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M Ross

Publications and source records attributed to M Ross.

At least 199 records · Page 11Linked to original sources
PubMed1992

Production and characterization of recombinant insulin-like growth factor-I (IGF-I) and potent analogues of IGF-I, with Gly or Arg substituted for Glu3, following their expression in Escherichia coli as fusion proteins.

The development of an efficient expression system for insulin-like growth factor-I (IGF-I) in Escherichia coli as a fusion protein is described. The fusion protein consists of an N-terminal extension made up of the first 46 amino acids of methionyl porcine GH ([Met1]-pGH) followed by the dipeptide Val-Asn. The latter two residues provide a unique hydroxylamine-sensitive link between [Met1]-pGH(1-46) and the N-terminal Gly of IGF-I. Downstream processing of the fusion proteins involved isolation of inclusion bodies, cleavage at the Asn-Gly bond, refolding of the reduced IGF-I peptide and purification to homogeneity. This expression system was also used to produce two variants of IGF-I in which Glu3 was substituted by either Gly or Arg to give [Gly3]-IGF-I and [Arg3]-IGF-I respectively. Production of milligram quantities of IGF-I peptide was readily achieved. The purity of the IGF-I, [Gly3]-IGF-I and [Arg3]-IGF-I was established by high-performance liquid chromatography and N-terminal sequence analysis. [Gly3]-IGF-I and [Arg3]-IGF-I were more potent than IGF-I in biological assays measuring stimulation of protein synthesis and DNA synthesis or inhibition of protein breakdown in rat L6 myoblasts. Both analogues bound very poorly to bovine IGF-binding protein-2 and slightly less well than IGF-I to the type-1 receptor on rat L6 myoblasts. We conclude that reduced binding to IGF-binding proteins rather than increased receptor binding is the likely explanation for the greater biological potency of the analogues compared with IGF-I.

Amino Acid Sequence↗
PubMed1992

Novel recombinant fusion protein analogues of insulin-like growth factor (IGF)-I indicate the relative importance of IGF-binding protein and receptor binding for enhanced biological potency.

An efficient expression system in Escherichia coli for several biologically active insulin-like growth factor-I (IGF-I) fusion peptide analogues is described. These novel IGF-I fusion protein analogues have properties that make them very useful reagents in the investigation of IGF-I action. The analogues comprise an IGF-I sequence and the first 11 amino acids of methionyl porcine growth hormone (pGH) and include [Met1]-pGH(1-11)-Val-Asn-IGF-I, which contains the authentic IGF-I sequence, and two analogues, [Met1]-pGH(1-11)-Val-Asn-[Gly3]-IGF-I and [Met1]-pGH(1-11)-Val-Asn-[Arg3]-IGF-I, where Glu-3 in the human IGF-I sequence has been replaced by Gly or Arg respectively. The three peptides are referred to as Long IGF-I, Long [Gly3]-IGF-I or Long [Arg3]-IGF-I depending on the IGF-I sequence present. Production of the purified fusion peptides was aided by folding the reduced and denatured fusion peptide sequence under conditions that gave very high yields of biologically active product. Introduction of a hydrophobic N-terminal extension peptide appears to facilitate the correct folding of the IGF-I analogues compared with that obtained previously when folding normal-length IGFs. The biological activities of the IGF-I fusion peptides were compared with authentic IGF-I and the truncated analogue, des(1-3)IGF-I. In L6 rat myoblasts, all the analogues were more potent than authentic IGF-I in their abilities to stimulate protein and DNA synthesis and inhibit protein breakdown. In H35 hepatoma cells, where the IGFs act through the insulin receptor, the Long IGF-I analogues maintained a similar potency relative to IGF-I as was observed in the L6 myoblasts. The order of biological potency in cell lines secreting IGF-binding proteins (IGFBPs) into the medium was Long [Arg3]-IGF-I-des(1-3)IGF-I greater than Long [Gly3]-IGF-I greater than Long IGF-I greater than IGF-I. In chicken embryo fibroblasts, a cell line that does not secrete detectable IGFBPs into the medium, Long [Arg3]-IGF-I, was less potent than IGF-I. Investigation of receptor and IGFBP association by these analogues reinforced our previous findings that N-terminal analogues of IGF-I show increased biological potency due to changes in the degree of their IGFBP interactions.

