Substantial understory contribution to the C sink of a European temperate mountain forest landscape.

CONTEXT: The contribution of forest understory to the temperate forest carbon sink is not well known, increasing the uncertainty in C cycling feedbacks on global climate as estimated by Earth System Models. OBJECTIVES: We aimed at quantifying the effect of woody and non-woody understory vegetation on net ecosystem production (NEP) for a forested area of 158 km2 in the European Alps. METHODS: We simulated C dynamics for the period 2000-2014, characterized by above-average temperatures, windstorms and a subsequent bark beetle outbreak for the area, using the regional ecosystem model LandscapeDNDC. RESULTS: In the entire study area, woody and non-woody understory vegetation caused between 16 and 37% higher regional NEP as compared to a bare soil scenario over the 15-year period. The mean annual contribution of the understory to NEP was in the same order of magnitude as the average annual European (EU-25) forest C sink. After wind and bark beetle disturbances, the understory effect was more pronounced, leading to an increase in NEP between 35 and 67% compared to simulations not taking into account these components. CONCLUSIONS: Our findings strongly support the importance of processes related to the understory in the context of the climate change mitigation potential of temperate forest ecosystems. The expected increases in stand replacing disturbances due to climate change call for a better representation of understory vegetation dynamics and its effect on the ecosystem C balance in regional assessments and Earth System Models.

Genetically driven brain serotonin deficiency facilitates panic-like escape behavior in mice.

Multiple lines of evidence implicate brain serotonin (5-hydroxytryptamine; 5-HT) system dysfunction in the pathophysiology of stressor-related and anxiety disorders. Here we investigate the influence of constitutively deficient 5-HT synthesis on stressor-related anxiety-like behaviors using Tryptophan hydroxylase 2 (Tph2) mutant mice. Functional assessment of c-Fos after associated foot shock, electrophysiological recordings of GABAergic synaptic transmission, differential expression of the Slc6a4 gene in serotonergic neurons were combined with locomotor and anxiety-like measurements in different contextual settings. Our findings indicate that constitutive Tph2 inactivation and consequential lack of 5-HT synthesis in Tph2 null mutant mice (Tph2-/-) results in increased freezing to associated foot shock and a differential c-Fos activity pattern in the basolateral complex of the amygdala. This is accompanied by altered GABAergic transmission as observed by recordings of inhibitory postsynaptic currents on principal neurons in the basolateral nucleus, which may explain increased fear associated with hyperlocomotion and escape-like responses in aversive inescapable contexts. In contrast, lifelong 5-HT deficiency as observed in Tph2 heterozygous mice (Tph+/-) is able to be compensated through reduced GABAergic transmission in the basolateral nucleus of the amygdala based on Slc6a4 mRNA upregulation in subdivisions of dorsal raphe neurons. This results in increased activity of the basolateral nucleus of the amygdala due to associated foot shock. In conclusion, our results reflect characteristic syndromal dimensions of panic disorder and agoraphobia. Thus, constitutive lack of 5-HT synthesis influence the risk for anxiety- and stressor-related disorders including panic disorder and comorbid agoraphobia through the absence of GABAergic-dependent compensatory mechanisms in the basolateral nucleus of the amygdala.

[Physical and technical elements of short-interval, color-filtered double strobe flash-stroboscopy]. / Technisch-physikalische Grundlagen der Doppelbelichtungsstroboskopie.

INTRODUCTION: Quantitative measurement of vocal fold movements can be done either with high-speed imaging or with short interval, color-filtered double strobe flash-stroboscopy. The physical and technical elements of this new technique are described. METHODS: Two special strobe units (KAY Elemetrics RLS 9100) are used in a master-slave configuration. In this way an adjustable interval of 0.1-2.0 ms between flashes is introduced. The strobe flashes are color filtered and are separated by a brief interval. By this means a double exposure is created in each video frame.Real-time visualization of opening and closing velocities over the entire length of the vocal fold from anterior to posterior is possible. Quantification is possible off-line after image calibration. CONCLUSION: Short-interval, color-filtered double-strobe flash stroboscopy allows quantitative measurement of the velocity of vocal fold movements during vibration at different pitches and sound pressure levels (SPL). Images gained with this new technique provide information about a dynamic property (velocity) of the vocal fold within a single image.Therefore, its use could be helpful from the aspect of clinical documentation.

TBX1 is responsible for cardiovascular defects in velo-cardio-facial/DiGeorge syndrome.

