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Biomedical subjects

Y Castonguay

Publications and source records attributed to Y Castonguay.

At least 19 recordsLinked to original sources

Production of a diagnostic monoclonal antibody in perennial alfalfa plants.

The increasing use of monoclonal antibodies (mAbs) in diagnostic reagents necessitates efficient and cost-effective mAb production methods. In blood banks, one of the most routinely used reagents is the anti-human IgG reagent used for the detection of non-agglutinating antibodies. Here we report the production of a functional, purified anti-human IgG, through the expression of its encoding genes in perennial transgenic alfalfa. Transgenic plants expressing the light- and heavy-chain encoding mRNAs were obtained, and plants from crosses were found to express fully assembled C5-1. The purification procedure yielded mainly the H2L2 form with specificity and affinity identical to those of hybridoma-derived C5-1. The ability to accumulate the antibody was maintained both in parental F1 lines during repeated harvesting and in clonal material; the antibody was stable in the drying hay as in extracts made in pure water. Also, plant and hybridoma-derived C5-1 had similar in vivo half-lives in mice. These results indicate that plant C5-1 could be used in a diagnostic reagent as effectively as hybridoma-derived C5-1, and demonstrates the usefulness of perennial systems for the cost-effective, stable, and reliable production of large amounts of mAbs.

Animals↗

An alfalfa rubisco small subunit homologue shares cis-acting elements with the regulatory sequences of the RbcS-3A gene from pea.

A genomic clone of RbcS was isolated from an alfalfa (Medicago sativa L. cv. Apica) genomic library and characterized. Although this clone has structural features similar to a functional gene, the second exon is interrupted by a stop codon and thus is not fully translatable in the plant. Sequence analysis of the 5' and 3' noncoding regions of RbcSK-1A showed a high sequence homology to the flanking sequences of the RbcS-3A gene from pea. The regions of homology contain many important cis-regulatory elements shown to be essential for regulation of the RbcS-3A gene in pea. The promoter of this alfalfa rubisco clone was used in a translational fusion to test its ability to control the expression of the GUS reporter gene in an homologous nuclear background. High levels of GUS enzyme activity were recorded. These strong levels are comparable to some exceptionally high levels produced in other studies following the use of photosynthesis gene promoters in fusions with the GUS reporter gene.

Base Sequence↗

Differential accumulation of two glycine-rich proteins during cold-acclimation alfalfa.

Two mRNAs, MsaCiA and MsaCiB, encoding for proteins harboring glycine-rich motifs, accumulate in alfalfa during cold acclimation. Fusion polypeptides containing the amino acid sequences deduced from these mRNAs were produced in Escherichia coli and used to raise antibodies. Each antibody cross-reacted specifically with soluble polypeptides, MSACIA-32 and MSACIB, respectively. These polypeptides were detectable only in crowns of cold-acclimated plants, even though MsaCiA mRNA accumulated in both crows and leaves during cold acclimation. The analysis of parietal proteins showed that several MSACIA-related proteins, with a molecular mass of 32, 41 and 68 kDa, did accumulate in leaf cell walls and one of 59 kDa crown cell walls. This diversity is most probably due to a tissue-specific maturation of MSACIA. A discrepancy was found between the time-course of accumulation of MSACIB and the one of the corresponding transcript. These results indicate that timing and localization of MSACIA and MSACIB expression are different, and suggest that this differential expression involves both transcriptional and post-transcriptional events. Comparisons made among six cultivars of contrasting freezing tolerance suggest that low tolerance could be explained by failure to accumulate proteins like MSACIA and MSACIB at a sufficient level.

Acclimatization↗

Changes in ABA and gene expression in cold-acclimated sugar maple.

To determine if cold acclimation of sugar maple (Acer saccharum Marsh.) is associated with specific changes in gene expression under natural hardening conditions, we compared bud and root translatable mRNAs of potted maple seedlings after cold acclimation under natural conditions and following spring dehardening. Cold-hardened roots and buds were sampled in January when tissues reached their maximum hardiness. Freezing tolerance, expressed as the lethal temperature for 50% of the tissues (LT(50)), was estimated at -17 degrees C for roots, and at lower than -36 degrees C for buds. Approximately ten transcripts were specifically synthesized in cold-acclimated buds, or were more abundant in cold-acclimated buds than in unhardened buds. Cold hardening was also associated with changes in translation. At least five translation products were more abundant in cold-acclimated buds and roots compared with unhardened tissues. Abscisic acid (ABA) concentration increased approximately tenfold in the xylem sap following winter acclimation, and the maximum concentration was reached just before maximal acclimation. We discuss the potential involvement of ABA in the observed modification of gene expression during cold hardening.

