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BackgroundEosinophilic granulomatosis with polyangiitis (EGPA) is characterized by eosinophilic inflammation of small with or without medium arteries. EGPA is a rare disease with varying prevalence and incidence rates globally. To date, limited information is available on the prevalence, incidence and burden of disease in the United Kingdom (UK).ObjectivesThe objectives were to estimate the prevalence and incidence of EGPA, and to describe the healthcare resource utilization (HCRU) among patients with EGPA in the UK.MethodsThis retrospective database study used the UK-based Clinical Practice Research Datalink (CPRD)-AURUM database linked to the Hospital Episode Statistics (HES). Prevalence was estimated from 2005 to 2019, and incidence was estimated from 2006 to 2019. HCRU was assessed in the 12-months following the first recorded diagnosis of EGPA (index date), and included hospitalizations, emergency room visits, procedures, outpatient specialist visits, primary care visits, and oral corticosteroid use.Results764 people were identified with EGPA in the UK. The prevalence of EGPA, reported in the database, increased from 22.7 to 45.6 per 1,000,000 persons from 2005 to 2019 (Figure 1), whereas the incidence of EGPA from 2006 to 2019 ranged from 2.28 to 4.00 per 1,000,000 person-years. 377 patients with EGPA were successfully linked to the CPRD-HES database. Patient characteristics were as follows: mean age (SD) was 57 years (14.2); 49% were male; 81% had asthma; and 11% had peripheral neuropathy prior to the index date. For patients with EGPA, 19% had an EGPA-related hospitalization and 50% had any-cause hospitalization within 1 year of the index date (Table 1). The mean length of stay was, 18 days and 16 days for EGPA-related and any-cause hospitalizations, respectively. 52% of patients with EGPA had undergone a medical procedure, 89% of patients with EGPA had an outpatient visit to a specialist. Almost all patients with EGPA visited a general practitioner within 1 year of their EGPA diagnosis (97%) and averaged 16.0 visits in 1 year. A significant proportion of the EGPA population were prescribed OCS; most EGPA patients had a prescription in the 0–3 months after the index date (64%), and patients on average had a prescription for OCS for 6 out of the 12 months after the index date.Table 1.HCRU among patients with EGPAHCRUNumber of patients N (%) [total days]Number of events per patient, Mean (SD)Total EGPA cohort (N)377 EGPA-specific hospitalizations72 (19.10)1.2 (1) EGPA-specific hospitalizations length of stay[1283]17.8 (23.3) Any-cause hospitalizations188 (49.87)1.7 (1) Any-cause hospitalizations length of stay[2992]15.9 (23.7) Any-cause A & E events19 (5.04)1.8 (2) Any-cause outpatient visits334 (88.59)9.8 (7) Any procedures undertaken196 (51.99)6.8 (6) General Practitioner visits366 (97.08)16.0 (11)A&E, Accident and Emergency; EGPA, eosinophilic granulomatosis with polyangiitis; HCRU, healthcare resource utilization.Figure 1.Prevalence of EGPA in the UK from 2005 to 2019Prevalence is expressed as cases per 1,000,000 persons. EGPA, eosinophilic granulomatosis with polyangiitis; UK, United Kingdom.ConclusionThe prevalence of EGPA increased over the study period in the UK, and the data show significant HCRU within 1 year of the first recorded diagnosis of EGPA. Almost all of the patients with EGPA were found to frequently visit the primary care physician and seek specialist care, and almost half required hospitalization. Funding: GSK [207888]AcknowledgementsFunding: GSK [207888]Disclosure of InterestsJeremiah Hwee Shareholder of: GSK, Employee of: GSK, Qinggong Fu Shareholder of: GSK, Employee of: GSK, Lorraine Harper Speakers bureau: Viopharm (2021), Roche (2017), Consultant of: GSK (2021), Viopharm (2021), Grant/research support from: Viopharm (researcher initiated project), MSD (researcher initiated project), Krishnarajah Nirantharakumar Consultant of: Boehringer Ingelheim (Consultancy on real world evidence), Grant/research support from: AstraZeneca, Vifor and Boehringer Ingelheim (Investigator led grants), Ruchika Goel: None declared, Rupert Jakes Shareholder of: GSK, Employee of: GSK

