Pernicious Anemia Patients Research Articles

The cardiovascular and lipid status of pernicious anaemia patients with and without thyroid antibodies.

A study has been undertaken of the cardiovascular and lipid status of patients with pernicious anaemia (PA). There was no evidence of elevated cholesterol, triglyceride and lipoprotein concentrations nor of an increased prevalence of ischaemic heart disease and peripheral vascular disease in those patients who had positive thyroid antibodies compared with those who had not. There were also no significant differences in cardiovascular or lipid status in the antibody positive patients between those who had an exaggerated thyroid stimulating hormone (TSH) response to thyrotrophin releasing hormone (TRH) and those who had a normal or impaired response. It is suggested that an exaggerated response to TRH in these PA patients usually indicates 'compensated euthyroidism' and not 'subclinical hypothyroidism' or 'premyxoedema' and that they are no more at risk for cardiovascular disease than are PA patients with no evidence of thyroid abnormality.

THYROID FUNCTION AND ANTIBODY STUDIES IN PERNICIOUS ANAEMIA

Thyroid antibodies were demonstrated in 57% of thirty pernicious anaemia patients without overt thyroid disease. Elevated basal thyroid stimulating hormone (TSH) levels and an enhaced TSH response to thyrotrophin releasing hormone (TRH) only occurred in thyroid antibody positive subjects; by contrast the thyroid antibody negative subjects in the older age group frequently and undetecable basal TSH levels and an impaired TRH response. Thyroid hormone concentrations provided no absolute evidence of hypothyroidism in any of the patients.

Antibody to parietal cells in patients with duodenal ulcer, and relationship to pernicious anaemia.

Of 160 patients with pernicious anaemia, none had current duodenal ulceration, whereas in a random population of similar age and sex distribution some 5% would be expected to have a duodenal ulcer. Parietal-cell antibody was detected in serum from 8 of 169 men (4-7%) and from 2 to 31 women (6-4%) with duodenal ulceration. None of the 200 duodenal ulcer patients had antibody to intrinsic factor. The prevalence of these antibodies in duodenal ulcer patients was not significantly different from that in control subjects of similar age and sex distribution. The decreased prevalance of duodenal ulcer in pernicious anaemia patients implies that pernicious anaemia must be less prevalent in duodenal ulcer patients than in a random population; but it appears that this cannot be attributed to an absence of gastric autoimmunity in patients with duodenal ulcer. To resolve this disrepancy, we suggest that pernicious anaemia is determined not only by autoimmune reactions, but also by independent genetic and environmental factors which influence the state of the gastric mucosa.

The Mechanism of Hypergastrinemia in Achlorhydria

The effect of protein-rich food, intravenous calcitonin injections, and intragastric instillation of hydrochloric acid on serum gastrin concentrations and gastrin component pattern was studied in hypergastrinemic patients (pernicious anemia) and matched control subjects. Moreover, plasma secretin concentrations were measured during intragastric acidification. The intragastric acidification resulted in rapid fall in serum gastrin concentrations, although not below the upper limit of normal range. The small components, III (gastrin-17-like) and IV (gastrin-13-like), almost disappeared, whereas the concentrations of component I and component II (gastrin-34-like) were less affected. The increase in secretin concentrations after intragastric acidification was smaller in patients with pernicious anemia than in normal subjects, although the difference was not significant. In contrast to the results in normal control subjects, neither food nor calcitonin produced significant variations in serum gastrin concentrations and gastrin component pattern of pernicious anemia patients. The failure of food to stimulate and of calcitonin to inhibit release of gastrin in the majority of pernicious anemia patients might suggest that gastrin secretion in these patients is autonomous. However, considering the recently recognized slow metabolic clearance rate of big gastrins, the effect of intragastric acidification suggests that the mechanism for acid inhibition of the antral gastrin secretion is intact in patients with achlorhydria.

