Transformation of normal rat kidney cells by v-K-ras enhances expression of transin 2 and an S-100-related calcium-binding protein.

v-K-ras transformants of normal rat kidney cells (KNRK) exhibit cell surface-related, transformation-specific properties, including cell-surface fibronectin depletion, induction of anchorage- and density-independent growth, and increased synthesis of transforming growth factors alpha and beta. To search for potential distal effectors of v-K-ras-mediated transformation, we prepared a rabbit antiserum directed against intact KNRK cells to immunoprecipitate and compare proteins from detergent lysates and conditioned media of labeled NRK, KNRK, B77-NRK (a v-src transformant) and ts-371-NRK cells (a Ki-MSV encoding a temperature-sensitive p21v-K-ras). Proteins with enhanced expression in both wild-type v-K-ras and v-src transformants included a cell-surface phosphoglycoprotein with apparent Mr of 79,000 (79K) modified from an 85K protein observed in NRK cell lysates, a cytoplasmic 47K and a 10K protein, and a 57K secreted glycoprotein. A KNRK-specific 21K membrane-associated protein and secreted 59K and 36K secreted glycoproteins were also detected. The expression of the 36K and 59K proteins best correlated with temperature-dependent activation of the ts-371-NRK p21v-K-ras. Immunoselection of recombinant clones from a KNRK-specific lambda gt11 cDNA library allowed identification of the 59K and 10K proteins as transin 2 and an S-100-related calcium-binding protein identified as p9Ka/42A but not previously associated with oncogenic transformation of rat cells. Transin 2 detection by a cell-derived antiserum may also suggest the presence of specific cell-surface binding sites for this enzyme.

[Record filing in a department of radiology. Value of a microcomputer]. / L'archivage dans un service de radiologie. Intérêt d'un micro-ordinateur.

Advantages of computer-based record filing when compared with conventional methods are outlined, and a detailed description given of the use of a microcomputer for establishing a filing system in a department of radiology.

[Osteonecrosis of the femoral head of sickle-cell origin. Apropos of 19 cases observed during a year in Benin. Proposal for radiologic classification]. / Les ostéonécroses de la tête fémorale d'origine drépanocytaire. A propos de 19 cas rencontrés en un an au Bénin. Proposition de classification radiologique.

The author reports 19 aseptic necrosis of the femoral head, due to sickle-cell anemia, collected in 14 black people native from Benin. Sickle-cell anemia is the most frequent etiology of osteonecrosis there. They are generally observed between 10 and 30 years of age. A systematic and complete study of the radiological signs has been carried out. The peculiar severity of the disease in black Africa is related to the chronicity of the sickling disorder, the delay brought to the diagnostic, the lack in therapeutic media. A six stage X-ray classification is argued because of the most severe aspects.

[Feasibility of radiological exploration of the main bile ducts for stones (author's transl)]. / A propos de la fiabilité des explorations radiographiques de la voie biliaire principale pour lithiase.

The results of cholangiography conducted pre-and post operatively (through Tatute), and during choledochotomy on 100 patients for confirmed or suspected bile duct stones are analyzed. Pre-operative opacification of the main bile ducts resulted in 13 p. cent of false positives and in a similar number of false negatives. Results of investigation during operation produced similar errors respectively in 14 and 2p. cent o of cases. Several recommendations are outlined to limit the proportion of unnecessary choledochotomies without increasing the number of neglected stones.

[Roentgenographic features in pigmented villonodular synovitis (concerning a carpal location) (author's transl)]. / L'image radiologique de la synovite villonodulaire hémopigmentée. A propos d'une localisation carpienne.

Pigmented villonodular synovitis is a benign proliferation of the synovial membrane. Its pathogenesis is not well known. It results in an articular swelling without great pain. Its roentgenographic aspect is a combination of an opacity of soft tissue and epiphyseal damages, i.e. cortical erosions and lacunae surrounded by osteosclerosis. Hyperplasty of the synovial membrane is explicited by arthrography and arteriography. The authors report one case of pigmented villonodular synovitis in the carpus: that is an uncommon location of that disease which most often involves the knee.

CT findings of atypical forms of phakomatosis.

Three cases with unusual manifestations of phakomatosis are reported. The first two had clinical symptoms of neurofibromatosis but CT disclosed nodular subependymal calcifications as in tuberous sclerosis. The third one presented with cerebral calcifications as found in both tuberous sclerosis and Sturge-Weber syndrome, though he had no clinical symptoms of phakomatosis.

[Osteonecrosis of the femur head caused by sickle cell anemia in Benin. Epidemiologic and radiologic aspects]. / Les ostéonécroses de la tête fémorale d'origine drépanocytaire au Bénin. Aspects épidémiologiques et radiologiques.

The author reports 19 aseptic necrosis of the femoral head, due to sickle cell anemia, collected in 14 black people native from Benin. Sickle cell anemia is the most frequent etiology of osteonecrosis there. They are generally observed between 10 and 30 years of age. A systematic and complete study of the radiological signs has been carried out. The peculiar severity of the disease in black Africa is related to the chronicity of the sickling disorder, the delay brought to the diagnostic, the lack in therapeutic media.

