Brain metal accumulation in Wilson's disease.

INTRODUCTION: Brain metal accumulation is suggested in the pathogenesis of numerous neurodegenerative disorders. In Wilson's disease (WD), only copper has been examined. The aim of the present study was to evaluate the copper, iron, manganese, and zinc concentrations in autopsy tissue samples from the brains of WD patients. METHODS: The study material consisted of 17 brains (12 WD patients, 5 controls) obtained at autopsy. Samples were taken from four different regions of each brain: frontal cortex, putamen, pons, and nucleus dentatus. The copper, manganese, and zinc content were determined using inductively coupled plasma mass spectrometry, and iron was assessed using flame atomic absorption spectroscopy. The results were analyzed according to select clinical variables. RESULTS: Copper content was increased homogenously in all investigated structures of the WD brains compared to controls (41.0 ± 18.6 µg/g vs.5.4 ± 1.8 µg/g; P<0.01). The mean concentrations of iron, manganese, and zinc were similar in WD and controls, but the iron level in the nucleus dentatus was higher in WD compared to controls (56.8 ± 14.1 µg/g vs. 32.6 ± 6.0 µg/g; P<0.05). Gender, age, and type and duration of WD treatment did not impact brain metals storage, but some correlations between the duration of the disease and copper and iron accumulation were observed. CONCLUSIONS: During the course of WD, copper accumulates equally in different parts of the brain. Zinc and manganese do not seem to be involved in WD pathology, but increased levels of iron were found in the nucleus dentatus. Thus, additional studies of brain iron accumulation in WD are needed.

Association of dopamine receptor gene polymorphisms with the clinical course of Wilson disease.

BACKGROUND: Dopamine receptor D2 (DRD2) polymorphisms are proposed to be important factors in the presentation of neuropsychiatric symptoms in many disorders, including decreased striatum levels of dopamine D2 receptors in Wilson disease. The present study investigated the association between DRD2 gene polymorphisms and clinical manifestation of Wilson disease. METHODS: Analyzing data from 97 symptomatic Wilson disease patients, we investigated the DRD2 gene polymorphisms rs1800497, rs1799732, and rs12364283. We assessed the polymorphisms impact on the phenotypic presentation of the disease. RESULTS: Generally, the DRD2 gene polymorphisms had no impact on the hepatic or neuropsychiatric clinical presentation of Wilson disease. However, rs1799732 deletion allele carriers with neuropsychiatric symptoms had earlier onset of WD symptoms by almost 6 years compared with individuals without this allele (22.5 vs. 28.3 years; P < 0.05). This unfavorable effect of the rs1799732 polymorphism was even more pronounced among adenosine triphosphatase 7B gene (ATP7B) p.H1069Q homozygous patients, in whom carriership of the deletion allele was related to earlier initial neuropsychiatric manifestation by 14 years (18.4 vs. 32.2 years; P < 0.01). CONCLUSIONS: Genetic variation of DRD2, specifically the rs1799732 polymorphism, may produce an earlier clinical presentation of Wilson disease neuropsychiatric symptoms and signs that occur in the course of dopaminergic system impairment due to copper accumulation in the brain. We speculate that this effect may be due to the impact of DRD2 polymorphism on dopamine D2 receptor density, but further studies are needed to understand the mechanisms of such phenotypic effects.

The effect of gender on brain MRI pathology in Wilson's disease.

Gender influence on the clinical manifestations of Wilson's Disease (WD) has been suggested; however, brain MRI pathology based on sexual dimorphism in WD has not yet been examined. The aim of this study was to analyse the effect of gender on brain MRI pathology according to the predominant form of WD. We retrospectively analysed the brain MR images of 204 newly diagnosed and untreated WD patients. The predominant form of the disease was neuropsychiatric (n = 105), hepatic (n = 67) or presymptomatic (n = 32). Overall, neuroimaging pathologies were found in 64.2 % WD patients. The clinical form analysis revealed significant gender-related differences. In the neuropsychiatric form, men presented with cerebellar atrophy and cortical brain atrophy more often than women (25/58 vs. 11/47; p < 0.05) and (23/58 vs. 12/47; p = 0.09), respectively. In contrast, women tended to present with globus pallidus lesions more often than men (25/47 vs. 20/58; p = 0.054). There were no gender differences observed in the hepatic form, but cortical brain atrophy presented more often in men than women (3/12 vs. 0/20; p < 0.05) in the presymptomatic form. According to our findings, there is a gender-dependent brain vulnerability to copper toxicity. We speculate that these differences are potentially related to an oestrogen protective effect and are due to differences in gender-related clinical forms.

