bluuebunny/crossref_metadata_2025 huggingface dataset:
Interactions of Lamotrigine with Topiramate and First‐generation Antiepileptic Drugs in the Maximal Electroshock Test in Mice: An Isobolographic Analysis
Authors: Luszczki Jarogniew J. , Czuczwar Mirosław , Kis Jacek , Krysa Jacek , Pasztelan Ireneusz , Swiader Mariusz , Czuczwar Stanislaw J.
Published: July 2003

Summary:  Purpose: The study investigated the types of interactions between lamotrigine (LTG) and first‐generation antiepileptic drugs (AEDs) or topiramate (TPM) with isobolographic analysis.

Methods: Anticonvulsant and adverse‐effect profiles of combinations of LTG with other AEDs, at fixed ratios of 1:3, 1:1, and 3:1, were evaluated in the maximal electroshock (MES)‐induced seizures and the chimney test (motor performance) in mice, which allowed the determination of benefit indices (BIs) for individual combinations.

Results: Combinations of LTG with TPM or valproate (VPA), at fixed ratios of 1:1, were significantly supraadditive (synergistic) in the MES test and, simultaneously, subadditive (antagonistic) in the chimney test, showing the best profile for AED combinations. In contrast, combinations between LTG and carbamazepine (CBZ), in terms of antiseizure protection against MES, were subadditive (antagonistic) and additive in the chimney test, resulting in unfavorable AED combinations. Moreover, the combination of LTG with phenobarbital (PB), at a fixed ratio of 1:1, despite synergy in the MES test, also was synergistic in the chimney test, resulting in a modest BI for AED combination. LTG combined with phenytoin was additive in both the MES and chimney tests in mice. The remaining combinations, at fixed ratios not mentioned earlier, also showed an average BI for AED combinations. Furthermore, LTG combined with all studied AEDs did not affect long‐term memory in mice. None of the AEDs influenced the free plasma level of LTG, whereas LTG slightly reduced the free plasma concentration of PB.

Conclusions: Interactions between LTG and TPM or LTG and VPA at a fixed ratio of 1:1 might be profitable from a preclinical point of view, displaying the most optimal BI.
Altered Hippocampal Expression of Neuropeptides in Seizure‐prone GALR1 Knockout Mice
Authors: Fetissov Sergueï O. , Jacoby Arie S. , Brumovsky Pablo R. , Shine John , Iismaa Tiina P. , Hökfelt Tomas
Published: July 2003

Summary:  Purpose: Mice carrying a deletion of the GALR1 galanin receptor have recently showed spontaneous seizure phenotype with 25% penetrance. To better understand the role of neuropeptides, which are known to undergo complex plasticity changes with development of epileptic seizures, we characterized their expression in the hippocampal formation in GALR1‐ knockout (‐KO) mice with or without seizures and in wild‐type (WT) mice.

Methods: Immunohistochemistry and in situ hybridization were used to study expression of galanin, neuropeptide Y (NPY), substance P, enkephalin, dynorphin, and cholecystokinin (CCK).

Results: In GALR1‐KO mice that had been displaying seizures, a strong upregulation of galanin immunoreactivity (ir) and messenger RNA (mRNA) was found in the polymorph layer of the dentate gyrus; galanin‐ir also appeared in a dense fiber network in the supragranular layer. A strong upregulation of enkephalin was found in the granule cells/mossy fibers, whereas dynorphin mRNA levels were modestly decreased. NPY was strongly expressed in the granule cells/mossy fibers, and an increase of NPY mRNA levels in the polymorph cells was paralleled by an increase of NPY‐ir in the molecular layer. An upregulation of substance P‐ir was confined to the fibers in the granule and molecular layers, whereas substance P mRNA was increased in the cells of the polymorph layer. Both CCK‐ir and mRNA were strongly downregulated in the granule cell/mossy fiber system, but CCK‐ir appeared increased in the supragranular and molecular layers. No changes in neuropeptide‐ir were found in GALR1‐KO mice not displaying seizures.

Conclusions: Complex changes in neuropeptide expression in some principal hippocampal neurons and interneurons appear as a characteristic feature of the spontaneous‐seizure phenotype in GALR1‐KO mice. However, to what extent causal relations exist between this “epilepsia peptidergic profile” and development of seizures requires further clarification.
Induction of Prolonged Electrographic Seizures In Vitro Has a Defined Threshold and Is All or None: Implications for Diagnosis of Status Epilepticus
Authors: Rafiq Azhar , Gong Qui‐Zhi , Lyeth Bruce G. , DeLorenzo Robert J. , Coulter Douglas A.
Published: July 2003

Summary:  Purpose: To study whether induction of prolonged (>30‐min duration) in vitro electrographic seizure discharges resembling status epilepticus (SE) is graded or all‐or‐none, and to determine the critical factors mediating SE induction.

Methods: Prolonged electrographic seizure discharges were induced in combined hippocampal–entorhinal cortical (HEC) brain slices by electrical stimulation of the Schaeffer collaterals. Discharges were recorded by using field‐potential electrodes in the dentate gyrus, CA3, CA1, and entorhinal cortex. Slices were prepared from rats that were (a) 21‐ to 30‐day‐old naive, (b) 60‐ to 120‐day old naive, (c) epileptic, and (d) status post a prior traumatic brain injury.

Results: Induction of SE discharges was dependent on the duration, but not amplitude of the preceding stimulus train–induced afterdischarge in HEC slices from 21‐ to 30‐day‐old control, brain‐injured, and epileptic animals, but not from 60‐ to 120‐day‐old animals. In slices from 21‐ to 30‐day‐old control animals, once afterdischarges exceeded 4 min in duration, SE was induced in 50% of slices, and after ≥6 min 37 s seizure activity; SE was induced in 95% of slices. A defined SE threshold also was evident in brain‐damaged rats, including rats in which an epileptic condition was induced by pilocarpine injection 4–16 weeks before recording, and rats subjected to a fluid percussive head trauma 1–8 weeks before recording. However, in these brain‐damaged animals, mean SE threshold was considerably lower (24 and 44 s, respectively). HEC slices from 60‐ to 120‐day‐old controls for the brain‐injured and epileptic animals did not develop SE even after 20 stimulations, demonstrating the pronounced effect of brain injury and epilepsy on the development of SE in the HEC slice preparation compared with that in age‐matched controls.

