Chaperone therapy update: Fabry disease,GM1-gangliosidosis and Gaucher disease

Chaperone therapy is a newly developed molecular therapeutic approach to lysosomal diseases, a group of human genetic diseases causing severe brain damage. Based on early molecular studies during the last decade of the 20th century and early years of the 21st century, mainly on Fabry disease and G_M1-gangliosidosis, we found some mutant enzyme proteins were unstable in the cell, and unable to express catalytic activities. Subsequently galactose and other active-site binding substrate analogs were found stabilized and enhance the mutant enzyme activity in culture cells. We concluded that the mutant misfolding enzyme protein and substrate analog competitive inhibitor (chemical chaperone) form a stable complex to be transported to the lysosome, to restore the catalytic activity of mutant enzyme after spontaneous dissociation under the acidic condition. This gene mutation-specific molecular interaction is a paradoxical phenomenon that an enzyme inhibitor in vitro serves as an enzyme stabilizer in situ. First we developed a commercially available compound 1-deoxygalactonojirimycin (DGJ) for Fabry disease, and confirmed the above molecular phenomenon. Currently DGJ has become a new candidate of oral medicine for Fabry disease, generalized vasculopathy involving the kidneys, heart and central nervous system in the middle age. This drug development has reached the phase 3 of human clinical study. Then we found two valienamine derivatives, N-octyl-4-epi-β-valienamine (NOEV) and N-octyl-β-valienamine (NOV), as promising therapeutic agents for human β-galactosidase deficiency disorders (G_M1-gangliosidosis and Morquio B disease) and β-glucosidase deficiency disorders (phenotypic variations of Gaucher disease), respectively. Originally NOEV and NOV had been discovered as competitive inhibitors, and then their paradoxical bioactivities as chaperones were confirmed in cultured fibroblasts from patients with these disorders. Subsequently G_M1-gangliosidosis model mice have been used for confirmation of clinical effectiveness, adverse effects and pharmacokinetic studies. Orally administered NOEV entered the brain through the blood–brain barrier, enhanced β-galactosidase activity, reduced substrate storage, and improved neurological deterioration clinically. Computational analysis revealed pH-dependent enzyme-chaperone interactions. Our recent study indicated chaperone activity of a new DGJ derivative, MTD118, for β-galactosidase complementary to NOEV. NOV also showed the chaperone effect toward several β-glucosidase gene mutants in Gaucher disease. Furthermore a commercial expectorant drug ambroxol was found to be a chaperone for β-glucosidase. A few Gaucher patients responded to this drug with remarkable improvement of oculomotor dysfunction and myoclonus. We hope chaperone therapy will become available for some patients with Fabry disease, G_M1-gangliosidosis, Gaucher disease, and other lysosomal storage diseases particularly with central nervous system involvement.     

Chaperone therapy for lysosomal and non-lysosomal protein misfolding diseases

Chaperone therapy was introduced first as a new molecular therapeutic approach to lysosomal diseases. In a recent article, I reviewed the development of chaperone therapy mainly for lysosomal diseases. Then, more data have been collected particularly on non-lysosomal protein misfolding diseases. In this short review, I propose the concept of chaperone therapy to be classified into two different therapeutic approaches, for pH-dependent lysosomal, and pH-independent non-lysosomal protein misfolding diseases. The concept of lysosomal chaperone therapy is well established, but the non-lysosomal chaperone therapy is heterogeneous and to be investigated further for various individual diseases. As a whole, these two-types of new molecular therapeutic approaches will make an impact on the treatment of a wide range of pathological conditions caused by protein misfolding, not necessarily lysosomal but also many non-lysosomal diseases caused by gene mutations, metabolic diseases, malignancy, infectious diseases, and aging. The concept will open a completely new aspect of protein therapy in future.     

