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PubMed1999

[Digital film viewer versus conventional view boxes in the identification of bone and joint disorders].

PURPOSE: To compare the diagnostic accuracy of a digital film viewer (Smartlight 2000 Plus) versus a conventional view box for the identification of bone and joint disorders. MATERIAL AND METHODS: In order to evaluate the qualitative and quantitative differences of digital and conventional film viewers, 100 plain films of patients with bone and joint disorders taken in an emergency room January through May 1998 were reviewed utilizing both types of view boxes. Three radiologists expert of bone and joint disorders, independently compared the films, filled a form about the qualitative and quantitative analysis of the lesions depicted for each patient, and reported a few notes on the technical quality of the plain films in terms of exposure. The results were compared using the chi-square test (p < 0.005). RESULTS: Data analysis showed that the digital film viewer permitted a quicker reading of the film while decreasing the perception threshold for elementary lesions and ocular fatigue. All the radiographs read with the digital film viewer were considered technically adequate: the film quality was considered good in 10 cases and sufficient in 2. Two of the same radiographs read with the conventional view box were considered of good quality, nine were considered sufficient and one was considered insufficient due to overexposure. There were six cases of agreement and six of disagreement for the diagnosis: four were due to overexposure of the radiograph and two to better conspicuity provided by the digital film viewer. DISCUSSION: Correct image illumination is the first element a radiologist evaluates when reading a radiograph. A conventional view box may, when it is not properly maintained or when the radiograph is overexposed, decrease the radiologist's visual capacity. This increases the time required to read the radiograph and, therefore, physical and ocular fatigue, which in turn increases the risk of missing or misevaluating a lesion. A digital film viewer emits light with a variable intensity which is proportional to the optical density of the film. This increases the visual capacity of the radiologist and the lesion contrast, while reducing the factors which affect the radiograph reading. CONCLUSIONS: The use of a digital film viewer increased the visual capacity of the radiologist and eliminated the negative elements which complicated the radiograph reading and permitted the use of radiographs that would otherwise have been considered of poor quality due to overexposure. This, combined with the experience of the radiologist, decreased of the risk of missing or misevaluating a lesion.

Bone Diseases↗

PubMed2002

Efficient internalization of the polo-box of polo-like kinase 1 fused to an Antennapedia peptide results in inhibition of cancer cell proliferation.

Polo-like kinases (Plk) regulate multiple stages in mitosis. Plk1 is overexpressed in tumors. The COOH-terminal regions of Plks contain a conserved domain, termed polo-box, which is required for subcellular localization and for physical interaction with substrates. We linked the polo-box (amino acids 410-429) of Plk1 to an Antennapedia peptide and studied its impact on tumor cells. Whereas the wild-type polo-box inhibited the proliferation of tumor cells associated with induction of apoptosis, a mutated derivative was much less effective. The treatment caused mitotic arrest, misaligned chromosomes, and multiple centrosomes. Taken together, membrane-permeable polo-box peptides inhibit cancer cell proliferation efficiently.

Amino Acid Sequence↗

PubMed2002

Naf1 p is a box H/ACA snoRNP assembly factor.

Box H/ACA small nucleolar ribonucleoprotein particles (snoRNPs) contain four essential proteins, Cbf5p, Gar1p, Nhp2p, and Nop10p, each of which, with the exception of Gar1p, is required for box H/ACA snoRNA accumulation. Database searches identified a novel essential protein, which we termed Naf1p, with a region of homology to the RNA-binding domain of Gar1p and other features in common with hnRNP-like proteins. Naf1p is localized to the nucleus and is not a stable component of the H/ACA snoRNPs, but it is required for the accumulation of all box H/ACA snoRNAs. This requirement is not at the level of snoRNA transcription initiation or termination. Naf1 p shows in vitro RNA-binding activity and also binds directly to Cbf5p and Nhp2p. Naf1p was shown to bind to the CTD in vivo in a two-hybrid assay, and the phosphorylated CTD, but not the nonphosphorylated CTD, was shown to precipitate tagged Naf1p from a cell lysate. We propose that Naf1 p is recruited to the CTD of RNA polymerase II and binds to nascent box H/ACA snoRNAs promoting snoRNP assembly.

