Elevation of serum fucosyltransferase activities in malignant diseases--a sensitive tumor marker?]

Fucosyltransferase (FT) is considered to be one of the most important glycosyltransferases responsible for the synthesis of cancer-associated carbohydrate chains such as CA19-9 and SLX. To determine whether FT is a sensitive tumor marker, we measured the enzyme activity of FT in sera from 136 cancer patients, 14 patients with benign diseases and 59 healthy controls, by using PA (pyridylamino)-labeled type II biantennary oligosaccharide derived from human serotransferrin as an acceptor substrate. Serum FT activity was significantly elevated in patients with cancer compared to healthy controls. Analysis of the enzyme products using HPLC and various fucosidases with different specificity revealed that alpha 1----3 FT was responsible for most of the elevation of the enzyme activity in sera from cancer patients. It should be stressed that the alpha 1----3FT derived from cancer patients transferred fucose to terminal lactosamine residues of type II biantennary oligosaccharides already attached to sialic acid. This indicates that the substrate specificity is clearly different from that reported in normal sera and tissues. In addition to alpha 1----3FT, some glycosidases including fucosidase were also elevated in sera from cancer patients.     

Acute phase factors in anemia]

In various anaemias the values of 8 acute phase factors were determined simultaneously before and at the end of treatment: seromucoid, sialic acid, acid alpha 1-glycoprotein, alpha 1-antitrypsin, haptoglobin, ceruloplasmin, transferrin and fibrinogen. In iron-deficiency anaemia without coexistent inflammatory changes in organs the levels of 4 proteins--seromucoid, alpha 1-antitrypsin, ceruloplasmin and transferrin, were consistently raised. In iron-deficiency anemia with concomitant infection 4 proteins also were increased, but in place of alpha 1-antitrypsin the haptoglobin level was raised. In megaloblastic anaemia the ceruloplasmin level was increased, and in haemolytic anaemia one factor--sialic acid--was decreased. At the end of treatment the concentrations of certain proteins were changed depending on their specific role in various forms of anaemia and on various additional factors. In iron-deficiency anaemia without coexistent infection the concentration of seromucoid was decreased, and in this anaemia with coexistent infection alpha 1-antitrypsin, haptoglobin, and fibrinogen levels were raised, in haemolytic anaemia only fibrinogen was increased, and megaloblastic anaemia was associated with raised seromucoid level. The therapeutic result was good in all these anaemias with the exception of iron-deficiency anaemia associated with infection in which it was less propitious.     

Clinical relevance of sialic acids determination in serum and synovial fluid in orthopaedic disorders

Sialic acids, derivatives of neuraminic acid, are present as structural components of mucoprotein mainly in the alpha 1 and alpha 2-globulin regions, and they are known to change in diseases associated with acute inflammation or tissue necrosis. The present study was performed to clarify the significance of measurements of sialic acids in the serum and synovial fluid of patients with diseases in the field of orthopaedic surgery. 1. Serum sialic acids were markedly high in cases of acute pyogenic diseases and showed moderately high values in stage 2 or 3 of rheumatoid arthritis (RA). There was a close correlation with ESR in RA cases and with CRP in cases of acute pyogenic diseases. Differences in the correlation with ESR and CRP were seen in proportion to the severity of the inflammation. 2. Synovial fluid sialic acids were high in cases of RA, and within the normal range in cases of osteoarthritis. The values changed within this range in accordance with treatment. In RA cases, there was a significant correlation with monocytes in the synovial fluid. 3. Serum sialic acids appeared to be sufficiently useful as a parameter of inflammation independent of ESR and CRP, and synovial fluid sialic acids were also considered to be useful for differentiation between RA and OA.     

The acute phase protein response in patients receiving subcutaneous IL-6.