The French Pompe registry. Baseline characteristics of a cohort of 126 patients with adult Pompe disease

Pompe disease is a rare autosomal recessive muscle lysosomal glycogenosis, characterised by limb-girdle muscle weakness and frequent respiratory involvement. The French Pompe registry was created in 2004 with the initial aim of studying the natural history of French patients with adult Pompe disease. Since the marketing in 2006 of enzyme replacement therapy (alglucosidase alfa, Myozyme^®), the French Pompe registry has also been used to prospectively gather the biological and clinical follow-up data of all adult patients currently treated in France. This report describes the main clinical and molecular features, at the time of inclusion in the French registry, of 126 patients followed up in 21 hospital-based neuromuscular or metabolic centres. Sixty-five men and 61 women have been included in the registry. Median age at inclusion was 49 years, and the median age at onset of progressive limb weakness was 35 years. Fifty-five percent of the patients were walking without assistance, 24% were using a stick or a walking frame, and 21% were using a wheelchair. Forty-six percent of the patients needed ventilatory assistance, which was non-invasive in 35% of the cases. When performed, muscle biopsies showed specific features of Pompe disease in less than two-thirds of the cases, confirming the importance of acid alpha-glucosidase enzymatic assessment to establish the diagnosis. Molecular analysis detected the common c.-32-13T > G mutation, in at least one allele, in 90% of patients. The French Pompe registry is so far the largest country-based prospective study of patients with Pompe disease, and further analysis will be performed to study the impact of enzyme replacement therapy on the progression of the disease.     

Variantes del gen ABCB1 como factores de riesgo y factores moduladores de la edad de inicio en pacientes mexicanos con enfermedad desmielinizante

IntroductionThe C1236T, G2677T/A, and C3435T variants of the ABCB1 gene alter the functioning of P-glycoprotein and the transport of endogenous and exogenous substances across the blood-brain barrier, and act as risk factors for some neurodegenerative diseases.This study aimed to determine the association between demyelinating disease and the C1236T, G2677T/A, and C3435T variants of ABCB1 and its haplotypes and combinations of genotypes.MethodsPolymerase chain reaction with restriction fragment length polymorphism analysis (PCR-RFLP) and Sanger sequencing were used to genotype 199 patients with demyelinating disease and 200 controls, all Mexicans of mixed race; frequencies of alleles, genotypes, haplotypes, and genotype combinations were compared between patients and controls. We conducted a logistic regression analysis and calculated chi-square values and 95% confidence intervals (CI); odds ratios (OR) were calculated to evaluate the association with demyelinating disease.ResultsThe TTT and CGC haplotypes were most frequent in both patients and controls. The G2677 allele was associated with demyelinating disease (OR: 1.79; 95% CI: 1.12-2.86; P = .015), as were the genotypes GG2677 (OR: 2.72; 95% CI: 1.11-6.68; P = .025) and CC3435 (OR: 1.82; 95% CI: 1.15-2.90; P = .010), the combination GG2677/CC3435 (OR: 2.02; 95% CI, 1.17-3.48; P = .010), and the CAT haplotype (OR: 0.21; 95% CI: 0.05-0.66; P = .001).TTTTTT carriers presented the earliest age of onset (23.0 ± 7.7 years, vs. 31.6 ± 10.7; P = .0001).ConclusionsThe GG2677/CC3435 genotype combination is associated with demyelinating disease in this sample, particularly among men, who may present toxic accumulation of P-glycoprotein substrates.In our study, the G2677 allele of ABCB1 may differentially modulate age of onset of demyelinating disease in men and women.     

TRPV1 desensitisation and endogenous vanilloid involvement in the enhanced analgesia induced by capsaicin in inflamed tissues

The intraplantar acute administration of 10 μg of capsaicin to mice which had received complete Freund's adjuvant (CFA) 1 week before inhibits the thermal inflammatory hyperalgesia it induces and even produces a long-lasting analgesia for at least 2 weeks. In this study, we show that the administration of capsaicin (10 μg) also reduces the immediate licking behavior evoked by the intraplantar administration of a lower dose of capsaicin (0.1 μg), the duration of this inhibitory effect being greater in CFA-inflamed mice (at least 2 weeks) than in non-inflamed animals (less than 4 days). Since this reduction of capsaicin-induced licking behavior may be interpreted as a consequence of the transient receptor potential vanilloid 1 receptor (TRPV1) unresponsiveness, we conclude that the administration of 10 μg of capsaicin into inflamed tissues can render the TRPV1 desensitised. We next explored whether endogenous vanilloids released during inflammation contribute to maintain the analgesia triggered by exogenous capsaicin. The acute administration of capsazepine (10 μg; intraplantarly (i.pl.)) abolished the analgesic effect induced by the injection of capsaicin 1 week before in inflamed mice. From these results, it may be proposed that the maintenance by endovanilloids of the TRPV1 desensitisation induced by capsaicin could contribute to prolonging the analgesic effect induced by this agonist in inflamed tissues.     

