ABSTRACT
BACKGROUND AND PURPOSE: Neuroinflammation has been proposed as part of the pathogenesis of post-concussion symptoms (PCS), but the inflammatory response of the human brain to mild traumatic brain injury (mTBI) remains unknown. We hypothesized that a neuroinflammatory response is present in mTBI at 1-2 weeks post-injury and persists in patients with PCS. METHODS: We scanned 14 patients with mTBI without signs of structural damage at 1-2 weeks and 3-4 months post-injury and 22 healthy controls once using the single photon emission computed tomography tracer 123 I-CLINDE, which visualizes translocator protein (TSPO), a protein upregulated in active immune cells. PCS was defined as three or more persisting symptoms from the Rivermead Post Concussion Symptoms Questionnaire at 3 months post-injury. RESULTS: Across brain regions, patients had significantly higher 123 I-CLINDE binding to TSPO than healthy controls, both at 1-2 weeks after the injury in all patients (P = 0.011) and at 3-4 months in the seven patients with PCS (P = 0.006) and in the six patients with good recovery (P = 0.018). When the nine brain regions were tested separately and results were corrected for multiple comparisons, no individual region differed significantly, but all estimated parameters indicated increased 123 I-CLINDE binding to TSPO, ranging from 2% to 19% in all patients at 1-2 weeks, 13% to 27% in patients with PCS at 3-4 months and -9% to 17% in patients with good recovery at 3-4 months. CONCLUSIONS: Neuroinflammation was present in mTBI at 1-2 weeks post-injury and persisted at 3-4 months post-injury with a tendency to be most pronounced in patients with PCS.
Subject(s)
Brain Concussion/diagnostic imaging , Brain/diagnostic imaging , Inflammation/diagnostic imaging , Adult , Aged , Brain/metabolism , Brain Concussion/metabolism , Bridged Bicyclo Compounds, Heterocyclic , Female , Humans , Inflammation/metabolism , Male , Middle Aged , Molecular Imaging , Post-Concussion Syndrome , Tomography, Emission-Computed, Single-Photon , Young AdultABSTRACT
Rapid HIV testing may circumvent the practical barriers to HIV testing in several settings. User preference of the testing kits available has been relatively underexplored. We examined healthcare provider (HCP) ratings of two validated rapid testing kits in clinical practice. From 1 July to 1 December 2012 we prospectively recruited HCPs (clinic nurses) from three outpatient clinics linked to Lausanne University Hospital, Lausanne, Switzerland. The HCPs had experience in taking blood samples but varying experience in rapid HIV testing. Participating HCPs performed rapid HIV testing using Determine™ Combo (DETE) or INSTI™ (INSTI), according to a predefined randomization sequence, and rated practical aspects of each test using a Likert scale. Seventeen HCPs of 23 approached (74%) were eligible and agreed to participate, performing a total of 336 HIV tests. Globally, the testing procedure was rated as easy or very easy by 97% (DETE) to 99% (INSTI) of tests performed. Among experienced HCPs, DETE was rated easier than INSTI for kit storage (p < 0.001) and blood collection ( P = 0.012) while INSTI was rated easier than DETE for blood application ( P = 0.001) and test interpretation ( P = 0.005). Among less experienced HCPs, both tests performed equally with the exception of test interpretation ( P < 0.001) and overall ease of use ( P = 0.05) in favour of INSTI. Of all HCPs, 94% stated they would recommend INSTI over DETE based on the time to result, ease of test interpretation and overall ease of use. Rapid HIV testing was considered easy to perform, even by inexperienced nursing staff. Whilst both tests were considered easy to use, the HCPs in this study preferred INSTI to DETE overall, due to rapid time to result, ease of test interpretation and general ease of use.
