Central Nervous System In Order Research Articles

Rehabilitation of patients with post-traumatic stress disorder by complementary methods

Introduction. The article analyzes the relationship between stress, post-traumatic stress syndrome (PTSD) and changes in the functioning of the neuroendocrine system. Aim. Development of a hypothesis and practical evaluation of its effectiveness based on the proposed rehabilitation protocols for patients with post-traumatic stress disorder resulting from psychoemotional stress, based on a multi-level multifactorial approach with the use of transcranial stimulation by pulse currents at the frequency of rhythms of the cerebral cortex, stimulation by light pulses at the frequencies of alpha, beta and theta rhythms of the cerebral cortex in combination with neuroacoustic correction of the neuroendocrine system. Materials and methods. The study involved 36 men, combatants, aged 28-54 full years, who gave their written consent to participate in the study. In the course of the research at the first stage, a hypothesis was formed about possible options for multifactorial multilevel effects on the central nervous system in order to rehabilitate patients with PTSD. Results. The implementation of the proposed solution is to increase the efficiency and reduce the time of rehabilitation of PTSD due to a multi-level multifactorial approach: Conclusion. The results of the study revealed positive dynamics of clinical manifestations in both groups of patients. After analyzing the data obtained (assessment of the quality of life according to the SF-36 questionnaire), it was found that the protocol of rehabilitation of patients with post-traumatic stress disorder is based on the use of a multi-level approach that allows correction.

Histopathological study of neuroepithelial tumors of central nervous system

Central nervous system tumours have diverse histomorphological appearances. Accurate characterization of space-occupying lesions by histology is important for appropriate treatment. The present study is an attempt to evaluate the histomorphological patterns of the neuroepithelial tumors of central nervous system in order to provide accurate pre-therapeutic neuro-anatomic data and to facilitate better therapeutic results after surgery.To study the spectrum of neuroepithelial tumors presenting as space occupying lesions in the central nervous system.This was a prospective study carried out in the department of Pathology at Mediciti Institute of Medical Sciences over a two year period. Histopathological study of tissue specimens from neuroepithelial tumors from CNS was done. The tissue specimens were analysed for type of tumour, tumour grade and their distribution according to age, gender and topographic location. A total of 231 neoplastic space occupying lesions were studied of which neuro-epithelial tumours comprised 96 (41.5%) cases. The patient age for neruepithelial tumors ranged from 2 years to 78 years and the male to female ratio was 1.2:1. Astrocytomas were the most frequent (75%), followed by oligodendrogliomas (8.3%), oligoastrocytomas and ependymal tumours. Astrocytoma, grade IV was most common and affected the fifth decade more commonly. Astrocytomas commonly affect the frontal, temporal and parietal regions. Histopathological examination is mandatory for all space occupying lesions of CNS for correct categorization and grading of tumors. Astrocytomas are the most common of all neuroepithelial tumors. Grade IV astrocytomas most commonly affect men in fifth decade.

Central Adiponectin Signaling - A Metabolic Regulator in Support of Brain Plasticity.

Brain plasticity and metabolism are tightly connected by a constant influx of peripheral glucose to the central nervous system in order to meet the high metabolic demands imposed by neuronal activity. Metabolic disturbances highly affect neuronal plasticity, which underlies the prevalent comorbidity between metabolic disorders, cognitive impairment, and mood dysfunction. Effective pro-cognitive and neuropsychiatric interventions, therefore, should consider the metabolic aspect of brain plasticity to achieve high effectiveness. The adipocyte-secreted hormone, adiponectin, is a metabolic regulator that crosses the blood-brain barrier and modulates neuronal activity in several brain regions, where it exerts neurotrophic and neuroprotective properties. Moreover, adiponectin has been shown to improve neuronal metabolism in different animal models, including obesity, diabetes, and Alzheimer's disease. Here, we aim at linking the adiponectin's neurotrophic and neuroprotective properties with its main role as a metabolic regulator and to summarize the possible mechanisms of action on improving brain plasticity via its role in regulating the intracellular energetic activity. Such properties suggest adiponectin signaling as a potential target to counteract the central metabolic disturbances and impaired neuronal plasticity underlying many neuropsychiatric disorders.