Conclusions: In vitro, SE discharges have a defined temporal threshold for initiation. Once a seizure exceeds 6–7 min in duration in control animals, and 30–55 s in brain‐damaged animals, the probability of SE induction is greatly increased. This demonstrates that brain injury lowers the afterdischarge duration required to produce SE and suggests that brains injured from trauma or SE are more susceptible to develop status epilepticus.
Short‐term Nonhormonal and Nonsteroid Treatment in West Syndrome
Authors: Capovilla Giuseppe , Beccaria Francesco , Montagnini Allesandra , Cusmai Raffaella , Franzoni Emilio , Moscano Filomena , Coppola Giangennero , Carotenuto Marco , Gobbi Giuseppe , Seri Stefano , Nabbout Rima , Vigevano Federico
Published: July 2003

Summary:  Purpose: West syndrome (WS) is considered an age‐dependent epileptic encephalopathy and also a particular type of electrical epileptic status. Short‐term hormonal or steroid treatment of WS with good efficacy is reported in the literature. The aim of this retrospective multiinstitutional study was to evaluate the early discontinuation of nonhormonal and nonsteroid treatment for WS.

Methods: Twenty‐two WS cases in which treatment was discontinued after a maximum of 6 months, were collected. Inclusion criteria were the presence of typical EEG hypsarrhythmia (HY) and video‐EEG recorded epileptic spasms. Exclusion criteria were the presence of partial seizures or other seizure types before spasm onset. The patients were treated with vigabatrin (VGB) in 19 cases and nitrazepam (NTZ) in three. The dose range was 70–130 mg/kg/day for VGB and 0.7–1.5 mg/kg/day for NTZ. The drug was discontinued if spasms stopped and HY disappeared after a mean treatment period of 5.1 months (range, 3–6 months). All patients underwent repeated and prolonged awake and sleep video‐EEG, both before and after drug discontinuation.

Results: Cryptogenic (15) and symptomatic (seven) WS patients were included. All the symptomatic cases had neonatal hypoxic–ischemic encephalopathy. The mean age at spasm onset was 5.5 months (range, 3–7 months; median, 6). The interval between spasm onset and drug administration ranged from 7 to 90 days (mean, 23 days; median, 20). The interval between drug administration and spasm disappearance ranged from 2 to 11 days (mean, 6 days; median, 6 days). The interval between drug administration and HY disappearance ranged from 3 to 30 days (mean, 9 days; median, 10 days). Drugs were stopped progressively over a 30‐ to 60‐day period. Follow‐up ranged from 13 to 50 months (mean, 26 months; median, 22 months). None of our cases showed spasm recurrence.

Conclusions: Our data show that successful nonhormonal and nonsteroid treatment can be shortened to a few months without spasm recurrence in patients with cryptogenic or postanoxic WS.
Identifying Potential Surgical Candidates in Patients with Evidence of Bitemporal Epilepsy
Authors: Holmes Mark D. , Miles Andrew N. , Dodrill Carl B. , Ojemann George A. , Wilensky Alan J.
Published: July 2003

Summary:  Purpose: To determine which patients with evidence of medically refractory bitemporal epilepsy are potentially good candidates for surgical therapy.

Methods: We reviewed 42 adults with intractable seizures who were found to have bitemporal ictal onsets, based on scalp video‐EEG long‐term monitoring (LTM). All underwent invasive LTM before surgery. Surgical outcomes were classified as seizure free, >75% reduction in seizures, or <75% reduction in seizures, ≥1 year after resection. We related the following factors to outcome: (a) >75% preponderance of interictal scalp EEG discharges to one temporal region; (b) magnetic resonance imaging (MRI) findings; (c) lateralizing deficits on verbal or visual reproduction memory testing; and (d) memory failure with injection contralateral to side of surgery on Wada testing.

Results: Twenty‐six (62%) of 42 patients had unilateral ictal onsets based on intracranial studies. Seizure freedom (occurring in 64% of this group), or >75% seizure reduction (found in 12% of subjects) occurred only when at least one of the following three factors was concordant with the side of surgery: preponderance of interictal scalp EEG discharges, unilateral temporal lesion on MRI, or lateralizing verbal or visual reproduction memory deficits on neuropsychological tests (p = 0.004). Seven subjects with bilateral ictal onsets based on intracranial studies had resections based on preponderance of seizures to one side, or other lateralizing noninvasive abnormality. Five of these (all of whom had ≥80% of seizures originating from one side) had >75% reduction in seizures.

Conclusions: Invasive monitoring to pursue possible surgical therapy for patients with surface EEG evidence of bitemporal epilepsy may be justified only when some lateralizing feature is found in other noninvasive assessments.
Assessment of CNS Effects of Antiepileptic Drugs by Using Quantitative EEG Measures
Authors: Salinsky Martin C. , Oken Barry S. , Storzbach Daniel , Dodrill Carl B.
Published: July 2003

Summary:  Purpose: Antiepileptic drugs (AEDs) can be associated with adverse neurologic effects including cognitive dysfunction. Objective methods for recognizing AED effects on the brain could be valuable for long‐term management. We compared quantitative EEG measures and cognitive tests in a group of patients beginning or ending AED therapy.

Methods: Subjects included 20 patients beginning AED therapy (AEDon), 12 patients stopping AED therapy (AEDoff), 33 patient controls receiving stable AED therapy (AEDco), and 73 healthy controls (Nco). All subjects underwent structured EEG recording and a cognitive test battery before change in AED dose and again 12–16 weeks later, ≥4 weeks after the last dose change. Four occipital EEG measures (peak frequency, median frequency, relative theta and delta power) were analyzed. Cognitive test changes were scored by using test–retest regression equations based on the Nco subjects. Wilcoxon tests were used for two‐group comparisons.