Velo-cardio-facial syndrome (VCFS)/DiGeorge syndrome (DGS) is a human disorder characterized by a number of phenotypic features including cardiovascular defects. Most VCFS/DGS patients are hemizygous for a 1.5-3.0 Mb region of 22q11. To investigate the etiology of this disorder, we used a cre-loxP strategy to generate mice that are hemizygous for a 1.5 Mb deletion corresponding to that on 22q11. These mice exhibit significant perinatal lethality and have conotruncal and parathyroid defects. The conotruncal defects can be partially rescued by a human BAC containing the TBX1 gene. Mice heterozygous for a null mutation in Tbx1 develop conotruncal defects. These results together with the expression patterns of Tbx1 suggest a major role for this gene in the molecular etiology of VCFS/DGS.

Recurrent laryngeal nerve transposition in guinea pigs.

Improved control of prosthetic voice aids for laryngectomees might be possible to obtain with residual laryngeal motor nerve signals. We were able to recover motor signals from the recurrent laryngeal nerve (RLN) by transposing it into the ipsilateral denervated sternohyoid muscle (SH) in 8 guinea pigs. Reinnervation was monitored by electromyographic recordings from surface and intramuscular needle electrodes in awake animals. Within 4 to 14 weeks after surgery, all animals demonstrated laryngeal-like motor activity in the reinnervated SH, including activity during respiration, sniffing, swallowing, and/or vocalizing. After 3 to 6 months, the animals were reanesthetized, and nerve stimulation and section experiments confirmed the RLN as the source of reinnervation in all cases. In several animals, activity of the RLN-innervated SH was demonstrated to be correlated with that of contralateral laryngeal muscles. Histochemical analysis of the SH indicated a unilateral transformation from mostly fatigable to mostly fatigue-resistant fiber types ipsilateral to the RLN transposition, a phenotype more typical of laryngeal muscles. Thus, RLN transposition at the time of laryngectomy may be a method for salvaging laryngeal control signals that could be used to control prosthetic voice devices.

Inductively coupled plasma mass spectrometry: a versatile tool for a variety of different tasks.

In a laboratory that is working in many different fields, a systematic approach is needed to decide efficiently how a given analytical task should be handled. Four typical examples are presented to show how ICP-MS may be used to solve client's problems. The examples are: the identification of lead projectiles, the determination of total and leachable fractions of impurities in a polymer (PVDF), used for manufacturing components of ultrapure water distribution systems, the authentication of antique silver alloys, and the determination of rare earth elements in geological materials. These examples demonstrate not only typical challenges for the instruments and the analysts handling them, but also ways to reach a satisfactory solution within a reasonable amount of time.

Motoneuron axon distribution in the cat stapedius muscle.

Stapedius-motoneuron cell bodies in the brainstem are spatially organized according to their acoustic response laterality, as demonstrated by intracellular labeling of physiologically identified motoneurons [Vacher et al., 1989. J. Comp. Neurol. 289, 401-415]. To determine whether a similar functional spatial segregation is present in the muscle, we traced physiologically identified, labeled axons into the stapedius muscle. Ten labeled axons were visible in the facial nerve and five could be traced to endplates within the muscle. These five axons had 39 observed branches (others may have been missed). This indicates an average innervation ratio (> or = 7.8) which is much higher than that obtained from previous estimates of the numbers of stapedius motoneurons and muscle fibers in the cat. One well-labeled stapedius motor axon innervated only a single muscle fiber. In contrast, two labeled axons had over 10 endings and innervated muscle fibers spread over wide areas in the muscle. Two of the axons branched and coursed through two primary stapedius fascicles, indicating that the muscle zones innervated by different primary fascicles are not functionally segregated. In another series of experiments, retrograde tracers were deposited in individual primary nerve fascicles. In every case, labeled stapedius-motoneuron cell bodies were found in each of the physiologically identified stapedius-motoneuron regions in the brainstem. These observations suggest there is little, if any, functional spatial segregation based on separate muscle compartments in the stapedius muscle, despite there being functional spatial segregation in the stapedius-motoneuron pool centrally.

Assessment of vocal function using simultaneous aerodynamic and calibrated videostroboscopic measures.

