Biomedical subjects

K Wild

Publications and source records attributed to K Wild.

At least 19 recordsLinked to original sources

PubMed2003

[Tissue toxicity of local anesthetics. An in vivo trial].

AIM: Local anesthetics are often administered intra-articularly after knee surgery. The aim of this study was to determine differences in irritancy of local anesthetics in an in vivo test performed on the chorioallantoic membrane of hen's eggs (HET-CAM test) to evaluate the specific irritation score. METHOD: After incubation for 9 days, the chorioallantoic membrane was prepared and then exposed to different local anesthetics (Naropin, Scandicain,Carbostesin,Xyloneural). RESULTS: We found no irritating values for the tested substances. No vascular injuries of the chorioallantoic membrane could be detected. Therefore, the irritation score was measured according to the standard protocol with an irritation score=0. CONCLUSION: Our results show that local anesthetics often used in clinics do not provoke severe vascular injuries as a sign of tissue toxicity. Therefore, the customary concentrations can be recommended for clinical use.

Allantois↗

PubMed2001

Crystal structure of an early protein-RNA assembly complex of the signal recognition particle.

The signal recognition particle (SRP) is a universally conserved ribonucleoprotein complex that mediates the cotranslational targeting of secretory and membrane proteins to cellular membranes. A crucial early step in SRP assembly in archaea and eukarya is the binding of protein SRP19 to specific sites on SRP RNA. Here we report the 1.8 angstrom resolution crystal structure of human SRP19 in complex with its primary binding site on helix 6 of SRP RNA, which consists of a stem-loop structure closed by an unusual GGAG tetraloop. Protein-RNA interactions are mediated by the specific recognition of a widened major groove and the tetraloop without any direct protein-base contacts and include a complex network of highly ordered water molecules. A model of the assembly of the SRP core comprising SRP19, SRP54, and SRP RNA based on crystallographic and biochemical data is proposed.

Amino Acid Sequence↗

PubMed2001

[Autonomic dysfunction with nocturnal dyspnea (Gerhardt-syndrome) in a patient with multiple system atrophy].

FINDINGS: This case involves a 56-year-old man with a 3-year case history, starting with vertigo, cerebellar symptoms, followed by inspiratory stridor, especially during the night, and autonomic failure with recurrent syncopes. Magnetic resonance imaging confirmed the diagnosis of olivopontocerebellar atrophy and tilting table tests with plasma catecholamine measurement revealed an extensive drop of blood pressure with no adequate change in the heart rate or plasma catecholamine, leading to the diagnosis of multiple system atrophy (MSA). In addition, bronchoscopy revealed that the inspiratory stridor was caused by paradoxical movement of the vocal cords (sometimes known as Gerhardt's syndrome). TREATMENT AND COURSE: The patient needed a tracheotomy and treatment of the autonomic symptoms. He could be discharged in better psychic condition, without syncopes or respiratory problems.

Diagnosis, Differential↗

PubMed2001

Linking sight and sound: fMRI evidence of primary auditory cortex activation during visual word recognition.

We describe two studies that used repetition priming paradigms to investigate brain activity during the reading of single words. Functional magnetic resonance images were collected during a visual lexical decision task in which nonword stimuli were manipulated with regard to phonological properties and compared to genuine English words. We observed a region in left-hemisphere primary auditory cortex linked to a repetition priming effect. The priming effect activity was observed only for stimuli that sound like known words; moreover, this region was sensitive to strategic task differences. Thus, a brain region involved in the most basic aspects of auditory processing appears to be engaged in reading even when there is no environmental oral or auditory component.

Adolescent↗

PubMed2001

Characterization of a new brain-derived proteoglycan inhibiting retinal ganglion cell axon outgrowth.

A proteoglycan was identified and isolated from physiological saline extracts of chick embryo brains by using a new monoclonal antibody (hybridoma clone mab Te38). The purified proteoglycan displayed an apparent molecular mass of 2500-3500 kDa, which became reduced to 370 and 600 kDa after digestion with chondroitinase ABC or chondroitinase AC. After additional treatment with keratanase the 600-kDa band was no longer detectable in Western blots. The specific epitope recognized by mab Te38 is an O-linked carbohydrate associated with the core protein. Tenascin-C, an extracellular matrix protein known to associate with several proteoglycans, copurified with the mab Te38 proteoglycan on the immunoaffinity column. Mab Te38 binds to the surface of nonneuronal cells; in sections from the primary visual system, expression is restricted to cells in the optic fissure, the dorsal optic nerve, and the chiasm. No retinal cells were found to express the mab Te38 epitope. The isolated molecule inhibited axon outgrowth from retinal explants when offered bound to a substrate consisting of either matrigel or collagen, chondroitinase treatment did not alter the inhibitory properties. The distribution and in vitro function of the Te38 proteoglycan indicate that it may serve a role in guidance of retinal ganglion cell axons.

