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Biomedical subjects

G Braun

Publications and source records attributed to G Braun.

At least 37 records · Page 2Linked to original sources

Acetylcholinesterase activity in neurons of crayfish abdominal ganglia.

Acetylcholine is known to be a neurotransmitter in crustacean central nervous systems, but the numbers and distribution of cholinergic neurons in the segmental ganglia have not been described. To begin a census of cholinergic neurons in these ganglia, we used a histochemical assay for acetylcholinesterase to map neurons that contained this enzyme in the six abdominal ganglia of crayfish. In each abdominal ganglion, about 47 cell bodies were stained. The distributions of these stained cells in individual ganglia were similar, and the numbers were not significantly different. None of these stained cell bodies could be identified from their structures or locations as previously identified motor neurons or sensory neurons with central cell bodies. The process of one unpaired midline neuron that occurred only in the first three abdominal ganglia divided to send a pair of axons anteriorly into both halves of the connective. The central projections of afferent axons from many peripheral sensory neurons stained clearly as they entered each ganglion. Terminals of these axons were heavily stained in the horseshoe neuropil and the lateral neuropils. We labeled both gamma-aminobutyric acid (GABA) and acetylcholinesterase in individual ganglia. Only a few neurons in each ganglion were double-labeled. The unpaired midline neurons in the three anterior ganglia that stained for acetylcholinesterase did not show GABA-like immunoreactivity, but cells with similar shapes did label with the GABA antiserum. Acetylcholinesterase is not a definitive marker of cholinergic neurons, but its presence is often associated with the cholinergic phenotype. These stained cells should be considered as putative cholinergic neurons.

Abdomen↗

Molecular characterisation and localisation of an Onchocerca volvulus pi-class glutathione S-transferase.

Glutathione S-transferases (GSTs) constitute a major detoxification mechanism in helminth organisms and are regarded vaccine candidates against helminth infections. Onchocerca volvulus glutathione-binding proteins were purified from the aqueous soluble fraction of homogenised adult females by affinity chromatography on glutathione-agarose. The eluted proteins had a specific GST activity of 1.6 mumol min-1 mg-1. Immunohistochemical studies localised these antigens in the hypodermis, the wall of the seminal receptacle and spermatozoa of adult worms. A lambda gt11 clone was isolated from an expression library of O. volvulus by immunoscreening. Sequence analysis revealed that it encoded a pi-class GST with 60% identity with Caenorhabditis elegans and up to 45% identity with mammalian pi-class GSTs. Antibodies affinity selected with recombinant GST demonstrated cross-reactivity between Litomosoides sigmodontis and O. volvulus GSTs.

Amino Acid Sequence↗

Food-induced arousal and nonassociative learning in honeybees: dependence of sensitization on the application site and duration of food stimulation.

Stimulus-induced arousal (sensitization) of a component of appetitive behavior in honeybees, the proboscis extension reflex (PER), was used to investigate different aspects of nonassociative memory. The sensitizing stimulus (sucrose solution) was applied to one antenna, as a compound to antenna and proboscis, and to the proboscis. Stimulus duration was either 1 or 3 s. Sensitization was evaluated by monitoring PER toward an odor before (pretest) and after (test) application of the sensitizing stimulus. All responses were quantified by recording from muscle M17 which represents the motor program of PER. Data were analyzed by determining (1) the response probability to the odor and (2) the response strength by determining the number of M17-spikes and the percentage of licking bees per trial. The analysis of the response probability led to two main results: the proportion of animals responding to the test odor depended on stimulus site, and, dependent on stimulus site, a longer application of the sensitizing stimulus resulted in different sensitization rates. The strength of the sensitized response, however, did not correspond to the probability, with which it was elicited, but rather to the strength of the response to the sensitizing stimulus itself. Furthermore, the three groups were not equally affected by the short and long stimulation. The analysis of the proportion of animals licking during test confirmed the data obtained using the number of muscle spikes as a measure of response strength. These results suggest an internal evaluation of the sensitizing stimulus depending on its quality and intensity. The differential effects after antennal and proboscis stimulation may be realized via an arousal system which has two independent functions, a permissive one modulating response probability and one modulating response strength. The permissive function of arousal may be regulated via an intervening inhibitory system whose activation critically depends on the functional significance of the arousing stimulus. The content of this short-term form of memory may be interpreted as an expectation for food which is regulated according to experienced consequences.

Animals↗

Effects of hydroxyapatite implants on primary mineralization during rat tibial healing: biochemical and morphometric analyses.

