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[Injuries of the dens axis in childhood. Biomechanical analysis and surgical and conservative treatment of 2 cases].

Odontoid "fractures" in children are in fact typical lesions of the cartilaginous plate ("synchondrosis") separating the odontoid process from the body of the axis, 54 cases of which have been described in the literature so far. In our review we report about two 2-year-old children who were back-seat passengers restrained by four-point children's seat belts, in a car involved in a head-on motor vehicle accident. According to the accident research unit of our institution, trauma sustained in such circumstances is adequate for the causation of odontoid lesions in healthy children. Head-on collisions at a speed absorption of at least 40 km/h is described as a typical mechanism of such injury. Both children were immediately symptomatic, and the diagnosis was easily made from the X-rays. Neither child had neurologic deficits, which is in keeping with the literature, where neurologic injuries were found to have been reported exclusively in conjunction with head injuries. After closed reduction, conservative treatment was initiated in one child, in whom the extent of the anterior dislocation was smaller than the diameter of the odontoid shaft. In cases of major dislocation and more pronounced instability we recommend primarily open reduction and osteosynthesis with adequate implants. Our second case was treated with a halo fixator and a plaster vest for 12 weeks, but despite anatomic reduction there was no healing of the odontoid process. After temporary posterior fusion of C1/C2 we reamed the synchondrosis from an anterior approach and performed autogenous bone grafting. The posterior cerclage wire was removed after 5 months.(ABSTRACT TRUNCATED AT 250 WORDS)

Accidents, Traffic↗

[Analysis of recurrences following conservative treatment of breast carcinoma].

In the conservative treatment of early breast cancer, great attention must be paid to define the prognostic factors correlated with the local recurrence rate. The goal is to customize surgery, radiotherapy and chemotherapy to the risk predicted in every single patient. To investigate the impact of some prognostic factors in a group of patients treated with homogeneous treatment schedules, 251 women with UICC stage I or II breast cancer were examined in the Padua Radiotherapy Department from 1988 to 1990. All patients underwent conservative surgery consisting in quadrantectomy, axillary node dissection and radiotherapy. During a median follow-up period of 49.2 months, 12 patients presented a breast relapse (4.8%). In 4 patients the relapse occurred in the same quandrant as the primary lesion, whereas a different quadrant was involved in the other 8 patients. The relapse rate in women under 60 was 5% and 4.7% in older patients, with p = 0.73. In pT1 patients, the relapse rate was 4.5% and in pT2 patients it was 7.9% (p = 0.37). No significant difference was observed between pN- and pN+ patients (4.5% vs. 6.25%, p = 0.37). In our series, none of the studied factors significantly influenced breast relapse rates. The number of patients may be too little relative to the low rate of relapses. However, an unfavorable trend was observed in the patients under 60, in pT2 or pN+ patients, or in the patients with positive or unknown surgical margins.

Adult↗

Molecular evolution of PAS domain-containing proteins of filamentous cyanobacteria through domain shuffling and domain duplication.

When the entire genome of a filamentous heterocyst-forming N2-fixing cyanobacterium, Anabaena sp. PCC 7120 (Anabaena) was determined in 2001, a large number of PAS domains were detected in signal-transducing proteins. The draft genome sequence is also available for the cyanobacterium, Nostoc punctiforme strain ATCC 29133 (Nostoc), that is closely related to Anabaena. In this study, we extracted all PAS domains from the Nostoc genome sequence and analyzed them together with those of Anabaena. Clustering analysis of all the PAS domains gave many specific pairings, indicative of evolutionary conservations. Ortholog analysis of PAS-containing proteins showed composite multidomain architecture in some cases of conserved domains and domains of disagreement between the two species. Further inspection of the domains of disagreement allowed us to trace them back in evolution. Thus, multidomain proteins could have been generated by duplication or shuffling in these cyanobacteria. The conserved PAS domains in the orthologous proteins were analyzed by structural fitting to the known PAS domains. We detected several subclasses with unique sequence features, which will be the target of experimental analysis.