Apolipoprotein E gene (APOE) genotype in Wilson's disease: impact on clinical presentation.

BACKGROUND: Wilson's disease (WD), an inherited copper metabolism disorder that leads to pathological tissue copper accumulation and secondary organ damage, is caused by mutations in the ATP-ase 7B gene (ATP7B). The apolipoprotein E gene (APOE) alleles ε2, ε3, and ε4 produce three different apoE isoforms with different biological effects, which can determine risks of many human diseases, including neurodegenerative and liver disease. This study aimed to evaluate the impact of APOE genotype on the variability of WD phenotypic expression. METHODS: We analyzed data on 383 WD consecutive patients in the WD registry. The APOE genotypes (APOE ε3/ε3 (wild-type), APOE ε2-positive, and APOE ε4-positive) were determined and the APOE genotype effect on the phenotypic WD presentation was assessed in all symptomatic WD patients, as well as in patient subgroups divided according to sex and ATP7B genotype. RESULTS: APOE genotype had no impact on WD presentation in the general population of symptomatic patients. However, APOE ε4-positive women tended to present WD symptoms earlier than women possessing the wild-type APOE ε3/ε3 genotype (24.2 vs. 27.9 years; p = 0.08). The effect of the APOE ε4-positive genotype was more pronounced in ATP7B p.H1069Q homozygous women, in whom disease symptoms started almost 6 years earlier (23.6 vs. 29.9 years; p < 0.05) than in APOE ε3/ε3 women. CONCLUSIONS: In women, APOE ε4-positive genotype is associated with earlier onset of WD symptoms, particularly among ATP7B p.H1069Q homozygous patients. Further studies are needed to understand the mechanisms of these gender-dependent phenotypic effects.

Gender differences in Wilson's disease.

BACKGROUND: Wilson's disease (WD) is an inherited disorder of copper metabolism. Although well documented in many disorders, gender hasn't been directly addressed in WD; therefore, our aim was to evaluate gender differences in WD. METHODS: We analyzed data on 627 consecutive WD patients entered into our registry (1958-2010). RESULTS: We observed a male predominance in our population of WD patients (327 males vs. 290 females; p<0.05). At disease diagnosis, 510/627 patients were symptomatic, most patients had the neuropsychiatric WD form (345/510; p<0.01). The neuropsychiatric form occurred predominantly in men versus women (209/278 vs. 136/232; p<0.01), especially the rigidity-tremor (71/111 vs. 40/111; p<0.05), rigidity (23/33 vs. 10/33; p=0.07) and psychiatric forms (46/71 vs. 25/71; p=0.06). The hepatic form occurred more frequently in women (96/165 vs. 69/165; p<0.01) and women developed the neuropsychiatric form 2 years later than men (29.4 vs. 27.1; p<0.05). CONCLUSIONS: According to our findings, the neuropsychiatric form of WD is predominant at diagnosis in both genders. The hepatic form of WD occurs more frequently in women, and women develop the neuropsychiatric form of disease almost 2 years later than men. We speculate these differences may be due to the protective effect of estrogens and iron metabolism differences.

The APOE polymorphism and 1-year outcome in ischemic stroke: genotype-gender interaction.

