The acute phase response in the rodent.

In the rodent, the general response to acute inflammation and tissue damage is characterized by a complex rearrangement in the pattern of concentrations of proteins in the plasma leading to an increase in the sedimentation rate of erythrocytes, an increase in leukocyte concentration in the bloodstream, and a decrease in the hematocrit. Body temperature changes only slightly or not at all. The reasons for the change in plasma concentrations of proteins are changes in their rates of synthesis in the liver. Degradation of plasma proteins is not affected. The details of the acute phase response evolved in the interaction of species with their environment. Therefore, it is not surprising to find differences in the details of the acute phase response among species. For example, alpha 2-macroglobulin is a strongly positive acute phase reactant in the rat, but not in the mouse; C-reactive protein is a strongly positive acute phase protein in the mouse, but is not found in the rat. An inducible acute phase cysteine proteinase inhibitor system, which has evolved from a primordial kininogen gene, has been observed so far only in the rat. The changes in the synthesis rates of acute phase proteins during inflammation are closely reflected by corresponding changes in intracellular mRNA levels. In the liver, the capacity to induce the acute phase pattern of synthesis and secretion of plasma proteins probably develops around birth. Changes in mRNA levels are brought about by changes in transcription rates or by changes in mRNA stability. Kinetics of mRNA changes during the acute phase response differ for individual proteins. The main signal compound for eliciting the acute phase response in liver seems to be interleukin-6/interferon-beta 2/hepatocyte stimulating factor, whereas interleukin-1 leads to typical acute phase changes in mRNA levels only for alpha 1-acid glycoprotein, albumin, and transthyretin. Plasma protein genes are expressed in various extrahepatic tissues, such as the choroid plexus, the yolk sac, the placenta, the seminal vesicles, and other sites. All these tissues are involved in maintaining protein homeostasis in associated extracellular compartments by synthesis and secretion of proteins. Synthesis and secretion of plasma proteins in paracompartmental organs other than the liver is not influenced by the acute phase stimuli.

mRNA for kininogens and thiostatins (major acute phase alpha 1-proteins) in the liver of kininogen-deficient rats.

mRNAs for low and high molecular weight kininogens (1.6 and 3.0 kb in size, respectively) and for two thiostatins (1.6 kb in size) were found in the liver of kininogen-deficient Brown-Norway (BN/Mai Pfd) rats. The levels of mRNAs for thiostatins, but not those for low and high molecular weight kininogens (arising from a single kininogen gene), increased strongly during acute inflammation. The pattern of DNA restriction sites for the kininogen gene and the thiostatin genes in the mutant rat strain was identical to that in at least four other rat strains.

Structure and expression of the genes for major acute phase alpha 1-protein (thiostatin) and kininogen in the rat.

Two major acute phase alpha 1-protein (alpha 1-MAP, also called thiostatin) genes, one kininogen gene and one structurally related pseudogene, were isolated from a Buffalo rat genomic library and characterized. Comparison of the nucleotide sequence in the 5' proximal flanking region (1 kilobase) of a strongly inducible alpha 1-MAP gene with that in the non-inducible rat kininogen gene showed an overall homology of 90%, with a small number of randomly distributed base changes. In the intron downstream of the exon coding for Ile-Ser-bradykinin, the two alpha 1-MAP genes contained sequence sections similar to the section in the human kininogen gene which codes for the carboxyl-terminal chain of high molecular weight kininogen. Various features of the DNA related to gene expression such as initiation and termination sites of transcription, receptor binding sites, a Z-DNA section, and exon/intron splicing sites were identified. mRNAs expressed by alpha 1-MAP and kininogen genes in liver were studied by RNA blot hybridization using specific oligonucleotide probes. The kininogen gene expressed two mRNA species, one coding for high molecular weight kininogen, the other coding for low molecular weight kininogen. The levels of the two kininogen mRNAs in liver did not increase during acute inflammation. Only one type of mRNA, similar in size to low molecular weight kininogen RNA, was expressed by each of the two alpha 1-MAP genes. The levels of both alpha 1-MAP mRNAs increased strongly during acute inflammation.

