Sputum proteomic signature of gastro-oesophageal reflux in patients with severe asthma.

Gastro-oesophageal reflux disease (GORD) has long been associated with poor asthma control without an established cause-effect relationship. 610 asthmatics (421 severe/88 mild-moderate) and 101 healthy controls were assessed clinically and a subset of 154 severe asthmatics underwent proteomic analysis of induced sputum using untargeted mass spectrometry, LC-IMS-MSE. Univariate and multiple logistic regression analyses (MLR) were conducted to identify proteins associated with GORD in this cohort. When compared to mild/moderate asthmatics and healthy individuals, respectively, GORD was three- and ten-fold more prevalent in severe asthmatics and was associated with increased asthma symptoms and oral corticosteroid use, poorer quality of life, depression/anxiety, obesity and symptoms of sino-nasal disease. Comparison of sputum proteomes in severe asthmatics with and without active GORD showed five differentially abundant proteins with described roles in anti-microbial defences, systemic inflammation and epithelial integrity. Three of these were associated with active GORD by multiple linear regression analysis: Ig lambda variable 1-47 (p = 0·017) and plasma protease C1 inhibitor (p = 0·043), both in lower concentrations, and lipocalin-1 (p = 0·034) in higher concentrations in active GORD. This study provides evidence which suggests that reflux can cause subtle perturbation of proteins detectable in the airways lining fluid and that severe asthmatics with GORD may represent a distinct phenotype of asthma.

Temperature-dependent global gene expression in the Antarctic archaeon Methanococcoides burtonii.

Methanococcoides burtonii is a member of the Archaea that was isolated from Ace Lake in Antarctica and is a valuable model for studying cold adaptation. Low temperature transcriptional regulation of global gene expression, and the arrangement of transcriptional units in cold-adapted archaea has not been studied. We developed a microarray for determining which genes are expressed in operons, and which are differentially expressed at low (4°C) or high (23°C) temperature. Approximately 55% of genes were found to be arranged in operons that range in length from 2 to 23 genes, and mRNA abundance tended to increase with operon length. Analysing microarray data previously obtained by others for Halobacterium salinarum revealed a similar correlation between operon length and mRNA abundance, suggesting that operons may play a similar role more broadly in the Archaea. More than 500 genes were differentially expressed at levels up to ≈ 24-fold. A notable feature was the upregulation of genes involved in maintaining RNA in a state suitable for translation in the cold. Comparison between microarray experiments and results previously obtained using proteomics indicates that transcriptional regulation (rather than translation) is primarily responsible for controlling gene expression in M. burtonii. In addition, certain genes (e.g. involved in ribosome structure and methanogenesis) appear to be regulated post-transcriptionally. This is one of few experimental studies describing the genome-wide distribution and regulation of operons in archaea.

Activity patterns of proteasome subunits reflect bortezomib sensitivity of hematologic malignancies and are variable in primary human leukemia cells.

Proteasomal proteolysis relies on the activity of six catalytically active proteasomal subunits (beta1, beta2, beta5, beta1i, beta2i and beta5i). Applying a functional proteomics approach, we used a recently developed activity-based, cell-permeable proteasome-specific probe that for the first time allows differential visualization of individual active proteasomal subunits in intact primary cells. In primary leukemia samples, we observed remarkable variability in the amounts of active beta1/1i-, beta2/2i- and beta5/5i-type of subunits, contrasting with their constant protein expression. Bortezomib inhibited beta5- and beta1-type, but to a lesser extend beta2-type of subunits in live primary cells in vitro and in vivo. When we adapted the bortezomib-sensitive human acute myeloid leukemia cell line HL-60 to bortezomib 40 nM (HL-60a), proteasomal activity profiling revealed an upregulation of active subunits, and residual beta1/beta5-type of activity could be visualized in the presence of bortezomib 20 nM, in contrast to control cells. In a panel of cell lines from hematologic malignancies, the ratio between beta2-type and (beta1 + beta5)-type of active proteasomal polypeptides mirrored different degrees of bortezomib sensitivity. We thus conclude that the proteasomal activity profile varies in primary leukemia cells, and that the pattern of proteasomal subunit activity influences the sensitivity of hematologic malignancies toward bortezomib.