IL-6, tumour necrosis factor-alpha (TNF-alpha) and IL-1 are thought to be the key mediators of the acute phase response although much of the evidence is based on in vitro studies. It is not clear to what extent each of the acute phase proteins are regulated in vivo by each of these cytokines. The aim of this study was to examine the effects of IL-6 treatment in eight patients with cancer on the concentrations of an extensive range of positive and negative acute phase proteins. It was part of a larger investigation to assess the value of IL-6 in the management of chemotherapy-induced thrombocytopenia. IL-6 was administered by a daily subcutaneous injection for 7 days at a dose level of 1, 3, or 10 micrograms/kg/day. Increases in the positive acute phase proteins, serum amyloid A, C-reactive protein, alpha 1-acid glycoprotein, alpha 1-antichymotrypsin, haptoglobin, alpha 1-antitrypsin, fibrinogen, complement component C3, and caeruloplasmin, were observed, with the greatest incremental changes and fastest responses being seen for C-reactive protein and serum amyloid A protein. The negative acute phase proteins transferrin, transthyretin and retinol binding protein all fell to a nadir within 48-96 h after the first IL-6 injection. Increases in complement component C4 were only found in two patients, which may be related to the increase in circulating TNF-alpha concentrations found only in these patients. This study has therefore shown that IL-6 is capable of causing changes in the majority of acute phase proteins in vivo. Although secondary induction of TNF-alpha was not observed in the majority of patients examined, it is still possible however that other cytokines involved in regulation of the acute phase response, such as IL-1, may have been induced and contributed to the overall response.     

Galactosamine-induced alpha 1-antitrypsin deficiency in rats. Alterations in plasma glycoproteins and alpha 1-antitrypsin carbohydrate composition.

Administration of D-galactosamine (GalNH2) is known to produce alterations in plasma glycoprotein levels, including alpha 1-antitrypsin. The authors have studied the effects of GalNH2 on circulating protein bound carbohydrates and on the plasma concentrations of two alpha 1-antiproteases, transferrin, IgG, and albumin in rats. The alpha 1-antiproteases from GalNH2-treated rats were isolated and their molecular weight, isoelectric point, and carbohydrate composition compared with those of control rat alpha 1-antiproteases. Total plasma protein, albumin, and transferrin levels in the GalNH2-treated rats do not differ significantly from those of control rats. Plasma protein-bound carbohydrate is decreased significantly in the experimental animals, compared with controls: sialic acid decreased 60%, neutral sugars decreased 43%, and amino sugars decreased 38%. The concentrations of alpha 1-antitrypsin (AAT) and a higher molecular weight alpha 1-antiprotease designated AP2 are decreased by 79% and 38%, respectively. AAT isolated from the plasma of GalNH2-treated rats contains 2-3 fewer moles of sialic acid, 3 fewer moles of neutral sugar, and 2 fewer moles of amino sugar per mole of antiprotease than AAT isolated from controls. AP2 from GalNH2-treated rats contains 1 fewer mole each of sialic acid, neutral sugar, and amino sugar per mole of antiprotease than AP2 from controls. These alterations are similar to those seen in humans with genetically determined alpha 1-antiprotease deficiency.     

Changes in the acute phase complement component and IL-6 levels in patients with chronic hepatitis C receiving interferon alpha-2b

In order to study the effect of interferon alpha on the levels of acute phase complement proteins in vivo, serum concentrations of C9 and C1-inhibitor (C1-INH) were measured in patients with chronic hepatitis C before and 3 months after the beginning of interferon alpha2b therapy. Serum levels of the activation product of terminal complement pathway, C5b-9, HCV RNA and IL-6 were also determined. IFN alpha treatment significantly (P<0.0001) increased the serum concentrations of both complement proteins. C5b-9 levels were found to significantly decrease during the same period of time. When the patients were divided into responders or non-responders (more or less than 50% decrease in plasma HCV RNA concentrations) C9 and C1-INH levels were elevated only in the responder patients. There was no correlation between the changes of IL-6 levels or the amounts of IFN alpha administrated on one hand, and the changes in the complement protein levels on the other. These findings suggest that the marked increase in the serum concentrations of the acute phase complement proteins is a secondary phenomenon due to the IFN alpha-caused diminution of the viral load and the resulting immune complex-induced complement activation.     

Colorectal goblet cell mucins in familial adenomatous polyposis.

