Correlation between Monocyte Gene Expression and Inflammation on Brain Imaging in Patients with Solitary Cerebral Cysticercus Granuloma.

Prior work has shown that 14 monocyte genes are upregulated in patients with different forms of parenchymal neurocysticercosis, including solitary cysticercus granuloma (SCG). The aim of this study was to investigate whether changes in inflammation associated with SCG seen on follow-up brain imaging are also reflected in changes in expression of these 14 genes. Peripheral blood CD14+ monocytes were isolated from 20 patients with SCG at initial diagnosis and at clinical and imaging follow-up of 6 months or more. Expressions of 14 target monocyte genes were determined by quantitative polymerase chain reaction at each visit. At a median follow-up of 14 months, the SCG had resolved in 11 patients, was persistent in four patients, and had calcified in five patients. Edema seen in the initial imaging in 17 patients had resolved in 15 patients and was markedly reduced in two patients. The expression levels of the monocyte genes LRRFIP2, TAXIBP1, and MZB1 were significantly lower at follow-up, regardless of the status of SCG on follow-up imaging. Our findings show that expression levels of monocyte genes involved with inflammatory processes decrease in patients with SCG concomitant with follow-up imaging that reveals a reduction in inflammation as revealed by complete or near-complete resolution of edema, as well as resolution or reduction in the enhancement of the granuloma.

Monocyte Gene Expression Distinguishes Enhancing Brain Parenchymal Cysticercal Granulomas From Tuberculomas.

BACKGROUND: In patients with enhancing brain parenchymal lesions, parenchymal neurocysticercosis (pNCC) is often difficult to distinguish from tuberculoma, necessitating biopsy or empirical therapy. METHODS: In a prospective study, peripheral blood monocytes were isolated from patients with definitive pNCC (n = 39) and brain tuberculomas (n = 20). Patients with tuberculomas were diagnosed by the presence of concurrent systemic tuberculosis (n = 7), pathological or bacteriological confirmation (n = 5), and resolution of typical brain lesions following a therapeutic trial of antituberculous therapy (n = 8). Expressions of 14 NCC-associated monocyte genes were determined by quantitative polymerase chain reaction and analyzed for diagnostic usefulness between the 2 groups. RESULTS: Expression of 7 genes (TAX1BP1, RAP1A, PLCG2, TOR3A, GBP1P1, LRRFIP2, and FEZ2) was significantly higher in pNCC patients than in tuberculoma patients, with TAX1BP1 and RAP1A expressions more than 22- and 5-fold higher in pNCC patients. TAX1BP1 had the highest sensitivity of 66.7% at a specificity of 100% in discriminating pNCC from tuberculoma. A combination of TAX1BP1 and RAP1A increased the sensitivity to 84.6%, and including GBP1P1 with TAX1BP1 and RAP1A further increased sensitivity to 87.2% while maintaining specificity of 100%. CONCLUSIONS: Expression of a panel of genes in blood monocytes distinguishes pNCC from brain tuberculomas in patients with enhancing brain lesions.

Distinguishing and Biochemical Phenotype Analysis of Epilepsy Patients Using a Novel Serum Profiling Platform.

Diagnosis of non-symptomatic epilepsy includes a history of two or more seizures and brain imaging to rule out structural changes like trauma, tumor, infection. Such analysis can be problematic. It is important to develop capabilities to help identify non-symptomatic epilepsy in order to better monitor and understand the condition. This understanding could lead to improved diagnostics and therapeutics. Serum mass peak profiling was performed using electrospray ionization mass spectrometry (ESI-MS). A comparison of sera mass peaks between epilepsy and control groups was performed via leave one [serum sample] out cross-validation (LOOCV). MS/MS peptide analysis was performed on serum mass peaks to compare epilepsy patient and control groups. LOOCV identified significant differences between the epilepsy patient group and control group (p = 10-22). This value became non-significant (p = 0.10) when the samples were randomly allocated between the groups and reanalyzed by LOOCV. LOOCV was thus able to distinguish a non-symptomatic epilepsy patient group from a control group based on physiological differences and underlying phenotype. MS/MS was able to identify potential peptide/protein changes involved in this epilepsy versus control comparison, with 70% of the top 100 proteins indicating overall neurologic function. Specifically, peptide/protein sera changes suggested neuro-inflammatory, seizure, ion-channel, synapse, and autoimmune pathways changing between epilepsy patients and controls.

