French validation of the Foot Function Index (FFI).

OBJECTIVE: French validation of the Foot Function Index (FFI), self-questionnaire designed to evaluate rheumatoid foot according to 3 domains: pain, disability and activity restriction. METHODS: The first step consisted of translation/back translation and cultural adaptation according to the validated methodology. The second stage was a prospective validation on 53 patients with rheumatoid arthritis who filled out the FFI. The following data were collected: pain (Visual Analog Scale), disability (Health Assessment Questionnaire) and activity restrictions (McMaster Toronto Arthritis questionnaire). A test/retest procedure was performed 15 days later. The statistical analyses focused on acceptability, internal consistency (Cronbach's alpha and Principal Component Analysis), test-retest reproducibility (concordance coefficients), external validity (correlation coefficients) and responsiveness to change. RESULTS: The FFI-F is a culturally acceptable version for French patients with rheumatoid arthritis. The Cronbach's alpha ranged from 0.85 to 0.97. Reproducibility was correct (correlation coefficients>0.56). External validity and responsiveness to change were good. CONCLUSION: The use of a rigorous methodology allowed the validation of the FFI in the French language (FFI-F). This tool can be used in routine practice and clinical research for evaluating the rheumatoid foot. The FFI-F could be used in other pathologies with foot-related functional impairments.

Patient education after amputation: systematic review and experts' opinions.

OBJECTIVE: To perform a systematic review of the literature regarding amputee self-care, and analyze current experts' opinions. METHOD: The research in Medline and Cochrane Library databases was performed using the keywords "amputee self-care", "amputee health care", "amputee education", and "amputee health management". The methodological quality of the articles was assessed using four levels of evidence and three guideline grades (A: strong; B: moderate; C: poor). RESULT: One prospective randomized controlled study confirm the level of evidence of self-care amputee persons with grade B, which is similar others chronic diseases self-care. Self-care of amputee persons contributes to improve functional status, depressive syndrome, and also health-related quality of life. A review of the patients' needs and expectations in self-care amputee persons has been established thanks to the presence of qualitative focus group study. CONCLUSION: A multidisciplinary self-care of amputee persons can be recommended. Regarding literature date, the level of evidence of self-care amputee persons is moderate (grade B). Experts groups are currently working on a self-care amputee persons guideline book in order to standardize practicing and programs in the physical medicine and rehabilitation departments.

Isolation of a cDNA encoding a beta-1,4-endoglucanase in the root-knot nematode Meloidogyne incognita and expression analysis during plant parasitism.

A beta-1,4-endoglucanase encoding cDNA (EGases, E.C. 3.2.1.4), named Mi-eng-1, was cloned from Meloidogyne incognita second-stage juveniles (J2). The deduced amino acid sequence contains a catalytic domain and a cellulose-binding domain separated by a linker. In M. incognita, the gene is transcribed in the migratory J2, in males, and in the sedentary adult females. In pre-parasitic J2, endoglucanase transcripts are located in the cytoplasm of the subventral esophageal glands. The presence of beta-1,4-endoglucanase transcripts in adult females could be related to the expression of the gene in esophageal glands at this stage. However, cellulase activity within the egg matrix of adult females suggests that the endoglucanase may also be synthesized in the rectal glands and involved in the extrusion of the eggs onto the root surface. The maximum identity of the predicted MI-ENG-1 catalytic domain with the recently cloned cyst nematode beta-1,4-endoglucanases is 52.5%. In contrast to cyst nematodes, M. incognita pre-parasitic J2 were not found to express a beta-1,4-endoglucanase devoid of a cellulose-binding domain.

Molecular cloning of an acetylcholinesterase gene from the plant parasitic nematodes, Meloidogyne incognita and Meloidogyne javanica.

A gene encoding a protein with strong homology with Caenorhabditis elegans and C. briggsae acetylcholinesterase ACE-1 was cloned from Meloidogyne incognita and M. javanica pre-parasitic juveniles. Both cDNAs have an ORF of 1968 bp for a deduced translation product of 656 amino acid residues. The key residues essential to acetylcholinesterase (AChE) structure and function are conserved in both sequences. M. incognita and M. javanica AChE share a homology of 98.8% at the amino acid level and 97% at the nucleotide level. Phylogenetic analysis showed that Meloidogyne and Caenorhabditis AChE form a cluster among AChE of triploblastic organisms. This Meloidogyne AChE is expressed in eggs, pre-parasitic juveniles and males and AChE activity was detected in situ in amphids of pre-parasitic juveniles. The opportunity of using AChE as a target in new strategies of nematode control is discussed.