Lewy bodies and Lewy neurites are the defining neuropathological characteristics of Parkinson's disease and dementia with Lewy bodies. They are made of abnormal filamentous assemblies of unknown composition. We show here that Lewy bodies and Lewy neurites from Parkinson's disease and dementia with Lewy bodies are stained strongly by antibodies directed against amino-terminal and carboxyl-terminal sequences of alpha-synuclein, showing the presence of full-length or close to full-length alpha-synuclein. The number of alpha-synuclein-stained structures exceeded that immunoreactive for ubiquitin, which is currently the most sensitive marker of Lewy bodies and Lewy neurites. Staining for alpha-synuclein thus will replace staining for ubiquitin as the preferred method for detecting Lewy bodies and Lewy neurites. We have isolated Lewy body filaments by a method used for the extraction of paired helical filaments from Alzheimer's disease brain. By immunoelectron microscopy, extracted filaments were labeled strongly by anti-alpha-synuclein antibodies. The morphologies of the 5- to 10-nm filaments and their staining characteristics suggest that extended alpha-synuclein molecules run parallel to the filament axis and that the filaments are polar structures. These findings indicate that alpha-synuclein forms the major filamentous component of Lewy bodies and Lewy neurites.

Microtubule-associated protein tau is the major constituent of the paired helical filament, the main fibrous component of the neurofibrillary lesions of Alzheimer's disease. Tau is an axonal phosphoprotein in normal adult brain. In Alzheimer's disease brain tau is hyperphosphorylated and is found not only in axons, but also in cell bodies and dendrites of affected nerve cells. We report the production and analysis of transgenic mice that express the longest human brain tau isoform under the control of the human Thy-1 promoter. As in Alzheimer's disease, transgenic human tau protein was present in nerve cell bodies, axons and dendrites; moreover, it was phosphorylated at sites that are hyperphosphorylated in paired helical filaments. We conclude that transgenic human tau protein showed pre-tangle changes similar to those that precede the full neurofibrillary pathology in Alzheimer's disease.

The apolipoprotein E type 4 allele is a susceptibility gene for late-onset Alzheimer's disease. Apolipoprotein E is found in neurons, some of which contain paired helical filaments made of the microtubule-associated protein τ. Previous studies have demonstrated that the apoE3 isoform, but not the apoE4 isoform, binds τ with high avidity. Because the microtubule-associated protein MAP2c also effects microtubule assembly and stability, we examined interactions between apoE isoforms and MAP2c. Similar to the τ-binding results, apoE3, but not apoE4, bound MAP2c. Binding was detectable down to 10 −9 M MAP2c and 10 −8 M apoE3. Isoform-specific interactions of apoE with the microtubule-associated proteins MAP2c and τ might affect intracellular maintenance of microtubules and could contribute to a time-dependent pathogenesis of Alzheimer's disease.

Parkinson's disease (PD) is the most common neurodegenerative movement disorder (1). Neuropathologically, it is defined by nerve cell loss in the substantia nigra and the presence of Lewy bodies and Lewy neurites (2,3). In many cases, Lewy bodies are also found in the dorsal motor nucleus of the vagus, the nucleus basalis of Meynert, the locus coeruleus, the raphe nuclei, the midbrain Edinger-Westphal nucleus, the cerebral cortex, the olfactory bulb, and some autonomic ganglia (4).

Parkinson's disease (PD) is the most common neurodegenerative movement disorder (1). Neuropathologically, it is defined by nerve cell loss in the substantia nigra and the presence of Lewy bodies and Lewy neurites (2,3). In many cases, Lewy bodies are also found in the dorsal motor nucleus of the vagus, the nucleus basalis of Meynert, the locus coeruleus, the raphe nuclei, the midbrain Edinger-Westphal nucleus, the cerebral cortex, the olfactory bulb, and some autonomic ganglia (4).

Filamentous inclusions made of alpha-synuclein constitute the defining neuropathological characteristic of Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy. Rare familial cases of Parkinson's disease are associated with mutations A53T and A30P in alpha-synuclein. We report here the assembly properties and secondary structure characteristics of recombinant alpha-synuclein. Carboxy-terminally truncated human alpha-synuclein (1-87) and (1-120) showed the fastest rates of assembly, followed by human A53T alpha-synuclein, and rat and zebra finch alpha-synuclein. Wild-type human alpha-synuclein and the A30P mutant showed slower rates of assembly. Upon shaking, filaments formed within 48 h at 37 degrees C. The related proteins beta- and gamma-synuclein only assembled after several weeks of incubation. Synthetic human alpha-synuclein filaments were decorated by an antibody directed against the carboxy-terminal 10 amino acids of alpha-synuclein, as were filaments extracted from dementia with Lewy bodies and multiple system atrophy brains. Circular dichroism spectroscopy indicated that alpha-synuclein undergoes a conformational change from random coil to beta-sheet structure during assembly. X-ray diffraction and electron diffraction of the alpha-synuclein assemblies showed a cross-beta conformation characteristic of amyloid.

