An efficient Gait Dynamics classification method for Neurodegenerative Diseases using Brain signals.

Neurons of the human brain are primarily affected by the Huntington's disease (HD), Amyotrophic Lateral Sclerosis (ALS), Parkinson's disease and so on. Classification of these neurodegenerative diseases (NDD) is clinically important to analyze the destruction of nerve cells. Early diagnosis of NDD'S helps in saving the human life. Based on the report of previous studies, motor impairment or human gait cycle is largely affected by the clinical symptoms of NDD. Accurate diagnosis of various neurodegenerative diseases in correct time is very important for early diagnosis of the disease. Diseases can be diagnosed earlier by means of characterizing the gait cycle. In this work, a gait dynamics classification method is proposed for determining the neurodegenerative diseases from the brain signals using multilevel feature extraction method. From force sensitive resistors, the left and right feet signals recorded in 60 one minute are included in the input database. It is obtained through fixing 16 healthy subjects, 13 ALS, 20 HD, and 15 PD. Using six levels of Discrete Wavelet Transform (DWT), the features are determined by means of decomposing the raw signal. Ultimately, the pathological gait signals are classified through exploiting three multilevel feature extraction techniques named as, (Detrended Fluctuation Analysis (DFA), Positive, Negative Peak Histogram Analysis (PNPHA) (proposed Method) and Statistical Temporal parameter Analysis (STA)). Experimental outcomes proved that the gait dynamics are successively distinguished between NDD and group of healthy controls using the proposed method.

The neuronal ceroid lipofuscinoses in human EPMR and mnd mutant mice are associated with mutations in CLN8.

The neuronal ceroid lipofuscinoses (NCLs) are a genetically heterogeneous group of progressive neurodegenerative disorders characterized by the accumulation of autofluorescent lipopigment in various tissues. Progressive epilepsy with mental retardation (EPMR, MIM 600143) was recently recognized as a new NCL subtype (CLN8). It is an autosomal recessive disorder characterized by onset of generalized seizures between 5 and 10 years, and subsequent progressive mental retardation. Here we report the positional cloning of a novel gene, CLN8, which is mutated in EPMR. It encodes a putative transmembrane protein. EPMR patients were homozygous for a missense mutation (70C-->G, R24G) that was not found in homozygosity in 433 controls. We also cloned the mouse Cln8 sequence. It displays 82% nucleotide identity with CLN8, conservation of the codon harbouring the human mutation and is localized to the same region as the motor neuron degeneration mouse, mnd, a naturally occurring mouse NCL (ref. 4). In mnd/mnd mice, we identified a homozygous 1-bp insertion (267-268insC, codon 90) predicting a frameshift and a truncated protein. Our data demonstrate that mutations in these orthologous genes underlie NCL phenotypes in human and mouse, and represent the first description of the molecular basis of a naturally occurring animal model for NCL.

[HIV and risk factors for the blood donors at the central hospital of Yaounde, Cameroon]. / VIH et facteurs de risque chez les donneurs de sang de remplacement familiaux et les bénévoles à l'hôpital central de Yaoundé, Cameroun.

The HIV/AIDS infection is in a permanent progress in Cameroon. Through this descriptive and analytical cross-sectional study, we aimed to compare the occurrence of the HIV by taking into account the risks factors that are significantly associated with HIV. The investigation was carried out from 1 January till 31 December 2009 in the Blood Bank of the Central Hospital of Yaounde in Cameroon. A structured questionnaire was proposed to collect socio-demographic and risk behavioral information. Venous blood was collected for HIV antibody testing. Generalized estimating equation with logistic regression was used to analyze the risk factors for HIV infection. In all, 5 058 persons were included in this study. Serological examination revealed a total prevalence of 5.4% of HIV infection in the population studied. The family/replacement donors constituted the majority (69.5%) and showed a higher risk of seropositivity of HIV than the benevolent donors in raw analysis; but after adjustment, the family donors had the same risk of seropositivity of HIV than voluntary blood donors (aOR = 1.00). Variables such as homosexual intercourse (aOR = 1.61), to have already made a screening test of HIV (aOR = 1.83), mobility (aOR = 2.24), treatment and records of STI (aOR = 3.81), use of the condom (aOR = 6.63), more than one sexual partner (aOR = 8.40) remained significantly linked to the result of the HIV serology and constituted risk factors that will be emphasized during the selection of the donors.

