Clinical, electrophysiological, imaging, pathological and therapeutic observations among 18 patients with Rasmussen's encephalitis.

We studied the clinical, electrophysiological, imaging and pathological features of 18 patients with Rasmussen's encephalitis (RE). This descriptive study included 18 patients (six males, 12 females) with RE who were evaluated for demographic and phenotypic details, electroencephalogram (EEG) results, MRI results, pathological features, virological markers and outcome. Radiological staging as per Bien et al. and pathological staging in accordance with Robitaille et al. were performed. Simple partial seizures were the most common initial manifestation. During the disease course, epilepsia partialis continua (EPC) developed in 15/18 (83.3%) and hemiparesis in 17/18 (94.4%) patients. EEG revealed hemispheric slowing (100%), interictal epileptiform discharges (100%) and ictal pattern (44.4%). Brain MRI revealed unihemispheric focal cortical atrophy (100%), white matter changes (88.2%), basal ganglia-ipsilateral caudate and putamen involvement (50.0%) and progression of atrophy on serial MRI (100%). Unusual presentations in this series included late onset (n=1), and isolated lingual EPC (n=1). Diagnostic biopsies in two patients revealed Robitaille stage 3 disease. The six hemispherotomy specimens showed stage 2 disease in one, stage 3 in three and stage 4 in two cases. Heterogeneity in disease stage in the different neuroanatomical regions and within the same cortical segment reflected progression of immune-mediated damage. Immunomodulation provided only temporary benefit. Patients who underwent functional hemispherotomy had reduction in seizure frequency and improved quality of life. The clinical, EEG and MRI findings are in accordance with the established literature. MRI staging was concordant with Robitaille pathological staging. Immunomodulation did result in transient reduction in seizure frequency while surgery in six produced reasonable benefit.

Progressive multifocal leukoencephalopathy (PML) associated with HIV Clade C--is not uncommon.

Progressive multifocal leukoencephalopathy (PML) is a rare, subacute, demyelinating disease of the central nervous system caused by JC virus. Studies of PML from HIV Clade C prevalent countries are scarce. We sought to study the clinical, neuroimaging, and pathological features of PML in HIV Clade C patients from India. This is a prospective cum retrospective study, conducted in a tertiary care Neurological referral center in India from Jan 2001 to May 2012. Diagnosis was considered "definite" (confirmed by histopathology or JCV PCR in CSF) or "probable" (confirmed by MRI brain). Fifty-five patients of PML were diagnosed between January 2001 and May 2012. Complete data was available in 38 patients [mean age 39 ± 8.9 years; duration of illness-82.1 ± 74.7 days). PML was prevalent in 2.8 % of the HIV cohort seen in our Institute. Hemiparesis was the commonest symptom (44.7 %), followed by ataxia (36.8 %). Definitive diagnosis was possible in 20 cases. Eighteen remained "probable" wherein MRI revealed multifocal, symmetric lesions, hypointense on T1, and hyperintense on T2/FLAIR. Stereotactic biopsy (n = 11) revealed demyelination, enlarged oligodendrocytes with intranuclear inclusions and astrocytosis. Immunohistochemistry revelaed the presence of JC viral antigen within oligodendroglial nuclei and astrocytic cytoplasm. No differences in clinical, radiological, or pathological features were evident from PML associated with HIV Clade B. Clinical suspicion of PML was entertained in only half of the patients. Hence, a high index of suspicion is essential for diagnosis. There are no significant differences between clinical, radiological, and pathological picture of PML between Indian and Western countries.

Development and evaluation of a rapid IgM capture ELISA (JEV-Chex) for the diagnosis of Japanese encephalitis.

BACKGROUND: The IgM capture ELISA has been the most widely used diagnostic method for Japanese encephalitis. However, the lack of availability of validated commercial kits as well as the short shelf life of the kit reagents has limited the use of this technique to very few centres in Asia. OBJECTIVES: Development and evaluation of a rapid IgM capture ELISA (JEV Chex) in comparison to the conventional IgM capture ELISA. Produce key reagents such as cell culture derived JEV antigen and biotinylated monoclonal antibody which are stable at room temperature. STUDY DESIGN: The conventional IgM capture ELISA was modified to reduce the total assay time and two key reagents used in the assay JEV antigen and biotinylated anti-JEV monoclonal antibodies were rendered stable at room temperature using a special procedure. A multi-centric evaluation of this rapid ELISA was carried out using well characterized stored CSF and serum samples. Long term stability of the key reagents was also assessed over a period of 6 months. RESULTS: The rapid IgM capture ELISA developed by us showed complete concordance with the results obtained using the conventional ELISA at all the three centres where it was evaluated. In addition, the stability studies carried out with the inactivated cell culture antigen and the biotinylated monoclonal antibodies stored at room temperature for a period of 180 days revealed that both these reagents yielded consistent optical density values in the ELISA. CONCLUSIONS: The rapid ELISA format of the IgM capture ELISA (JEV-Chex) developed by us as well as the stability of reagents achieved by us in this study is what renders this rapid IgM capture ELISA very robust and user friendly since reagents can be stored at 4 degrees C by peripheral labs.

Co-existence of cerebral cysticercosis with Japanese encephalitis: a prognostic modulator.