Effect of systemic kynurenine on cortical spreading depression and its modulation by sex hormones in rat

BackgroundThe aura symptoms in migraine are most likely due to cortical spreading depression (CSD). CSD is favored by NMDA receptor activation and increased cortical excitability. The latter probably explains why migraine with aura may appear when estrogen levels are high, like during pregnancy. Kynurenic acid, a derivative of tryptophan metabolism, is an endogenous NMDA receptor antagonist whose cerebral concentrations can be augmented by systemic administration of its precursor l-kynurenine.ObjectiveTo determine if exogenous administration of l-kynurenine is able to influence KCl-induced CSD in rat, if the effect is sex-dependent and if it differs in females between the phases of the estrous cycle.MethodsAdult Sprague–Dawley rats (n = 8/group) received intraperitoneal (i.p.) injections of l-kynurenine (L-KYN, 300 mg/kg), L-KYN combined with probenecid (L-KYN + PROB) that increases cortical concentration of KYNA by blocking its excretion from the central nervous system, probenecid alone (PROB, 200 mg/kg) or NaCl. Cortical kynurenic acid concentrations were determined by HPLC (n = 7). Thirty minutes after the injections, CSDs were elicited by application of 1 M KCl over the occipital cortex and recorded by DC electrocorticogram. In NaCl and L-KYN groups, supplementary females were added and CSD frequency was analyzed respective to the phases of the estrous cycle determined by vaginal smears.ResultsIn both sexes, PROB, L-KYN and L-KYN + PROB increased cortical kynurenic acid level. PROB, L-KYN and L-KYN + PROB with increasing potency decreased CSD frequency in female rats, while in males such an effect was significant only for L-KYN + PROB. The inhibitory effect of L-KYN on CSD frequency in females was most potent in diestrus.Conclusionl-Kynurenine administration suppresses CSD, most likely by increasing kynurenic acid levels in the cortex. Females are more sensitive to this suppressive effect of l-kynurenine than males. These results emphasize the role of sex hormones in migraine and open interesting novel perspectives for its preventive treatment.     

A meta-analytic review of the correlation between peripheral oxytocin and cortisol concentrations

The stress dampening effects of exogenous oxytocin in humans have been well documented. However, the relation between endogenous oxytocin and cortisol is poorly understood. We conducted a meta-analysis on the correlation between oxytocin and cortisol levels measured at baseline (k = 24, N = 739). The effect size for the baseline correlation statistic was small (Pearson r = 0.163, p = 0.008), with high heterogeneity (I² = 67.88%). Moderation analysis revealed that studies where participants anticipated an experimental manipulation evidenced a greater positive correlation compared to those that did not (Pearson r = 0.318, p = 0.006). A supplementary analysis including additional studies indicated that oxytocin levels in unextracted samples were 60 times higher when using this questionable practice. The findings suggest that the interplay between oxytocin and cortisol is dynamic and sensitive to the anticipation of stress or novelty. Furthermore, extraction of oxytocin appears to be an essential methodological practice.     

The melatonin analog 5-MCA-NAT increases endogenous dopamine levels by binding NRH:quinone reductase enzyme in the developing chick retina

NRH:quinone reductase (QR2) is present in the retinas of embryonic and post-hatched (PH) chicks. 5-Methoxycarbonylamino-N-acetyltryptamine (5-MCA-NAT) is a QR2 ligand that increases cAMP levels in developing retinas, but it does not affect cAMP levels in CHO-QR2 cells. The dopamine quinone reductase activity of QR2 retrieves dopamine, which increases cAMP levels in developing retinas. The objective of the present study was to investigate whether 5-MCA-NAT increases endogenous dopamine levels in retinas from chick embryos and post-hatched chicks. Endogenous dopamine was measured by enzyme-linked immunosorbent assay (ELISA). 5-MCA-NAT increased retinal endogenous dopamine levels at all developmental stages studied and in PH chicks (−log EC50 = 11.62 ± 0.34 M). This effect was inhibited by non-selective antagonists of receptors and melatonin binding sites N-acetyl-2-benzyltryptamine (luzindole, 5 μM), but it was not inhibited by the Mel1b melatonin receptor antagonist 4-phenyl-2-propionamidotetralin (4-P-PDOT, 10 nM). The QR2 cosubstrate, N-methyl-dihydronicotinamide (NMH) (−log EC50 = 6.74 ± 0.26 M), increased endogenous dopamine levels in controls and in retinas stimulated with 5-MCA-NAT (3 nM). The QR2 inhibitor benzo[e]pyrene inhibited endogenous dopamine levels in both control (−log IC50 = 7.4 ± 0.28 M) and NMH-stimulated (at 100 nM and 1 μM benzo[e]pyrene concentrations) retinas. Theoretical studies using Molegro Virtual Docking software corroborated these experimental results. We conclude that 5-MCA-NAT increases the level of endogenous dopamine via QR2. We suggest that this enzyme triggers double reduction of the dopamine quinone, recovering dopamine in retinal development.     