Amino Acid Sequence↗

Phylogeny of capsid proteins of rod-shaped and filamentous RNA plant viruses: two families with distinct patterns of sequence and probably structure conservation.

Computer-assisted comparative analysis of all available amino acid sequences of the capsid proteins of positive strand RNA plant viruses with helical capsids is described. Two distinct families of homologous proteins were delineated through statistically significant sequence similarities, one including the capsid proteins of rod-shaped viruses (tobamo-, tobra-, hordei-, and furoviruses) and the other those of filamentous viruses (poty-, bymo-, potex-, carla-, and closteroviruses). It was concluded that the capsid proteins of all rod-shaped viruses, on the one hand, and filamentous viruses, on the other hand, evolved from common ancestors. Analysis of residue conservation patterns in the capsid proteins of rod-shaped viruses revealed maintenance of the hydrophobic core and of the (putative) salt bridge between conserved Arg and Asp residues. Sequence comparisons within the filamentous virus family expanded the observations on the relationship between the capsid proteins of potex-, carla-, poty-, and bymoviruses. Grouping of the beet yellows closterovirus capsid protein sequence, recently determined in this laboratory (Agranovsky et al., J. Gen. Virol., 1991, 72, 15-23), with those of potex- and carlaviruses was demonstrated. The coat protein of another closterovirus, apple chlorotic leaf spot virus, appeared to constitute a distinct phylogenetic lineage. Despite the lack of significant overall similarity, comparison of the alignments of the capsid proteins of the two families suggested formation of analogous salt bridges.

Amino Acid Sequence↗

High affinity YY1 binding motifs: identification of two core types (ACAT and CCAT) and distribution of potential binding sites within the human beta globin cluster.

PCR-assisted binding site selection was used to define the sequence characteristics of high affinity YY1 binding sites. Compilation of the sequences of 189 selected oligonucleotides containing high affinity YY1 binding sites revealed two types of core sequence: ACAT and CCAT. ACAT cores were surrounded by other invariant nucleotides, forming the consensus GACATNTT. A search of the 73 kb human beta-like globin cluster with this consensus revealed eight matching motifs, six of which were located within 1-3 kb upstream of the gamma and beta genes. CCAT-type cores were more variable in surrounding sequence context; the consensus VDCCATNWY was found to fit 89% of the selected CCAT-containing oligonucleotides. A search of the human beta globin cluster with CCAT consensus sequences revealed 171 potential YY1 binding sites. Several of these were tested directly in gel shift assays and confirmed as high affinity YY1 binding sites. Finally, a strategy called motif-based phylogenetic analysis was employed to determine which of the 179 total sites are evolutionarily conserved. This analysis permits the detection of functionally conserved binding sites despite sequence differences present between the two species. The 21 conserved sites identified will serve as important starting points in further dissection of the possible role of YY1 in globin gene regulation.

Animals↗

Isolation of two cDNAs encoding novel alpha 1-antichymotrypsin-like proteins in a murine chondrocytic cell line.

We have isolated two novel serpin-encoding sequences from EB22, a chondrocytic cell line derived from a mouse teratocarcinoma. Both sequences fall within the Spi-2 sub-family, and are related to the gene encoding human alpha 1-antichymotrypsin (ACT), a major acute-phase reactant. Considerable amplification of the Spi-2 gene family in the mouse has occurred, hindering the identification of a functional equivalent of the human gene. However, one of the sequences described here, EB22/4, exhibits several features which indicate that it may represent the physiological rodent equivalent of ACT. The sequence is expressed in the liver, as expected, and is induced several-fold during the acute-phase response. The P1 amino acid residue, which is primarily responsible for inhibitor specificity, is Met rather than the human Leu, most probably a functionally conservative substitution. Analysis of the orthologous sequence in related rodents demonstrates conservation of the predicted reactive centre-encoded specificity. The second isolated cDNA, EB22/3, encodes an unexpected Cys residue at the P1 position in the reactive centre, and represents a novel sub-class of the Spi-2 serine proteinase inhibitor (serpin)-encoding gene family. At least one of the sequences appears to be expressed at sites of skeletal deposition during the later stages of mouse foetal development, indicating a role for serpins during development.