The effect of 40- to 60-mesh hydroxyapatite (HA) granules (Calcitek, Inc., Carlsbad, CA) on the process of primary mineralization during bone healing was examined following insertion of the HA granules into rat tibial bone after marrow ablation. Response to HA was assessed by monitoring morphometric and biochemical changes in matrix vesicles, which are extracellular organelles associated with initial calcification. Following insertion of HA, matrix vesicle-enriched membranes (MVEMs) were isolated from the tissue adjacent to the implant and from the endosteum of the contralateral limb at 3, 6, 14, and 21 days and from a nonimplanted control group (t = 0). MVEM alkaline phosphatase- and phospholipase A2-specific activities were increased on days 6 (peak) and 14; phosphatidylserine content was also elevated on days 6 and 14 (peak). Comparable changes were seen in the contralateral limb but at lesser magnitudes. Morphological changes were observed as well. The number of matrix vesicles/micron2 matrix increased on days 6 (peak) and 14. The mean diameter of the matrix vesicles was elevated on days 6 (peak), 14, and 21. Mean distance from the calcifying front increased on day 6 but was decreased on days 14 and 21. These results indicated that HA behaves like bone-bonding implants in that there is a stimulation of matrix vesicle enzymes, increased phosphatidylserine content, and increase numbers of matrix vesicles. However, the increases occur only after 6 days postimplantation, indicating a delay in response when compared to normal healing. This delay is confirmed by the morphometric measurements. HA causes a reduction in the response associated with marrow ablation. In addition, the effects of HA are comparable locally and systemically but with different intensity. These observations suggest that osteogenic cells are able to compensate for the inhibitory effects of HA and primary calcification involves normal matrix vesicle production and maturation, if somewhat delayed and reduced in magnitude. The ability to support primary mineral formation may contribute to the successful bonding of HA with surrounding osseous tissue.

Alkaline Phosphatase↗

Cross-reactive antigens in the pathogenesis of onchocerciasis.

The ocular disease associated with infection with Onchocerca volvulus is unique in that there is a wealth of epidemiological evidence to support the casual nature of the association but there is little known about the pathogenic mechanisms involved. We have identified a 44,000 M(r) component of ocular tissues that shows immunological cross-reactivity with an O. volvulus antigen. This immunological cross-reactivity between parasite and a component of host tissues may underlie the development of ocular disease in onchocerciasis. Preliminary experiments indicate that it is possible to initiate ocular disease in susceptible rats using the recombinant parasite antigen. This should allow the development of a laboratory model of ocular onchocerciasis and further our understanding of the mechanisms by which an infective organism can produce an auto-immune-like disease in the host.

Animals↗

Cholinergic modulation of the swimmeret motor system in crayfish.

1. The muscarinic agonist pilocarpine induced the swimmeret motor pattern in resting isolated preparations of the crayfish abdominal nerve cord and modulated the burst frequency in a dose-dependent manner. 2. Nicotine did not elicit rhythmic activity in resting isolated preparations but increased the burst frequency in active preparations. Nicotine produced higher burst frequencies than pilocarpine. 3. The acetylcholine (ACh) analogue carbachol combined the effects of pilocarpine and nicotine. It activated isolated resting preparations and increased the burst frequency as effectively as nicotine. The ACh-esterase inhibitor eserine also increased the burst frequency in active preparations. 4. Neither muscarinic nor nicotinic antagonists disrupted the proctolin-induced motor pattern, suggesting that proctolin and cholinergic agonists affect two different pathways for the activation of the swimmeret system. 5. We conclude that cholinergic interneurons participate in initiation of the swimmeret motor pattern and can modulate its burst frequency.

Acetylcholine↗

Immunologic cross-reactivity in the pathogenesis of ocular onchocerciasis.

PURPOSE: Onchocerca volvulus, a filarial worm, is a major cause of infectious blindness and inflammatory eye disease. An autoimmune cause for ocular onchocerciasis has been suggested since the identification of a recombinant antigen of O. volvulus that shows immunologic cross-reactivity with a host ocular component of 44,000 M(r). The aim of this study was to establish the distribution of the cross-reactive antigens in both host tissues and the parasite, and to determine if significant autoantibody responses to the host antigen could be detected in infected persons. METHODS: The tissue and organ distribution of the 44,000 M(r) antigen was determined by immunocytochemistry and Western blot analysis. Human autoantibody responses to the ocular antigen were demonstrated by Western blot analysis using sera collected from persons with onchocerciasis, with and without posterior segment pathology, Bancroftian filariasis, and Europeans with no filarial infection. RESULTS: The tissue distribution of the 44,000 M(r) antigen correlates with the sites of pathology in onchocerciasis and antibody reactivity against this antigen could be detected in all persons with onchocerciasis and posterior segment pathology. The antigen is also recognized by sera from persons with Bancroftian filariasis, but not from normal persons. CONCLUSIONS: A role is proposed for immunologic cross-reactivity in the pathogenesis of onchocerciasis and it is suggested that intraocular presentation of the cross-reactive parasite antigen by microfilariae is essential for the development of disease.