[Long-term follow-up after surgery for thoracic outlet syndrome (TOS)]. / Langzeitresultate nach operativer Therapie bei Thoracic-outlet-Syndrom (TOS).

This is a long-term follow-up analysis of patients who have been operated on for Thoracic Outlet Syndrome (TOS) at our clinic in order to evaluate the quality of therapy and the criteria of indications for surgery. 39 patients with a total of 45 surgical procedures were examined after a median follow-up of 8.8 years. The results in this study are based exclusively on the subjective outcome assessment by the patients themselves. Assessment of the long-term result in the "vascular TOS" group (13 cases = 29 %) was good in ten cases (77 %), fair in two cases (15 %) and poor in one case (8 %). In agreement with the literature, we were able to achieve the best results in this group. In the "true neurological TOS" group (28 cases = 62 %), assessment of the long-term result was good in 19 cases (68 %), fair in six cases (21 %) and poor in three cases (11 %). A clear tendency to a poor prognosis could be seen in women with a combination of cervical rib and fibrous band and a long delay between onset of symptoms and surgery. Assessment of long-term result in the "disputed TOS" group (four cases = 9 %) showed good results in three cases and a fair result in one case. In the absence of objective pathologies, only few and carefully selected patients were operated upon. The presented long-term results confirm the use of individual therapeutic concepts with special consideration of anatomy and clinical picture.

["True neurologic thoracic outlet syndrome" -- anatomical features and electrophysiological long-term follow-up of lateral thenar atrophy]. / "True Neurologic Thoracic Outlet Syndrome" -- Anatomie und elektrophysiologischer Langzeitverlauf bei lateraler Thenaratrophie.

Atrophies of the intrinsic muscles of the hand are considered to be a typical symptom of the "true neurologic" form of thoracic outlet syndrome (TOS). The classical form of this entity was described as early as 1970, consisting of a cervical rib or a prolonged transverse process of C7, complete with a fibrous band to the first thoracic rib, resulting in atrophy of the intrinsic muscles of the hand. All our TOS patients presenting with such atrophy displayed anatomical findings consistent with this definition. Based on this observation, the TOS classification currently in clinical use, which differentiates between "disputed" and "true neurologic" subgroups of the neurologic form, is reviewed. In all cases of "true neurologic TOS" with atrophy of the intrinsic muscles of the hand, the lateral thenar muscles are affected first. We present the electrophysiological long-term results of such thenar atrophies of seven patients with eight operated extremities after brachial plexus decompression. The amplitude of the neurographically measured potential over the opponens pollicis and the abductor pollicis brevis muscle, respectively, was defined as quantitative parameter for muscles atrophy. Neither distinct reinnervation nor progressive denervation was evident in any of the cases after a follow-up period, on average, of more than five years post surgery. These findings are in conflict with clinical observations reporting a major postoperative improvement of the motor deficits.

[Verification of thumb-specific muscles in a triphalangeal thumb with magnetic resonance imaging]. / Nachweis daumenspezifischer Muskulatur beim triphalangealen Daumen mittels MRT.

The presence of three phalanges in the first digit is considered to be a relatively rare congenital hand malformation. Six groups of this deformity can be distinguished: some digits are opposable, others resemble a non-opposable five-fingered hand. In cases of a hypoplastic thenar region with a restrained opposition, a clear verification of thumb-specific musculature has been hardly possible. We report of the possibility of a non-invasive identification of thumb-specific muscles by means of magnetic resonance imaging.

Compression neuropathy of the radial palmar thumb nerve.

Compression neuropathy of a single digital nerve is a rare entity. We report the case of a patient with numbness in the distribution of the radial digital nerve of the thumb caused by the use of a walking stick. The nerve was compressed between the handle of the stick, the loop and the radial sesamoid bone of the first metacarpophalangeal joint. The site of the lesion was confirmed by electrophysiologic examination. Orthodromic recording of the sensory response from the radial palmar digital nerve of the thumb documented a complete absence of nerve action potential whereas the ulnar digital thumb nerve showed a normal response. Sensory function was restored when a padded ski glove was used to protect the area of the metacarpophalangeal joint whilst using the stick.

[Wartenberg's Sign of Ulnar Nerve Lesion. A Contribution to Pathophysiology and to the Differential Diagnosis]. / Das Wartenberg-Zeichen des Nervus ulnaris. Ein Beitrag zur Pathophysiologie und Differenzialdiagnose1.

