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Biomedical subjects

J B Joshi

Publications and source records attributed to J B Joshi.

15 recordsLinked to original sources

Molecular cloning and chromosomal mapping of the mouse cyclin-dependent kinase 5 gene.

Cyclin-dependent kinase 5 (Cdk5) is predominantly expressed in neurons. In vitro, Cdk5 purified from the nervous tissue phosphorylates both high-molecular-weight neurofilament and microtubule-associated tau. The mouse gene encoding Cdk5 (Cdk5) was found to be 5 kb in length and divided into 12 exons. All of the exon-intron junctions matched the expected consensus sequence with the exception of the splice junction for intron 9, which has AT and AC dinucleotides instead of the usual GT and AG bordering sequence. In the 5'-flanking region of mouse Cdk5, several putative promoter elements were present, including AP1, Sp1, PuF, and TATA motifs. A metal regulatory element was also identified at position -207 to -201. Nucleotide sequence analysis of mouse Cdk5 showed high identity to the homologues of other vertebrate species, indicating that this kinase is highly conserved during evolution. Mouse Cdk5 was mapped to the centromeric region of mouse chromosome 5.

Amino Acid Sequence

Transactivation of the proenkephalin gene promoter by the Tax1 protein of human T-cell lymphotropic virus type I.

Human T-cell lymphotropic virus type I (HTLV-I), an etiologic agent for adult T-cell leukemia, is strongly associated with certain neurological diseases. The HTLV-I genome encodes a protein, Tax1, that transactivates viral gene transcription. CD4-positive T helper lymphocytes express the proenkephalin gene, and enkephalins have been implicated as neuroimmunomodulators. We have investigated the effect of Tax1 on the proenkephalin gene promoter in C6 rat glioma cells and demonstrated its transactivation. Analysis using 5' deletion mutants of the promoter region showed that sequences upstream of base pair -190 are necessary for maximal transactivation. Forskolin, a cAMP modulator, synergistically increased Tax1-mediated transactivation of the proenkephalin promoter. Neither Tax1 transactivation alone nor Tax1/cAMP synergism exclusively involved cAMP-responsive elements. Endogenous proenkephalin gene expression increased in Tax1-expressing C6 cells. Since HTLV-I infects lymphocytes, which express proenkephalin mRNA, Tax1 transregulation of proenkephalin expression may provide bidirectional communication between the nervous and immune systems in HTLV-I-related diseases.

Animals

Continuous counter-current two-phase aqueous extraction.

Continuous counter-current column operation provides operating convenience for contacting two-phase aqueous partitioning systems for protein extraction. We discuss in detail the important parameters for designing spray, packed, plate and York-Scheibel columns for protein recovery using both polymer-polymer and polymer-salt two-phase aqueous systems. We compare the various contractors for their operating and extraction efficiency. The work also provides a step-by-step design procedure and specific recommendations for future data needs.

Methods

Influence of the mode of preparation on the distortion and consistency of homologous costal cartilage implants.

Nasal implants of autologous costal cartilage have an undesirable tendency to distortion. This tendency may be reduced by the process of devitalization, which occurs in the preparation of homologous costal cartilage for storage in a tissue bank. An in vitro study has been undertaken, utilizing five common methods of cartilage preparation (cialit, ethyl alcohol, formalin, glutaraldehyde and irradiation) to determine whether there is any significant variation in the degree of distortion which occurred and any variation in cartilage consistency. We conclude that all of the methods evaluated would produce banked homologous costal cartilage with an equally low incidence of distortion but implants are less readily carved from cartilage stored in cialit.

Cartilage

Correlation between nuclear glucocorticoid receptor levels and casein gene expression in murine mammary gland in vitro.

The relationship between nuclear binding of glucocorticoid-receptor complex and casein gene expression was studied in organ culture of the whole mammary gland of the mouse. Pyridoxal 5'-phosphate was used as a modulatory agent for measuring nuclear binding of the receptor complex. Addition of 2 mM and 5mM pyridoxal-5'-P in the medium (Waymouth's MB752/1) resulted in 4- and 12-fold increase of its concentration in the glands incubated with insulin, prolactin, and hydrocortisone. Pyridoxal-5'-P also caused a 52% and 92% inhibition of nuclear binding of [3H]dexamethasone in the glands at 2 mM and 5 mM concentration in the presence of the same hormones in the medium. Corresponding to the reduced nuclear binding of the receptor complex casein mRNA levels, measured by a specific cDNA probe was reduced 86% and over 90% in the glands exposed to 2 mM and 5 mM pyridoxal-5'-P, respectively, in presence of insulin, prolactin, and hydrocortisone in the medium. Withdrawal of pyridoxal-5'-P from the medium restored nuclear binding of the receptor complex near the level of control glands incubated only with the hormones. mRNA casein levels also increased in the gland in the pyridoxal-5'-P-free medium containing the same hormones. This indicates that pyridoxal-5'-P does not alter the specific hormone responsiveness of the mammary cells and its action mediated at the level of the glucocorticoid receptor can influence hormone-inducible expression of the casein genes. Thus, glucocorticoid plays a major role in the multiple hormone regulation of the milk protein gene(s). The findings also suggest that the breast tissue concentration of the vitamin B6 derivative may influence the physiology of lactation in nursing mothers.

Animals