Adult↗

[Carbohydrate and lipid metabolism following heart bypass operations. The effect of the intravenous hypocaloric administration of glucose versus glucose xylitol (1:1)].

The effect of glucose-xylitol infusion on carbohydrate and lipid metabolism was investigated in 18 metabolically normal men (mean age 56.1 [35-65] years) with coronary heart disease after they had undergone a coronary artery bypass operation. During the first postoperative hours, group I (n = 6) received glucose only (2 mg/kg.min), group II (n = 6) glucose+xylitol (1 mg/kg.min each), and group II a glucose-containing electrolyte solution (0.83 mg/kg.min glucose). Blood glucose and insulin concentrations during the infusion period were significantly (P < 0.05) lower in groups II and III than I (glucose after 6 h: group I 21.5 [15.3-26.8] mmol/l; group II 14.2 [11.2-18.1] mmol/l; group III 12.6 [6.8-16.0] mmol/l). The highest lactate concentrations were reached in group I, 6 hours after the operation. Palmitine and stearine, as well as oleic and linoleic acid concentrations were significantly lower 12 hours postoperatively in group I than groups II and III (P < 0.05). These data indicate that energy-ineffective high glucose concentrations were avoided and endogenous lactate production reduced by the postoperative infusion of glucose+xylitol. In addition, it achieved a higher supply of free fatty acids as energy source to the myocardium without reaching toxic concentrations in the postischaemic myocardium.

Adult↗

A new way to monitor by-pass restorations of electron transport in inhibited chloroplasts by cyclic electron flow cofactors--a study by modulated fluorimetry.

Inhibition of electron transport in broken chloroplasts by DBMIB, under light-limiting conditions, is shown to be bypassed by PMS in a manner similar to the known effects of the phenylenediamine derivatives TMPD and DAD. These bypasses were demonstrated and further studied by modulated fluorimetry, monitoring DBMIB inhibition by the shift of the steady-state fluorescence towards the Fm level and the release of inhibition by a reverse shift together with establishment of a quenching effect by background far-red light. Comparative studies were also made with electron transport blocked by DCMU or BNT. A weak bypass by TMPD and a weaker one by PMS of the block created by DCMU was observed by modulated fluorimetry. The block created by BNT is similarly shown to be bypassed by TMPD but hardly or not at all by PMS. Bypass effects persisted even in the presence of ascorbate. It appears that, following reduction of the different cofactors by ascorbate in the stroma side, illumination caused the accumulation of a pool of oxidized cofactor molecules in the lumen, which is able to mediate electron transport between reduced plastoquinone and plastocyanin or P-700. The existence and the size of this pool were found to depend largely on the internal pH at the lumen, presenting an artificial system in which electron flow is controlled by the lumenal pH. The bypassing electron transport in the presence of DBMIB presumably avoids the participation of the cytochrome b6f complex. During its occurrence, there is also a strong imbalance in the activities of the two photosystems for linear electron flow, in favor of PS II. These experiments may thus serve to establish an in vitro model system for a future investigation of effects related to changes in the imbalance between the two photosystems and its regulation. Furthermore, this experimental system may also be utilized to study the role of the internal lumenal pH in control of photosynthesis.

Ascorbic Acid↗

Habituation of an appetitive reflex in the honeybee.