Amino Acid Motifs↗

PubMed1999

[Gene regulation studies on purine biosynthetic in Salmonella typhimurium IX. Mutation analysis of PUR box 2].

Two consensus bases C and G in 16 bp PUR box were directed mutated to G and A separately by PCR amplification. The binding function of PUR box carrying mutation with PUR protein were examined by gel retardation experiment. The results showed that the PUR box with above mutations could not bind with purR protein extracted from LT2. It proved that the two consensus bases C and G are necessary for PUR box binding with purR protein.

Bacterial Proteins↗

PubMed2002

Epidemiological typing of Streptococcus pneumoniae from various sources in India & Korea using Box A PCR fingerprinting.

BACKGROUND & OBJECTIVES: Epidemiological typing of Streptococcus pneumoniae is necessary to study genetic relatedness among strains. The Box A PCR assay, a good epidemiological tool for high resolution typing of pneumococcal isolates, was used for comparison of strains of S. pneumoniae from Korea, an area of high penicillin resistance and India with low resistance to penicillin. METHODS: The pneumococcal strains from Indian and Korean sources included in the study belonged to serotypes 19F, 23F, 6A & 6B. The penicillin susceptibility of the strains was confirmed by determination of minimum inhibitory concentration values. The strains were then fingerprinted using the Box A PCR protocol and the results were further analysed by Molecular Analyst Software. RESULTS: The Box fingerprinting technique produced sharp, clear and reproducible banding patterns for strains of S. pneumoniae included in the study. There was no similarity between Indian and Korean strains at the genetic level. Among the Korean strains with similar serotypes, the level of similarity varied. Except for one pair (serotype 23F), that showed a level of homology of about 90 per cent, almost all the others showed less than 80 per cent homology. It was also seen that there was a high percentage of homology (> 90% in nasopharyngeal isolates from India belonging to serotype 19F) between isolates from similar source. Comparison of the Indian and Korean isolates of similar serotype showed low homology, except for one cluster of two strains with serotype 6B which showed 84 per cent homology (blood isolates from India and Korea). INTERPRETATION & CONCLUSION: Box A PCR is a highly discriminatory and useful method for typing S. pneumoniae. The results of the study have shown low levels of homology between strains from within India and also between two geographically distinct areas.

Bacterial Typing Techniques↗

PubMed2004

[Cloning of a MADS box protein gene (GhMADS1) from cotton (Gossypium hirsutum L.)].

As a kind of transcription factors, MADS-box protein plays an important role in various cellular processes, especially in the development of floral organs. Based on the contig analysis of the cotton ESTs, the coding region of a cotton MADS-box protein (GhMADS1) was obtained by RT-PCR from floral buds of cotton (G. hirsutum). The cloned fragment of 713 bp (GhMADS1, GenBank accession no. AF538965) contains an open reading frame of 711 bp,coding a polypeptide of 236 amino acids. It was demonstrated that the deduced GhMADS1 protein was highly homologous to the AGL2 group of MADS-box proteins from Vitis vinifera, Nicotiana sylvestris, Petunia hybrida, Arabidopsis thaliana and Antirrhinum majus. Phylogenetic analysis also indicated that GhMADS1 belongs to the AGL2 group of MADS-box proteins. RT-PCR analysis showed that GhMADS1 gene expressed in petals, stamens, ovules and fibers, but not in roots, stems, leaves, bracts and sepals. The strongest expression of GhMADS1 gene was detected in petals. But in floral buds of a cotton homeotic mutant (CHV1), whose floral organs are all converted to bract leaf-like organs, the transcript of GhMADS1 gene was not detected. It was proposed that GhMADS1 gene would be crucial to the development of cotton floral organs.

Amino Acid Sequence↗

PubMed2005

[A method for Rhesus box test].