Subject(s)
AIDS Serodiagnosis/standards , Diagnostic Tests, Routine/methods , HIV Antibodies/blood , HIV Infections/diagnosis , HIV-1/immunology , Mass Screening/methods , Point-of-Care Systems , Reagent Kits, Diagnostic , HIV Infections/immunology , HIV Infections/virology , Health Personnel , Humans , Serologic TestsABSTRACT
Rhodococcus equi is a facultative intracellular bacterium causing severe pyogranulomatous pneumonia, ulcerative enterocolitis, and mesenteric lymphadenopathy in foals aged less than 6 months. Less frequently, this pathogen affects various other species, such as pigs, cattle, cats, and even humans. Although rhodococcosis is treated with a combination of antimicrobial agents, resistance is developed in some cases, and thus, antimicrobial susceptibility must be monitored and managed. Considering these limitations of the current therapy and unavailability of a vaccine to prevent the disease, research is particularly focused on the development of an effective vaccine against rhodococcosis. Most vaccines undergoing development utilize the virulence-associated protein (Vap) A antigen, which was identified previously as a key virulence factor of R. equi. Nevertheless, other proteins, such as VapG, present in most virulent R. equi strains, are also encoded by vap genes located on the R. equi bacterial virulence plasmid. In the present study, we evaluated the effect of VapG immunization on the survival of R. equi-challenged mice. We used attenuated Salmonella as a carrier for VapG (Salmonella-vapG+), a procedure previously adopted to develop a VapA-based vaccine. We observed that vaccination with Salmonella-vapG+ induced both an increased IFN-γ, IL-12, and TNF-α production, and a decreased bacterial burden in organs of the R. equi-challenged mice. Nevertheless, Salmonella-vapG+ vaccination protected only 50% of the mice challenged with a lethal dose of R. equi. Interestingly, we observed an increased frequency of B cells in the spleen of Salmonella-vapG+-vaccinated mice and showed that Salmonella-vapG+-vaccinated R. equi-challenged B-cell-knockout mice did not reduce the bacterial burden. Given these results, we discussed the potential role of the humoral immune response induced by Salmonella-vapG+ vaccination in conferring protection against R. equi infection, as well as the employment of VapG antigen for obtaining hyperimmune plasma to prevent rhodoccocosis in young foals.
ABSTRACT
AIM: The purpose of this study was to determine the effect of eccentric muscle damage on muscle activation patterns and intermuscular coherence during a fatiguing isometric contraction involving the elbow flexor muscles. METHODS: Ten young subjects participated in three experimental sessions that involved the performance of maximum voluntary contractions (MVCs), a constant-force task at 30% MVC, and a fatiguing isometric contraction at 30% MVC. The three sessions were performed before, 2 h after and 2 days after eccentric exercise to induce muscle damage in elbow flexor muscles. Task performance was quantified with electromyography (EMG) from the elbow flexor (biceps brachii, brachialis and brachioradialis) and extensor (triceps brachii) muscles, M-wave amplitude of biceps brachii, elbow flexor force fluctuations and endurance time of a fatiguing contraction. Intermuscular coherence during the fatiguing contraction was quantified from the rectified surface EMGs between muscle pairs. RESULTS: Eccentric exercise resulted in several indicators of muscle damage, such as a prolonged decline in muscle strength and an increase in muscle soreness 2 days after exercise. A 29% reduction in endurance time was observed 2 h after eccentric muscle damage, which returned to baseline 2 days later. The reduced endurance time 2 h after muscle damage was accompanied by an increase in EMG-EMG coherence between biceps brachii and brachialis muscles, which was observed at the end of the fatiguing contraction. CONCLUSION: These findings suggest that eccentric muscle damage produces a decrease in endurance time that is accompanied by an increase in intermuscular coherence in the presence of fatigue.