Gastroesophageal scintigraphy and endoscopy in the diagnosis of esophageal reflux and esophagitis.

The value of gastroesophageal (G/E) scintigraphy in the diagnosis of gastroesophageal reflux was assessed in 51 subjects, who presented with heartburn and had endoscopic evidence of reflux esophagitis. G/E scintigraphy was done using 99mTc sulfur-colloid in acidified orange juice. The G/E reflux index was calculated according to previous reports. The mean (+/- SD) G/E reflux index in 18 patients with severe esophagitis and 30 patients with moderate esophagitis were 1.6% (+/- 1.5) and 3.2% (+/- 5.0), respectively. The mean G/E reflux index in 14 control subjects was 2.4% (+/- 1.1). There was no significant difference between the esophagitis and control groups. Furthermore, if 4% was taken as upper limit of normal, this will include almost all the esophagitis patients and controls. It is concluded that the G/E reflux index based on G/E scintigraphy is of little value in the diagnosis of G/E reflux.

Endoscopic, histological and ultrastructural correlations in the diagnosis of colitis.

The accuracy of the endoscopic diagnosis, found at fiberoptic distal-colonoscopy, was assessed by correlation with the histological findings of multiple colo-rectal biopsies. Of 25 subjects with normal colon at endoscopy, 18 (72%) had normal histology, while 7 (28%) had histological evidence of mild non-specific colitis (6 cases) or Crohn's colitis (one case). Of 15 cases with endoscopic appearance of mild colitis, histological sections showed mild non-specific colitis in 12 (80%), idiopathic ulcerative colitis in one and Crohn's colitis in another. In 12 patients with an endoscopic diagnosis of moderate-severe colitis, histology confirmed moderate to severe idiopathic ulcerative colitis in 9 (75%) and severe Crohn's colitis in 2. Pseudomembranous colitis was confirmed by histology in 2 out of 3 cases detected by endoscopy. Electron-microscopy, although amplifying the histological findings, did not provide any specific diagnostic information. It is concluded that histological studies of multiple colo-rectal biopsies can significantly improve the diagnostic accuracy of distal-colonoscopy. Biopsies should therefore be obtained routinely in colonoscopy even if the appearances look normal.

P3 amylase isoenzyme in patients with cholelithiasis.

The plasma P3 amylase isoenzyme was determined in 25 patients with proven cholelithiasis. In six patients who had a single gallstone and normal pancreas at operation, the total plasma amylase was within normal limits and the P3 amylase isoenzyme was absent. In the remaining 19 patients who had multiple gallstones, pancreatitis was found at surgery in 10. In this group of 10 patients, the total plasma amylase was above the reference range for healthy individuals in only five (50%), while the P3 amylase isoenzyme band was detected in all 10 (100%). It is concluded that P3 amylase isoenzyme is superior to the total plasma amylase in the diagnosis of pancreatitis in patients with gallstone.

In vitro transformation of Syrian hamster epidermal cells by N-methyl-N'-nitro-N-nitrosoguanidine.

The selection of Syrian hamster epidermal cells which do not terminally differentiate has provided a quantitative focus assay for in vitro chemical transformation. One-day-old Syrian hamster epidermal cells plated at 5 x 10(6)/100-mm dish were treated for 5 hr with various concentrations of N-methyl-N'-nitro-N-nitrosoguanidine. After 4 weeks, the normal epidermal cells began to terminally differentiate to keratinized squamous cells and died, but transformed epidermal colonies grew to higher cells densities and appeared as darker areas against a lightly stained normal cell background. Transformed epidermal foci were isolated and subcultured for at least 15 passages, whereas normal epidermal cells could not be subcultured under the same conditions. The transformed cells assumed the typical cobblestone-like morphology of epithelial cells, retained desmosomes and tonofilaments, and were able to use citrulline in place of arginine. Argininosuccinate synthetase (EC 6.3.4.5) activity was significantly higher in the epidermal cells than in fibroblasts. The injection of 5 x 10(6) cells of two transformed epidermal cell lines into athymic nude mice resulted in the formation of tumors which were identified as keratinizing squamous carcinomas.