To facilitate studies of the normal biology of alpha-synuclein, a member of a family of neuronal proteins of unknown function, and to elucidate the role of alpha-synuclein pathologies in neurodegenerative diseases, we generated and characterized a panel of anti-synuclein antibodies. Here we demonstrate that these antibodies recognize defined epitopes spanning the entire length of human alpha-synuclein, and that some of these antibodies also cross-react with zebra finch and rodent synucleins. Since alpha-synuclein has been reported to be a major component of Lewy bodies (LBs) in Parkinson's disease (PD), dementia with LBs and common variants of Alzheimer's disease, we performed immunohistochemical studies showing that these antibodies label numerous LBs in the PD substantia nigra, thereby localizing protein domains throughout human alpha-synuclein in LBs. Taken together, our data indicate that this panel of antibodies can be exploited to probe the normal biology of alpha-synuclein as well as the role of pathological forms of this protein in PD and related neurodegenerative synucleinopathies.

Hyperphosphorylated microtubule-associated protein tau is the major proteinaceous component of the paired helical and straight filaments which constitute a defining neuropathological characteristic of Alzheimer's disease and a number of other neurodegenerative disorders. We have recently shown that full-length recombinant tau assembles into Alzheimer-like filaments upon incubation with heparin. Heparin also promotes phosphorylation of tau by a number of protein kinases, prevents tau from binding to taxol-stabilized microtubules, and produces rapid disassembly of microtubules assembled from tau and tubulin. Here, we have used the above parameters to study the interactions between tau protein and a number of naturally occurring and synthetic glycosaminoglycans. We show that the magnitude of the glycosaminoglycan effects is proportional to their degree of sulfation. Thus, the strongly sulfated glycosaminoglycans dextran sulfate, pentosan polysulfate, and heparin were the most potent, whereas the non-sulfated dextran and hyaluronic acid were without effect. The moderately sulfated glycosaminoglycans heparan sulfate, chondroitin sulfate, and dermatan sulfate had intermediate effects, whereas keratan sulfate had little or no effect. These in vitro interactions between tau protein and sulfated glycosaminoglycans reproduced the known characteristics of paired helical filament-tau from Alzheimer's disease brain. Sulfated glycosaminoglycans are present in nerve cells in Alzheimer's disease brain in the early stages of neurofibrillary degeneration, suggesting that their interactions with tau may constitute a central event in the development of the neuronal pathology of Alzheimer's disease.

PURPOSE This United Kingdom Medical Research Council (UK-MRC) study prospectively evaluated efficacy and long-term toxicity of adjuvant chemotherapy in high-risk stage I nonseminomatous germ cell tumors of the testis (NSGCTT). PATIENTS AND METHODS Eligible patients were those identified by the local histopathologist as having features confirmed in MRC surveillance studies to indicate an approximate 50% risk of relapse. Central histopathology review was undertaken. Chemotherapy consisted of two courses of cisplatin 100 mg/m2, bleomycin 30 mg weekly x 3, and etoposide 120 mg/m2 x 3, every 21 days (BEP). RESULTS One hundred fourteen eligible cases were enrolled. Median time of follow-up was 4 years, with 93 patients followed-up for at least 2 years. There have been two relapses, including one patient who did not have a germ cell tumor (GCT), according to the reference histopathologist. This patient is alive with active disease, the other has died. There was one death after a cerebrovascular accident during treatment. Assessment of fertility, lung function, and audiometry pretreatment and more than 9 months posttreatment indicated no clinically significant changes. A mean decrease in transfer factor coefficient (KCO) of 15% of the predicted value was noted, but no patient had symptomatic respiratory dysfunction. CONCLUSION There have been only two relapses among 114 cases of high-risk stage I NSGCTT treated with two courses of adjuvant BEP chemotherapy. The 95% confidence interval (CI) excludes a true relapse rate of more than 5%. Of 104 patients confirmed on histopathology review to have GCT, there has been only one relapse. Adjuvant chemotherapy is free from significant long-term toxicity, offering an effective alternative to surveillance or retroperitoneal lymph node dissection (RPLND) followed by surveillance, and may be preferred by some patients.