Journal Article↗

Characterization and mutational analysis of nodHPQ genes of Rhizobium sp. strain N33.

We have shown, by sequencing the nodulation gene region of Rhizobium sp. strain N33 previously isolated from the Canadian high arctic, that the nodHPQ genes are located in a 4.8-kb region downstream of nodBCIJ. The open reading frames of nodHPQ are 747, 906, and 1941 nucleotides long, respectively. The strain N33 genome contains one copy of nodH and two copies of nodPQ that are homologous to those genes in Rhizobium meliloti. Tn5 insertions in the nodHPQ genes of strain N33 did not affect the formation of nodules on the two homologous hosts, Astragalus cicer and Onobrychis viciifolia. Since strain N33 contains the nodBCIJHPQ genes and the recently sequenced nodAFEG genes, we looked for similar host range with R. meliloti. Strain N33 and R. meliloti strains A2 and RCR2011 were shown to induce the formation of root nodules on plants of O. viciifolia. However, strain N33, compared with R. meliloti strains, was able to elicit a few, white, empty, root nodules on Medicago sativa. R. meliloti strains, compared with strain N33, were shown to induce only few nodules containing bacteria on A. cicer. Induction of nod genes transcription in strain N33 was shown to be induced by a variety of flavonoid compounds that are different from those inducing nod genes from R. meliloti.

DNA Mutational Analysis↗

A cold-induced gene from Medicago sativa encodes a bimodular protein similar to developmentally regulated proteins.

A new cold-regulated (COR) gene, msa CIC, was isolated by differential screening of a cDNA library from cold-acclimated crowns of alfalfa (Medicago sativa L. cv. Apica). Transcripts of msa CIC were not detectable in unacclimated alfalfa and accumulated to higher levels in cold-acclimated plants of the cold-tolerant cv. Apica than in those of the cold-sensitive cv. CUF-101. The DNA sequence analysis of a full-length cDNA clone revealed that msa CIC encodes for a putative protein (MSACIC) of 166 amino acids with distinct proline-rich and hydrophobic domains. Protein sequence comparisons indicated that MSACIC is similar to a group of bimodular proteins that are developmentally regulated in other plant species.

Adaptation, Biological↗

A new cold-induced alfalfa gene is associated with enhanced hardening at subzero temperature.

When alfalfa (Medicago sativa L. cv Apica) plants grown at room temperature are transferred to 2 degrees C, the temperature at which 50% of the plants fail to survive (LT50) decreases from -6 to -14 degrees C during the first 2 weeks but then increases to -9 degrees C during the subsequent 2 weeks. However, when plants are kept for 2 weeks at 2 degrees C and then transferred to -2 degrees C for another two weeks, the LT50 declines to -16 degrees C. These changes in freezing tolerance are paralleled by changes in transcript levels of cas15 (cold acclimation-specific gene encoding a 14.5-kD protein), a cold-induced gene. Cold-activation of cas15 occurs even when protein synthesis is inhibited by more than 90%, suggesting that cold-initiated events up to and including the accumulation of cas15 transcripts depend on preexisting gene products. cas15 shows little homology to any known gene at the nucleotide or amino acid level. The deduced polypeptide (CAS15) of 14.5 kD contains four repeats of a decapeptide motif and possesses a bipartite sequence domain at the carboxy terminus with homology to the reported nuclear-targeting signal sequences. Although the relative amount of cas15 DNA as a fraction of the total genomic DNA is similar in cultivars with different degrees of freezing tolerance, its organization in the genome is different. The possible role of cas15 in the development of cold-induced freezing tolerance is discussed.

Acclimatization↗

[Magnetic resonance imaging in the diagnosis and follow-up of false aneurysms after Bentall's operation].