Preliminary results of PBA-loaded nanoparticles development and the effect on oxidative stress and neuroinflammation in rats submitted to a chemically induced chronic model of MSUD.

Maple syrup urine disease (MSUD) is a genetic disorder that leads the accumulation of branched-chain amino acids (BCAA) leucine (Leu), isoleucine, valine and metabolites. The symptomatology includes psychomotor delay and mental retardation. MSUD therapy comprises a lifelong protein strict diet with low BCAA levels and is well established that high concentrations of Leu and/or its ketoacid are associated with neurological symptoms. Recently, it was demonstrated that the phenylbutyrate (PBA) have the ability to decrease BCAA concentrations. This work aimed the development of lipid-based nanoparticles loaded with PBA, capable of targeting to the central nervous system in order to verify its action mechanisms on oxidative stress and cell death in brain of rats subjected to a MSUD chronic model. PBA-loaded nanoparticles treatment was effective in significantly decreasing BCAA concentration in plasma and Leu in the cerebral cortex of MSUD animals. Furthermore, PBA modulate the activity of catalase, superoxide dismutase, glutathione peroxidase and glutathione reductase enzymes, as well as preventing the oxidative damage to lipid membranes and proteins. PBA was also able to decrease the glial fibrillary acidic protein concentrations and partially decreased the reactive species production and caspase-3 activity in MSUD rats. Taken together, the data indicate that the PBA-loaded nanoparticles could be an efficient adjuvant in the MSUD therapy, protecting against oxidative brain damage and neuroinflammation.

3158 - SCD Takes Leukemia to the CNS

Background: In acute lymphoblastic leukemia (ALL), central nervous system (CNS) involvement is a major clinical concern with no specific therapy. Prophylactic CNS-directed intrathecal chemotherapy or irradiation are required for relapse-free survival but imply long term toxicity. Therefore, mechanisms mediating leukemia-cell entry and survival into the CNS need to be understood to develop alternative CNS-directed treatment strategies. Previous studies showed an increased expression of Steroyl CoA desaturase (SCD) in BCP-ALL cells isolated from cerebrospinal fluid (CSF) of patients at CNS relapse. Aim: Investigate if SCD have a role in leukemia trafficking, homing and survival into the CNS in order to design new diagnostic and reduced toxicity- treatments to prevent relapse. Methods:Microarray profiling of cells isolated from CNS and BM of xeno-transplanted mice was performed.Human BCP-ALL cell line 018z was transduced to knock-down (KD) or overexpress human SCD. In vivo, SCD KD and overexpressing cells were transplanted in NSG mice who were sacrificed upon the first symptoms of CNS involvement. BM, spleen and meninges were collected and analyzed to check human engraftment by FACS. In Competition assays, SCD overexpressing and WT cells were transplanted in the same mouse in a 1:1 ratio. Results: Microarray profiling of cells collected from BM and meninges of leukemic mice revealed up-regulation of the genes belonging to the signaling pathway of sterol regulatory element binding proteins (SREBPs) in ALL cells that had entered the CNS, includingSCD. In vitro, SCD-KD 018z cells showed reduced proliferation than WT while SCD overexpression did not alter proliferation.Mice transplanted with SCD KD cells showed a general lower infiltration whereas SCD overexpression led to a faster onset of CNS disease.Competition assays showed that in the CNS the balance was skewed towards engraftment of SCD overexpressing cells over WT. Conclusion: SCD has a role in homing and survival of leukemic cells in the CNS.The research can lead to innovative therapies that will minimize the need for intensive chemotherapy in the CNS.

Imaging of Atypical Brain Infections and Infection Mimics in Children

AbstractAtypical brain infections in the pediatric age group are uncommon and different from the adult population; however, when present, they pose a diagnostic challenge and can result in serious and potentially fatal complications if not recognized and treated early in the course of the disease. Imaging plays a vital role because of relative inaccessibility to tissue sampling. This review article regroups the atypical pediatric brain infections (parasitic, fungal, viral, and bacterial) by age group (prenatal, perinatal, and post-neonatal period) and elucidates their characteristic imaging appearance, as well as focuses on their complications, in an aim to help clinicians (pediatricians/pediatric neuroradiologists/radiologists) better characterize, diagnose, and guide patient treatment. In addition, this review article also emphasizes on infection mimics of the central nervous system in order to empower our differential diagnosis, improve our diagnostic accuracy, and avoid unindicated patient management and treatment.