The abduction stance of the small finger is frequently, but not necessarily due to ulnar nerve paresis. Five cases suffering from bothersome permanent abduction of the small finger and referred under the diagnosis of ulnar nerve paresis are presented. Clinical, electrodiagnostic and imaging evaluation revealed different causes. While partial paresis with the function of the abductor digiti minimi muscle preserved usually results in a disturbing abduction stance, complete paresis of the ulnar nerve causes a less severe abduction posture of the small finger. Operative measures are indicated when the stance of the small finger is disturbing and when sufficient time has passed to make sure that spontaneous recovery cannot be presumed. Clinical, electrodiagnostic and imaging evaluation of three neurogenic cases disclosed a lesion of the ramus profundus distal to the branches innervating hypothenar muscles in one case, ulnar nerve injury with neuromuscular hyperactivity of the abductor digiti minimi muscle following split repair in another case and syringomyelia in the third case. Two patients revealed an abduction posture of the little finger of non-neurogenic origin. One of them showed closed ligament injuries. The other patient revealed necrosis, scarring and contracture of hypothenar muscles and atrophy of the third palmar interosseous muscle following compression in a tight cast.

[Methods, indications and validation of intraoperative nerve conductivity testing]. / Methode, Indikationen und kritische Wertung der intraoperativen Nervenfunktionsdiagnostik*

During nerve surgery, electrodiagnostic methods are applied to assess the availability and viability of nerve fibers and to adjust operative measures accordingly. The validity of this procedure is verified by histology and by the outcome of the operation. This paper explains the techniques of intraoperative nerve action potential (NAP) and somatosensory evoked potential (SEP) recording, how to interpret the electrodiagnostic results, and describes both the special features and the limitations of the methods. We found reliable results of neurography, detecting the presence or absence of spontaneous nerve regeneration across a lesion in continuity months before the reinnervation reached its final target. Based on our results, we suggest that NAP recording of the exposed nerve can widely prevent unnecessary nerve or fascicle resection. Besides this important indication, the nerve function evaluation was successfully used in nerve surgery whenever the quality of the nerve parenchyma was crucial to the operative management. Further indications such as evaluating brachial plexus lesions and the condition of nerve roots, judging the proximal coaptation site in nerve reconstruction, tracing the site of a nerve lesion and identifying the pathophysiology of nerve malfunction are exemplified. Intraoperative nerve conductivity testing should not be considered as a replacement of but rather as a complement to preoperative clinical, electrophysiological and imaging evaluations and a thorough intraoperative morphological examination.

Tattooing agents for nerve marking in experimental surgery.

Experimental nerve surgery involves test procedures, including those for nerve lesions in continuity, that leave no visible traces of impairment after surgery. In such cases, non-resorbable sutures are usually used to mark the lesion sites on the nerves. However, this method has two drawbacks: it is not completely atraumatic, and may be frustrating due to displacement of the suture material. The authors demonstrate the use of carbon tattoo pigment to mark nerve lesions permanently, thus allowing their identification reliably at any later date. Following successful preliminary experiments, the tattooing procedure was used in 12 New Zealand White rabbits that had been operated on for a specific nerve regeneration problem. Altogether, 56 tattoo marks were set. The small pigment spots were well-preserved and clearly visible during a second and third operation 4 and 15 weeks later. Histologic examination identified the carbon granules in the outer epineurium; there were no signs of inflammation. This simple, atraumatic, inert, and permanent method for nerve markings in the experimental animal is recommended.

Cervical outlet syndrome due to an accessory part of the trapezius muscle in the posterior triangle of the neck.

A 48-year-old man presented with pain and sensory impairment radiating from the neck to the thumb and forefinger of the right hand when lifting weights and turning or tilting the head. The symptoms were due to an anomalous accessory part of the trapezius muscle crossing the upper part of the brachial plexus. Excision of the anomalous muscle and release of the clavicular part of the sternocleidomastoid muscle abolished the complaints.

[Effective treatment of a large muscle hernia by local botulinum toxin administration]. / Wirksame Entlastung einer grossen Muskelhernie durch lokale Botulinumtoxin-Anwendung.