The apolipoprotein E (apoE) type 4 allele (APOE4) is a susceptibility gene for late-onset familial and sporadic Alzheimer disease. ApoE is found in some neurofibrillary tangle-bearing neurons, one of the major pathologic hallmarks of the disease. Neurofibrillary tangles contain paired helical filaments formed from hyperphosphorylated microtubule-associated protein tau. In vitro, tau binds avidly to apoE3, but not to apoE4, forming a bimolecular complex. Tau phosphorylated with a brain extract does not bind either isoform. ApoE3 binds to the microtubule-binding repeat region of tau, which is also the region that is thought to cause self-assembly into the paired helical filament. Binding studies with fragments of ApoE demonstrate that the tau-binding region of apoE3 corresponds to its receptor-binding domain and is distinct from the region that binds lipoprotein particles or beta/A4 peptide. Isoform-specific interactions of apoE with tau may regulate intraneuronal tau metabolism in Alzheimer disease and alter the rate of formation of paired helical filaments and neurofibrillary tangles.

A genetic classification of Alzheimer disease(s) (AD) is presented. We describe a potential metabolic process in individuals who inherit apolipoprotein E-epsilon 4 (APOE4, gene; apoE4, protein) alleles, leading to increased risk and earlier age of onset of late-onset Alzheimer disease. Apolipoprotein E-epsilon 3 (apoE3) binds to tau protein, possibly slowing the initial rate of tau phosphorylation and self-assembly into paired helical filaments (PHFs); apoE4 does not bind tau. Tau promotes microtubule assembly and stabilizes microtubules; hyperphosphorylated tau does not bind, thereby destabilizing microtubules. Hyperphosphorylated tau may self-assemble into PHFs. Over time a bias toward destabilization of microtubules and the formation of neurofibrillary tangles may occur in individuals who inherit APOE4 alleles, leading to a shorter functional neuronal life span. This hypothesis focuses attention on two important aspects of AD research design: (1) Although the inheritance of APOE4 is associated with increased risk and decreased age of onset, apoE4 does not directly cause the disease. Our data point to the absence of an important function of apoE3 or apoE2 in individuals who do not inherit these alleles as the genetically relevant metabolic factor. This has important implications for design of experiments directed toward understanding the relevant neuronal metabolism. (2) Should this hypothesis be proven and confirmed, targets for pharmaceutical therapy designed to mimic the metabolic function of apoE3 or apoE2 become a realistic preventive strategy.

Tau is a neuronal phosphoprotein whose expression is developmentally regulated. A single tau isoform is expressed in fetal human brain but six isoforms are expressed in adult brain, with the fetal isoform corresponding to the shortest of the adult isoforms. Phosphorylation of tau is also developmentally regulated, as fetal tau is phosphorylated at more sites than adult tau. In Alzheimer disease, the six adult tau isoforms become abnormally phosphorylated and form the paired helical filament, the major fibrous component of the characteristic neurofibrillary lesions. We show here that Ser-202 (in the numbering of the longest human brain tau isoform) is a phosphorylation site that distinguishes fetal from adult tau and we identify it as one of the abnormal phosphorylation sites in Alzheimer disease. The abnormal phosphorylation of tau at Ser-202 in Alzheimer disease thus recapitulates normal phosphorylation during development.

Es handelt sich bei den beschriebenen Fatienten um Einzelkasuistiken. Eine Einschatzung der Prognose der operierten Patienten oder der Wertigkeit des Verfahrens ist aufgrund der kurzen Beobachtungszeit und der geringen Patientenzahl noch nicht moglich.

From 7/1989 to 5/1993478 men with seminoma stage I (Tl–T3; no ipsilateral inguinal operation prior to orchiectomy) have been included in a phase III study assessing the field size of abdominal radiotherapy (30 Gy/3 wks). Group 1: paraaortic (p.a.) field from Thll to L5, the lateral borders including the processus transversi. Group 2: “dog-leg field” (p.a. + ipsilateral iliac field to the mid obturator level). Results With a median follow-up time of 29 months, 114 relapses have occurred, two of them within the pelvic area (2 years relapse-free survival 97%; 95% C.I./96%, 99%). Relapses were diagnosed between 4 months and 3 years from start of radiotherapy. Acute grade ≥3 gastrointestinal toxicity was observed in 11% and 17% of the patients with p.a. and dog-leg field, respectively. In Group I grade ≥2 myelotoxicity occurred in 5% of the patients and in 13% ofthe patients from Group 2.8 patients developed peptic ulcer after radiotherapy. After 1 year 11% patients in Group 1 and 30% of those in Group 2 were azoospermic. Conclusions In patients with seminoma stage I (T1–T3) and with undisturbed lymph node drainage of the primary tumour adjuvant radiotherapy to the p.a. lymph nodes is associated with reduced gastrointestinal, haematological and gonadal toxicity compared to standard pelvic + p.a. irradiation. The recurrence rate is low with either field.