Results: AEDons had a significant decrease, and AEDoffs, a significant increase in the peak frequency of the EEG rhythm, as compared with controls. Results for median frequency and theta power were similar. Change in the EEG peak frequency correlated with an aggregate cognitive change measure (r2= 0.71; p < 0.001), individual cognitive measures, and subjective complaints. Of the combined AEDon/AEDoff patients, 58% exceeded the 95% confidence interval for test–retest change in EEG peak frequency.

Conclusions: Quantitative measures derived from the occipital EEG are sensitive to AEDs and correlate with AED‐related cognitive effects and subjective complaints. Although this correlation does not indicate a direct relation, quantified EEG may be a practical measure of AED impact on the brain.
Valproate Inhibits the Conversion of Testosterone to Estradiol and Acts as an Apoptotic Agent in Growing Porcine Ovarian Follicular Cells
Authors: Taubøll Erik , Gregoraszczuk Ewa L. , Kołodziej Anna , Kajta Małgorzata , Ropstad Erik
Published: July 2003

Summary:  Purpose: Long‐term valproate (VPA) treatment has been associated with hyperandrogenism and polycystic ovaries in women with epilepsy. The exact mechanisms of action of the drug on sex steroid hormone function are still unsettled. The aim of the present study was to investigate the action of VPA on basal and gonadotropin‐stimulated steroid secretion in porcine ovarian follicular cells and to measure the conversion of testosterone to estradiol. Second, the action of VPA on proliferation and apoptosis of follicular cells was investigated.

Methods: Small and medium follicles were obtained from pig ovaries on days 8–10 and 14–16 of the estrus cycle. Both follicular compartments, theca and granulosa cells, were cultured as a coculture resembling follicles in vivo. VPA in concentrations of 100 and 250 μg/ml was added to the control or gonadotropin‐stimulated cultures.

Results: VPA caused a significant increase in basal and luteinizing hormone (LH)‐stimulated testosterone secretion from small follicles, whereas in medium follicles, an increased basal but decreased LH‐stimulated testosterone secretion was found. VPA caused decreased basal and follicle‐stimulating hormone (FSH)‐stimulated estradiol secretion by small follicles, whereas only the higher concentration decreased estradiol secretion in medium follicles. The conversion of testosterone to estradiol by small follicles was decreased under the influence of VPA in testosterone‐alone and in testosterone‐plus‐FSH–stimulated cultures, whereas this was seen at only the higher VPA concentration in medium follicles. VPA had no effect on cell proliferation and viability, whereas in a dose‐dependent manner, VPA increased caspase‐3 activity.

Conclusions: VPA affected steroidogenesis in both unstimulated and gonadotropin‐stimulated porcine ovarian follicular cells and inhibited the conversion of testosterone to estradiol. In addition, VPA may act as an apoptotic agent in both small and medium‐sized follicles.
Diazepam Terminates Brief but Not Prolonged Seizures in Young, Naïve Rats
Authors: Goodkin Howard P. , Liu Xianzeng , Holmes Gregory L.
Published: July 2003

Summary:  Purpose: Ample evidence exists from both clinical and animal studies that the success of benzodiazepine intervention during status epilepticus (SE) in the mature nervous system is inversely related to seizure duration. This relationship has not been well studied in the developing nervous system.

Methods: The objective of this study was to investigate the relation of age and success of diazepam (DZP) treatment in the lithium‐pilocarpine model of secondarily generalized seizure in the rat by using naïve rats of three age groups, roughly corresponding to the human ages of infancy (P15), adolescence (P20), and adult (P60).

Results: In all age groups, the dosage of DZP that stopped the seizures at 5 min was not effective in terminating seizures at 60 min. This decline in efficacy was present as early as 15 min after seizure onset.

Conclusions: These findings demonstrate that the inverse relation between the success of benzodiazepine intervention and seizure duration is observed in young as well as in adult rats and provide further evidence that intervention for SE should commence early.
Entorhinal Cortex MRI Assessment in Temporal, Extratemporal, and Idiopathic Generalized Epilepsy
Authors: Bernasconi Neda , Andermann Frederick , Arnold Douglas L. , Bernasconi Andrea
Published: July 2003

Summary:  Purpose: We previously showed a reduction in the volume of the entorhinal cortex (EC) ipsilateral to the seizure focus in patients with intractable temporal lobe epilepsy (TLE). The purpose of this study was to examine the specificity of EC atrophy in epilepsy.

Methods: We performed volumetric measurement of the EC on high‐resolution magnetic resonance imaging (MRI) in patients with TLE (n = 70), extratemporal lobe epilepsy (ETE; n = 18), and idiopathic generalized epilepsy (IGE; n = 20). EC volumes of epilepsy patients were compared with those of 48 age‐ and sex‐matched normal controls. Within the TLE group, 63 patients were selected prospectively with hippocampal atrophy ipsilateral to the seizure focus. The remaining seven patients were chosen retrospectively based on normal volumetric MRI of the hippocampus and amygdale, as well as normal histopathologic examination of the resected tissue.

Results: Compared with normal controls, EC volume was smaller ipsilateral but not contralateral to the seizure focus in patients with TLE (p < 0.001). No difference in the EC volumes ipsilateral and contralateral to the seizure focus was seen in patients with ETE and IGE compared with normal controls. The individual analysis showed that the EC was atrophic in 73% of TLE patients with hippocampal atrophy. Three of the seven TLE patients with normal volumetric MRI of the hippocampus and amygdala and normal histopathologic examination had EC atrophy ipsilateral to the seizure focus. In no patient with ETE or IGE was the EC found to be atrophic.

