Expression of PD-1 and Tim-3 is increased in skin of patients with bullous pemphigoid and pemphigus vulgaris.

BACKGROUND: Bullous pemphigoid (BP) and pemphigus vulgaris (PV) are common autoimmune bullous dermatoses (AIBD) characterized by blisters and erosions. Treatment options are limited and often insufficient. Immune checkpoint receptors play critical roles in immune homoeostasis and self- tolerance. Targeting checkpoint receptors is highly efficient in treatment of various cancers, but often also associated with autoimmune side effects. OBJECTIVES: We therefore aimed to investigate the expression of immune checkpoint receptors in patients with BP and PV. METHODS: We analysed expression of the checkpoint receptors programmed cell death protein 1 (PD-1), T-cell immunoglobulin and mucin domain 3 (Tim-3) and lymphocyte activation gene 3 (Lag-3) in lesional skin of patients with BP and PV compared to healthy control skin as well as the expression patterns of PD-1 and Tim-3 on various infiltrating immune cells in skin sections of AIBD by immunohistochemistry and immunofluorescence. We also measured serum levels of soluble PD-1, Tim-3 and Lag-3 in AIBD patients by ELISA. RESULTS: We report on increased expression of PD-1 and Tim-3, but not Lag-3, in lesional skin of patients with BP and PV. Investigating the expression pattern of PD-1 and Tim-3 on different cutaneous immune cells, we observed significant upregulation of PD-1 predominantly on infiltrating CD8 T cells and upregulation of Tim-3 on CD8 T cells as well as macrophages. CONCLUSIONS: Our results suggest exploring immune checkpoint receptors as novel therapeutic targets using an agonistic approach in autoimmune bullous diseases.

The association between collagen and bone biomarkers and radiographic osteoarthritis in the distal tarsal joints of horses.

BACKGROUND: Osteoarthritis (OA) of the distal intertarsal (DIT) and tarsometatarsal (TMT) joints occurs commonly. Synovial fluid (SF) biomarkers of collagen and bone turnover have potential clinical value. OBJECTIVES: To measure SF biomarker concentrations from DIT and TMT joints in adult horses and determine if they correlate with radiographic OA severity and are higher in joints with radiographic OA compared to controls. STUDY DESIGN: Cross-sectional. METHODS: Radiographic OA of DIT and TMT joints was evaluated from adult horses (5-35 years old). Overall radiographic scores divided horses into those with mild or moderate radiographic OA (16 joints from 9 horses) or controls (13 joints from 9 horses). Direct biomarkers of OA (Carboxypropeptide of type II collagen = CPII, carboxy-neoepitope of type II collagen exposed after collagenase-cleavage = C2C, Bone alkaline phosphatase = BAP and Chondroitin sulfate epitope = CS846) were measured via ELISA and CPII/C2C was calculated. Biomarkers were correlated with radiographic findings and concentrations from those with radiographic OA to control joints and were compared. RESULTS: Concentrations of CPII (R = 0.84, P<0.001), C2C (R = 0.69, P<0.001) and BAP (R = 0.41, P = 0.03) as well as CPII/C2C (R = 0.69, P<0.001) values positively correlated with overall radiographic scores. Adjusted means ± s.d., after controlling for age, for CPII (P<0.001), C2C (P<0.001), CPII/C2C (P = 0.004) and BAP (P = 0.05) were significantly higher in DIT and TMT joints with radiographic OA (CPII: 2174.45 ± 1064.01; C2C: 233.52 ± 51.187; CPII/C2C: 9.01 ± 4.09; BAP: 21.98 ± 15.34) compared to controls (CPII: 594.53 ± 463.05; C2C: 153.12 ± 48.95; CPII/C2C: 3.96 ± 2.38; BAP: 12.76 ± 3.61). CPII (P<0.001), C2C (P = 0.001) and CPII/C2C (P = 0.001) were significantly higher with moderate radiographic OA (CPII: 2444.61 ± 772.78; C2C: 248.90 ± 44.94; CPII/C2C: 9.47 ± 2.97) compared to controls (CPII: 658.38 ± 417.36; C2C: 156.49 ± 47.61; CPII/C2C: 4.15 ± 2.04), with CPII also showing significantly higher concentrations (P = 0.04) with mild radiographic OA compared (1515.00 ± 584.95) to controls (658.38 ± 417.36). There were no differences in CS846 concentrations between radiographic OA and control joints. Age positively correlated with CPII (R = 0.48, P = 0.01) and C2C (R = 0.44, P = 0.02) concentrations. MAIN LIMITATIONS: Radiographic OA was assessed, not clinical lameness. Controls were not age-matched to those with spontaneous radiographic OA. CONCLUSIONS: There is an association between collagen (CPII, C2C and CPII/C2C) and bone (BAP) biomarkers and radiographic OA in the distal tarsal joints of horses.

