[Biomarker in pulmonary diseases]. / Biomarker bei infektiösen und nicht infektiösen Lungenerkrankungen ausser Malignome.

Biological markers in various compartments of the human body have demonstrated potential value in diagnosis, prediction, guidance of therapy as well as in monitoring the clinical course of diseases of the airways and the lung. But only certain surrogate parameters are from clinical value, such as procalcitonin in pneumonia and sepsis, alpha-1-antitrypsin to diagnose alpha-1-antitrypsin deficiency, D-Dimers to detect emboli, nitric oxide in exhaled air in asthma, or isolation of germs from sputum to guide antibiotic treatment. Quantification of numerous markers in exhaled breath condensate or the detection of compounds in exhaled air are more recent attempts to further elucidate those biomarkers for clinical use. In general, biomarkers have an important supportive value in addition to routine diagnostic methods. The article reviews recent data regarding the usefulness of markers in non-malignant pulmonary diseases.

[Pulmonary function tests in the family doctor's office]. / Lungenfunktionstest als Orientierung bei der pneumologischen Diagnostik. Wie viel Luft haben Sie denn?

The basic diagnostic work-up including history-taking, clinical findings, and simple functional tests alone, suffices to enable the family doctor to rapidly differentiate between an obstructive and a restrictive pulmonary disease. This in particular by determining the "Tiffenau index", the ratio of the forced expiratory volume FEV1 to the forced vital capacity FVC (FEV1/ FVC). The question as to whether an obstructive condition is due to COPD or bronchial asthma is decided on the basis of the bronchospasmolytic test to determine the reversibility of the airways obstruction.

Orientation of carotenoids in the outer membrane of Synechocystis PCC 6714 (cyanobacteria).

The orientation of outer membrane carotenoids from Synechocystis PCC 6714 and Synechococcus PCC 6307 was studied by linear dichroism spectrophotometry. Uniaxially oriented, tilted outer membrane films revealed a significant linear dichroism after rotating the polarization vector of the incident light beam, indicating a predominant orientation of the carotenoid transition moments perpendicular to the outer membrane plane. Values for the reduced dichroism at the absorbance maxima presented a linear correlation to a function of the tilt angle (sin2 alpha).

Cloning and characterization of the genes coding for two porins in the unicellular cyanobacterium Synechococcus PCC 6301.

The genes somB and somA (Synechococcus outer membrane), lying in tandem organization in the genome of Synechococcus PCC 6301, encode two porins in the outer membrane of this unicellular cyanobacterium. Northern blot and primer extension experiments revealed that somA and somB are not comprising an operon, as each gene encodes a transcript of 1.7 kb length and has a distinct transcriptional start site. The deduced SomA and SomB protein sequences include typical N-terminal signal peptides and reveal 60% homology (50% identical residues) to each other as well as significant homology to six protein sequences deduced from open reading frames sequenced in the genome of the unicellular cyanobacterium Synechocystis PCC 6803. Furthermore, SomA possesses an overall identity of 97% to the functionally not yet characterized outer-membrane protein SomA from the closely related cyanobacterial strain Synechococcus PCC 7942. Analyses performed on the sequences suggest that SomA and SomB form 14- or 16-stranded porin-like beta-barrels. Moreover, all sequences share an N-terminal motif with significant homology to 'S-layer homology' domains, which might form a periplasmic extension. SomA and SomB therefore may, in addition to their porin function, act as linkers connecting the outer membrane with the peptidoglycan layer.

Afferent subcortical connections into the motor cortical larynx area in the rhesus monkey.