In this study, we investigated the frequency of co-existence of cerebral cysticercosis (CC) in Japanese encephalitis (JE) cases with special emphasis on its role in predicting the final clinical outcome. Amongst the 163 confirmed cases of JE, 37.42% (61/163) had co-existent CC. This was confirmed by antibody detection in the CSF of 45 cases, CT scan of the brain in 6 cases and at autopsy in 3 cases. In 2 cases confirmation was possible by CT scan as well as at autopsy, in 4, CSF antibody levels and CT scan were suggestive of CC while in 1, CSF antibodies and autopsy were suggestive of CC The co-occurrence of Cysticercus cellulosae in the brain emerged as a prognosticator of poor outcome in JE cases (P < 0.03).

Immunoreactive antigenic sites of Cysticercus cellulosae relevant to human neurocysticercosis--immunocytochemical localization using human CSF as source of antibody.

With a view to identify the relevant antigens of Cysticercus cellulosae, recognized within human central nervous system, immunocytochemical localization of antigens on the cestode larva has been carried out using cerebrospinal fluid of patients as the source of primary antibody. CSF from nine proven cases of neurocysticercosis, two cases of culture proven tuberculous meningitis and two cases of spinal disc prolapse with no other infective or neurological disorder were used in this study. The CSF from all the cases of neurocysticercosis intensely reacted with the glycocalyx over the integument of the cyst bladder wall. The other structures recognised by the CSF antibodies were cytoplasm of the tegumentary cytons, stroma and the ductular system of the bladder wall. The cells, stroma and calcareous corpuscles of the scolex reacted variably with the CSF. Tegument of the spiral canal and sucker muscles in the scolex were not immunoreactive. We strongly suggest that the glycocalyx is the most antigenic anatomical structure of the Cysticercus cellulosae and the patients develop antibodies to it.

Cholinergic muscarinic receptors in human fetal brain: ontogeny of [3H]quinuclidinyl benzilate binding sites in corpus striatum, brainstem, and cerebellum.

The ontogeny of muscarinic receptors was studied in human fetal striatum, brainstem, and cerebellum to investigate general principles of synaptogenesis as well as the physiological balance between various chemical synapses during development in a given region of the brain. [3H]Quinuclidinyl benzilate ([3H]QNB) binding was assayed in total particulate fraction (TPF) from various parts of brain. In the corpus striatum, QNB binding sites are present at 16 weeks of gestation (average concentration 180 fmol/mg protein of TPF), slowly increase up to 24 weeks (average concentration 217 fmol/mg protein), and rapidly increase during the third trimester to 480 fmol/mg protein of TPF. In contrast, dopaminergic receptors exist as two subpopulations, one with low affinity and the other with high affinity up to the 24th week of gestation; all of them acquire the high-affinity characteristic during the third trimester. In brainstem, the muscarinic receptors show maximum concentration by 16 weeks of age (360 fmol/mg protein of TPF). Subsequently the muscarinic receptor concentration shows a gradual decline in the brainstem. In cerebellum, except for a slight increase at 24 weeks (average concentration 90 fmol/mg protein of TPF), the receptor concentration remained nearly constant at about 60-70 fmol/mg protein of TPF throughout fetal life. This study demonstrates that the ontogeny of muscarinic receptors varies among the different regions, and the patterns observed suggest that receptor formation occurs principally in the third trimester. Also noteworthy is the finding that the QNB binding sites decreased in all regions of the human brain during adult life.

Muscarinic cholinergic receptors in human foetal brain: characterization and ontogeny of [3H]quinuclidinyl benzilate binding sites in frontal cortex.

Twenty-two frontal cortices from normal human foetal brains of gestational ages ranging from 16 to 40 weeks and five postnatal brains ranging from 5 to 50 years were analysed for the ontogeny of muscarinic receptors using [3H]quinuclidinyl benzilate (QNB) as the ligand. QNB binding sites were shown to be stable up to 4 1/2 months of storage at -70 degrees C. QNB binding was characterized in frontal cortices of 28-week-old foetal brains as muscarinic receptors by the following criteria: (1) it was localised mainly in particulate fraction; (2) binding was saturable at a concentration of 1.5 nM; (3) the cholinergic antagonists atropine and scopolamine competed for the binding, with IC50 values of 1 and 0.8 nM, respectively. The agonists oxotremorine, carbachol, and pilocarpine gave IC50 values of 1, 15 and 18 microM, respectively. Nicotinic receptor ligands and noncholinergic drugs could not compete for the binding. Bimolecular association and dissociation rate constants for the reversible binding are 6.23 X 10(8) M-1 X min-1 and 2.0 X 10(-2) X min-1, respectively. The equilibrium dissociation constant is 33 pM. The KD obtained by saturation binding data is 103 pM. Ontogeny of muscarinic receptors showed three distinct phases: In phase I, they appear between 16 and 18 weeks [average concentration 109 fmol/mg protein of total particulate fraction (TPF)] and slowly increase up to 20 weeks (average concentration 147 fmol/mg protein TPF). Phase II is a lag period between 20 and 24 weeks at which time receptor concentration does not change perceptibly (average concentration (67 fmol/mg protein TPF).(ABSTRACT TRUNCATED AT 250 WORDS)