Ninety two tissue blocks from the left colon and 52 from the right colon were obtained from 112 patients with familial adenomatous polyposis (FAP). Tissues from 137 patients with other conditions served as controls. Within the main study a smaller investigation was performed to compare sections from the left and right colon in the same subject. Several well known histochemical techniques were used to investigate possible changes in sulphation, sialic acid structure (loss of O-acetyl substituents), and changes in the ratio of sialic acid to neutral sugars. In patients with FAP, as in controls, there was increased expression of periodic acid Schiff positive mucin and fucose in the right colon. The only difference between patients with FAP and controls was the indirect demonstration of less neutral mucin in the right colon in FAP, but this did not seem to affect neutral sugars binding to UEA-1, PNL, or HPA. As in the general population, a small proportion of patients with FAP showed a lack of O-acetyl substituted sialic acid. Sialic acid heterogeneity probably has a genetic basis, but this is not associated with the genetic defect underlying FAP.     

Elevated serum sialic acid in pregnancy.

OBJECTIVE: Serum sialic acid has recently gained interest as a possible cardiovascular risk factor as well as a potential tumour marker. The effect of pregnancy on serum sialic acid is unclear particularly in the post-partum period. DESIGN: Longitudinal cohort. SETTING: Teaching hospital antenatal clinic. POPULATION: 29 pregnant women, 27 age matched non-pregnant women. METHODS: Specific enzymatic assay for sialic acid. The first serum sample was taken between 37 and 42 weeks of gestation; in 22 women this was followed by a second serum sample at 12 weeks postpartum. Serum sialic acid concentration was also measured in the 27 controls. RESULTS: Mean (SD) serum sialic acid concentration was higher during pregnancy than post-partum: 91.1 (11.1) v 77.5 (11.1) mg/dl (p < 0.001); or in the control group: 66.0 (9.7) mg/dl (p < 0.001). CONCLUSIONS: Serum sialic acid is elevated during pregnancy and postpartum.     

Increased serum concentrations of tumour necrosis factor in beta thalassaemia: effect of bone marrow transplantation.

AIMS: Serum concentrations of tumour necrosis factor-alpha (TNF) were determined in beta thalassemic patients before and after bone marrow transplantation (BMT) to evaluate whether changes in TNF concentrations after BMT were related to immune mediated complications. METHODS: Serum TNF concentrations were determined by enzyme linked immunoassay (EIA) in paired samples from 71 patients with beta thalassemia before and after BMT. Serial samples from 13 patients were also studied for up to six months after BMT. Forty one normal healthy children matched for sex and age were studied as controls. RESULTS: beta thalassemic patients had high serum TNF concentrations before transplantation compared with controls. These were not related to sex, age, duration of disease, number of blood transfusions, transferrin concentrations or splenectomy. DQw1 positive patients showed significantly lower TNF concentrations than non-DQw1 cases. Patients with severe liver fibrosis had significantly higher TNF concentrations. No correlation was found between TNF values and BMT outcome before transplantation but TNF alpha values fell significantly after BMT. The decrease persisted only in patients with successful engraftment. In serial samples studied for up to six months after BMT, TNF values decreased but in four out of five patients with graft rejection and in all five with acute graft versus host disease (GVHD) sharp increases occurred at the time of clinical symptoms. No correlation was found between the degree of GVHD and serum TNF-alpha concentrations nor between TNF-alpha concentrations after BMT and the presence of bacterial, viral, and fungal infections. CONCLUSIONS: About 50% of beta thalassemic patients have increased serum TNF, and the changes after BMT are related to the occurrence of immune mediate complications. The persistence of low TNF concentrations after successful engraftment may be due to the preparative regimen and the lack of adverse immune reactions.     

Binding of Streptococcus mutans SR protein to human monocytes: production of tumor necrosis factor, interleukin 1, and interleukin 6.