Animals↗

PubMed2001

[Improved ultrasound imaging of the pancreas with the transsplenic view and Tissue harmonic imaging].

UNLABELLED: The conventional ultrasound examination of the pancreas from ventral is - especially concerning the leftsided parts of the organ - limited by interposition of stomach, flexura duodenojejunalis or colon. The transsplenic view to the cauda pancreatis is less known, but allows quite often a good possibility to examine this part of the retroperitoneal organ, even when examination from ventral is impossible. AIM: Comparison of ventral and transsplenic ultrasound examination - with and without tissue harmonic imaging (THI) - in regard to the diagnostic accessability to the cauda pancreatis. METHOD: Ultrasound examination of 135 patients from both positions with and without THI and comparison of the accessability by experienced examiners. RESULTS: As examination from the ventral position allows a proper view to the cauda only in 58 %, the transsplenic position allows an adequate view in 98 %. The examiners regarded THI as a useful improvement in examining the pancreas in 85 %. CONCLUSION: The transsplenic view to the pancreas is less known than the examination from the classical ventral transducer position. It enables the examinator quite often to examine the pancreatic cauda even when examination from ventral is impossible. Therefore it should be enclosed in the routine procedure of abdominal ultrasound examination. THI is a further possibility to improve the ultrasound access to the pancreas, especially when conditions for a good view and proper examination are limited.

Adolescent↗

PubMed2000

Structure and assembly of the Alu domain of the mammalian signal recognition particle.

The Alu domain of the mammalian signal recognition particle (SRP) comprises the heterodimer of proteins SRP9 and SRP14 bound to the 5' and 3' terminal sequences of SRP RNA. It retards the ribosomal elongation of signal-peptide-containing proteins before their engagement with the translocation machinery in the endoplasmic reticulum. Here we report two crystal structures of the heterodimer SRP9/14 bound either to the 5' domain or to a construct containing both 5' and 3' domains. We present a model of the complete Alu domain that is consistent with extensive biochemical data. SRP9/14 binds strongly to the conserved core of the 5' domain, which forms a U-turn connecting two helical stacks. Reversible docking of the more weakly bound 3' domain might be functionally important in the mechanism of translational regulation. The Alu domain structure is probably conserved in other cytoplasmic ribonucleoprotein particles and retroposition intermediates containing SRP9/14-bound RNAs transcribed from Alu repeats or related elements in genomic DNA.

Alu Elements↗

PubMed2000

MRI-based stereolithographic models of the temporomandibular joint: technical innovation.

A new technique of manufacturing dual-colour stereolithographic models of hard and soft tissues of the temporomandibular joint (TMJ) is presented. Sagittal T1/PD weighted magnetic resonance (MR) images of joints with and without disc displacement were obtained in the closed and open mouth positions. Individual interactive contour identification of bony structures and the articular disc followed by binary interpolation provided the data for the generation of acrylic TMJ models. Three dimensional in vivo visualization of the articular disc in relation to bony structures in the closed and open mouth positions allows a new perception of normal and pathological TMJ anatomy.

Acrylic Resins↗

PubMed1999

pH gating of ROMK (K(ir)1.1) channels: control by an Arg-Lys-Arg triad disrupted in antenatal Bartter syndrome.

Inward-rectifier K(+) channels of the ROMK (K(ir)1.1) subtype are responsible for K(+) secretion and control of NaCl absorption in the kidney. A hallmark of these channels is their gating by intracellular pH in the neutral range. Here we show that a lysine residue close to TM1, identified previously as a structural element required for pH-induced gating, is protonated at neutral pH and that this protonation drives pH gating in ROMK and other K(ir) channels. Such anomalous titration of this lysine residue (Lys-80 in K(ir)1.1) is accomplished by the tertiary structure of the K(ir) protein: two arginines in the distant N and C termini of the same subunit (Arg-41 and Arg-311 in K(ir)1.1) are located in close spatial proximity to the lysine allowing for electrostatic interactions that shift its pK(a) into the neutral pH range. Structural disturbance of this triad as a result from a number of point mutations found in patients with antenatal Bartter syndrome shifts the pK(a) of the lysine residue off the neutral pH range and results in channels permanently inactivated under physiological conditions. Thus, the results provide molecular understanding for normal pH gating of K(ir) channels as well as for the channel defects found in patients with antenatal Bartter syndrome.