A model for Batten disease protein CLN3: functional implications from homology and mutations.

In an attempt to understand the molecular nature of Batten disease, we have examined the amino acid sequence of the affected CLN3 gene product (The International Batten Disease Consortium (1995) Cell 82, 949-957) and the site-specific mutations which give rise to the biological defect. Homology searches and molecular modeling have led to the development of a model for the folding and disposition of the protein, possibly within a mitochondrial membrane. High homology with a yeast protein of unknown function suggests a strong evolutionary conservation of function. We speculate that a possible role for the protein may be in chaperoning the folding/unfolding or assembly/ disassembly of other proteins, specifically subunit c of the mitochondrial ATP synthase complex.

Delayed classic and protracted phenotypes of compound heterozygous juvenile neuronal ceroid lipofuscinosis.

OBJECTIVE: To correlate the phenotypes with the genotypes of 10 Finnish juvenile neuronal ceroid lipofuscinosis (JNCL; late-onset Batten disease) patients who all are compound heterozygotes for the major 1.02-kb deletion in the CLN3 gene. METHODS: The mutations on the non-1.02-kb deletion chromosomes were screened in 6 patients; in the other 4 patients the mutations were known (one affecting a splice site, two missense mutations, and one deletion of exons 10 through 13). Clinical features were examined, and MRI, MRS, somatosensory evoked magnetic field (SEF), and overnight polysomnography (PSG) studies were performed. RESULTS: A novel deletion of exons 10 through 13 was found in 6 patients belonging to three families. In the patients carrying the deletions of exons 10 through 13 the clinical course of the disease was fairly similar. Variation was greatest in the time course to blindness. In these patients the mental and motor decline was slower than in classic JNCL, but more severe than in the two patients with missense mutations in exons 11 and 13. MRI showed brain atrophy in 4 patients. One patient had hyperintense periventricular white matter, otherwise brain signal intensities were normal. SEFs were enhanced in patients older than 14 years, whereas in PSG all but the youngest 6-year-old patient showed epileptiform activity in slow-wave sleep. CONCLUSIONS: JNCL can manifest as at least three different phenotypes: classic, delayed classic, and protracted JNCL with predominantly ocular symptoms. Finnish compound heterozygotes have the delayed classic or the protracted form of JNCL.

Analysis of Batten disease candidate genes STP and STM.

We have sequenced a large proportion of the open reading frames (ORFs) of two phenol sulphotransferase gene transcripts (STP and STM) from three patients with Batten disease. This was done using reverse transcription and PCR amplification of total RNA followed by direct sequencing of the PCR products. No mutations or changes have been observed in either gene after sequencing 93% of the STP ORF and 72% of the STM ORF. Work is in progress to finish sequencing both genes which will allow the confirmation or exclusion of these phenol sulphotransferases having a role in the development of Batten disease.

Physical map of the region containing the gene for Batten disease (CLN3).

CLN3 has been mapped genetically to 16p12, to the interval between D16S288 and D16S383, a sex-averaged genetic distance of 2.1 cM. Analysis of disease haplotypes for four microsatellite markers in this interval, D16S288, D16S299, D16S298, and SPN, has shown significant allelic association between one allele at each of these loci and CLN3. All four of the associated markers were used as nucleation sites in the isolation of genomic clones (YACs). A contig was assembled which contains 3 of the 4 associated markers and which confirmed the relative order of these markers. Marker D16S272 has been located on the physical map between D16S288 and D16S299. Restriction mapping has demonstrated the location of possible CpG islands. One gene, STP, has been localised on the YAC contig proximal to D16S298 and is therefore a candidate for CLN3. Other genes, including IL4R, SGLT2, and UQCRC2, have been excluded from this region.

Phenol sulfotransferases: candidate genes for Batten disease.