In three rhesus monkeys (Macaca mulatta), the inferior motor cortex was explored by electrical stimulation for sites yielding vocal fold adduction. The retrograde tracer wheat germ-agglutinin-conjugated horseradish peroxidase was injected into the effective sites. Within the forebrain, retrogradely labeled cells were found in the claustrum, basal nucleus of Meynert, substantia innominata, extended amygdala, lateral and posterior hypothalamic area, field H of Forel, and a number of thalamic nuclei with the strongest labeling in the nuclei ventralis lateralis, ventralis posteromedialis, including its parvocellular part, medialis dorsalis and centrum medianum, and weaker labeling in the nuclei ventralis anterior, ventralis posterolateralis, intermediodorsalis, paracentralis, parafascicularis and pulvinaris anterior. In the midbrain, labeling was found in the deep mesencephalic nucleus, ventral tegmental area, and substantia nigra. In the lower brainstem, labeled cells were found in the pontine reticular formation, median and dorsal raphe nuclei, medial parabrachial nucleus, and locus coeruleus. The findings are discussed in terms of the possible role of these structures in voluntary vocal control.

Afferent cortical connections of the motor cortical larynx area in the rhesus monkey.

The present study describes the cortical input into the motor cortical larynx area. The retrograde tracer horseradish peroxidase-conjugated wheat germ agglutinin was injected into the electrophysiologically identified motor cortical larynx area in three rhesus monkeys (Macaca mulatta). Retrogradely labeled cells were found in the surrounding premotor cortex (areas 6V and 6D), primary motor cortex (area 4), primary somatosensory cortex (areas 3, 1 and 2), anterior and posterior secondary somatosensory cortex and the probable homologue of Broca's area (areas 44 and 45); furthermore, labeling was found in the supplementary motor area, anterior and posterior cingulate cortex (areas 24 and 23), prefrontal and orbital frontal cortex (areas 8A, 46V, 47/12L, 47/12O, 13), agranular, dysgranular and granular insula as well as in the cortex within the upper bank of the middle third of the superior temporal sulcus (area TPO). The majority of these regions are reciprocally connected with the motor cortical larynx area [Brain Res 949 (2000) 23]. The laryngeal motor cortical input is discussed in relation to the connections of other motor cortical areas and its role in vocal control.

The anterior cingulate cortex and the phonatory control in monkey and man.

Electrical stimulation of the anterior cingulate cortex yields vocalization in the monkey. The elicited vocalizations seem to represent primary stimulus responses. Monkeys are not able to perform a vocal conditioning task after ablation of the anterior cingulate cortex. However, they can carry out a lever-pressing conditioning task following destruction of this area. It is hypothesized that the anterior cingulate cortex exerts the volitional control of species-specific vocalizations in monkey. The non-verbal emotional vocal utterances are considered to be the human homologue of monkey's vocalizations. Therefore, bilateral lesion of the anterior cingulate cortex in man should hamper the volitional control of emotional vocal utterances in man as it does in monkeys. One personal observation is reported where after a bilateral infarction of the anterior cingulate cortex the patient's voice showed a permanent lack of emotional expression. The anterior cingulate cortex seems to play the decisive role in the volitional verbalization of emotions.

Neuronal activity in the periaqueductal gray and bordering structures during vocal communication in the squirrel monkey.

In seven freely moving squirrel monkeys (Saimiri sciureus), the neuronal activity in the periaqueductal gray (PAG) and bordering structures was registered during vocal communication, using a telemetric single-unit recording technique. In 9.3% of the PAG neurons, a vocalization-correlated activity was found. Four reaction types could be distinguished: a) neurons, showing an activity burst immediately before vocalization onset; b) neurons, firing during vocalization, and starting shortly before vocalization onset; c) neurons, firing exclusively during vocalization; d) neurons, firing in the interval between perceived vocalizations (i.e. vocalizations produced by group mates) and self-produced vocal response. All PAG neurons showed a marked vocalization-type specificity. None of the neurons reflected simple acoustic parameters, such as fundamental frequency or amplitude, in its discharge rate. None of the neurons reacted to vocalizations of other animals not responded to by the experimental animal. All four reaction types found in the PAG were also found in the reticular formation bordering the PAG, though in lower density.