Differential Binding to Sepharose-Con A of B12-Binding Proteins from Human Gastric Juice

Human gastric juice was found to contain at least two vitamin B-12 binding substances. One of the proteins which formed a complex with B12 was found to bind to a column containing Sepharose-ConA. Since the protein which bound to Sepharose-ConA was absent in the gastric juice of pernicious anemia patients it was concluded that this protein was intrinsic factor. The ability of intrinsic factor to bind to Sepharose-ConA offers a potential means by which intrinsic factor could be separated from other B-12 binding proteins in gastric juice. The ConA binding properties of intrinsic factor might be exploited in the development of a diagnostic test for pernicious anemia.

The abnormal lower oesophageal sphincter in pernicious anaemia.

Lower oesophageal sphincter pressure has been studied in pernicious anaemia patients and controls using an infused open-tipped system. Resting sphincter pressure was significantly (p < 0.01) lower in the pernicious anaemia patients. After gastric acidification with 0.1 N HCl pressure fell significantly (p < 0.01) in both groups. Following subsequent alkalinization, lower oesophageal sphincter pressure for controls rose significantly (p < 0.001). For pernicious anaemia patients the pressure after alkali was not greater than resting levels. Graded intravenous doses of pentagastrin in controls resulted in a peak pressure change of 38.5 +/- 4.9 mm Hg after the 0.8 mug/kg dose. For patients the peak pressure change was only 13.0 +/- 5.2 mm Hg and occurred after 0.4 mug/kg. Cholinergic stimulation with edrophonium (10 mg) produced a peak pressure change of 20.6 +/- 2.6 mm Hg in controls but only 3.5 +/- 1.0 mm Hg in pernicious anaemia patients (p < 0.001). In addition, no change in lower oesophageal sphincter pressure occurred in patients after stimulation with subcutaneous betazole (1.5 mg/kg). In conclusion, the lower oesophageal sphincter in pernicious anaemia is characterized both by a low resting pressure and a decreased response to endogenous and exogenous stimuli. These results suggest a primary end organ defect and most likely indicate abnormal smooth muscle function in pernicious anaemia. Resting sphincter pressure levels and edrophonium response in age-matched subjects indicate that these changes are not due to aging alone.

Abnormal fatty acid metabolism in peripheral nerves of patients with pernicious anemia.

Fatty acid synthesis from radiopropionate was evaluated in sural nerve biopsy slices from five normal controls and nine patients with pernicious anemia. The nerves were incubated in [(14)C]propionate, the lipids were extracted, and the fatty acid methyl esters were chromatographed by gas-liquid chromatography. In the normal nerves the radiolabel was found primarily in short chain (C12 and C14) fatty acids. The nerves from pernicious anemia patients showed two fatty acids peaks that were not discernible in the normal nerves, and these fatty acids had retention times intermediate to those of myristic (C14.0) and palmitic (C16.0) acids and palmitoleic (C16.1) and stearic (C18.0) acids, respectively. These two peaks (a C15 and C17 fatty acid) contained the bulk of the radioactivity recovered in the fatty acid fraction after incubation with [(14)C]propionate. Catalytic reduction and rechromatography failed to alter the retention time of these compounds suggesting that they are not unsaturated fatty acids. The nerves from the pernicious anemia patients had a decrease in the mean content of normal fatty acids when compared with the nerves from control patients as well as a decrease in the mean synthesis of normal fatty acids as estimated by isotope incorporation after incubation with [(14)C]propionate or (3)H(2)O. Analysis of myelin isolated from the nerves indicated that the changes at least in part were in that fraction.

59Fe 2+- and hemoglobin- 59Fe absorption in human beings do not require gastric juice or intrinsic factor

The intestinal 59Fe absorption from 10-μmole (=0.56 mg) 59Fe 2+ and hemoglobin- 59Fe as well as from 5.0-mg hemoglobin- 59Fe doses was measured with the 59Fe-absorption whole-body retention test in normal volunteers and in vitamin-B 12-treated pernicious anemia patients with gastric atrophy and absolute intrinsic-factor deficiency. A completely normal 59Fe absorption from ferrous iron and hemoglobin iron at the 0.56-mg iron level was observed in all the patients without gastric juice. At the 5.0-mg iron dose level hemoglobin- 59Fe was even better (2-fold) absorbed in patients with absolute intrinsic-factor deficiency and a purified hog intrinsic-factor concentrate of known potency did not influence the already doubled hemoglobin- 59Fe absorption. Depleted iron stores in prelatent iron deficiency resulted in a 4-fold increase of 59Fe 2+ absorption and a 50% increase of hemoglobin- 59Fe absorption also in patients without gastric juice and without intrinsic factor. Gastric juice or a gastric intrinsic factor are therefore not required for intestinal ferrous iron or hemoglobin-iron absorption in humans.

