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.

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.

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.

Synthesis and protein-tyrosine kinase inhibitory activities of flavonoid analogues.

Treatment of o-hydroxyacetophenones 2a-e with excess lithium bis(trimethylsilyl)amide followed by dialkyl carbonates gave alkyl 3-(2-hydroxyaryl)-3-oxopropanoates 3a-e. The latter substances were transformed through the reaction of their magnesium chelates with benzoyl chlorides into a series of 3-(alkoxycarbonyl)-2-arylflavones, which were subsequently elaborated into a variety of flavonoids. These compounds were tested for their abilities to inhibit the in vitro protein-tyrosine kinase activity of p56lck, an enzyme which is thought to play a key role in mediating signal transduction from the CD4 receptor during lymphocyte activation. All of the active compounds had either an amino or a hydroxyl substituent at the 4'-position of the 2-aryl ring. The most active substance prepared in this study is compound 17c, which is approximately 1 order of magnitude more potent than the natural product quercetin (1). Compound 17c was a competitive inhibitor of p56lck with respect to ATP and was highly selective for the inhibition of protein-tyrosine over protein-serine/threonine kinases.

An alternative method of panning for rat B lymphocytes.

A new method of panning for B lymphocytes is described in which the ability of the sIg+ cells to adhere depends on the nature and concentration of nonspecific protein used rather than on the use of anti-immunoglobulin. Rat lymph node cells were suspended in 3% bovine serum albumin in Tris-buffered Hanks' and incubated in tissue culture flasks to allow adherence to the plastic. The recovered bed of adherent cells was shown by flow cytometry to be greater than 90% surface immunoglobulin positive and MHC class II positive while containing very few T cells. This adherent fraction was subsequently treated with anti-T cell antibody plus baby rabbit complement to produce a highly purified sIg+ cell population containing no detectable T cells. The sIg+ cells obtained by this panning procedure were functionally active in BCGF and BCDF assays. This method provides an easy and inexpensive alternative to conventional panning with anti-immunoglobulin and also eliminates the possibility of B cell activation by exposure to anti-immunoglobulin-coated surfaces.

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.

[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.

[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.

[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.

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.

[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.

Polyclonal activation of primed rat B cells.

In recent years, murine and human virgin B lymphocytes have been used to examine the steps necessary for polyclonal activation. In these models mitogens are used in conjunction with lymphokines to determine which signals are responsible for regulating B-cell triggering, proliferation, and differentiation. While progress has been made in understanding these events as they occur in virgin B cells, very little evidence exists to suggest whether these models of activation also apply to the memory B-cell population. In this report we have described an antigen-specific, secondary in vitro immune response using cells isolated from lymph nodes draining the site of antigen injection. Unfractionated cells, B cells, and size-fractionated cells from dinitrophenyl-keyhole limpet hemocyanin (DNP-KLH)-primed rats were challenged in vitro with DNP-KLH, lipopolysaccharide plus dextran sulfate (LPS/DxS), and T-cell factors. We have consistently found, under all these conditions, that antigen challenge of primed cells results in the production of DNP-specific IgG antibody while stimulation with LPS/DxS plus T-cell factors results only in the polyclonal activation of virgin B cells; no antigen-specific IgG secretion is seen. This suggests that acquisition of memory status is associated with a loss in responsiveness to LPS/DxS-induced differentiation.