To study the method for Rhesus box test and its significance, the sequence specific primers of upstream, downstream and hybrid Rhesus boxes were designed according to RhD gene sequence; the upstream, downstream and hybrid Rhesus boxes were determined by PCP-SSP and mismatched PCR. The results showed that this method was confirmed by DNA Standard test. It was shown that in unrelative RhD positive individuals RHD(+)/RHD(-), RHD(+)/RHD(+) genotype accounted for 9.00%, 91.00% respectively, and in RhD negative individuals RHD(+)/RHD(-), RHD(+)/RHD(+), RHD(-)/RHD(-) genotype were 26.14%, 3.92%, 69.94% respectively. It is concluded that the method of Rhesus box test was confirmed to be reliable and can be used for the identification of RhD haplotype gene structure, as well as for study on inheritance, clinical transfusion and neonatal hemolytic diseases.

Base Sequence↗

PubMed1991

The MADS box gene family in tomato: temporal expression during floral development, conserved secondary structures and homology with homeotic genes from Antirrhinum and Arabidopsis.

Five genes with homology to the floral homeotic genes deficiens of Antirrhinum and agamous of Arabidopsis were isolated from tomato. Each of the five genes is unique in the genome and could be localized to a different chromosome by RFLP mapping. Four of the tomato genes (hereafter TM) are flower-specific with distinguishable temporal expression. TM4 and TM8 are 'early', while TM5 and TM6 are 'late' genes. TM4 is homologous to squamous and TM6 is similar to deficiens, which are, respectively, 'early' and 'late' bona fide homeotic genes in Antirrhinum. The proteins encoded by the five tomato genes, like several known homeotic genes from other plants, contain within their N-terminus a highly conserved DNA-binding domain, the MADS box. All known plant MADS box genes also share, however, other properties. They all contain a central, moderately conserved, and rather basic domain, and a highly divergent or even missing C-terminal domain. Furthermore, molecular modelling predicts the presence of a conserved amphipatic alpha helix, at a constant distance from the MADS box in each of these proteins. The common properties of eight MADS box proteins from three plant families indicate that all their domains were coded for by the same ancestor gene. The sequence homology between pairs of MADS genes from different species indicates that the MADS ancestor gene multiplied and diverged in an ancestor plant common to several dicotyledon families.

Amino Acid Sequence↗

PubMed1990

A cis-acting element located between the cAMP response elements and CCAAT box augments cell-specific expression of the glycoprotein hormone alpha subunit gene.

Several cis-acting elements interact through their cognate trans-acting factors to confer placenta-specific expression to the alpha subunit gene of glycoprotein hormones. These elements include two tandemly arranged cAMP response elements (CREs, located between base pairs (bp) -146 and -111 relative to the transcription start site), an upstream regulatory element (URE, located between -180 and -150), and a CCAAT box (located between bp -100 and -80). Here, we identify a new regulatory region, junctional regulatory element (JRE), with critical nucleotides located between the CREs and CCAAT box (bp -120 to -100). Although the binding site of the JRE abuts the 3' CRE, methylation interference assays indicate that no overlap occurs between the contact points of the proteins that bind to the JRE and CRE. This new regulatory element is highly conserved across species and contains a palindromic binding site for a 50-kDa nuclear factor(s) which is distinct from the factors that bind the URE, CRE, and CCAAT box of the alpha subunit gene. Finally, gene transfer studies suggest that, although JRE binding activity is found in several cell types, this element acts relatively cell-specifically. We conclude that this element and its cognate trans-acting factor act in conjunction with the complexes formed over the URE, CREs, and CCAAT box to enhance the placenta-specific activity of the alpha subunit promoter.

Base Sequence↗

PubMed1991

Defective HLA DRA X box binding in the class II transactive transcription factor mutant 6.1.6 and in cell lines from class II immunodeficient patients.