Subject(s)
Exercise/physiology , Isometric Contraction/physiology , Muscle Fatigue/physiology , Muscle, Skeletal/injuries , Muscle, Skeletal/physiology , Adolescent , Adult , Electromyography , Humans , Male , Young AdultSubject(s)
Lipoma/surgery , Median Nerve/surgery , Peripheral Nervous System Neoplasms/surgery , Diagnosis, Differential , Female , Humans , Image Processing, Computer-Assisted , Lipoma/diagnostic imaging , Median Nerve/diagnostic imaging , Middle Aged , Paresthesia/diagnostic imaging , Paresthesia/etiology , Paresthesia/surgery , Peripheral Nervous System Neoplasms/diagnostic imaging , UltrasonographyABSTRACT
An 85-year-old man with myasthenia gravis was successfully treated with methotrexate (10 mg/week), pyridostigmine and prednisolone (0-30 mg/day) for over 10 years. Then, he developed dysphagia and lost weight. Gastroscopy revealed Candida esophagitis. The patient received nystatin for 2 weeks. Methotrexate was stopped, and immunosuppressive therapy was continued with prednisolone alone. The patient has now remained in good condition for over 1 year. Although dysphagia is a typical symptom of myasthenia gravis, swallowing disturbances should not be attributed hastily to this disease, since they may also be a complication of therapy.
Subject(s)
Candidiasis/complications , Candidiasis/diagnosis , Deglutition Disorders/diagnosis , Deglutition Disorders/etiology , Esophagitis/complications , Esophagitis/diagnosis , Myasthenia Gravis/diagnosis , Aged , Antifungal Agents/therapeutic use , Candidiasis/drug therapy , Diagnosis, Differential , Drug Therapy, Combination , Esophagitis/drug therapy , Gastroscopy , Humans , Immunosuppressive Agents/therapeutic use , Male , Methotrexate/therapeutic use , Myasthenia Gravis/drug therapy , Nystatin/therapeutic use , Prednisolone/therapeutic use , Pyridostigmine Bromide/therapeutic useABSTRACT
An integral component of the manufacture of a skin substitute is the cryopreservation of the complete skin construct. Under this demand, investigations were carried out in the present work in the case of cryopreservation of human fibroblasts and keratinocytes composed to organotypical skin substitutes (OTS). Two scaffolds made up of gelatine and collagen/elastin were seeded with human fibroblasts via centrifugation method. Subsequent human keratinocytes were applied on the preceded scaffolds and cultivated under air-exposed conditions. For the investigation of the cryopreservation, OTS were frozen after 10 days cultivation via computer-controlled CryoMed included defined freezing conditions. After 24 hours storage in fluid nitrogen the OTSs were thawed and recultivated under airlift conditions. After that metabolic activity and immunfluorescent staining was analyzed in comparison with conventionally produced OTSs on basis of collagengel and/or OTSs based on scaffolds without cryopreservation. It could be assessed that cryopreservation has no negative influence on vitality and differentiation capacity of the cultivated constructs. The determination of OTS vitality after 14 days airlift culture delivered persistent higher metabolic activities of the scaffold based constructs in comparison with the corresponding controls. This could be confirmed by investigation of OTSs with and without cryopreservation. All expression patterns of differentiation marker could be detected after cryopreservation and subsequent recultivation. The results from cryopreservation of OTSs introduced here prove the possibility of temporally independent tailor-made applications by means of a complete skin substitute for example in the area pharmascreening.
Subject(s)
Cryopreservation , Skin/cytology , Tissue Engineering , Tissue Scaffolds , Cells, Cultured , HumansABSTRACT
BACKGROUND: All organisms living under aerobic atmosphere have powerful mechanisms that confer their macromolecules protection against oxygen reactive species. Microorganisms have developed biomolecule-protecting systems in response to starvation and/or oxidative stress, such as DNA biocrystallization with Dps (DNA-binding protein from starved cells). Dps is a protein that is produced in large amounts when the bacterial cell faces harm, which results in DNA protection. In this work, we evaluated the glycosylation in the Dps extracted from Salmonella enterica serovar Typhimurium. This Dps was purified from the crude extract as an 18-kDa protein, by means of affinity chromatography on an immobilized jacalin column. RESULTS: The N-terminal sequencing of the jacalin-bound protein revealed 100% identity with the Dps of S. enterica serovar Typhimurium. Methyl-alpha-galactopyranoside inhibited the binding of Dps to jacalin in an enzyme-linked lectin assay, suggesting that the carbohydrate recognition domain (CRD) of jacalin is involved in the interaction with Dps. Furthermore, monosaccharide compositional analysis showed that Dps contained mannose, glucose, and an unknown sugar residue. Finally, jacalin-binding Dps was detected in larger amounts during the bacterial earlier growth periods, whereas high detection of total Dps was verified throughout the bacterial growth period. CONCLUSION: Taken together, these results indicate that Dps undergoes post-translational modifications in the pre- and early stationary phases of bacterial growth. There is also evidence that a small mannose-containing oligosaccharide is linked to this bacterial protein.