The prevalence and natural history of pseudo-aneurysm (PA) following Bentall procedure has not been established. To determine the follow-up of such patients, we report our experience with magnetic resonance imaging (MRI) used since 1988. At first, spin-echo exam, PA was suspected in 19 patients among 27 patients who underwent aortic valve and ascending aortic replacement between 1980 and 1991. Mean PA diameter was < 4 cm in 13 patients (group 1) and > or = 4 cm in six patients (group 2). Two patients of group 1 were lost at follow-up while two others died after the first spin-echo exam, one death being related to PA formation. Repeated MRIs were done in nine patients of group 1 and all patients of group 2 averaging respectively 2.2 +/- 1.1 exams per patient for group 1 and 1.8 +/- 1.1 for group 2. PA regressed in six patients of group 1 and one patient of group 2 thus suggesting thrombosed PA or postoperative haematoma. PA remained stable in two patients of group 1 and one patient of group 2. One patient of group 1 in whom PA increased at follow-up died suddenly. Four patients of group 2 required surgical correction of their PA. One of these patients died of massive hemoptysis 2 months following reintervention. Recently, addition of cine MRI allows visualization of turbulent flow within the PA thus increasing MRI specificity for PA diagnosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Right ventricular rupture. A complication of postoperative mediastinitis.

Mediastinitis-related right ventricular rupture is an unusual but potentially life-threatening complication of cardiac operations. Between January 1981 and December 1990, a total of 10,182 patients underwent heart operations for ischemic, valvular, and congenital heart disease at the Montreal Heart Institute. Forty-eight patients (0.5%) had postoperative mediastinitis necessitating surgical exploration and sternal debridement. The mediastinum was left open for daily irrigation with povidone-iodine and chest reconstruction was postponed. During treatment, seven patients (0.07%) had right ventricular rupture necessitating immediate surgical repair. All had ischemic heart disease before the operation. There were five women and two men, ages ranging from 52 to 65 years (mean 58 +/- 5 years). Surgical repair consisted of autologous patch covered with omentoplasty assisted with cardiopulmonary bypass. Two patients died, one during the operation of massive hemorrhage and the other 10 days after the operation of uncontrolled sepsis. Five patients survived 2 to 29 months (mean 23 +/- 10 months) after right ventricular rupture, with an overall survival of 71%. Obesity was more frequent in the patients with right ventricular rupture and was found to be a significant risk factor (multivariate analysis, p < 0.05, relative risk 3.22). Histologic examination of the right ventricle in the patient who died after a successful repair revealed fatty infiltration of the right ventricular wall. This may have predisposed the patient toward ventricular rupture. In conclusion, right ventricular rupture, an unusual event in heart surgery, is related to open sternal debridement. Favorable outcome of this complication depends on immediate surgical management, autologous repair, and the use of omentoplasty.

Aged↗

Pseudoaneurysm after Bentall repair: magnetic resonance imaging assessment.

The prevalence and natural history of pseudoaneurysms (PA) following Bentall procedure has not been established. To determine the follow-up of PA in patients, the authors report their experience with magnetic resonance imaging (MRI) since 1988. Spin-echo technique suspected PA in 19 of the 27 patients who underwent aortic valve and ascending aortic replacement between 1980 and 1991. Mean PA diameter was less than 4 cm in 13 patients (group 1) and at least 4 cm in six patients (group 2). Two patients in group 1 were lost at follow-up while two others in this group died after first spin-echo examination (one death was related to PA formation). Repeated MRIs were thus possible in nine patients in group 1 and in all patients in group 2, averaging 2.2 +/- 1.1 examinations per patient for group 1 and 1.8 +/- 1.1 for group 2. PA assessed in six group 1 patients and one group 2 patient suggested thrombosed PA or postoperative hematoma. PA remained stable in two group 1 patients and one group 2 patient. One patient from group 1, in whom PA increased at follow-up, died suddenly. Four patients in group 2 required correction of PA, which had increased in diameter. Recently, addition of cine MRI has allowed visualization of turbulent flow within PA, thus increasing MRI specificity for PA diagnosis. In conclusion, MRI constitutes a reliable noninvasive imaging technique for diagnosis and follow-up of PA after Bentall repair.

Aortic Dissection↗

Accelerated coronary atherosclerosis after cardiac transplantation: major threat to long-term survival.