Distinguishing patients with idiopathic epilepsy from solitary cysticercus granuloma epilepsy and biochemical phenotype assessment using a serum biomolecule profiling platform.

A major source of epilepsy is Neurocysticercosis (NCC), caused by Taenia solium infection. Solitary cysticercus granuloma (SCG), a sub-group of NCC induced epilepsy, is the most common form of NCC in India. Current diagnostic criteria for SCG epilepsy require brain imaging which may not be available in communities where the disease is endemic. Identification of serum changes and potential biomolecules that could distinguish SCG epilepsy from idiopathic generalized epilepsy (IE), without the initial need for imaging, could assist in disease identification, understanding, and treatment. The objective here was to investigate, using mass spectrometry (MS), sera biomolecule differences between patients with SCG epilepsy or IE to help distinguish these disorders based on physiological differences, to understand underlying phenotypes and mechanisms, and to lay ground work for future therapeutic and biomarker analyses. Sera were obtained from patients with SCG or IE (N = 29 each group). Serum mass peak profiling was performed with electrospray ionization (ESI) MS, and mass peak area means in the two groups were compared using leave one [serum sample] out cross validation (LOOCV). Serum LOOCV analysis identified significant differences between SCG and IE patient groups (p = 10-20), which became non-significant (p = 0.074) when the samples were randomly allocated to the groups and reanalyzed. Tandem MS/MS peptide analysis of serum mass peaks from SCG or IE patients was performed to help identify potential peptide/protein biochemical and phenotypic changes involving these two forms of epilepsy. Bioinformatic analysis of these peptide/protein changes suggested neurological, inflammatory, seizure, blood brain barrier, cognition, ion channel, cell death, and behavior related biochemical systems were being altered in these disease states. This study provides groundwork for aiding in distinguishing SCG and IE patients in minimally invasive, lower-cost manners, for improving understanding of underlying epilepsy mechanisms, and for further identifying discriminatory biomarkers and potential therapeutic targets.

Distinguishing neurocysticercosis epilepsy from epilepsy of unknown etiology using a minimal serum mass profiling platform.

Neurocysticercosis is associated with epilepsy in pig-raising communities with poor sanitation. Current internationally recognized diagnostic guidelines for neurocysticercosis rely on brain imaging, a technology that is frequently not available or not accessible in areas endemic for neurocysticercosis. Minimally invasive and low-cost aids for diagnosing neurocysticercosis epilepsy could improve treatment of neurocysticercosis. The goal of this study was to test the extent to which patients with neurocysticercosis epilepsy, epilepsy of unknown etiology, idiopathic headaches and among different types of neurocysticercosis lesions could be distinguished from each other based on serum mass profiling. For this, we collected sera from patients with neurocysticercosis-associated epilepsy, epilepsy of unknown etiology, recovered neurocysticercosis, and idiopathic headaches then performed binary group comparisons among them using electrospray ionization mass spectrometry. A leave one [serum sample] out cross validation procedure was employed to analyze spectral data. Sera from neurocysticercosis patients was distinguished from epilepsy of unknown etiology patients with a p-value of 10-28. This distinction was lost when samples were randomized to either group (p-value = 0.22). Similarly, binary comparisons of patients with neurocysticercosis who has different types of lesions showed that different forms of this disease were also distinguishable from one another. These results suggest neurocysticercosis epilepsy can be distinguished from epilepsy of unknown etiology based on biomolecular differences in sera detected by mass profiling.

Comparison of monocyte gene expression among patients with neurocysticercosis-associated epilepsy, Idiopathic Epilepsy and idiopathic headaches in India.