6.1.6 is one of several immunoselected mutants from EBV-transformed human B cell lines that have undergone coordinate loss of expression of all their HLA class II genes. Similar defects have been found in cells from some patients with class II immunodeficiencies. Previous studies have suggested that the defects in 6.1.6 and in the other class II regulatory mutants are in transactive factors required for class II transcription. The defective factors, however, have not been identified. Here we present two lines of evidence that serve to localize the site of action of the factor that is defective in 6.1.6. First, transfected indicator genes linked to HLA DRA promoter fragments that include the conserved X box region are transiently expressed at greatly reduced levels in 6.1.6, compared with the progenitor cell line T5-1. Second, a DNA-protein complex, termed X-A, formed by nuclear extracts from T5-1 with DRA sequences containing the X box and a few bases 5' and 3' to it, is missing with extracts from 6.1.6. Extracts from some but not all patients with class II-negative immunodeficiency also fail to form X-A, whereas extracts from class II-negative mutants derived from the Burkitt's line Raji do form an apparently normal X-A complex. The X-A complex contains proteins of approximately 22, 32, 82, and 92 kDa that can be cross-linked to a 5-bromodeoxyuridine-substituted X box probe by UV light. A defect in an X box-binding protein, or in a factor required for its binding, is a likely cause for the loss of transcription of the class II genes in 6.1.6.

Cell Line↗

PubMed1991

Characterization of the promoter region of the c-yes proto-oncogene: the importance of the GC boxes on its promoter activity.

In this study we cloned the 5' flanking sequence of the human c-yes gene and identified its promoter region. A 0.53 kilobase pair (kbp) fragment containing the 5' terminus of the c-yes gene showed strong promoter activity when placed upstream of the bacterial chloramphenicol acetyltransferase (CAT) gene and transfected into monkey CV-1 cells. By nuclease S1 mapping multiple transcriptional start sites were detected within the promoter region. Nucleotide sequence analysis revealed that the c-yes promoter region had high G + C contents (64%) and contained six GC box-like sequences (one at the 5' distal region and five in a cluster at the 5' proximal region), but not a TATA box. These features of the c-yes promoter region are similar to those of other protooncogenes, ras-family genes and c-raf-1, and some house-keeping genes. Deletion analysis suggested that the most downstream 0.21 kbp region is primarily important for the promoter activity. This 0.21 kbp region contains one major and another minor transcriptional start site. Five GC box-like sequences were located within this region, and four of them were shown to bind with purified Sp1 transcription factor. Furthermore, using the base-substituted mutants of the Sp1-binding sites, each GC box in the cluster (GC1 to GC4) was shown to affect the c-yes gene expression.

Animals↗

PubMed1990

Regulation of expression of the dnaA gene in Escherichia coli: role of the two promoters and the DnaA box.

The dnaA gene of Escherichia coli specifies a product that is a key element in the initiation of DNA replication. Expression of dnaA occurs from two promoters, 1P and 2P, which flank a DnaA protein binding site (DnaA box). In this paper we describe the effects of DnaA box mutations on transcription from the two promoters, measured with the use of dnaA-galK gene fusions. All of the mutations examined led to a five-fold increase in expression from the upstream promoter, 1P, suggesting that expression from this promoter is negatively autoregulated by binding of DnaA protein to the box. The same mutations led to a decrease in expression from promoter 2P, suggesting that the DnaA box plays a role in activating promoter 2P under normal conditions. Overproduction of DnaA protein, above normal physiological levels, led to decreased transcription from both promoters. We present evidence from analysis of both dnaA mRNA and DnaA protein that promoter 2P is subject to growth rate-dependent control and that this promoter, but not promoter 1P, is completely shut off in the stationary phase of growth.

Bacterial Proteins↗

PubMed1988

A mouse pluripotent embryonal stem cell line stage-specifically regulates expression of homeo-box containing DNA sequences during differentiation in vitro.