The amino terminal sequence of the myosin light chain ( M r = 20 000) isolated from chicken gizzards was found to be acetyl-Ser-Ser-Lys-Arg-Ala-Lys-Ala-Lys-Thr-Thr-Lys-Lys-Arg-Pro-Gln-Arg-Ala-Thr-Ser(P)-Asn-Val-Phe. This sequence assignment differs from that reported by Maita et al. [(1981) European J. Biochem. 117, 417] in the order of the tryptic peptides. The revised amino acid sequence exhibits greater homology with the phosphorylation site sequences of the regulatory light chains from cardiac and skeletal muscle. Moreover it is now apparent why synthetic peptides corresponding to the previously reported sequence were very poor substrates for the myosin light chain kinase.

Extraction of bovine pituitaries at pH 7.0, in the presence or absence of protease inhibitors (PMSF, leupeptin, pepstatin A and EDTA) yielded both basic and acidic FGF components that were characterized by Western blotting and sequence analysis. Basic FGF comprised several components: an 18 KDa form that is similar, if not identical, to the basic FGF (1-146) already described; a 17 KDa form that is likely to be a new truncated molecular species (11-146) and a group of immunoreactive components of about 29 KDa. Acidic FGF showed several active components of pI 4.5-6.5. The most active component has a pI of approximately 5.0; molecular weight of 17 KDa and is shown, by Western blotting, to be similar to a truncated form of bovine brain acidic FGF. The biological activity of the latter component is shown to be neutralized by anti-brain acidic FGF antiserum.

Calcium is know to play a central role in controlling many cellular processes, for example, in secretion, in the visual cycle and in motility and has been implicated together with cyclic AMP in development, and in the control of many metabolic pathways; however, its role in regulating contraction in muscle is probably the best understood. Muscular contraction is controlled by specific calcium receptor proteins located on either the thick or thin filaments which are involved in switching actin-myosin interaction ON or OFF in response to changes in calcium concentration. In vertebrate striated and cardiac muscles, the regulatory proteins troponin and tropomyosin are associated with the thin filaments (Fig. 1). The extensive electron microscope, X-ray and biochemical evidence suggests that calcium induced changes in the troponin complex may be transmitted via a movement of the tropomyosin in such a way that it affects those sites in the actin molecules which are required for interaction with the myosin cross-bridges (see reviews Weber and Murray, 27; Huxley, 13). In molluscan muscles, the troponin complex is absent and instead calcium regulation of contractile activity requires the presence of specific regulatory light chains on the myosin (Fig. 1) (15, 24). However, little is known about how these regulatory light chains, under calcium, control, effect the transition of the myosin cross-bridge from the resting to the active state.

A 3300-base segment of Escherichia coli chromosomal DNA, cloned into pBR322, will complement a methionine auxotroph in which the lesion is a defective methionyl-tRNA synthetase with a much reduced affinity for methionine. Crude extracts of these transformants contain elevated levels of a protein which has a subunit molecular weight of 66 000, methionyl-tRNA synthetase aminoacylation activity in vitro and which cross-reacts with anti-(methionyl-tRNA synthetase) antibodies. This polypeptide is very slightly larger than the well-characterised and crystallised tryptic fragment of methionyl-tRNA synthetase. A DNA sequence of 1750 residues at one end of the cloned insert codes for a non-terminated open reading frame in which we can locate a large number of methionyl-tRNA synthetase tryptic and chymotryptic peptides. We have also sequenced 300 nucleotides upstream of this coding segment where we find a large invert repeat in the putative methionyl-tRNA synthetase promoter region.

EXCEPT when isotopes have been used, the investigation of the metabolism of the sugar phosphates and related compounds in animal and plant tissues has long been restricted by a lack of specific methods of determination1. Recently, however, specific enzymatic techniques have been developed for the estimation of both sugar phosphates and glycolytic intermediates in extracts from animal and plant tissues2–7. The enzymatic methods present greater difficulties with plant than with animal extracts, partly because the contents of phosphate compounds are smaller, and partly because the presence of pigments and inhibitors interferes with the assays. But successful results have now been obtained with extracts of potato tubers, strawberry leaves and fruits of banana, tomato and apple.