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Biomedical subjects

J Sprague

Publications and source records attributed to J Sprague.

15 recordsLinked to original sources

Interleukin-12 p40 m-RNA expression in human kidney allograft biopsies.

Interleukin-12 (IL-12) is a heterodimeric cytokine implicated in the early differentiation of naive T-lymphocytes into the Th1 subset. IL-12 is important for induction of the cellular immune response against viruses, intracellular parasites and neoplasms. Its role in alloresponsiveness has not been fully elucidated. Preliminary data in the literature point toward the prevalence of Th1 lymphocytes in processes of allograft rejection. In attempt to further investigate the expression of this cytokine during episodes of cellular rejection of renal allografts, we searched for IL-12 message in human kidney allograft biopsies using the reverse transcriptase-polymerase chain reaction technique. Twenty-three allograft core biopsies from 19 patients were obtained percutaneously for clinical indications in 18 cases, and as part of an investigational protocol in five cases. A portion of the tissue was used for RNA extraction using the guanidium-thiocyanide phenol-chloroform method. Histology was performed on the remaining core material. Ten mg of total RNA were used for reverse transcription. PCR of the c-DNAs was done for 40 cycles using primers for the p40 subunit of IL-12 and GAPDH which was used as a control. PCR products were photographed after electrophoresis, transferred to a nylon membrane and hybridized with a radiolabelled cloned human IL-12 p40 1 kb c-DNA fragment. Autoradiographies were developed after 20-min exposure. All samples were run in triplicate. IL-12 p40 m-RNA was expressed in all 17 biopsies showing acute cellular rejection as well as in all three biopsies showing focal interstitial fibrosis. No message was found in the presence of normal allograft histology. This is the first in vivo report of IL-12 p40 subunit m-RNA expression during renal allograft rejection in humans. The role of this Th1 cytokine in the alloresponse deserves further investigation.

Adult↗

Building blocks. Roundtable discussion.

Former Hospitals & Health Networks executive editor Mark Hagland and Health Facilities Management editor Kurt Luchs and associate editor Catherine Quayle recently hosted a roundtable discussion on design and construction issues in health care. They wanted to know what health care senior executives and experts in the design and construction field thought about today's most pressing issues and biggest trends. Hagland and Luchs assembled a group or experts and executives in Chicago and set the tape rolling. Here are excerpts from the discussion.

Decision Making, Organizational↗

Impact of massive doses of vitamin A on nutritional blindness in Bangladesh.

Impact of 6-monthly massive dosings of preschool-age children with oral vitamin A (VAC: 200,000 IU of oil soluble retinyl palmitate with 40 IU vitamin E) was evaluated in Bangladesh. In 100 sites, 11,889 households were visited and eyes of 22,335 children aged 3-71 mo were examined. About half the rural target population and less than 20% urban slum population were being reached. Risk of night blindness was halved for children reportedly given VAC, although 2.5% of the reportedly protected population were still night blind. There was no significant reduction in prevalence of Bitot's spot. Risk of corneal ulcers or keratomalacia (X3A/B) was 2.7 times higher in children not given VAC. Based on reported coverage, efficacy of protection against potentially blinding corneal lesions was 63%. For maximum impact on eye lesions, massive dosing with vitamin A at ideally less than 6-monthly intervals needs to be combined with other nutrition and health interventions.

Bangladesh↗

Landholding, wealth and risk of blinding malnutrition in rural Bangladeshi households.

The 1982-1983 Bangladesh nutritional blindness study visited 11,618 rural households and examined 18,660 preschool-age children in an effort to determine the prevalence and determinants of eye lesions and loss of sight due to vitamin A deficiency (xerophthalmia). Risk of xerophthalmia was significantly higher for children from households without any of the indicators of relative wealth used. Almost 80% of blind children from landless households, and even a very small garden reduced considerably the chances of a household having a xerophthalmic child. Poorer households with access to less than 0.3 acres land or no garden or without a tin roof, wristwatch, radio or cycle were at least twice as likely as their more fortunate neighbours to have a young child with any type of xerophthalmia. Taking account of such socio-environmental risk factor weightings would direct the scarce resources of intervention programmes to households and children who most need them.

Bangladesh↗

Characterization of mouse myelin basic protein messenger RNAs with a myelin basic protein cDNA clone.

Using a family of synthetic tetradecamer oligonucleotides as a primer for cDNA synthesis and a second family of tetradecamers as a hybridization probe, we have prepared and isolated a cDNA clone of mouse myelin basic protein (MBP). The clone, pNZ111, corresponds to the region of the mRNA that codes for an amino acid sequence present in all four major forms of MBP. The relative abundance of MBP mRNA, estimated by dot blot hybridization, increased with the age of the mouse to a maximum at 18 days, then decreased to about one-fourth of that amount at later ages. Mouse MBP mRNAs, selected by their ability to hybridize to the clone, translate into the four forms of myelin basic protein. In RNA blot analyses, pNZ111 hybridized to multiple species of mouse mRNA. The predominant hybridization is to a broad band of RNAs ranging in length from 2,350 to 2,100 bases. These mRNA species are extremely long, considering that the largest MBP could be encoded by approximately 600 bases. In addition to these, there are also minor bands that hybridize with pNZ111, including a band of 4,100 bases and smaller ones of 1,900, 1,500, and 1,200 bases.

