Effects of foam rolling on vastus intermedius and lateralis microvascular blood flow.

INTRODUCTION: Foam Rolling (FR) as a technique of self-massage has become a widely used intervention in clinical and sports practice. It is assumed that FR leads to an increased intramuscular microvascular blood flow (MBF), and therefore is commonly recommended as a warm-up or regeneration method. However, no data validate the effects of FR on MBF. This study aimed to assess whether FR increases intramuscular MBF using contrast-enhanced ultrasound (CEUS). METHODS: Ten healthy athletes performed a standardized FR intervention applied to the lateral thigh (3 sets: 45 s FR, 20 s rest). Intramuscular perfusion was determined by CEUS under resting conditions (t0), immediately (t1), and 30 min (t2) after the intervention. Peak enhancement (PE), wash-in rate (WiR), and wash-in perfusion index (WiPI) were evaluated as quantitative perfusion parameters in vastus lateralis (VL) and intermedius (VI) muscle separately via regions of interest mapping. RESULTS: Immediately after the intervention (t1), perfusion parameters showed a non-significant decrease in VL (p = 0.3; PE: -32.1%, WiPI: -29.6%, WiR: -50.4%) and VI (p = 0.4; PE: -10.3%, WiPI: -6.4%, WiR: -35.6%). A non-significant decrease was found at t2 in VL (p = 0.2; PE: -34%, WiPI -33.9%, WiR -61.2%) and VI (p = 0.2; PE -17.6%, WiPI -13.8%, WiR -43.2%). CONCLUSIONS: The common assumption of intramuscular MBF improvement due to FR could not be confirmed for up to 30 min after the intervention. If an increase in intramuscular metabolism or MBF is intended, we recommend that alternative methods (i.e., traditional warm-up) should be preferred.

Density-mediated indirect effects from active predators and narrow habitat domain prey.

The hunting-mode-habitat-domain-range framework suggests that the mechanism driving trophic cascades (i.e., trait-mediated indirect interactions [TMIIs] vs. density-mediated indirect interactions [DMIIs]) should depend upon the functional traits of predators and prey. For example, trophic cascades containing active, broad habitat domain range (BHDR) predators interacting with narrow habitat domain range (NHDR) prey are predicted to arise primarily via TMIIs, because these prey should reduce their conspicuous activity in the presence of these predators. Unfortunately, this hypothesis is difficult to test given the strong bias against studies assessing trophic cascades containing NHDR prey. Furthermore, this hypothesis ignores evidence that (1) active predators can have high consumption rates on prey, (2) continuously responding to active predators foraging across broad areas is energetically costly for prey, and (3) cues from active, BHDR predators may not influence prey density. We examined the TMIIs and total indirect interaction (TII) produced during interactions between an active, BHDR ladybeetle predator (Naemia seriata) and its NHDR prey (scale insects). We exposed scale insects to nonlethal and lethal ladybeetle predators in laboratory mesocosms for 15 weeks. We measured the growth of the scale insect's host plant (cordgrass) and the population density of scale insects. Contrary to theory, nonlethal ladybeetles did not induce TMIIs. However, lethal ladybeetles increased cordgrass total and root dry biomass by 36% and 44%, respectively, suggesting the presence of strong DMIIs. Additionally, both lethal and nonlethal ladybeetles reduced scale insect population density. Our findings suggest that DMIIs, rather than TMIIs, can result from interactions between active BHDR predators and NHDR prey.

[Laryngeal and larynx-associated reflexes]. / Larynxreflexe und larynxassoziierte Reflexe.

The laryngeal adductor reflex and the pharyngoglottal closure reflex protect the trachea and lower respiratory tract against the entrance of foreign material. The laryngeal expiration reflex and the cough reflex serve to propel foreign material, which has penetrated in the cranial direction. The inspiration reflex, the sniff reflex, and the swallowing reflex are further larynx-associated reflexes. In patients with dysphagia the laryngeal adductor reflex can be clinically tested with air pulses. The water swallow test serves to show the integrity of the cough reflex. The sniff reflex is useful to test the abduction function of the vocal folds. Future studies should address laryngeal reflexes more specifically, both for a better understanding of these life-supporting mechanisms and to improve diagnostic procedures in patients with impaired laryngeal function.