Identification, characterisation and cDNA cloning of two caseins from the common brushtail possum (Trichosurus vulpecula)1.

Two major caseins have been isolated from the milk of the common brushtailed possum (Trichosurus vulpecula). These have been identified as alpha- and beta-casein on the basis of the similarity of their N-terminal sequences to those of the caseins of another marsupial (Macropus eugenii). Both proteins appear to exist in multiple forms. Possum alpha-casein is glycosylated mainly in the form of sialic acid residues and was shown by electrospray mass spectrometry to have multiply phosphorylated forms of three families with molecular masses 22700 and 23200 Da that may represent genetic variants. Two-dimensional electrophoresis showed that beta-casein exists as a complex of five or six proteins of identical N-terminal sequence but differing pI. Electrospray mass spectrometry indicated that the beta-caseins also are multiply phosphorylated with masses between 32300 and 32600 Da. A subfamily with mass values 1530 greater was also detected. The patterns were not affected by stage of lactation and quantitative analysis of two-dimensional gels of whole milk shows that alpha- and beta-caseins are present at a constant ratio throughout lactation. cDNA clones for the possum alpha- and beta-caseins have been isolated from an early lactation mammary cDNA library and sequenced.

Phylogenetic analysis of three lipocalin-like proteins present in the milk of Trichosurus vulpecula (Phalangeridae, Marsupialia).

Three proteins have been identified in the milk of the common brush tail possum. Trichosurus vulpecula that from sequence analysis are members of the lipocalin family. They include beta-lactoglobulin, which appears to have two forms; a homologue to the late-lactation protein found in tammar, Macropus eugenii; milk; and a novel protein termed trichosurin. Whereas beta-lactoglobulin and trichosurin are both expressed throughout lactation, the late-lactation protein is not detected in samples taken before days 100-110 of lactation. The cDNAs encoding each of these proteins have been isolated from cDNA libraries prepared using possum mammary mRNA and sequenced. Phylogenetic analysis showed that the T. vulpecula beta-lactoglobulin, along with two other macropod beta-lactoglobulins, forms a subclass of beta-lactoglobulins distinct from those for eutherian mammals; both marsupial late-lactation proteins appear to have similarities to a family of odorant-binding proteins, whereas trichosurin has similarities to the major urinary proteins of rodents.

Differential expression of milk protein genes during lactation in the common brushtail possum (Trichosurus vulpecula).

In the common brushtail possum (Trichosurus vulpecula) lactation lasts for 200 days and consists of two distinct phases. Milk composition changes dramatically between phase 2 and 3, which correspond to early and late lactation respectively (phase 1 corresponds to pregnancy). RNA expression patterns have been established for eight major milk protein genes throughout lactation in possum mammary glands. The levels of mRNA expressed from two genes, encoding the early and late lactation proteins, were differentially regulated during lactation, with peak RNA levels occurring in phase 2 and 3 of lactation respectively. Expression of these two RNA transcripts did not overlap, and neither gene was expressed at significant levels between days 116 to 125, suggesting that the transition from phase 2 to phase 3 of lactation occurs at this time. The level of lysozyme, alpha-lactalbumin and trichosurin mRNA increased in phase 3 of lactation, whereas the levels of beta-lactoglobulin, alpha-casein and beta-casein mRNA remained constant throughout lactation. In the non-suckled gland, expression of milk protein genes was greatly reduced by day 6 of lactation. In conclusion, the early and late lactation protein genes are good markers for phase 2 and 3 of lactation, with the transition between these phases occurring around day 120 of lactation in the possum.

Lysozyme and alpha-lactalbumin from the milk of a marsupial, the common brush-tailed possum (Trichosurus vulpecula).