Better targeting, better efficiency of Adeno‐associated virus gene transfer in the Central Nervous System for expression of Retromer proteins

Researchers are using AAV gene transfer more and more in their neuroscience studies, including optogenetics, cre‐lox targeting, and CRISPR gene editing. Our lab has been using the AAV9 vector serotype to express genes on a wide‐scale basis throughout the central nervous system in order to study a disease that affects cells throughout the central nervous system, amyotrophic lateral sclerosis. This current project explored ways to achieve more efficient expression and in a more targeted manner than our previous work. The recently described AAV PHP.B serotype was administered to rats for the first time, and it did result in significantly more green fluorescent protein expression than did the AAV9 vector in a systematic comparison. In studies comparing the neuron‐specific synapsin promoter with our previous cytomegalovirus/ chicken β‐actin promoter, the latter promoter was noticeably more efficient yet the synapsin promoter did yield a transduction pattern with greater neuron‐selectivity, which was the goal. We also explored intracerebroventricular injections to achieve wide‐scale transduction and better avoid unwanted transduction of peripheral organs. We are combining the synapsin promoter in AAV PHP.B vectors to take advantage of these efficiency and targeting features. Our specific genes of interest that we wish to express are called the retromer proteins. Retromers make up a protein complex that is associated with the trafficking of cellular vesicles through the endosome and the trans‐Golgi network. Viral constructs were made for epitope tagged versions of the retromers VPS29 and VPS35. We hypothesize that the retromers can act to prevent the formation of beta‐amyloid plaques in Alzheimer's disease by altering the trafficking of amyloid precursor protein.Support or Funding InformationDr. Ronald Klein, Kasey Jackson, Robbie Dayton, Fidelity Biosciences Research Initiative, Karyopharm Therapeutics, Inc., ALS Association, Meira GTx, The LSU Health Sciences Center Foundation in Shreveport, The American Society for Pharmacology and Experimental Therapeutics, The Society for Toxicology

Integration of Semi-Circular Canal and Otolith Cues for Direction Discrimination during Eccentric Rotations.

Humans are capable of moving about the world in complex ways. Every time we move, our self-motion must be detected and interpreted by the central nervous system in order to make appropriate sequential movements and informed decisions. The vestibular labyrinth consists of two unique sensory organs the semi-circular canals and the otoliths that are specialized to detect rotation and translation of the head, respectively. While thresholds for pure rotational and translational self-motion are well understood surprisingly little research has investigated the relative role of each organ on thresholds for more complex motion. Eccentric (off-center) rotations during which the participant faces away from the center of rotation stimulate both organs and are thus well suited for investigating integration of rotational and translational sensory information. Ten participants completed a psychophysical direction discrimination task for pure head-centered rotations, translations and eccentric rotations with 5 different radii. Discrimination thresholds for eccentric rotations reduced with increasing radii, indicating that additional tangential accelerations (which increase with radius length) increased sensitivity. Two competing models were used to predict the eccentric thresholds based on the pure rotation and translation thresholds: one assuming that information from the two organs is integrated in an optimal fashion and another assuming that motion discrimination is solved solely by relying on the sensor which is most strongly stimulated. Our findings clearly show that information from the two organs is integrated. However the measured thresholds for 3 of the 5 eccentric rotations are even more sensitive than predictions from the optimal integration model suggesting additional non-vestibular sources of information may be involved.