[Deliberate release of the laryngeal adductor reflex via microdroplet impulses: Development of a device]. / Gezielte Auslösung des laryngealen Adduktionsreflexes per Mikrotröpfchenabgabe: Entwicklung einer Vorrichtung.

BACKGROUND: The laryngeal adductor reflex (LAR), a reflexive vocal fold closing mechanism, includes an early, probably di- or oligosynaptic ipsilateral LAR1- and a late ipsilateral and contralateral LAR2 polysynaptic component. In a clinical evaluation of dysphagia the LAR can be triggered by air pulses or tactile stimuli and typically assessed only qualitatively. METHODOLOGY: The development and construction of a device that can selectively shoot very small water droplets (microdroplet impulse testing MIT). RESULTS: The MIT device has a water reservoir with an infinitely adjustable pressure. The opening period of the piezo-electrically operated valve determines the droplet size. With a high-speed camera system, the change in the airspeed of the drop can be determined, depending on the set water reservoir pressure. With the knowledge of the droplet size, the shooting speed and the estimation of the distance between the valve and laryngeal mucosa or airspeed can be determined the muzzle energy. By mounting the MIT device to a high speed glottography system, the time between the impact of the droplet on the laryngeal mucosa and the start of the laryngeal adduction, the LAR latency can be determined using an image by image evaluation. DISCUSSION: In dysphagia with penetration or aspiration it is presumed that the protective function of the larynx is no longer adequately ensured. The MIT-LAR device provides a valid and reliable method to assess LAR quantitatively. Furthermore, it holds the promise of being a simple to handle method that can be used clinically for routine diagnostics.

[Characterization of the Laryngeal Adductor Reflex by Stimulation with Microdroplets Impulses (Microdroplet Impulse Testing)]. / Charakterisierung des laryngealen Adduktionsreflexes durch Stimulation mit Mikrotröpfchen-Impulsen (microdroplet impulse testing).

BACKGROUND: The larynx is considered a crossing point between breathing and swallowing pathways. During swallowing, the airway below the glottis must be protected against food components by an appropriate laryngeal closure mechanism. The laryngeal adductor reflex (LAR) with an early, probably di- or oligosynaptic interconnected ipsilateral LAR1- and a late ipsilateral and contralateral LAR2 polysynaptic component is believed to serve as such a mechanism. Here we aimed to measure and characterize the LAR in healthy volunteers and to compare the data obtained with previously published data. METHODS: We designed a prospective pilot study. 10 healthy volunteers (22-57 years) participated. To elicit the LAR we used a newly designed microdroplet impulse testing (MIT) device: very small waterdroplets were shot onto the endolaryngeal mucosa. By simultaneously observing the anatomical structures with a high speed glottography system, the time between impact of the microdroplet on the mucosa and the beginning of the adduction movement and thus an approximate value for the reflex latency could be determined. RESULTS: An early adduction movement corresponding to LAR1 could not be detected. The measured LAR2 latency time was higher than the EMG LAR2 data. No significant latency difference between right and left stimulation was found. DISCUSSION: Since we were unable to demonstrate any LAR1 component it may be that muscle activity observable by EMG may not be sufficient to lead to a visible medial vocal cord movement. The longer LAR2 latency compared to EMG data may be explained by the fact that the visually vocal cord movement occurs after a delay although muscle activity already started as evidenced by EMG.Further studies on LAR are warranted, especially since our results also raise questions about the clinical significance of the LAR.

Intramedullary nailing of trochanteric fractures: central or caudal positioning of the load carrier? A biomechanical comparative study on cadaver bones.