Amino Acid Sequence↗

Expression of a recombinant DNA gene coding for the vesicular stomatitis virus nucleocapsid protein.

A cDNA clone containing the entire vesicular stomatitis virus nucleocapsid gene was assembled by fusing portions of two partial clones. When the cDNA clone was inserted into a new general-purpose eucaryotic expression vector and introduced into appropriate host cells, abundant N-protein synthesis ensued. The expressed protein was indistinguishable from authentic N protein produced during vesicular stomatitis virus infections. The recombinant N protein was recognized by a polyclonal antibody and two different monoclonal antibodies and could not be resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis from authentic N. Our results suggest that the recombinant N protein produced in transfected cells rapidly aggregates into high-molecular-weight complexes in the absence of vesicular stomatitis virus genomic RNA.

Capsid↗

In vitro transcription of vesicular stomatitis virus: initiation with GTP at a specific site within the N cistron.

In vitro transcripts of vesicular stomatitis virus (VSV) were either 5'-terminally labeled by incorporation of [beta-(32)P]GTP or were selected on Hg-agarose after incorporation of gamma-thio-GTP. Capped RNAs ranged in size from 23 nucleotides, the shortest capped RNA detected, to full-length message size. The 5'-terminal sequences corresponded to those of N message and to a small amount of NS message. Approximately 14% of the capped N gene transcripts were terminated at positions 86 to 90 of the VSV genome, giving rise to specific, 36 to 40-nucleotide-long, capped RNA species. The GTP-initiated RNAs were short with a predominant 28-nucleotide-long RNA species. A minor portion was as large as mRNAs. Nucleotide sequence analyses of the short RNA revealed that it was specifically initiated at positon 91 of the VSV genome, 41 nucleotides within the N cistron. This corresponds exactly to the site where transcription of the 40-nucleotide-long, capped RNA terminated. Initiation with GTP at position 91 occurred at approximately the same frequency as termination of the capped RNA at position 90, suggesting that intracistronic initiation at position 91 may depend upon termination of transcription of the 5'-proximal region and therefore may be sequential. This unique RNA represents the first transcript of VSV which was initiated at an intracistronic site with GTP, and may also represent the first example of a transcript derived from a stop/start mechanism of VSV transcription in vitro. Although initiation occurred frequently at the beginning of the N cistron yielding 11 to 14-nucleotide-long, [beta-(32)P]ATP-labeled transcripts (D. F. Pinney and S. U. Emerson, J. Virol. 42:889-896, 1982), capping of these short RNAs was not detected. This suggests that transcripts may have to be 15 to 23 nucleotides long to be accepted as substrates by the guanyltransferase.

Base Sequence↗

Cell-free synthesis of measles virus proteins.

Polyadenylated mRNA extracted from cytoplasm of measles virus-infected Vero cells was translated in a cell-free system. Three of the polypeptides obtained corresponded to nucleocapsid protein, phosphoprotein, and membrane protein of measles virions. A fourth polypeptide, present in measles virus-infected cells, could be generated by addition of Vero cytoplasmic extract and was identified as a cleavage product of the nucleocapsid protein.

Capsid↗

The effect of noncontingent sensory reinforcement, contingent sensory reinforcement, and response interruption on stereotypical and self-injurious behavior.

Three analyses were conducted to assess the effects of different consequent stimuli on the rate of stereotypical and self-injurious behavior performed by two individuals with severe developmental disabilities and dual sensory impairments. An analogue functional analysis documented an undifferentiated pattern of problem behavior across all conditions for Participant 1. Data for Participant 2 indicated an undifferentiated pattern with lower frequencies in the demand condition. Stimuli chosen to compete with the type of sensory stimulation produced by the stereotypy and self-injurious behavior were presented noncontingently during play conditions. Noncontingent presentation of the specially selected stimuli resulted in reductions in stereotypy and self-injurious behavior. Finally, contingent presentation of the same stimuli with and without response interruption was assessed in a demand context. Contingent presentation of the specially selected stimuli plus response interruption resulted in more suppression than contingent sensory stimulus presentation alone. Results are discussed as to competing and concurrent schedules of reinforcement.

Adult↗

Cast syndrome: the superior mesenteric artery syndrome.

Cast syndrome, clinically known as superior mesenteric artery syndrome (SMAS), is gastric dilatation with partial or complete obstruction of the duodenum. Although rare, it is most frequently seen in orthopaedic patients who have had spinal surgery or who are in hip spica or body casts. Obstruction occurs when there is compression of the duodenum between the superior mesenteric artery anteriorly and the aorta and spinal column posteriorly. Obstruction can occur within days of surgery or casting or may not develop for several weeks. Treatment for SMAS varies from conservative nonoperative to operative procedures. Complications can be severe if symptoms are not quickly recognized and treatment instituted in a timely manner.

Diagnosis, Differential↗