Conclusions: EC atrophy ipsilateral to the seizure focus appears to be specific to mesial temporal lobe structural damage associated with TLE.
Protection Provided by Cyclosporin A against Excitotoxic Neuronal Death Is Genotype Dependent
Authors: Santos Julia Belen , Schauwecker Paula Elyse
Published: July 2003

Summary:  Purpose: Previous studies have shown that the immunosuppressant cyclosporin A (CsA), a specific blocker of the mitochondrial permeability transition (MPT) pore, can dramatically ameliorate the selective neuronal necrosis resulting from ischemia–reperfusion, traumatic brain injury, and N‐methyl‐d‐aspartate (NMDA)‐evoked neurotoxicity. The purpose of this study was to determine whether two different immunosuppressants, CsA and FK‐506, could ameliorate the neuronal damage observed after kainate‐induced seizures in strains that are differentially susceptible to excitotoxin‐induced cell death.

Methods: Excitotoxin‐resistant (C57BL/6) or ‐susceptible (FVB/N) mice were administered kainate alone (30 mg/kg), CsA alone (5, 10, or 20 mg/kg), or one of the immunosuppressants (CsA, 5 mg/kg or 10 mg/kg; FK‐506, 0.5 mg/kg) followed by kainate. After drug administration, mice were monitored continuously for the onset and extent of seizure activity. After a survival of 7 days, animals were assessed for hippocampal damage.

Results: Whereas CsA alone induced no epileptogenic effects and both immunosuppressants were without effect on the induction of kainate‐induced seizures, administration of CsA to excitotoxin‐susceptible mice (FVB/N) virtually eliminated neuronal cell death. In contrast, induction of neuronal cell death was evident when CsA was administered to excitotoxin‐resistant mice (C57BL/6). Administration of FK‐506, another commonly used immunosuppressant, which lacks an effect on the MPT, had no effect on modification of susceptibility to kainate‐induced cell death in either strain.

Conclusions: As our data show differential protection of hippocampal neurons against excitotoxic cell death by pretreatment with CsA, these results suggest that strain‐dependent differences in mitochondrial integrity and function may exist.
Angelman Syndrome: Difficulties in EEG Pattern Recognition and Possible Misinterpretations
Authors: Valente Kette D. , Andrade Joaquina Q. , Grossmann Rosi M. , Kok Fernando , Fridman Cintia , Koiffmann Célia P. , Marques‐Dias Maria J.
Published: July 2003

Summary:  Purpose: This study aimed to evaluate the sensitivity of the EEG in Angelman syndrome (AS), to verify the age at onset of suggestive EEGs and to study EEG patterns, analyzing variations and comparing our findings with nomenclature previously used.

Methods: Seventy EEG and 15 V‐EEGs of 26 patients were analyzed. Suggestive EEG patterns of AS were classified in delta pattern (DP), theta pattern (TP), and posterior discharges (PDs). Generic terms were used to simplify the analysis.

Results: Suggestive EEGs were observed in 25 (96.2%) patients. DP occurred in 22 patients with four variants—hypsarrhythmic‐like: irregular, high‐amplitude, generalized delta activity (DA) with multifocal epileptiform discharges (EDs); slow variant: regular, high‐amplitude, generalized DA with rare EDs; ill‐defined slow spike‐and‐wave: regular, high‐amplitude, generalized DA with superimposed EDs characterizing a slow wave, with notched appearance; triphasic‐like: rhythmic, moderate‐amplitude DA over anterior regions with superimposed EDs. TP was observed in eight patients, as generalized or over the posterior regions. PDs were seen in 19 patients as runs of sharp waves or runs of high‐amplitude slow waves with superimposed EDs. TP was the only age‐related pattern (younger than 8 years) and observed only in patients with deletion. In 15 patients who had an EEG before the clinical diagnosis, 60% had a suggestive tracing.

Conclusions: Although some EEG descriptions are not very detailed, and every author describes findings in a slightly different manner, obviously a common denominator must exist. In this context, EEG seems to be a very sensitive method for the diagnosis of AS, offering an opportunity to corroborate this etiologic diagnosis. Conversely, we do not believe that these patterns may be accounted as specific, except for the delta pattern, which seems to be extremely unusual in other syndromes. Other EEG patterns observed in AS, such as theta activity and PDs, occur in a wide variety of disorders. Nonetheless, their importance for the EEG diagnosis of AS is supported by the fact that they are associated with other features and may be helpful in a proper clinical setting.
Ictal Spitting: Clinical and Electroencephalographic Features
Authors: Kellinghaus Christoph , Loddenkemper Tobias , Kotagal Prakash
Published: July 2003

Summary: Purpose:To identify clinical and EEG correlates of ictal spitting automatisms and to assess their reliability in indicating the hemisphere of seizure onset.

Methods:The epilepsy‐monitoring database (1994–2002) of the Cleveland Clinic Foundation (CCF) was searched for patients with a definite history of ictal spitting. All available documents of the patients, particularly their original video and EEG data, were reviewed.

Results:Twelve (0.3%) of the ∼4,000 patients had a documented history of ictal spitting. In seven of them, 15 seizures with spitting automatisms were recorded. All of them started with an aura or arousal out of sleep. In six of the seven patients (12 of 15 seizures), EEG onset was clearly lateralized to the right, nondominant hemisphere. Spitting occurred at a median time of 21 s after EEG seizure onset. At that time, predominantly fast, high‐amplitude theta (5–7 Hz) was seen in the hemisphere of seizure onset, maximum temporal. In all but one of the total 12 patients, the epileptogenic zone was in the temporal lobe. In nine of the 12 patients, seizure onset was in the non–language‐dominant hemisphere. Two patients had seizures arising from the language‐dominant hemisphere; in another patient, the side of the seizure onset could not be determined.

