Targeted deletions of complement lectin pathway genes improve outcome in traumatic brain injury, with MASP-2 playing a major role.

The lectin pathway (LP) of complement activation is believed to contribute to brain inflammation. The study aims to identify the key components of the LP contributing to TBI outcome as possible novel pharmacological targets. We compared the long-term neurological deficits and neuropathology of wild-type mice (WT) to that of mice carrying gene deletions of key LP components after experimental TBI. WT or MASP-2 (Masp2-/-), ficolin-A (Fcna-/-), CL-11 (Colec11-/-), MASP-1/3 (Masp1-/-), MBL-C (Mbl2-/-), MBL-A (Mbl1-/-) or MBL-/- (Mbl1-/-/Mbl2-/-) deficient male C57BL/6J mice were used. Mice underwent sham surgery or TBI by controlled cortical impact. The sensorimotor response was evaluated by neuroscore and beam walk tests weekly for 4 weeks. To obtain a comparative analysis of the functional outcome each transgenic line was rated according to a health score calculated on sensorimotor performance. For selected genotypes, brains were harvested 6 weeks after injury for histopathological analysis. MASP-2-/-, MBL-/- and FCN-A-/- mice had better outcome scores compared to WT. Of these, MASP-2-/- mice had the best recovery after TBI, showing reduced sensorimotor deficits (by 33% at 3 weeks and by 36% at 4 weeks). They also showed higher neuronal density in the lesioned cortex with a 31.5% increase compared to WT. Measurement of LP functional activity in plasma from MASP-2-/- mice revealed the absence of LP functional activity using a C4b deposition assay. The LP critically contributes to the post-traumatic inflammatory pathology following TBI with the highest degree of protection achieved through the absence of the LP key enzyme MASP-2, underlining a therapeutic utility of MASP-2 targeting in TBI.

[Development and Evaluation of a Health Promotion Programme for Low-Qualified Workers with Special Attention to Transfer]. / Entwicklung und Evaluation eines Gesundheitsförderungsprogramms für Un- und Angelernte unter besonderer Berücksichtigung der Transfersicherung.

A health promotion programme for low-qualified workers was developed with focus on resource and stress management (ReSuM) and with special attention to transfer. ReSuM is a multiplier concept and combines a team intervention for low-qualified workers with an intervention for their supervisors. The effectiveness and efficiency were successfully evaluated within a comprehensive evaluation strategy.

Consumption of mannan-binding lectin during abdominal aortic aneurysm repair.

OBJECTIVE: Patients undergoing abdominal aortic aneurysm (AAA) repair are exposed to an ischaemia-reperfusion injury (IRI), which is in part mediated by complement activation. We investigated the role of the novel lectin pathway of complement during IRI in patients undergoing AAA repair. METHODS: Patients undergoing elective open infrarenal AAA repair had systemic blood samples taken at induction of anaesthesia, prior to aortic clamping, prior to aortic declamping and at reperfusion. Control patients undergoing major abdominal surgery were also included. Plasma was assayed for levels of mannan-binding lectin (MBL) using ELISA techniques. Consumption of plasma MBL was used as a measure of lectin pathway activation. RESULTS: Twenty-three patients undergoing AAA repair and eight control patients were recruited. No lectin pathway activation could be demonstrated in the control patients. AAA patients experienced a mean decrease in plasma MBL levels of 41% representing significant lectin pathway activation (p = 0.003). CONCLUSION: Consumption of MBL occurs during AAA repair, suggesting an important role for the lectin pathway in IRI. Specific transient inhibition of lectin pathway activity could be of significant therapeutic value in patients undergoing open surgical AAA repair.

Low-level expression of functional foamy virus receptor on hematopoietic progenitor cells.

