Search PubMed⌕ Search

Biomedical subjects

P L Devine

Publications and source records attributed to P L Devine.

At least 37 records · Page 2Linked to original sources
PubMed1998

Serum mucin antigen (CASA) as a marker of amiodarone-induced pulmonary toxicity.

Amiodarone is used to treat life-threatening cardiac arrhythmias. Amiodarone-induced pulmonary toxicity (APT) can be difficult to diagnose. APT may result in increased mucus production and mucin expression. Thus, serum mucin-1 was evaluated as a marker for amiodarone-induced pulmonary toxicity. Concentrations of mucin-1 in peripheral blood were determined using cancer-associated serum antigen (CASA) assay in patients taking amiodarone. Eight of ten patients who developed major amiodarone toxicity had high serum CASA levels. Patients with toxicity had a significantly higher mean rank CASA concentration compared with those without major toxicity. CASA shows potential as a marker for amiodarone-induced toxicity, particularly pulmonary toxicity.

Amiodarone↗
PubMed1998

Monoclonal antibody recognizing a core epitope on mucin.

Monoclonal antibody TH1 (IgM) was prepared by immunizing mice with deglycosylated (TFMSA-treated) cystic fibrosis mucin. TH1 reacted strongly with TFMSA treated cystic fibrosis mucin but not with the fully glycosylated mucin, indicating reactivity with a core mucin epitope. TH1 showed no reactivity with ovine mucin (98% of glycans as sialyl-Tn) but reacted strongly with desialylated ovine mucin, indicating the epitope for this mab was the Tn-antigen (O-linked GalNAc). However, TH1 showed no reactivity with Tn-positive red blood cells, and the binding of TH1 was not inhibited by GalNAc at 2.5 mg/ml, illustrating the importance of the peptide sequence to which the GalNAc is attached. TH1 stained the majority of cancers of the colon, lung, stomach, ovary, breast, and cervix, and the cellular distribution of this antigen in normal tissue suggested reactivity with immature mucin. This antibody appears to be a useful reagent for the detection of immature mucin.

Amino Acid Sequence↗
PubMed1997

Evaluation of a commercial enzyme-linked immunosorbent assay for detection of immunoglobulin M antibody in diagnosis of human leptospiral infection.

The PanBio Leptospira immunoglobulin M (IgM) enzyme-linked immunosorbent assay (ELISA) is a commercially available screening test for the diagnosis of acute leptospiral infection. The ability of the test to diagnose early or recent Leptospira interrogans infection was assessed by testing sera with known microagglutination test (MAT) titers to serovars pomona, hardjo, copenhageni, and australis. The IgM ELISA detected all 41 cases of early or recent leptospiral infection (sensitivity, 100%), with a positive ELISA result seen in many cases before MAT antibody titers reached 1:50. Thirty-eight of 41 patients showed seroconversion (fourfold or greater increase in titer by MAT, 2 of 41 patients had a single sample with elevated titer, and 1 patient from whom leptospires were isolated from a blood sample failed to show MAT titers, despite a seroconversion (negative to positive result) in the ELISA. Follow-up sera obtained from 8 of 12 patients (67%) for 3 to 48 months after the acute stage of illness showed persisting IgM antibody. However, the range of levels detected in these samples (maximum ELISA ratio, 2.0) was lower than the range seen when infection was recent. Reactivity in the IgM ELISA was observed for only 1 of 59 serum samples from asymptomatic donors (specificity, 98%) and 16 of 233 serum samples from patients with Ross River virus, brucella, Epstein-Barr virus, cytomegalovirus, mycoplasma, Q-fever, toxoplasma, hepatitis A virus, Treponema pallidum, or Borrelia burgdorferi infection (specificity, 93%), with the majority of these patients showing lower levels of IgM in comparison to those in patients with leptospiral infection. We conclude that this ELISA is sufficiently sensitive for use as an initial screen for leptospiral infections, with subsequent confirmation of positive test results by MAT.

Analysis of Variance↗
PubMed1997

Serum cytokines and tumour markers in patients with non-small cell carcinoma of the lung.