Arginine↗

PubMed1999

The 2 A structure of helix 6 of the human signal recognition particle RNA.

BACKGROUND: The mammalian signal recognition particle (SRP) is an essential cytoplasmic ribonucleoprotein complex involved in targeting signal-peptide-containing proteins to the endoplasmic reticulum. Assembly of the SRP requires protein SRP19 to bind first to helix 6 of the SRP RNA before the signal-peptide-recognizing protein, SRP54, can bind to helix 8 of the RNA. Helix 6 is closed by a GGAG tetraloop, which has been shown to form part of the SRP19-binding site. RESULTS: The high-resolution (2.0 A) structure of a fragment of human SRP RNA comprising 29 nucleotides of helix 6 has been determined using the multiple anomalous dispersion (MAD) method and bromine-labelled RNA. In the crystal the molecule forms 28-mer duplexes rather than the native monomeric hairpin structure, although two chemically equivalent 11 base pair stretches of the duplex represent the presumed native structure. The duplex has highly distorted A-RNA geometry caused by the occurrence of several non-Watson-Crick base pairs. These include a 5'-GGAG-3'/3'-GAGG-5' purine bulge (which replaces the tetraloop) and a 5'-AC-3'/3'-CA-5' tandem mismatch that, depending on the protonation state of the adenine bases, adopts a different conformation in the two native-like parts of the structure. The structure also shows the 2'3'-cyclic phosphate reaction product of the hammerhead ribozyme cleavage reaction. CONCLUSIONS: The 29-mer RNA is the first RNA structure of the human SRP and provides some insight into the binding mode of SRP19. The observed strong irregularities of the RNA helix make the major groove wide enough and flat enough to possibly accommodate an alpha helix of SRP19. The variety of non-canonical base pairs observed enlarges the limited repertoire of irregular RNA folds known to date and the observed conformation of the 2'3'-cyclic phosphate containing Ade29 is consistent with the current understanding of the hammerhead ribozyme reaction mechanism.

Base Sequence↗

PubMed1999

Protein binding of piritramide: influence of various protein concentrations and the postoperative acute phase response.

OBJECTIVE: Piritramide is a lipophilic opioid, which is widely used for postoperative analgesia and analgosedation in Europe. In this study we investigated the influence of various protein concentrations (total protein, alpha1-acid glycoprotein, albumin) and the postoperative acute phase response on the protein binding of piritramide. METHODS: The influence of various protein concentrations on the protein binding of piritramide was investigated by either diluting the serum samples of five volunteers with isotonic saline or by adding different amounts of alpha1-acid glycoprotein. Albumin binding was measured in a 5% human albumin solution. The impact of the postoperative acute phase response was investigated by obtaining daily serum samples from 18 surgical patients until the third postoperative day, and measuring piritramide protein binding, alpha1-acid glycoprotein, total protein and albumin. RESULTS: There was a significant relationship between piritramide protein binding and the concentrations of total protein and alpha1-acid glycoprotein. The binding to albumin was 88%. During the postoperative acute phase response, the protein binding of piritramide did not change. Serum concentrations of alpha1-acid glycoprotein increased, whereas total protein and albumin decreased. CONCLUSION: Although there were significant changes in the piritramide-binding proteins, alpha1-acid glycoprotein and albumin, during the postoperative acute phase response, the protein binding of piritramide did not change. Therefore, a change in protein binding, which might be one factor to be considered in determining piritramide dosage in the postoperative period, does not have to be taken into account.

Acute-Phase Reaction↗

PubMed1999

Selective visual attention in patients with frontal lobe lesions or Parkinson's disease.

Visual selective attention and response competition were tested in patients with frontal lobe lesions or with Parkinson's disease, and matched normal controls. The target stimuli were presented with flanking distractors that were either compatible, incompatible, or neutral to the target stimulus. The distance between the target and distractors was systematically varied. A control condition without distractors was also included. Subjects' response times to target stimuli and accuracy were measured. Both patient groups responded significantly slower and less accurately than their respective matched normal controls across all interference conditions and spatial distances. However, they did not show significantly greater interference or facilitation effects. Thus, the data suggest that the cognitive processes underlying selective attention are, in general, spared in patients with frontal lobe lesions or basal ganglia dysfunction.