Foamy viruses have several qualities favorable for vector development: they are not known to cause disease; they can transduce stationary cells; and the foamy virus receptor is expressed on a wide variety of cells. Here, we analyzed the level of virus receptor expression on hematopoietic progenitor cells. Foamy virus binding was measured by a flow cytometric assay and was found to be considerably reduced in hematopoietic progenitors cell lines as well as in primary CD34(+) cells when compared to fibroblasts. Retroviral vectors based on murine leukemia virus (MLV) pseudotyped with a foamy virus envelope transduced hematopoietic cell lines with a more than 10-fold lower efficiency than fibroblasts. Moreover, less than 1% of primary CD34(+) hematopoietic progenitor cells were transduced with the foamy virus pseudotypes, while gene transfer efficiencies of 8-40% were achieved using pseudotypes with amphotropic envelope or the G protein of vesicular stomatitis virus. In conclusion, the expression of functional foamy virus receptors on hematopoietic progenitors cells was found to be insufficient to achieve high levels of gene transfer into CD34(+) hematopoietic progenitor cells with cell-free vector supernatants using current transduction protocols.

A threshold-like measure for the assessment of olfactory sensitivity: the "random" procedure.

Many tests of olfactory dysfunction are either too complex, too expensive, or too time-consuming to be of use in routine clinical testing. Thus, the present multicenter study was undertaken to investigate a new approach, the so-called "random" test. In this test different concentrations of citronellal and phenyl ethyl alcohol are applied according to a pre-established order; patients are asked to identify the odor if possible. The test score is the sum of correctly identified odors. Test administration takes about 10 min. Two studies were performed. Basic characteristics of the test were explored in experiment 1 in 176 healthy subjects (76 male, 100 female; age 12-85 years, mean age 30 years), namely test-retest reliability, correlation with other measures of olfactory sensitivity, and sensitivity of the test to differences in age and gender. In the second experiment the test was tried in 97 patients (45 male, 52 female; age 19-78 years, mean age 47 years) in a clinical environment to investigate its usefulness in diagnosing olfactory loss. The "random"-test was found (1) to exhibit a test-retest reliability similar to that reported for established measures of olfactory function (r = 0.71; P < 0.001), (2) to correlate with other measures of olfactory sensitivity (0.82 > r > 0.60; P < 0.001), (3) to differentiate between expected differences in olfactory sensitivity in relation to gender (t > 2.602, P < 0.011), and (4) to discriminate between different degrees of olfactory loss (F > 36.6, P < 0.001). Based on these data, and the fact that the new test requires little time and is easy to use, this approach can be expected to suit clinical needs.

Lack of superinfection interference in retroviral vector producer cells.

Vesicular stomatitis virus G protein (VSV-G)-pseudotyped retroviral vectors have become more feasible for clinical gene transfer protocols since stable tetracycline (tet)-regulated packaging cell lines have become available. Here, we analyzed superinfection interference in VSV-G-pseudotyped and classic amphotropic packaging cell lines. No superinfection interference was observed in VSV-G-pseudotyped packaging cell lines. Thus, integrated retroviral vector genomes accumulated during culture. Similar results were obtained with the amphotropic packaging cells, but to a lesser degree. In addition, VSV-G packaging cells were susceptible to infection with vector particles devoid of envelope proteins, which are produced by these cells in high titers when VSV-G expression is suppressed by tetracycline. For both packaging systems, superinfection could be blocked by azidothymidine (AZT). With regard to safety, this study suggests that in clinical protocols amphotropic producer clones should be tested for superinfection interference and VSV-G packaging cells should always be cultured in the presence of AZT.

Amphotropic and VSV-G-pseudotyped retroviral vectors transduce human hematopoietic progenitor cells with similar efficiency.

One restriction of retroviral gene transfer into hematopoietic stem cells is the low level of amphotropic virus receptor. In the present study, we examined whether retroviral vectors pseudotyped with the G-protein of vesicular stomatitis virus (VSV) can overcome this restriction. Human progenitor cells purified by magnetic beads and cell sorting were transduced with an amphotropic or VSV-G-pseudotyped retroviral vector containing the truncated human nerve growth factor receptor as a marker gene. Cells were prestimulated with flt-3 ligand, stem cell factor, and interleukin-3 and transduced on fibronectin. Marker gene expression was analyzed by flow cytometry. Transduction efficiencies of amphotropic and VSV-G-pseudotyped virus for CD34+ cells did not differ significantly. Gene transfer into CD34+CD38- cells, which are enriched in more immature progenitors, was not restricted and transfer efficiencies for this subset were also similar for both pseudotypes. The addition of fibronectin improved gene transfer with the amphotropic vector considerably (5- to 19.3-fold, mean 12.6), while the effect on the VSV-G-pseudotype was far less pronounced (1- to 3.9-fold, mean 2.1, P = 0.04). In conclusion, high levels of gene transfer to human hematopoietic progenitors were achieved with an optimized transduction protocol, and transduction efficiencies could not be improved further by the use of VSV-G-pseudotypes.