We compared the serum levels of several cytokines with established tumour markers in 24 patients with non-small cell lung cancer (NSCLC) and 31 patients with benign lung disease (BLD). Cytokine levels were measured using in-house double determinant sandwich ELISAs and tumour markers by a variety of established techniques. There was no correlation between serum cytokines and expression of cytokeratins 18 and 19, MUCI and carcinoembryonic antigen. While no significant difference was observed in any of the cytokines between patients with NSCLC and BLD, patients with metastatic tumour had a significantly higher level of serum tumour necrosis factor alpha and interleukin 10 than those with localised disease (P < 0.015 and P < 0.05 respectively). The serum levels of granulocyte macrophage colony stimulating factor and interferon gamma were no different between these groups. These results suggest immunological effects of NSCLC which tends towards impaired cell mediated immunity in patients with metastatic disease.

Biomarkers, Tumor↗
PubMed1996

Respiratory mucins: identification of core proteins and glycoforms.

At least eight mucin apoproteins are expressed by the tracheobronchial epithelium, but it is not known which, if any, of these are major constituents of the respiratory secretions responsible for the formation of the mucus gel. To address this we have isolated mucins from normal, asthmatic and chronic bronchitic secretions. The asthmatic mucin reduced subunits were fractionated into four populations (I-IV) by anion-exchange HPLC. Amino acid and monosaccharide compositional analysis, as well as M(r) and size measurements, indicate that two of these populations (I and II) are glycoforms of the same or related apoprotein(s) and that the other populations contain two different apoproteins. A panel of antibodies and antisera recognizing the variable number tandem repeat (VNTR) of specific mucin apoproteins did not, as predicted, react with the glycosylated molecules, but after deglycosylation the majority of these probes (with the exception of those to MUC2, which were negative) reacted at a low level with each of the subunit populations. In contrast, an antiserum against a non-VNTR sequence of MUC5AC identified one of the populations (III) as the MUC5AC mucin. The MUC5AC reduced subunit had an M(r) of 2.2 x 10(6) and an RG (radius of gyration) of 57 nm. The genetic identities of the major mucin (populations I and II) and a minor component (population IV) were not established. The MUC5AC mucin was also identified as a major component in the pooled normal secretions from 20 individuals, whereas in a chronic bronchitic sample it was only a minor constituent. Furthermore, in all these different respiratory secretions the MUC5AC mucin appears as a similar biochemical entity, as assessed by Mono Q chromatography and agarose electrophoresis, suggesting that it may have a well-defined pattern of glycosylation in the respiratory tract.

Amino Acid Sequence↗
PubMed1996

Evaluation of four commercial enzyme immunoassays for detection of immunoglobulin M antibodies to human parvovirus B19.

Four commercial enzyme immunoassays (EIAs) for the detection of parvovirus B19-specific immunoglobulin M (IgM) antibodies [Biotrin Parvovirus B19 IgM (Biotrin International, Ireland); Parvoscan B19 IgM (Euro-Diagnostica, Sweden); Parvovirus IgM (Immunobiological Laboratories [IBL], Germany); and human parvovirus B19 IgM (Hillcrest Biologicals, USA)] were compared to indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR). Using IFA as the reference test, high sensitivities (> or = 97%) were observed with all four EIAs, though the specificities of the Biotrin and IBL EIAs (99% and 96% respectively) were significantly higher than those of the Hillcrest and Euro-Diagnostica EIAs (81% and 79% respectively).

Antibodies, Anti-Idiotypic↗
PubMed1996

Evaluation of a commercial enzyme-linked immunosorbent assay for detection of serum immunoglobulin G response to human herpesvirus 6.