Effect of human serum B12 binders on uptake of vitamin B12 by isolated perfused rat liver.

57Co B12-labeled TCII and MPPB prepared from normal human donors and pernicious anemia patients, exhibited a higher uptake by perfused rat liver than 57Co B12-labeled TCI. This is consistent with faster plasma clearance of radioactivity following intravenous injection of 57Co B12 bound to TCII and MPPB in normal human subjects, compared to a slower clearance of TCI B12.

Diagnostic value of the serum folate assay.

The diagnostic value of the serum folate assay has been assessed in 90 patients, each of whom had a macrocytic anaemia and a low serum vitamin B(12) level. Twenty-nine (32%) patients were found to have anaemia due primarily to folate deficiency. The cause of the low serum vitamin B(12) levels is uncertain in the 22 (25%) patients with normal or borderline vitamin B(12) absorption. The effect of folic acid therapy was studied in four of these patients, and in each case the serum vitamin B(12) rose slowly to a normal level. The serum folate was low in only four (7.5%) of the 54 patients with pernicious anaemia, and the levels rose to normal on treatment with vitamin B(12) alone. A high serum folate occurred in eight (15%) pernicious anaemia patients. A normal serum folate indicated the diagnosis of pernicious anaemia or megaloblastic anaemia following partial gastrectomy. However, a normal serum folate and a very low vitamin B(12) level was found in two patients with idiopathic steatorrhoea. It is concluded that the serum folate assay is a valuable routine test in patients who have a macrocytic anaemia and low serum vitamin B(12). A low folate level makes the diagnosis of pernicious anaemia unlikely and is a strong indication for full investigation of small intestinal function.

Peptides of normal human gastric juice.

Normal human gastric juice has been examined for the presence of peptides. Five pooled resting samples have been found to contain a decapeptide of low chromatographic mobility. The sequential structure of this peptide was shown by analysis, and by comparison with a synthetic sample, to be Leu‐Ala‐Ala‐Gly‐Lys‐Val‐Glu‐Asp‐Ser‐Asp Two of the gastric juice samples also contained a peptide of slightly higher Rf value in smaller quantity. This second peptide, though not characterized completely, appeared to have the structure of a nonapeptide. The amino acid composition differed from that of the decapeptide in that leucine was absent; alanine was the N‐terminal amino acid. These peptides have not previously been reported to be present in normal human gastric juice. The authors have not observed any of these peptides in a small number of samples from pernicious anaemia patients.

Clinical Use of Antibodies to Intrinsic Factor.

Meeting Abstracts1 May 1970Clinical Use of Antibodies to Intrinsic Factor.Sheldon P. Rothenberg, M.D., F.A.C.P., Rajani Kantha, M.D.Sheldon P. Rothenberg, M.D., F.A.C.P.Search for more papers by this author, Rajani Kantha, M.D.Search for more papers by this authorAuthor, Article, and Disclosure Informationhttps://doi.org/10.7326/0003-4819-72-5-795_4 SectionsAboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinkedInRedditEmail ExcerptAlthough the identification of antibodies (AB) against intrinsic factor in serum of some pernicious anemia (PA) patients has raised questions about pathogenesis of this disease, of more practical clinical value has been the use of such AB as an aid to rapid etiologic diagnosis of megaloblastic anemias. Two types of AB have been defined; one blocks intrinsic factor from binding B12, and a second binds intrinsic factor or intrinsic factor-B12. Identification of AB in serum supports with 99% certainty diagnosis of PA. A newly developed radioassay for AB using low substrate concentrations is more sensitive, rapid, and easily adapted to... This content is PDF only. To continue reading please click on the PDF icon. Author, Article, and Disclosure InformationAffiliations: New York, N.Y. PreviousarticleNextarticle Advertisement FiguresReferencesRelatedDetails Metrics 1 May 1970Volume 72, Issue 5Page: 795-796KeywordsAntibodiesAttentionMegaloblastic anemiaPathogenesisPernicious anemia Issue Published: 1 May 1970 PDF downloadLoading ...