Differential processing of osteopontin transcripts in rat kidney- and osteoblast-derived cell lines.

Using immunoprecipitation and tryptic peptide microsequencing we confirmed the identity of normal rat kidney (NRK) cell-secreted 69-kDa major phosphoprotein as osteopontin (OP). We then immunoselected a 1.4-kilobase pair (kb) OP cDNA from a lambda gt11 library prepared from Kirsten sarcoma virus-transformed NRK (KNRK) cellular mRNA, using rabbit anti-69-kDa OP serum. Sequence analysis of this cDNA revealed the presence of a 52-nucleotide-long insert in the 5'-noncoding region, which was absent in OP cDNA cloned from the cDNA library of ROS 17/2.8 rat osteosarcoma cells. The insert sequence is flanked by putative intron splice junctions and is located 15-nucleotide upstream of the translational initiation site. An insert-specific 30-mer oligonucleotide probe hybridized to a single 1.5-kb RNA species from both NRK and KNRK cells, but not from ROS 17/2.8 cells. However, Southern analysis showed the presence of this insert sequence in the genomic DNA of both NRK and ROS 17/2.8 cells. Furthermore, PCR amplification of the insert-containing region using genomic DNAs from both NRK and ROS 17/2.8 cells gave products of identical size and sequence. Since OP is a single copy gene, these data provide strong evidence for differential cell type-specific processing of OP transcripts. In addition, we demonstrate that, in contrast to most transformed cells, levels of OP expression are significantly reduced in KNRK cells as compared to NRK cells.

Dibutyryl cAMP inhibits expression of transformation-related properties in Kirsten murine sarcoma virus transformed Balb/c-3T3 cells despite continued presence of p21v-Ki-ras.

We studied the effects of simultaneous treatment with 0.1 mM N6, O2'-dibutyryl cAMP (dbcAMP) and 1 mM theophylline on several transformation-specific properties and on levels of the Kirsten murine sarcoma virus (Ki-MSV) transforming gene product p21v-Ki-ras, in a Ki-MSV-transformed mouse cell line (Balb/c-3T3, clone A31; KA31). The rate of logarithmic growth, cell motility, and final saturation density were reduced in dbcAMP-treated KA31 cultures. Capabilities for anchorage-independent growth were reduced in treated cells, to levels similar to those observed for the untransformed parental A31 cell line. Treatment with dbcAMP had no observable effect on the binding of 125I-labeled epidermal growth factor and did not alter fluorescence staining patterns for actin microfilaments and fibronectin which, although characteristic of normal cells, were also present in KA31 cells. Changes induced by dbcAMP were readily reversible, except for loss of anchorage-independent growth. However, this property was also reversible, provided removal of dbcAMP occurred 48 h prior to inoculation into soft agar medium. Immunoprecipitation with a monoclonal antibody directed against the protein p21v-Ki-ras (Y13-259) revealed the continued presence of this protein in dbcAMP-treated KA31 cells. We, therefore, conclude that cAMP mediates the inhibition of growth-related transformation-specific properties either by acting at steps subsequent to the expression of p21v-Ki-ras or on a pathway independent of p21ras function.

An allergenic polypeptide representing a variable region of hsp 70 cloned from a cDNA library of Cladosporium herbarum.

BACKGROUND: Extracts of Cladosporium herbarum, a major source of fungal aeroallergens, exhibit a complex profile of IgE-binding proteins. Yields of conventionally purified allergens from this mold have been insufficient to permit further molecular analyses. OBJECTIVE: To enhance and simplify the purification of allergens from C. herbarum, we have sought to use recombinant DNA techniques to clone, identify and bacterially express immunoselected C. herbarum allergens. METHODS: We constructed a cDNA library in lambda ZAP II using mRNA isolated from C. herbarum. From this library, phage clones encoding a new allergen were immunoselected using pooled human atopic IgE. The cloned cDNA was excised from the phage vector as a recombinant pBluescript II SK-phagemid and sequenced. Expression of the recombinant allergen was carried out in E. coli XL1-blue transformants of the phagemid. Bacterial lysates from cells induced to express the cloned allergen were immunoblotted and probed with individual human atopic IgEs. RESULTS: The cDNA clone encodes a 278 amino acid polypeptide homologous to the C-terminal portion of 70 kDa heat shock protein (hsp 70). The polypeptide possesses features common to other hsps 70, i.e. a similar hydropathic profile and a variable C-terminal region with conserved sequence at the very C-terminus. Binding of the recombinant peptide to IgE from 38% of atopic sera or plasma from individuals allergic to C. herbarum was demonstrated. CONCLUSION: These results indicate that amino acid substitutions are relatively conserved even in the variable C-terminal regions of hsp 70 species. Thus, this study should draw attention to the possibility of induction of anaphylactic responses in a sensitized individual when hsp 70 from any pathogenic species is administered for vaccination.

[Ultrasonic diagnosis of acute cholecystitis. Critical study of 50 verified cases]. / Le diagnostic ultrasonore de la cholécystite aiguë. Etude critique de cinquante observations contrôlées.

