Perampanel, an AMPA receptor antagonist: From clinical research to practice in clinical settings.

Epileptic seizures are refractory to treatment in approximately one-third of patients despite the recent introduction of many newer antiepileptic drugs (AEDs). Development of novel AEDs therefore remains a high priority. Perampanel is a first-in-class non-competitive selective AMPA receptor antagonist with a unique mechanism of action. Clinical efficacy and safety of perampanel as adjunctive treatment for focal seizures with/without secondary generalization (±SG) and primary generalized tonic-clonic (PGTC) seizures have been established in five phase 3 randomized controlled trials (RCTs), and a long-term extension study, and perampanel is approved as monotherapy for focal seizures ±SG in the USA. In patients with focal seizures ±SG, add-on perampanel resulted in median percent reduction in seizure frequency 23.3%-34.5% and ≥50% responder rate 28.5%-37.6%; in PGTC seizures, these results were 76.5% and 64.2%, respectively. Efficacy among adolescents (reduction in seizure frequency 34.8%-35.6%; ≥50% responder rate 40.9%-45.0%) and elderly people (reduction in seizure frequency 12.5%-16.9%; ≥50% responder rate 22.2%-42.9%) is similar to those in adults, and results remain comparable between Asian (reduction in seizure frequency 17.3%-38.0%) and global populations. Perampanel has been extensively studied in real-world clinical practice, with similar efficacy and safety results to the RCTs (≥50% responder rate 12.8%-75.0%; adverse events of somnolence/sedation, dizziness, ataxia, and behavioral changes). Real-world observational studies suggest that perampanel tolerability can be improved by slow titration (2 mg every 2-4 weeks), and bedtime administration can mitigate somnolence and dizziness. Counseling about the potential for behavioral changes and close monitoring are recommended.

Nonadverse Effects on Allergenicity of Isopentenyltransferase-Transformed Broccoli

Introducción: Los organismos modificados genéticamente (GM) dan lugar a mejoras en la agricultura moderna relacionados con la eficiencia y beneficios de la producción de alimentos, sin embargo el potencial impacto de estos en la salud humana no ha sido clarificado. El motivo de este estudio fue investigar la alergenicidad entre el brócoli modificado genéticamente mediante isopenteniltransferasa frente al no modificado. Para ello se utilizó suero de pacientes alérgicos a brócoli para identificar la alergenicidad de ambos brócolis modificado o no modificado. Se estudió la unión de la IgE al brócoli de uno u otro tipo mediante inmunoblotting, ELISA y liberación de histamina. Resultados: Los resultados mostraron que el 7.2% de los pacientes tenían reacciones a brócoli (B. oleracea). La IgE específica a brócoli y la IgE total de los sujetos sensibilizados mostraban una correlación. Mediante Western blot observamos una heterogenia en la IgE en el suero de los pacientes reactiva frente a los componentes alergénicos. La alergenicidad de los brócolis, modificado o no modificado, no mostró diferencias significativas, lo cual indica que la trasformación del brócoli no afecta a su alergenicidad. Conclusiones: En nuestro estudio demostramos la ausencia de diferencias entre brócoli trasformado genéticamente o no (isopenteniltransferasa), por lo tanto no son esperables efectos sobre la alergenicidad en el brócoli transformado (AU)

Background: Genetically modified organisms (GMOs) provide modern agriculture with improvements in efficiency and the benefits of enhanced food production; however, the potential impact of GMOs on human health has not yet been clarified. Objective: To investigate the allergenicity of isopentenyltransferase (ipt)-transformed broccoli compared with non-GM broccoli. Methods: Sera from allergic individuals were used to identify the allergenicity of GM and non-GM broccoli. Immunoglobulin (Ig) binding of different lines of GM and non-GM broccoli was identifi ed using immunoblotting, enzyme-linked immunosorbent assay, and the histamine release assay. Results: Positive reactions to broccoli (Brassica Oleracea) were observed in 7.02% of individuals. Specific IgE to broccoli and total IgE from allergic individuals were well correlated. The different tests performed showed no significant differences in the allergenicity of conventionally raised and GM broccoli, indicating the absence of unexpected effects on allergenicity in ipt-transformed plants. Using Western blot analysis, we detected heterogeneous IgE-reactive allergenic components in broccoli-allergic sera, but no significant differences between GM and non-GM broccoli were observed in serum from the same patients. Conclusions: Our study demonstrates that there are no differences between GM (ipt-transformed) broccoli and non-GM broccoli, as determined by specific IgE in sera from broccoli-allergic patients. This indicates that there were no unexpected effects on allergenicity in this GM broccoli (AU)