To examine the possible implication of protein SR, an I/II-related antigen from Streptococcus mutans OMZ 175 (serotype f), in inflammatory reactions, we tested the immunomodulatory effects of protein SR on human monocytes. Using biotinylated protein, we provide evidence that protein SR binds to human monocytes in dose-, time-, and calcium-dependent manners through specific interactions. These results were confirmed by competition experiments using either soluble human monocyte extract or anti-SR immunoglobulin G. Binding occurred through lectin-like interactions between SR and carbohydrate portions of monocyte membrane glycoproteins, since binding could be inhibited by several sugars, especially fucose and N-acetylneuraminic acid (NANA), which were confirmed by ligand blotting to be the primer ligands recognized by SR on human monocyte extracts. The ability of protein SR to stimulate the production of cytokines by human circulating monocytes was then examined. The release of tumor necrosis factor alpha (TNF-alpha), interleukin 1 beta, and interleukin 6 is time and dose dependent and not affected by the addition of polymyxin B. Activation of monocytes resulted from specific binding of SR to NANA and fucose present on cell surface glycoproteins since TNF-alpha release could be inhibited by sialidase and pronase treatment of monocytes and by NANA and fucose. These results confirm that sialic acid and fucose present on cell surface macromolecules and especially glycoproteins are needed for the binding of SR to monocytes and for the release of TNF-alpha.     

A study of the prognostic role of serum fucose and fucosyl transferase in cancer of the uterine cervix

Serum fucose levels and fucosyl transferase activities have been designated as nonspecific markers of malignancy, and play an important role in the diagnosis of different types of malignancies. In the present study, attempts were made to determine the prognostic significance of these markers in patients with cancer of the uterine cervix after therapy. It was found that both serum fucose and fucosyl transferase, which were elevated in untreated patients declined significantly in patients responsive to therapy at different follow-up intervals, but not in patients unresponsive to therapy.     

Colorectal goblet cell sialomucin heterogeneity: its relation to malignant disease.

Mucus secreted by colorectal cancer differs in three respects from that produced normally: an overall reduction, a loss of O-acetyl substituents in sialic acid, and an increase in neutral mucin. Similar changes have been reported in apparently normal mucosa bordering colorectal cancer. "Normal" left sided colorectal mucosa from 32 patients with rectal cancer was studied. Each case was matched by age and sex to a patient with diverticular disease and a patient with irritable bowel syndrome. Twenty five patients with right sided cancer were matched to patients with Crohn's disease. Sections were stained with mild periodic acid Schiff (mPAS) (selectively stains N-acetyl sialic acid lacking in O-acetyl group) and other closely related techniques. Reactions were graded negative, weak, and intense. An intense reaction was found in 9% of cases; there was no difference between the various matched groups. Phenylhydrazine interposition failed to block the mPAS effect, indicating that a positive result was due to a deficiency of sialic acid with O-acetyl substituents rather than neutral mucin. Different staining patterns in left and right colon were probably due to differing ratios of total sialic acid:fucose. These findings indicate a hitherto unsuspected colorectal goblet cell sialomucin heterogeneity within the general population, but no association with neoplastic disease is apparent.     

Serum free and bound sialic acid and alpha-1-acid glycoprotein in patients with laryngeal cancer

Serum specimens from 35 patients with laryngeal cancer (10 at stage II; 12 at stage III; 1.3 at stage IV) were obtained before therapy was initiated. Concentrations of bound sialic acid (BSA), free sialic acid (FSA), and alpha-1-acid glycoprotein (AAG) were compared to those in serum specimens from 34 healthy controls. The mean levels of serum BSA and AAG were significantly increased in the laryngeal cancer patients versus the controls; there was no significant difference in serum FSA levels between the patients and controls. Mean serum BSA and AAG levels were lowest in the control group and highest in the stage IV patients. As the stage of the cancer advanced, progressively higher levels of serum BSA and AAG were observed. The results indicate that serum BSA and AAG, but not FSA, show correlation with the stage of laryngeal carcinoma.     