Gluten enteropathy occurring in chronic hepatitis.

Steatorrhoea and subtotal villus atrophy due to gluten enteropathy was found to occur in a 27 year old female, who had histologically proven chronic active hepatitis six years ago. She had abnormal elevation of immunoglobulins IgG, IgM, and IgA, decreased level of complement C4 and a positive test for HLA-B8. The SGOT was persistently mildly elevated, but her latest liver biopsy (6 years after onset) showed only changes of chronic persistent hepatitis. She had good symptomatic and histological (jejunal biopsy) improvement with gluten-free diet.

In vivo methotrexate transport in murine Lewis lung tumor.

Methotrexate uptake by murine Lewis lung tumor was measured in vivo over a wide dose range. The data were analyzed according to a model previously developed for tissues in which methotrexate uptake is rate limited by transport across the cell membrane. Methotrexate transport in this tumor followed Michaelis-Menten kinetics with a rate constant for permeability (k/K) of 0.012 min-1. The methotrexate binding capacity of dihydrofolate reductase in the tumor was not exceeded at any dose studied. A low membrane permeability in conjunction with a high dihydrofolate reductase level explains the resistance of this tumor to methotrexate.

In vitro inhibitory effects of polymer-linked methotrexate derivatives on tetrahydrofolate dehydrogenase and murine L5178Y cells.

In vitro studies were made on four synthetic polymeric derivatives of the antitumor agent methotrexate (MTX): 1) divinylether-maleic anhydride-MTX (DIVEMA-MTX), 2) poly-L-lysine-MTX (PL-MTX), 3) polyethyleneimine-MTX (PEI-MTX), and 4) carboxymethyl cellulose-MTX (CMC-MTX). They were tested for their ability to inhibit tetrahydrofolate dehydrogenase (dihydrofolate reductase). Their growth inhibition of murine L5178Y leukemia cells was also studied. 1wo of these polymers, DIVEMA-MTX and PEI-MTX, had similar or only slightly reduced activity compared to equivalent concentrations of MTX, whereas PL-MTX and CMC-MTX had significantly higher (1--3 logs) minimal inhibitory concentrations.

Endoscopic, histological and ultrastructural correlations in chronic gastritis.

Thirty-three patients presenting with dyspepsia were examined with the Olympus Fiber-gastroscope. An endoscopic diagnosis was made in each case and multiple gastric biopsies were obtained for histological and ultrastructural assessment. The endoscopic, histological and ultrastructural findings were compared with each other. Of 33 patients endoscoped, 29 were found to have endoscopic evidence of various types of gastritis and all of these latter had histological evidence of some form of gastritis. Histological confirmation of specific types of endoscopically diagnosed gastritis, however, was only found in 3/9 cases of chronic atrophic gastritis (CAG), 10/14 cases of chronic (superficial) gastritis (CG) and in none of six cases of acute gastritis (AG), indicating that endoscopic diagnosis of specific types of gastritis is relatively inaccurate. Endoscopic diagnosis should thus be restricted to presence or absence of gastritis, leaving the specific typing to histological assessment of the gastric biopsies. Multiple gastric biopsies should be obtained even though the gastric mucosa appears normal endoscopically, since histological evidence of gastritis was found in three out of four cases with endoscopically normal gastric mucosa. Comparison of histological diagnosis with electron microscopy showed that generally there is good correlation between the severity of the histological changes and the ultrastructural grade of damage as defined in this study. It would appear that E.M. examination of the gastric biopsies will not significantly increase the diagnostic accuracy of light microscopy, although it has elucidated the various cellular changes which characterize chronic gastritis. The rough surface contours, the large gastric pit and the increasing number of surface microvilli, seen by scanning E.M., aid the grading of the disease process. Some of these changes are reflected in observations made by transmission E.M. where in addition the basal intercellular edema of the mucosal layer and the appearance of electron-dense mucosal lining cells are observed. The latter, which are the counterpart of "intestinal metaplasia", possess mucus granules which resemble those of gastric mucosal lining cells or intestinal goblet cells, or both.