BACKGROUND: Neurocysticercosis (NCC), a neglected tropical disease, inflicts substantial health and economic costs on people living in endemic areas such as India. Nevertheless, accurate diagnosis using brain imaging remains poorly accessible and too costly in endemic countries. The goal of this study was to test if blood monocyte gene expression could distinguish patients with NCC-associated epilepsy, from NCC-negative imaging lesion-free patients presenting with idiopathic epilepsy or idiopathic headaches. METHODS/PRINCIPAL FINDINGS: Patients aged 18 to 51 were recruited from the Department of Neurological Sciences, Christian Medical College and Hospital, Vellore, India, between January 2013 and October 2014. mRNA from CD14+ blood monocytes was isolated from 76 patients with NCC, 10 Recovered NCC (RNCC), 29 idiopathic epilepsy and 17 idiopathic headaches patients. A preliminary microarray analysis was performed on six NCC, six idiopathic epilepsy and four idiopathic headaches patients to identify genes differentially expressed in NCC-associated epilepsy compared with other groups. This analysis identified 1411 upregulated and 733 downregulated genes in patients with NCC compared to Idiopathic Epilepsy. Fifteen genes up-regulated in NCC patients compared with other groups were selected based on possible relevance to NCC, and analyzed by qPCR in all patients' samples. Differential gene expression among patients was assessed using linear regression models. qPCR analysis of 15 selected genes showed generally higher gene expression among NCC patients, followed by RNCC, idiopathic headaches and Idiopathic Epilepsy. Gene expression was also generally higher among NCC patients with single cyst granulomas, followed by mixed lesions and single calcifications. CONCLUSIONS/SIGNIFICANCE: Expression of certain genes in blood monocytes can distinguish patients with NCC-related epilepsy from patients with active Idiopathic Epilepsy and idiopathic headaches. These findings are significant because they may lead to the development of new tools to screen for and monitor NCC patients without brain imaging.

A scalable low-cost cGMP process for clinical grade production of the HIV inhibitor 5P12-RANTES in Pichia pastoris.

In the continued absence of an effective anti-HIV vaccine, approximately 2 million new HIV infections occur every year, with over 95% of these in developing countries. Calls have been made for the development of anti-HIV drugs that can be formulated for topical use to prevent HIV transmission during sexual intercourse. Because these drugs are principally destined for use in low-resource regions, achieving production costs that are as low as possible is an absolute requirement. 5P12-RANTES, an analog of the human chemokine protein RANTES/CCL5, is a highly potent HIV entry inhibitor which acts by achieving potent blockade of the principal HIV coreceptor, CCR5. Here we describe the development and optimization of a scalable low-cost production process for 5P12-RANTES based on expression in Pichia pastoris. At pilot (150 L) scale, this cGMP compliant process yielded 30 g of clinical grade 5P12-RANTES. As well as providing sufficient material for the first stage of clinical development, this process represents an important step towards achieving production of 5P12-RANTES at a cost and scale appropriate to meet needs for topical HIV prevention worldwide.

Chronic noise stress-induced alterations of glutamate and gamma-aminobutyric acid and their metabolism in the rat brain.

Chronic stress induces neurochemical changes that include neurotransmitter imbalance in the brain. Noise is an environmental factor inducing stress. Chronic noise stress affects monoamine neurotransmitter systems in the central nervous system. The effect on other excitatory and inhibitory neurotransmitter systems is not known. The aim was to study the role of chronic noise stress on the glutamatergic and gamma-aminobutyric acid (GABA)ergic systems of the brain. Female Wistar rats (155 ± 5 g) were unintentionally exposed to noise due to construction (75-95 db, 3-4 hours/day, 5 days a week for 7-8 weeks) in the vicinity of the animal care facility. Glutamate/GABA levels and their metabolic enzymes were evaluated in different rat brain regions (cortex, hippocampus, striatum, and cerebellum) and compared with age and gender matched nonexposed rats. Chronic noise stress decreased glutamate levels and glutaminase activity 27% and 33% in the cortex, 15% and 24% in the cerebellum. Glutamate levels increased 10% in the hippocampus, 28% in striatum and glutaminase activity 15% in striatum. Glutamine synthetase activity increased significantly in all brain regions studied, that is, cortex, hippocampus, striatum, and cerebellum (P < 0.05). Noise stress-increased GABA levels and glutamate alpha decarboxylase activity 20% and 45% in the cortex, 13% and 28% in the hippocampus respectively. GABA levels and glutamate alpha decarboxylase activity decreased 15% and 14%, respectively in the striatum. GABA transaminase activity was significantly reduced in the cortex (55%), hippocampus (17%), and cerebellum (33%). Chronic noise stress differentially affected glutamatergic and GABAergic neurotransmitter systems in the rat brain, which may alter glutamate and GABA neurotransmission.

Involvement of oxidative stress, nuclear factor kappa B and the ubiquitin proteasomal pathway in dysferlinopathy.