Subject(s)
Carotid Artery, Internal, Dissection/complications , Carotid Artery, Internal, Dissection/diagnosis , Cranial Nerve Diseases/diagnosis , Cranial Nerve Diseases/etiology , Image Enhancement , Image Processing, Computer-Assisted , Imaging, Three-Dimensional , Magnetic Resonance Angiography , Accessory Nerve Diseases/diagnosis , Accessory Nerve Diseases/etiology , Adult , Carotid Artery, Internal/pathology , Diagnosis, Differential , Dominance, Cerebral/physiology , Female , Glossopharyngeal Nerve Diseases/diagnosis , Glossopharyngeal Nerve Diseases/etiology , Humans , Hypoglossal Nerve Diseases/diagnosis , Hypoglossal Nerve Diseases/etiology , Rupture, Spontaneous , Syndrome , Vagus Nerve Diseases/diagnosis , Vagus Nerve Diseases/etiologyABSTRACT
AIMS: To isolate bacteria capable of cleaving aliphatic carbon-sulfur bonds as potential biological upgrading catalysts for the reduction of molecular weight and viscosity in heavy crude oil. METHODS AND RESULTS: Thirty-one bacterial strains isolated from enrichment cultures were able to biotransform model compounds representing the aliphatic sulfide bridges found in asphaltenes. Using gas chromatography and mass spectrometry, three types of attack were identified: alkyl chain degradation, allowing use as a carbon source; nonspecific sulfur oxidation; and sulfur-specific oxidation and carbon-sulfur bond cleavage, allowing use as a sulfur source. Di-n-octyl sulfide degradation produced octylthio- and octylsulfonyl-alkanoic acids, consistent with terminal oxidation followed by beta-oxidation reactions. Utilization of dibenzyl sulfide or 1,4-dithiane as a sulfur source was regulated by sulfate, indicating a sulfur-specific activity rather than nonspecific oxidation. Finally, several isolates were also able to use dibenzothiophene as a sulfur source, and this was the preferred organic sulfur substrate for one isolate. CONCLUSIONS: The use of commercially available alkyl sulfides in enrichment cultures gave isolates that followed a range of metabolic pathways, not just sulfur-specific attack. SIGNIFICANCE AND IMPACT OF THE STUDY: These results give new insight into biodegradation of organosulfur compounds from petroleum and for biotreatment of such compounds in chemical munitions.
Subject(s)
Bacteria/metabolism , Chemical Industry , Industrial Microbiology/methods , Petroleum , Sulfides/metabolism , Aerobiosis , Biodegradation, Environmental , Bioreactors , Carbon , Molecular Weight , Sulfur , ViscosityABSTRACT
The objective of this study was the determination of the thermal regulatory and the thermal breakdown thresholds for in-tube restrained B6C3F1 and NMRI mice exposed to radiofrequency electromagnetic fields at 905 MHz. Different levels of the whole-body averaged specific absorption rate (SAR = 0, 2, 5, 7.2, 10, 12.6 and 20 W kg(-1)) have been applied to the mice inside the 'Ferris Wheel' exposure setup at 22 +/- 2 degrees C and 30-70% humidity. The thermal responses were assessed by measurement of the rectal temperature prior, during and after the 2 h exposure session. For B6C3F1 mice, the thermal response was examined for three different weight groups (20 g, 24 g, 29 g), both genders and for pregnant mice. Additionally, NMRI mice with a weight of 36 g were investigated for an interstrain comparison. The thermal regulatory threshold of in-tube restrained mice was found at SAR levels between 2 W kg(-1) and 5 W kg(-1), whereas the breakdown of regulation was determined at 10.1 +/- 4.0 W kg(-1)(K = 2) for B6C3F1 mice and 7.7 +/- 1.6 W kg(-1)(K = 2) for NMRI mice. Based on a simplified power balance equation, the thresholds show a clear dependence upon the metabolic rate and weight. NMRI mice were more sensitive to thermal stress and respond at lower SAR values with regulation and breakdown. The presented data suggest that the thermal breakdown for in-tube restrained mice, whole-body exposed to radiofrequency fields, may occur at SAR levels of 6 W kg(-1)(K = 2) at laboratory conditions.