OBJECTIVE: In human genetic studies an effect of the apolipoprotein E gene (APOE) polymorphism on the risk, course and prognosis in chronic and acute nervous system disorders was established. We aimed to evaluate whether the APOE genotype is related to acute neurological impairments due to ischemic stroke (IS), and to outcomes (up to 1 year) indicated by severe functional disability, dependence in daily living or death. MATERIALS AND METHODS: A total of 657 patients (326 men, 331 women), divided into the three groups: E2 (APOEepsilon2/epsilon3 subjects), E3 (APOEepsilon3/epsilon3 subjects), and E4 (APOEepsilon3/epsilon4 and epsilon4/epsilon4 subjects), were analyzed. RESULTS: There was no association between the APOE genotype and baseline clinical characteristics, severity of neurological impairments during acute stroke, and 1-year outcome, when analyzing whole patient population. APOE gene interacted with gender in predicting severity of acute neurological deficit and post-stroke mortality within the period up to 1 year after the IS. Gender-stratified analysis indicated the E4 genotype as a significant independent positive predictor of death within 1 year after stroke incidence in men patients. CONCLUSION: Ischemic stroke severity and outcome may be affected by complex interactions between gender and genetic factors that warrant further exploration.

IL1RN intron 2 polymorphism caused by variable number tandem repeats is associated with 1-year outcome in patients with ischaemic stroke.

BACKGROUND: Results of experimental and clinical studies suggest that recombinant human interleukin 1 receptor antagonist (rhIL1ra) may be a good new therapeutic agent for acute stroke. In humans, IL1ra is encoded by the IL1RN gene located on chromosome 2. OBJECTIVES: To report on the association between different genotypes of the variable number tandem repeat (VNTR) polymorphism within IL1RN, and disease severity and 1-year outcome in patients with ischaemic stroke. METHODS: IL1RN genotypes were evaluated using polymerase chain reaction in 391 patients with ischaemic stroke diagnosed according to the World Health Organization definition. The effects of IL1RN genotypes on severity of stroke at maximum impairment, and on the survival status and neurological and functional condition of patients at 7 days, 1 month, 3 months and 1 year after the onset, were evaluated. RESULTS: No relationship was found between IL1RN genotypes and severity of symptoms at the time of maximum impairment. Homozygotes for the IL1RN*2 allele showed less severe neurological and functional impairments when assessed after the time period between 7 days and 1 year after stroke compared with carriers of the other two IL1RN genotypes. Patients with at least one copy of the IL1RN*2 allele had increased risk of death during the first week, and patients homozygotic for this allele had increased risk of death within the first month after stroke. CONCLUSION: IL1RN intron 2 variable number tandem repeats polymorphism influences the clinical outcome in patients with ischaemic stroke. It may possibly modify effects of treatment with rhIL1ra in patients with acute stroke.

Frameshift and nonsense mutations in the gene for ATPase7B are associated with severe impairment of copper metabolism and with an early clinical manifestation of Wilson's disease.

Wilson's disease (WD) is an autosomal recessive disorder of copper metabolism. The clinical phenotype of the disease is varied. It is proposed that this variation may be a result of differential functional disruption of ATPase7B (ATP7B) resulting from mutations in the gene ATP7B. We aimed to assess the relationship between specific mutational defects in ATP7B and divergence in the phenotypic expression of WD. One hundred and forty-two patients with clinically, biochemically and genetically diagnosed WD were included in the study. The phenotypic expression of WD was compared between patients with different types of mutations in ATP7B, detected by direct sequencing of exons 1-21 of the gene. Twenty-six mutations were identified in ATP7B; eleven of them were mutations predicted to result in the absence of a full-length normal protein [frameshift/nonsense mutations; classified as 'severe' mutations (SMs)], 14 were missense mutations (MMs) and one was a splice site mutation. Patients with one or two SMs on their alleles had lower serum copper and ceruloplasmin and were younger when the first symptoms of the disease appeared, compared with individuals with two MMs. The effect of SMs on the WD phenotype was dose-dependent. It is concluded that mutations within ATP7B are very heterogeneous. Frameshift and nonsense mutations are associated with a severe phenotype of WD.

Estrogen and cytokines production - the possible cause of gender differences in neurological diseases.