BACKGROUND: Current recommendations with regard to central or caudal positioning of the femur head carrier in the management of trochanteric fractures are contradictory. METHODS: A standardised pertrochanteric osteotomy was stabilised in 15 pairs of cadaver femurs by means of intramedullary osteosynthesis (5xPFN-A-Synthes, 5xIntertan-Smith&Nephew, 5xTargon-PF-Aesculap). For each pair randomised central (group A) or caudal (group B) implantation of the femoral neck component was performed. Subsequently, the constructs were axially loaded to 2100N. In the absence of cut out after 20,000 cycles, load was increased to a maximum force of 3100N. Angular displacement was recorded based on ultrasound. Migration of the load carrier in the femoral head was monitored radiologically. FINDINGS DISPLACEMENT: No significant difference between groups (p>0.15) was found for the first 50 load cycles. A significantly greater degree of varus deformity was observed in group A (p=0.049) after 2000 load cycles and became more apparent as the number of load cycles increased (after 6000 cycles p=0.039, after 20,000 cycles p=0.034, after 22,000 cycles p=0.016). Angular displacement in the other two planes did not differ significantly across groups. CUT OUT: Migration of the load carrier in the femoral head was not significantly different for the two groups. Overall cut out occurred in 9 constructs, 3 in group A and 6 in group B. The difference in cut-out rate was not significant (p=0.213, chi-squared test). CONCLUSION: Biomechanical superiority can be shown for caudal positioning of the femoral neck load carrier in terms of reduced varus deformity. The incidence of cut out is however unaffected by the position of the load carrier.

[Comparison of Rebound tonometry versus Perkins tonometry in the supine glaucoma patient]. / Vergleich zwischen Rebound-Tonometrie und Perkins-Tonometrie am liegenden Glaukompatienten.

BACKGROUND: The aim of this study was to evaluate the concordance of intraocular pressure (IOP) measurements in the supine glaucoma patient using a Perkins applanation tonometer (PAT) compared to the iCare® rebound tonometer (RT), a hand-held device requiring no local anaesthesia. METHODS: 73 left eyes of 73 glaucoma patients were included in this consecutive case study and measured both by supine Perkins applanation tonometry and by right lateral posture rebound tonometry in the supine position (RLP). The patients were divided into three subgroups dependent on IOP (SG-1: 0 - 15 mmHg, SG-2: 16 - 22 mmHg, SG-3: more than 23 mmHg). RESULTS: The mean deviation between RT and PAT was 2.6 ± 4.0 mmHg, the 95 % confidence interval was -5.3 to 10.4 mmHg. 69 % of the measurements showed deviations within 3 mmHg between the two devices. Deviation was smallest in SG-2, and largest in SG-3. CONCLUSIONS: Rebound tonometry is comfortable to use even in supine patients. RT measurement agreed overall significantly with those of Perkins applanation tonometry, generally overestimating PAT measurement. In high IOP values, RT did not correlate as well with PAT as in moderate IOP levels.

[Traumatic lumbosacral dislocation - an underrated injury]. / Traumatische lumbosakrale Spondylolisthesis - eine unterschätzte Verletzung.

Traumatic lumbosacral dislocations are rare. We report two cases with initially missed posttraumatic lumbosacral dislocations. The reported cases and the review of the literature show that, especially, accident victims with multiple fractures of the lumbar transverses processes may require a CT scan to confirm fractures or dislocations of L5/S1. Follow-up examinations due to persisting pain after physiotherapy should include lateral X-rays of the lumbar spine of the patient standing. According to the literature and our experience, the treatment of traumatic lumbosacral dislocation usually consists of open reduction and postero-lateral or dorso-ventral fusion of the unstable segments.

Ultrastructural localization of the serotonin transporter in superficial and deep layers of the rat prelimbic prefrontal cortex and its spatial relationship to dopamine terminals.

Dopamine levels within the prefrontal cortex (PFC) can be manipulated by selective inhibitors of the serotonin transporter (SERT). However, the cellular mechanisms underlying these effects are not clear. The present study sought to examine the distribution of immunogold-silver labeling for SERT (SERT-ir) in the rat prelimbic PFC and to describe its ultrastructural spatial relationship to dopamine axons labeled by immunoperoxidase staining for tyrosine hydroxylase (TH-ir). SERT was localized to axonal profiles that ranged in size from fine caliber fibers containing dense SERT-ir, primarily along the membrane, and rarely forming synapses to large, spherical varicosities exhibiting less dense staining, mainly within the cytoplasm, and more commonly forming synapses. Synaptic contacts of SERT profiles were typically asymmetric, axospinous, and more frequent in superficial (38%) than deep (19%) layers. For TH-ir profiles, only 24% were within the same 13.8 microm(2) microenvironment as SERT-ir profiles. Furthermore, TH-ir and SERT-ir profiles were rarely directly apposed to each other or convergent onto common dendritic structures. Instead, these two profiles were typically separated by an average distance of 1.30 microm in the coronal plane, a value that did not vary with the size of SERT-ir axons, the amount of SERT labeling, or the cortical layer examined. These results are consistent with two populations of SERT profiles within the rat prelimbic PFC that may arise from different raphe nuclei or that represent varicose and intervaricose portions of the same axons. Moreover, the functional interactions between cortical serotonin and dopamine systems that may contribute to the therapeutic efficacy of antidepressant drugs are likely to occur over distances greater than 1 microm.