Visualization of carbohydrate chains in rabbit salivary glands by means of enzymatic degradation and plant lectins

We investigated the structure of glycoconjugates contained within the secretory end-pieces and ductal segments in the rabbit submandibular and sublingual glands. Glycosidic sequences were examined by means of enzymatic degradation with specific glycosidases (sialidase, alpha-fucosidase, beta-galactosidase, alpha-mannosidase) followed by lectin binding with PNL-HRP, WPL-HRP, WGL-HRP, SBL-HRP, Con A-HRP. It was found that this procedure represents a valid tool for studying carbohydrates, in so far as their characterization and localization were based only on colour reactions. In particular, this research showed that sialic acid was present in the terminal dimers sialic acid-beta-galactose and sialic acid-N-acetyl-D-galactosamine within the submandibular gland, whereas in the sublingual gland it was only present as the sequence sialic acid-beta-galactose. Conversely, fucose had as the subterminal sugar N-acetyl-D-glucosamine in both glands. Also, elucidations about structural sequences concerning other non-terminal sugars were obtained.     

Absence of prolactin gene expression in colorectal cancer.

AIMS: Previous studies documenting hyperprolactinaemia in patients with colorectal cancer have suggested that the tumour is the source of hormone production. The aim of this study was to determine the frequency of hyperprolactinaemia in patients with colorectal cancer before, during, and after surgery, and also to determine whether prolactin is produced by these tumours. METHODS: Serum prolactin concentrations were measured in 20 patients with colorectal cancer before, during, and after surgical resection of their tumours. Samples taken during surgery included peripheral venous blood and blood taken from the main veins draining the tumour. To determine whether the tumour was responsible for the production of prolactin in these patients, paraffin wax embedded sections of tumour specimens were subjected to immunohistochemistry and western blotting using a monoclonal antibody to prolactin. RESULTS: Five patients (three women, two men) had preoperative prolactin concentrations above the normal reference range, although this increase was of clinical importance in only two. After surgical resection of their tumours, prolactin concentrations remained high in both patients. All 20 patients had greatly raised prolactin values at the time of surgery, irrespective of whether this was measured in peripheral blood or in blood taken from veins draining the tumour. All 20 colorectal cancer tissue samples, including those with raised preoperative and/or postoperative prolactin concentrations, were negative for prolactin staining. Frozen tissue was also available in four cases. The absence of prolactin gene expression in these four tumours was confirmed both by repeat immunohistochemistry and by western blotting. A further 50 colorectal cancer cases examined by immunohistochemistry alone were also unreactive for prolactin. CONCLUSIONS: The results of this study suggest that serum prolactin concentrations may occasionally be raised in colorectal cancer patients, but that the tumour is not the source of hormone production.     

Monitoring serum CEA in women with primary breast tumours positive for oestrogen receptor and with spread to lymph nodes.

Serum carcinoembryonic antigen concentrations (serum CEA) in 80 patients with primary breast cancer were measured preoperatively, one month after operation, and thereafter serially every third month. These data were related to histological and morphometric features of the primary breast carcinoma and the lymph node metastases and to clinical follow up data. Analysis of the serum CEA values showed significant correlations with size of tumour, the presence of lymph node metastases, oestrogen receptor, and occurrence of distant metastases. Furthermore, the results indicated that serial determination of serum CEA in the first two years after operation may be useful in monitoring for the occurrence of distant metastases in patients with metastatic spread to lymph nodes and with large (greater than or equal to 2 cm) primary breast tumours positive for oestrogen receptor. In agreement with other studies, however, it was found that the predictive value of serum CEA concentrations in general is weak and costs may prohibit the implementation of the routine assessment of CEA concentrations.     

Absence of prolactin gene expression in colorectal cancer.