Growth and Product Formation of Clostridium ljungdahlii in Presence of Cyanide.

Cyanide is a minor constituent of crude syngas whose content depends on the feedstock and gasification procedure. It is a known poison to metal catalysts and inhibits iron-containing enzymes like carbon monoxide dehydrogenase of acetogenic organisms. Therefore, it is considered a component that has to be removed from the gas stream prior to use in chemical synthesis or syngas fermentation. We show that the growth rate and maximum biomass concentration of Clostridium ljungdahlii are unaffected by cyanide at concentrations of up to 1.0 mM with fructose as a carbon source and up to 0.1 mM with syngas as a carbon source. After the culture is adapted to cyanide it shows no growth inhibition. While the difference in growth is an increasing lag-phase with increasing cyanide concentrations, the product spectrum shifts from 97% acetic acid and 3% ethanol at 0 mM cyanide to 20% acetic acid and 80% ethanol at 1.0 mM cyanide for cultures growing on (fructose) and 80% acetic acid and 20% ethanol at 0.1 mM cyanide (syngas).

Circumferential hoof clamp method of lameness induction in the horse.

A circumferential hoof clamp method to induce controlled and reversible lameness in the forelimbs of eight horses was assessed. Peak vertical forces and vertical impulses were recorded using a force plate to verify induced lameness. Video recordings were used by blinded observers to determine subjective lameness using a 0-5 scale and any residual lameness following clamp loosening. Tightening of clamps resulted in consistent, visible lameness in the selected limbs in all horses. Lameness was confirmed by significant decreases from baseline in the peak vertical force (P <0.01). Lameness was also confirmed subjectively by elevated median scores (0 at baseline and 2 during lameness). Lameness was not immediately reversible after clamp loosening (median score 1.5), but horses were not obviously lame after clamp removal and were no different from initial baseline (median score 0.5) approximately 3 days later.

Evaluation of the Essen Rotary as a new technique for bacterial swabs: results of a prospective controlled clinical investigation in 50 patients with chronic leg ulcers.

Most chronic wounds are colonised with different microorganisms, especially problematic bacteria like methicillin-resistant Staphylococcus aureus (MRSA), which represent an increasing therapeutic challenge in the modern wound therapy regimen. Therefore, it is essential to specify the bacteria in wounds for an individual-specific treatment. In most patients, an exemplary bacterial swab is taken from the centre of the wound surface. This so-called Levine technique is propagated currently as the gold standard. The aim of our clinical investigation was to compare the results of different swab techniques to the new established Essen Rotary. In this monocentric prospective investigation, 50 patients with chronic leg ulcers were examined consecutively. The results of our clinical study show that bacteria are heterogeneously spread on wound surfaces. The analysis of the semiquantitative measured results showed that the Essen Rotary could detect significant more bacteria with a total amount of 111 bacteria (P = 0·049) compared to usual swab techniques. Considerably, only the Essen Rotary identified five compared to three MRSA-patients detected by other techniques. The Essen Rotary is an efficient, economic and uncomplicated modification of bacteriological swab techniques which detects significant more bacteria compared to other conventional swab techniques. Therefore, the Essen Rotary may become the new gold standard in routinely taken bacteriological swabs especially for MRSA screenings in patients with chronic leg ulcers.

Clinical assessment and outcome of a single-layer technique for anastomosis of the small intestine in horses.

In order to assess postoperative outcome in horses undergoing end-to-end anastomosis of the small intestine, performed using a one-layer technique, 15 horses that underwent exploratory coeliotomy, resection of the small intestine and end-to-end anastomosis using a continuous Lembert pattern were studied. Information on the age, breed, sex, diagnosis, treatment, complications and outcome of each case were obtained from medical records. Follow-up information was obtained via telephone conversations with clients and trainers. Five of the horses had short-term postoperative complications: one had postoperative ileus (POI), colic and peritonitis, one had POI and colic, two had POI only and one had diarrhoea only. A second exploratory coeliotomy was recommended in two of the 15 horses (13 per cent). The short-term survival rate, defined as survival up to the time of discharge from the hospital, was 93.3 per cent (14 of 15 horses). The long-term survival rate, defined as survival for at least 12 months after the surgery, was 84.6 per cent (11 of 13 horses followed up).