Batten disease (juvenile-onset neuronal ceroid lipofuscinosis; JNCL) is an autosomal recessive neurodegenerative disorder, characterized by the cytosomal accumulation of autofluorescent proteolipopigments in neurons and other cell types. The Batten disease gene (CLN3) has not yet been identified, but has been mapped to a small region of human chromosome area 16p12.1-p11.2. We recently reported the fortuitous discovery that the cytosolic phenol sulfotransferase gene (STP) is located within this same interval of chromosome 16p. Since phenol sulfotransferase is expressed in neurons, can sulfate lipophilic phenolic compounds, and is mapped near CLN3, STP is considered as a candidate gene for Batten disease. YAC and cosmid cloning results have further substantiated the close proximity of STP and a highly related sulfotransferase (STM), encoding the catecholamine-preferring enzyme, to the CLN3 region of chromosome 16p. In this report, we summarize some of the recent progress in the identification of two phenol sulfotransferase genes (STP and STM) as positional candidate genes for Batten disease.

Role of Tannins in Defending Plants Against Ruminants: Reduction in Dry Matter Digestion?

Polyphenolic allelochemicals, such as tannins, are widely thought to reduce the digestibility of plants consumed by herbivores by binding to digestive enzymes and dietary proteins. While the apparent digestibility of protein and, therefore, cell solubles is reduced in mule deer (Odocoileus hemionus) and white-tailed deer (O. virginianus) consuming tanniferous forages, digestion of the plant cell wall is not reduced beyond that predicted from its content of lignin, cutin, and silica. The lack of a tannin effect on cell wall digestion in deer is in contrast to studies with domestic sheep and numerous in vitro studies. Herbivores adapted to consume tanniferous forages may defend against such allelochemicals by producing salivary proteins that bind tannins in a highly specific manner. These tannin-salivary protein complexes would reduce apparent digestibilities of protein and cell solubles and, if completely effective, would not reduce cell wall digestion. The occurrence of such proteins in ruminants is reported here for the first time. The saliva composition of mule deer (a mixed feeder that commonly consumes browse) and domestic cattle and sheep (predominant grazers) are compared, and the higher potential of the deer saliva to neutralize tannins is related to their feeding habits. Salivary proteins that preferentially bind tannins may minimize fecal nitrogen losses by maximizing the efficiency of tannin-binding per unit of protein and may reduce the absorption of hydrolyzable tannins and the potential for tannin toxicity.

Epitope mapping.

This article describes a strategy for the mapping of the binding site, or epitope, of a monoclonal antibody (MAb) using bacterially expressed protein products. An overall strategy is discussed. This includes an initial round of several parallel approaches to gain the greatest amount of information at this stage. The second round uses the mapping information generated to identify MAbs, which may bind to identical or overlapping epitopes. The third round involves the design of new constructs that express small defined regions of the protein to refine the position of the epitope. The final step leads to the identification of the epitope to a resolution of 10 amino acid residues or else.

Full-field ERG in patients with Batten/Spielmeyer-Vogt disease caused by mutations in the CLN3 gene.

PURPOSE: To investigate, using full-field ERG, the retinal function in patients with Batten/Spielmeyer-Vogt disease caused by mutations in the CLN(3) gene. METHODS: Batten disease status of five patients was confirmed by the presence of vacuolated lymphocytes in peripheral blood and the identification of mutations in the Batten disease gene (CLN(3)). Visual acuity, fundus appearance, and full-field ERG were examined in all patients (age 4-19 years). The examination was repeated in one patient after 16 months. RESULTS: Three unrelated patients were homozygous for the most common mutation in CLN(3), the 1.02 kb deletion; two patients (sisters) were heterozygous for the 1.02 kb deletion and an as yet unidentified mutation in the CLN(3) gene. Full-field ERG recordings in all five patients demonstrated no rod responses and only small remaining cone responses, which could be detected with 30 Hz-flicker stimulation. Re-examination of a six-year-old girl after 16 months revealed a fast progression of the retinal degeneration. CONCLUSION: Full-field ERG recordings in Batten disease patients, both homozygous and heterozygous for the 1.02 kb deletion in the CLN( 3) gene, confirm retinal degeneration to be severe, widespread, and with a rapid progression early in the disease course. The onset of visual failure may be delayed when compared to the classic disease course, particularly in patients who are not homozygous for the most common CLN(3) mutation, a 1.02 kb deletion. In that case, the disease progression in terms of other symptoms may also be further delayed.

Epitope mapping.

Monoclonal antibodies (MAbs) are specific immunological tools because they bind to a precise determinant (the epitope) on the surface of a protein. The procedure of identifying the binding site of a MAb is often termed "epitope mapping."

