctuation rate of the carotid sinus nerve bioelectric potentials. The variations in the action currents of the carotid sinus nerve, originating from the action of the antigens on the carotid sinus receptors, are independent of any mechanical irritation. Administration of the same amount of physiological saline has no effect on the oscillogram of the sinus nerve.(Presented by Academician A. D. Speranskii) Translated from Byulleten' éksperimental'noi biologii i meditsiny Vol. 49, No. 2, pp. 90–94, February, 1960.     

The physiologically relevant information regarding systemic blood pressure encoded in the carotid sinus baroreceptor discharge pattern.

1. The objective was to find out what kind of informatioon regarding systemic blood pressure is transduced by baroreceptors in vivo and how this information is coded in the receptor discharge. 2. Carotid sinus pressure, e.c.g., and receptor action potentials were recorded for fifty-two single fibre carotid sinus receptors found in twenty decerebrated unanaesthetized cats. 3. The inflation and gradual deflation of an intraaortic catheter tip balloon manipulated the blood pressure in the carotid sinus in a way as to define the full in vivo stimulus-response curve for each receptor. 4. Correlation coefficients were computed between stimulus and response variables for several points on the response curve of each receptor and for every possible combination of stimulus and response variables defined. 5. Stimulus variables were (a) systolic, (b) diastolic,, (c) mean, (d) pulse pressures and (e) peak positive dP/dt. Response variables were (a) average discharge rat, (b) peak instantaneous frequency, and (c) average burst frequency. 6. For every fibre in the sample only the correlations between systolic, diastolic and mean pressures vs. average discharge rate were consistently high and positive. All other correlations were numerically low and/or negative. 7. It was concluded that in vivo baroreceptors signal mainly pressure level (systolic, diastolic or mean) as opposed to pulse pressure or dP/dt, and that the average discharge rate is their best index of information content.     

Method of isolating the carotid sinus and further proof of the reflex production of antibodies

Summary The method we suggest allows the complete isolation of the carotid sinus without injury to the sinus nerve. The administration of radioactive vaccine containing 100–400 C per 1 ml of radioactive phosphorus into the sinus isoiated by the method described above and the subsequent determination of the radioactivity of the blood testified to the complete absence of resorption from the isolated sinus. Preliminary section of the sinus nerve eliminated the production of specific agglutinins during the administration of typhoid fever vaccine into the isolated carotid sinus. Preliminary inclusion of the sinus nerve in the ligated bundle sharply deereased the production of antibodies. The data obtained testify to the presence of a reflex mechanism in the production of antibodies.Presented by Acting Member of the Academy of Medical Sciences of the USSR V. N. Chernigovsky     

Live varicella vaccine polarizes the mucosal adjuvant action of cholera toxin or its B subunit on specific Th1-type helper T cells with a single nasal coadministration in mice

This study was undertaken to determine whether the specific Th1- or Th2-cell response to varicella-zoster virus was induced predominantly by a mucosal adjuvant, cholera toxin, in mice. A commercially available live varicella vaccine (Oka strain) and cholera toxin or its B subunit were administered simultaneously via the nasal route. Delayed-type hypersensitivity to the Oka vaccine was induced, but the systemic neutralizing antibody response was low. The delayed-type hypersensitivity evoked after a single administration was relatively higher than that on administration three times. When spleen cells from mice immunized once with the vaccine and cholera toxin or its B subunit were restimulated with the live vaccine in vitro, there was greater thymidine uptake and production of interleukin- 2 (IL-2) than controls, but only a low level of IL-4 production. The production of IL-2 induced by the B subunit of cholera toxin was less than that by cholera toxin and a mutant of Escherichia coli enterotoxin on co-immunization with the vaccine in mice. Cholera toxin and its B subunit have been reported to induce predominantly a specific Th2-type T-cell response to various antigens. However, the Oka vaccine is an antigen that polarizes the activation of specific Th1/Th2-type T cells by cholera toxin or its B subunit to the Th1-type side. Cholera toxin and its B subunit are thus useful mucosal adjuvants for inducing cellular immunity to the Oka vaccine similar to Escherichia coli enterotoxin.     

Botulinum toxin A for axillary hyperhidrosis (excessive sweating)

BACKGROUND: Treatment of primary focal hyperhidrosis is often unsatisfactory. Botulinum toxin A can stop excessive sweating by blocking the release of acetylcholine, which mediates sympathetic neurotransmission in the sweat glands. METHODS: We conducted a multicenter trial of botulinum toxin A in 145 patients with axillary hyperhidrosis. The patients had rates of sweat production greater than 50 mg per minute and had had primary axillary hyperhidrosis that was unresponsive to topical therapy with aluminum chloride for more than one year. In each patient, botulinum toxin A (200 U) was injected into one axilla, and placebo was injected into the other in a randomized, double-blind manner. (The units of the botulinum toxin A preparation used in this study are not identical to those of other preparations.) Two weeks later, after the treatments were revealed, the axilla that had received placebo was injected with 100 U of botulinum toxin A. Changes in the rates of sweat production were measured by gravimetry. RESULTS: At base line, the mean (+/-SD) rate of sweat production was 192+/-136 mg per minute. Two weeks after the first injections the mean rate of sweat production in the axilla that received botulinum toxin A was 24+/-27 mg per minute, as compared with 144+/-113 mg per minute in the axilla that received placebo (P< 0.001). Injection of 100 U into the axilla that had been treated with placebo reduced the mean rate of sweat production in that axilla to 32+/-39 mg per minute (P<0.001). Twenty-four weeks after the injection of 100 U, the rates of sweat production (in the 136 patients in whom the rates were measured at that time) were still lower than base-line values, at 67+/-66 mg per minute in the axilla that received 200 U and 65+/-64 mg per minute in the axilla that received placebo and 100 U of the toxin. Treatment was well tolerated; 98 percent of the patients said they would recommend this therapy to others. CONCLUSIONS: Intradermal injection of botulinum toxin A is an effective and safe therapy for severe axillary hyperhidrosis.