Despite many attempts to model how vocal fold movements relate to the aerodynamic forces acting on them during phonation, there have been few simultaneous measurements of glottal area and transglottal air pressures and flows. A novel system is described that combines endoscopic measurement of glottal area with aerodynamic flow and pressure measures made during phonation. Results from bench top model tests and from one human subject are presented. For both tests, an aerodynamic model of airflow through a constriction was used to predict the area of the constriction (glottis), and these predictions were compared with endoscopic measurements. The results showed good correlation between predicted and observed areas; however, for small constrictions (<0.025 cm2), whether artificial or glottal, the errors in estimating areas with either optical or aerodynamic methods increase significantly. These results suggest that this measurement system has the potential to enhance the assessment of vocal function.

Innervation of the larynx, pharynx, and upper esophageal sphincter of the rat.

We identified a 'semicircular' compartment of the rat thyropharyngeus muscle at the pharyngoesophageal junction and used the glycogen depletion method to determine how the fibers of this muscle (as well as all others of the pharynx and larynx) are innervated by different cranial nerve branches. The semicircular compartment appears anatomically homologous to the human cricopharyngeus muscle, an important component of the upper esophageal sphincter. While we found very little overlap in the muscle targets of the pharyngeal, superior laryngeal and recurrent laryngeal nerves within the pharynx and larynx, the semicircular muscle receives a dual, interdigitating innervation from two vagal branches: the pharyngeal nerve and a branch of the superior laryngeal nerve we call the dorsal accessory branch. After applying horseradish peroxidase to either of these two nerves, we compared the distribution and number of cells labeled in the brainstem. The dorsal accessory branch conveys a more heterogeneous set of efferent fibers than does the pharyngeal nerve, including the axons of pharyngeal and esophageal motor neurons and parasympathetic preganglionic neurons. The observed distribution of labeled motor neurons in nucleus ambiguus also leads us to suggest that the semicircular compartment is innervated by two subsets of motor neurons, one of which is displaced ventrolateral to the main pharyngeal motor column. This arrangement raises the possibility of functional differences among semicircular compartment motor neurons correlated with the observed differences in brainstem location of cell bodies.

Laser Doppler measurements of intratemporal facial nerve blood flow.

Whereas the anatomy of the vasculature supplying the intratemporal facial nerve is well known, little is known of the dynamics of blood flow within the nerve. The present study was performed to ascertain whether laser Doppler flowmetry (LDF) could detect changes in blood flow within the tympanic segment of the rabbit facial nerve. Compression of the facial nerve immediately distal to the geniculate ganglion resulted in an 80-95 percent reduction in blood flow in the tympanic segment of the nerve, whereas distal neurovascular compression had no effect. Blood flow in the tympanic segment of the nerve fell 40-60 percent during ipsilateral common carotid artery occlusion, but no change occurred with contralateral carotid occlusion. Signal-averaging techniques detected a sinusoidal amplitude modulation of the LDF flow signal that was synchronous with the cardiac cycle. The peak-to-peak amplitude of this modulation was reduced by proximal nerve compression, and the reduction in amplitude was in proportion to the overall reduction in the LDF flow signal. The authors conclude that the direction of blood flow in the tympanic segment of the rabbit facial nerve is primarily proximal to distal. Acute changes in blood flow within the tympanic segment of the nerve could readily be detected using LDF. This technique offers the possibility of monitoring human facial nerve blood flow, and may help elucidate the pathophysiology of various facial neuropathies.

Nonenzymatic Glycosylation of Lepidopteran-Active Bacillus thuringiensis Protein Crystals.

We used high-pH anion-exchange chromatography with pulsed amperometric detection to quantify the monosaccharides covalently attached to Bacillus thuringiensis HD-1 (Dipel) crystals. The crystals contained 0.54% sugars, including, in decreasing order of prevalence, glucose, fucose, arabinose/rhamnose, galactose, galactosamine, glucosamine, xylose, and mannose. Three lines of evidence indicated that these sugars arose from nonenzymatic glycosylation: (i) the sugars could not be removed by N- or O-glycanases; (ii) the sugars attached were influenced both by the medium in which the bacteria had been grown and by the time at which the crystals were harvested; and (iii) the chemical identity and stoichiometry of the sugars detected did not fit any known glycoprotein models. Thus, the sugars detected were the product of fermentation conditions rather than bacterial genetics. The implications of these findings are discussed in terms of crystal chemistry, fermentation technology, and the efficacy of B. thuringiensis as a microbial insecticide.

Acoustic reflex frequency selectivity in single stapedius motoneurons of the cat.