Subject(s)
Body Temperature/radiation effects , Radio Waves , Animals , Dose-Response Relationship, Radiation , Electromagnetic Fields , Female , Hot Temperature , Male , Mice , Radiation Dosage , Radiation, Nonionizing , Radiometry , Risk , Temperature , Thermometers , Time FactorsABSTRACT
AIM: This study aimed to investigate the effectiveness of a one-week triple therapy with esomeprazole, clarithromycin and metronidazole for eradication of Helicobacter pylori infection in the absence of antimicrobial resistance. METHODS: Patients testing positive for H. pylori susceptible to metronidazole and clarithromycin (E-test) were randomized to receive a one-week regimen with either esomeprazole 2 x 20 mg or omeprazole 2 x 20 mg in combination with clarithromycin 2 x 250 mg and metronidazole 2 x 400 mg. Follow-up endoscopy with histology and culture and/or rapid urease test was performed 4-8 weeks after the end of treatment. RESULTS: Eighty patients were randomized. Helicobacter pylori infection was cured in 38/39 patients of the esomeprazole group and 31/33 patients of the omeprazole group (per protocol 97.4% (95% confidence interval [CI], 86.2-99.9), 93.7% (95% CI, 79.2-99.2), P=0.59); intention-to-treat 90.4% (95% CI: 77.4-97.3), 81.6% (95% CI: 65.7-92.3), respectively. No major side effects occurred. Minor side effects occurred in eight (20%) and six (23%) patients during esomeprazole and omeprazole therapy, respectively. Post-treatment susceptibility testing revealed resistance to both metronidazole and clarithromycin in two of the three patients who failed. CONCLUSION: We conclude that esomeprazole, clarithromycin and metronidazole as one-week triple therapy is effective for eradication of H. pylori in the absence of antimicrobial resistance.
Subject(s)
Anti-Infective Agents/administration & dosage , Anti-Ulcer Agents/administration & dosage , Helicobacter Infections/drug therapy , Helicobacter pylori , Omeprazole/administration & dosage , Adult , Aged , Clarithromycin/administration & dosage , Drug Resistance , Drug Therapy, Combination , Esomeprazole , Female , Humans , Male , Metronidazole/administration & dosage , Microbial Sensitivity Tests , Middle Aged , Patient Compliance , Treatment OutcomeABSTRACT
Neuronal nicotinic acetylcholine receptors (nAChRs) play a significant role in sympathetic transmission in the superior cervical ganglia (SCG), with most of the signal carried by a nAChR containing an alpha3 subunit. Work has shown that transection of the postganglionic nerves (axotomy) of the SCG results in a decrease in mRNA transcripts for alpha3, alpha5, alpha7 and beta4 and in protein expression of alpha7 and beta4. To evaluate effects of axotomy on alpha3 protein in the SCG, quantitative immunoblotting was used to demonstrate a dramatic decrease (> 80%) in the levels of this subunit 4 days after axotomy. Similarly, immunocytochemistry showed a marked decline in the number and the intensity of stained neurons for the alpha3 subunit as well as tyrosine hydroxylase. Ganglia explanted into culture for 4 days also showed a substantial decrease in alpha3 subunit protein. This decrease was partially prevented by the addition of nerve growth factor (NGF) to the culture medium at the time of explantation. Additionally, this decrease was reversed by the addition of NGF to the culture medium following 4 days in culture in the absence of NGF. These findings suggest that the loss of alpha3 subunit contributes to the reported decrease in ganglionic synaptic transmission that follows axotomy, and that NGF plays an important role in regulating the expression of alpha3-containing nAChRs in the SCG.
