Strategies to manage hepatitis C virus infection disease burden - volume 3.

The hepatitis C virus (HCV) epidemic was forecasted through 2030 for 15 countries in Europe, the Middle East and Asia, and the relative impact of two scenarios was considered: increased treatment efficacy while holding the annual number of treated patients constant and increased treatment efficacy and an increased annual number of treated patients. Increasing levels of diagnosis and treatment, in combination with improved treatment efficacy, were critical for achieving substantial reductions in disease burden. A 90% reduction in total HCV infections within 15 years is feasible in most countries studied, but it required a coordinated effort to introduce harm reduction programmes to reduce new infections, screening to identify those already infected and treatment with high cure rate therapies. This suggests that increased capacity for screening and treatment will be critical in many countries. Birth cohort screening is a helpful tool for maximizing resources. Among European countries, the majority of patients were born between 1940 and 1985. A wider range of birth cohorts was seen in the Middle East and Asia (between 1925 and 1995).

Immunohistochemical localization of somatostatin receptor SST2A in the rat pancreas.

Somatostatin (SRIF) acts on specific membrane receptors to inhibit exocrine and endocrine pancreatic functions. Five SRIF receptor genes have been cloned, producing six receptor proteins (sst-s). We used a recently developed antibody to localize the sst2A splice variant in the rat pancreas. Western blots identified the sst2A receptor as an 90 kDa glycosylated protein in pancreatic tissue. In tyramide-amplified immunostainings all acinar cells, and the glucagon and pancreatic polypeptide immunoreactive cells (A and PP, respectively) were intensely labeled for sst2A, while no signal was detected in SRIF producing (D) cells. A very few insulin immunoreactive (B) cells were also labeled for sst2A, but the signal in these cells was lower than in exocrine, A or PP cells. Absorption of the sst2A antibody with the receptor peptide abolished specific staining in both immunoblots and tissue sections (negative control). These studies are the first to localize any SRIF receptor subtype in the rat pancreas. The specific localization of sst2A receptor in acinar, A and PP cells if confirmed in humans, would suggest that subtype specific analogs will be useful for the therapeutic regulation of exocrine and/or endocrine pancreatic secretion.

Immunohistochemical localization of somatostatin receptor SST2A in the rat pancreas.

Somatostatin (SRIF) acts on specific membrane receptors to inhibit exocrine and endocrine pancreatic functions. Five SRIF receptor genes have been cloned, producing six receptor proteins (sst-s). We used a recently developed antibody to localize the sst2A splice variant in the rat pancreas. Western blots identified the sst2A receptor as an 90 kDa glycosylated protein in pancreatic tissue. In tyramide-amplified immunostainings all acinar cells, and the glucagon and pancreatic polypeptide immunoreactive cells (A and PP, respectively) were intensely labeled for sst2A, while no signal was detected in SRIF producing (D) cells. A very few insulin immunoreactive (B) cells were also labeled for sst2A, but the signal in these cells was lower than in exocrine, A or PP cells. Absorption of the sst2A antibody with the receptor peptide abolished specific staining in both immunoblots and tissue sections (negative control). These studies are the first to localize any SRIF receptor subtype in the rat pancreas. The specific localization of sst2A receptor in acinar, A and PP cells if confirmed in humans, would suggest that subtype specific analogs will be useful for the therapeutic regulation of exocrine and/or endocrine pancreatic secretion.

Gastrin-producing endocrine cells: a novel source of histamine in the rat stomach.

Gastrin and histamine both potently stimulate secretion of acid into the gastric lumen. How these agents interact and how their release is controlled is poorly understood. Therefore, we decided to look for histamine in the antral portion of the rat stomach where the gastrin-producing G cells are located. We used immunocytochemical methods to visualize histamine, histidine decarboxylase (HDC, the enzyme that converts histidine to histamine), and the type 1 vesicular monoamine transporter (VMAT1, the protein responsible for moving histamine into vesicles for storage and release). We were surprised to find that histamine, HDC, and VMAT1 were all present in G cells. Our results suggest that G cells synthesize and secrete gastrin and histamine. Whether histamine acts in concert with gastrin to stimulate acid secretion, or functions as an autocrine inhibitor of gastrin release remains to be seen.

Cell specific expression of the sst2A and sst5 somatostatin receptors in the rat anterior pituitary.