Immunolocalization of the cocaine- and antidepressant-sensitive l-norepinephrine transporter.

Norepinephrine (NE) transporters (NETs) constitute the primary mechanism for inactivation of synaptically released NE, are targets for multiple antidepressants and psychostimulants, and have been reported to be deficient in affective and autonomic disorders. Although the regional distribution of NETs has been defined through synaptosomal transport and autoradiographic approaches, NET protein expression has yet to be characterized fully in the central nervous system (CNS). We identified a cytoplasmic NET epitope (amino acids 585-602) and corresponding antibody (43411) that permits cellular localization of endogenous NET expression in the CNS and periphery. In the adult rat brain, NET labeling was confined to noradrenergic neuronal somata, axons, and dendrites, including extensive arborizations within the hippocampus and cortex, but was absent from epinephrine- and dopamine-containing neurons. Intracerebroventricular anti-dopamine beta-hydroxylase/saporin, a treatment that destroys a majority of noradrenergic neurons and their projections, validated the specificity of the 43411 antibody. At the level of light microscopy, 43411 labeling colocalized with the axonal markers syntaxin, synaptophysin, and SNAP-25. Indirect immunofluorescence revealed a nonuniform pattern of NET expression along axons, particularly evident within sympathetic fibers of the vas deferens, reflecting a high degree of spatial organization of NE clearance. NET labeling in somata was intracellular and absent from plasma membranes. Among nonneuronal cells, glial cells lacked NET immunoreactivity, whereas CNS ependymal cells were an unexpected site of labeling. NET immunoreactivity was also evident in a subset of adrenal chromaffin cells where labeling appeared to be predominantly associated with intracellular vesicles. Initial ultrastructural evaluation via preembedding immunogold techniques also revealed substantial cytoplasmic NET immunoreactivity in axon terminals within the prelimbic prefrontal cortex, consistent with postulates of regulated trafficking controlling neurotransmitter clearance. NET visualization should be of significant benefit in evaluating neuronal injury resulting from chronic drug exposure and in disease states.

Acute regulation of norepinephrine transport: II. PKC-modulated surface expression of human norepinephrine transporter proteins.

Norepinephrine (NE) transporters (NETs) found in the neuronal plasma membrane mediate the removal of NE from the extracellular space, limiting the activation of adrenoceptors at noradrenergic synapses. Our previous studies with the noradrenergic neuroblastoma SK-N-SH have revealed a muscarinic receptor-triggered regulation of NET surface density and transport capacity, mediated in part by protein kinase C activation. Low abundance of NET proteins in this native cell model, however, preclude direct confirmation of altered trafficking of NET proteins. In our study, we monitored the activity and surface distribution of human NET proteins in transient and stably-transfected cell lines after application of kinase activators and inhibitors. Using hNET stably transfected HEK-293 and LLC-PK1 cells, as well as transiently transfected COS-7 cells, we demonstrate that PKC-activating phorbol esters, beta-PMA or beta-PDBu selectively diminish l-NE transport capacity (Vmax) with little change in NE Km. Effects of phorbol esters are rapid, stereospecific and blocked by protein kinase C inhibitors, staurosporine and bisindolylmaleimide I. As in SK-N-SH cells, beta-PMA induces a reduction in intact cell [3H]nisoxetine binding sites with no change in nisoxetine Kd or total membrane NET density. Cell-surface biotinylation and confocal immunofluorescence techniques confirm that protein kinase C-dependent reductions in NE transport capacity and whole-cell antagonist binding density are accompanied by reductions in cell-surface human NET protein expression. Together these findings argue for kinase-modulated protein trafficking as a potential route for acute regulation of antidepressant-sensitive NE clearance.