1. The proboscis extension reflex is an appetitive component of the bee's feeding behavior that is elicited by touching one antenna with a droplet of sugar water. Repetitive stimulation leads to a decrement and finally to the disappearance of the response, which can be restored by stimulating the contralateral antenna. This behavioral plasticity conforms to essential parametric characteristics for habituation. 2. The response was quantified by recording extracellularly from a muscle involved in proboscis movement, by measuring the duration of the proboscis extension, or by determining the number of trials necessary to abolish any visible response. 3. Because habituation was restricted to the repetitively stimulated antenna and did not generalize to the contralateral hemisphere, the neural circuits mediating habituation may be confined to one hemisphere. 4. State dependence of habituation could be demonstrated by showing that hungry animals exhibited a smaller response decrement and required more trials until disappearance of the response compared with satiated animals. The initial response and the subsequent response decrement are separate components determined by satiation level and stimulus strength. 5. Depleting the nervous system of monoamines by the use of reserpine abolished the reflex in 30% of the animals and reduced responsiveness in the remainder. Injection of octopamine or its metabolic precursor tyramine restored the reflex in reserpinized unresponsive animals, and tyramine also enhanced the muscle-spike discharge of reserpinized, responsive animals. In undepleted animals, tyramine application also accelerated the rate of habituation of the reflex. We therefore propose that octopaminergic neurons participate in mediating food arousal and the state dependence of habituation and, in a separate process, influence the response decrement during habituation. 6. Application of an acetylcholine esterase (AChE) inhibitor and a cholinergic receptor blocker confirmed histochemical data that implicate cholinergic transmission in the reflex pathways. 7. The combined pharmacological dissection of the reflex and immunocytochemical investigations of its chemical architecture provide evidence that the proboscis extension reflex is mediated by nonaminergic and monoaminergic pathways operating in parallel.

Animals↗

[Drash's syndrome and its variants. A report of 3 cases].

Three cases of Drash syndrome (DS) are presented. Two of them were 2 years old, the third one a newborn. Two are autopsy cases, one of the patients is alive. All three of them had nephrotic syndrome and renal failure, with mesangial sclerosis. All three had ambiguous genitalia, abdominal testes with variable degrees of dysgenesis and bilateral gonadoblastoma. Two cases had 46XY karyotype, in one case it was not performed. Two patients had nephroblastoma, one of them after several months of nephropathy, in the other it was the cause of consultation. The patient without nephroblastoma died at the age of 26 days with renal failure. We think that the original triad of DS: nephroblastoma, pseudohermaphroditism and nephropathy should be more precisely described and expanded to include related alterations in each one of its 3 elements: nephroblastomatosis, hamartomas, probably mesoblastic nephroma and pyelocaliceal malformations in addition to Wilms' tumor, variable disorders of sexual differentiation, gonadal dysgenesis and neoplasia, in addition to pseudohermaphroditism and superficial cortical renal dystrophy and increased reniculi, in addition to mesangial sclerosis. The various clinical forms of the syndrome are combinations of the triad in which a component can be missing and/or other related malformations be present. Chromosome anomalies are possibly responsible for this phenomena. Splitting into separate syndromes should not be done until the chromosome alterations and the regions and genes involved are identified.

Child, Preschool↗

Analysis of antibody responses directed against two Onchocerca volvulus antigens defined by monoclonal antibodies.

Monoclonal antibodies, reactive with antigens solubilised from the body wall of intact female Onchocerca volvulus using 2% 2-beta-mercaptoethanol, have been characterised. Two IgG1 antibodies, Cam1 and Cam28, recognised antigens of apparent molecular weights of 18,000 and 28,000; and 120,000, respectively. The target antigens of Cam1 and Cam28 could be localised in the cuticle. Inhibition ELISAs showed that target epitopes of both monoclonal antibodies induce an antibody response in onchocerciasis patients. 153 sera from Sierra Leonean patients were tested for their individual antibody levels against antigen epitopes recognised by Cam1 and Cam28. Patients within the age of 5-8 years had the highest levels of antibodies against the Cam28-epitope, whereas patients above 60 years had almost none. Amicrofilaremic patients had higher anti-Cam28 antibody levels than microfilaremic patients and there was a significant difference between groups with no chronic skin disease and those with mild or severe signs. A high percentage of patients (80.4%) recognised the Cam1-epitope, highest antibody levels being found in patients within the age group of 15 to 45 years and in microfilaremic patients. However, levels of antibodies inhibiting monoclonal antibody Cam1 could not be correlated with presence or absence of skin disease.

Adolescent↗

[Chromosomal translocation (2;13) in disseminated rhabdomyosarcoma].

A 12-year-old male with a disseminated alveolar rhabdomyosarcoma is reported. The diagnosis was difficult because of the clinical manifestations and the histological patterns of the bone marrow and a chest wall tumor. Diagnosis was confirmed through the histologic picture of a gum biopsy and the karyotype of the tumoral cells of the bone marrow. Chromosome study revealed a hypotetraploid cell line with the translocation (2;13) characteristic of this type of neoplasias. The usefulness of chromosome studies in solid tumors of childhood is emphasized.

Biopsy↗

Immunological crossreactivity between a cloned antigen of Onchocerca volvulus and a component of the retinal pigment epithelium.