Mouse embryonal stem (ES) cells have been shown to provide a new model system suitable for the analysis of different aspects of murine development. This report gives evidence that ES cell lines are also most useful for the study of developmentally regulated gene expression in vitro. Homeo-box containing genes which are suggested to play a key role in the regulation of differentiation steps occurring during embryogenesis are stage-specifically transcribed in differentiating murine ES cells: (i) A mouse embryonal stem cell line (ES-12957) was isolated and characterized with respect to its differentiation potential. When injected subcutaneously into syngeneic mice, ES-12957 cells formed fully differentiated teratomas representing derivatives of all three germ layers. When allowed to grow in suspension cultures in vitro, the cells followed a reproducible developmental pathway forming complex organized 'embryoid bodies' which resembled mouse early postimplantation embryos. (ii) A mouse DNA sequence with homeo-box homology (MH-121) was isolated and structurally analyzed. Transcription of a 1.7 kb RNA species from this DNA sequence was demonstrated in ES-12957 cells which were differentiated in vitro. A second, previously described homeo-box gene (Mo-10) was also shown to be expressed in ES-12957 cells in a stage-specific manner. A 4-kb transcript could be identified exclusively in RNA of cells which were allowed to differentiate for 9 days. These findings support the suggestion that the homeo-box genes of mammals, like those of Drosophila, may have important functions during embryonic development.

Animals↗

PubMed1987

Spatial regulation of homeo box gene expression in Drosophila.

1. Each of the 17 known homeo box genes shows a unique pattern of expression during early embryonic development. Thus, virtually every embryonic cell contains a unique combination of active and inactive homeo box genes. 2. These different permutations of homeo box gene expression are thought to specify unique positional values, and appear to play key roles in selecting diverse pathways of morphogenesis. 3. It has been proposed that selective patterns of homeo box gene expression involve cross-regulatory interactions. For example, it appears that homeotic genes expressed in posterior regions of the embryo (such as abd-A and Abd-B) repress the expression of those homeotic genes expressed in more anterior regions (such as Antp and Ubx). Such interactions might involve sequence-specific DNA binding and occur at the level of transcription. 4. The establishment of selective patterns of pair-rule gene expression along the anterior-posterior embryonic body axis might involve the differential regulation of these genes by a common set of transcription factors. In particular, it appears that eve and ftz show opposite responses to regulatory products that are either directly or indirectly specified by several gap genes.

Amino Acid Sequence↗

PubMed1995

Point mutations in the conserved box 1 region inactivate the human granulocyte colony-stimulating factor receptor for growth signal transduction and tyrosine phosphorylation of p75c-rel.

The human granulocyte colony-stimulating factor receptor (hG-CSFR) belongs to the cytokine receptor superfamily. As with other members of this family, the cytoplasmic domain of hG-CSFR lacks intrinsic tyrosine kinase activity. To identify critical regions mediating growth signal transduction by hG-CSFR, deletions or site-directed amino acid substitutions were introduced into the cytoplasmic domain of hG-CSFR, and the mutant cDNAs were transfected into the murine interleukin-3 (IL-3)-dependent Ba/F3 and FDCP cell lines. Truncation of the carboxy-terminal end of the receptor to the membrane-proximal 53 amino acids of the cytoplasmic domain, which retained the conserved Box 1 and Box 2 sequence motifs, decreased the ability of hG-CSFR to transduce G-CSF-mediated growth signals without an associated loss in receptor binding affinity. Substitution of proline by alanine at amino acid positions 639 and 641 within Box 1 completely abolished the G-CSF-mediated growth signal. Rapid induction of tyrosine phosphorylation of several cellular proteins, including a 75-kD protein (p75) identified as c-rel, was an early event associated with transduction of proliferative signals by hG-CSFR in Ba/F3 transfectants. Mutant receptors containing Pro-to-Ala substitutions that inactivated the receptor for mitogenic activity also inactivated the receptor for tyrosine-specific phosphorylation of p75. These results show that the conserved Box 1 sequence motif (amino acids 634 to 641) is critical for mitogenesis and activation of cellular tyrosine kinases by hG-CSFR.

Amino Acid Sequence↗

PubMed1994

Acquisition and extinction of jumping, two-way shuttle-box and bar press avoidance responses in malnourished rats: effects of shock intensity.