Regulated phosphorylation and trafficking of antidepressant-sensitive serotonin transporter proteins.

Presynaptic serotonin (5-hydroxytryptamine, 5-HT) transporters (SERTs) mediate antidepressant-sensitive clearance of 5-HT following release. Although we have been aware for decades that SERT-mediated 5-HT clearance can be modulated by exogenous agents including serotonin-selective reuptake inhibitors, amphetamines, and cocaine, we have had little reason to speculate that SERT activity was actively controlled through endogenous pathways. Recent studies indicate that SERTs are likely to be trafficked to specific plasma membrane subdomains to achieve localized clearance of 5-HT, and that the number of SERTs resident in the plasma membrane is controlled through kinase- and phosphatase-linked pathways. In particular, roles for protein kinase C and phosphatase 2A become apparent through studies with enzyme activators and inhibitors in SERT-transfected cells, where SERT proteins are rapidly phosphorylated in parallel with transporter redistribution and loss of functional uptake capacity. Based on our findings, and the studies of others in native tissues and transfected cells, we propose a model whereby SERTs are organized in a macromolecular complex in the plasma membrane that may serve to locate reuptake activity near release sites. Although many elements of this model remain hypothetical, our findings suggest a much more dynamic picture of transporter-mediated 5-HT reuptake than typically described and suggest opportunities both for the development of new SERT regulatory agents and for the identification of regulatory pathways that may be compromised in mental illness.

Polarized expression of the antidepressant-sensitive serotonin transporter in epinephrine-synthesizing chromaffin cells of the rat adrenal gland.

Antidepressant-sensitive serotonin (5-hydroxytryptamine, 5HT) transporters (SERTs) clear the amine from extracellular spaces in the CNS and periphery as a mechanism for transmitter inactivation and recycling. Although it is known that SERTs are preferentially expressed on basolateral domains in transfected epithelial cells, details of the transporter's membrane localization in vivo are lacking. 5HT and 5HT receptors have been identified in the rodent adrenal gland. Using SERT antagonist autoradiography, we establish the presence of antidepressant-sensitive transport sites in the rat adrenal medulla. Immunofluorescence experiments using antibodies specific for the SERT COOH and NH2 termini, for 5HT, or for catecholamine biosynthetic enzymes suggest that SERT mediates intra-cellular 5HT accumulation by epinephrine-secreting chromaffin cells. Using confocal microscopy, we establish that SERT expression is nonuniformly distributed along the plasma membrane of chromaffin cells. Notably, SERT immunoreactivity is largely absent from plasma membranes bordering smooth muscle that surrounds vascular sinusoids. Rather, SERT is highly expressed in membranes adjoining other chromaffin cells, consistent with a role for 5HT and SERT in autocrine or paracrine control of chromaffin cell physiology. SNAP-25, a t-SNARE protein implicated in neurotransmitter release, was found to colocalize with SERT. In contrast, Na,K ATPase and NCAM are uniformly distributed along the entire perimeter of chromaffin cell membranes. These findings underscore a role for 5HT and SERT in adrenal physiology, reveal unrecognized polarity of chromaffin cell plasma membranes, and warrant a consideration of common targeting mechanisms localizing amine transporters near release sites.

Cloning and expression of the mouse serotonin transporter.

A mouse brain cDNA encoding the high-affinity serotonin transporter (SERT) has been identified and characterized. The mouse transporter sequence (mSERT) encodes a protein of 630 amino acids which contains twelve potential transmembrane domains (TMDs), N-linked glycosylation and kinase-mediated phosphorylation sites, and high levels of homology with rat and human SERTs. Heterologous expression of mSERT in COS-I cells resulted in a [3H]serotonin transport activity characterized by kinetic saturability (Km = 403 +/- 42 nM. Vmax = 1.02 +/- 0.10 pmol/mg/min), Na1 and Cl- dependences (5HT:Na+:Cl- coupling ratio of 1:1:1), and sensitivity to known inhibitors of serotonin transport (including antidepressant and psychostimulant agents). Northern analysis using mSERT cDNA as probe revealed a single 3.4 kb mRNA species expressed in mouse lung, midbrain and brainstem regions, and absent from heart and liver. In situ hybridization studies further established the specific localization of mSERT gene expression to the raphe nuclei of the mouse midbrain. The identified mSERT cDNA sequence provides a new tool for the evaluation of serotonin transport pharmacology in heterologous expression systems and provides an opportunity for the evaluation of mSERT gene expression in a well-characterized model of mammalian development.