Immune Responses to Tyrophagus putrescentiae–Induced Airway Inflammation in Mice

Antecedentes: Los ácaros de almacenamiento son una fuente de aeroalérgenos en pacientes con rinitis y asma alérgica. El Tyrophagus putrescentiae (Tp) es causante de reacciones de hipersensibilidad de las vías respiratorias, sin embargo los mecanismos y la patogenia de esta enfermedad están aún por dilucidar. Objetivo: El presente estudio tuvo como objetivo el establecer un modelo murino de asma alérgica inducida por el Tp. Métodos: Los ratones fueron sensibilizados con un extracto crudo de Tp por vía intra-peritoneal y, posteriormente, provocados con el mismo, por vía intratraqueal. Se cuantificaron diferentes parámetros de la respuesta inmunitaria como: variaciones en la concentración de inmunoglobulinas, subpoblaciones leucocitarias, citocinas y expresión de genes, así como la función pulmonar y, finalmente, se realizaron estudios histológicos. Resultados: Los ratones sensibilizados y provocados con Tp desarrollaron un aumento significativo de los niveles de IgE e IgG1 específicas de Tp en suero, en comparación con el grupo NS (p <0,01). Asimismo, se observó un incremento significativo, después de la sensibilización, en la cifra de leucocitos inflamatorios (neutrófilos y eosinófilos) y de algunas citocinas (IL-4, IL-5 e IL-13). En la función pulmonar, se obtuvieron valores significativamente mayores de Penh (p <0,05) en los ratones sensibilizados a Tp. En el estudio de la expresión génica, se observó que los genes relacionados con la respuesta Th2 (IL-4, IL-5, IL-13, y RANTES), Th2 específica del factor de transcripción GATA-3, pro-inflamatoria (IL-6), y Th17 (IL-17F), aumentó significativamente tras la provocación intratraqueal. En los ratones sensibilizados con extracto crudo Tp se confirmó una histología pulmonar con inflamación del tejido pulmonar y alteraciones traqueales. Conclusión: La sensibilización intraperitoneal con extracto crudo de T. putrescentiae, seguida de una provocación intratraqueal, puede inducir la inflamación de las vías aéreas en ratones. Este modelo murino pudiera servir de base para evaluar la eficacia terapéutica de fármacos en la inflamación de las vías respiratorias, inducida por T. putrescentiae, dadas las similitudes encontradas en aspectos inmunológicos y clínicos, con el asma alérgica de los seres humanos (AU)

Background: Storage mites are a source of aeroallergens that affect patients with allergic rhinitis and asthma. Tyrophagus putrescentiae is a causative factor of airway hypersensitivity, but the mechanisms and pathogenesis of T putrescentiae–induced allergy are not well understood. Objective: This study aimed to develop a murine model of T putrescentiae–induced allergic asthma. Methods: Immune responses and physiologic variations in immunoglobulins (Ig), leukocyte subpopulations, cytokines, gene expression, pulmonary function, and lung pathology were evaluated after intraperitoneal sensitization and intratracheal challenge with crude extract of T putrescentiae. Results: After sensitization with aluminum hydroxide and challenge with T putrescentiae in mice, levels of T putrescentiae–specific IgE and IgG1 in sera increased significantly compared to the normal saline group (P<.01). Values for inflammatory leukocytes (neutrophils and eosinophils) and cytokines (interleukin [IL] 4, IL-5, and IL-13) increased significantly after sensitization. In terms of pulmonary function, pause values were significantly enhanced in T putrescentiae–sensitized mice after intratracheal challenge with T putrescentiae (P<.05). Expression of type 2 helper T cell (TH2)–related genes (IL4, IL5, IL13, and RANTES), TH2-specifi c transcription factor (GATA-3), and proinflammatory genes (IL6), and TH17-related genes (IL17F) increased significantly after airway challenge. Sensitization with T putrescentiae crude extract led to inflammation of lung tissue, thickening of the tracheal wall, and tracheal rupture. Conclusions: Intraperitoneal sensitization followed by intratracheal challenge with crude extract of T putrescentiae can induce airway inflammation in BALB/c mice. The symptoms observed in a mouse model of allergic asthma, in terms of immune and clinical parameters, are reminiscent of the symptoms of allergic asthma in humans. A mouse model can be used to evaluate the therapeutic effectiveness of drugs on T putrescentiae–induced airway inflammation in humans (AU)