Conclusions:Ictal spitting is an uncommon feature of epileptic seizures. Although the symptomatogenic area is probably outside the temporal lobe, it is most frequently seen in temporal lobe epilepsy of the right, nondominant hemisphere.
Rasmussen Syndrome: Multifocal Spread of Inflammation Suggested from MRI and PET Findings
Authors: Maeda Yumi , Oguni Hirokazu , Saitou Yoshiaki , Mutoh Ayako , Imai Kaoru , Osawa Makiko , Fukuyama Yukio , Hori Tomokatsu , Yamane Fumitaka , Kubo Osamu , Ishii Kenji , Ishiwata Kiichi
Published: July 2003

Summary:  Background: A 6‐year‐old girl with Rasmussen syndrome (RS) showed multiple small high‐signal‐intensity areas independently in the right hemisphere by fluid‐attenuated inversion recovery (FLAIR) imaging on magnetic resonance imaging (MRI) 1 year after the onset of epilepsy.

Methods: MRI performed 4 months later demonstrated a further increase in the number of these foci and enlargement in the size of the previous FLAIR lesions.

Results: An [18F]‐fluorodeoxyglucose–positron emission tomography (FDG‐PET) study showed a strong, spotty uptake in the right temporooccipital regions, corresponding to the sites of continuous EEG seizure discharges. In contrast, [11C]methionine PET demonstrated multifocal uptake regions, which corresponded anatomically to the FLAIR lesions, suggesting sites of underlying chronic inflammation.

Conclusions: These neuroimaging findings suggested that the inflammatory process in RS spreads either multifocally at the same time, as seen in this case, or from one discrete area to the adjacent region, as reported previously.
Association of the Neuronal Nicotinic Acetylcholine Receptor Subunit α4 Polymorphisms with Febrile Convulsions
Authors: Chou I‐Ching , Lee Cheng‐Chun , Huang Chao‐Ching , Wu Jer‐Yuarn , Tsai Jeffrey J. P. , Tsai Chang‐Hai , Tsai Fuu‐Jen
Published: July 2003

Summary:  Purpose: The α4‐subunit gene of the neuronal nicotinic acetylcholine receptor (CHRNA4) has been identified as the first gene underlying an idiopathic partial epilepsy syndrome in human autosomal‐dominant nocturnal frontal lobe epilepsy. Studies provided evidence that the protein coded by CHRNA4 is one of the most abundant subunits of the neuronal nicotinic acetylcholine receptors in mammalian brains, and mutations of CHRNA4 seem to cause neuronal excitation. The CHRNA4 gene may have a role in the development of febrile convulsions (FCs), the majority of childhood seizures. This study assessed the distribution of genotypes of CHRNA4 in patients with FCs.

Methods: A total of 102 children with FCs and 80 normal control subjects were included in the study. Polymerase chain reaction was used to identify the C/T polymorphism of the CHRNA4 gene. Genotypes and allelic frequencies for the CHRNA4 gene polymorphisms in both groups were compared.

Results: The number of individuals with heterozygous CHRNA4 (Ser543Ser)‐C/T genotype was significantly greater (60.8% vs. 32.5%; p = 0.001), and the CHRNA4 (Ser543Ser)‐T allele frequency was significantly higher (p = 0.001), in patients with FCs compared with healthy controls. The odds ratio for developing FCs in individuals with the CHRNA4 (Ser543Ser)‐CT genotype was 3.77 compared with individuals with two copies of the CHRNA4 (Ser543Ser)‐C allele.

Conclusions: This study demonstrated an association between the CHRNA4 gene and FCs. Individuals with the T allele had a higher incidence of FCs. These data suggest that the CHRNA4 gene or a closely linked gene might be one of the susceptibility factors for FCs.
Identification of a putative pathway for the muscle homing of stem cells in a muscular dystrophy model
Authors: Torrente Yvan , Camirand Geoffrey , Pisati Federica , Belicchi Marzia , Rossi Barbara , Colombo Fabio , El Fahime Mosthapha , Caron Nicolas J. , Issekutz Andrew C. , Constantin Gabriela , Tremblay Jacques P. , Bresolin Nereo
Published: July 2003

Attempts to repair muscle damage in Duchenne muscular dystrophy (DMD) by transplanting skeletal myoblasts directly into muscles are faced with the problem of the limited migration of these cells in the muscles. The delivery of myogenic stem cells to the sites of muscle lesions via the systemic circulation is a potential alternative approach to treat this disease. Muscle-derived stem cells (MDSCs) were obtained by a MACS® multisort method. Clones of MDSCs, which were Sca-1+/CD34−/L-selectin+, were found to adhere firmly to the endothelium of mdx dystrophic muscles after i.v. or i.m. injections. The subpopulation of Sca-1+/CD34− MDSCs expressing L-selectin was called homing MDSCs (HMDSCs). Treatment of HMDSCs with antibodies against L-selectin prevented adhesion to the muscle endothelium. Importantly, we found that vascular endothelium from striate muscle of young mdx mice expresses mucosal addressin cell adhesion molecule-1 (MAdCAM-1), a ligand for L-selectin. Our results showed for the first time that the expression of the adhesion molecule L-selectin is important for muscle homing of MDSCs. This discovery will aid in the improvement of a potential therapy for muscular dystrophy based on the systemic delivery of MDSCs.
Regulation of cell death in mitotic neural progenitor cells by asymmetric distribution of prostate apoptosis response 4 (PAR-4) and simultaneous elevation of endogenous ceramide
Authors: Bieberich Erhard , MacKinnon Sarah , Silva Jeane , Noggle Scott , Condie Brian G.
Published: July 2003