Studies on hog intrinsic factor. I. Purification of hog intrinsic factor and its physicochemical properties.

One gram of raw hog intrinsic factor (IF) was administered to each of ten control subjects with no hematological disorders, three pernicious anemia patients, and three total gastrectomy subjects. A B12-binding substance originating from raw hog IF was detected in the sera and urines of seven control subjects, after the oral administration of hog IF, and the serum level of the binding substance was assumed to be 5.0μg/ml or slightly more in the detected cases. In the remaining three control subjects and all cases with pernicious anemia or with total gastrectomy, the substance was not found in their sera and urines either before or after the oral administration of raw hog IF. Based on the finding that the detected substance was immunologically identified with the purified hog IF, it was concluded that “immunoreactive hog IF” or rather hog IF or its split fragment was absorbed from human intestine. The significance of the negative demonstration of immunoreactive hog IF in pernicious anemia and in total gastrectomy was also discussed.

Studies on Hog Intrinsic Factor

One gram of raw hog intrinsic factor (IF) was administered to each of ten control subjects with no hematological disorders, three pernicious anemia patients, and three total gastrectomy subjects. A B12-binding substance originating from raw hog IF was detected in the sera and urines of seven control subjects, after the oral administration of hog IF, and the serum level of the binding substance was assumed to be 5.0μg/ml or slightly more in the detected cases. In the remaining three control subjects and all cases with pernicious anemia or with total gastrectomy, the substance was not found in their sera and urines either before or after the oral administration of raw hog IF. Based on the finding that the detected substance was immunologically identified with the purified hog IF, it was concluded that “immunoreactive hog IF” or rather hog IF or its split fragment was absorbed from human intestine. The significance of the negative demonstration of immunoreactive hog IF in pernicious anemia and in total gastrectomy was also discussed.

Prevalence of pernicious anemia in patients with macrocytic anemia and low serum B12.

Objective: The current research evaluated the prevalence of pernicious anemia (PA) in patients with macrocytic anemia (high MCV) and low serum B12 in Riyadh. Methods: Blood testing was done in 77 patients (males: 45.5%, females: 54.5%) with macrocytic anemia; 84 patients; (males: 23.8%, females: 76.2%) with low serum B12 and 30 healthy subjects. Complete blood count, differential count, folic acid, vitamin B12, intrinsic factor, gastric parietal cell antibodies and holotranscobalamin II were assessed. Results: A total of five subjects from 161 patients had PA; three of these patients had macrocyticanemia (3.90%) and two patients had low serum B12 (2.38%). Significant differences (p<0.05) in some hematological, immunological, biochemical parameters were found in subjects with macrocytic anemia and low serum B12 compared to controls. Conclusions: Pernicious anemia in patients with macrocytic anemia and low serum B12 was for the selected sample size can be assumed to be uncommon in Riyadh, Saudi Arabia.

Proliferation of Endocrine-Like (Enterochromaffin) Cells in Atrophic Gastric Mucosa

Per oral mucosal biopsies obtained from the body and pyloric antrum of normal stomachs and from the body of atrophic stomachs of pernicious anemia patients were studied by light and electron microscopy. The atrophic mucosae contained numerous endocrine-like (enterochromaffin) cells both in the islets of ectopic intestinal epithelium and in the nonintestinalized atrophic glands. The proliferation of such cells, which metabolize pharmacologically potent amines and peptides, probably plays a significant role in the physiology of these stomachs.

Antibody activity against hog intrinsic factor in pernicious anaemia.