Increased expression of PRL-1 protein correlates with shortened patient survival in human hepatocellular carcinoma

BACKGROUND: The purposes of the current study were to investigate whether overexpression of the PRL-1 is clinically relevant to hepatocellular carcinoma (HCC) and whether expression patterns of PRL-1 in HCC have diagnostic and prognostic value. METHODS: Immunohistochemistry analysis was performed for PRL-1 in 60 HCC samples. The data were correlated with clinicopathological features. The univariate and multivariate survival analyses were also performed to determine their prognostic significance. RESULTS: PRL-1 protein was overexpressed (83%) in HCC as compared with the adjacent normal tissue. PRL-1 expression was not influenced by chronic alcohol exposure or cirrhosis. High expression of PRL-1 was correlated with smoking (p=0.012), cirrhosis (p=0.047) and histological grade (p=0.055). The Kaplan-Meier survival curves showed that high PRL-1 expression related to a poor survival with statistical significance (I vs. III, p=0.010; II vs. III, p=0.001). Univariate analysis showed that PRL-1 expression was associated with tumour size, stage and PRL-1 score. Multivariate analysis revealed that the PRL-1 protein expression level was an independent factor for overall survival (HR, 5.367; 95% CI, 2.270-12.692; p=0.001). This is the first demonstration that the expression level of PRL-1 is correlated with tumour progression and prognosis in HCC. CONCLUSIONS: Along with other results, the PRL-1 protein is a candidate biomarker and a potential target for novel therapies against human HCC progression (AU)

Imiquimod simultaneously induces autophagy and apoptosis in human basal cell carcinoma cells.

BACKGROUND: Imiquimod shows antitumour activity through the stimulation of cell-mediated immunity in vivo. Recent studies have shown that imiquimod promotes apoptosis in melanoma cells and induces autophagy in macrophage cell lines. OBJECTIVES: To evaluate the imiquimod-induced apoptosis, autophagy and their relationship in a basal cell carcinoma (BCC) cell line. METHODS: Cell viability was determined by XTT test. Apoptosis was evaluated by DNA content assay, annexin V/propidium iodide staining assay and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling assay. Autophagy was determined by LC3 immunoblotting, EGFP-LC3 puncta formation and quantification of acidic vesicular organelles with acridine orange staining. The temporal and spatial differences of imiquimod-induced apoptosis and autophagy were examined by immunoblotting and simultaneously monitored by staining the EGFP-LC3 transfected cells with caspase 3 fluorogenic substrate. We inhibited the apoptosis and autophagy by pancaspase inhibitor and siRNA for Beclin 1 or Atg5, respectively, to evaluate the interplay between imiquimod-induced apoptosis and autophagy. RESULTS: We found that imiquimod induces autophagy and apoptosis in BCC cells in a time- and dose-dependent manner. Imiquimod not only induced EGFP-LC3 puncta formation for autophagy, but also simultaneously activated an apoptotic caspase cascade in the same cells. Both apoptosis and autophagy induced by imiquimod cooperate to cause BCC cell death. However, inhibition of imiquimod-induced apoptosis increased the strength of autophagy, and inhibition of imiquimod-induced autophagy further promoted cell apoptosis. CONCLUSIONS: This study not only demonstrates that imiquimod can directly induce autophagy and apoptosis in BCC cells, but also shows the cooperation and coordination between these two processes to induce cell death.

Factors associated with isolated anti-hepatitis B core antibody in HIV-positive patients: impact of compromised immunity.