Intermittent control with ankle, hip, and mixed strategies during quiet standing: A theoretical proposal based on a double inverted pendulum model

Human upright posture, as a mechanical system, is characterized by an instability of saddle type, involving both stable and unstable dynamic modes. The brain stabilizes such system by generating active joint torques, according to a time-delayed neural feedback control. What is still unsolved is a clear understanding of the control strategies and the control mechanisms that are used by the central nervous system in order to stabilize the unstable posture in a robust way while maintaining flexibility. Most studies in this direction have been limited to the single inverted pendulum model, which is useful for formalizing fundamental mechanical aspects but insufficient for addressing more general issues concerning neural control strategies. Here we consider a double inverted pendulum model in the sagittal plane with small passive viscoelasticity at the ankle and hip joints. Despite difficulties in stabilizing the double pendulum model in the presence of the large feedback delay, we show that robust and flexible stabilization of the upright posture can be established by an intermittent control mechanism that achieves the goal of stabilizing the body posture according to a “divide and conquer strategy”, which switches among different controllers in different parts of the state space of the double inverted pendulum. Remarkably, it is shown that a global, robust stability is achieved even if the individual controllers are unstable and the information exploited for switching from one controller to another is severely delayed, as it happens in biological reality. Moreover, the intermittent controller can automatically resolve coordination among multiple active torques associated with the muscle synergy, leading to the emergence of distinct temporally coordinated active torque patterns, referred to as the intermittent ankle, hip, and mixed strategies during quiet standing, depending on the passive elasticity at the hip joint.

Schwann cell expression of an oligodendrocyte-like remyelinating pattern after ethidium bromide injection in the rat spinal cord

Schwann cells are recognized by their capacity of producing single internodes of myelin around axons of the peripheral nervous system. In the ethidium bromide (EB) model of primary demyelination in the brainstem, it is observed the entry of Schwann cells into the central nervous system in order to contribute to the myelin repair performed by the oligodendrocytes that survived to the EB gliotoxic action, being able to even remyelinate more than one axon at the same time, in a pattern of repair similar to the oligodendroglial one. The present study was developed in the spinal cord to observe if Schwann cells maintained this competence of attending simultaneously different internodes. It was noted that, on the contrary of the brainstem, Schwann cells were the most important myelinogenic cells in the demyelinated site and, although rare, also presented the capacity of producing more than one internode of myelin in distinct axons.

Learning-induced lateralized activation of the MAPK/ERK cascade in identified neurons of the food-aversion network in the mollusk Helix lucorum

The MAPK/ERK pathway plays an important role in the regulation of gene expression during memory formation both in vertebrates and invertebrates. In the mollusk Helix lucorum, serotonin induces activation of MAPK/ERK in the central nervous system (CNS) upon food aversion learning. Such learning depends on a neuronal network in which specialized neurons play distinct roles so that they may exhibit different activation levels of the MAPK/ERK pathway. Here we performed a comparative analysis of MAPK/ERK activation in single neurons of the food-aversion network, focusing both on command neurons, which mediate withdrawal behavior and process information pertaining to the unconditioned stimulus, and on neurons of the procerebrum, the mollusk’s olfactory center, which process information from the conditioned stimulus. By means of Western blots designed to detect micro amounts of proteins, we determined MAPK/ERK activation in these neurons and found that after food aversion learning phospho-ERK levels increased significantly in RPa(2/3) command neurons of the right parietal ganglia and in the procerebrum. Such an increase was prevented by injection of PD98095, an inhibitor of the ERK upstream kinase (MEK-1). In contrast, no activation of MAPK/ERK was detected in similar conditions in the corresponding neurons of the left parietal ganglia LPa(2/3). This asymmetry was verified after serotonin application to the CNS in order to mimic learning. Our results thus show that learning involves synchronous and asymmetric serotonin-dependent MAPK/ERK activation. Such an asymmetry may reflect lateralization of memory processes in the mollusk brain.