Epitope mapping.

Monoclonal antibodies (MAbs) are specific immunological tools because they bind to a precise determinant (the epitope) on the surface of a protein. The procedure of identifying the binding site of a MAb is often termed "epitope mapping."

Genomic structure of three CLN3-like genes in Caenorhabditis elegans.

The genome of Caenorhabditis elegans is predicted to carry three genes similar to CLN3, the gene underlying juvenile neuronal ceroid lipofuscinosis. All three genes are transcribed and the genomic structure has been determined. The number and position of exons for two of the genes differ from that predicted from the genomic sequence, but no discrepancies with the genomic nucleotide sequence were found. Gene F07B10.1 (cln-3.1) is predicted to have 7 exons and to encode a protein of 424 amino acids. Gene C01G8.2 (cln-3.2) has 9 exons and encodes a protein of 435 amino acids. Gene ZC190.1 (cln-3.3) is predicted to have 9 exons and to encode a protein of 416 amino acids.

Analysis of CLN3-protein interactions using the yeast two-hybrid system.

Juvenile neuronal ceroid lipofuscinosis (Batten disease) is a childhood neurodegenerative disease that is caused by mutations in the CLN3 gene. The protein encoded by CLN3 has no homology with any proteins of known function and its cellular role remains elusive. In order to investigate the role played by the CLN3 protein we aimed to identify interacting proteins. Here, we describe the yeast two-hybrid system as the approach taken to investigate such protein-protein interactions. CLN3 was expressed as a fusion protein with a DNA-binding domain and used to screen a library of human fetal brain cDNAs fused to a transcriptional activation domain. Owing to low level expression of the full length CLN3 fusion protein, truncated regions corresponding to the predicted hydrophilic regions were also tested. No proteins that interact with CLN3 were detected, nor was there any evidence for CLN3-CLN3 interactions. Potential interaction of CLN3 with subunit c of mitochondrial ATP synthase, the major component of the storage material that accumulates in Batten disease patients, was also tested. No interaction was detected suggesting that the accumulation of subunit c does not result from loss of a process that requires a direct interaction with CLN3. We conclude that either CLN3 does not interact with other proteins or such interactions cannot be detected using the two-hybrid system.

Analysis of candidate genes in the CLN6 critical region using in silico cloning.

CLN6, the gene for variant late infantile neuronal ceroid lipofuscinosis, was mapped to a 4 cM region on chromosome 15q22-23. Subsequently the critical region was narrowed to less than 1 cM between microsatellite markers D15S988 and D15S1000 by additional marker typing in an expanded family resource. A physical map was constructed across this region using YAC and PAC clones and sequence was generated from two PAC clones. This sequence was analysed together with overlapping sequence generated by the Human Genome Project to identify genes within the region using an in silico cloning approach. In all, 29 genes have been identified and 18 have been analysed for mutations by direct sequencing. This powerful new approach will lead to the identification of CLN6.

Turkish variant late infantile neuronal ceroid lipofuscinosis (CLN7) may be allelic to CLN8.

One variant form of late infantile neuronal ceroid lipofuscinosis (LINCL) is found predominantly within the Turkish population (CLN7). Exclusion mapping showed that CLN7 was not an allelic variant of known NCL loci (CLN1, CLN2, CLN3, CLN5 or CLN6). Using the method of homozygosity mapping, a genome-wide search was undertaken and a total of 358 microsatellite markers were typed at an average distance of about 10 cM. A region of shared homozygosity was identified on chromosome 8p23. This telomeric region contained the recently identified CLN8 gene. A missense mutation in CLN8 causes progressive epilepsy with mental retardation (EPMR) or Northern epilepsy, which has so far been reported only from Finland and is now classified as an NCL. The mouse model mnd has been shown to carry a 1 bp insertion in the orthologous Cln8 gene. Statistically significant evidence for linkage was obtained in this region, with LOD scores > 3, assuming either homogeneity or heterogeneity. Flanking recombinants defined a critical region of 14 cM between D8S504 and D8S1458 which encompasses CLN8. This suggests that Turkish variant LINCL, despite having an earlier onset and more severe phenotype, may be an allelic variant of Northern epilepsy. However mutation analysis has not so far identified a disease causing mutation within the coding or non-coding exons of CLN8 in the families. The Turkish variant LINCL disease-causing mutation remains to be delineated.