1. The sound frequency selectivities of single stapedius motoneurons were investigated in ketamine anesthetized and in decerebrate cats by recording from axons in the small nerve fascicles entering the stapedius muscle. 2. Stapedius motoneuron tuning curves (TCs) were very broad, similar to the tuning of the overall acoustic reflexes as determined by electromyographic recordings. The lowest thresholds were usually for sound frequencies between 1 and 2 kHz, although many TCs also had a second sensitive region in the 6- to 12-kHz range. The broad tuning of stapedius motoneurons implies that inputs derived from different cochlear frequency regions (which are narrowly tuned) must converge at a point central to the stapedius motoneuron outputs, possibly at the motoneuron somata. 3. There were only small differences in tuning among the four previously described groups of stapedius motoneurons categorized by sensitivity to ipsilateral and contralateral sound. The gradation in high-frequency versus low-frequency sensitivity across motoneurons suggests there are not distinct subgroups of stapedius motoneurons, based on their TCs. 4. The thresholds and shapes of stapedius motoneuron TCs support the hypothesis that the stapedius acoustic reflex is triggered by summed activity of low-spontaneous-rate auditory nerve fibers with both low and high characteristic frequencies (CFs). Excitation of high-CF auditory nerve fibers by sound in their TC "tails" is probably an important factor in eliciting the reflex. 5. In general, the most sensitive frequency for stapedius motoneurons is higher than the frequency at which stapedius contractions produce the greatest attenuation of middle ear transmission. We argue that this is true because the main function of the stapedius acoustic reflex is to reduce the masking of responses to high-frequency sounds produced by low-frequency sounds.

Intracellularly labeled stapedius-motoneuron cell bodies in the cat are spatially organized according to their physiologic responses.

This study examines whether the locations of stapedius-motoneuron cell bodies are correlated with their responses to sound. Single-unit recordings and injections of horseradish peroxidase were made in axons of stapedius motoneurons in the fascicles which run from the facial nerve to the stapedius muscle in the cat. Single units were characterized physiologically by their responses to ipsilateral, contralateral, and binaural sounds. Labeled cell bodies (N = 28) were found in all of the brainstem regions previously identified as containing stapedius motoneurons. Motoneurons characterized as having similar response properties had cell bodies in relatively circumscribed locations. Most (eight of 12) motoneurons excited by sound in either ear had cell bodies in a narrow band around the facial nucleus. Most (seven of eight) motoneurons excited by ipsilateral but not contralateral sound had cell bodies in the cleft between the superior olivary complex and the facial nucleus. All four motoneurons excited by contralateral but not ipsilateral sound had cell bodies located ventromedial to the facial nucleus. The three motoneurons excited only by binaural sound had cell bodies located dorsal to the superior olivary complex. (Two of these were also in the cleft between the superior olivary complex and the facial nucleus.) The cell body of the one motoneuron showing activity in the absence of sound stimulation was located dorsolateral to the facial nucleus. These results show that the cell bodies of stapedius motoneurons with similar electrophysiologic properties tend to have similar locations in the brainstem. The results are consistent with the idea that the stapedius-motoneuron pool is divided into subgroups that are spatially segregated in terms of their patterns of input from the two ears.

Central acoustic tract in an echolocating bat: an extralemniscal auditory pathway to the thalamus.

To determine the sources and targets of auditory pathways that bypass the inferior colliculus in the mustache bat, we injected WGA-HRP in the medial geniculate body and related auditory nuclei of the thalamus as well as in the lower brainstem. We used electrophysiological methods to verify that the injection electrode was in an area responsive to sound. The only thalamic injections that produced retrograde transport to cells in auditory nuclei caudal to the inferior colliculus were those that included the suprageniculate nucleus. These injections labeled a group of large multipolar cells lying between the ventral nucleus of the lateral lemniscus and the superior olivary complex. Neurons in this cell group have also been shown to project to the deep layers of the superior colliculus in the mustache bat. The pathway revealed by these studies is almost identical to the "central acoustic tract" in which fibers course medial to the lateral lemniscus and bypass the inferior colliculus to reach the deep superior colliculus and the suprageniculate nucleus.

The recruitment order of stapedius motoneurons in the acoustic reflex varies with sound laterality.

In many muscles, motor units are recruited in a fixed order with increasing strength of muscular contraction. We show that for the stapedius muscle of the cat, vastly different recruitment orders can be obtained, depending on which ear is acoustically stimulated. The data support the idea that the distribution of inputs to a motoneuron pool can be inhomogeneous and a significant factor in determining recruitment order.