Metabolic diseases and their possible link to risk indicators of periodontitis.

BACKGROUND: During the last few years, risk assessment has become one of the main topics of periodontal research. Therefore, the aim of this study was to determine whether a predisposition to metabolic disorders such as diabetes mellitus (in the absence of diagnosed diabetic disease) or hyperlipidemia may be risk indicators for periodontitis. METHODS: One hundred patients ranging in age from 40 to 70 years were examined. The patients were classified as having impaired glucose tolerance (IGT) but no manifest diabetes (56 patients), hyperlipidemia (17 patients, HL), or normal metabolic status (27 control patients). Probing depth (PD), attachment level (AL), plaque index (PI), and gingival bleeding on probing (BOP) were recorded. Serum antibody titers (SAT) to A. actinomycetemcomitans (A.a.), P. intermedia (P.i.), and P. gingivalis (P.g.) were determined by enzyme-linked immunosorbent assay (ELISA). Pooled subgingival plaque samples were analyzed using indirect immunofluorescence to detect the same organisms. In addition, respiratory burst activity of peripheral polymorphonuclear leukocytes (PMN) was evaluated by chemiluminescence (CL). RESULTS: No significant differences were observed between the IGT group and normal controls in the following parameters: 1) percentage of sites exhibiting BOP; 2) mean PI; 3) mean PD and AL; 4) percentage of periodontal microorganisms; and 5) increased SAT. The IGT probands exhibited a significantly higher mean serum level of triglycerides, as well as higher formyl-methionyl-leucyl-phenylalanine (FMLP)-stimulated PMN chemiluminescence than the control group. Patients with hyperlipidemia (HL) showed a significantly higher number of sextants with increased PD (73.4%) than the control group (50.6%). Similar results were obtained when comparing the percentage of all sites with increased PD (HL = 16.7%, control 12.3%). The mean FMLP-stimulated CL in patients with hyperlipidemia was significantly higher than the control group. When looking at all patients, there was a small but statistically significant correlation between PD and lipid levels. In addition, a significant correlation was observed between lipid serum levels and the FMLP-stimulated chemiluminescence. CONCLUSIONS: These findings suggest that abnormal glucose tolerance, which is a predisposing factor for diabetes mellitus, does not appear to be a risk indicator for periodontal disease. On the other hand, impaired lipid metabolism does seem to be a risk indicator for periodontitis.

Corticotropin-releasing-hormone lacks analgesic properties: an experimental study in humans, using non-inflammatory pain.

The antinociceptive potency of corticotropin-releasing-hormone (CRH) has been established in several animal studies in which both central and peripheral sites of action were considered. However, there have not yet been any experimental trials, besides one attempt using clinical dental pain demonstrating the potential analgesic properties of CRH in humans. For this reason, we studied the effect of CRH on experimental heat pain sensitivity in 18 healthy men, using a double-blind, cross-over and placebo-controlled design. A dose of 100 microg (i.v.) was chosen because of its well-known neuroendocrine effects in humans. The pain parameters assessed were, visual analog scale (VAS) ratings for pain intensity and pain unpleasantness, pain thresholds and scores for discrimination ability. To differentiate between a direct analgesic effect of CRH and indirect effects via evoked hormonal responses in the hypothalamic-pituitary-adrenocortical (HPA) system (beta-endorphin, ACTH, cortisol), CRH was applied with and without a pre-treatment with dexamethasone. In neither of the two conditions was there any systematic change in our pain parameters. This failure to find any evidence suggesting an analgesic action of CRH or of the subsequent hormones of the HPA system was obtained despite the fact that CRH produced clear neuroendocrine responses such as increases in the plasma concentration of beta-endorphin and cortisol. It is unclear whether the lack of analgesic action of CRH is due to its non-existence in humans, due to the use of a pain model which does not assess minute changes in pain sensitivity and does not trigger substantial inflammatory responses, or due to an insufficient dose of CRH.