AIMS: Dysferlinopathies are autosomal recessive neuromuscular disorders arising from mutations of the protein dysferlin that preferentially affect the limbs which waste and weaken. The pathomechanisms of the diseases are not known and effective treatment is not available. Although free radicals and upstream signaling by the redox sensitive transcription factor, NF-κB, in activation of the ubiquitin pathway are shown to occur in several muscle wasting disorders, their involvement in dysferlinopathy is not known. This study analyzed the role of oxidative stress, NF-κB and the ubiquitin pathway in dysferlinopathic muscle and in dysferlin knockdown human myoblasts and myotubes. MAIN METHODS: Fourteen dysferlinopathic muscle biopsies and 8 healthy control muscle biopsies were analyzed for oxidative stress, NF-κB activation and protein ubiquitinylation and human primary myoblasts and myotubes knocked down for dysferlin were studied for their state of oxidative stress. KEY FINDINGS: Dysferlinopathic muscle biopsies showed NF-κB p65 signaling induced protein ubiquitinylation in response to oxidative stress. Dysferlin knock down primary muscle cell cultures confirmed that oxidative stress is induced in the absence of dysferlin in muscle. SIGNIFICANCE: Anti-oxidants that also inhibit nitrosative stress and NF-κB activation, might prove to be of therapeutic benefit in slowing the progression of muscle wasting that occurs with dysferlinopathy.

Low seroprevalence of systemic cysticercosis among patients with epilepsy in Kerala--South India.

PURPOSE: Neurocysticercosis (NCC) is considered to be rare in Kerala state, India, although it is an important cause of epilepsy in many other parts of India. Our objective was to test this notion by determining the seroprevalence of cysticercosis (CC) in an unselected sample of persons with epilepsy and comparing it to that of persons without epilepsy living in Kerala. METHODS: Individuals with active epilepsy (AE) who had never resided outside Kerala state for more than one month and were attending our center for epilepsy care constituted the cases. Sex-matched persons without epilepsy who had never resided outside Kerala state for more than one month constituted the controls. The demographic details, occupation, and food habits (for the cases and controls), as well as clinical characteristics and imaging (for cases only) were recorded. Sera separated from blood drawn by venipuncture from the cases and controls were assayed for cysticercal antibodies by enzyme-linked immunoelectrotransfer blot (EITB). RESULTS: Of the 80 persons with AE, 12 were seropositive for cysticercus antibodies (15%; 95% CI: 8.8-24.4); among the 68 controls, 7 were seropositive (10.3%; 95% CI: 5.1-19.8). The odds ratio (OR) for seropositivity in the epilepsy group (1.54) was not statistically significant (95% CI: 0.6-4.2). Among the 69 patients who had a brain computed tomography (CT) scan or magnetic resonance imaging (MRI), none had features diagnostic of NCC. Gender, diet (vegetarian vs non-vegetarian, consumption of raw vegetables), drinking water status (clean vs unclean), residence (rural vs urban), exposure to manure, and animal rearing including pigs did not have any association with seropositivity. CONCLUSION: Among the residents of Kerala, most epilepsy is not related to cysticercosis.

Dimethylformamide interferes with Coomassie dye staining of proteins on blue native gel electrophoresis.

Blue native gel electrophoresis (BN-PAGE) is used extensively for characterization of mitochondrial respiratory complexes and uses the binding of Coomassie brilliant blue G-250 to visualize proteins. Oxidative modification of sulfhydryl groups of such proteins can be evaluated by labeling with iodoacetamide conjugated to biotin (BIAM) and detected with streptavidin peroxidase on Western blots following BN-PAGE. However, dissolving BIAM in dimethylformamide, a recommended solvent, reduces Coomassie blue G staining to proteins during BN-PAGE. This interference is prevented by dissolving BIAM in dimethyl sulfoxide. Precautions in the use of the dye for protein staining subsequent to BIAM labeling are discussed.

Oxidative stress, NF-κB and the ubiquitin proteasomal pathway in the pathology of calpainopathy.