Encéphalopathie mitochondriale neuro-gastro-intestinale (MNGIE) : quand et comment l’évoquer devant une anorexie mentale atypique ?

IntroductionThe Mitochondrial Neurogastrointestinal Encephalopathy (MNGIE) disease is an extremely underrated syndrome beginning around the age of eighteen years. Because of its severity, this diagnosis should be considered when a patient presents an atypical anorexia nervosa. MNGIE disease is inherited in an autosomal recessive manner and related to mutations of the TYMP gene (ch22q13.32-qter), encoding the thymidine phosphorylase. The MNGIE is often misdiagnosed and is associated with a time to diagnostic of about 12 years after first symptoms. Thus this critical review aims to help clinicians better identify symptoms and paraclinical markers of the MNGIE as a differential diagnosis of atypical anorexia nervosa.MethodsA literature search was performed using PubMed and Google Scholar databases.ResultsThe clinical diagnosis of the MNGIE disease should be based on the association of severe loss of weight and some additional symptoms: (1) severe gastrointestinal dysmotility (nausea, vomiting, intestinal pseudo-obstruction), (2) ptosis or external ophtalmoplegia and (3) peripheral sensorimotor neuropathy. When MNGIE disease is clinically suspected, paraclinical testing can help to validate the MNGIE diagnostic: (1) Arterial blood test reveals lactic acidemia (e.g. an increased serum concentration of lactate without pH modifications), and (2) Brain MRI indicates leukoencephalopathy, usually asymptomatic. Direct evidence of MNGIE disease is based on specific testing of: (1) the thymidine phopshorylase enzyme activity in leukocytes is less than 10% of the control, (2) the increase of plasmatic thymidine (> 3 μmol/L) and the increase of plamatic deoxyuridine (> 5 μmol/L), (3) the evidence of mutations of the TYMP gene by molecular genetic testing.ConclusionThe MNGIE disease is a severe trouble with multisystemic complications. The thymidine phopshorylase enzyme activity in leukocytes should be measured as soon as possible when a patient presents atypical anorexia nervosa.     

Developmental brain alterations in 17 year old boys are related to antenatal maternal anxiety

ObjectiveTo assess the association between maternal anxiety during pregnancy and the brain activity of 17 year old adolescents performing two cognitive control tasks.MethodsTwenty-three 17 year old boys of mothers whose level of anxiety was measured during pregnancy were investigated using ERP while performing a Go/Nogo paradigm assessing exogenous cognitive control and a Gambling paradigm requiring endogenous cognitive control.ResultsNo effects of antenatal maternal anxiety were observed in the Go/Nogo paradigm. However, in the Gambling paradigm adolescents of the high anxiety group (n = 8) showed a less efficient pattern of decision making compared to the adolescents in the low-average anxiety group (n = 15). Moreover, only for this task the ERP data showed an enlarged early frontal P2a component in the high anxiety group.ConclusionsThe brain activity of adolescents during an endogenous cognitive control task is associated to the level of anxiety experienced by their mother during pregnancy. This association was not observed during an exogenous cognitive control task.SignificanceThis study indicates that a child’s brain functionality is related to its mother’s anxiety during pregnancy. Endogenous cognitive control is regarded the cognitive function most affected by the level of antenatal maternal anxiety.     