Cell death and survival of neural progenitor (NP) cells are determined by signals that are largely unknown. We have analyzed pro-apoptotic signaling in individual NP cells that have been derived from mouse embryonic stem cells. NP formation was concomitant with elevated apoptosis and increased expression of ceramide and prostate apoptosis response 4 (PAR-4). Morpholino oligonucleotide-mediated antisense knockdown of PAR-4 or inhibition of ceramide biosynthesis reduced stem cell apoptosis, whereas PAR-4 overexpression and treatment with ceramide analogs elevated apoptosis. Apoptotic cells also stained for proliferating cell nuclear antigen (a nuclear mitosis marker protein), but not for nestin (a marker for NP cells). In mitotic cells, asymmetric distribution of PAR-4 and nestin resulted in one nestin(−)/PAR-4(+) daughter cell, in which ceramide elevation induced apoptosis. The other cell was nestin(+), but PAR-4(−), and was not apoptotic. Asymmetric distribution of PAR-4 and simultaneous elevation of endogenous ceramide provides a possible mechanism underlying asymmetric differentiation and apoptosis of neuronal stem cells in the developing brain.
The ER v-SNAREs are required for GPI-anchored protein sorting from other secretory proteins upon exit from the ER
Authors: Morsomme Pierre , Prescianotto-Baschong Cristina , Riezman Howard
Published: July 2003

Glycosylphosphatidylinositol (GPI)-anchored proteins exit the ER in distinct vesicles from other secretory proteins, and this sorting event requires the Rab GTPase Ypt1p, tethering factors Uso1p, and the conserved oligomeric Golgi complex. Here we show that proper sorting depended on the vSNAREs, Bos1p, Bet1p, and Sec22p. However, the t-SNARE Sed5p was not required for protein sorting upon ER exit. Moreover, the sorting defect observed in vitro with bos1–1 extracts was also observed in vivo and was visualized by EM. Finally, transport and maturation of the GPI-anchored protein Gas1p was specifically affected in a bos1–1 mutant at semirestrictive temperature. Therefore, we propose that v-SNAREs are part of the cargo protein sorting machinery upon exit from the ER and that a correct sorting process is necessary for proper maturation of GPI-anchored proteins.
Importin β contains a COOH-terminal nucleoporin binding region important for nuclear transport
Authors: Bednenko Janna , Cingolani Gino , Gerace Larry
Published: July 2003

Proteins containing a classical NLS are transported into the nucleus by the import receptor importin β, which binds to cargoes via the adaptor importin α. The import complex is translocated through the nuclear pore complex by interactions of importin β with a series of nucleoporins. Previous studies have defined a nucleoporin binding region in the NH2-terminal half of importin β. Here we report the identification of a second nucleoporin binding region in its COOH-terminal half. Although the affinity of the COOH-terminal region for nucleoporins is dramatically weaker than that of the NH2-terminal region, sets of mutations that perturb the nucleoporin binding of either region reduce the nuclear import activity of importin β to a similar extent (∼50%). An importin β mutant with a combination of mutations in the NH2- and COOH-terminal regions is completely inactive for nuclear import. Thus, importin β possesses two nucleoporin binding sites, both of which are important for its nuclear import function.
Regulation of the expression and processing of caspase-12
Authors: Kalai Michael , Lamkanfi Mohamed , Denecker Geertrui , Boogmans Michael , Lippens Saskia , Meeus Ann , Declercq Wim , Vandenabeele Peter
Published: July 2003

Phylogenetic analysis clusters caspase-12 with the inflammatory caspases 1 and 11. We analyzed the expression of caspase-12 in mouse embryos, adult organs, and different cell types and tested the effect of interferons (IFNs) and other proinflammatory stimuli. Constitutive expression of the caspase-12 protein was restricted to certain cell types, such as epithelial cells, primary fibroblasts, and L929 fibrosarcoma cells. In fibroblasts and B16/B16 melanoma cells, caspase-12 expression is stimulated by IFN-γ but not by IFN-α or -β. The effect is increased further when IFN-γ is combined with TNF, lipopolysaccharide (LPS), or dsRNA. These stimuli also induce caspase-1 and -11 but inhibit the expression of caspase-3 and -9. In contrast to caspase-1 and -11, no caspase-12 protein was detected in macrophages in any of these treatments. Transient overexpression of full-length caspase-12 leads to proteolytic processing of the enzyme and apoptosis. Similar processing occurs in TNF-, LPS-, Fas ligand–, and thapsigargin (Tg)-induced apoptosis. However, B16/B16 melanoma cells die when treated with the ER stress–inducing agent Tg whether they express caspase-12 or not.
Infectious HIV-1 assembles in late endosomes in primary macrophages
Authors: Pelchen-Matthews Annegret , Kramer Beatrice , Marsh Mark
Published: July 2003

Although human immunodeficiency virus type 1 (HIV-1) is generally thought to assemble at the plasma membrane of infected cells, virions have been observed in intracellular compartments in macrophages. Here, we investigated virus assembly in HIV-1–infected primary human monocyte-derived macrophages (MDM). Electron microscopy of cryosections showed virus particles, identified by their morphology and positive labeling with antibodies to the viral p17, p24, and envelope proteins, in intracellular vacuoles. Immunolabeling demonstrated that these compartments contained the late endosomal marker CD63, which was enriched on vesicles within these structures and incorporated into the envelope of budding virions. The virus-containing vacuoles were also labeled with antibodies against LAMP-1, CD81, and CD82, which were also incorporated into the viral envelope. To assess the cellular source of infectious viruses derived from MDM, virus-containing media from infected cells were precipitated with specific antibodies. Only antibodies against antigens found in late endosomes precipitated infectious virus, whereas antibodies against proteins located primarily on the cell surface did not. Our data indicate that most of the infectious HIV produced by primary macrophages is assembled on late endocytic membranes and acquires antigens characteristic of this compartment. This notion has significant implications for understanding the biology of HIV and its cell–cell transmission.
Lipid rafts make for slippery platforms
Authors: Lai Eric C.
Published: July 2003