Cell proliferation and death in the brain of active and hibernating frogs

'Binomial' cell proliferation and cell death have been studied in only a few non-mammalian vertebrates, such as fish. We thought it of interest to map cell proliferation/apoptosis in the brain of the frog (Rana esculenta L.) as this animal species undergoes, during the annual cycle, physiological events that could be associated with central nervous system damage. Therefore, we compared the active period and the deep underground hibernation of the frog. Using western blot analysis for proliferating cell nuclear antigen (PCNA), we revealed a positive 36 kDa band in all samples and found higher optical density values in the hibernating frogs than in active frogs. In both active and hibernating frogs, we found regional differences in PCNA-immunoreactive cells and terminal transferase dUTP nick-end labelling apoptotic cells in the ventricular zones and parenchyma areas of the main encephalon subdivisions. During the active period of the frogs, the highest concentration of PCNA-immunoreactive cells was found in the ventricle dorsal zone of the cerebral hemispheres but only some of the cells were apoptotic. By contrast, the tectal and cerebellar ventricular zones had a small or medium amount of PCNA-immunoreactive cells, respectively, and a higher number of apoptotic cells. During hibernation, an increased PCNA-immunoreactive cell number was observed in both the brain ventricles and parenchyma compared with active frogs. This increase was primarily evident in the lateral ventricles, a region known to be a proliferation 'hot spot'. Although differences existed among the brain areas, a general increase of apoptotic cell death was found in hibernating frogs, with the highest number of apoptotic cells being detected in the parenchyma of the cerebral hemispheres and optic tectum. In particular, the increased number of apoptotic cells in the hibernating frogs compared with active frogs in the parenchyma of these brain areas occurred when cell proliferation was higher in the corresponding ventricular zones. We suggest that the high number of dying cells found in the parenchymal regions of hibernating frogs might provide the stimulus for the ventricular zones to proliferate. Hibernating frogs could utilize an increased cell proliferation in the brain areas as a neuroprotective strategy to face cell death and the onset of neurological damages. Therefore, the hibernator promises to be a valuable model for studying the mechanisms naturally carried out by the central nervous system in order to adapt itself or survive adverse conditions.

Nanobiology for the Pharmacology of Cellular Ion Channels

Writing this editorial is especially pleasing. First, it provides us an opportunity to introduce new directives to the field of Neuroimmune Pharmacology and to explain why the field of nanomedicine is likely an important part of its future growth and development. Second, it is an opportunity to showcase research in this area currently operative in Russia that may not be readily accessible to the readership. Third, it is a platform to better explain why the Journal Editorial leadership was enthusiastic about the science and its relationship to the Society on NeuroImmune Pharmacology strategic goals. All are brought to bear in this issue of the Journal of Neuroimmune Pharmacology. The issue includes articles presented at a recent joint US-Russian workshop entitled, "Health in the 21st Century: Nanomedicine and Self-Organization of Biological Systems" held at M.V. Lomonosov Moscow State University (MSU), Moscow, Russia, December 10-11, 2007. The conjoint meeting was organized through the Departments of Biology, Chemistry, and Physics, MSU and by the Center for Drug Delivery and Nanomedicine and Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center (Omaha, NE). The speakers included established internationally regarded scientists from these institutions as well as graduate students and faculties at MSU. In addition to selected papers by workshop contributors, we have included several papers closely aligned to the theme of nanomedicine and nanopharmacology of the central nervous system in order to provide a biological anchor for this research. We understand that such works are new to many but hope that its organization and interdisciplinary approaches will appeal to this audience. All together, it is our hope that, by gathering basic and clinical scientists with the common interest of using nanotechnology in the delivery of therapeutic agents with a focus on nanopharmacology and complex supramolecular biological assembly, the papers included will provide a platform for thought, discussion, and future translational research.

Differential gene expression in central nervous system tissues of sheep with natural scrapie

The expression of nine genes was analyzed by real-time RT-PCR in the central nervous system in order to investigate the molecular pathogenesis of natural scrapie. An up-regulation of genes related to glial activation ( GFAP) and apoptosis ( CASP3) was detected in obex and cerebrum, indicating a reactive glia. Another glial activation-related gene ( CTSS) was slightly up-regulated in obex, whereas constitutive expression was detected for SOD1, YWHAZ, PRNP, and the apoptosis-related genes BCL2, MCL1, and BAX. This differential gene expression might reflect a spatial–temporal and tissue-specific molecular pathogenesis of scrapie.