A 53-year-old patient developed an impairing muscle hernia when a fascia lata graft was harvested as a substitute for a cruciate ligament of the knee and closure of the defect was not possible. The fascial defect enlarged with time, extending along the whole upper leg. The large muscle protrusion and incarceration in the distal fascial slit was extremely painful during walking and getting up from a chair. Since autologous grafts were disregarded because of the high tissue pressure and alloplastic substitutes seemed problematic, the large hernia was successfully reduced by local muscle denervation with injections of botulinum-A toxin into the protruding vastus lateralis muscle. This procedure achieved relief of pain and enabled the patient to walk without complaints. Side effects were not observed.

[Skin symptoms in neurological diseases]. / Hautsymptome bei neurologischen Erkrankungen.

The considerable variety of diseases involving both the skin and the peripheral or central nervous system is due to the developmental relationship between skin and nervous system. Diseases of the nervous system resulting in skin changes comprise disorders of nails, hair, sweat and sebaceous glands as well as neuropathic ulcers in various manifestations. Neurocutaneous syndromes (phacomatoses) are nevoid systemic diseases comprising neurofibromatoses, tuberose sclerosis, ataxia telangiectasia, nevoid angiomatoses and other neurocutaneous disorders. Polytopic disorders of the skin and nervous system may be due to infectious diseases, diseases of the connective tissue, fat, inflammatory vascular and granulomatous disorders.

[Nerve compression syndromes of the extremities]. / Nervenkompressions-syndrome der Extremitäten.

Peripheral entrapment neuropathies occur in high frequency and present clinically with a wide range of variations. They need to be recognized early enough in order to initiate correct therapy and so to obviate serious nerve lesions and possible neurological sequelae. This paper overviews the essentials of the compression neuropathies as they are encountered in both upper and lower extremities. Pathomechanisms , pathogenesis, evaluation considerations as well as differential diagnosis and basic treatment algorithms are emphasized.

Interactions of Lyn with the antigen receptor during B cell activation.

Signaling through the B cell antigen receptor requires a complex set of interactions involving transmembrane components of the IgM receptor complex and cytosolic protein-tyrosine kinases. We have focused on the nature of these protein-protein interactions, the requirements for their occurrence, as well as the temporal sequence of events during the activation process. We found that cross-linking B cell antigen receptors at 0 degree C resulted in the rapid association of the Src-family protein-tyrosine kinase, Lyn, with the antigen receptor complex as judged by the presence of Lyn in anti-IgM and anti-phosphotyrosine immune complexes and the presence of MB-1 in anti-Lyn immune complexes. Receptor engagement also resulted in the rapid association of Lyn with the phosphotyrosine phosphatase, CD45. This association of Lyn with receptor components was stable in the detergent Brij 96, but was readily disrupted by Nonidet P-40, suggesting the involvement of hydrophobic interactions in stabilizing formation of the Lyn-receptor complex. The protein-tyrosine kinase, Syk, was also found associated with activated receptor complexes. This association of Syk with components of the antigen receptor complex was stable to Nonidet P-40. Antibodies directed against the carboxyl teminus of Syk, but not against the amino-terminal SH2 domain, co-immunoprecipitated MB-1 from activated cells, consistent with the binding of Syk through an SH2 domain-phosphotyrosine interaction.

Inhibition of mast cell Fc epsilon R1-mediated signaling and effector function by the Syk-selective inhibitor, piceatannol.