Subject(s)
Axotomy , Nerve Growth Factor/physiology , Neurons/metabolism , Receptors, Nicotinic/metabolism , Superior Cervical Ganglion/metabolism , Animals , Blotting, Western , Culture Techniques , Male , Nerve Growth Factor/pharmacology , Rats , Rats, Sprague-Dawley , Receptors, Nicotinic/drug effects , Reference Values , Superior Cervical Ganglion/drug effects , Sympathetic Nervous System/cytology , Sympathetic Nervous System/metabolismABSTRACT
A synthetic peptide corresponding to the C-terminus of the alpha 3 subunit of the rat neuronal nicotinic acetylcholine receptor (nAChR) was used to generate a rabbit polyclonal alpha 3 antibody. The specificity of this antibody was characterized by immunoblotting, immunohistochemical and immunoprecipitation techniques. Using this antibody, the relative densities of the alpha 3 subunit were quantitatively determined in different brain regions and in superior cervical ganglion (SCG). Among these regions, SCG, interpeduncular nucleus (IPN) and pineal gland showed the highest levels of alpha 3 protein expression. Habenula and superior colliculi had intermediate levels of expression. Low levels were found in cerebral cortex, hippocampus and cerebellum. The ontogenic profile of the alpha 3 subunit in the SCG was also determined. The alpha 3 protein level is low at postnatal day (P 1), but increases rapidly during the first seven postnatal days. This level then plateaus and remains stable through postnatal day 35. These findings suggest that neuronal nAChRs containing the alpha 3 subunit participate in important roles in specific regions of the rat brain and the SCG.
Subject(s)
Brain/metabolism , Neurons/metabolism , Receptors, Nicotinic/biosynthesis , Superior Cervical Ganglion/metabolism , Aging/metabolism , Animals , Antibody Specificity/immunology , Brain/cytology , Cell Line , Female , Habenula/metabolism , Humans , Kidney/cytology , Kidney/metabolism , Mesencephalon/metabolism , Organ Specificity , Pineal Gland/metabolism , Rats , Rats, Sprague-Dawley , Receptors, Nicotinic/analysis , Receptors, Nicotinic/genetics , Receptors, Nicotinic/immunology , Receptors, Nicotinic/metabolism , Superior Cervical Ganglion/cytology , Superior Colliculi/metabolism , TransfectionABSTRACT
KCNQ1 encodes KCNQ1, which belongs to a family of voltage-dependent K(+) ion channel proteins. KCNQ1 associates with a regulatory subunit, KCNE1, to produce the cardiac repolarizing current, I(Ks). Loss-of-function mutations in the human KCNQ1 gene have been linked to Jervell and Lange-Nielsen Syndrome (JLNS), a disorder characterized by profound bilateral deafness and a cardiac phenotype. To generate a mouse model for JLNS, we created a line of transgenic mice that have a targeted disruption in the Kcnq1 gene. Behavioral analysis revealed that the Kcnq1(-/-) mice are deaf and exhibit a shaker/waltzer phenotype. Histological analysis of the inner ear structures of Kcnq1(-/-) mice revealed gross morphological anomalies because of the drastic reduction in the volume of endolymph. ECGs recorded from Kcnq1(-/-) mice demonstrated abnormal T- and P-wave morphologies and prolongation of the QT and JT intervals when measured in vivo, but not in isolated hearts. These changes are indicative of cardiac repolarization defects that appear to be induced by extracardiac signals. Together, these data suggest that Kcnq1(-/-) mice are a potentially valuable animal model of JLNS.