Neuromuscular function during prolonged pedalling exercise at different cadences

The purpose of the present work was to assess the strategies set by the central nervous system in order to provide the power output required throughout a prolonged (1-h) pedalling exercise performed at different cadences (50 rpm, 110 rpm and the freely chosen cadence). Neuromuscular (NM) activity of vastus lateralis, rectus femoris, biceps femoris and gastrocnemius lateralis muscles was studied quantitatively [root-mean square (RMS) and mean power frequency (MPF)] and qualitatively (timing of onset and offset of muscle bursts during crank cycle). The present results showed that increased cadence resulted in earlier muscle activation in crank cycle. The influence of cadence on RMS and MPF depended on the considered muscle and its functional role during pedalling. Timing of onset and offset of muscle bursts was not altered by fatigue throughout the prolonged exercise. In contrast, RMS and MPF of some muscles was found to increase during prolonged exercise. In summary, the present study revealed that tonic aspects of the NM activity (RMS, MPF) are altered during prolonged pedalling exercise, while phasic aspects are remained unchanged. These results suggest that the strategies set by the central nervous system in order to provide the power output required by the exercise are held constant throughout the exercise, but that quantitative aspects of the central drive are increased in order to adapt to the progressive occurrence of the NM fatigue.

Antiviral Effects of Human Immunodeficiency Virus Type 1-Specific Small Interfering RNAs against Targets Conserved in Select Neurotropic Viral Strains

RNA interference, a natural biological phenomenon mediated by small interfering RNAs (siRNAs), has been demonstrated in recent studies to be an effective strategy against human immunodeficiency virus type 1 (HIV-1). In the present study, we used 21-bp chemically synthesized siRNA duplexes whose sequences were derived from the gp41 gene, nef, tat, and rev regions of viral RNA. These sequences are conserved in select neurotropic strains of HIV-1 (JR-FL, JR-CSF, and YU-2). The designed siRNAs exerted a potent antiviral effect on these HIV-1 strains. The antiviral effect was mediated at the RNA level (as observed by the down-regulation of the HIV-1-specific spliced transcript generating a 1.2-kbp reverse transcription [RT]-PCR product) as well as viral assembly on the cell membrane. Spliced transcripts (apart from the most abundant transcript generating a 1.2-kbp RT-PCR product) arising from an unspliced precursor likely contributed, albeit to a lesser extent, to the antiviral effect. The resultant progeny viruses had infectivities similar to that of input virus. We therefore conclude that these siRNAs interfere with the processing of the unspliced transcripts for the gp41 gene, tat, rev, and nef, eventually affecting viral assembly and leading to the overall inhibition of viral production. Apart from using the gp41 gene as a target, the conservation of each of these targets in the above-mentioned viral strains, as well as several primary isolates, would enable these siRNAs to be used as potent antiviral tools for investigations with cells derived from the central nervous system in order to evaluate their therapeutic potential and assess their utility in inhibiting HIV-1 neuropathogenesis and neuroinvasion.

Bystander CD4 T cells do not mediate demyelination in mice infected with a neurotropic coronavirus

Demyelination following infection of mice with the neurotropic coronavirus mouse hepatitis virus strain JHM (MHV) is immune-mediated. It has been demonstrated that MHV-specific CD4 and CD8 T cells are capable of causing demyelination independent of the other T cell subset. Recent work has also demonstrated that activated bystander CD8 T cells mediate significant demyelination. The ability of bystander CD4 T cells to mediate demyelination was investigated using CD4 T cell transgenic mice. The results indicated that bystander CD4 T cells were unable to cause demyelination in MHV-infected mice, despite being recruited into the central nervous system (CNS) and irrespective of activation status. These results suggest that CD4 T cells must recognize antigen in the CNS in order to cause demyelination.

On the Problem of Cooperation of the Reaction of Hands: I. Cooperation of Hand Reaction to Sound and Light Stimuli in Healthy Subjects

Coordinated reaction of hands was analyzed during examination of healthy subjects and patients with focal lesions of the central nervous system in order to find the morphological structures responsible for such cooperation. Results of investigation of coordinated hand reaction in healthy subjects to sound and light stimuli with a warning signal are presented. The degree of cooperation was estimated by the correlation coefficient between the values of the reaction time of the right and left hands under conditions of their simultaneous response. In healthy subjects, there exists a strong positive correlation, whose variability in repeated examinations is determined by constitutional features of the subject.