In regions that are hyperendemic for chronic hepatitis B virus (HBV) infection, prevalence of and risk factors associated with isolated anti-hepatitis B core antibody (anti-HBc) in HIV-positive patients are less well described. HIV-positive patients who were tested for hepatitis B surface antigen (HBsAg), anti-hepatitis B surface antibody (anti-HBs) and anti-HBc at designated hospitals for HIV care in Taiwan were included for analysis. HBV DNA was detected by real-time polymerase chain reaction in patients with and without isolated anti-HBc. Of 2351 HIV-positive patients, 450 (19.1%) were HBsAg positive, 411 (17.5%) were anti-HBc positive alone and 963 (41.0%) for both anti-HBs and anti-HBc. Compared with patients who were positive for both anti-HBs and anti-HBc, patients with isolated anti-HBc were older, less likely to have anti-hepatitis C virus antibody (anti-HCV), had lower CD4 lymphocyte counts and higher plasma HIV RNA loads. Older age (adjusted odds ratio, 1.029; 95% confidence interval, 1.015-1.043) and CD4 <100 cells/microL (adjusted odds ratio, 1.524; 95% confidence interval, 1.025-2.265) were independently associated with isolated anti-HBc by logistic regression, while presence of anti-HCV and injecting drug use were not. HBV DNA was detectable in 8.3% of 277 patients with isolated anti-HBc and 14.3% of 56 patients with both anti-HBs and anti-HBc (P = 0.160). In a country hyperendemic for HBV infection, HIV-positive patients at older age and with CD4 <100 cells/microL were more likely to have isolated anti-HBc, suggesting that compromised immunity plays a role in the presence of this marker.

An HDAC inhibitor enhances the antitumor activity of a CMV promoter-driven DNA vaccine.

The cytomegalovirus (CMV) promoter is considered to be one of the strongest promoters for driving the in vivo expression of genes encoded by DNA vaccines. However, the efficacy of DNA vaccines has so far been disappointing (particularly in humans), and this might be explained in part by histone deacetylase (HDAC)-mediated chromatin condensation. Hence, we sought to investigate whether increasing the expression of DNA vaccine antigens with the HDAC inhibitor OSU-HDAC42 would enhance the efficacy of DNA vaccines in vivo. A luciferase assay was used to determine the effects of OSU-HDAC42 on CMV promoter-driven DNA plasmids in vitro and in vivo. Three HDAC inhibitors were able to activate expression from the CMV promoter in NIH3T3 cells and MBT-2 bladder cancer cells. The expression of luciferase was significantly enhanced by co-administration of pCMV-luciferase and OSU-HDAC42 in mice. To explore whether OSU-HDAC42 could enhance the specific antitumor activity of a neu DNA vaccine driven by the CMV promoter, we evaluated therapeutic effects and immune responses in a mouse tumor natively overexpressing HER2/neu. Mice receiving OSU-HDAC42 in combination with the CMV-promoter neu DNA vaccine exhibited stronger antitumor effects than mice given the DNA vaccine only. In addition, a correlation between the antitumor effects and the specific cellular immune responses was observed in the mice receiving the DNA vaccine and OSU-HDAC42.

The sodium-dependent glucose cotransporter SLC5A11 as an autoimmune modifier gene in SLE.

Genetic studies in several human autoimmune diseases suggest that the pericentromeric region of chromosome 16 might harbor an autoimmune modifier gene. We hypothesized that the sodium-dependent glucose cotransporter gene SLC5A11 is such a gene, and so might interact with immune-related genes. Herein, this hypothesis was tested in a genetic evaluation of the multiple gene effect in systemic lupus erythematosus (SLE). We used the case-control candidate gene association approach. Eight immune-related genes involved in inflammation and autoantibody generation and clear-up [interleukin 1 receptor antagonist (IL1RN), interleukin 1-beta (IL1-beta), tumor necrosis factor-alpha (TNF-alpha), lymphotoxin-alpha (LTA), tumor necrosis factor ligand superfamily, member 6 (TNFSF6), programmed cell death 1 (PDCD1), C2, and complement component 4 (C4)] were selected for study. Frequency of each candidate's genotype and allele between case and control were compared. Results were stratified by reanalyzing genotype data with relevant symptoms. Finally, improved computational data mining was used to analyze the phenotypes in a large data set. In the frequency analysis, only IL1-beta was significantly associated with SLE. Stratification analysis showed a significant association with SLE symptoms between SLC5A11 and the other immune-related genes, with the exceptions of TNFSF6 and C4. SLC5A11 was significantly associated with low C4 (as was TNF-alpha), anti-Smith antibody (anti-Sm) (as was C2), serositis, and alopecia. Finally, SLC5A11 interacted with PDCD1, TNF-alpha, LTA, and C4. After our study, we concluded that SLC5A11 is involved with some immune effects and interacts with immune-related gene(s), consistent with its function as an autoimmune modifier gene. Furthermore, SLC5A11 might induce apoptosis through the TNF-alpha, PDCD1 pathway. The present genotype-phenotype mapping approach should be applicable to genetic study of other complex diseases.