AIMS: Previous studies documenting hyperprolactinaemia in patients with colorectal cancer have suggested that the tumour is the source of hormone production. The aim of this study was to determine the frequency of hyperprolactinaemia in patients with colorectal cancer before, during, and after surgery, and also to determine whether prolactin is produced by these tumours. METHODS: Serum prolactin concentrations were measured in 20 patients with colorectal cancer before, during, and after surgical resection of their tumours. Samples taken during surgery included peripheral venous blood and blood taken from the main veins draining the tumour. To determine whether the tumour was responsible for the production of prolactin in these patients, paraffin wax embedded sections of tumour specimens were subjected to immunohistochemistry and western blotting using a monoclonal antibody to prolactin. RESULTS: Five patients (three women, two men) had preoperative prolactin concentrations above the normal reference range, although this increase was of clinical importance in only two. After surgical resection of their tumours, prolactin concentrations remained high in both patients. All 20 patients had greatly raised prolactin values at the time of surgery, irrespective of whether this was measured in peripheral blood or in blood taken from veins draining the tumour. All 20 colorectal cancer tissue samples, including those with raised preoperative and/or postoperative prolactin concentrations, were negative for prolactin staining. Frozen tissue was also available in four cases. The absence of prolactin gene expression in these four tumours was confirmed both by repeat immunohistochemistry and by western blotting. A further 50 colorectal cancer cases examined by immunohistochemistry alone were also unreactive for prolactin. CONCLUSIONS: The results of this study suggest that serum prolactin concentrations may occasionally be raised in colorectal cancer patients, but that the tumour is not the source of hormone production.     

Usual values of haptoglobin, protein-bound sialic acid and blood sedimentation in a population of the Cherbourg region]

The authors studied 3 parameters commonly used for the exploration of an inflammatory syndrome (determination of globular sedimentation rate at the first and second hour, concentration of plasma haptoglobin, sialic acid bound to plasma proteins rate). Haptoglobin and acid sialic assays were solely used in determining a control population (816 patients without any inflammatory reaction). As a matter of fact, the values we found complied with a normal distribution rule and confidence limits with a probability of 95% were obtained.     

Acute phase response in cerebral infarction.

The serum concentrations of the acute phase proteins, C-reactive protein, serum amyloid A protein, and alpha-1-antichymotrypsin were determined in 50 young and middle aged patients with cerebral infarction and in sex and age matched community controls. Of 46 case control pairs, 11 cases but only one control had raised acute phase protein concentrations simultaneously (p less than 0.01); four case control pairs were excluded because of an infectious complication following infarction and an acute phase response. Seven of the 11 patients (64%) with raised acute phase protein concentrations had a history of bacterial infection in the preceding month but of the remaining 35 patients without raised concentrations only four (11%) had such a history (p less than 0.01). In general, the acute phase response was less pronounced and occurred less often than has been reported in patients after acute myocardial infarction. The results suggest that a positive acute phase response is associated with a preceding bacterial infection or with an infectious complication after cerebral infarction. Measurement of acute phase proteins, therefore, could be used to evaluate the possible role of preceding bacterial infection in the development of cerebral infarction and also in the management of these patients as an early indicator of possible infectious complications.     

Developmental expression of glycocomponents in the chick chorioallantoic membrane

Widespread interest has focused on the research of the chorioallantoic membrane (CAM) and its functional contribution to gaseous exchange, calcium reabsorption, water and electrolyte transport during chick embryogenesis. Nevertheless, very little information is available on the glycoconjugate components of this extra-embryonic structure. In the present study, we investigated by lectin histochemistry, the glycosylation pattern expressed in the CAM epithelia during embryonic development. Occurrence of sialic acid-associated glycoproteins was detailed by either specific lectins, which discriminate alpha2,3 and alpha2,6 sialoderivatives, or sialidase digestion combined with appropriate lectins to identify the sialic acid acceptor sugars. Lectin affinities proved to depend greatly on differentiation of the CAM epithelia which showed highest expression of binding sites during the second half of incubation up to hatching. Differences emerged between the chorionic and the allantoic epithelium, regarding qualitative, quantitative and temporal expression of sugar moieties. A cell type-specific distribution of glycocomponents was found in the chorionic epithelium where lectin binding sites were specifically located in the villus cavity cells. In the allantoic epithelium, high and heterogeneous occurrence of sialoglycoconjugates as well as specific presence of fucose residues were evidenced mostly in the granule cells. We conclude from these findings that various glycoconjugates in the CAM could participate in different physiological functions characteristic of the chorionic and the allantoic epithelium.