Comprendre les troubles du sommeil de patients adultes atteints d’une maladie de Crohn pris en charge en ambulatoire

ObjectivesResearch has shown that sleep disturbances can negatively influence the progression of chronic inflammatory diseases, including chronic inflammatory bowel disease (IBD). More specifically, poor sleep quality is strongly related to the clinical activity of the disease. Nevertheless, some patients suffer from sleep disorders even when the disease is clinically inactive. Psychological factors, such as depression and anxiety, are also known to contribute to poor sleep quality. Depression and anxiety are common in chronic diseases. In addition, while the link between depression or anxiety and sleep disorders is well-known, the link between sleep disorders and inflammation has only been studied recently. Sleep studies in IBD patients have generally excluded patients with clinically diagnosed depression or anxiety in order to neutralize their effects on the relationship between inflammation and sleep disorders. Nevertheless, there is no consensus on the relationship between depression and anxiety and the clinical activity of the disease. When a patient with chronic inflammatory bowel disease (Crohn's disease here) complains of fatigue or poor sleep, the subjective aspects of these complaints therefore lead the clinician to consider them simply as: a characteristic of IBD, exhaustion related to the chronicity of the disease, unsatisfactory sleep quality, a manifestation of a depressive mood, or a consequence of an anxious state. When a patient reports a subjective complaint of poor sleep or fatigue in a complex, multi-determined clinical situation, it can thus be difficult to identify its most likely cause and to establish the best possible therapeutic intervention.Patients, materials and methodsThe aim of this work was to determine which element (disease activity, inflammation, depression, anxiety) is most closely related to sleep disorders in patients with Crohn's disease (CD) referred for outpatient psychological assessment. Ninety-seven patients with CD participated in this study. Their mean age was 34.70 (± 10.85) years. They were asked about their sleep (IQSP, ISI, ESD) and mood (HADS). They also provided details of clinical disease activity (Harvey–Bradshaw Index) and inflammation (CRP). In order to determine the nature and extent of the relationship between the variables, Spearman correlation coefficients were calculated, supplemented by multiple regression analyses to determine the variables that could explain the sleep disorders.ResultsThe results show that sleep quality (IQSP) was significantly predicted by the Harvey–Bradshaw score (β = 0.21; β standardized = 0.24; t = 2.6; P = 0.01) and depression (β = 0.45; β standardized = 0.41; t = 4.55; P < 0.001). The Harvey–Bradshaw score (β = 26; β standardized = 2; t = 2.27; P = 0.026) and depression score (β = 75; β standardized = 47; t = 5.34; P < 0.001) were related to the insomnia score (ISI). Finally, daytime sleepiness (DSA) was predicted by the depression score (β = 42; β standardized = 432; t = 3.17; P = 0.002) and by the CRP (β = − 0.05; β standardized = − 0.21; t = − 2.13; P = 0.036). The results show that the severity of clinical activity of the disease was associated with poor sleep quality and insomnia. However, there was a stronger association between the intensity of depression and sleep disturbances than between these variables and clinical disease activity. It therefore seems important that sleep disorders and their management should be considered first from the perspective of depression. However, it is important that CD is not assumed to be the sole cause of depression: other factors (dispositional or situational) should also be taken into consideration. Nevertheless, while our results show a weaker link between inflammation and sleep disorders than other studies, they confirm the link between sleep disorders and disease activity.ConclusionsIn order to predict the likelihood and nature of relapses, it seems important that future research should take into account not only disease activity and inflammation, but also disorders of arousal and nocturnal awakenings experienced by the patient.     

Calcitonin-gene related peptide and cerebral vasospasm

The pathophysiology of arterial vasospasm following subarachnoid hemorrhage (SAH) is poorly understood and the contribution of endogenous neuropeptides has not been sufficiently elucidated. Recently, we detected an excessive release of vasoconstrictive neuropeptide Y (NPY) in SAH patients and identified a significant correlation of NPY cerebrospinal fluid (CSF) levels with vasospasm-related ischemia. Here, we present the results of an experimental study on the possible role of the potent endogenous vasodilator calcitonin-gene related peptide (CGRP) in the acute stage of SAH. Twelve consecutive patients with SAH were included. Seven patients had severe arterial vasospasm, confirmed by transcranial doppler-sonography (TCD). Prospectively, CSF was collected from day 1 to day 10 after onset of the SAH. The levels of CGRP were determined in a competitive enzyme immunoassay and were correlated with the clinical course and hemodynamic changes. A cohort of 29 patients without CNS disease served as a control. CGRP was significantly higher in SAH patients compared with the control group (p < 0.05). From day 1 to day 4, the CGRP levels in patients without vasospasm were significantly higher than the levels of CGRP in patients with vasospasm (p < 0.05). These patients did not develop cerebral ischemia. The significantly increased levels of the CGRP during the first days after onset of the SAH in the non-vasospasm group indicate a potential protective role of CGRP. CGRP may alleviate arterial vasoconstriction and thus protect the brain from vasospasm and subsequent ischemia.     