[Carotid artery stenting for acute stroke]. / Karotisstent beim akuten Schlaganfall.

For patients with acute ischemic stroke due to total occlusion of the internal carotid artery (ICA), an effective intervention to improve neurologic symptoms and clinical outcome has not yet been established. Some authors have reported successful revascularization for patients with acute stroke symptoms secondary to ICA occlusion only in isolated series and case reports. Emergency recanalization and carotid artery stent placement can improve neurologic outcome in selected patients with acute ischemic stroke and total occlusion of the ICA.

Relationship of apolipoprotein E and age at onset to Parkinson disease neuropathology.

Previous studies investigating the association between apolipoprotein E (APOE) genotypes and Parkinson disease (PD) have yielded conflicting results, and only a few have addressed APOE as a possible determinant of PD pathology. Therefore, we aimed to evaluate the relationship between APOE and PD as well as APOE and PD pathology. We studied 108 pathologically verified patients with PD and 108 controls pair-matched for age and gender. Allele frequencies of APOE differed between patients with PD and controls (p = 0.02). The frequency of epsilon4 allele increased (p = 0.01), whereas that of epsilon3 allele decreased with advancing PD pathology (p = 0.002). Only age of PD onset was an independent predictor for the rate of progression of PD pathology in which late-onset patients appeared to reach end point PD pathology more rapidly than early-onset patients (p = 0.001). In conclusion, our findings suggest that APOE may express its effect on the risk of PD by modifying the occurrence of PD pathology, but age of PD onset seems to be the principal determinant of the progression rate of PD pathology.

Two cases of autosomal recessive generalized dystonia in childhood: 5 year follow-up and bilateral globus pallidus stimulation results.

We report two brothers with an unknown form of early-onset familiar dystonia. Characteristic clinical features are (1) childhood-onset; (2) extrapyramidal motor symptoms; (3) dysarthria; and (4) mental retardation. Additional findings include loss of D(2)-receptors in both basal ganglia and hypoplasia of the cerebellar vermis with dilatation of the fourth ventricle and cisterna magna. There seems to be a progressive and non-progressive form of this clinical entity. Dystonic symptoms of the progressive form that occurred in one of the brothers were alleviated dramatically by bilateral internal globus pallidus (Gpi) stimulation, and the improvement has lasted now for 5 years.

Primary angioplasty: preprocedural pharmacological therapy.

Primary coronary angioplasty has been shown to be an effective reperfusion therapy for patients with acute myocardial infarction, not only for those who present to PTCA centres but also for patients who present to hospitals without angioplasty facilities. With the increasing use of primary angioplasty more patients will be transferred to a (tertiary) PTCA centre. An increase in treatment delay is associated with a worse clinical outcome. The importance of an open infarct-related vessel at acute angiography is becoming clear. Pharmacological pretreatment of patients during transportation to a PTCA centre with the aim to open the infarct-related vessel in advance might be beneficial. Glycoprotein IIb/IIIa receptor blockers seem to be the agents of choice for facilitated PTCA. The safety and (cost) effectiveness of this pretreatment of patients transported to undergo primary angioplasty remain to be evaluated.

Absence of heparan sulfate proteoglycans in Lewy bodies and Lewy neurites in Parkinson's disease brains.

alpha-Synuclein is the major constituent of Lewy bodies and Lewy neurites in Parkinson's disease (PD) and dementia with Lewy bodies (DLB). Relatively little is known about the exact mechanism of alpha-synuclein deposition and fibrillization in these alpha-synucleinopathies. In order to better understand the pathogenesis of alpha-synucleinopathies it is important to identify molecules that regulate the fibrillization of alpha-synuclein. Since it has been demonstrated that heparan sulfate proteoglycans (HSPGs) and glycosaminoglycans (GAGs) promote the conversion of non-fibrillar amyloid beta-protein (Abeta) into neurotoxic fibrillar Abeta in Alzheimer's disease, they might also be involved in alpha-synuclein aggregation. It was the aim of our study to examine the distribution pattern of these macromolecules in PD brains and the possible association with Lewy bodies and Lewy neurites. Although HSPGs clearly colocalized with senile plaques, we were unable to identify HSPGs or GAGs in Lewy bodies and Lewy neurites and therefore concluded that it is likely that alpha-synuclein fibrillization and stabilization occurs independently of the presence of HSPGs or GAGs.