Naturally occurring sexual dimorphism has been implicated in the risk, progression and recovery from numerous neurological disorders. These include head injury, multiple sclerosis (MS), stroke, and neurodegenerative diseases (Parkinson's disease (PD), Alzheimer's disease (AD) or amyotrophic lateral sclerosis (ALS). Accumulating evidence suggests that observed differences between men and women could result from estrogen's wide range of effects within the mammalian central nervous system (CNS), with it's neuroprotective effect being one of the most important. It seems possible that neuroprotective activity of estrogen could be partially a result of it's anti-inflammatory action. It has been well established that inflammation plays an important role in the etiopathogenesis and manifestation of brain pathological changes. In this regard, an important role has been suggested for pro-inflammatory cytokines produced by activated glial cells, neurons and immune cells that invade brain tissue. Within the CNS, cytokines stimulate inflammatory processes that may impair blood-brain barrier permeability as well as promote apoptosis of neurons, oligodendrocytes and induce myelin damage. Given that estrogen may modulate cytokine expression, coupled with the fact that gender differences of cytokine production are apparent in animal models of PD and MS, suggests an important connection between hormonal-cytokine link in neurodegeneration. Indeed, while MS patients and mice subjected to experimental autoimmune encephalomyelitis (EAE) display gender specific alterations of IFN-gamma and IL-12, variations of TNF and IL-6 were associated with PD. Also in case of more acute neurodegenerative conditions, such as stroke, the effect of IL-6 gene G-174C polymorphism was different in males and females. Given that our understanding of the role of estrogen on cytokine production and accompanying CNS pathological conditions is limited, the present reviews aims to present some of our recent findings in this area and further evaluate the evidence that may be relevant to the design of new hormonal anti-inflammatory treatment strategies for neurodegenerative diseases.

Elevated levels of anti-heat shock protein antibodies in patients with cerebral ischemia.

One of the important mechanisms involved in the development of vascular lesions leading to ischemic stroke could be an immune response to heat shock proteins (hsp). For carotid atherosclerosis and myocardial infarction, an association with an increase in anti-hsp 65 antibodies has been demonstrated. The aim of our study was (1) to investigate whether ischemic stroke is associated with a humoral immune response to hsp; (2) to study the connection between anti-hsp antibodies and other stroke risk factors; (3) to estimate if the elevated levels of anti-hsp antibodies could be an independent risk factor for stroke. We examined 180 patients (in the first 48 h after stroke onset) and 64 age-matched healthy controls. The levels of IgG and IgM antibodies to hsp 65 and 70 were measured by ELISA. Ischemic stroke was connected with a significant elevation of anti-hsp 65 and anti-hsp 70 antibody levels (IgG and IgM) compared with controls (p < 0.0001). The multifactorial logistic regression analysis showed that increased levels of anti-hsp 65 and anti-hsp 70 IgG antibodies are independent risk factors for stroke. Our results suggest that humoral immunity to hsp is common in stroke patients and that elevated levels of anti-hsp antibodies could be triggering factors for stroke.

Frequency of His1069Gln and Gly1267Lys mutations in Polish Wilson's disease population.

Wilson's disease is an autosomal recessive disorder. More than 60 mutations of the Wilson's disease gene have been described so far. We have analysed 148 Polish Wilson's disease patients from 95 families for His1069Gln and Gly1267Lys mutations and correlated this finding with age and clinical form of the disease at presentation. To identify these mutations, single strand conformation polymorphism analysis was performed. In our group there were 94 patients with neurological presentation, 28 with hepatic presentation, whilst 26 were in a pre-clinical stage of the disease. His1069Gln mutation was present on 171 (57%) of the 296 studied chromosomes, and Gly1267Lys mutation was present on 27 chromosomes (9.1%). Most of our patients were homozygous or heterozygous for His1069Gln mutation (39.9% and 30.4%, respectively); 4% of the patients were homozygous for Gly1267Lys mutation and 5.4% had both of these described mutations on their chromosomes. His1069Gln and Gly1267Lys mutations occurred often in our Wilson's disease patient population but we did not find any relationship between investigated mutations and the clinical form of Wilson's disease or age of first symptoms.

[The relationship between immunological parameters with etiopathogenesis and clinical course of stroke]. / Zwiazek czynników immunologicznych z etiopatogeneza i przebiegiem klinicznym udaru mózgu.