Lysozyme and alpha-lactalbumin have been identified using N-terminal sequence analysis of whey proteins from the common brush-tailed possum, Trichosurus vulpecula after separation by two-dimensional denaturing electrophoresis. Both proteins were purified from pooled possum milk using ion exchange chromatography and gave mass values of 14,896 and 13,985 Da respectively by MALDI-TOF mass spectrometry. Clones containing the full coding sequences of the genes for both proteins were isolated from a possum mammary cDNA library and the DNA sequence of the coding region determined. The inferred protein sequences were used in phylogenetic analysis of both protein classes. These showed that the T. vulpecula alpha-lactalbumin, along with other marsupial alpha-lactalbumins, formed a family distinct from the eutherian alpha-lactalbumins and the alpha-lactalbumin of a monotreme, the platypus, consistent with the separate evolution of the marsupials. By contrast the T. vulpecula lysozyme was shown to be similar to the ruminant stomach lysozymes and primate lysozymes and quite distinct from the Ca2+-binding lysozymes found in the milk of the echidna and horse.

A novel marsupial protein expressed by the mammary gland only during the early lactation and related to the Kunitz proteinase inhibitors.

A novel whey protein has been found in marsupial milk, the early lactation protein (ELP). The whey of Trichosurus vulpecula, the Australian common brush-tailed possum, contains two forms of ELP, estimated by protein electrophoresis at 8 and 16 kDa. The 16-kDa form contains approximately 60% N- linked carbohydrate. The ELP cDNA was obtained by the screening of an early lactation cDNA library with an early lactation total cDNA probe and random selection of strongly positive clones. The full-length cDNA sequence of 306 bp codes for an 82 amino acid residue mature protein and a 20-residue secretory signal peptide. The mature protein has a calculated molecular weight of 9325.4 and a pI of 8.1. Protein and RNA analysis show that the expression of the ELP is restricted only to the early lactation phase. The ELP has amino acid sequence homologies with the Kunitz proteinase inhibitor family and the whey acidic proteins.

Cloning and characterization of two satellite DNAs in the low-C-value genome of the nematode Meloidogyne spp.

Two highly reiterated StyI satellite DNAs have been cloned from two nematode species: one from Meloidogyne hapla and another from M. incognita. The monomeric units of these two satellites have a repeat length of 169 and 295 bp, respectively. These StyI repeated element families constitute 5% of the M. hapla and 2.5% of the M. incognita haploid genomes. The A + T content is elevated in both families (i.e., 68% and 77%, respectively). Nucleotide methylation and transcriptional activity are negative. No similarity was found between the two satellites, nor to other known highly repetitive elements. These StyI satellite DNAs are quite homogenous in sequence, showing on average 3% and 3.5% divergence from their respective calculated consensus sequence. An internal subrepeating unit of about 11 bp is observed in the StyI satellite monomer sequences of M. hapla, suggesting that it could have evolved from a shorter sequence. Because of the small size of the Meloidogyne genome (51 Mb) and the abundance of repeated sequences, this genus approaches a limit in terms of coding fraction.

Sequences homologous to Tc(s) transposable elements of Caenorhabditis elegans are widely distributed in the phylum nematoda.

To have a better understanding of the evolutionary history of mobile elements within the nematodes, we examined the distribution and the conservation of homologues to transposable elements from Caenorhabditis elegans (Tc1, Tc2, Tc3, Tc4, Tc5, and FB1) in 19 nematode species belonging to the class Secernentea. Our results show that Tc1 elements display a distribution restricted to the family Rhabditidae with poor conservation. The Tc2 and FB1 homologous elements have the same patchy distribution within the Rhabditidae. They were only found in Caenorhabditis and in Teratorhabditis. The Tc3 element is widely distributed among nematode species. Tc3 homologous elements are present in the majority of the Rhabditidae but also in two genera within the family Panagrolaimidae, and in Bursaphelenchus, which belongs to the order Aphelenchida. Tc4 and Tc5 homologues show the most limited distribution of all tested elements, being strictly limited to C. elegans. These data indicate that in some cases, the distribution of transposable elements in the nematode cannot be explained by strict vertical transmission. The distribution of Tc3, Tc4, and Tc5 suggests that horizontal transmission may have occurred between reproductively isolated species during their evolutionary history.

Isolation of a Repeated DNA Probe Showing Polymorphism among Meloidogyne incognita Populations.

Several Meloidogyne incognita geographic populations were characterized by analysis of the restriction fragment length polymorphisms (RFLP) obtained after digestion of their total DNA and hybridization with a [(3)(2)P]-labeled probe. The probe consisted of a 1.7-kb-repeated DNA sequence, isolated from a M. incognita genomic library, that hybridized to multiple BamH I fragments in the genome of each isolate. The patterns showed sufficient polymorphism to enable the accurate differentiation of all the populations tested.