Elevated concentration of cerebrospinal fluid tissue transglutaminase in Parkinson's disease indicating apoptosis.

Tissue transglutaminase (tTG) is activated during the apoptotic cell death cascade and plays a key role in the formation of apoptotic bodies. We found significant elevation of tTG concentration in the cerebrospinal fluid (CSF) of 54 patients with Parkinson's disease (PD) compared with that measured in 34 control subjects, thus showing increased neural cell apoptosis in patients with PD.

Familial cortical tremor with epilepsy and cerebellar pathological findings.

The clinical and neuropathological findings in a patient with familial cortical tremor with epilepsy (FCTE) are described. Clinically, the patient showed cortical myoclonus, tremor, and generalized seizures. Pathological investigation showed cerebellar degeneration and somal sprouting and loss of dendritic tree in Purkinje cells. Striking similarities were found in diseases caused by channelopathies such as spinocerebellar ataxia subtype 6.

Prehospital triage for angiography-guided therapy for acute myocardial infarction.

BACKGROUND: Time between symptom onset and effective reperfusion is of paramount importance in patients with acute myocardial infarction (MI) treated with reperfusion therapy. In the PHIAT (Pre-Hospital Infarction Angioplasty Triage) project, safety and feasibility of in-ambulance electrocardiography facilities for prehospital triage for direct transfer to an interventional centre to undergo immediate coronary angiography and angiography-guided therapy were evaluated. METHODS AND RESULTS: The ambulances were equipped with a defibrillator and electrocardiography unit with computerised electrocardiographic analysis. Patients with acute MI symptoms and fulfilling certain criteria compatible with a large MI were included and pretreated with heparin and aspirin during transportation. During the study period, 284 patients were included. Eleven percent did not have an acute MI. PCI, performed in 94% (n=239) of the patients, was successful in 94%. Prehospital triage reduced time to treatment. In 32% of the patients triage resulted in direct transportation to the interventional centre instead of to the nearest community hospital. All-cause mortality was 9% after a mean follow-up of nine months. No serious bleeding complications were seen. CONCLUSION: Prehospital triage in the ambulance is safe and feasible. A striking percentage (11%) of the identified patients does not have an acute MI and this is more than has been reported from prehospital thrombolysis trials.

Disease-specific accumulation of mutant ubiquitin as a marker for proteasomal dysfunction in the brain.

Molecular misreading of the ubiquitin-B (UBB) gene results in a dinucleotide deletion in UBB mRNA. The resulting mutant protein, UBB+1, accumulates in the neuropathological hallmarks of Alzheimer disease. In vitro, UBB+1 inhibits proteasomal proteolysis, although it is also an ubiquitin fusion degradation substrate for the proteasome. Using the ligase chain reaction to detect dinucleotide deletions, we report here that UBB+1 transcripts are present in each neurodegenerative disease studied (tauo- and synucleinopathies) and even in control brain samples. In contrast to UBB+1 transcripts, UBB+1 protein accumulation in the ubiquitin-containing neuropathological hallmarks is restricted to the tauopathies such as Pick disease, frontotemporal dementia, progressive supranuclear palsy, and argyrophilic grain disease. Remarkably, UBB+1 protein is not detected in the major forms of synucleinopathies (Lewy body disease and multiple system atrophy). The neurologically intact brain can cope with UBB+1 as lentivirally delivered UBB+1 protein is rapidly degraded in rat hippocampus, whereas the K29,48R mutant of UBB+1, which is not ubiquitinated, is abundantly expressed. The finding that UBB+1 protein only accumulates in tauopathies thus implies that the ubiquitin-proteasome system is impaired specifically in this group of neurodegenerative diseases and not in synucleinopathies and that the presence of UBB+1 protein reports proteasomal dysfunction in the brain.

Staging of brain pathology related to sporadic Parkinson's disease.

Sporadic Parkinson's disease involves multiple neuronal systems and results from changes developing in a few susceptible types of nerve cells. Essential for neuropathological diagnosis are alpha-synuclein-immunopositive Lewy neurites and Lewy bodies. The pathological process targets specific induction sites: lesions initially occur in the dorsal motor nucleus of the glossopharyngeal and vagal nerves and anterior olfactory nucleus. Thereafter, less vulnerable nuclear grays and cortical areas gradually become affected. The disease process in the brain stem pursues an ascending course with little interindividual variation. The pathology in the anterior olfactory nucleus makes fewer incursions into related areas than that developing in the brain stem. Cortical involvement ensues, beginning with the anteromedial temporal mesocortex. From there, the neocortex succumbs, commencing with high order sensory association and prefrontal areas. First order sensory association/premotor areas and primary sensory/motor fields then follow suit. This study traces the course of the pathology in incidental and symptomatic Parkinson cases proposing a staging procedure based upon the readily recognizable topographical extent of the lesions.