What's in a raft? Although cell membranes are certainly not homogeneous mixtures of lipids and proteins, almost all aspects of lipid rafts—how to define them, their size, composition, lifetime, and biological relevance—remain controversial. The answers will shape our views of signaling and of membrane dynamics.
P-Selectin and P-Selectin Glycoprotein Ligand 1 Are Major Determinants for Th1 Cell Recruitment to Nonlymphoid Effector Sites in the Intestinal Lamina Propria
Authors: Haddad Wael , Cooper Cristine J. , Zhang Zheng , Brown Jeffrey B. , Zhu Yuechun , Issekutz Andrew , Fuss Ivan , Lee Hae-ock , Kansas Geoffrey S. , Barrett Terrence A.
Published: July 2003

The recruitment of activated T cell subsets to sites of effector immune responses is mediated by homing receptors induced upon activation in secondary lymphoid tissue. Using an adoptive transfer model, the intestinal recruitment of CD4+ T cells activated with intraperitoneal antigen in complete Freund's adjuvant was examined. The data demonstrate that activated CD4+ T cells recruited to intestinal Peyer's patches (PP) and lamina propria (LP) up-regulate functional P-selectin glycoprotein ligand 1 (PSGL-1). Blockade of IL-12 inhibited functional PSGL-1 expression and reduced PP and LP CD4+ T cell recruitment by >40%. P-Selectin blockade reduced LP recruitment of activated cells by 56% without affecting PP recruitment. Studies of mice examined 3 d after adoptive transfer of differentiated T cell subsets revealed that Th1 but not Th2 cells were recruited to small intestine PP and LP. Mucosal addressin cell adhesion molecule blockade reduced Th1 recruitment to PP by 90% and to LP by >72%, whereas P-selectin blockade reduced Th1 recruitment to PP by 18% and Th1 recruitment to LP by 84%. These data suggest that IL-12–induced functional PSGL-1 expression is a major determinant for the recruitment of Th1 effector cells to noninflamed as well as inflamed intestine.
A Monoclonal Antibody to the α2 Domain of Murine Major Histocompatibility Complex Class I that Specifically Kills Activated Lymphocytes and Blocks Liver Damage in the Concanavalin A Hepatitis Model
Authors: Matsuoka Shuji , Tsurui Hiromichi , Abe Masaaki , Terashima Kazuo , Nakamura Kazuhiro , Hamano Yoshitomo , Ohtsuji Mareki , Honma Nakayuki , Serizawa Isao , Ishii Yasuyuki , Takiguchi Masafumi , Hirose Sachiko , Shirai Toshikazu
Published: July 2003

We earlier found that a rat monoclonal antibody (mAb) RE2 can induce rapid death of murine activated, but not resting, lymphocytes and lymphocyte cell lines, in a complement-independent manner, a cell death differing from typical apoptosis or necrosis. We here found that this cell death is independent of pathways involving Fas, caspase, and phosphoinositide-3 kinase. With the advantage of producing human B cell line transfectants with stable expression of human/mouse xeno-chimeric MHC class I genes, we found that RE2 epitope resides on the murine class I α2 domain. However, the α3 domain plays a key role in transducing the death signal, which mediates extensive aggregation of the MHC class I-integrin-actin filament system, giving rise to membrane blebs and pores. In mouse models with T/NKT cell activation-associated fulminant hepatitis, administration of mAb RE2 almost completely inhibited the development of liver cell injuries. Taken collectively, this form of cell death may be involved in homeostatic immune regulation, and induction of this form of cell death using the mAbs may be potentially therapeutic for subjects with immunological diseases mediated by activated lymphocytes.
Vav1 Phosphorylation Is Induced by β2 Integrin Engagement on Natural Killer Cells Upstream of Actin Cytoskeleton and Lipid Raft Reorganization
Authors: Riteau Béatrice , Barber Domingo F. , Long Eric O.
Published: July 2003

The guanine nucleotide exchange factor Vav1 regulates actin polymerization and contributes to cytotoxicity by natural killer (NK) cells. An open question is how Vav1 becomes activated and what receptor can signal upstream of actin cytoskeleton rearrangement upon NK cell contact with target cells. Using transfected insect cells that express ligands of human NK cell receptors, we show that engagement of the β2 integrin LFA-1 on NK cells by intercellular adhesion molecule (ICAM)-1 led to a tyrosine phosphorylation of Vav1 that was not sensitive to cholesterol depletion and to inhibition of actin polymerization. Vav1 phosphorylation was blocked by an inhibitor of Src-family kinases, and correlated with activation of its downstream effector PAK. Binding of activation receptor 2B4 to its ligand CD48 was not sufficient for Vav1 phosphorylation. However, coengagement of 2B4 with LFA-1 resulted in an enhancement of Vav1 phosphorylation that was sensitive to cholesterol depletion and to inhibition of actin polymerization. Vav1 was recruited to a detergent-resistant membrane (DRM) fraction only when 2B4 and LFA-1 were coengaged, but not after LFA-1 engagement. Therefore, binding of LFA-1 to ICAM-1 on target cells may initiate an early signaling cascade in NK cells through activation of Vav1, leading to cytoskeleton reorganization and amplification of signals from other activation receptors.
Differential Antigen Presentation Regulates the Changing Patterns of CD8+ T Cell Immunodominance in Primary and Secondary Influenza Virus Infections
Authors: Crowe Sherry R. , Turner Stephen J. , Miller Shannon C. , Roberts Alan D. , Rappolo Rachel A. , Doherty Peter C. , Ely Kenneth H. , Woodland David L.
Published: July 2003