The neuromuscular disorder, calpainopathy (LGMD 2A), is a major muscular dystrophy classified under limb girdle muscular dystrophies. Genetic mutations of the enzyme calpain 3 cause LGMD 2A. Calpainopathy is phenotypically observed as progressive muscle wasting and weakness. Pathomechanisms of muscle wasting of calpainopathy remain poorly understood. Oxidative stress, NF-κB and the ubiquitin proteasomal pathway underlie the pathology of several muscle wasting conditions but their role in calpainopathic dystrophy is not known. Oxidative and nitrosative stress, the source of reactive oxygen species, NF-κB signaling and protein ubiquitinylation were studied in 15 calpainopathic and 8 healthy control human muscle biopsies. Oxidative stress and NF-κB/IKK ß signaling were increased in calpainopathic muscle and may contribute to increased protein ubiquitinylation and muscle protein loss. Preventing oxidative stress or inhibition of NF-κB signaling could be considered for treatment of LGMD 2A.

The effect of acute severe monocrotophos poisoning on inhibition, expression and activity of acetylcholinesterase in different rat brain regions.

AIM: This study examined the acute effects of severe monocrotophos (MCP) poisoning on AChE inhibition, mRNA expression and recovery of acetylcholinesterase activity in different regions of the rat brain. STUDY: Wistar rats were administered monocrotophos (0.8LD(50)) by oral gavage to elicit severe effects of acute poisoning and were sacrificed 2.5 h, 24 h, 7 days, 14 days and 1 month after poisoning. Acetylcholinesterse activity, mRNA and protein were assessed in cortex, striatum, hippocampus and cerebellum. RESULTS: Acute monocrotophos administration resulted in significant AChE inhibition (50-82%) in the rat brain regions 2.5 h after poisoning. AChE inhibition was associated with down regulation of synaptic AChE mRNA 24 h after poisoning in cortex and striatum. Partial recovery of AChE activity was observed 24 h after poisoning associated with increased catalytic efficiency (K(m)) of the enzyme. The recovery of AChE mRNA and protein levels to normal occurred in 7 days in cortex and cerebellum and over one month in striatum and hippocampus. CONCLUSION: Cholinergic neurotoxicity of acute severe monocrotophos poisoning is characterized by high acetylcholinesterase inhibition, downregulation of acetylcholinesterase mRNA and slow recovery of acetylcholinesterase activity in brain regions. De novo synthesized acetylcholinesterase is associated with increased catalytic efficiency that may contribute in restoring cholinergic function.

Monocrotophos induced oxidative damage associates with severe acetylcholinesterase inhibition in rat brain.

BACKGROUND: Neurotoxicity of organophosphate pesticide poisoning, a lead cause of death in South Asia, has not been clearly elucidated. Organophosphates inhibit acetylcholinesterase and neurotoxicity is primarily a result of acetylcholine induced hyperactivation in different regions of the brain. Neurotoxicity also results from oxidative stress induced by acetylcholinesterase inhibition in the brain. Determining the severity of acetylcholinesterase inhibition that induces oxidative damage may help in developing strategies that protect the brain from organophosphate induced toxicity. AIM: To determine the level of acetylcholinesterase inhibition that induces oxidative stress in the brain following organophosphate pesticide poisoning. METHODS: Brains of rats subject to acute monocrotophos poisoning (0.8 LD(50) by gavage) were assessed for acetylcholinesterase activity, antioxidant response and oxidative damage 2.5 and 8h after poisoning and on recovery from poisoning 24h later after poisoning. Assessments were made in the cortex, striatum and hippocampus, cholinergic rich regions and cerebellum, targets of organophosphate pesticide poisoning. Analysis was in comparison to non poisoned controls. RESULTS: High acetylcholinesterase activities were noted in striatum followed by hippocampus, cerebellum and cortex. Acute severe monocrotophos poisoning inhibited acetylcholinesterase 87% in striatum, 67% in hippocampus, 58% in cerebellum, 53% in cortex and increased glutathione levels significantly in all brain regions 2.5h after poisoning. Significant lipid peroxidation and antioxidant enzymes were induced 8h after poisoning, directly correlated to high acetylcholinesterase inhibition (>67%). Recovery from monocrotophos poisoning was associated with absence of lipid peroxidation in the brain although acetylcholinesterase inhibition persisted. CONCLUSIONS: Neurotoxicity of monocrotophos poisoning is characterized by oxidative damage in regions of the brain that exhibit high acetylcholinesterase activity and severe acetylcholinesterase inhibition. Recovery from poisoning is associated with prolonged induction of antioxidants that protect against oxidative damage.

Economic implications of three strategies for the control of taeniasis.