Amino Acid Sequence↗
PubMed1992

Production and characterization of recombinant chicken insulin-like growth factor-I from Escherichia coli.

Recombinant chicken insulin-like growth factor-I (cIGF-I) has been produced in Escherichia coli after first modifying a plasmid that coded for a human IGF-I (hIGF-I) fusion protein, in order to introduce codons for the eight amino acid substitutions. The cIGF-I fusion protein, deposited in bacterial inclusion bodies, was dissolved under reducing conditions, desalted, subjected to anion-exchange chromatography to remove proteinases, refolded and partially purified by reverse-phase high-performance liquid chromatography. The fusion protein was cleaved with hydroxylamine after which cIGF-I was purified to homogeneity by three additional chromatographic steps. Recombinant cIGF-I was equipotent with hIGF-I in cell culture bioassays of protein synthesis and breakdown using rat L6 myoblasts and chick embryo fibroblasts. Binding of radiolabelled cIGF-I and hIGF-I was also equivalent in the two cell lines, as was their binding in ligand blots of chicken, sheep and human plasma. The cross-reactivity of cIGF-I in a polyclonal hIGF-I radioimmunoassay was 60% of that observed with hIGF-I. The availability of recombinant cIGF-I will facilitate investigations into the role of IGF-I in chicken growth and development.

Amino Acid Sequence↗
PubMed1992

[Laparoscopic appendectomy: 2 cases at a regional hospital].

The surgical technique for removal of the appendix has remained unaltered for many years. Recently, laparoscopic surgery has been introduced with advantages for many patients. We report 2 patients with acute appendicitis who were operated on with laparoscopic surgery at a regional hospital in southern Chile. This technique may become routinely available to general surgeons in the near future.

Adult↗
PubMed1992

Implications of audiologic success.

In the last few years, a clinical consensus has emerged regarding the necessary components of an early management program for hearing-impaired children. Early detection of hearing loss, a parent-infant and supervised nursery program, and optimal amplification provisions are among these components. This paper explores the possible implications for those profoundly hearing-impaired children whose auditory skills become sufficiently developed, as a result of an intensive auditory management program, to permit them to function in a primarily auditory (or auditory/visual) mode under most circumstances. The additional social/vocational/cultural opportunities afforded by a successful auditory habilitation program can require painful choices by many such children. Pursuant to fulfilling their clinical responsibilities regarding amplification, audiologists contribute to this situation.

Audiology↗
PubMed1991

Insulin-like growth factor (IGF)-I and IGF-II binding to an IGF binding protein. An investigation using chemical modification of tyrosine residues as a structural probe for the sites of interaction.

We have investigated insulin-like growth factor (IGF)-I and IGF-II binding to bovine insulin-like growth factor binding protein-2 (bIGFBP-2) using chemical modification to locate sites on the IGF involved in the binding interaction. bIGFBP-2 was incubated with either recombinant human (hIGF-I) or purified ovine (oIGF-II) to form a mixture of bound and free IGF. Sites of interaction between the binding protein and IGF were then probed by iodination of the available tyrosine residues. Subsequently, the mixture of free IGF and IGF.bIGFBP-2 complex was resolved by neutral chromatography, and the IGF component of the complex with bIGFBP-2 was recovered by reverse-phase high performance liquid chromatography at pH 2.1. The tyrosine labeling patterns of the two populations of IGF, one iodinated while free and the other iodinated while associated with binding protein, were determined following endoproteinase Glu-C peptide mapping. Binding of hIGF-I or oIGF-II to bIGFBP-2 resulted in reduced iodination of the tyrosines in both hIGF-I and oIGF-II that are near the carboxyl-terminal, Tyr-60 and Tyr-59, respectively. The reduction in labeling of these tyrosine residues was 2-fold and 6-fold for hIGF-I and oIGF-II, respectively. On the other hand, labeling of the other 2 tyrosines in hIGF-I and oIGF-II was not different between the free and complexed growth factors. From these results we conclude that Tyr-60 and Tyr-59 in the carboxyl-terminal regions of hIGF-I and oIGF-II, respectively, are either directly involved in the binding reaction or lie in a region of the IGF molecule encompassed by the association with bIGFBP-2. Conversely, the labeling pattern of the other tyrosines, Tyr-24 and Tyr-31 in hIGF-I and Tyr-2 and Tyr-27 in oIGF-II, implies that they are not involved in binding to bIGFBP-2. To examine the role of IGF tyrosine residues in the association with bIGFBP-2, we prepared nonradioactive 127I-labeled oIGF-II. In bIGFBP-2 competition binding assays, 127I-labeled oIGF-II was 2.5-fold and 5-fold less potent than native oIGF-II when competing for binding of 125I-labeled IGF-I or IGF-II tracers, respectively. We interpret these results as indicating that at least 1 tyrosine in oIGF-II is involved in binding to bIGFBP-2.