Relation of olfactory event-related potentials to changes in stimulus concentration.

OBJECTIVE: The aim of this study was to investigate the influence of odorant concentration on the olfactory event-related potential (OERP). METHODS: OERP were evaluated in 8 men and 8 women (17-34 years of age) in response to 4 concentrations of vanillin (7, 28, 56 and 84% v/v). Sixteen presentations of each concentration (stimulus duration 200 ms, interval 40 s, flow 81/min) were applied in a randomized order. EEG recordings were made at 3 midline sites (pos. Fz, Cz, Pz). Amplitudes and latencies of four peaks were measured (latencies in ms at Pz after stimulation with 84% v/v vanillin): P1 (277), N1 (348), P2 (412) and P3 (496). Statistical analysis was performed with MANOVAs ('concentration', 'recording site' = within-subject-factors; 'age' as covariate). RESULTS: With increasing stimulus concentration amplitudes became significantly larger; this was most pronounced for P3 (P1N1: F = 2.90, P < 0.05; N1P2: F = 5.15, P < 0.01; N1P3: F = 35.7, P < 0.001; P3: F = 38.6; P < 0.001). Correspondingly, latencies shortened with increasing concentrations (P1: F = 25.2; N1: 17.51; P2: 14.8; P3: 13.4; all P < 0.001). While there was no correlation between OERP amplitudes and butanol odor detection thresholds, latencies were the shorter the lower the subjects' thresholds (coefficients of correlations for peak latencies at Cz for 84% v/v: P1 rl5 = -0.59; N1 rl5 = 0.58; P2 r15 = -0.55; P3 r15 = -0.45). CONCLUSIONS: The results indicated that both OERP amplitudes and latencies are related to the concentration of olfactory stimuli. They also suggested that latencies exhibit a stronger relation to changes in stimulus intensity compared to OERP amplitudes.

"Sniffin' sticks": screening of olfactory performance.

"Sniffin' Sticks" is a new test of nasal chemosensory performance based on pen-like odour-dispensing devices. This portable test is suited for repetitive, inexpensive screening of odour identification. The test includes a forced odour-identification task for seven odours performed by means of a list of four items (multiple-choice). In 146 subjects the basic screening test was compared to a down-scaled version of the UPSIT (CC-SIT). Sniffin' Sticks exhibited a relatively higher coefficient of correlation with the subjects' age; they also demonstrated the women's superior olfactory sensitivity more pronounced when compared to men. In addition, the coefficient of correlation between age and olfactory performance was slightly higher when the sticks were used. Preliminary investigations in nine patients with impaired olfactory function (i.e., anosmic or hyposmic patients) revealed significantly lower scores in patients compared to healthy controls matched for age and sex (p < 0.001). It is concluded that Sniffin' Sticks may be useful in the routine clinical assessment of olfactory performance where both time and costs matter.

Intranasal chemoreception in patients with multiple chemical sensitivities: a double-blind investigation.