We observed a lot of immune system disorders significantly influencing the development and clinical course of stroke. Depression of cell-mediated immune reactivity was observed in the early stage of stroke. It was manifested by the decrease of the number of T lymphocytes, depression in lymphocyte blastogenesis, diminished production of the migration inhibition factor and reduced delayed-type skin reactivity. Observed depression was probably caused by severe metabolic and endocrinological disorders often seen in the acute phase of the disease and related with increased patients' susceptibility to infections. Simultaneously we observed that the elevated total WBC was an independent stroke risk factor and predictor of 30-days stroke fatality. The activation of the adhesion molecules expression on granulocytes (CD18) and the increased cytokine production by leukocytes could favour leukocytes influence to the ischemic area and potentiate brain injury. The chronic inflammation could be also responsible for the development of vascular injuries leading to the ischemia. In our studies we demonstrated the high levels of antibodies to cardiolipins and heat shock proteins and the increased blood levels of immune complexes (i.c.). We observed the presence of Chlamydia pneumoniae and CMV antigens in isolated i.c. It suggests that different markers of chronic inflammation observed at very early stage of the disease are probably related with the chronic infection potentially leading to the development of atherosclerotic lesions and destabilization of atherosclerotic plaque.

[Presence of immune complexes in ischemic stroke]. / Obecnosc kompleksów immunologicznych u chorych z udarem niedokrwiennym mózgu.

There is increasing evidence that inflammatory responses play an important role in the pathogenesis of atherosclerosis and cerebral infarct. The aim of this study was to determine the amount of immunocomplexes (i.c.) in patients with stroke in the early period of the disease. Studies were performed on 35 subjects. The concentration of immunocomplexes was determined with the precipitation method (3.5% polyethylenoglikol was used). Increased concentration of i.c. was found in patients with cerebral infarct (after 12 hours and 7 days). It suggests, that i.c. could be one of the markers for systemic inflammation and be important in the patogenesis of atherosclerosis and stroke.

[Practical usefulness of a donor registry with determined HLA class I antigens--current personal experience]. / Praktyczne znaczenie rejestru dawców z okreslonymi antygenami HLA klasy I--dotychczasowe doswiadczenie wlasne.

The experience in organizing the HLA class I typed donors registry and practical application for platelet transfusions in immunized patients is presented. It appears that the registry containing 3461 donors gives quite a high possibility of finding a compatible or partly compatible HLA donor for a patient with relatively high frequency antigens. A discrepancy is, however, observed between finding a donor in the registry and the transfusion of the matched platelets. Better collaboration between physicians and persons organizing the registry is required to improve transfusions of the matched platelets in immunized patients. Preliminary experience is also described concerning the usefulness of the same registry for finding the HLA class I compatible donors for bone marrow transplantation.

[Evaluation of the microplate enzyme-linked assay with isolation of glycoprotein IIb/IIIa from platelet membranes: comparison with immunofluorescent tests and monoclonal antibodies]. / Ocena wykrywalnosci przeciwcial przeciwplytkowych przy uzyciu testu enzymatycznego z wyizolowana glikoproteina IIb/IIIa blony krwinki plytkowej: porównanie z testem immunofluorescencyjnym oraz testem enzymatycznym z uzyciem przeciwcial monoklonalnych.

The enzyme test with GPIIb/IIIa has been used to detect antibodies in 500 sera from patients with thrombocytopenia and mothers of infants with thrombocytopenia. The results were compared with antibody detection in the platelet suspension immunofluorescence test (PIFT) and in the monoclonal antibody immobilization of platelet antigens (MAIPA). Platelet antibodies were detected in 125 sera (25%). In 46.2% the results were positive in all three tests. However, in 38.4% the ELISA was negative while the PIFT and MAIPA were positive. On the other hand, in 14.4% the ELISA was positive while the PIFT and/or the MAIPA were negative. The ELISA appears to be useful in the detection of anti-HPA-1a antibodies, which are the main cause of AIMN and PTP. This test, however, seems to be less useful in the detection of autoantibodies. The significance of the ELISA in the detection of platelet alloantibodies responsible for refractoriness to platelet transfusions is not clear yet and requires further investigations.