A retrospective study of fifty surgically proven cases of acute cholecystitis and a review of the literature enabled the authors to summarise the sonographic features of the disease. These may include one or several of the following signs: thick gall bladder wall with an occasional posterior hypoechogenic rim, diffuse echogenicity of the purulent bile and local tenderness at the passage of the transducer. If an abscess is present, there is a hypoechogenic area in contact with the gall bladder. The technical problems of the B scan performed on an emergency basis make the use of real time intercostal scans invaluable. Ultrasound should be the first and may be the radiological examination performed in this situation.

Multiple sclerosis and corpus callosum atrophy: relationship of MRI findings to clinical data.

Among 110 patients (45 men, 65 women), aged 15 to 66, with clinical and/or biological diagnosis of multiple sclerosis (MS), severe to moderate corpus callosum (CC) atrophy was observed in 67 (60%) patients. Correlation between CC atrophy, brain atrophy, duration and severity of clinical symptoms, and high signal white matter areas, was carried out in 90 patients. Mean age was 46 years for patients with severe CC atrophy, and 33 years for those without atrophy. Mean duration of the disease was 14 years in patients with severe atrophy, and 5 years in patients without atrophy. Severity of clinical symptoms is more pronounced in patients with severe CC atrophy. Numerous or large white matter high signal areas are observed in patients with severe CC atrophy on T2-weighted images. CC atrophy appears earlier than brain atrophy in the course of MS.

N-terminus of a major allergen, Alt a I, of Alternaria alternata defined to be an epitope.

A 20mer peptide representing the N-terminus of a major allergen, Alt a I, of Alternaria alternata was synthesized and examined for its antibody binding and antibody induction activities. Alt a I peptide-BSA conjugate reacted with both human IgE and rabbit anti-Alt a I IgG in ELISA, albeit the binding of the peptide to IgE was relatively weak. Control conjugate showed no antibody binding. These results indicated that the N-terminus of Alt a I is an antibody binding site. Moreover, peptide-KLH conjugate and nonconjugated peptide induced both IgG and IgE antibodies in Balb/c mice that recognized both native Alt a I allergen and peptide-BSA conjugate. Since the free peptide was able to induce antibodies in vivo, the peptide may also possess a T cell epitope.

Molecular cloning of IgE-binding fragments of Alternaria alternata allergens.

Exposure to the hyphomycete Alternaria alternata is recognized as an important risk factor for asthma. IgE immunoblotting has been used to catalogue the number and Mr of allergens in A. alternata extracts, with estimates ranging from 10 to 30, although few are present in nearly all extracts studied. Several A. alternata allergens have been cloned, including a subunit of the major allergen Alt a 1, ribosomal P2 phosphoprotein, aldehyde dehydrogenase and a yeast YCP4 homolog. We have cloned two sequences encoding IgE-binding fragments of allergens from an A. alternata lambda gt11 cDNA library using pooled atopic sera from A. alternata-sensitive individuals. One is homologous to a region near the C-terminus of hsp70 from Cladosporium herbarum; a near-complete isoallergen variant of A. alternata ribosomal P2 protein was also cloned. Their lacZ fusion proteins had reactivities of 5 and 14%, respectively, with individual atopic sera, indicating that the corresponding allergens are both minor. This study describes one new A. alternata allergen candidate and implicates ribosomal P2 protein as an allergen thtat is stable between independently isolated clones.

Isolation and expression of a cDNA clone encoding an Alternaria alternata Alt a 1 subunit.

Alternaria alternata is recognized as an important source of fungal aeroallergens. Alt a 1, the major allergen of this mold, is a dimer of disulfide-linked subunits that migrate in SDS-PAGE under reducing conditions at apparent M(r)s of 14,500 and 16,000. IgE antibodies to this protein are present in the sera of >90% of A. alternata-sensitive individuals. Previous studies from this laboratory showed that the N-termini twenty amino acids of the purified subunits are nearly identical. We now report the isolation of clones from an A. alternata (strain 34-016) cDNA library constructed in lambda(gt)11, using rabbit IgG antiserum against partially purified Alt a 1. One of nineteen clones selected from screens totalling 305,000 pfu (rb51) was sequenced, and determined to harbor an insert of 660 bp. An in-frame open reading frame within the cloned insert encodes a peptide of M(r) 16,960 that bears no significant homology to known allergens or proteins. The size of the rb51 transcript was determined to be approximately 0.7 kb by Northern analysis of A. alternata total RNA. The largely hydrophobic N-terminal region of the peptide contains an alpha-helical domain and other features characteristic of membrane targeting or secretory signals. The peptide sequence downstream of this region matches previously sequenced Alt a 1 N-terminal from two independent sources at 17 of 20, and 24 of 26 positions. Recombinant Alt a 1 expressed as a secreted protein in Pichia pastoris exists as a dimer in conditioned medium, as shown by immunoblotting under nonreducing conditions. Recombinant Alt a 1, like the natural allergen in A. alternata extracts, is also reactive with serum IgE from A. alternata-sensitive individuals.