Fulminant disseminated intravascular coagulation in advanced gastric carcinoma.

A case of fulminant disseminated intravascular coagulation (DIC) in a 51-year old man, presenting with bleeding gastric adenocarcinoma, is reported. In spite of initial hematologic improvement by replacement therapy the patient died on the fifth day after admission. The rare association of DIC with adenocarcinoma of the stomach is discussed.

Immunological studies in chronic atrophic gastritis and chronic (superficial) gastritis.

Detection of autoantibodies, HLA typing and immunofluorescence studies on gastric biopsies were carried out in subjects with histologically proven chronic atrophic gastritis (CAG) and chronic superficial gastritis (CG). All were seronegative for parietal cell antibody and did not have pernicious anemia. Except for positive antismooth muscle and antimitochrondrial antibodies in one patient with CAG, autoantibodies (antinuclear, smooth muscle, mitochrondrial, parietal cell) were absent in patients with CAG and CG. Immunofluorescence studies showed that Ig-G and IgA were presented in the lamina propria of all cases with CAG or CG and of subjects with normal gastric histology. Ig-M was seen less often, in about half the cases. Complement C3 was an uncommon finding, being positive in only one case with CAG and one case with CG and in none of the cases with normal gastric histology. Fibrinogen was more commonly seen in patients with CG (5/5 cases) than in those with CAG (3/11 cases). Fibrinogen was found in one case with normal gastric histology. The most consistent fluorescence was obtained with antiparietal cell antiserum. All subjects with CAG showed negative or weak staining only. In contrast, subjects with CG and normal gastric histology had strong specific fluorescence. An increased frequency of HLA-A1 plus HLA-B8 was found in subjects with CAG (20.7% in controls; 40% in CAG).

Relative biochemical aspects of low and high doses of methotrexate in mice.

During low infusion rates of methotrexate (1.0 microng/hr/mouse; plateau plasma concentration, 2 X 10(-8) M), [3H]deoxyuridine incorporation into DNA was inhibited to a significant degree in small intestine and femur marrows. However, incorporation of [3H]thymidine into intestinal DNA was stimulated at this low infusion rate. During high infusion rates of methotrexate (10 microng/hr/mouse, plateau plasma concentration, 4 X 10(-7) M), inhibition of the incorporation of [3H]deoxyuridine at the steady state levels of plasma methotrexate in both the small intestine and femur marrow was significant. In contrast to stimulation at the low infusion rate, incorporation of [3H]thymidine into intestinal DNA at this high infusion rate was inhibited to a significant degree. Inhibition was not statistically significant in femur marrow DNA. The inhibition of [3H]thymidine into intestinal DNA could be reversed by the simultaneous infusion of inosine. Thus, in the in vivo system, an antipurine effect on DNA Synthesis at high methotrexate plasma concentration in the small intestine was observed. This antipurine effect was not apparent at the lower concentrations. The lower concentration, however, could still inhibit [3H]deoxyuridine incorporation into intestinal and femur marrow DNA to a significant enough degree that, if prolonged, it would resultin lethality to the mice. The thymineless state can be maintained for at most 60 hr in mice without lethal toxicity, whereas the antipurine state can be maintained for no longer than 18 hr in mice without some lethal toxicity. These data have important implications in rescue studies using thymidine or leucovorin.

Gastric acid secretion in chronic atrophic gastritis and chronic (superficial) gastritis.

Basal and pentagastrin-stimulated peak acid outputs were determined in 21 subjects with chronic atrophic gastritis and 10 subjects with chronic superficial gastritis. All subjects were Caucasian. The histological diagnosis was based on multiple gastric biopsy specimens obtained through a fibregastroscope. Comparison of the results with those of a previously reported Caucasian control group show that the mean basal acid outputs of subjects with chronic (superficial) gastritis were significantly higher than that of controls and subjects with chronic atrophic gastritis. No significant difference was found in the mean peak acid outputs of controls and subjects with chronic atrophic gastritis or chronic (superficial) gastritis.