Blood harmane (1-methyl-9H-pyrido[3,4-b]indole) concentration in dystonia cases vs. controls

BackgroundHarmane (1-methyl-9H-pyrido[3,4-b]indole) (HA) is a potent neurotoxin that has been linked to two neurological diseases, essential tremor and Parkinson's disease. Blood harmane concentrations [HA] are elevated in patients with both diseases. An important question is whether HA is specifically linked with these diseases or alternatively, is a non-specific marker of neurological illness.ObjectivesWe assessed whether blood [HA] was elevated in patients with a third neurological disease, dystonia, comparing them to controls.MethodsBlood [HA] was quantified by high performance liquid chromatography. Subjects comprised 104 dystonia cases and 107 controls.ResultsMean log blood [HA] in dystonia cases was similar to that of controls (0.41 ± 0.51 g⁻¹⁰/ml vs. 0.38 ± 0.61 g⁻¹⁰/ml, t = 0.42, p = 0.68). In unadjusted and adjusted logistic regression analyses, log blood [HA] was not associated with the outcome (diagnosis of dystonia vs. control): odds ratio (OR)_unadjusted = 1.11, 95% confidence interval (CI) = 0.69–1.79, p = 0.68; OR_adjusted = 1.07, 95% CI = 0.58–1.97, p = 0.84.ConclusionsIn contrast to the elevated blood [HA] that has been reported in patients with essential tremor and Parkinson's disease, our data demonstrate that blood [HA] was similar in patients with dystonia and controls. These findings provide the first support for the notion that an elevated blood [HA] is not a broad feature of neurological disease, and may be a specific feature of certain tremor disorders.     

Menstrual cycle and reproductive aging alters immune reactivity,NGF expression,antioxidant enzyme activities,and intracellular signaling pathways in the peripheral blood mononuclear cells of healthy women

Reproductive senescence in women is a process that begins with regular menstrual cycles and culminates in menopause followed by gradual development of diseases such as autoimmune diseases, osteoporosis, neurodegenerative diseases, and hormone-dependent cancers. The age-associated impairment in the functions of neuroendocrine system and immune system results in menopause which contributes to subsequent development of diseases and cancer. The aim of this study is to characterize the alterations in immune responses, compensatory factors such as nerve growth factor (NGF) and antioxidant enzyme activities, and the molecular mechanisms of actions in the peripheral blood mononuclear cells (PBMCs) of young (follicular and luteal phases), middle-aged, and old healthy women. Peripheral blood mononuclear cells were isolated from young women in follicular and luteal phases of the menstrual cycle (n = 20; 22.6 ± 2.9 yrs), middle-aged women (n = 19; 47.1 ± 3.8 yrs; perimenopausal) and old (n = 16; 63.2 ± 4.7 yrs; post-menopausal) women and analyzed for Concanavalin (Con A)-induced proliferation of lymphocytes and cytokine (IL-2 and IFN-γ) production, expression of NGF, p-NF-κB, p-ERK, p-CREB, and p-Akt, antioxidant enzymes [superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx), glutathione-S-transferase (GST)], extent of lipid peroxidation, and nitric oxide (NO) production. Serum gonadal hormones (17β-estradiol and progesterone) were also measured. A characteristic age- and menstrual cycle-related change was observed in the serum gonadal hormone secretion (estrogen and progesterone), T lymphocyte proliferation and IFN-γ production. Salient features include the age-related decline observed in target-derived growth factors (lymphocyte NGF expression), signaling molecules (p-ERK/ERK and p-CREB/CREB ratios) and compensatory factors such as the activities of plasma and PBMC antioxidant enzymes (SOD and catalase) and NO production. Further, an age-associated increase in p-NF-κB expression and lipid peroxidation was observed. Also, serum 17β-estradiol levels were positively correlated with IFN-γ production, SOD activity and NGF expression in the PBMCs. These results suggest that alterations in the levels of gonadal hormones are associated with immunosenescence characterized by decreased IFN-γ production and proliferation of T lymphocytes, decline in NGF expression, SOD and catalase activities, NO production, and signaling mechanisms and thus, may increase the incidence of diseases and cancer in women.     