The specificity of CD8+ T cell responses can vary dramatically between primary and secondary infections. For example, NP366–374/Db- and PA224–233/Db-specific CD8+ T cells respond in approximately equal numbers to a primary influenza virus infection in C57BL/6 mice, whereas NP366–374/Db-specific CD8+ T cells dominate the secondary response. To investigate the mechanisms underlying this changing pattern of immunodominance, we analyzed the role of antigen presentation in regulating the specificity of the T cell response. The data show that both dendritic and nondendritic cells are able to present the NP366–374/Db epitope, whereas only dendritic cells effectively present the PA224–233/Db epitope after influenza virus infection, both in vitro and in vivo. This difference in epitope expression favored the activation and expansion of NP366–374/Db-specific CD8+ memory T cells during secondary infection. The data also show that the immune response to influenza virus infection may involve T cells specific for epitopes, such as PA224–233/Db, that are poorly expressed at the site of infection. In this regard, vaccination with the PA224–233 peptide actually had a detrimental effect on the clearance of a subsequent influenza virus infection. Thus, differential antigen presentation impacts both the specificity of the T cell response and the efficacy of peptide-based vaccination strategies.
Regulation of Cyclooxygenase-2 Expression by the Translational Silencer TIA-1
Authors: Dixon Dan A. , Balch Glen C. , Kedersha Nancy , Anderson Paul , Zimmerman Guy A. , Beauchamp R. Daniel , Prescott Stephen M.
Published: July 2003

The cyclooxygenase-2 (COX-2) enzyme catalyzes the rate-limiting step of prostaglandin formation in inflammatory states, and COX-2 overexpression plays a key role in carcinogenesis. To understand the mechanisms regulating COX-2 expression, we examined its posttranscriptional regulation mediated through the AU-rich element (ARE) within the COX-2 mRNA 3′-untranslated region (3′UTR). RNA binding studies, performed to identify ARE-binding regulatory factors, demonstrated binding of the translational repressor protein TIA-1 to COX-2 mRNA. The significance of TIA-1-mediated regulation of COX-2 expression was observed in TIA-1 null fibroblasts that produced significantly more COX-2 protein than wild-type fibroblasts. However, TIA-1 deficiency did not alter COX-2 transcription or mRNA turnover. Colon cancer cells demonstrated to overexpress COX-2 through increased polysome association with COX-2 mRNA also showed defective TIA-1 binding both in vitro and in vivo. These findings implicate that TIA-1 functions as a translational silencer of COX-2 expression and support the hypothesis that dysregulated RNA-binding of TIA-1 promotes COX-2 expression in neoplasia.
Multiple T Cell Subsets Control Francisella tularensis LVS Intracellular Growth Without Stimulation Through Macrophage Interferon γ Receptors
Authors: Cowley Siobhán C. , Elkins Karen L.
Published: July 2003

A variety of data suggest that in vivo production of interferon (IFN)-γ is necessary, but not sufficient, for expression of secondary protective immunity against intracellular pathogens. To discover specific IFN-γ–independent T cell mediated mechanisms, we took advantage of an in vitro culture system that models in vivo immune responses to the intracellular bacterium Francisella tularensis live vaccine strain (LVS). LVS-immune lymphocytes specifically controlled 99% of the growth of LVS in wild-type murine bone marrow–derived macrophages. Surprisingly, LVS-immune lymphocytes also inhibited LVS intracellular growth by as much as 95% in macrophages derived from IFN-γ receptor knockout (IFNγR KO) mice. CD8+ T cells, and to a lesser degree CD4+ T cells, controlled LVS intracellular growth in both wild-type and IFNγR KO macrophages. Further, a unique population of Thy1+αβ+CD4−CD8− cells that was previously suggested to operate during secondary immunity to LVS in vivo strongly controlled LVS intracellular growth in vitro. A large proportion of the inhibition of LVS intracellular growth in IFNγR KO macrophages by all three T cell subsets could be attributed to tumor necrosis factor (TNF) α. Thus, T cell mechanisms exist that control LVS intracellular growth without acting through the IFN-γ receptor; such control is due in large part to TNF-α, and is partially mediated by a unique double negative T cell subpopulation.
The Fischer-Clifford matrices of a maximal subgroup of 𝐹𝑖′₂₄
Authors: Ali Faryad , Moori Jamshid
Published: July 2003

The Fischer group

F

i

24

′

Fi_{24}^{\prime }

is the largest sporadic simple Fischer group of order
\[

1255205709190661721292800
=

2

21

.3

16

.5
2

.7
3

.11
.13
.17
.23
.29

.

1255205709190661721292800 = 2^{21}.3^{16}.5^2.7^3.11.13.17.23.29 \;\;.

\]
The group

F

i

24

′

Fi_{24}^{\prime }

is the derived subgroup of the Fischer

3
3

-transposition group

F

i

24

Fi_{24}

discovered by Bernd Fischer. There are five classes of elements of order 3 in

F

i

24

′

Fi_{24}^{\prime }

as represented in ATLAS by

3
A

3A

,

3
B

3B

,

3
C

3C

,

3
D

3D

and

3
E

3E

. A subgroup of

F

i

24

′

Fi_{24}^{\prime }

of order

3
3

is called of type

3
X

3X

, where

X
∈
{
A
,
B
,
C
,
D
,
E
}

X \in \{A,B,C,D,E \}

, if it is generated by an element in the class

3
X

3X

. There are six classes of maximal 3-local subgroups of

F

i

24

′

Fi_{24}^{\prime }

as determined by Wilson. In this paper we determine the Fischer-Clifford matrices and conjugacy classes of one of these maximal 3-local subgroups
<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" alttext="upper G overbar colon equals upper N Subscript upper F i prime 24 Baseline left-parenthesis mathematical left-angle upper N mathematical right-angle right-parenthesis approximately-equals 3 Superscript 7 Baseline dot upper O 7 left-parenthesis 3 right-parenthesis">

G
¯

:=

N

F

i

24

′

(
⟨
N
⟩
)
≅

3
7

⋅

O
7

(
3
)

\bar {G} := N_{Fi_{24}^{\prime }}(\langle N\rangle ) \cong 3^7{\cdot }O_7(3)

, where

N
≅

3
7

N \cong 3^7

is the natural orthogonal module for

G
¯

/

N
≅

O
7

(
3
)

\bar {G}/N \cong O_7(3)

with

364
364

subgroups of type

3
B

3B

corresponding to the totally isotropic points. The group

G
¯

\bar {G}

is a nonsplit extension of

N
N

by

G
≅

O
7

(
3
)

G \cong O_7(3)

.