Somatostatin (SRIF), originally described as a hypothalamic hormone that inhibits the release of growth hormone was subsequently shown to inhibit the secretion of multiple pituitary hormones. Five genes encoding six different SRIF receptors (sst1, 2A, 2B, 3, 4 and 5) have been cloned and mRNAs for all five are expressed in the anterior pituitary. We used double immunostaining to determine which cells in the anterior pituitary bear sst2A and sst5 receptors. Our results show that these two receptors are widely distributed in the pituitary gland and are both present in a large percentage of GH cells. In addition, sst5 occurs in a small population of corticotrophs and a large percentage of lactotrophs whereas sst2A is found in only a few lactotrophs but a large number of corticotrophs. The sst2A receptor is also expressed in about a third of the gonadotrophs and thyrotrophs. Interestingly, sst2A and sst5 receptors colocalize in a small percentage of cells, most likely somatotrophs demonstrating that the same cells can contain multiple sst receptor subtypes. These results indicate that sst subtype specific analogs are likely to be useful for the selective regulation of individual pituitary hormones.

Colocalization of somatostatin receptor sst5 and insulin in rat pancreatic beta-cells.

Somatostatin, also known as somatotropin release-inhibiting factor (SRIF), is secreted by pancreatic delta-cells and inhibits the secretion of both insulin and glucagon. SRIF initiates its actions by binding to a family of six G protein-coupled receptors (sst1, -2A, -2B, -3, -4, and -5) encoded by five genes. Messenger RNA for both sst2 and sst5 have been reported in the rat pancreas, and the sst2A receptor protein has been localized to rat pancreatic alpha and pancreatic polypeptide-secreting cells in the islets as well as to pancreatic acinar cells. In this study we have used double immunostaining to show that the sst5 protein is expressed exclusively in the beta-cells of rat pancreatic islets and localizes with insulin-secreting alpha-cells. The sst5 receptor is not colocalized with sst2A. Thus, in the rat SRIF inhibits pancreatic insulin and glucagon secretion via different sst receptor subtypes.

Substantial production of dopamine in the human gastrointestinal tract.

Considerable urinary excretion of dopamine metabolites indicates that large amounts of dopamine are produced in unknown locations of the body. This study assessed the contribution of mesenteric organs (gastrointestinal tract, spleen, and pancreas) to the total body production of dopamine in humans and examined the presence of the rate-limiting enzyme for dopamine synthesis, tyrosine hydroxylase, in gastrointestinal tissues. Blood sampled from an artery and portal and hepatic veins in eight subjects and from arterial and renal venous sites in other subjects was analyzed for plasma concentrations of dopamine and its metabolites. The activity and distribution of tyrosine hydroxylase was also examined in tissue samples from the stomach and duodenum. Higher concentrations of dopamine and its metabolites in portal venous than arterial plasma indicated substantial production of dopamine by mesenteric organs (12.0 nmol/min) amounting to 42-46% of the renal removal of circulating dopamine metabolites. Tissue samples showed immunoreactive tyrosine hydroxylase in nonneuronal cell bodies and detectable levels of tyrosine hydroxylase in nonneuronal cell bodies and detectable levels of tyrosine hydroxylase enzyme activity. The results show that mesenteric organs produce close to half of the dopamine formed in the body, most of which is unlikely to be derived from sympathetic nerves but may reflect production in a novel nonneuronal dopaminergic system.

Immunohistochemical signal amplification by catalyzed reporter deposition and its application in double immunostaining.

The biotinyl-tyramide substrate of the horseradish peroxidase enzyme has been recently introduced to amplify immunohistochemical signals. We applied either fluorochromeor biotin-conjugated tyramine to improve the detection of different antigens in sections of rat stomach, pancreas, and hypothalamus. A ten- to 100-fold increase in staining efficiency was achieved, depending on the antibody, with either fluorescent or peroxidase detection systems. The amplification method was particularly useful for increasing a weak signal of conventional immunostaining caused by suboptimal tissue fixation. At a very low concentration of the primary antibody, the antigen can no longer be detected by a conventional fluorescent secondary antibody but is still detectable after amplification. When an antibody is used at this very low concentration and is detected by a fluorescent amplification method, another primary antibody, raised in the same host species, can be used and demonstrated with a different fluorochrome in subsequent conventional immunostaining of the same section. In this way it becomes possible to immunostain the same section with two different primary antibodies raised in the same host species. Samples for such double immunostaining are demonstrated here using pairs of monoclonal antibodies (to tyrosine hydroxylase and oxytocin) in the hypothalamus and polyclonal antibodies (to glucagon and neurofilament M) in sections of rat pancreas. Because in many cases the availability of antibodies is limited, the amplification method can be a quick and efficient tool for double immunostaining with antibodies from the same host species.

Diagnostic interpretation of 24-hour intragastric pH-metry using a pattern recognition method.