Animals↗
PubMed1991

A case-control comparison of durability and cost between implanted reservoir and percutaneous catheters in cancer patients.

A case-control study was performed to compare the durability and cost of implanted reservoir catheter systems with percutaneous central venous catheters. Twenty cancer patients had reservoir systems placed in 1985 for chemotherapy delivery. The control group consisted of 60 cancer patients, matched according to age, sex, and diagnosis who were part of a group of more than 700 patients with percutaneous catheters inserted during the same period. The reservoir catheters were found to function for a significantly (P less than 0.0001) longer time (495 +/- 54 days) compared to the percutaneous catheters (197 +/- 22 days). The total cost for each system was calculated by adding the charges for an average insertion (reservoir = $1738, percutaneous = $562) to the maintenance charges accumulated over the catheters' lifespan. Reservoir catheters were associated with a significantly greater total cost than percutaneous catheters ($2233 +/- 54, $1453 +/- 102, respectively) but, if the total cost was spread out over the lifespan of the catheter by dividing the total cost by duration of use, reservoir catheters can be less expensive on a per diem basis. The break point occurs at approximately 6 months. For use less than 6 months, percutaneous catheters are cheaper primarily because of their lower insertion costs, but, for longer periods, reservoir catheters become cheaper because of lower maintenance costs and because a second percutaneous catheterization would likely be necessary.

Adult↗
PubMed1991

Breast cancer and second primary ovarian cancer in dermatomyositis.

We report six female patients with breast cancer who developed dermatomyositis and compare our data with those from other reports. The development of dermatomyositis in two patients led to the discovery of a second primary ovarian carcinoma, whereas the development of dermatomyositis in another two patients led to the discovery of recurrent breast cancer. In three patients the diagnosis of dermatomyositis preceded the diagnosis of breast cancer, while the rest developed dermatomyositis after the diagnosis of breast cancer. A parallel clinical course of dermatomyositis and breast cancer was seen in only one patient. Coexisting dermatomyositis and breast cancer is a rare phenomenon, and dermatomyositis that develops during the course of breast cancer may indicate the occurrence of a second primary malignancy or recurrent breast cancer. The onset of dermatomyositis may precede, coincide with, or follow the diagnosis of breast cancer. The clinical course of dermatomyositis sometimes, but not always, parallels the course of breast cancer. There are no specific clinical or laboratory markers to distinguish patients with dermatomyositis who have malignancy from those without cancer.

Adult↗
PubMed1991

A rapid method for the cytodiagnosis of multibacillary leprosy.

The diagnosis of leprosy is made clinically with histologic and cytologic confirmation. The traditional methods for making the cytologic diagnosis of leprosy take time and training, making them unsuitable for rapid office use. We describe a simple, rapid method for cytologic confirmation of the diagnosis of multibacillary leprosy using slit skin scrapings.

Adult↗
PubMed1991

Inbreeding in Saguenay-Lac-St-Jean (Quebec, Canada): a study of Catholic Church dispensations 1842-1971.

Saguenay-Lac-St-Jean (SLSJ) is a rather geographically isolated region of Quebec which shows a high occurrence of hereditary disorders. It has been suggested that high inbreeding might explain this situation. We studied the inbreeding in the SLSJ region by 10-year periods from 1842 till 1971 using the Catholic Church dispensations. The values of the mean inbreeding coefficient were found to be low during the whole period, reaching a peak of 22.94 x 10(-4) during the period 1902-1911. The values observed in the SLSJ were lower than those found in most regions of Quebec and similar to those reported in European populations.

Catholicism↗
PubMed1991

4-Hydroperoxycyclophosphamide purging of breast cancer from the mononuclear cell fraction of bone marrow in patients receiving high-dose chemotherapy and autologous marrow support: a phase I trial.