In RBL-2H3 rat tumor mast cells, antigens that cross-link the high affinity cell surface IgE receptor, Fc epsilon R1, activate at least two receptor-associated protein-tyrosine kinases, Lyn and Syk, and cause the tyrosine phosphorylation of the receptor beta and gamma subunits, PLC gamma 1, Vav, and other proteins. Cross-linking antigens also induce increased phosphatidylinositol turnover, Ca2+ mobilization, secretion, actin polymerization, spreading, and membrane ruffling. We have used the protein-tyrosine kinase inhibitor, piceatannol (3,4,3',5'-tetrahydroxy-trans-stilbene), to explore the coupling of specific kinases to cellular responses. Piceatannol preferentially inhibits the activity of Syk as compared with Lyn when added to in vitro assays with isolated enzymes. Treatment of RBL-2H3 cells with piceatannol strongly inhibits the antigen-stimulated activation of Syk measured in anti-Syk and anti-phosphotyrosine immune complex kinase assays at concentrations that have no effect on receptor subunit phosphorylation and maintain or increase the activity of Lyn in anti-phosphotyrosine immune complex kinase assays. In cells metabolically labeled with [32P]orthophosphate, piceatannol inhibits the antigen-stimulated tyrosine phosphorylation of Syk and most other proteins. However, receptor subunit phosphorylation is unchanged. Selective inhibition of Syk in this manner blocks receptor-mediated down-stream cellular responses (inositol 1,4,5-trisphosphate production, secretion, ruffling, and spreading) while having only minor effects when these responses are induced with drugs that bypass the receptor complex. These results reveal receptor-mediated Lyn activation as a relatively piceatannol-insensitive event that may contribute to receptor subunit phosphorylation and Syk activation but does not per se elicit cellular responses. Receptor-mediated Syk activation on the other hand is highly sensitive to piceatannol, is essential to Fc epsilon R1-mediated cellular responses, and, based on the increased phosphorylation of Lyn in piceatannol-treated cells, may be involved in terminating Lyn activity.

Phosphorylation of human erythrocyte band 3 by endogenous p72syk.

The anion transporter, band 3, is the major tyrosine kinase substrate in both red cell membranes and intact human erythrocytes. Using antibodies to various protein-tyrosine kinases, we found that p72syk and p56/53lyn are present in human red cells, while p56lck, pp60src, p59fyn, and p55blk are absent. Treatment of intact red cells with a combination of vanadate and hydrogen peroxide dramatically increased the tyrosine phosphorylation of band 3. This treatment increased the tyrosine kinase activity of p72syk and decreased the activity of p56/53lyn in immune complex kinase assays. Band 3 was found to be associated in immune complexes with p72syk but not with p56/53lyn. These findings suggest that p72syk is responsible, at least in part, for the tyrosine phosphorylation of band 3 in human erythrocytes.

p72syk tyrosine kinase is activated by oxidizing conditions that induce lymphocyte tyrosine phosphorylation and Ca2+ signals.

We have used H2O2 as a pharmacologic agent to examine the effects of oxidizing conditions on lymphocyte signal pathways. Treatment of Ramos cells with 5-10 mM H2O2 gave rapid and strong tyrosine phosphorylation of multiple cellular proteins and activation of p72syk to levels equal to or greater than that observed upon surface Ig cross-linking. Strong Ca2+ signals that could be blocked by the tyrosine kinase inhibitor herbimycin A were also observed under these conditions. However, there was no increase in activity for the Src family kinases p56lck, p59fyn, or p56/p53lyn. Our findings that the p72syk tyrosine kinase responds to H2O2 treatment of cells suggest that this kinase is likely to contribute to cellular tyrosine phosphorylation and calcium signaling induced by oxidizing conditions. Furthermore, H2O2 may be useful as a pharmacologic agent to distinguish the effects of p72syk-related kinases from those of Src family kinases.

Cell cycle-specific activation of the PTK72 protein-tyrosine kinase in B lymphocytes.

The engagement of cell-surface antigen receptors on B lymphocytes by anti-IgM antibodies leads to the rapid tyrosine phosphorylation and activation of the protein-tyrosine kinase, PTK72. High concentrations of anti-IgM, which promote cell cycle entry and progression through G1, result in a biphasic change in the state of tyrosine phosphorylation of PTK72. An initial, rapid increase is seen within 5 min, which slowly declines to the level found in resting cells over a period of 9 h. A second increase is then observed 18-30 h following the initial stimulation. Low concentrations of anti-IgM, which promote cell cycle entry but not progression through G1, result only in the initial, rapid phosphorylation. The polyclonal mitogens lipopolysaccharide and dextran sulfate, which stimulate both cell cycle entry and progression to late G1, result only in the second, late phase of tyrosine phosphorylation. This second phase of elevated tyrosine phosphorylation is cell cycle-dependent, as demonstrated by its appearance in cells blocked at G1/S and absence in cells blocked at G2/M. The increase in the tyrosine phosphorylation of PTK72 is accompanied by an increase in its activity with no change in its concentration. These data suggest a possible second role for PTK72 in the commitment of activated B cells to proliferation.