A rapid (60-min) commercially available enzyme-linked immunosorbent assay (ELISA) for the detection of immunoglobulin G (IgG) class antibodies to human herpesvirus 6 (HHV-6) was evaluated. The specificity of the ELISA for HHV-6 was confirmed by absorption studies, with the reactivities of HHV-6-positive sera being unaffected by other herpesviruses (cytomegalovirus, herpes simplex virus, and varicella-zoster virus) or the HSB2 cell line used to culture HHV-6. HHV-6 IgG antibody levels in a panel of 502 serum samples were determined by ELISA and an indirect immunofluorescence assay (IFA). Results obtained by the two methods were in close agreement, suggesting that the ELISA provides a suitable test method for the determination of HHV-6 IgG antibodies in a routine clinical laboratory. Both tests were positive in 398 cases (79%), and both were negative in 71 cases (14%), with a different result obtained by IFA and ELISA in only 33 cases (7%). Furthermore, absorption of sera with HHV-6 prior to assay revealed that the majority of these results were false positive (n = 8) or false negative (n = 23) in the IFA (true positives or negatives in the ELISA). Subsequently, the ELISA showed a sensitivity of 99.76% and a specificity of 98.75%. HHV-6-specific IgG levels were also determined in paired serum samples collected from 49 donors--14 with exanthem subitum (ES), 15 with ES which was complicated with central nervous system involvement, and 20 undergoing bone marrow transplantation--in whom HHV-6 infection had been demonstrated by virus isolation and/or PCR. All patients with ES or central nervous system complications showed an increase in HHV-6-specific IgG, indicating that this ELISA may be a useful aid in the diagnosis of these conditions. Furthermore, 14 of 20 patients undergoing bone marrow transplantation showed an increase in HHV-6-specific IgG levels, possibly reflecting a reactivation of HHV-6 in these patients.

Adolescent↗
PubMed1996

Early steps in the biosynthesis of MUC2 epithelial mucin in colon cancer cells.

Expression of the MUC2 mucin has been demonstrated in normal gastrointestinal and respiratory epithelium and in carcinomas of the gastrointestinal and respiratory tracts, breast, ovary, and bladder using RNA probes and (or) monoclonal antibodies reactive with peptide epitopes on the 23 amino acid tandem repeat. Mouse monoclonal antibodies 4F1 and 3A2 were previously obtained by immunization with mucin derived from the LS174T colon cancer cell line and a KLH conjugate of a synthetic MUC2 VNTR peptide. These antibodies react with distinct epitopes on synthetic VNTR peptides and with normal and malignant epithelial tissues. In the present study, we examined the biosynthesis of MUC2 in LS174T colon cancer cells, using these antibodies to immunoprecipitate labelled mucin. A very high molecular mass protein was immunoprecipitated following 1 min pulse labelling with [3H]threonine and [3H]proline. A slight increase in molecular mass was observed over the next 16 min; however, unlike the MUC1 mucin, there was no large difference in apparent molecular mass between the MUC2 protein precursor and fully processed mucin using separation by SDS-PAGE. O-Glycosylation began within 1 h of synthesis of the protein core. Mucin secretion into the culture medium was detected in the 2nd hour following synthesis and was largely completed within 4 h of synthesis. Secreted mucin was far less reactive with these monoclonal antibodies than the precursor protein.

Adenocarcinoma↗
PubMed1995

Methods for separation and deglycosylation of mucin subunits.

Mucins (density 1.35-1.50 g/ml) prepared from human cervical, gastric, respiratory, and salivary secretions were reduced and alkylated to generate reduced mucin subunits. These subunits were separated into different populations analytically by using agarose gel electrophoresis and preparatively by using anion-exchange chromatography. Both techniques separate the molecules primarily on the basis of their inherent charge. A procedure has been developed for the efficient deglycosylation of small amounts of these reduced mucin subunits immobilized on polyvinylidene difluoride. The combination of these procedures has allowed the isolation and identification of glycoforms of a specific mucin gene product in a respiratory mucous secretion.

Blotting, Western↗
PubMed1995

CA15-3, CASA, MSA, and TPS as diagnostic serum markers in breast cancer.

This is the first comparison of the three mucin based tests CA15-3, CASA, and MSA, and the cytokeratin-related TPS assay in breast cancer. The mucin markers were superior to TPS in receiver-operator analysis, though no marker was of use in the diagnosis of malignancy due to low sensitivity. Using cutpoints that gave 95% specificity in benign disease (n = 83), corresponding sensitivities in pre-treatment breast cancer (n = 123: 13 in situ, 54 stage I, 45 stage II, 4 stage III, 7 stage IV) were 17% (CA15-3), 16% (CASA), 13% (MSA), and 8% (TPS), with a strong relationship between marker levels and disease stage. These assays did not always detect the same patients, and the use of CA15-3 combined with CASA gave the highest sensitivity (23%), though this was not significantly better than the use of CA15-3 alone. Despite detecting similar antigens, these assays can show markedly different responses in some patients, indicating that one mucin-based test cannot be substituted for another.