The different expression patterns of interleukin-1 receptor antagonist in systemic lupus erythematosus.

The interleukin-1 receptor antagonist (IL1RN or IL-1Ra) is a natural antagonist of IL-1-beta. Using IL1RN as a possible marker in patients with systemic lupus erythematosus (SLE), we evaluated whether uIL1RN single nucleotide polymorphisms (SNPs) were associated with the pathogenesis of SLE in Taiwanese, and specifically whether IL1RN (rs315952) was significantly associated with end-stage renal disease. We examined IL1RN isoform expression patterns in patients with SLE to determine whether the expressions play a role in the pathogenesis of SLE. Both case-control and family-based association studies were used. For the case-control study, 104 patients with SLE and 97 normal controls were recruited, and for the family-based study, 11 families with SLE without renal disorder were recruited from the 104 patients with SLE. Eight IL1RN SNPs (rs2234678, rs2234679, rs315951, rs315952, rs419598, rs432014, rs447713, and rs451578) were selected for the family-based study. Reverse-transcriptase-polymerase chain reaction (RT-PCR) was used to determine the expression pattern of each isoform. Our results showed that IL1RN (rs315952) was significantly associated with SLE in patients without renal disorder in the family-based study, after disease stratification, but was not significantly associated with SLE in the case-control study. In the family-based study, the haplotype of IL1RN (AGCCTTAG) was significantly associated with SLE (chi2 = 11.714, P < 0.001). Using RT-PCR to determine the expression pattern of the IL1RN isoforms, we found different expression patterns between normal controls and patients with SLE, with an addition of IL1RN isoform4 or the low expression of IL1RN isoform1. We concluded that IL1RN and its isoforms were involved in the pathogenesis of SLE.

Cognitive performance in cryptogenic epilepsy.

Cognitive impairment in epilepsy has begun to gain more attention in clinical practice. There is now a considerable amount of research relating to memory functioning in epilepsy, however, few studies specifically focused on cryptogenic epilepsy. We investigated the cognitive performance in cryptogenic epilepsy patients with the aid of cognitive ability screening instrument (CASI), based on cross-sectional and longitudinal aspects. A total of 100 patients who met the diagnostic criteria of cryptogenic epilepsy were recruited from a national university hospital. The patients with normal CASI scores were compared with those with abnormal ones. We also compared the follow-up CASI score after 3 years with the previous score in all cryptogenic epilepsy patients. Thirty-six per cent of cryptogenic epilepsy patients showed cognitive impairment. The variables correlated with higher risks of cognitive impairment were lower educational status, number of seizure types, duration of seizure and polytherapy, especially in the lower educational status. The correlation between CASI and the Mini-Mental State Examination was excellent. In the follow-up study, the abnormal group showed significant improvement in total CASI score. The normal group showed no significant change. We suggest that in cryptogenic epilepsy, lower educational status remains the most important factor in determining cognitive performance. Adequate treatment with antiepileptic drugs can improve cognitive performance in previously cognitively impaired patients.

Effect of ingestion of cow's milk hydrolysed formulas on whey protein-specific Th2 immune responses in naïve and sensitized mice.