The role of the periaqueductal grey in limbic and neocortical vocal fold control.

The effects of a pharmacological blockade of the periaqueductal grey of the midbrain (PAG) on the elicitability of vocal fold movements from the facial motor cortex on the one hand and the anterior cingulate cortex and dorsomedial hypothalamus on the other were studied in the squirrel monkey. PAG blockade abolished vocal fold activity induced by the cingulate cortex and hypothalamus, but not that induced by the neocortex. These results point to the existence of two separate vocal fold control pathways at midbrain level: one limbic, responsible for non-verbal emotional vocal utterances, and one neocortical, responsible for the production of learned vocal patterns. The PAG represents a crucial relay station of the limbic but not the neocortical vocal control pathway.

On the search for the vocal pattern generator. A single-unit recording study.

In the squirrel monkey (Saimiri sciureus), single-unit activity was compared between the midbrain periaqueductal grey and the parvocellular and central nuclei of the medullary reticular formation during the production of species-specific vocalization. It was found that all three areas contain neurones with vocalization-related activity. The relative number of specific reactions types differed between areas, however. While the majority of periaqueductal cells fired just before, but not during vocalization, most cells in the reticular formation fired before and during vocalization. Modulation of discharge rate with changing fundamental frequency was only found in the reticular formation, not the periaqueductal grey. It is concluded that the parvocellular and central nuclei of the reticular formation, but not the periaqueductal grey are involved in vocal pattern generation.

Cell wall constituents of Microcystis sp. PCC 7806.

Cell walls of Microcystis sp. PCC 7806 were purified from cell homogenates by sucrose density centrifugation and Triton X-100 extraction. The outer membrane contained carotenoids, two major peptidoglycan-associated proteins (Mr 49,000 and 52,000), and lipopolysaccharide (LPS) as indicated by the presence of 3-hydroxy fatty acids (3-OH-14:0, 3-OH-16:0, 3-OH-18:0), 4-oxo-18:0 fatty acid, and GlcN as lipid A components in addition to rare O-methyl sugars (2-O-methyl-6-deoxyhexoses I and II). The peptidoglycan (A1 gamma-type) was found to be covalently linked to a wall polysaccharide composed of GlcN, ManN, Man, Glc, and phosphate.

Localization and distribution of hopanoids in membrane systems of the cyanobacterium Synechocystis PCC 6714.

Intracellular localization of triterpenic membrane stabilizers of the hopane series is described for the first time for a cyanobacterium. In Synechocystis PCC 6714, a bacteriohopanetetrol derivative (main compound) and diplopterol were detected in cell wall (CW) and thylakoid membrane (TM). Both hopanoids were enriched 4.5-fold and 9.0-fold in CW and outer membrane (OM) fractions, respectively, compared to TMs.

Chemical brain stimulation as a means to circumvent electrical stimulation artefacts in single-unit recording studies of evoked vocalization.

This study describes a combined single-unit recording/chemical brain stimulation technique that proved to be helpful in cases in which the behavior pattern to be studied electrophysiologically is difficult to elicit repetitively by external stimuli, but is easily obtained by brain stimulation. The advantage of this technique over electrical elicitation of the behavior is the avoidance of periodic stimulation artefacts that make detailed analyses of pattern-correlated neuronal activity often impracticable. Out of a number of substances tested for chemical brain stimulation, kainic acid proved to be the most effective one. With a single injection of 50 ng (in 200 nl water) into the periaqueductal gray of the squirrel monkey up to 6480 vocalizations could be obtained over a period of 65 min. Up to 22 injections could be made at this dose before a site became unresponsive.

Dual-channel telemetry system for recording vocalization-correlated neuronal activity in freely moving squirrel monkeys.