Accelerated coronary atherosclerosis (ACA) has been documented at autopsy and was noted at coronary angiography in seven patients, 11 to 48 months after cardiac transplantation. To delineate the importance of this problem, the risk factors and the therapeutic approaches in 7 patients who had ACA after heart transplantation were compared with those in 28 patients free of ACA at annual coronary angiography. Ischemic cardiomyopathy was the preoperative diagnosis in all but one patient in the ACA group. The age of the transplant recipients, total myocardial ischemic times and arterial blood pressures at follow-up were similar in both groups. Donor age averaged 31 +/- 3 years in the ACA group and 22 +/- 1 years in patients free of ACA. Preoperative cholesterol, triglyceride and high-density lipoprotein levels were lower in ACA-free patients and the low-density lipoprotein level was higher. At the last follow-up visit, serum lipid levels were similar in both groups. The incidence of acute rejection and of infection was slightly, but not significantly, higher in patients with ACA. The actuarial survival of ACA patients 4 years after transplantation was 30% +/- 20% compared with 100% for patients free of ACA (p less than 0.01). Actuarial rates of freedom from ACA and from death due to ACA were 73% +/- 11% and 81% +/- 11% respectively. Stepwise discriminant analysis showed that older donors and higher pretransplant triglyceride levels were independently related to the development of ACA after cardiac transplantation. In conclusion ACA remains an important cause of late death after heart transplantation. Although therapeutic measures are limited, prevention should focus on strict control of serum lipid levels after transplantation.

Adult↗

Lymphocyte subpopulation monitoring in cyclosporine-treated patients following heart transplantation.

Circulating lymphocyte subpopulations were studied in 18 consecutive patients treated with cyclosporine-prednisone immunosuppression during the first month following heart transplantation. Eleven patients showed no evidence of graft rejection. There were eight episodes of acute rejection demonstrated at endomyocardial biopsy in 7 patients. Three patients were treated with corticosteroids, 3 were treated with rabbit antithymocyte globulin (RATG), and 1 died before treatment (early mortality: 5.6%). Using the monoclonal antibody technique, 150 determinations of lymphocyte subpopulations were performed and were correlated with 72 endomyocardial biopsy specimens. Cyclosporine immunosuppression caused a significant (p less than 0.05) decrease in total lymphocyte count (38%) and in the number of OKT3 (52%) and OKT4 cells (55%). During acute rejection, total lymphocytes and OKT3, OKT4, and OKT8 cells all increased significantly, but the T4 to T8 ratio did not change significantly. Treatment of rejection with corticosteroids resulted in a moderate but not significant decrease in all T-cell types, whereas RATG caused a marked but not selective decrease in all T-cell groups. In conclusion, T cells decrease with cyclosporine immunosuppression and with treatment of rejection and increase at onset of rejection, but the T4 to T8 ratio has no predictive value for the diagnosis and severity of rejection, and the sensitivity of the method does not permit its use to assess the degree of immunosuppression with cyclosporine following heart transplantation.

Adolescent↗

[The surgical alternative in the treatment of terminal cardiomyopathies: cardiac transplantation].

Between April 1983 and November 1985, 14 patients have undergone a heart transplant, or 10 men and 4 women, aged from 17 to 55 years. The surgical indication was a cardiomyopathy in terminal evolutive phase, of which the etiology was ischemia in 8 cases, congestion in 5 cases and rheumatism in one case. There was one operative death (7.1%) from acute rejection and a late death secondary to chronic rejection. The actuarial rate of survival at one and 2 years was 81.9%. Post-operative complications, most frequent at an early stage, were arterial hypertension (9 cases), rejection (8 cases) and renal insufficiency (6 cases). During an average post-operative evolution of 13 months, 17 rejection episodes occurred in 10 patients (71.4%) and 5 patients (35.7%) presented with 7 infectious episodes, one of them severe. Cyclosporin's toxicity appeared in the immediate post-operative period by an always reversible increase of the bilirubin and creatinin levels. On a long term, all patients present with arterial hypertension, accompanied by a moderate renal insufficiency (creatinin between 2.2 and 2.4 mg%). Among the survivors, 9 patients (75%) remain completely asymptomatic and have resumed normal physical activities. Cardiac transplantation has therefore become a valid therapeutic alternative, able to greatly improve survival and the functional condition of patients with terminal cardiomyopathy.

Adolescent↗

[Open-heart surgery after 70 years of age. Results in 136 patients].

