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Biomedical subjects

G Richer

Publications and source records attributed to G Richer.

At least 19 recordsLinked to original sources
PubMed1996

Detection of a transforming fragment of herpes simplex virus type 2 in clinical specimens by PCR. The Canadian Women's HIV Study Group.

A PCR assay for the sensitive detection of a transforming fragment of herpes simplex virus type 2 (HSV-2) was developed. Oligonucleotide primers were selected in Xho-2, a transforming region of the BglII N fragment of HSV-2. The assay reached a sensitivity endpoint of 10 copies of the Xho-2 subfragment and did not show cross-reactivity with other herpesviruses, including HSV-1. All 42 HSV-2 isolates scored positive in the assay. The Xho-2 PCR assay was evaluated with 216 clinical specimens and the results were compared with those of cell culture. The best protocol for processing specimens contained in viral transport medium included a centrifugation step followed by cell lysis. Of the 107 specimens positive for HSV-2 by culture, 105 were PCR positive (sensitivity, 98.1%). For one of the two falsely negative samples, beta-globin as well as sequences from the HSV-2 DNA polymerase gene could not be amplified. The other sample scored positive in both of these reactions but was indeterminate in duplicate tests by Xho-2 PCR. Two of 109 HSV-2 culture-negative specimens tested positive in the PCR assay (specificity, 98.2%). The latter two samples tested positive in a PCR test for the HSV-2 DNA polymerase gene. This novel tool was shown to be sensitive and specific for HSV-2 sequences and should allow for the investigation of the role of HSV-2 in genital cancers.

Adult↗
PubMed1994

A long-term follow-up study of asymptomatic hepatitis B surface antigen-positive carriers in Montreal.

BACKGROUND/AIMS: Prospective studies from the Far East and Alaska have shown an increased mortality from cirrhosis and/or hepatocellular carcinoma in asymptomatic hepatitis B virus (HBV) carriers. The magnitude of this risk in apparently healthy North American carriers remains undefined. METHODS: The outcomes of 317 asymptomatic hepatitis B surface antigen-positive carriers from the Montreal area were examined after 16 years of follow-up. A majority of carriers were of French Canadian origin, were positive for antibody to hepatitis B e antigen, and had normal serum transaminase levels; institutionalization in orphanages as infants or children was the most important epidemiological risk factor, suggesting horizontal transmission of HBV during childhood. RESULTS: At follow-up, mean age was 46 +/- 8 years; 3 carriers had died of HBV-related cirrhosis, 1 of alcoholic cirrhosis, and 9 of causes unrelated to liver disease. No carrier died of hepatocellular carcinoma; had the risk of hepatocellular carcinoma been similar to that reported from the Far East and Alaska, 17 cases of hepatocellular carcinoma-related deaths would have been expected. During follow-up, the annual negativation rate for hepatitis B surface antigen was 0.7%. CONCLUSIONS: In asymptomatic HBV carriers from Montreal, a majority are "healthy" carriers and remain asymptomatic after 16 years of follow-up and the risk of death from HBV-related cirrhosis and/or hepatocellular carcinoma is low.

Adult↗
PubMed1990

An outbreak of toxoplasmosis in pregnant women in northern Québec.

An ongoing screening program for toxoplasmosis identified a cluster of four women from northern Québec who, over a 4-month period, seroconverted during their pregnancy. An epidemiologic investigation was carried out in an attempt to identify the source of this infection. All potential risk factors were assessed by a questionnaire administered to 22 Inuit women who had delivered babies in the previous year. Seroconversion was significantly associated with skinning of animals for fur (P = .015) and frequent consumption of caribou meat (P = .034). Compared to seronegative women, women who were seropositive were more than four times more likely to have eaten dried seal meat (P = .067), more than six times more likely to have eaten seal liver (P = .064), and more than eight times more likely to have consumed raw caribou meat more than once per week (P = .054). These observations have contributed to the development of guidelines for the prevention of toxoplasmosis in seronegative pregnant women in this arctic region.

