Recently, the role of inflammation has attracted great attention in the pathogenesis of mesial temporal lobe epilepsy (MTLE), and microRNAs start to emerge as promising new players in MTLE pathogenesis. In this study, we investigated the dynamic expression patterns of tumor necrosis factor alpha (TNF-α) and microRNA-155 (miR-155) in the hippocampi of an immature rat model of status epilepticus (SE) and children with MTLE. The expressions of TNF-α and miR-155 were significantly upregulated in the seizure-related acute and chronic stages of MTLE in the immature rat model and also in children with MTLE. Modulation of TNF-α expression, either by stimulation using myeloid-related protein (MRP8) or lipopolysaccharide or inhibition using lenalidomide on astrocytes, leads to similar dynamic changes in miR-155 expression. Our study is the first to focus on the dynamic expression pattern of miR-155 in the immature rat of SE lithium-pilocarpine model and children with MTLE and to detect their relationship at the astrocyte level. TNF-α and miR-155, having similar expression patterns in the three stages of MTLE development, and their relationship at the astrocyte level may suggest a direct interactive relationship during MTLE development. Therefore, modulation of the TNF-α/miR-155 axis may be a novel therapeutic target for the treatment of MTLE.

: We report a case of congenital mydriasis in a neonate with megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS). Pilocarpine testing and gastrointestinal pathology in our patient suggest that the mydriasis is due to an underlying smooth muscle myopathy of the iris sphincter muscle. These findings may have important implications regarding the pathogenesis of MMIHS.

Mdivi-1 is a selective inhibitor of a mitochondrial fission protein Drp1. Recent studies demonstrated that inhibition of Drp1 provides neuroprotection in vitro and in vivo. In this study, we examined the role of mdivi-1 in hippocampal neuron death after seizures induced by pilocarpine. Our data showed that pretreatment with mdivi-1 (1.25mg/kg) significantly attenuated the neuronal death in hippocampus induced by seizures. This neuroprotective effect was dose-dependent. In addition, the seizures resulted in up-regulation of Drp1 expression and mdivi-1 treatment had no effect on the expression. Moreover, we also found that mdivi-1 (1.25mg/kg) treatment reversed the release of cytochrome c (CytC), translocation of apoptosis-inducing factor (AIF) induced by seizures while inhibiting the activated caspase-3. Altogether, our data suggested that mdivi-1 exerts neuroprotective effects against cell death of hippocampal neurons induced by seizures, and the underlying mechanism may be through inhibiting CytC release, AIF translocation and suppression of the mitochondrial apoptosis pathway.

With aim of improving the availability of drug at intraocular level and to reduce the frequency of drug administration, pilocarpine nitrate nanosuspensions were made from inert polymer resin (Eudragit RL 100) with varying drug to polymer ratios using Lutrol F68 solution in various concentration. Nanosuspensions were successfully prepared by solvent displacement method. Size of nanoparticles varied between 121.5 +/- 2.28 to 291.5 +/- 1.28 nm, a polydispersity index ranging from 0.218 +/- 0.003 to 0.658 +/- 0.035 with zeta potential ranging +14.1 +/- 0.7 to +19.8 +/- 2.3 mV. Differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy (FTIR) studies clearly suggest the compatibility of the drug with the polymer used. Scanning electron microscopy (SEM) and Transmission electron microscopy (TEM) studies completely revealed that the drug loaded nanoparticles were found to be distinct, spherical in shape having a smooth surface and the drug is molecularly dispersed uniformly throughout the whole polymer matrix. PRL4 was successfully able to sustain the drug release for 24 hr as compared to other batches of formulated nanosuspensions. No significant change in average particle size and zeta potential were observed after conducting stability studies. Results of the studies clearly suggest the suitability of Eudragit RL 100 as a promising potential drug delivery adjuvant for ocular drug administration.