BACKGROUND: Intragastric pH-metry is a widely used method in the evaluation of gastric acidity, but the interpretation of these data is not standardized. METHODS: The pH-metry data of 60 patients were sorted retrospectively into three groups according to the maximal acid output (MAO) values measured after stimulation by pentagastrin: hypoacid group (MAO < 5 mmol/h, n = 17), normacid group (MAO 10-15 mmol/h, n = 18) and superacid group (MAO > 25 mmol/h, n = 25). Statistical effectiveness of several descriptive statistical values and predefined time-intervals in differentiation between the study groups was analysed by the pattern recognition by independent multicategory analysis (PRIMA) method. RESULTS: The mean pH, the integrated pH and the time-interval of pH > or = 3 values were the most effective parameters for discriminating between each pair of groups. Hypoacid-normacid-superacid: mean pH 6.1 +/- 0.2-4.2 +/- 0.23-1.5 +/- 0.07; integrated pH 8898 +/- 208-5987 +/- 339-2224 +/- 98 pH.min; and pH > or = 3 1440 +/- 0-392 +/- 44-80 +/- 11 min, respectively (+/- SEM). More than a 99.9% separation of the study groups was achieved using these three parameters concomitantly by the PRIMA method, even when comparing 6-h daytime periods of measurements. A relative disagreement was found when reclassifying the hypoacid and normacid patients on the basis of pH records, compared with the primary classification based on the MAO data (11 out of 35 patients, 31%). CONCLUSIONS: The analysis of the mean pH data (instead of medians or [H+]), the AUC-pH and the pH > or = 3 duration, either separately or most beneficially concomitantly, are recommended for the diagnostic interpretation of intragastric pH-metry data. The duration of the diagnostic intragastric pH-metry measurements might be decreased to 6 h by using the PRIMA method.

Vesicular monoamine transporters in the rat stomach.

Cellular distribution of vesicular monoamine transporters (VMATs), known to regulate vesicular storage and release of biogenic amines (i.e., catecholamines, serotonin, histamine, etc.), have been studied in the rat stomach using in situ hybridization histochemistry (ISHH) and immunohistochemical (IHC) techniques. 35S-UTP labeled riboprobes showed that mRNAs of both VMATs are expressed in the gastric mucosa. A combination of ISHH and IHC verified that most of the parietal cells (among other epithelial cells) express mRNA of the peripheral type transporter (VMAT1) while enterochromaffin-like cells (ECL) of the fundic mucosa express mRNA of the central type (VMAT2). In addition, with double fluorescent IHC we detected VMAT1 protein in serotoninergic enterochromaffin cells (EC) of the stomach and in gastrin producing G cells of the antral mucosa. Similarly to the fundus, VMAT2 protein was present in ECL cells and in the enteric plexus. Surprisingly, serotonin- and/or histamine-containing cells in the connective tissue compartments of the stomach (i.e., lamina propria and submucosa), immunoreactive for a mast cell specific antigen, displayed neither VMATI nor VMAT2 immunoreactivity. Distribution of VMATs in the rat stomach support our previous observations on aminergic properties of two important gastrointestinal (GI) epithelial cell populations primarily known for other specific secretory products, i.e. dopaminergic properties of acid producing parietal cells and histaminergic properties of gastrin producing G cells. These data emphasize the existence of a non-neuronal, intrinsic aminergic system in the GI tract.

13C-Urea breath test is superior in sensitivity to detect Helicobacter pylori infection than either antral histology or rapid urease test.

There is no single technique which fulfils the criterion for a reference method to detect Helicobacter pylori (Hp) infection. The aim was to compare the results of antral histology (H), rapid urease test (U) and urea breath test (UBT) from antral biopsy samples in patients having gastric or duodenal lesions during upper GI endoscopy. We used the following methods: 1) biopsy specimens for histology (Warthin-Starry staining); 2) rapid urease test; and 3) 13C-urea breath test with infrared spectrometry. The total number of patients was 166 examined by H, U, and UBT. H, U and UBT were negative (-) in 64 patients and positive (+) in 51. The true positivity and false negativity (%, number of patients in parentheses) of each method based upon the positivity of the other two tests were: H+, U+ (54): UBT+, 94.4% (51) and UBT-, 5.6% (3); H+, UBT+ (57): U+, 89.5% (51) and U-, 10.5% (6); U+, UBT+ (65): H+, 78.5% (51) and H-, 21.5% (14). If Hp infection is considered to be positive when at least two tests detect the presence of Hp, UBT shows the highest sensitivity in comparison to histology of biopsy specimens and urease test. UBT is highly recommended as a screening test for Hp infection in patients presenting upper GI endoscopic alterations.