Adolescent↗
PubMed1989

Phenotypic heterogeneity studied by immunohistochemistry and aneuploidy in non-small cell lung cancers.

Non-small cell lung cancers (non-SCLC) differ from small cell lung cancers (SCLC) by many clinical features and prognosis. However, recent studies suggest that lung cancer heterogeneity frequently leads to the association of SCLC and non-SCLC in the same tumor. This phenotypic heterogeneity can be analyzed by immunohistochemistry using monoclonal antibodies (Mab) raised against differentiation related antigens. It may have clinical relevance inasmuch as the diversification of malignant cells is a well-known factor of tumor progression and may be due to chromosomal instability because inappropriate gene expression leads to the formation of antigens unrelated to cell lineage. Chromosomal instability in cancer leads to aneuploidy detectable by cell DNA content analysis. In a prospective study, we analyzed, in parallel, the expression of neuroendocrine related antigens by immunohistochemistry and the cell DNA content in frozen specimens from 40 patients who underwent complete surgical resection of primary non-SCLC in an attempt (a) to characterize the phenotypic heterogeneity and (b) to determine whether this heterogeneity is correlated with aneuploidy and clinical staging. Three Mabs were used in association as a marker of neuroendocrine antigen expression (S-L 11.14, MOC-1, and NE-25); reactivity of these Mabs in 9 SCLC and 3 lung carcinoid tissue sections was used as positive control. All SCLC and 2 of 3 lung carcinoids tested were homogeneously positive with Mabs S-L 11.14, MOC-1, and NE-25; 13 of 40 non-SCLC were homogeneously positive and 11 additional specimens focally positive with Mabs S-L 11.14, MOC-1, and NE-25. The frequency of this abnormal phenotype was significantly higher in poorly differentiated squamous cell carcinomas (chi 2 10.08; P less than 0.005), in clinical stage III non-SCLC (chi 2 5.93; P less than 0.02), and in tumors involving mediastinal lymph nodes (chi 2 5; P less than 0.03). The percentage of cells in the modal DNA of G0-G1 phase was significantly lower in non-SCLC homogeneously positive with Mabs S-L 11.14, MOC-1, and NE-25 [27.4 +/- 10.3% (SD)] in comparison with non-SCLC negative with these same Mabs [56.8 +/- 21.3%; P less than 0.01, Mann-Whitney U test]. We conclude that (a) mixed SCLC-non-SCLC differentiation is frequent and can be assessed by immunohistochemistry, (b) neuroendocrine differentiation in non-SCLC is mainly observed in poorly differentiated tumors and in advanced clinical stages, and that (c) this heterotopic phenotype is correlated with aneuploidy and has clinical implications.

Adult↗
PubMed1989

A monoclonal antibody (BL2-10D1) reacting with a bladder-cancer-associated antigen.

A hybridoma cell line secreting an IgM monoclonal antibody (MAb) was produced after immunizing a mouse with RT4 cells and a crude suspension of human bladder carcinoma cells (WHO grades II and III TCC). This MAb reacted with RT4 target cells derived from a human transitional bladder cancer but failed to react with a majority of non-bladder cancer cell lines. Immunohistological studies indicate that this MAb reacts inconstantly with normal bladder: in positive cases only a few superficial cells (5% to 10% umbrella cells) are stained but not intermediate or basal cells of the urothelium. This MAb was evaluated on 118 tumors: it reacted with tumor tissue in a majority of grade I (79.5%) and grade II papillary TCC (77.3%), less with grade III papillary TCC (45%) and very rarely with invasive non-papillary TCC (14%). In cases of flat lesions a strong reactivity of superficial, intermediate and/or basal layer cells was observed in 50% of moderate and severe dysplasia and in all cell layers of carcinomas in situ (CIS)(9/9).

Antibodies, Monoclonal↗
PubMed1989

Antigenic modulation induced by four monoclonal antibodies adsorbed on gold particles (specificity anti-CD4, anti-CD5, anti-CD7, and anti-150-kDa antigen): relationship between modulation and cytotoxic activity of immunotoxins.