The impact of regional hippocampal interactions and GABAergic transmission on ictogenesis remain unclear. Cortico-hippocampal slices from pilocarpine-treated epileptic rats were compared to controls to investigate associations between seizure-like events (SLE), GABAergic transmission, and neuronal synchrony within and between cortico-hippocampal regions. Multielectrode array recordings revealed more prevalent hippocampal SLE in epileptic tissue when excitatory transmission was enhanced and GABAergic transmission was intact (0Mg) than when GABAergic transmission was blocked (0Mg+BMI). When activity within individual regions was analyzed, spectral and temporal slow oscillation/SLE correlations and cross-correlations were highest within the hilus of epileptic tissue during SLE, but were similar in 0Mg and 0Mg+BMI. GABAergic facilitation of spectral "slow" oscillation and ripple correlations was most prominent within CA3 of epileptic tissue during SLE. When activity between regions was analyzed, slow oscillation and ripple coherence was highest between the hilus and dentate gyrus as well as between the hilus and CA3 of epileptic tissue during SLE, and was significantly higher in 0Mg than 0Mg+BMI. High 0Mg-induced SLE cross-correlations between the hilus and dentate gyrus as well as between the hilus and CA3 were reduced or abolished in 0Mg+BMI. SLE cross-correlation lag measurements provided evidence for a monosynaptic connection from the hilus to the dentate gyrus during SLE. Findings implicate the hilus as an oscillation generator, whose impact on other cortico-hippocampal regions was mediated by GABAergic transmission. Data also suggest that GABAA receptor-mediated transmission facilitated back-propagation from CA3/hilus to the dentate gyrus and that this back-propagation augmented SLE in epileptic hippocampus.

AIMS:

Salivary glands are highly susceptible to radiation and head and neck cancer patients treated with radiotherapy invariably suffer from its distressing side-effect, salivary hypofunction. The reduction in saliva disrupts oral functions, and significantly impairs oral health. Previously, we demonstrated that adenoviral-mediated expression of Tousled-like kinase 1B (TLK1B) in rat submandibular glands preserves salivary function after single dose ionizing radiation (IR). To achieve long-term transgene expression for protection of salivary gland function against fractionated radiation, this study examines the usefulness of recombinant adeno-associated viral vector for TLK1B delivery.

RESULTS:

Lactated Ringers or AAV2/9 with either TLK1B or GFP expression cassette was retroductally delivered to rat submandibular salivary glands (10e11 vg/gland), and animals were exposed, or not, to 20 Gy in 8 fractions of 2.5 Gy/day. AAV2/9 transduced predominantly the ductal cells including the convoluted granular tubules of the submandibular glands. Transgene expression after virus delivery could be detected within 5 weeks, and stable gene expression was observed till the end of study. Pilocarpine-stimulated saliva output measured at 8 weeks after completion of radiation demonstrated >10-fold reduction in salivary flow in saline- and AAV2/9-GFP- treated animals as compared to the respective non-irradiated groups (90.8 % and 92.5% reduction in salivary flow, respectively). Importantly, there was no decrease in stimulated salivary output after irradiation in animals that were pretreated with AAV2/9-TLK1B (121.5% increase in salivary flow; P<0.01). Salivary gland histology was better preserved after irradiation in TLK1B-treated group, though not significantly, compared to control groups.

CONCLUSIONS:

Single preemptive delivery of AAV2/9-TLK1B averts salivary dysfunction resulting from fractionated radiation. Although AAV2/9 transduces mostly the ductal cells of the gland, their protection against radiation assists in preserving submandibular gland function. AAV2/9-TLK1B treatment could prove beneficial in attenuating xerostomia in head and neck cancer patients undergoing radiotherapy.