Diagnosis of gastric prepyloric and antral lesions: comparison of fiberendoscopy and gastric biopsy with radiology.

Fibergastroscopy and direct-vision gastric biopsy were performed in 47 patients who had a prepyloric or antral gastric lesion on barium meal (single-contrast) examination. Of 27 cases with a radiological diagnosis of prepyloric or antral ulceration, five cases (18.5%) had evidence of ulceration, 12 cases (44.4%) had acute or chronic gastritis and eight cases (29.6%) had normal gastric mucosa, on fibergastroscopy. Multiple gastric biopsy confirmed the presence of acute-on-chronic gastritis (ACG), chronic gastritis (CG) and chronic atrophic gastritis (CAG), with or without intestinal metaplasia (IM) or epithelial atypia (Aty), in 24 cases (89%). Normal gastric mucosa was found in three cases (11%) and malignancy in none. Of 15 cases with a radiological diagnosis of prepyloric or antral malignancy, only three cases (20%) had evidence of adenocarcinoma on endoscopy and biopsy. One case had rounded nodules seen on endoscopy and gastric biopsies showed malignant lymphoma. In two cases with endoscopic suspicion of malignancy, gastric biopsies showed ACG in one and CAG in the other. Gastric biopsies showed histological changes of CG (+/- IM) or CAG (+/- IM) in 11 cases (73%). In five cases with a radiological diagnosis of various prepyloric or antral lesions, endoscopy and biopsy revealed CG (+/- IM) in all and malignancy in none. It is concluded that fiberendoscopy and gastric biopsy are superior to the single-contrast barium meal in the diagnosis of prepyloric or antral gastric lesions. Direct-vision gastric biopsy should be done in all cases since it increased the diagnostic accuracy of fiberendoscopy.

Diagnosis of gastric malignancy by fibreoptic endoscopy and gastric biopsy: Comparison with radiology.

Of 473 patients examined with the fibregastroscope, 27 were found to have evidence of gastric malignant disease on the endoscopic and biopsy results. Of these 27 patients, 20 were initially examined by barium-meal (single-contrast) radiography then by endoscopy and biopsy. The barium-meal study was reported as showing definite malignant disease in two cases (10%), appearances suggestive of malignant disease in 11 cases (55%), and other lesions in eight cases (35%). Endoscopic examination enabled a diagnosis of advanced gastric cancer to be made in 13 cases (65%), of appearances suggestive of malignant disease in two cases (10%), and of other lesions in five cases (25%). Gastric biopsies gave a histological diagnosis of gastric malignant disease in 19 cases (95%), and of appearances suggestive of malignant disease in one case (5%). Furthermore, the histological types of gastric malignant disease were shown by the gastric biopsies. In seven cases without initial screening by barium-meal radiography, endoscopy and biopsy confirmed the diagnosis of gastric cancer in all. It is concluded that endoscopy is superior to radiology in the diagnosis of gastric malignant disease. The study also shows that gastric biopsy further increases the diagnostic ability of endoscopy and should be done in all cases in which gastroscopy is performed.

Effect of 15 (R) 15 methyl prostaglandin E2 on the healing of gastric ulcers: a double-blind endoscopic study.

A double-blind trial was carried out to assess the effect of 15 (R) 15 methyl prostaglandin E2 on the healing of gastric ulcers. The prostaglandin preparation was given orally in doses of 150 mug every 6 hours for two weeks to 10 subjects with proven gastric ulcer. The control group consisted of 10 subjects with proven gastric ulcer, who did not receive prostaglandin treatment. All were in hospital, and at rest in bed. Ulcer healing was assessed endoscopically with a duodenofibrescope. The maximum diameter of the ulcer crater was determined endoscopically just before and after two weeks of treatment. In the prostaglandin group, complete healing was seen in three cases and considerable healing (50% and more) in five cases, and the overall mean healing rate was 63-3%. In the control group, complete healing was seen in no case and considerable healing in one case, and the overall mean healing rate was 17-1%. The difference was highly significant. The results confirm the belief that 15 (R) 15 methyl prostaglandin E2 is effective in promoting the healing of gastric ulcers.20