BACKGROUND: For genetically predisposed atopic infants, cow's milk protein hydrolysed formulas have been widely used. OBJECTIVE: Whether hydrolysed formulas can induce oral tolerance to whey proteins will be extensively studied in naïve and sensitized mice. METHODS: Antigenicity of hydrolysed formulas was first studied using immunoblotting. Naïve mice fed hydrolysed formulas for 1-4 weeks were sensitized with whey allergens. In contrast, mice sensitized with whey allergens were fed hydrolysed formulas continually for 12 weeks. RESULTS: Whey allergens were found in Nan and Neoangelac FL. Large whey peptides with antigenicity were found in Nan-HA. Profound suppression of IgE, IgG1 and IgG responses to whey allergens were induced in those fed Nan for 1 week, or Nan-HA for 4 weeks. IgE responses to whey allergens were suppressed in those fed Neoangelac FL for 4 weeks, or Nan-HA for 1-2 weeks. In contrast, those fed extensively hydrolysed formulas for 1-4 weeks failed to show decreased responses. On the other hand, IgE responses to beta-lactoglobulin, but not to bovine serum albumin or alpha-lactalbumin, were decreased in sensitized mice fed Nan for 12 weeks. There was no suppression in sensitized mice fed hydrolysed formulas. CONCLUSION: Suppression of IgE responses to whey proteins was readily induced in naïve mice fed Nan or Nan-HA for 1 week. In contrast, it was hardly induced in sensitized mice even after prolonged feeding of Nan for 12 weeks, let alone hydrolysed formulas.

Usefulness of technetium-99m hexamethylpropylene amine oxime lung scan to detect subclinical lung injury in patients with chronic renal failure.

We used technetium-99m hexamethylpropylene amine oxime (Tc-99m HMPAO) lung scan to detect subclinical lung injury in patients with chronic renal failure (CRF), on regular hemodialysis (HD), who had normal chest X-ray findings (CXR) and pulmonary function test (PFT). The degree of pulmonary vascular endothelium damage was represented as lung/liver uptake ratios (L/L ratios) calculated by Tc-99m HMPAO lung scan. The L/L ratios of the 20 male CRF patients on HD with normal CXR and PFT were compared with those of the 20 male normal controls. The results show that the L/L ratios on Tc-99m HMPAO lung scans were significantly higher in CRF patients on HD (1.06 +/- 0.55) than those in normal controls (0.34 +/- 0.09). Using a cut-off value of 0.50, 18/20 [90%] CRF patients had abnormally increased L/L ratios. Our findings concluded that the pulmonary vascular endothelium damage represented as significantly increased L/L ratios on Tc-99m HMPAO lung scan in CRF patients on HD with normal CXR and PFT. In addition, Tc-99m HMPAO lung scan has the potential to be a sensitive, objective and noninvasive method to detect subclinical lung injury of CRF patients on HD, which is different from the traditional studies such as CXR or PFT.

Activation of Metabotropic Glutamate Receptors in Conjunction with Postsynaptic Depolarization Triggers a Long-Term Depression of the N-Methyl-D-Aspartate Receptor-Mediated Synaptic Potential in the Rat Hippocampus.

The mechanism responsible for long-term depression (LTD) of pharmacologically isolated N-methyl-D-aspartate (NMDA) receptor-mediated excitatory postsynaptic potential (EPSP(NMDA)) was studied. Intracellular recordings were made from CA1 cells of rat hippocampal slices in the presence of 6-cyano-7-nitroquinoxaline-2,3-dione (10 &mgr;M) and picrotoxin (50 &mgr;M), which block non-NMDA and GABA(A) receptors, respectively. Intracellular injections of depolarizing pulses (500 ms, 0.3-0.7 nA) at 1 Hz for 5 min in the absence of synaptic stimulation caused a persistent increase in the amplitude of EPSP(NMDA). However, coupling postsynaptic depolarization with synaptic activity induced LTD. The EPSP(NMDA) LTD could be blocked by L-2-amino-3-phosphonopropionic acid (50 &mgr;M) or (RS)-alpha-methyl-4-carboxyphenylglycine (200 &mgr;M), specific antagonists for metabotropic glutamate receptors (mGluR). Furthermore, application of trans-1-aminocyclopentane-1,3-dicarboxylic acid (t-ACPD, 50 &mgr;M), a specific mGluR agonist, in conjunction with postsynaptic depolarizing elicited LTD. In contrast, the mGluR agonists quisqualate or t-ACPD when given alone produced a sustained enhancement of EPSP(NMDA). Finally, coupled depolarization did not evoke LTD in slices pretreated with the protein kinase C (PKC) inhibitor calphostin c (60 nM). The present results demonstrate that activation of mGluR is necessary for the induction of LTD of EPSP(NMDA) and suggest that NMDA receptors are subject to bidirectional regulation by mGluR. Furthermore, the induction of LTD is likely to involve the stimulation of PKC. Copyright 1995 S. Karger AG, Basel