Although certain metabolic characteristics such as interictal glucose hypometabolism are well established for temporal lobe epilepsy (TLE), its pathogenesis still remains unclear. Here, we performed a comprehensive study of brain metabolism in a mouse model of TLE, induced by pilocarpine-status epilepticus (SE). To investigate glucose metabolism, we injected mice 3.5-4 weeks after SE with [1,2-(13)C]glucose before microwave fixation of the head. Using (1)H and (13)C nuclear magnetic resonance spectroscopy, gas chromatography-mass spectrometry and high-pressure liquid chromatography, we quantified metabolites and (13)C labeling in extracts of cortex and hippocampal formation (HF). Hippocampal levels of glutamate, glutathione and alanine were decreased in pilocarpine-SE mice compared with controls. Moreover, the contents of N-acetyl aspartate, succinate and reduced nicotinamide adenine dinucleotide (phosphate) NAD(P)H were decreased in HF indicating impairment of mitochondrial function. In addition, the reduction in (13)C enrichment of hippocampal citrate and malate suggests decreased tricarboxylic acid (TCA) cycle turnover in this region. In cortex, we found reduced (13)C labeling of glutamate, glutamine and aspartate via the pyruvate carboxylation and pyruvate dehydrogenation pathways, suggesting slower turnover of these amino acids and/or the TCA cycle. In conclusion, mitochondrial metabolic dysfunction and altered amino-acid metabolism is found in both cortex and HF in this epilepsy model.Journal of Cerebral Blood Flow & Metabolism advance online publication, 24 April 2013; doi:10.1038/jcbfm.2013.54.

The aim of this study was to investigate the effect of Withania somnifera (WS) extract, withanolide A (WA), and carbamazepine (CBZ) on cerebellar AMPA receptor function in pilocarpine-induced temporal lobe epilepsy (TLE). In the present study, motor learning deficit was studied by rotarod test, grid walk test, and narrow beam test. Motor learning was significantly impaired in rats with epilepsy. The treatment with WS and WA significantly reversed the motor learning deficit in rats with epilepsy when compared with control rats. There was an increase in glutamate content and IP3 content observed in rats with epilepsy which was reversed in WS- and WA-treated rats with epilepsy. alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor dysfunction was analyzed using radiolabeled AMPA receptor binding assay, AMPA receptor mRNA expression, and immunohistochemistry using anti-AMPA receptor antibody. Our results suggest that there was a decrease in Bmax, mRNA expression, and AMPA receptor expression indicating AMPA receptor dysfunction, which is suggested to have contributed to the motor learning deficit observed in rats with epilepsy. Moreover, treatment with WS and WA resulted in physiological expression of AMPA receptors. There was also alteration in GAD and GLAST expression which supplemented the increase in extracellular glutamate. The treatment with WS and WA reversed the GAD and GLAST expression. These findings suggest that WS and WA regulate AMPA receptor function in the cerebellum of rats with TLE, which has therapeutic application in epilepsy.

A 64-year-old man with Marfan's syndrome was referred with ectopia lentis and visual acuity (VA) of 6/12 bilaterally. He had a history of right BRVO, aortic aneurysm stenting and multiple TIAs and was on warfarin and clopidogrel. He was listed for a right vitreolensectomy. Overnight, however, he had severe left eye pain and vomiting that gradually resolved. On examination, his VA was 6/60 and his pupil was mid-dilated. The cornea was oedematous (figure 1) but his intraocular pressure (IOP) was 14mmHg. He was given pilocarpine drops and listed for a left vitreolensectomy. However, he presented again with an IOP of 48mmHg and a pupil-captured crystalline lens (figure 2). His IOP was successfully controlled medically with intravenous Mannitol, acetazolamide and topical hypotensives, and an Nd-YAG iridotomy was arranged to prevent further episodes. However, upon corneal indentation during gonioscopy the lens relocated to the posterior chamber. The iridotomy was completed and the patient suffered no further acute angle closure episodes. He underwent vitreolensectomy and anterior chamber lens implantation in both eyes. Postoperatively, his best corrected VA was 6/6 and IOP was 18 mmHg.

To determine the efficacy and compare the side-effects of cevimeline and pilocarpine in the secretion of saliva in patients with xerostomia.A randomized, cross-over, double blind study was designed. Fifteen patients with diagnosis of xerostomia were assigned to take either 5 mg of pilocarpine or 30 mg of cevimeline three times a day for four weeks. Salivary flow rates were measured during the initial baseline, first and second month appointments. Statistical analysis was carried out with ANOVA and post hoc t-tests.Twelve patients completed both medication treatments. Although both medications proved to increase salivary secretion, there was no significant difference between pilocarpine and cevimeline. Also, the perceived side-effects between the two medications were similar.Both medications increased the secretion of saliva at the end of four weeks. However, there was a slightly higher increment in saliva with pilocarpine. However, the difference was not statistically significant.