A miniature telemetric system is described which allows simultaneous measurements of neural activity and vocalization in freely moving monkeys within their social group. Single and multi-unit activities were detected with medium impedance electrodes that were fixed to self-made microdrives allowing accurate vertical positioning over a range of 8 mm. Vocalizations were registered by means of a piezo-ceramic device sensing the vocalization-induced skull vibrations. This allowed identification of the vocalizing animal in a larger group and eliminated environmental noise. Neuronal activity and vocalization were transmitted via separate channels of a FM transmitter using different carrier frequencies. The signals were decoded in two conventional FM receivers equipped with an automatic frequency control. The signals were stored for off-line analysis on a HiFi videotape recorder.

The role of the periaqueductal grey in vocal behaviour.

This is a review of our current knowledge about the role of the periaqueductal grey (PAG) in vocal control. It shows that electrical stimulation of the PAG can evoke species-specific calls with short latency and low habituation in many mammals. The vocalization-eliciting region contains neurones the activity of which is correlated with the activity of specific laryngeal muscles. Lesioning studies show that destruction of the PAG and laterally bordering tegmentum can cause mutism without akinesia. Neuroanatomical studies reveal that the PAG lacks direct connections with the majority of phonatory motoneurone pools but is connected with the periambigual reticular formation, an area which does have direct connections with all phonatory motor nuclei. The PAG receives a glutamatergic input from several sensory areas, such as the superior and inferior colliculi, solitary tract nucleus and spinal trigeminal nucleus. Glutamatergic input, in addition, reaches it from numerous limbic structures the stimulation of which also produces vocalization, such as the anterior cingulate cortex, septum, amygdala, hypothalamus and midline thalamus. Pharmacological blocking of this glutamatergic input causes mutism. The glutamatceptive vocalization-controlling neurones are under a tonic inhibitory control from GABAergic neurones. Removal of this inhibitory input lowers the threshold for the elicitation of vocalization by external stimuli. A modulatory control on vocalization threshold is also exerted by glycinergic, opioidergic, cholinergic, histaminergic and, possibly, noradrenergic and dopaminergic afferents. It is proposed that the PAG serves as a link between sensory and motivation-controlling structures on the one hand and the periambigual reticular formation coordinating the activity of the different phonatory muscles on the other.

Individual hemispheric asymmetry in vocal fold control of the squirrel monkey.

In ten squirrel monkeys (Saimiri sciureus), electrodes were implanted into the left and right facial motor cortex at sites producing bilateral vocal fold adduction when electrically stimulated. All animals, in addition, had electrodes in the periaqueductal grey of the midbrain (PAG) at sites producing vocalization when electrically stimulated. In eight out of ten animals, motorcortical stimulation during periaqueductally elicited vocalization caused a change in vocal output. This change, in four cases, was more severe with left-sided ipsilateral motor cortex/PAG stimulation than with right-sided ipsilateral stimulation. In the other four cases, right-sided stimulation was more effective than left-sided stimulation. It is concluded that the majority of squirrel monkeys show a hemispheric asymmetry in vocal fold control with left- and right-sided dominance distributed about equally in the population.

Neuronal activity in the medulla oblongata during vocalization. A single-unit recording study in the squirrel monkey.

In six squirrel monkeys (Saimiri sciureus), the medulla oblongata was explored with microelectrodes, looking for vocalization-correlated activity. The vocalizations were elicited by microinjections of glutamate agonists into the periaqueductal grey of the midbrain. Vocalization-related cells were found in greater numbers in the nucl. ambiguus (Ab) and retroambiguus (RAb), in the parvocellular, magnocellular and central reticular formation as well as in the solitary tract nucleus and spinal trigeminal nucleus. Small numbers were also found in the vestibular complex, cuneate nuclei, inferior olive and lateral reticular nucleus. A differentiation of the neuronal responses into 12 reaction types reveals that the frequency of each reaction type varies from brain structure to brain structure, thus allowing a specification of the different vocalization-related areas. According to this specification, it is proposed that initiation of vocalization takes place via the parvocellular reticular formation; vocal pattern control is mainly brought about by the parvocellular reticular formation, Ab, solitary tract nucleus and spinal trigeminal nucleus; expiratory control and respiratory-laryngeal coordination is carried out by the RAb, Ab and central nucleus of the reticular formation; vocalization-specific postural adjustments are carried out via the vestibular and cuneate nuclei.