Low-frequency repetitive transcranial magnetic stimulation for dyskinesia and motor performance in Parkinson’s disease

Dyskinesias are one of the most frequent and disabling complications of the long-term treatment of Parkinson’s disease (PD). Although the cause is not completely understood, it appears that an imbalance between excitatory and inhibitory inputs from the basal ganglia to the motor cortex leads to overactivation of motor and premotor areas. Overactivation of the supplementary motor area (SMA) has been observed in neuroimaging studies in dyskinetic PD patients. We investigated the effects of low-frequency repetitive transcranial magnetic stimulation (rTMS) of the SMA on levodopa-induced dyskinesias (LID) and motor performance in PD. We tested whether longer duration (10 days) and higher number of total pulses (1800 pulses) would enhance the beneficial effect. Seventeen dyskinetic PD patients were randomly assigned to real rTMS or sham (placebo) rTMS, and 1 Hz rTMS or sham rTMS was applied over the SMA for 10 consecutive days. Patients were assessed at baseline and 1 day after the last rTMS with a levodopa challenge test, and video recordings were taken. Dyskinesias and motor performance were rated off-line by two blinded raters using video recordings. After 10 days of treatment with rTMS, we observed that 1 Hz rTMS delivered over the SMA had decreased LID lasting for 24 hours without a change in motor performance, whereas sham rTMS induced no significant change in dyskinesia scores. These results support a possible therapeutic effect of low-frequency rTMS in LID. However, in order to suggest rTMS as an effective treatment, long-term observations and further investigations with a larger patient population are essential.     

Targeted high-resolution quadrupole-Orbitrap mass spectrometry analyses reveal a significant reduction of tachykinin and opioid neuropeptides level in PC1 and PC2 mutant mouse spinal cords

Tachykinin and opioid neuropeptides play a fundamental role in pain transmission, modulation and inhibition. The proteolysis control of endogenous tachykinin and opioid neuropeptides has a significant impact on pain perception. The role of proprotein convertases (PCs) in the proteolysis of proneuropeptides was previously established but very few studies have shown the direct impact of PCs on the regulation of specific tachykinin and opioid peptides in the central nervous system. There is an increasing interest in the therapeutic targeting of PCs for the treatment of pain but it is imperative to assess the impact of PCs on the pronociceptive and the endogenous opioid systems. The objective of this study was to determine the relative concentration of targeted neuropeptides in the spinal cord of WT, PC1^−/+ and PC2^−/+ animals to establish the impact of a restricted PCs activity on the regulation of specific neuropeptides. The analysis of tachykinin and opioid neuropeptides were performed on a HPLC-MS/MS (High-Resolution Quadrupole-Orbitrap Mass Spectrometer). The results revealed a significant decrease of Dyn A (p < 0.01), Leu-Enk (p < 0.001), Met-Enk (p < 0.001), Tach_58–71 (p < 0.05), SP (p < 0.01) and NKA (p < 0.001) concentrations in both, PC1^−/+ and PC2^−/+ animals. Therefore, the modulation of PCs activity has an important impact on specific pronociceptive peptides (SP and NKA), but the results also shown that endogenous opioid system is hindered and consequently it will affect significantly the pain modulatory pathways. These observations may have insightful impact on future analgesic drug developments and therapeutic strategies.     