Subject(s)
Disease Models, Animal , Long QT Syndrome/genetics , Potassium Channels, Voltage-Gated , Potassium Channels/physiology , Action Potentials , Animals , Base Sequence , DNA Primers , Ear, Inner/metabolism , Ear, Inner/pathology , Electrocardiography , Homeostasis/genetics , KCNQ Potassium Channels , KCNQ1 Potassium Channel , Long QT Syndrome/physiopathology , Mice , Mice, Mutant Strains , Mutation , Phenotype , Potassium Channels/geneticsABSTRACT
In vitro autoradiography using 125I-alpha-bungarotoxin (alpha-BGTx) and anti-alpha7 immunohistochemistry were performed on the dorsal motor nucleus of the vagus (DMV) of sham and chronically vagotomized rats to determine whether the alpha7-nicotinic acetylcholine receptor (nAChR) is located postsynaptically on DMV neurons whose axons contribute to the vagus nerve. Intense bilateral 125I-alpha-BGTx binding and anti-alpha7 immunostaining were observed in coronal brain sections containing the DMV of sham-vagotomized animals. Unilateral cervical vagotomy resulted in ipsilateral losses of 125I-alpha-BGTx binding and anti-alpha7 immunostaining from the DMV. Simultaneous staining of rat brainstem sections with anti-alpha7 and anti-choline acetyltransferase (ChAT) antibodies (to identify cholinergic DMV neurons that project into the vagus nerve) revealed that every DMV neuron that was stained for ChAT showed alpha7-staining as well. In vagotomized animals, no ChAT-positive neurons expressing alpha7-nAChRs remained in the ipsilateral DMV. We conclude that the alpha7-nAChR subtype is located postsynaptically on DMV neurons. To test whether the alpha7-nAChR is similar to the alpha7-homomeric nAChR, experiments were performed in anesthetized rats, and compounds were microinjected into the DMV while monitoring intragastric pressure (IGP). alpha-BGTx and strychnine antagonized nicotine-induced increases in IGP; no antagonism was observed with methyllycaconitine, a compound known to block the homomeric alpha7-nAChR subtype. Recovery from alpha-BGTx-induced antagonism of the nicotine response was observed. We conclude that there is a nAChR containing the alpha7-subunit in the DMV that is different from the homomeric alpha7-nAChR subtype.
Subject(s)
Aconitine/analogs & derivatives , Motor Neurons/metabolism , Receptors, Nicotinic/metabolism , Vagus Nerve/metabolism , Aconitine/pharmacology , Animals , Autoradiography , Bungarotoxins/pharmacokinetics , Bungarotoxins/pharmacology , Fluorescein-5-isothiocyanate , Fluorescent Antibody Technique, Indirect , Male , Microinjections , Nicotinic Agonists/pharmacology , Nicotinic Antagonists/pharmacology , Rats , Rats, Sprague-Dawley , Receptors, Nicotinic/drug effects , Stomach/drug effects , Vagotomy , Vagus Nerve/cytology , alpha7 Nicotinic Acetylcholine ReceptorABSTRACT
Epinephrine is a potent neurotransmitter and hormone that can influence cardiac performance beginning shortly after the first myocardial contractions occur in developing vertebrate embryos. In the present study, we provide evidence that the heart itself may produce epinephrine during embryonic development. Using antibodies that selectively recognize the catecholamine biosynthetic enzymes, tyrosine hydroxylase, dopamine ss-hydroxylase, and phenylethanolamine N-methyltransferase, we used coimmunofluorescent staining techniques to identify cardiac cells that have the capability of producing catecholamines. Initially, cells expressing catecholamine biosynthetic enzymes were found interspersed throughout the myocardium, but by embryonic day 11.5 (E11.5), they became preferentially localized to the dorsal venous valve and atrioventricular canal regions. As development proceeded, catecholamine biosynthetic enzyme expression decreased in these regions but became quite strong along the crest of the interventricular septum by E16.5. This expression pattern was also transient, decreasing in the ventricular septum by E19.5. These data are consistent with a transient and progressive association of catecholamine-producing cells within regions of the heart that become the sinoatrial node, atrioventricular node, and bundle of His. This is the first evidence demonstrating that intrinsic cardiac adrenergic cells may be preferentially associated with early pacemaking and conduction tissue development.