OBJECTIVE: To evaluate the cost-effectiveness of three strategies for the control of taeniasis in a community, in terms of cost per case treated. METHODS: A study was conducted in South India to determine the prevalence of taeniasis by screening stool samples from 653 randomly chosen subjects, for coproantigens. The costs incurred in the project were used to estimate the cost per case screened and treated. A one-way sensitivity analysis was carried out for varying rates of taeniasis, different screening strategies and mass therapy. Further sensitivity analysis was carried out with different manpower and test costs. RESULTS: The rate of taeniasis as detected by ELISA for coproantigen was 3 per 1000 (2 of 653 samples). Our study showed that mass therapy without screening for taeniasis would be the most economical strategy in terms of cost per case treated if field workers are employed exclusively for either mass therapy or screening. For each strategy, costs per case treated are higher at low prevalence of taeniasis, with a sharp rise below 15%. CONCLUSIONS: In places that are endemic for taeniasis and neurocysticercosis, mass therapy or screening for taeniasis should be considered. Screening by stool microscopy is not cost-effective in terms of cost per case of taeniasis treated owing to its low sensitivity. Although the cost per case of taeniasis treated is high at low prevalence of taeniasis for all options, incorporating mass therapy into existing mass drug distribution programmes might prove to be the most cost-effective control strategy.

Clinical profile and outcome of patients hospitalized with dimethyl and diethyl organophosphate poisoning.

UNLABELLED: The two major classes of organophosphate compounds, dimethyl and diethyl organophosphates, have different toxicokinetic properties. This study evaluated the clinical profile and outcomes in patients admitted with poisoning with these two classes of organophosphates. METHODS: This retrospective study spanned 6 years (2002-2007). Patients were treated with atropine and supportive care including ventilation, as required, and followed up until death or hospital discharge. Oximes were not administered. Of the 422 charts retrieved, 396 fulfilled inclusion criteria. Data on the clinical profile, ventilation, length of hospital stay, incidence of intermediate syndrome and mortality were extracted. RESULTS: The mean (± standard deviation) age was 31.4 ± 12.7 years with a male preponderance (2.6:1). The median (interquartile range (IQR)) admission pseudocholinesterase level of 317 (222-635) U/L indicated significant inhibition of cholinesterase activity. The median lag-time to presentation to our hospital was 5 (IQR 3-8.5) hours. Oximes were administered at a primary center in 33 patients (8.3%). Dimethyl organophosphate was ingested by 141 patients, diethyl organophosphate by 108, S-alkyl organophosphate by 2, and an un-identified organophosphate by 145 patients. Ventilation was required in 260 patients (65.7%); the median duration of ventilation being 7.5 (IQR 3-12) days. Overall mortality was 13.1%. There was a significant difference between dimethyl and diethyl organophosphate compounds in ventilatory requirement (76% vs. 56%, adjusted odds ratio (OR) 2.37, 95% CI 1.01-5.57, p=0.047), duration of ventilation (11 (4-15) vs. 5 (2-9) days, adjusted OR 1.12, 95%CI 1.04-1.21, p=0.002) and incidence of intermediate syndrome (72/125 (58%) vs. 24/92 (26%), adjusted OR 2.84, 95%CI 1.38-5.86, p=0.004). Mortality was similar in the two groups (20/141 (14%) vs. 7/108 (6%), dimethyl vs. diethyl organophosphate, adjusted OR 1.29, 95%CI 0.43-3.94, p=0.65). CONCLUSIONS: Patients admitted with dimethyl organophosphate poisoning have a worse outcome compared with diethyl organophosphate poisoning for clinically relevant patient outcomes.

Neurocysticercosis, IgG immunoglobulins, and nitric oxide.

This study evaluated the role of parasite load and nitric oxide on IgG levels in neurocysticercosis. Total serum IgG, IgG antibodies specific for cysticercus antigens, and nitric oxide were compared between 85 neurocysticercosis patients, 65 with solitary cysts and 20 with multiple cysts, and 13 normal healthy controls. Sixty-six percent of patients were seropositive for cysticercus IgG antibodies. Among seropositive patients, IgG levels did not differ between those infected with multiple or solitary cysts whose serum nitric oxide levels were low (<40 nmol/ml). Among seropositive solitary cyst infected patients, IgG levels were significantly higher in those whose serum nitric oxide was low compared to those with high nitric oxide levels (p < 0.001). IgG levels were significantly higher in patients with multiple compared to single cyst infections among those negative for cysticercus antibodies (p < 0.001). Parasite load and nitric oxide modulated IgG production in neurocysticercosis. IgG levels were not determined by the number of infecting cysts in seropositive patients who did not mount a nitric oxide response. IgG production correlated to parasite load in patients negative for cysticercus antibodies.