Amygdalar vocalization pathways in the squirrel monkey.

In 22 squirrel monkeys (Saimiri sciureus) vocalization-eliciting electrodes were implanted into the amygdala and along the trajectory of the stria terminalis. Then, lesions were placed in the stria terminalis, its bed nucleus, the ventral amygdalofugal pathway and several di- and mesencephalic structures in order to find out the pathways along which the amygdala exerts its vocalization-controlling influence. It was found that different call types are controlled by different pathways. Purring and chattering calls, which express a self-confident, challenging attitude and an attempt to recruit fellow-combatants in intra-specific mobbing, respectively, are controlled via the stria terminalis; alarm peep and groaning calls, in contrast, which indicate flight motivation and resentment, respectively, are triggered via the ventral amygdalofugal fibre bundle. Both pathways traverse the dorsolateral and dorsomedial hypothalamus, respectively, and unite in the periaqueductal grey of the midbrain.

The efferent and afferent connections of the supplementary motor area.

The efferent and afferent connections of the supplementary motor area (SMA) were studied in 6 squirrel monkeys using [3H]leucine and horseradish peroxidase, respectively. Efferent projections, common to all leucine-injected animals, were found to the cortical areas 9,8,44,4,2,5,7,24 and 23. Subcortically , efferents were found to the putamen, caudate nucleus, claustrum, the thalamic nuclei reticularis, ventrialis anterior, ventralis lateralis, medialis dorsalis, centralis lateralis, paracentralis , centrum medianum, parafascicularis, centralis superior lateralis, centralis inferior and lateralis posterior, the subthalamic nucleus, field H of Forel, nuel . ruber, reticular formation of midbrain, pons and medulla, the pontine gray and nucl . reticularis tegmenti pontis. Afferent connections exist with the cortical areas 9,8,6,44,4,1,2,5,7, 24 and 23, insula, fronto-parietal operculum and superior temporal sulcus. Subcortical afferent connections exist with the claustrum, nucleus of the diagonal band, nucl . basalis Meynert, basolateral amygdaloid nucleus, the thalamic nuclei ventralis anterior, ventralis lateralis, medialis dorsalis, centralis lateralis, paracentralis , centrum medianum, centralis superior lateralis, centralis inferior, lateralis posterior and pulvinaris , the posterior hypothalamus, ventral tegmental area, nucl . ruber pars parvicellularis , reticular formation of midbrain and pons, locus coeruleus and nucl . centralis superior Bechterew. The projections are discussed with respect to the possible role SMA plays in the voluntary initiation of motor actions.

Afferents to the cortical larynx area in the monkey.

In 3 squirrel monkeys (Saimiri sciureus) horseradish peroxidase was injected into the cortical larynx area within the lower sensorimotor face cortex. Retrogradely labeled cells were found in a continuous band extending all along the upper bank of the Sylvian fissure from Broca's area rostrally to the parietal association cortex (area 7) caudally. In addition, labeled cells were found in the ventrolateral prefrontal cortex, orbital cortex, anterior cingulate gyrus, supplementary motor area, insula and inferior temporal gyrus. Subcortically, labeled neurons were situated in the substantia innominata, basolateral amygdaloid nucleus, lateral and posterior hypothalamus, the thalamic nuclei ventralis lateralis, ventralis posteromedialis, medialis dorsalis, centralis lateralis, centralis inferior, parafascicularis and pulvinaris, the periventricular gray, reticular formation, nucl. annularis, nucl. centralis superior (Bechterew) and locus coeruleus. Many of these structures are connected with the cortical larynx area reciprocally. The possible phonatory role of some of them is discussed.