The myelopoietic effects of a Serratia marcescens-derived biologic response modifier in a mouse model of thermal injury.

The proliferative defects observed in phagocytic stem cells after major thermal injuries may be caused by an inadequate production of colony-stimulating factors (CSFs), a family of hemopoietic cytokines necessary for the production and function of granulocytes and monocytes. In this study a biologic response modifier (S-BRM) consisting of sized vesicles derived from the cell membrane and ribosomes of Serratia marcescens was investigated in a mouse model of thermal injury to determine its ability to augment postburn myelopoiesis. Treatment of burned mice with S-BRM was well tolerated and was associated with statistically significant increases in absolute numbers of circulating granulocytes and monocytes compared with burned mice receiving saline solution. In addition, the size of the splenic myeloid stem cell compartment, as measured by granulocyte-macrophage stem cell colony formation in soft agar, was markedly expanded. Finally, plasma levels of CSF were increased significantly in burned mice receiving S-BRM but were not elevated in burned littermates treated with saline solution. These data suggest that production of CSF is suboptimal after thermal injury and S-BRM is capable of up-regulating postburn myelopoiesis by causing the release of CSF into the systemic circulation.

Spatial and temporal patterns of muscle cleavage in the chick thigh and their value as criteria for homology.

Regions of lower cell density, called cleavage zones, emerge within the dorsal and ventral muscle masses in the vertebrate limb to separate distinct muscles. In the chick thigh, the stereotyped patterns of separation have been broadly outlined, but differences in interpretation exist because no criteria for separation have been defined, and the tissues of the limb are indistinct early in development. We have examined the cleavage process using modern applications of light microscopy and immunocytochemistry to completely detail the spatial and temporal progression of cleavage in stage 27-32 embryos. We find that each muscle has a complex but characteristic pattern of separation along the proximodistal axis. The complex pattern of separation is not related to the positions of muscles within the thigh, locations of blood vessels, activity patterns of muscles, or innervation patterns. The initial separation patterns are more straightforward than later separations and may be of value in determining the phylogenetic history of limb muscles since the same patterns are common to many tetrapods. Our detailed documentation clarifies the ontogeny of the thigh musculature and reveals more complex separation patterns between muscles than previously described.

Ultrastructural and morphometric analysis of the separation of two thigh muscles in the chick.

Limb muscles separate from one another in a complex but highly stereotyped sequence and spatial pattern. The process of separation is characterized by the progression of a region of increased extracellular space, the cleavage zone, along the proximodistal axis between the individual muscle anlagen. We analyzed ultrastructurally the muscles and cleavage zone during the separation of two representative muscles, the developing sartorius and iliotibialis in the chick thigh, to establish an accurate baseline for an analysis of the mechanisms of separation. Comparisons of the morphology and distribution of cells before and after separation show no evidence that muscles became separated by the massive influx of an exterior cell population; if populations invade the cleavage zone, they are small. We do find characteristic transitions within the cell population of the cleavage zone in situ that could accomplish cleavage without invoking massive cell movements. These progressive transitions within the cleavage zone include a loss of close cell-cell interactions, an increase in extracellular space, the assumption of a more stellate morphology by mesenchyme cells, and a gradual alteration in the composition of the extracellular matrix from one typical of early muscle to one typical of loose connective tissue. Myotubes do differentiate between the incipient muscles, ruling out the possibility that the location where muscles will separate is defined by sites where myotubes fail to differentiate. Instead, the myotubes in the cleavage zone gradually diminish in number and appear to be specifically recognized and removed from the cleavage zone by phagocytes. We suggest that the transitions within the cleavage zone, including the loss of muscle cells, are a result of the progressive differentiation of loose connective tissue. If so, then the spatial pattern and process of cleavage is a consequence of spatially programmed cell differentiation.