Onchocerciasis (river blindness) is a major blinding disease in Africa, Central America, and South America. Loss of vision can be due to corneal change, optic atrophy, or chorioretinal disease. It has been suggested that autoimmunological reactions resulting from crossreactivity between parasite antigens and components of eye tissues contribute to development of ocular pathology. Using sera collected from onchocerciasis patients as a screening reagent, a cDNA clone (Ov39) has been isolated from a lambda gt11 expression library of Onchocerca volvulus. This antigen exhibits immunological crossreactivity with a component of retinal pigment epithelium cells (RPE). Antiserum raised against this recombinant peptide immunoprecipitates a 22,000 Mr antigen of adult O. volvulus and recognizes a 44,000 Mr component of bovine RPE by Western blotting. A 44,000 Mr antigen of cultured human RPE metabolically labeled with 35S-methionine can be immunoprecipitated with the same antiserum. An antigen of the same size is recognized by a rabbit antiserum raised against whole O. volvulus extract. Immunocytochemical studies on cryostat sections of the bovine eye using the antirecombinant sera localizes this antigen to the RPE.

Amino Acid Sequence↗

Proton flow through the ATP synthase in chloroplasts regulates the distribution of light energy between PS I and PS II.

The involvement of ATP synthase in the imbalance between the photoactivities of PS I and PS II under light-limiting conditions, was examined in broken lettuce chloroplasts using modulated fluorimetry. The imbalance, in favor of PS II, was minimal and roughly constant between pH 6.5-7.3 (ratio of PS II/PS I activities about 1.1), and maximal at pH 8.5 (ratio of PS II/PS I activities about 1.4). This increase was strongly inhibited by a treatment of the chloroplasts with the CF0 ATP synthase inhibitor DCCD, but unaffected by the CF1 ATPase inhibitor, tentoxin. However, tentoxin plus ADP-P1 did inhibit the high pH-induced increased imbalance. These results, when considered with the previous results on the effect of high pH on proton flux through the ATP synthase, suggest that the rate of such proton flow controls the imbalance between the two photo-systems. It is possible that there is an in vivo fine-tuning regulating mechanism of the photosystems imbalance via the opening and closing of proton gradient dissipation through the ATP synthase. This mechanism may help alleviate photoinhibitory damage.

Adenosine Diphosphate↗

[Schmuth's group findings--a simple system for classifying dysgnathias].

The advantages and disadvantages of Schmuth's findings, which classify eugnathic as well as dysgnathic dentition are opposed to Angle's classification. Using Schmuth's findings is a way of classifying without ignoring the physiological variability in the area of the first molars, which can lead to false diagnosis. 386 models were examined at the beginning of the patients' treatment. The results are discussed.

Dental Occlusion↗

Partial characterization of 2-beta-mercaptoethanol-soluble surface-associated antigens of Onchocerca volvulus.

Antigens were extracted from the epicuticle/cuticle of intact female Onchocerca volvulus using 2% 2-beta-mercaptoethanol and 1% SDS. In Western blot analysis a human infection serum selected for its high antibody titre against whole worm homogenates did not recognize any component solubilized by 1% SDS. However, the same serum did bind at least 7 antigens among the material extracted with 2-beta-mercaptoethanol. These antigens have apparent molecular weights (Mr) of: 15,000, 18,000, 28,000, 78,000, 98,000, 120,000 and 200,000. In ELISA using this preparation as target antigen, 151 out of 153 human infection sera gave positive results. An Onchocerca-specific IgG1 monoclonal antibody, designated Cam1, recognized the 28,000 Mr antigen, which is the most prominent antigen detected by Western blot analysis using human infection sera. In ELISA, using material affinity-purified with Cam1 as target antigen, 149 out of 153 human infection sera gave a positive IgG response. From a cDNA library three expressing clones were isolated with a rabbit serum raised against 2-beta-mercaptoethanol solubilized material. One of these clones was recognized by the monoclonal antibody Cam1.

Animals↗

[Sensitivity and specificity of CEA, Ca 15-3 and MCA levels in visceral breast carcinoma metastasis].

The determination of mucin like carcinoma associated antigen (MCA) showed a sensitivity of 72% in visceral metastasis of breast cancer, of 25% in metastasis of stomach cancer and of 10.3% in metastasis of colorectal cancer. The sensitivity of CA 15-3 was 83% (n.s.) in metastasis of breast cancer, that of CEA was 29% (p less than 0.05). The sensitivity of isolated metastasis and isolated invasion of the lymph nodes was under 20% for MCA, CA 15-3 and CEA.

Antigens, Neoplasm↗