K12-biotinylated histone H4 marks heterochromatin in human lymphoblastoma cells.

Covalent modifications of histones play crucial roles in chromatin structure and genomic stability. Recently, we reported a novel modification of histones: biotinylation of lysine residues. Here we provide evidence that K12-biotinylated histone H4 (K12Bio H4) maps specifically to both heterochromatin (alpha satellite repeats in pericentromeric regions) and transcriptionally repressed chromatin (gamma-G globin and interleukin-2) in human lymphoblastoma cells. The abundance of K12Bio H4 in these regions was similar to that of K9-dimethylated histone H3, a known marker for heterochromatin. Likewise, K8-biotinylated histone H4 (K8Bio H4) mapped to heterochromatin, but the relative enrichment was smaller compared with K12Bio H4. Stimulation of interleukin-2 transcriptional activity with phorbol-12-myristate-13-acetate and phytohemagglutinin caused a rapid depletion of K12Bio H4 in the gene promoter. These data are consistent with a novel role for biotin in chromatin structure and transcriptional activity of genes.

Conformation-sensitive immunoassays improve the serodiagnosis of solitary cysticercus granuloma in Indian patients.

Neurocysticercosis (NCC), infection of the central nervous system with larva of Taenia solium, presents in over 60% of patients in India as a solitary cysticercus granuloma (SCG). The low cyst number in these patients frequently results in an insignificant humoral response. Consequently, serological tests for patients with SCG must consider the detection of low antibody levels. Lentil lectin-specific T. solium glycoproteins of molecular weights 50, 38, 24, 18, 14 and 13 kDa are specific antigens for cysticercus antibodies in serological tests for NCC, however they do not detect antibodies in 40% of patients with SCG. To improve this rate of detection, the conformations of these protein antigens were altered to unmask additional epitopes available for antibody binding. Secondary structures of the proteins induced by reduction of disulfide bonds led to the loss of conformational epitopes necessary for cysticercus antibody recognition. Urea-induced tertiary conformations of the antigenic proteins led to the detection of antibodies in 46% of 60 patients with SCG who were serologically negative on immunoblots when the antigens were used in quaternary conformation. Conformation-sensitive immunoassays show potential for serodiagnosis of patients with SCG.

Myofibril membranes in relation to the neuromuscular weakness of acute monocrotophos poisoning.

Organophosphate poisoning is a common method of deliberate self-harm in countries where the pesticides are readily available. The severity of neuroparalysis and myopathy occurring in acute organophosphate poisoned patients is determined by the severity of poisoning and is associated with morbidity and mortality. Molecular mechanisms that underlie severe paralysis are not well delineated but are essential to know to improve treatment. In this study rats were subjected to increasing doses (0.25 to 0.8 LD(50)) of monocrotophos, and cell membrane lipid profiles, particularly those of myofibril membranes, were examined in relation to neuromuscular weakness occurring in poisoning. Increasing doses of monocrotophos inhibited brain and RBC acetylcholinesterase >/=60% early in the poisoning. RBC acetylcholinesterase levels recovered to 70% to 80% of normal while brain acetylcholinesterase remained 44% to 67% inhibited 1 week after poisoning. Increasing severity of poisoning led to significant changes in myofibril membrane lipid composition with cholesterol to phospholipid ratios increasing from 0.029 +/- 0.008 in controls to 0.063 +/- 0.023 in severe poisoning (p > 0.05). These changes were associated with neuromuscular weakness in the first day of poisoning. Membrane changes were reversible and rats recovered muscle strength in 1 week with no treatment. Lipid compositions of the intestine, brain, and muscle mitochondrial membranes were not affected by monocrotophos. The study indicated that neuromuscular weakness was associated with muscle membrane disorganization early in the course of acute organophosphate poisoning and that subclinical neurotoxicity of long duration may be a consequence of acute organophosphate poisoning. Controlling the severity of poisoning early in the course of acute organophosphate poisoning appears to be important for clinical recovery.