The importance of patency of the infarct-related artery in treatment of patients with acute myocardial infarction.

OBJECTIVE: To study the impact of patency of the infarct-related artery on the coronary angiogram, both before and after primary angioplasty for acute myocardial infarction, on 30-day mortality. METHOD: Data of 1702 consecutive patients treated with primary angioplasty for acute myocardial infarction were collected prospectively from 1994 to 2000. RESULTS: Patients with a (partially) patent infarct artery before primary angioplasty had less damage to the myocardium and a lower 30-day mortality (1.6% versus 3.4%, p=0.04) compared with patients with an occluded artery. Patients with pre-hospital treatment with aspirin and heparin more often presented with a patent artery before angioplasty (31% versus 20%, p<0.001). After primary angioplasty, 95% of patients had a patent artery with a 30-day mortality of 2.2%. The 5% of patients with failed angioplasty had extensive myocardial damage and a 30-day mortality rate of 17%. CONCLUSION: Patency of the infarct-related artery on the coronary angiogram, both before and after primary angioplasty, has a major impact on 30-day mortality.

Argyrophilic grain disease and Alzheimer's disease are distinguished by their different distribution of tau protein isoforms.

Prominent neuronal and glial tau filamentous inclusions are hallmarks of neurodegenerative tauopathies, among them Alzheimer's disease (AD), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), Pick's disease (PiD), and argyrophilic grain disease (AgD). AgD is a late onset dementia in which pathologically aggregated tau proteins are found in limbic structures in the shape of distinct argyrophilic grains and coiled bodies. Until now tau protein deposits in AgD have not been assessed biochemically. We therefore decided to investigate the electrophoretic profile of pathological tau protein as well as the tau protein isoform composition of filamentous inclusions in AgD cases. A distinct pathological tau doublet at 64 and 69 kDa and a minor 74-kDa band was obtained in two AgD cases with only very mild concomitant AD pathology (Braak stage I), while in two AgD cases with moderate AD pathology (Braak stage II and III, respectively), an additional minor band at 60 kDa was detected. Thus, the pathological tau profile (PTP) in pure AgD cases differs from both the PTPs in AD (tau triplet at 60, 64 and 69 kDa, minor band at 74 kDa) and PiD (major tau doublet at 60 and 64 kDa, minor band at 69 kDa) but not from those in PSP and CBD. Using a two-dimensional gel electrophoresis approach anti-exon 10 antiserum strongly stained the AgD doublet and the minor 74-kDa band, while anti-exon 2 and 3 antisera only faintly stained the 69- and the minor 74-kDa component, thus suggesting that pathological tau aggregates in AgD are mainly made of four-repeat (4R) tau isoforms. Furthermore, in contrast to earlier immunohistochemical studies, we now show biochemically that Ser262 indeed is phosphorylated in the PTP of AgD. Finally, expression of normal tau protein was not found to be altered in AgD. Altogether, our results demonstrate that AgD is characterized by a major tau doublet that is distinct from AD and PiD. AgD, however, shares the pathological tau doublet (64 and 69 kDa) as well as the predominance of 4R tau isoforms with CBD and PSP.

Four related proteins of the Trypanosoma brucei RNA editing complex.

RNA editing in kinetoplastid mitochondria occurs by a series of enzymatic steps that is catalyzed by a macromolecular complex. Four novel proteins and their corresponding genes were identified by mass spectrometric analysis of purified editing complexes from Trypanosoma brucei. These four proteins, TbMP81, TbMP63, TbMP42, and TbMP18, contain conserved sequences to various degrees. All four proteins have sequence similarity in the C terminus; TbMP18 has considerable sequence similarity to the C-terminal region of TbMP42, and TbMP81, TbMP63, and TbMP42 contain zinc finger motif(s). Monoclonal antibodies that are specific for TbMP63 and TbMP42 immunoprecipitate in vitro RNA editing activities. The proteins are present in the immunoprecipitates and sediment at 20S along with the in vitro editing, and RNA editing ligases TbMP52 and TbMP48. Recombinant TbMP63 and TbMP52 coimmunoprecipitate. These results indicate that these four proteins are components of the RNA editing complex and that TbMP63 and TbMP52 can interact.