We designed an ex vivo bone marrow treatment for breast cancer patients receiving high-dose chemotherapy and autologous bone marrow support (ABMS), using 4-hydroperoxycyclophosphamide (4-HC), an active derivative of cyclophosphamide with known activity against breast cancer. This phase I bone marrow purging trial used ficoll-separated mononuclear cells (MNC) (devoid of granulocytes and RBCs), as opposed to the buffy coat. Twenty-five patients with metastatic breast cancer were studied. Patients received three cycles of the Adriamycin (doxorubicin; Adria Laboratories, Columbus, OH), fluorouracil, and methotrexate (Duke AFM) regimen, followed by marrow harvest. An MNC fraction of marrow was prepared and treated with 4-HC in concentrations of 20 micrograms/mL (four patients), 40 micrograms/mL (four patients), 60 micrograms/mL (nine patients), or 80 micrograms/mL (eight patients) and cryopreserved. Patients then received high-dose systemic cyclophosphamide, cisplatin, and carmustine, followed by infusion of the purged marrow. The study end point was marrow engraftment, defined as WBC count greater than 1,000 cells per microliter. At the first three dose levels (20, 40, and 60 micrograms/mL 4-HC), there was no significant delay in time to engraftment (19, 20, and 23 days, respectively) compared with the unpurged historical controls (17 days). At 80 micrograms/mL, engraftment was significantly delayed compared with the lower concentrations (P = .027), and further escalation of 4-HC was not attempted. A significant correlation was observed between the time of leukocyte engraftment and the 4-HC concentration (P = .017). With a methylcellulose-based tissue culture assay, we demonstrated a statistically significant correlation between the colony-forming unit-granulocyte-macrophage (CFU-GM) content in the purged marrow and the days to engraftment. Ninety-five percent of patients responded clinically to the entire program, 55% of them completely. Longer follow-up is required to assess the ultimate benefit of intensive therapy on long-term survival.

Adult↗
PubMed1991

The effect of stimulus rate upon common peroneal, posterior tibial, and sural nerve somatosensory evoked potentials.

We examined the effect of stimulus rate on somatosensory evoked potentials (SEPs) following stimulation of the common peroneal nerve (CPN) at the knee, and the posterior tibial nerve (PTN) and sural nerve (SN) at the ankle. We measured the amplitude of P40-N50 and N50-P60 in the PTN-SEP and corresponding amplitude of CPN-SEP and SN-SEP at the rate of 2.3, 3.4, 4.1, and 5.1 Hz. When the stimulation rate was increased from 2.3 to 5.1 Hz, the P40-N50 amplitude decreased by 50% for the CPN-SEP and 20% for the PTN-SEP. Also, the N50-P60 amplitude was reduced by 30% in the CPN-SEP and 20% in the PTN-SEP. In contrast, this change in stimulus rate produced no significant amplitude decline in the SN-SEP. Blocking the peroneal nerve with lidocaine just distal to the stimulating electrodes eliminated the descending peroneal nerve volley and abolished the amplitude attenuation observed with the faster stimulus rate. The findings suggest that at higher rates of stimulation, the afferent volleys induced by the movements that follow mixed nerve stimulation interfere with the SEP produced by electrical activation of the sensory afferents. The interference is greater when the more proximal site of the mixed nerve is stimulated.

Adult↗
PubMed1991

Expression, purification and characterization of secreted recombinant human insulin-like growth factor-I (IGF-I) and the potent variant des(1-3) IGF-I in Chinese hamster ovary cells.

Recombinant human insulin-like growth factor-I (hIGF-I) and a biologically potent variant lacking the N-terminal tripeptide (des(1-3)IGF-I) were produced from transfected Chinese hamster ovary cells. The constructs encoding the signal peptide, sequence of the mature peptide and a C-terminal extension peptide were expressed under the control of a Rous sarcoma virus promoter. Successfully transfected clones secreting correctly processed recombinant hIGF-I or des(1-3)IGF-I were selected by their secretion of IGF-I-like activity into the culture medium. The recombinant peptides were purified to homogeneity as assessed by high-performance liquid chromatography and N-terminal sequence analysis. The purified recombinant peptides exhibited biological potencies equivalent to authentic IGF-I and des(1-3)IGF-I respectively.

Amino Acid Sequence↗