New mutations in the neuronal ceroid lipofuscinosis genes.

Thirty-eight mutations and seven polymorphisms have recently been reported in the genes underlying the neuronal ceroid lipofuscinoses (NCLs) including 11 new mutations described here. A total of 114 mutations and 28 polymorphisms have now been described in the five human genes identified which cause NCL. Thirty-eight mutations are recorded for CLN1/PPT; 40 for CLN2/TTP-1, 31 for CLN3, four for CLN5, one for CLN8. Two mutations have been described in animal genes (cln8/mnd, CTSD). All mutations in NCL genes are contained in the NCL Mutation Database (http://www.ucl.ac.uk/NCL).

A critical analysis of techniques for measuring tannins in ecological studies : I. Techniques for chemically defining tannins.

A series of seventeen plant extracts rich in phenolic materials, including condensed and hydrolysable tannins, have been subjected to a series of chemical analyses in an attempt to gather ecologically significant information about their structure. Procedures investigated were (i) the Folin-Denis and Hagerman and Butler methods for quantifying total phenolics, (ii) the vanillin and proanthocyanidin methods for quantifying condensed tannins, (iii) the iodate and nitrous acid methods for hydrolysable tannins. It was found that the techniques for total phenolics correlated well, the Hagerman and Butler method giving higher estimates where solutions were particularly phenol rich. By contrast there was considerable discrepancy between the methods examined for condensed tannins. This is probably due primarily to the very different chemical reactions that form the basis of these procedures and also to the fact that the extract dependent products of the proanthocyanidin method vary in their A 11 values. During the study of condensed tannins methods for estimating their contribution to total phenolics and for measuring their average polymer length were examined. In both cases different procedures produced very variable results. Available methods for hydrolysable tannins were found not to be generally applicable across all extracts thought to contain this type of tannin on the basis of chromatographic analysis. An attempt to produce a quantitative spectrophotometric assay for hydrolysable tannins based on changes in reactivity to ferric chloride due to hydrolysis is described. This proved to be of limited sensitivity but may have some merit for estimating levels in hydrolysable tannins in phenol-rich plant extracts that also contain condensed tannins. It is concluded that whilst the overall level of phenolics in extracts can be estimated with some confidence the information imparted by more specific assays is very dependent on the procedures employed, particularly when dealing with extracts from taxonomically highly diverse sources.

A critical analysis of techniques for measuring tannins in ecological studies : II. Techniques for biochemically defining tannins.

A series of seventeen taxonomically diverse plant extracts rich in phenolic materials, including condensed and hydrolysable tannins, have been subjected to a series of biochemical analyses in an attempt to gather ecologically significant information about their interaction with proteins and amino acids. Methods employed were (i) protein-precipitation, using bovine serum albumin as substrate, followed by computation of specific activities of the tannins present in the extracts, and (ii) the inhibition of cellulase activity by tannin extracts bound to the cellulose substrate and free in solution. Both techniques revealed that all extracts contained tannin material. However, attempts to relate the results of the two procedures and in turn to relate them to information reported previously on the chemical properties of these extracts revealed that there was little correlation between any of the chemical or biochemical properties examined. From this analysis it would seem that whilst the analytical procedures available for studying tannins may generate ecologically useful information it is at present impossible, at least where plant material that is taxonomically diverse is being examined, to extrapolate from one type of measure to anticipate what would be observed from another type of measure. In addition to the above three other observations arose from this study. First, it appears to be generally true that there is not an absolute positive correlation between the level of protein precipitation and the incorporation of tannin in the tannin-protein precipitate. As relative protein concentration increases the proportion of tannin bound in the precipitate decreases, leading to less stable precipitates. Second, it is confirmed that some basic amino acids will precipitate with tannins, a phenomenon that could potentially influence amino acid balance in the diet. Third, complexation between tannin and protein absorbed on a cellulose substrate is able to interfere with the digestion of that cellulose by cellulase enzymes. Cellulose masking of this type may potentially effect the efficiency of cellulolytic activity in the rumen and if so suggests another subtle variation in the potential antifeedant properties of tannins.