The present study concerns the antibody-induced antigenic modulation of CD4, CD5, CD7, and 150-kDa antigens present on cells of the CCRF-CEM human T line. The immunogold electron microscopy method was used, and it was found that the entry routes associated with the various antigen-antibody complexes were different. Thus, the anti-CD7 monoclonal antibody (MoAb) was frequently internalized via the coated structures of the cell membrane, whereas anti-CD5 MoAb was rarely internalized via those structures and anti-CD4 and anti-150-kDa antigens never used this route. The delay required for 50% internalization of the labeled MoAb-receptor complexes was 30 min. 1 h, 2 h, and 9 h for anti-CD7, anti-CD5, anti-CD4, and anti-150-kDa antigen MoAbs, respectively. A shedding of complexes from the cell surface was never observed. The internalized labeled MoAbs were sequentially transferred into endocytic vacuoles, then into fine anastomosed tubulovesicular structures, and then into lysosomes. However, the anti-150-kDa antigen MoAb proceeded directly from endocytic vacuoles to lysosomes. Among the four MoAbs studied, anti-CD7 MoAb was the most abundant in the endosomal compartment (up to 34% of internalized particles) before it proceeded to the lysosomes. The overall valency of the anti-CD7 MoAb-labeled beads (from 3.8 to 14 MoAb molecules per bead) did not modify the intracellular routing. These results suggested that the subcellular fate of MoAbs was an intrinsic property of each MoAb-antigen complex. More importantly, the comparison between the MoAb-induced modulation and the cytotoxic level of the immunotoxin built with the same MoAb suggested that receptor-mediated endocytosis via coated pits, along with an abundant occurrence of the antigen-MoAb complex within the endosomal complex, could correspond to the best set of conditions for the transfer of the toxin moiety of the immunotoxin to the cytosol.

Antibodies, Monoclonal↗
PubMed1989

The protonation of a retinyl Schiff base: a study by FTIR spectroscopy at low temperature in solution.

The hydrogen bonding-protonation equilibrium for retinyl Schiff base/propionic acid or 3-chloropropionic acid systems was examined by Fourier transform infrared spectroscopy in non polar solutions at temperatures ranging from 25 degrees C to about -150 degrees C. The spectra give evidence for the gradual increase in the degree of protonation as temperature is lowered. The bearing of this on applying low temperature spectroscopic results to physiological conditions in rhodopsin research is discussed.

Fourier Analysis↗
PubMed1987

Immunohistochemical distribution of the 52-kDa protein in mammary tumors: a marker associated with cell proliferation rather than with hormone responsiveness.

We have previously described a secreted glycoprotein of mol. wt 52,000 (52-kDa protein) which is induced by estrogen in some human breast cancer cell lines. This protein has been identified as the proenzyme of a lysosomal cathepsin-D-like protease which is secreted in large proportions in breast cancer cells. To determine which information may be generated by this marker when detected in mammary tumors, in comparison with hormone receptors, we used monoclonal antibodies interacting specifically with the 52-kDa protein and its related cellular processed products (mols. wts 48 and 34 kDa). A high concentration of this protein has been shown in proliferative ductal mastopathies and cysts, suggesting its value in detecting high-risk mastopathies. We now present the immunoperoxidase distribution of this protein in breast carcinoma compared to the cytosolic hormone receptors assayed in parallel. In 232 breast cancers, no correlation was found between the cellular 52-kDa protein content and cytosolic estrogen or progesterone receptor concentrations. This absence of correlation was also shown by the constitutive production of this protein by estrogen-receptor-negative breast cancer cell lines and confirmed by double immunostaining of breast cancer cell aspirates showing a dissociation between the cytoplasmic staining of this 52-kDa lysosomal protease and the nuclear staining of the estrogen receptor. These clinical results, associated with the in vitro mitogenic and proteolytic activities of this protein, strongly suggest that the 52-kDa protein staining in tissue is associated with tumor proliferation and/or invasion, rather than with hormone responsiveness.