1. In order to investigate the role of avoidance response and shock intensity in avoidance learning in malnourished rats, three avoidance responses (jumping, two-way shuttle-box and bar press) and three shock intensities (0.4, 0.6 and 1.0 mA) were used. Independent groups of 6 rats were used for each response topography and shock intensity. 2. Malnourished male Wistar rats were suckled by mothers fed a 12% casein diet during the lactation period (0-21 days of age) while the mothers of well-nourished controls received a 25% casein diet. After weaning (21st day), all animals received a commercial lab chow diet until 70 days of age, when the avoidance training started. 3. Malnutrition did not affect the acquisition of the avoidance response, but malnourished groups required more trials to extinguish jumping and two-way shuttle-box. During the acquisition phase all animals learned the jump response faster in comparison to bar press and shuttle-box avoidance responses. Both groups in the acquisition phase responded faster with 1.0 mA when compared to lower intensities (0.6 and 0.4 mA). The malnourished animals showed lower latency of avoidance in the jumping response when compared with well-nourished animals. During the extinction phase there was a significant effect of diet, response topography and shock intensity in the latency to respond and trials to criterion. The increased resistance to extinction in malnourished rats was particularly evident with 1.0 mA in the two-way shuttle-box response. 4. These results suggest that contradictory data related to the acquisition of the avoidance response in malnourished animals cannot be attributed to response topography or variations in shock intensity. Furthermore, our results also indicate that resistance to extinction and latency to respond are appropriate parameters for detecting differences between well-nourished and malnourished animals.

Animals↗

PubMed1993

Amalgam retention using pins, boxes, and Amalgambond.

The purpose of this study was to evaluate and compare the retentive strength of amalgam to tooth structure by utilizing pins, boxes, Amalgambond adhesive only, and Amalgambond adhesive plus retentive boxes. There was no statistical difference (P = 0.05) in retentive strength of the amalgam between the group utilizing pins alone and the group utilizing Amalgambond plus retentive boxes. However, both of these sample groups demonstrated statistically greater retentive strengths than either the boxes only or Amalgambond only groups.

Analysis of Variance↗

PubMed1994

Box plate fixation of the symphysis pubis: biomechanical evaluation of a new technique.

The purpose of this study was to compare common techniques of pubic symphyseal fixation with a new method, the "box plate," for fractures of the pelvis where the bone is osteopenic. This symphyseal fixation construct consists of two, two-hole, 4.5-mm narrow dynamic compression plates (DCP) oriented parallel to one another. One plate is recessed within the symphysis, and the other is located on the pubic tubercles. The plates are interlocked using two 6.5-mm fully threaded screws, forming a box-like construct. To determine the mechanical properties of this construct, five fresh, cadaveric pelvic specimens with a mean age of 75 years were harvested. The femora of each specimen were potted into containers and fixed to the base of a materials testing machine. The pelvis was constrained from rotating about the hip joints by anterior and posterior restraints. A vertical compressive load was applied through the lumbar spine. Force to a magnitude of 1,000 N was applied through three cycles. Gapping motions at the symphysis pubis (SP) and the sacroiliac (SI) joints, and flexion-extension of the sacrum with respect to the ilia were measured under the following conditions: (a) intact, (b) SP ligament, unilateral anterior SI ligaments, and ipsilateral sacrospinous and sacrotuberous ligaments disrupted (anteroposterior compression type II injury), and these injuries fixed using (c) a 4.5-mm narrow two-hole DC plate placed on the superior SP held by two cancellous bone screws, (d) the DC plate well as a single 7.0-mm cannulated cancellouoffliosacral lag screw across the injured SI joint, (e) the DC plate and a five-hole 3.5-mm reconstruction plate on the anterior SP, (f) a 3.5-mm, four-hole, DC plate on the superior SP using four fully threaded screws, and (g) the box plate symphyseal construct described above. All fixations reduced SP joint gapping compared to the disrupted joint. However, all but the box plate still allowed significantly greater motion than the intact SP joint. No fixation significantly reduced SI joint gapping or sacral flexion compared to the injured state.

Aged↗