Subcortical connections of the superior colliculus in the mustache bat, Pteronotus parnellii.

The mustache bat, Pteronotus parnellii, depends on echolocation to navigate and capture prey. This adaptation is reflected in the large size and elaboration of brainstem auditory structures and in the minimal development of visual structures. The superior colliculus, usually associated with orienting the eyes, is nevertheless large and well developed in Pteronotus. This observation raises the question of whether the superior colliculus in the echolocating bat has evolved to play a major role in auditory rather than visual orientation. The connections of the superior colliculus in Pteronotus were studied with the aid of anterograde and retrograde transport of wheat germ agglutinin conjugated to HRP. These results indicate that the superior colliculus of Pteronotus is composed almost entirely of the layers beneath stratum opticum. The retinal projection is restricted to a very thin zone just beneath the pial surface. Prominent afferent pathways originate in motor structures, particularly the substantia nigra and the deep nuclei of the cerebellum. Sensory input from the auditory system originates in three brainstem nuclei: the inferior colliculus, the anterolateral periolivary nucleus, and the dorsal nuclei of the lateral lemniscus. The projections from these auditory structures terminate mainly in the central tier of the deep layer. The most prominent efferent pathways are those to medial motor structures of the contralateral brainstem via the predorsal bundle and to the ipsilateral midbrain and pontine tegmentum via the lateral efferent bundle. Ascending projections to the diencephalon are mainly to the medial dorsal nucleus and zona incerta. Thus, the superior colliculus in Pteronotus possesses well-developed anatomical connections that could mediate reflexes for orienting its ears, head, or body toward objects detected by echolocation.

Auditory pathways to the frontal cortex of the mustache bat, Pteronotus parnellii.

In primates, certain areas of the frontal cortex play a role in guiding movements toward visual or auditory objects in space. The projections from auditory centers to the frontal cortex of the bat Pteronotus parnellii were examined because echolocating bats utilize auditory cues to guide their movements in space. An area in the frontal cortex receives a direct projection from a division of the auditory thalamus, the suprageniculate nucleus, which in turn receives input from the anterolateral peri-olivary nucleus, an auditory center in the medulla. This pathway to the frontal cortex bypasses the main auditory centers in the midbrain and cortex and could involve as few as four neurons between the cochlea and the frontal cortex. The auditory cortex is also a major source of input to the frontal cortex. This area of the frontal cortex may link the auditory and motor systems by its projections to the superior colliculus.

Echo intensity compensation by echolocating bats.

When mounted on a swinging pendulum, mustache bats, Pteronotus p. parnellii, emit ultrasonic pulses as they move toward and away from fixed targets. During forward swings they systematically decrease the intensity of their emitted pulses and during backward swings they increase the intensity. In this way, echo strength is continuously adjusted and apparently optimized for signal analysis. We have called this behavior echo intensity compensation. Pteronotus simultaneously Doppler and echo intensity compensate during forward swings of the pendulum but during backward swings they only echo intensity compensate. Pteronotus can regulate the intensity of both the constant frequency and frequency modulated components of their pulses; this regulation is independent of vestibular cues, pulse repetition rates, pulse durations and pulse-echo intervals.

Cochlear microphonic potentials elicited by biosonar signals in flying bats, Pteronotus p. parnellii.

Cochlear microphonic (CM) potentials were recorded from the bat, Pteronotus p. parnellii during tethered flight and during simulated flight on a pendulum. For each emitted signal the frequency of the ca. 61 kHz constant frequency (CF) component was compared with the frequency response characteristics of the animals's ear. The majority of "resting pulses' had CF components with the maximum frequency approximately 200 Hz below the best frequency (BF) of the CM audiogram. Doppler shift compensation occurred only during forward swings of the pendulum and in such a way that the echo CF components were always maintained near the BF, but on the low frequency slope of the CM audiogram. CM responses to emitted pulses were usually small in amplitude and in some animals no responses were seen. Echoes Doppler shifted upward, however, evoked high amplitude potentials. Echo CF components estimated to be at least 43 dB fainter than the emitted pulses evoked higher amplitude CM potentials than the loud emitted pulses. Echoes from large surfaces up to 4.5-5.0 meters away evoked CM potentials as high in amplitude as those elicited by emitted pulses, even when there was no Doppler shift. Beats in the CM were observed on many occasions and occurred as a result of pulse-echo and echo-echo interactions.