Antigens, Neoplasm↗
PubMed1995

MUC1 in secretory phase endometrium: expression in precisely dated biopsies and flushings from normal and recurrent miscarriage patients.

MUC1 is a cell-surface and secretory product of endometrial epithelium. Immunohistochemical studies carried out using two different antibodies to the mucin-type tandem repeat region of MUC1 indicate a cell-surface location in proliferative phase glands, with intracellular deposits accumulating in the early secretory phase. Commencing 3-4 days after the luteinizing hormone (LH) peak and continuing into the late secretory phase, secretory MUC1 appears in gland lumens. Uterine flushings were collected as a function of time after the LH peak and were analysed using a two-site enzyme-linked immunosorbent assay for MUC1. Low but measurable concentrations were observed up to day 7, while on days 7-13 much higher values were obtained. In women suffering from recurrent spontaneous miscarriage, the concentration of MUC1 in flushings was significantly lower than in the controls on day LH + 10. Lower values were observed on days 7 and 13. Reduced epithelial secretory function and a resultant change in uterine fluid composition are features of endometrium from recurrent miscarriage patients.

Abortion, Habitual↗
PubMed1995

Monoclonal antibodies recognising sialyl-Tn: production and application to immunochemistry.

In order to develop reagents that can detect the exposed sialyl-Tn antigen (NeuAc alpha 2,6GalNAc alpha 1-O-Ser/Thr) on tumour-associated mucins, we have prepared monoclonal antibodies (mabs 3C2 and 3D1, both IgM) against ovine submaxillary mucin (OSM; > 98% of glycans as sialyl-Tn). These mabs showed strong reactivity with OSM and bovine submaxillary mucin (BSM; 50% of glycans as sialyl-Tn) but did not react with desialylated OSM or BSM. Sialic acid at 1 mg/ml did not significantly inhibit mab binding to OSM, suggesting that the linkage to GalNAc may be important for mab binding. 3C2 and 3D1 also showed similar reactivity to sialyl-Tn reactive mab B72.3, and detected B72.3 captured OSM in a sandwich ELISA. In Western blotting of mucus from a patient with a mucinous ovarian tumour, the mabs reacted with high molecular weight (> 200 kDa) species. In immunohistochemistry, these mabs showed strong reactivity with most cancers of the colon, lung, and stomach, and also some tumours of the ovary and breast. There was only limited reactivity in normal tissue from these sites. The antibodies should be useful reagents for the detection of the sialyl-Tn antigen in human cancers.

Animals↗
PubMed1994

Markers for metastatic adenocarcinoma in serous effusion specimens.

A variety of stains have been proposed as useful adjuncts to the morphologic diagnosis of serous effusion specimens. This study evaluates the sensitivity and specificity of nine stains for the detection of metastatic adenocarcinoma in effusions. Mucin cytochemistry with the Periodic acid-Schiff reaction after diastase digestion (PASd) and the mucicarmine method, as well as immunochemical staining for five glycoprotein antigens, was undertaken on 153 effusion specimens with histological and/or clinically confirmed diagnoses. Immunochemical stains included three antibodies to CEA (Dako polyclonal, 4E7 and A5B7) and antibodies to HEA (Ber-EP4), TAG 72 (B72.3), CD15 (LeuM1) and Epithelial membrane antigen (EMA) were evaluated. The sensitivity of the stains for adenocarcinoma (102 cases) was: PASd 37%; mucicarmine 24%; polyclonal anti-CEA 69%; 4E7 52%; A5B7 21%; EMA 91%; B72.3 44%; Ber-EP4 32%; LeuM1 24%. With the exception of EMA, none of the stains reacted with any cases of benign mesothelial cells (11 cases), reactive mesothelial cells (34 cases), or with six cases of mesothelioma. However, EMA staining was present in two cases (6%) of reactive mesothelial cells and all cases of mesothelioma. The optimal combination of stains for use in a panel was polyclonal anti-CEA/B72.3/PASd. Combined results from these three stains yielded a sensitivity of 83% for adenocarcinoma with no false positive results. It is concluded that special staining may provide valuable information to assist in the classification of difficult effusion cases.