Multiple chemical sensitivities (MCS) has become an increasingly frequent diagnosis assigned to patients with symptoms associated with exposures to environmental chemicals. Since the characteristic symptoms of MCS are triggered by very low concentrations of chemicals, in the range of olfactory thresholds, it is widely believed that the intranasal chemoreceptive senses are involved in the pathophysiology of MCS. Thus, the present study addressed both the olfactory and trigeminal systems: using a double-blind approach we investigated whether MCS patients show differences in responses after exposure to either room air or low concentrations of a widely used chemical agent (2-propanol). A total of 23 patients participated in the experiments (mean age 47 years; 13 female, 10 male). MCS was diagnosed according to Cullen's criteria Performance of the nasal chemical senses was established by means of chemosensory event-related potentials (CSERP) and subjective measures of olfactory function (odor discrimination, phenylethyl alcohol odor thresholds). CSERP were recorded in response to olfactory (H2S), and trigeminal (CO2) stimuli. The study provided three major results: (1) Approximately 20% of patients diagnosed with MCS presented symptoms regardless of the type of challenge, suggesting the susceptibility of MCS patients to unspecific experimental manipulations. (2) Changes in CSERP latencies indicated a change in the processing of both olfactory and trigeminal stimuli. (3) While odor threshold remained unchanged, the patients' ability to discriminate odors decreased after exposure to room air. In contrast, this decrease was less pronounced after exposure to 2-prop. Summarily, MCS patients respond to challenge with 2-prop with changes of chemosensory perception which might increase their susceptibility to environmentally volatile chemicals. Changes in the pattern of event-related potentials are interpreted as the possible change of the orientation of cortical generators, i.e., neuronal populations that were involved in the processing of chemosensory information. However, investigations in healthy controls are needed in order to draw further conclusions.

Loss of olfactory function leads to a decrease of trigeminal sensitivity.

Healthy controls were compared to patients with decreased olfactory sensitivity (n = 32) to investigate interactions between the olfactory and trigeminal systems. Amplitudes of chemo-somatosensory event-related potentials in response to suprathreshold trigeminal stimuli (CO2) were found to be smaller in patients (P < 0.05) indicating a decrease of trigeminally mediated sensations.

Assessment of analgesia in man: tramadol controlled release formula vs. tramadol standard formulation.

OBJECTIVE: The present study tested analgesia produced by a new controlled release formulation of tramadol. The investigation employed an experimental pain model based on chemo-somatosensory event-related potentials (CSSERP) in response to painful chemical stimuli applied to the nasal mucosa. STUDY: Twenty healthy volunteers participated in the experiments, which followed a controlled, randomised, double-blind, 3-way cross-over design. Each of the three medications (tramadol 100 mg [T100], tramadol controlled release 100 mg [TCR100] and tramadol controlled release 150 mg [TCR150]) was administered orally to fasting subjects. There was at least a 6 day washout period between tests. Each experiment was divided into five sessions, which took place before and 2, 4, 6, and 12 h after drug administration. In addition to the assessment of CSSERP, subjects rated the intensity of both the tonic and phasic painful stimuli. Nonspecific drug effects were also monitored by means of frequency analysis of the spontaneous EEG, ratings of adverse effects, and the subjects' performance in a tracking task. RESULTS: The significant reduction of amplitude N1 at central recording positions indicated that TCR 150 was the most effective analgesic 12 h after administration. Both 6 and 12 h after administration TCR 100 was more effective in terms of analgesia compared to T100. In addition, TCR100 appeared to produce fewer adverse effects than the standard formulation of tramadol. CONCLUSIONS: The controlled release formulation can be expected to become a valuable tool in peroral therapeutic regimens for chronic pain.

Tonic pain evoked by pulsating heat: temporal summation mechanisms and perceptual qualities.