Adult↗

PubMed1999

Expression of membrane-bound and cytosolic guanylyl cyclases in the rat inner ear.

Membrane-bound guanylyl cyclases (GCs) are peptide hormone receptors whereas the cytosolic isoforms are receptors for nitric oxide. In the inner ear, the membrane-bound GCs may be involved in the regulation of fluid homeostasis and the cytosolic forms possibly play a role in signal processing and regulation of local blood flow. In this comprehensive study, we examined, qualitatively and quantitatively, the transcription pattern of all known GC isoforms in the inner ear from rat by RT-PCR. The tissues used were endolymphatic sac, stria vascularis, organ of Corti, organ of Corti outer hair cells, cochlear nerve, Reissner's membrane, vestibular dark cells, and vestibular sensory cells. We show that multiple particulate (GC-A, GC-B, GC-D, GC-E, GC-F and GC-G) and several subunits of the heterodimeric cytosolic GCs (alpha1, alpha2, beta1 and beta2) are expressed, albeit at highly different levels. GC-C was not found. GC-A and the soluble subunits alpha1 and beta1 were transcribed ubiquitously. GC-B was present in all tissues except stria vascularis, which contained GC-A and traces of GC-E and GC-G. GC-B was by far the predominant membrane-bound isoform in the organ of Corti (86%), Reissner's membrane (75%) and the vestibulum (80%). Surprisingly, GC-E, a retinal isoform, was detected in significant amounts in the cochlear nerve (8%) and in the organ of Corti (4%). Although the cytosolic GC is a heterodimer composed of an alpha and a beta subunit, the mRNA transcription of these subunits was not stoichiometric. Particularly in the vestibulum, the transcription of the beta1 subunits was at least four-fold higher than of the alpha1 subunit. The data are compatible with earlier suggestions that membrane receptor GCs may be involved in the control of inner ear electrolyte and fluid composition whereas NO-stimulated GC isoforms mainly participate in the regulation of blood flow and supporting cell physiology.

Animals↗

PubMed1999

Expression pattern of adenylyl cyclase isoforms in the inner ear of the rat by RT-PCR and immunochemical localization of calcineurin in the organ of Corti.

Most studies concerning adenylyl cyclases in the inner ear were carried out before the advent of molecular biology. In a PCR approach using cDNAs of six inner ear tissues (stria vascularis, endolymphatic sac, organ of Corti, vestibulum, cochlear and vestibular nerve) we found tissue specific expression of adenylyl cyclase isoforms. Adenylyl cyclases types 2 and 4 are predominant in the fluid controlling tissues, i.e. in the stria vascularis and endolymphatic sac. In the organ of Corti and vestibulum the Ca2+-modulated isoforms types 1, 6 and 9 were expressed. The regulation of adenylyl cyclase 9, which is the major isoform expressed in the organ of Corti, proceeds via the Ca2+-activated protein phosphatase 2B (calcineurin, PPP3). PCR with specific primers for calcineurin demonstrated its abundant expression in the organ of Corti. Using a monoclonal antibody we localized calcineurin immunochemically to the cochlear nerve, the nerve fibers and the inner hair cells. In the cochlear and vestibular nerves a characteristic neuronal expression pattern of adenylyl cyclase isoforms was observed, i.e. adenylyl cyclases types 2, 3 and 8. The functional consequences of the adenylyl cyclase expression pattern in the inner ear are discussed in conjunction with its unique sensory performance.

Adenylyl Cyclases↗

PubMed1999

Dry soda lime markedly degrades sevoflurane during simulated inhalation induction.