Adenocarcinoma↗
PubMed1994

Monoclonal antibodies reacting with the mucin-specific lectin from Sambucus sieboldiana (SSA-M).

Monoclonal antibodies (MAbs) were prepared against the mucin-specific lectin from Sambucus sieboldiana (SSA-M). The majority of MAbs reacted with both free SSA-M and SSA-M bound to porcine mucin. However, MAbs SS16, SS18, and SS19 did not react with mucin-bound lectin, suggesting that these MAbs may react at or near the lectin binding site. Some MAbs also showed reactivity with lectins other than SSA-M (e.g. ConA, Pisum sativum, and Saphora japonica lectins) suggesting possible sequence homology between SSA-M and other lectins. These MAbs should be useful reagents for immunoaffinity purification of SSA-M, characterizing SSA-M structure and binding, and metabolic studies on mucins.

Animals↗
PubMed1994

Mucin expression by transitional cell carcinomas of the bladder.

OBJECTIVE: To examine the presence of a membrane-associated and secreted mucin (MUC1) and a secreted gel-forming mucin (MUC2) in normal and malignant urothelium. MATERIALS AND METHODS: Sections were obtained from archival paraffin blocks from 11 patients with nonmalignant urological conditions and 89 patients with transitional cell carcinomas (TCC). Mucin expression was examined by immunohistochemistry using monoclonal antibodies BC2 and 4F1, reactive with epitopes on the protein core of MUC1 and MUC2 respectively. RESULTS: In normal urothelium MUC1 was limited predominantly to the apical membranes of the umbrella cell layer. MUC1 was present in all cases of TCC, and the pattern of expression divided into three categories: luminal membrane staining only, luminal plus cytoplasmic staining of intermediate +/- basal layers, or staining of only isolated cells or cell groups. These staining patterns were significantly associated with both tumour grade and stage (P < 0.001), with cytoplasmic staining more prevalent in higher grade and stage tumours. MUC2 was not detected in normal urothelium, and was present in 40% of cases of TCC, characterized by intense granular cytoplasmic staining. No association between MUC2 expression and either tumour grade or stage was demonstrated. CONCLUSION: MUC1 mucin was expressed by both normal and malignant urothelium, with increased expression characteristic of higher grade and stage tumours. MUC2 expression was found in 40% of tumours but not in normal urothelium. The role of these mucins in the biology of the bladder requires further investigation.

Aged↗
PubMed1994

Screening and monitoring coeliac disease: multicentre trial of a new serum antibody test kit.

A multicentre trial was conducted to evaluate a new test for anti-gliadin antibodies (AGA) in serum (Coeliac Screening Kit, CSK, Medical Innovations Limited, Artarmon, NSW, Australia). The test showed excellent reproducibility for both anti-gliadin IgA and IgG detection. The average intraassay coefficient of variation (CV) was 3.0% for IgA and 2.4% for IgG (n = 6), while the average interassay CV was 6.4% for IgA and 4.3% for IgG (n = 3). By defining a positive test as both IgA and IgG elevated, a sensitivity of 93% in untreated coeliacs (n = 75) was observed. The corresponding specificities in healthy adults (n = 130) and healthy children (n = 77) were > 99% and 100% respectively, while in patients with other gastrointestinal disorders (disease controls) the specificity was 94% (n = 129). The test was also useful in monitoring patients, with anti-gliadin IgA and IgG falling for up to a year after commencing a gluten-free diet (GFD) (12 adults). In some patients however, antibody levels did not reach the normal cutpoint after many months on a GFD, which may reflect the patients' poor adherence to their gluten free diet. The test was superior to the Pharmacia anti-gliadin ELISA, and should be useful as an aid to the diagnosis of coeliac disease, as well as in the follow-up of treated patients.

Adult↗