Antibodies, Monoclonal↗
PubMed1986

Distribution of the Mr 52,000 estrogen-regulated protein in benign breast diseases and other tissues by immunohistochemistry.

A secreted glycoprotein with a molecular weight of 52,000 is induced by estrogen in breast cancer cells and has been purified to prepare monoclonal antibodies. The protein has been detected in some breast cancers but not in normal breast and uterus. In order to study its potential value as a marker, we have tested by immunohistochemistry frozen sections of several normal and malignant tissues and of benign mastopathies. Among different tissues tested, the Mr 52,000 protein was detected only in liver, sweat glands, and some sebaceous glands, and in malignant melanomas and some breast tumors. Other estrogen-responsive tissues (ovary, placenta, endometrium, etc.) gave negative results. Immunoradiometric assay of the Mr 52,000 protein in biological fluid revealed an elevated concentration in cyst fluid (0.5 to 7.4 micrograms/ml), pleural effusions of certain metastatic breast cancer, and sweat. By immunohistochemistry, the Mr 52,000 antigen was also detected in 42% of 129 benign mastopathies. Gynecomastia, fibrous disease, fibroadenoma, and adenosis were mainly negative, whereas ductal hyperplasia and cysts were positive. The Mr 52,000 protein was found mostly in proliferative ducts and in cysts but not in lobular hyperplasia and nonproliferative lesions without cyst. More Mr 52,000 protein was found in postmenopausal patients than in premenopausal patients. We conclude that the Mr 52,000 protein is a marker associated with mammary cysts and proliferative ducts. On the basis of the increased risk of breast cancer in proliferative mastopathies, we suggest that the Mr 52,000 protein is useful for predicting high-risk mastopathies acting as a marker associated with the proliferation of ductal tissue.

Age Factors↗
PubMed1985

Endocytosis of an antibody ricin A-chain conjugate (immuno-A-toxin) adsorbed on colloidal gold. Effects of ammonium chloride and monensin.

An immunotoxin (IT) formed by a specific antibody coupled to the ricin A chain was adsorbed on colloidal gold particles (IT-Au). Binding and internalization of IT-Au in human lymphoblastic CEM cells were studied using electron microscopy. IT-Au showed specific cytotoxic activity toward the target cells. After 1 h at 4 degrees C, IT-Au were linked diffusely to the plasma membrane with 45% of the particles regrouped in clusters. Upon transfer to 37 degrees C, the particles carrying the ligand were regrouped more frequently and internalized into the cell by endocytosis through smooth microinvaginations or coated pits of the plasma membrane. After 15 min, IT-Au was observed in endocytic vacuoles, or receptosomes, in tubular structure near the Golgi apparatus and in lysosomes. Entry of IT-Au into lysosomes was rapid (around 50% of intracellular IT-Au particles after 30 min). NH4Cl or monensin, well-known potentiators of immunotoxin activity, when present in incubation medium, altered neither the processes nor the rate of IT-Au endocytosis. In the presence of either of these substances, IT-Au accumulated in the normal or often enlarged endocytic vacuoles, and entry into the lysosomes was slowed down (50% of particles after 2 h 15 min). We conclude that this intense slowing-down in the speed of IT-Au transportation into lysosomes and the functional modifications of these organelles help to explain the increased efficacy of immunotoxins in the presence of potentiators.

Adsorption↗
PubMed1984

Immunohistochemical detection of the estrogen-regulated 52,000 mol wt protein in primary breast cancers but not in normal breast and uterus.

An estrogen regulated glycoprotein of molecular weight 52,000 is released by metastatic human breast cancer cells in culture. In order to detect this protein directly in human tissues, several high affinity monoclonal antibodies were produced against the 52,000 mol wt protein. Frozen sections of human breast cancer samples were stained by the peroxidase-anti-peroxidase method using these antibodies. In 20 of 25 samples, specific immunoperoxidase staining was observed in the cytoplasm of epithelial cells with six monoclonal antibodies to the 52,000 mol wt protein. The 5 samples that were not stained contained no detectable estrogen receptor. Epithelial cells were not stained in 6 normal mammary glands collected during reduction mammoplasties and in 9 normal uteri, whether tissues were collected during the follicular or luteal phase. The demonstration that the 52,000 mol wt estrogen regulated protein is present in the cytoplasm of some primary breast cancers but absent in normal mammary tissue and uterus indicates its possible use as a tumor marker.