Acute-Phase Proteins↗

What does an intermediate success rate mean? An analysis of a Piagetian liquid conservation task in the great apes.

The study investigates what an intermediate success rate means in bonobos, chimpanzees, and orangutans. Apes participated in liquid conservation experiments where they had to track the larger of two different quantities of juice after various kinds of transformations [Suda, C., & Call, J. (2004). Piagetian liquid conservation in the great apes (Pan paniscus, Pan troglodytes, and Pongo pygmaeus). Journal of Comparative Psychology, 118, 265-279). When making a decision, apes sometimes demonstrated hesitant behavior, concurrently pointing to both alternatives or successively changing their choice. Moderately successful apes showed more hesitation than highly successful or unsuccessful apes. The results are consistent with the cognitive conflict model: The experiments created a higher degree of cognitive conflict on moderately successful apes than on very successful or unsuccessful apes. This indicates that an intermediate performance reflects the joint operation and potential conflict between two different cognitive strategies (identity and appearance) inherent to the Piagetian conservation task.

Analysis of Variance↗

A new domain family in the superfamily of alkaline phosphatases.

During the course of our large-scale genome analysis a conserved domain, currently detectable only in the genomes of Drosophila melanogaster, Caenorhabditis elegans and Anopheles gambiae, has been identified. The function of this domain is currently unknown and no function annotation is provided for this domain in the publicly available genomic, protein family and sequence databases. The search for the homologues of this domain in the non-redundant sequence database using PSI-BLAST, resulted in identification of distant relationship between this family and the alkaline phosphatase-like superfamily, which includes families of aryl sulfatase, N-acetylgalactosomine-4-sulfatase, alkaline phosphatase and 2,3-bisphosphoglycerate-independent phosphoglycerate mutase (iPGM). The fold recognition procedures showed that this new domain could adopt a similar 3-D fold as for this superfamily. Most of the phosphatases and sulfatases of this superfamily are characterized by functional residues Ser and Cys respectively in the topologically equivalent positions. This functionally important site aligns with Ser/Thr in the members of the new family. Additionally, set of residues responsible for a metal binding site in phosphatases and sulphtases are conserved in the new family. The in-depth analysis suggests that the new family could possess phosphatase activity.

Alkaline Phosphatase↗

Conservative management of acoustic neuroma: a meta-analysis and proposed treatment algorithm.

OBJECTIVES/HYPOTHESES: Conservative management is a viable treatment alternative for acoustic neuroma. Using previous studies to provide evidence-based support, we have attempted to more clearly define the role of conservative management. STUDY DESIGN: Retrospective review of literature and patient charts. METHODS: Published studies on conservative management of acoustic neuroma were found using a key word search through PubMed in addition to the bibliographies of these selected studies. A spreadsheet was made to tabulate the selection criteria for conservative management, duration and frequency of follow-up, patient demographics, initial tumor size and rate of growth, change in hearing status, and the need for definitive treatment. RESULTS: A total of 21 studies comprising 1,345 patients were included in our meta-analysis. The average length of follow-up these studies was 3.2 years. The average initial tumor size was 11.8 mm (n = 900); 43% of 1,244 acoustic neuromas showed growth, whereas 57% showed either no growth or tumor regression. The average growth rate was 1.9 mm/year in 793 individuals. Hearing loss occurred in 51% of 347 individuals. In 15 studies, 20.0% of 1,001 individuals eventually failed conservative management. CONCLUSIONS: Our meta-analysis supports the role of conservative management of acoustic neuromas in properly selected patients on the basis of a slow overall rate of growth and a substantial incidence of no growth. However, the lack of predictive factors, the relatively short duration of follow-up, and the variability of inclusion criteria underscore the need for continued collection of long-term data. An algorithm for acoustic neuroma management is proposed based on initial tumor size, patient age, and hearing status.