We have investigated gas composition during simulated inhalation induction with sevoflurane to elucidate possible mechanisms of incidental prolonged induction times and airway irritation. Using a circle system, 8% sevoflurane in oxygen 6 litre min-1 was washed into an absorbing canister filled with fresh soda lime containing 2.9% KOH (Draegersorb, 'D') or no KOH (< 0.01%, Sofnolime, 'S'). The absorbent was dried by oxygen 20,000 litre before every second experiment. Maximum soda lime temperatures attained after 4-6 min were 107 degrees C using dry D and 62 degrees C (61 degrees C) with dry S. Temperature did not increase with fresh soda lime. With dry soda lime, sevoflurane was not detected at the T-piece for 3 min and reached 6-7% within 6-10 min. After 1 min, we detected methanol and compound A (CH2F-O-C(= CF2) (CF3)). Total amounts over 20 min were: methanol 1125 mg (D dry), 334 mg (S dry) and < 5 mg (fresh soda lime); compound A 148 mg (D dry), 13 mg (S dry) and 3-8 mg (fresh); and fluoride 8.5 mg (D dry), 3.3 mg (S dry) and 1 mg (fresh). Formaldehyde was detected only with dry lime (D > 2.5 mg, S > 0.6 mg). In summary, the use of moist soda lime is of crucial importance during inhalation induction. With dry soda lime, the patient may inhale potentially toxic degradation products in significant amounts. Sevoflurane degradation is aggravated by a high KOH content of the lime. The observed airway irritation may be caused by formic acid, which is generated in isomolar concentrations with methanol (Cannizzaro reaction). The amount of compound A found with dry KOH-containing lime is unlikely to be noxious.

Anesthesia, Closed-Circuit↗

PubMed1998

The effect of anti-diuretic hormone on the endolymphatic sac of the inner ear.

The anti-diuretic hormone vasopressin (AVP) regulates water excretion from the kidney by increasing the water permeability of the collecting duct. AVP binds to V2-receptors and induces the translocation of aquaporin-2 water channels (AQP-2) into the apical plasma membrane of principal cells. By this mechanism AVP controls water reabsorption in the kidney. The effects of AVP on the endolymphatic sac (ES) of the inner ear, which is thought to mediate reabsorption of endolymph, were investigated. Both the V2-receptor and the AQP-2 water channel were found to be expressed in the ES epithelium. In the ES AVP binds to receptors most probably of the V2-subtype. Application of AVP to organotypically cultured ES inhibits membrane turnover in ribosomal-rich cells of the ES epithelia, which is thought to mediate translocation of AQP-2 into the surface membrane. This suggests that AVP has contrasting effects in the inner ear and kidney, which may be physiologically useful for maintaining endolymphatic pressure during severe hypovolemia. Animal experiments show that AVP causes endolymphatic hydrops after systemic application to guinea-pigs, which suggests a causal role for the increased AVP levels found in humans suffering from Ménière's disease.

Animals↗

PubMed1998

Metabolic engineering as therapy for inborn errors of metabolism--development of mice with phenylalanine hydroxylase expression in muscle.

Treatment of many inherited liver enzyme deficiencies requires the removal of toxic intermediate metabolites from the blood of affected individuals. We propose that circulating toxins can be adequately cleared and disease phenotype influenced by enzyme expressed in tissues other than the liver. Phenylalanine hydroxylase (PAH) activity was constitutively expressed in skeletal and cardiac muscle of transgenic mice which carried the PAH cDNA under the transcriptional control of the mouse muscle creatine kinase promoter. Muscle PAH-expressing mice were bred to liver PAH-deficient, hyperphenylalaninemic mice to yield progeny that lack PAH activity in liver but express PAH in muscle. These mice exhibited hyperphenylalaninemia at baseline, but serum phenylalanine levels decreased significantly when the mice were supplemented with tetrahydrobiopterin (BH4), a required cofactor for PAH. This is the first demonstration that a liver-specific enzyme, when expressed in a heterologous tissue and supplied with necessary cofactors, can effectively clear toxic metabolites from the circulation of individuals with inherited enzyme deficiency. This result suggests that gene therapy targeted to heterologous tissues, such as muscle, will be effective in the treatment of selected inborn errors of metabolism.

Animals↗

PubMed1997

Structure, catalysis and supramolecular assembly of adenylate kinase from maize.

The crystal structure of adenylate kinase from maize ligated with an inhibitor has been determined by molecular replacement and refined to 3.5-A resolution. The enzyme keeps the ATP/ADP/AMP equilibrium in the cell. In the C4 plant maize, it has the special task to recycle the AMP produced in large amounts in primary CO2 assimilation. The established structure explains the side reaction with CMP. Moreover, it shows infinite rods that can be readily discerned in the crystal packing. In comparison with homologues, two structural differences that are crucial for this supramolecular assembly are evident. We propose that the rods represent a natural inactive storage form that assembles at night when maize stops CO2 assimilation and thus most of the AMP production in its C4 cycle. The enzyme is particularly abundant in mesophyll chloroplasts, where such an assembly would release appreciable amounts of water that can be used in other processes during the night.

Adenylate Kinase↗