Antibodies, Monoclonal↗
PubMed1983

[Immunotoxins].

Immunotoxins are conjugates between antibodies especially directed against cancer cells and a subunit of a powerful toxin. We used the A-chain of ricin. These conjugates are specifically cytotoxic when used at very low concentrations in vitro and can destroy more than 99.99% of clonogenic cells. The efficacy of immunotoxins was also demonstrated in vivo but is inferior to its in vitro potency. For this reason the first use of immunotoxins in man can be the cleaning up of bone marrow from leukemic cells in the near future.

Animals↗
PubMed1982

Incidence of hepatitis non-A, non-B compared with types A and B in hospital patients.

To establish the relative frequencies of types A, B and non-A, non-B hepatitis, stored samples of blood from all the cases of acute viral hepatitis seen over a period of 9 years in a general hospital for adults were classified according to their type by presently available serologic methods. The study included 456 episodes of hepatitis in 447 patients, distributed as follows: 114 episodes of hepatitis A (25%), 282 of hepatitis B (62%) and 60 of hepatitis non-A, non-B (13%). The episodes of non-A, non-B hepatitis were equally distributed between the sexes, suggesting a mode of transmission different from that of hepatitis A or B, which had male/female ratios of 2.4 and 3.1 respectively. The low proportion of hepatitis non-A, non-B may not reflect its real frequency, since it often escapes clinical recognition.

Adult↗
PubMed1982

[Immunotoxins (author's transl)].

Immunotoxins are hybrid molecules made up of an antibody and of the toxic subunit of a polypeptidic toxin. They act on the principle that the antibody binds the molecule to a cellular antigen which it specifically recognizes, so that the cytotoxic component only kills those cells that bear the antigen. Cell hybridizations obtained by fusion now produce monoclonal antibodies specific enough to bind the antigens present on certain malignant cells. In vitro experiments have demonstrated the high specific activity of immunotoxins against malignant cells, and this seems to be confirmed by in vivo experiments in mice. Such experimental results suggest that immunotoxins may eventually be used in the treatment of cancer as well as for other pharmacological purposes.

Animals↗
PubMed1981

Immune response of the guinea pig to bovine parathyroid hormone antigen: influence of the booster dose on antisera titer and sensitivity.

Thirty guinea pigs were immunized with 40 microgram of bovine parathyroid hormone, bPTH(1-84), and divided into 6 groups with a similar primary response. Each group was boosted twice with a different dose of antigen (0.04 to 40 microgram). The titer (30% binding of tracer) and affinity (% displacement with 320 pg of bPTH(1-84) were studied. The primary response was maximal at 61 days; titers were 1 x 10(-4) or less and affinity was low. The response was maximal 25 days after each booster. After the first, the highest titers, 5.2 +/- 2.6 x 10(-4) (mean +/- S.D.) were seen in the 40 microgram group and a positive correlation was observed between the dose of antigen and the titer (r = .7386, p less than 0.001) for doses greater than 0.6 microgram. This remained true after the second booster although titers were lower in all groups. Affinity was greater after the first booster (35.85 +/- 20.38%, n = 30) than after the second (23.24 +/- 20.73, n = 30, p less than 0.0025), but was similar in all groups. Selected antisera reacted with antigenic determinants in regions 1-34, 53-84 and 35-53 of the bPTH molecule. Cross-reactivity with human PTH(1-84) was maximal in the 53-84 region. In conclusion, antibodies against all regions of bPTH can be raised in the guinea pig. Once a primary response is elicited, best titers are achieved by boosting with high doses of antigen without any detrimental effect on antisera affinity.

Animals↗