Between 1975 and 1983, 136 patients aged over 70 underwent open heart surgery: aorto-coronary by-pass in 67 cases, valve replacement in 46 cases, combined valve and coronary surgery in 20 cases, resection of a left ventricular aneurysm in 2 cases and closure of a ruptured septum in 1 case. There were 13 deaths in the short-term (9.6%). Operative mortality for aorto-coronary by-pass was 4.5%, and that of valve replacement 4.4%, whereas for the combined procedures it was 35%. Non-fatal post-operative complications were frequent, occurring in 40% of patients. Life-table analysis showed 77% survival at 5 years. Of the 111 patients who left hospital, only one was lost to follow-up. At an average follow-up of 26 months, there were 84 patients in functional class I, 17 in class II and 9 in class III. Complications related to the presence of a prosthetic valve occurred in only 3 patients, none of whom required reoperation during the follow-up period. In the elderly, open heart surgery can therefore be carried out at relatively low risk, except where combined procedures are concerned. The long term history, together with the degree of functional improvement achieved, justify cardiac surgery even in the elderly.

Aged↗

Postinfarction ventricular septal defect: delayed closure with prolonged mechanical circulatory support.

Three patients who had ventricular septal rupture and cardiogenic shock complicating an acute myocardial infarction were treated by prolonged circulatory assistance using intraaortic balloon pumping. Hemodynamic stabilization was obtained, and surgical repair was delayed for 19 to 27 days after initiation of circulatory support. All three patients survived the operation and were discharged from the hospital 13 to 19 days after operation. Prolonged intraaortic balloon pumping in such patients is safe and often induces hemodynamic stabilization. The patients may then be operated on under more favorable conditions, without risk of multisystem failure, and after fibrosis has developed around the septal rupture.

Aged↗

[Valve replacement in aged patients. Risk and remote results].

A total of 102 patients aged 65 to 78 years underwent valvular replacement between 1975 and 1980. Calcific aortic stenosis was, by far, the commonest lesion (54 cases), followed by mitral stenosis (16 cases), mitral incompetence (14 cases), aortic incompetence (10 cases) and double valve lesions (8 cases). Sixty four patients underwent aortic valve replacement with 7 early deaths (10,9 p. 100). There were 30 mitral valve replacements with 7 deaths (23,3 p. 100) and 8 patients had double valve surgery with 4 deaths (50 p. 100). Aorto-coronary bypass grafting was associated in 25 cases with a 20 p. 100 mortality. Changes in the technique of peroperative myocardial protection have considerably reduced early mortality which has fallen from over 20 p. 100 to 2,9 p. 100 since the use of cardioplegia with local and systemic hypothermia. There was a higher surgical mortality in patients with poor left ventricular function, cardiomegaly and severe symptomatic incapacity. Non-fatal postoperative complications were common (50 p. 100 of survivors). There were 12 late deaths, 75 p. 100 of which were related to cardiovascular causes. The actuarial survival rate was 65,5 +/- 6 p. 100, 5 years after surgery. When operative mortality was excluded, the 5 year survival rate of the operated patients did not differ from that of the general populations of the same age (79,8 p. 100). Only 5 of our patients were lost to follow-up. Of the survivors, 95,5 p. 100 were asymptomatic or improved by at least one functional grade after an average postoperative period of 30 months. The incidence of late haemorrhagic complications in patients on anticoagulants was 11 p. 100.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Risks and benefits of aortocoronary bypass surgery in patients aged 65 years or more.

Between January 1970 and July 1978, 85 patients aged 65 years or more underwent aortocoronary bypass surgery at the Montreal Heart Institute. The mortality during the operation and the first 29 days thereafter was 12% overall, but was only 5% when the myocardium was protected by the use of cold cardioplegic solutions. Of the 75 patients who survived this period 7 (9%) had a perioperative transmural myocardial infarction. Nonfatal noncardiac complications were more common in these patients than in younger patients, but did not lead to permanent deficits. Three patients died after discharge from hospital, two of cardiac causes. Only one patient had a nonfatal myocardial infarction after discharge. The actuarial 5-year survival rate for all the patients was 80%. After a mean follow-up period of 30 months the condition of 94% of the patients was improved by at least one class of the New York Heart Association functional classification, and 68% were asymptomatic. It is concluded that aortocoronary bypass surgery can be performed in selected older patients with a relatively low in-hospital mortality and morbidity. Symptomatic improvement occurs in almost all such patients. Cardiac catheterization and aortocoronary bypass surgery should therefore be performed in selected older patients with severe angina that is refractory to optimum medical therapy.

Aged↗