Algorithms↗

Tandem binding of six OmpR proteins to the ompF upstream regulatory sequence of Escherichia coli.

OmpR is a transcription factor in Escherichia coli whose function is modulated by phosphorylation in the presence of phosphorylated EnvZ, a transmembrane protein histidine kinase involved in osmosensing. Using a protein S-OmpR hybrid protein, we demonstrated that six OmpR molecules bind tandemly to the -100 to -39 sequence of ompF. This sequence consists of three 20-base pair units: F1, F2, and F3, each of which is bound by two OmpR proteins. Polymerase chain reaction selection of nine randomized base pairs within the F1 sequence revealed highly conserved C residues spaced 10 base pairs apart. Further mutational analysis of conserved bases indicated that two OmpR molecules bind tandemly to two direct repeats. Mobility shift assays showed that cooperative interactions play a role in enhancing binding of OmpR to lower affinity F2 and F3 sites. Activation and repression of ompF expression are thus regulated by a total of eight OmpR molecules, including two molecules that bind to a distal site (-380 to -361).

Bacterial Outer Membrane Proteins↗

The mouse genome sequence--the end of the tail, or just the beginning?

The recent flurry of papers on the mouse genome includes the description of the full genome assembly, analysis of the mouse transcriptome, the origin of interstrain variation, initial analysis of conserved non-coding regions and high-throughput expression analysis of a subset of genes. Each illustrates how the availability of the genome sequence will change the way mouse biologists do business in future.

Alternative Splicing↗

A comparative analysis of translated dihydropyrimidine dehydrogenase cDNA; conservation of functional domains and relevance to genetic polymorphisms.

A pharmacogenetic syndrome caused by molecular defects in the dihydropyrimidine dehydrogenase gene (DPYD ) results in partial to complete loss of dihydropyrimidine dehydrogenase (DPD) enzyme activity with patients exhibiting life-threatening toxicity following administration of routine doses of 5-fluorouracil. To date, more than 19 reported mutations have been putatively associated with DPD deficiency with 16 occurring within the open reading frame of the cDNA. The purpose of this study was to examine the conservation of functional domains (including the uracil, flavine adenine dinucleotide and NADPH binding sites) across three phyla (Chordata, Arthropoda and Nematoda) and the conservation of regions corresponding to the previously reported mutations. Comparative analysis of the uracil and NADPH binding sites in mammals and invertebrates demonstrated 100% amino acid identity between mammals and Drosophila melanogaster. Caenorhabditis elegans demonstrated 89% and 88% identity in these domains, respectively. The mammalian sequences demonstrated 100% identity in two iron sulphur motifs (amino acids 953-964 and 986-997) with significant conservation in D. melanogaster (92% and 92% identity, respectively) and C. elegans (100% and 92% identity, respectively). Comparative amino acid analysis revealed non-conservation in the loci of four DPYD mutations [DPYD*12 (R21Q), DPYD*5 (I543V), DPYD*6 (V732I), DPYD*9A (C29R)]. Seven mutations occurred in highly conserved regions [M166V, DPYD*8 (R235W), DPYD*11 (V335l), DPYD*4 (S534N), DPYD*9B (R886H), D949V, DPYD*10 (V995F)]. In summary, this comparative analysis identified conserved regions which may be critical to enzyme structure and/or function. The conservation of loci where DPYD mutations occur was also examined to evaluate their functional significance on DPD enzyme activity. These data should prove useful in the evaluation of newly discovered mutations in the DPYD gene.

Amino Acid Sequence↗