Immunodiagnosis of tuberculous meningitis: rapid detection of mycobacterial antigens in cerebrospinal fluid by reverse passive hemagglutination assay and their characterization by Western blotting.

Tuberculous meningitis (TBM) is one of the commonest chronic infections of the central nervous system (CNS). Diagnosis of TBM has been a problem as it causes various clinical manifestations which can be confused with those of other chronic infections of the CNS such as neurocysticercosis (NCC), neurobrucellosis and cryptococcal meningitis, that are prevalent in many underdeveloped and developing countries. Differential diagnosis of TBM can be made by detecting circulating mycobacterial antigens in CSF by immunoassays. In this study, a reverse passive hemagglutination (RPHA) has been developed using rabbit antimycobacterial IgG for detection of circulating mycobacterial antigens in CSFs from chronic infections of the CNS in order to develop a rapid, simple, sensitive and cost-effective method. Circulating mycobacterial antigens were characterized by immunoblot assay. The sensitivity limit of RPHA was 400 ng ml(-1). RPHA was specific as antimycobacterial IgG did not show any reaction with porcine Cysticercus cellulosae which was used as a control antigen. RPHA could detect mycobacterial antigens in CSF at a sensitivity level of 94.11% with a specificity of 99.0%. Immunoblot analysis of RPHA positive CSFs revealed predominantly 30-32 kDa and 71 kDa antigens whilst 6, 86, 120, 96 and 110 kDa showed varied degree of reactivity. Antigens of masses 30-32 and 71 kDa were absent in culture filtrate of Mycobacterium tuberculosis H37Rv grown in Proskeur-Beck liquid medium. RPHA is a rapid, simple and sensitive immunological method with a long shelf life of 6-8 weeks if stabilized coated erythrocytes are stored at +4 degrees C. RPHA could be used as an additional immunodiagnostic tool in both differential diagnosis and prognosis of TBM. Immunoblot results indicate that 30-32 kDa and 71 kDa antigens are cell wall derived.

Immunodiagnosis of tuberculous meningitis: rapid detection of mycobacterial antigens in cerebrospinal fluid by reverse passive hemagglutination assay and their characterization by Western blotting

Tuberculous meningitis (TBM) is one of the commonest chronic infections of the central nervous system (CNS). Diagnosis of TBM has been a problem as it causes various clinical manifestations which can be confused with those of other chronic infections of the CNS such as neurocysticercosis (NCC), neurobrucellosis and cryptococcal meningitis, that are prevalent in many underdeveloped and developing countries. Differential diagnosis of TBM can be made by detecting circulating mycobacterial antigens in CSF by immunoassays. In this study, a reverse passive hemagglutination (RPHA) has been developed using rabbit antimycobacterial IgG for detection of circulating mycobacterial antigens in CSFs from chronic infections of the CNS in order to develop a rapid, simple, sensitive and cost-effective method. Circulating mycobacterial antigens were characterized by immunoblot assay. The sensitivity limit of RPHA was 400 ng ml −1. RPHA was specific as antimycobacterial IgG did not show any reaction with porcine Cysticercus cellulosae which was used as a control antigen. RPHA could detect mycobacterial antigens in CSF at a sensitivity level of 94.11% with a specificity of 99.0%. Immunoblot analysis of RPHA positive CSFs revealed predominantly 30–32 kDa and 71 kDa antigens whilst 6, 86, 120, 96 and 110 kDa showed varied degree of reactivity. Antigens of masses 30–32 and 71 kDa were absent in culture filtrate of Mycobacterium tuberculosis H37Rv grown in Proskeur–Beck liquid medium. RPHA is a rapid, simple and sensitive immunological method with a long shelf life of 6–8 weeks if stabilized coated erythrocytes are stored at +4°C. RPHA could be used as an additional immunodiagnostic tool in both differential diagnosis and prognosis of TBM. Immunoblot results indicate that 30–32 kDa and 71 kDa antigens are cell wall derived.