The properties of a newly developed tonic heat pain model (THPM), which makes use of pulsating contact heat, were investigated in 18 young men. The most important feature of this model is that repetitive heat pulses with an intensity of 1 degree C above the individual pain threshold are employed. This approach was used to tailor the tonic pain stimulation to the individual pain sensitivity. In the first of two experiments, the effects of pulse frequencies ranging from 5 to 30 pulses per minute (ppm) on ratings of pain intensity and pain unpleasantness (visual analogue scales) were examined. At all frequencies, both ratings increased steadily over the 5-min test period. Frequencies of 15 ppm or more appeared to enhance pain intensity throughout the test period compared to the lower frequencies, but did not appear to alter pain unpleasantness. This suggests that only pain intensity is influenced by slow temporal summation and that a sort of frequency threshold exists for this kind of summation. In the second experiment, the THPM was compared to a well-established form of tonic pain stimulation, the cold-pressor test (CPT); visual analogue scales were again used, and in addition the McGill Pain Questionnaire was employed. The CPT appeared to produce stronger tonic pain than the THPM. However, as is typical with tonic pain, both tonic pain models induced relatively higher values on the affective pain dimension than on the sensory pain dimension. The time course of pain was dynamic in the CPT, with an increase followed by a plateau phase, at least in those subjects who could tolerate the CPT for more than 60 sec. In contrast, as in the first experiment, the pain ratings in the THPM were characterized by a slow and steady increase over time. Moreover, there was absolutely no indication of a dichotomy between "pain-sensitive" and "pain-tolerant" individuals in the THPM, although such a dichotomy was evident in the CPT. This implies that the distinction between pain-sensitive and pain-tolerant individuals can be made only with the CPT, and that this distinction represents individual differences in peripheral vascular reactions to cold rather than in pain perception. In conclusion, the THPM appears to produce a stable and predictable temporal pattern of tonic pain with a predominant affective component, and to be suitable for application in the majority of individuals without causing undue discomfort.

Pain perception in depression: relationships to symptomatology and naloxone-sensitive mechanisms.

A decrease in pain sensitivity during acute depression has been observed in several studies, apparently related to the severity of symptomatology. However, the question remains whether this relationship can be found only in heterogeneous groups of depressive patients or also in a single diagnostic group, such as major depression. In the present study, pain thresholds were assessed in 20 patients with major depression (DSM-III-R) and in 20 healthy controls. Two threshold methods with a differing impact of reaction time on the results were used. Contact heat was applied as a natural source of pain. With both methods the pain thresholds were significantly increased in the depressive patients. No relationship was found to the various symptoms of depression assessed by psychopathometric scales. In contrast to the pain thresholds, the thresholds of skin sensitivity for nonnoxious stimuli (warmth, cold, vibration) were only slightly increased. In subsamples (N = 10 in each group), naloxone (5 mg i.v.) and placebo were administered in a double-blind design. No systematic changes in pain thresholds occurred under either treatment. Our findings suggest that the decrease in skin sensitivity in major depression is specific to pain and not due to an increased reaction time. Moreover, the decrease appears to be related neither to a naloxone-sensitive mechanism nor to symptomatology.

Pain sensitivity in recovered anorexics, restrained and unrestrained eaters.

The heat pain threshold and local skin temperature were assessed in 23 former anorexic in-patients with an 'intermediate' (N = 9) or 'good' outcome (N = 14) and in 21 restrained and 20 unrestrained eaters. All subjects were female. The group means of the pain thresholds did not differ significantly from each other, suggesting that the homogeneous increase in pain thresholds we had previously observed in acutely ill eating disorder patients is state dependent. However, a sizeable percentage of the restrained eaters (29%) had pain thresholds clearly above the normal range. Thus it may well be that restrained eating carries a risk of reducing pain sensitivity. Pain threshold and skin temperature correlated significantly (r = -0.63) only in the group of patients with an intermediate outcome, a finding resembling that obtained in acute anorexics. This suggests that peripheral thermoregulation and pain sensitivity are linked in the acute and moderately improved phases of anorexia nervosa.

Theoretical and empirical considerations on the relation between 'body image', body scheme and somatosensation.

The authors first discuss possible interactions between the hitherto neglected neurophysiological and neuropsychological factors and the traditionally accepted cognitive and affective factors in 'body image' formation. They then report on a study of the relation between body size perception (video distortion technique, image marking procedure, kinaesthetic size estimation apparatus) and somatosensation (thermal, pain and vibration thresholds) in young women. Included in the study were questionnaires on eating behaviour and motivation, body attitude or body satisfaction, and depressive mood and thoughts. Neither the somatosensory nor the questionnaire variables explained the difference between 'overestimators' and 'underestimators' of body size. However, these variables did explain the difference between 'good perceivers' and 'poor perceivers' (degree of deviation from actual body size) in the video distortion technique, with a somewhat larger contribution by the somatosensory variables. The latter finding, although clearly preliminary, should stimulate further investigations of the relationship between somatosensory variables and body size perception.