Anticeramide antibody and butyrylcholinesterase in peripheral neuropathies

Ceramide is a glycosphingolipid, a component of nerve and non neuronal cell membrane and plays a role in maintaining the integrity of neuronal tissue. Butyrylcholinesterase (BChE) is a multifunctional enzyme, its involvement in neurodegenerative diseases has been well established. Anticeramide antibody (Ab-Cer) and enzyme BChE have been implicated in peripheral neuropathies. The present study investigates whether there is an association between Ab-Cer and BChE activities and peripheral neuropathies. Patients included: human immunodeficiency virus associated peripheral neuropathy (HIV-PN, n = 39), paucibacillary leprosy (PB-L, n = 36), multibacillary leprosy (MB-L, n = 52), diabetic neuropathy (DN, n = 22), demyelinating sensory motor polyneuropathy (DSMN, n = 13) and chronic inflammatory demyelinating polyneuropathy (CIDP, n = 10). Plasma Ab-Cer was measured by indirect enzyme linked immune assay (ELISA) and BChE activity in plasma was measured by colorimetric method. Ab-Cer levels were significantly elevated in MB-L and DN as compared to healthy subjects (HS). BChE levels were significantly higher in MB-L and DN as well as in HIV and HIV-PN. There is no significant difference in either Ab-Cer or BChE levels in DSMN and CIDP. Elevated plasma Ab-Cer and BChE levels may be considered significant in the pathogenesis of neuropathies. The variation in concurrent involvement of both the molecules in the neuropathies of the study, suggest their unique involvement in neurodegenerative pathways.     

Diseases of the circulatory system among adult people diagnosed with infantile autism as children: A longitudinal case control study

BackgroundResearch dealing with adult people with autism spectrum disorders (ASD) noticeably lags behind studies of children and young individuals with ASD.AimsThe objective of this study was to compare the prevalence and types of diseases of the circulatory system in a clinical sample of 118 adult people diagnosed with infantile autism (IA) as children with 336 sex and age matched controls from the general population.Methods and proceduresAll participants were screened through the nationwide Danish National Hospital Register. The average observation time of both groups was 37.2 years, and mean age at follow-up was 49.6 years.Outcomes and resultsOf the 118 people with IA, 11 (9.3%) were registered with at least one disease of the circulatory system against 54 (16.1%) in the comparison group (p = 0.09; OR = 0.54; 95% CI 0.3–1.2). Ischemic heart diseases occurred significantly more frequently among people in the comparison group (p = 0.02).Conclusions and implicationsIt is argued that diseases of the circulatory system may be underdiagnosed in people with IA because of the difficulties they face with respect to identifying and communicating symptoms of ill health. Bearing in mind that cardiovascular disease is the primary cause of death in most developed countries, it is suggested that to prevent disease and manage health conditions, health monitoring is essential in adult people with IA.     

Molecular basis of neurogenetic diseases

Molecular background of neurogenetic disease is briefly reviewed. Importance and usefulness of genetic testing are emphasized. Molecular genetics is a powerful tool for investigation of epileptic syndromes. Diagnosis based on gene analysis will give a new insight for pathophysiology and clinical outcome of the patient, and there is a hope to develop a new therapeutic approach in the near future. Among them a new molecular therapeutic trial for lysosomal diseases is being developed: chemical chaperone therapy. It will become a new approach to brain damage causing epilepsy and other phenotypic expressions of a large number of genetic diseases in the near future.     

Immune mediated conditions in autism spectrum disorders

We conducted a case-control study among members of Kaiser Permanente Northern California (KPNC) born between 1980 and 2003 to determine the prevalence of immune-mediated conditions in individuals with autism, investigate whether these conditions occur more often than expected, and explore the timing of onset relative to autism diagnosis. Cases were children and young adults with at least two autism diagnoses recorded in outpatient records (n = 5565). Controls were children without autism randomly sampled at a ratio of 5 to 1, matched to cases on birth year, sex, and length of KPNC membership (n = 27,825). The main outcomes – asthma, allergies, and autoimmune diseases – were identified from KPNC inpatient and outpatient databases. Chi-square tests were used to evaluate case-control differences. Allergies and autoimmune diseases were diagnosed significantly more often among children with autism than among controls (allergy: 20.6% vs. 17.7%, Crude odds ratio (OR) = 1.22, 95% confidence interval (CI) 1.13–1.31; autoimmune disease: 1% vs. 0.76%, OR = 1.36, 95% CI 1.01–1.83), and asthma was diagnosed significantly less often (13.7% vs. 15.9%; OR = 0.83, 95% CI 0.76–0.90). Psoriasis occurred more than twice as often in cases than in controls (0.34% vs. 0.15%; OR = 2.35, 95% CI 1.36–4.08). Our results support previous observations that children with autism have elevated prevalence of specific immune-related comorbidities.