Summary. The sera of 100 adult patients with Addisonian pernicious anaemia not previously exposed to hog intrinsic factor were tested for antibody activity against hog intrinsic factor by the charcoal method: 55 sera containing antibody to human intrinsic factor showed antibody activity against hog intrinsic factor; 45 sera had no demonstrable antibody activity against human or hog intrinsic factor. A high degree of cross reactivity between antibody to human intrinsic factor and hog intrinsic factor was thus demonstrated in vitro.Hog intrinsic factor was equally effective in potentiating vitamin B12 absorption to normal levels in pernicious anaemia patients with and without circulating antibody active against hog intrinsic factor. Antibody to intrinsic factor circulating in the blood was therefore considered to have no effect on hog intrinsic factor in the human gastrointestinal tract. The possibility of a direct effect of circulating antibodies on the production and/or release of endogenous intrinsic factor by the gastric mucosa could not be excluded.

Radioiodination of human intrinsic factor.

Human intrinsic factor (IF) saturated with (60)Co-labeled cyanocobalamin ((60)CoB(12)) was purified and then iodinated with (125)I to yield (125)I-labeled IF-(60)CoB(12) preparations of high specific activity. Sephadex G200 and DEAE-cellulose chromatography of the iodinated IF-(60)CoB(12) complex showed coincidence of the major (125)I and the (60)Co radioactivity peaks. During starch-gel electrophoresis (60)Co radioactivity from noniodinated and iodinated complexes migrated to the same extent while (125)I radioactivity from the iodinated complex migrated slightly further anodally than did the (60)Co radioactivity. After the iodinated complex was mixed with antibody to the IF-B(12) complex (antibody II) the (125)I and (60)Co radioactivity were: (a) precipitated in similar amounts by antiglobulin serum. (b) eluted coincidentally in the 19S region on Sephadex G200, and (c) excluded to the same extent from starch gel during electrophoresis. After equilibrium exchange of IF "blocking" antibody (antibody I) for (60)Co-vitamin B(12) on (125)I-labeled IF. (125)I radioactivity from the IF-antibody I complex: (a) was precipitated by antiglobulin serum, (b) was eluated in the 19S region on Sephadex G200 gel filtration, and (c) migrated slowly towards the anode on starch-gel electrophoresis. Urinary excretion of (60)Co radioactivity in pernicious anemia patients after oral administration of (60)Co-vitamin B(12) bound to freshly prepared (125)I-labeled IF was similar to that obtained with noniodinated intrinsic factor. These results show that iodination of IF-(60)CoB(12) complex does not markedly alter the chromatographic, electrophoretic, antigenic, or absorption-promoting properties of IF.

Studies on human gastric intrinsic factor. I. Purification of intrinsic factor from human gastric juice.

1. Everted sacs of guinea pig intestine were useful to assay the intrinsic factor activity of the purified human gastric intrinsic factor when pernicious anemia patients or totally gastrectomized patients were not available.2. The author's purified gastric intrinsic factor was found quite active and the activity was destroyed by heating and peptic digestion.3. Gastric juice from three patients having histamine-fast anacidity without pernicious anemia were confirmed to possess intrinsic factor (IF) by this in vitro assay method.4. Gastric juice from three patients with non-Addisonian megaloblastic anemia gave greater uptake of 57Co-B12 by the sac than that in controls where the sac was incubated with 57Co-B12 alone, showing the presence of intrinsic factor in the gastric juice.5. The gastric juice from pernicious anemia patients showed a rather suppressive effect on 57Co-B12 uptake by the sac as compared with the uptake in controls. The presence of the substance inhibiting B12 uptake in the gastric juice of pernicious anemia patients was discussed.

Studies on Human Gastric Intrinsic Factor

The purified intrinsic factor (IF) materials obtained from normal human gastric juice by a sequential column chromatography using Sephadex G-100 gel, DEAE-cellulose and Sephadex G-50 gel, were further analysed immunologically for evaluating the purity by immunodiffusion and immunoelectrophoresis techniques combined with autoradiography. The antisera were produced after successive injections of the purified materials with Freund's complete adjuvant. Purification was proved successful by these methods. IF was not detected in the gastric juice from pernicious anemia patients. Other human body fluids and tissues, such as saliva, bile, sera and liver homogenates, were shown to have no IF. Peptic and α-chymotryptic digestions destroyed the B12 binding capacity of human gastric IF, but the effect of tryptic digestion was not marked, when tested by radioimmunodifusion. The significance of this immunological technique was discussed.