Subject(s)
Epinephrine/biosynthesis , Fetal Heart/metabolism , Animals , Dopamine beta-Hydroxylase/metabolism , Embryo, Mammalian/enzymology , Embryo, Mammalian/innervation , Embryo, Mammalian/metabolism , Female , Fetal Heart/enzymology , Fetal Heart/innervation , Fluorescent Antibody Technique , Heart Conduction System/embryology , Heart Conduction System/metabolism , Heart Conduction System/physiology , Male , Phenylethanolamine N-Methyltransferase/metabolism , Pregnancy , Rats , Rats, Sprague-Dawley , Time Factors , Tyrosine 3-Monooxygenase/metabolismABSTRACT
Initial F420-dependent hydrogenation of 2,4,6-trinitrophenol (picric acid) generated the hydride sigma-complex of picrate and finally the dihydride complex. With 2,4-dinitrophenol the hydride sigma-complex of 2,4-dinitrophenol is generated. The hydride transferring enzyme system showed activity against several substituted 2,4-dinitrophenols but not with mononitrophenols. A Km-value of 0.06 mM of the hydride transfer for picrate as substrate was found. The pH optima of the NADPH-dependent F420 reductase and for the hydride transferase were 5.5 and 7.5, respectively. An enzymatic activity has been identified catalyzing the release of stoichometric amounts of 1 mol nitrite from 1 mol of the dihydride sigma-complex of picrate. This complex was synthesized by chemical reduction of picrate and characterized by 1H and 13C NMR spectroscopy. The hydride sigma-complex of 2,4-dinitrophenol has been identified as the denitration product. The nitrite-eliminating activity was enriched and clearly separated from the hydride transferring enzyme system by FPLC. 2,4-Dinitrophenol has been disproven as a metabolite of picrate (Ebert et al. 1999) and a convergent catabolic pathway for picrate and 2,4-dinitrophenol with the hydride sigma-complex of 2,4-dinitrophenol as the common intermediate has been demonstrated.
Subject(s)
2,4-Dinitrophenol/metabolism , Actinomycetales/metabolism , Picrates/metabolism , Biodegradation, Environmental , Chromatography, High Pressure Liquid , Hydrogen-Ion Concentration , Magnetic Resonance Spectroscopy , NADH, NADPH Oxidoreductases/metabolism , NADP/metabolism , Nitrites/metabolism , Spectrophotometry, UltravioletABSTRACT
OBJECTIVE: Follistatin (FS) is the specific binding protein of activin and expression of both factors is regulated by inflammatory agents. Therefore, FS concentrations were determined in cerebrospinal fluid (CSF) of patients with bacterial and viral meningitis or multiple sclerosis (MS), as well as in the CSF of patients without meningial inflammation or autoimmune diseases. Furthermore, a mouse pneumococcal meningitis model was used to localise the cellular sources of FS in brains of normal and meningitic mice. METHODS: FS concentrations in CSF were determined by ELISA; FS in mice was localised by in situ hybridisation and immunohistochemistry. RESULTS: FS concentrations were > or =0.4 microg/l in 22 of 66 CSF samples of meningitis patients versus 2 of 27 CSF samples from patients with multiple sclerosis (P<0.05) and 2 of 41 CSF specimen from patients without neuroinflammatory diseases (P<0.01). In the CSF of patients with meningitis, the concentration of FS was correlated with total protein (P<0.005) and lactate concentrations (P<0.05), but not with leukocyte counts, interval between onset of disease and CSF analysis, or clinical outcome. The CSF-to-serum ratios of FS and albumin also correlated significantly (P<0.0005). In some patients with meningitis the CSF-to-serum ratios suggested that the elevated FS in CSF did not originate from serum alone. FS was localised in mice brains to neurones of the hippocampus, dentate gyrus, neocortex, and to the choroid plexus. Analyses of brains and other organs from uninfected and infected animals sacrificed 6-36 h after infection did not reveal any obvious differences in the distribution and intensity of FS mRNA and protein expression. CONCLUSIONS: The concentration of FS in humans is elevated during meningitis. In some patients the increase is caused by a release of FS from brain into CSF. Data from the mouse meningitis model suggest that increased CSF concentrations of FS in meningitis appear not to be accompanied by an elevated number of cells containing FS mRNA or protein in the brain.