One year follow-up of patients after successful helicobacter pylori eradication therapy.

UNLABELLED: The aim of this study was to investigate the Helicobacter pylori (Hp) status of patients who underwent successful eradication therapy 1 year prior to the study and to evaluate their current symptoms. METHODS: all of the patients were initially evaluated by oesophago-gastro-bulboscopy and the Hp status was determined by at least two different methods [rapid urease test, histology or urea breath test (UBT)]. The Hp infection was treated with a 1-week triple therapy protocol, and the UBT was repeated 4-6 weeks later. We invited back 110 patients who had negative post-eradication UBT results 12+/-3 months prior to the study period. UBT was repeated and a questionnaire was completed about the previous and present complaints and medication. RESULTS: 80 of the 110 patients (73%) came back for the follow-up. Twenty five patients had peptic ulcer disease, 36 patients had gastritis or duodenitis without erosive lesions, and 19 patients had erosive form of gastritis or duodenitis initially. All of the patients except one in the erosive gastritis group had negative control UBT 1 year after the eradication, which means 1.25% recurrence rate within 1 year. The eradication therapy completely revealed the symptoms of 16 patients in the ulcer group (64%), 13 patients in the gastroduodenitis group (36%, P=0.03 vs. ulcer patients), 10 patients with erosive gastroduodenitis (52%), but this was only temporary. One year after the eradication therapy seven of the ulcer patients (28%), 11 patients with gastroduodenitis (31%) and seven patients with erosive gastroduodenitis (37%) were symptom-free. Most of the patients had epigastric pain (44%), heartburn (43%) and/or abdominal distension (33%). Nine ulcer patients (36%), 10 patients with gastroduodenitis (28%) and five patients with erosive gastroduodenitis (26%) were taking H(2)-blockers regularly. CONCLUSION: the 1-month post-eradication UBT was probable true negative in all of the evaluated cases, since 79 patients (98.75%) were also negative after 1 year. The Hp recurrence rate is very low (1.25%) in a 1-year period. The symptoms were relieved shortly after eradication therapy in the majority of patients with ulcer disease or erosive lesions. However, significantly smaller portion of the patients with gastroduodenitis became symptom-free. Only about one third of the treated patients remained symptom-free 1 year after the eradication.

Dopaminergic characteristics of isolated parietal cells from rats.

Recently we have identified a dopamine-producing system in the gastric mucosa of rats. All the available morphological data suggest that parietal cells synthesize dopamine. In the present study we investigated the dopaminergic characteristics of isolated parietal cells by different methods. Mixed gastric mucosal cells were isolated and size-fractionated by elutriation. The proportion of neurons, parietal and endocrine cells in the fractions were determined by immunocytochemistry (ICC) using antibodies to neurofilament, proton pump and chromogranin A, respectively. No neurons were found in any of the cell preparations, while 56% parietal cell and 0.0% endocrine cell were achieved in the parietally enriched fraction. By Western blot, a tyrosine hydroxylase (TH, the rate-limiting enzyme of the catecholamine synthesis) immunoreactive protein species was demonstrated in isolated mucosal cells, comigrating with the TH immunoreactivity from PC12 cells. The TH immunoreactivity was colocalized to parietal cells by ICC. Dopamine transporter (DAT), a regulator of extracellular/intracellular dopamine balance in the nervous system, was also demonstrated in parietal cells. A significant amount of dopamine and DOPA were measured by HPLC (13.4 and 9.57 pg/10(6) cell, respectively) in parietally enriched cell fraction. Since this enriched cell fraction was virtually clear of both neurons and endocrine cells, demonstration of TH enzyme, DAT and dopamine in this fraction confirms that the parietal cell population might be a major source of dopamine in the rat stomach, supporting our previous results achieved using whole tissue samples.

Susceptibility of dopamine D5 receptor targeted mice to cysteamine.

BACKGROUND: Recently we demonstrated that gastric mucosa of rats can synthesize, store and release dopamine. Out of five different subtypes, mRNA of D5 (=D1b) dopamine receptor is very abundant in the gastric epithelium. D1 receptor selective dopamine agonists have been shown to protect against experimental gastro-duodenal lesions. AIMS: To test the hypothesis that protective effects of dopamine involve D5 receptors, mucosal lesions were induced in D5 receptor deficient (KO) and wild-type (WT) mice using cysteamine. Morphology and gastric acid secretion of D5 KO mice were also studied. METHODS: Single doses of 600 mg/kg, 300 mg/kg cysteamine or vehicle were administered subcutaneously to fasted animals. After 24 h, number and severity of gastro-duodenal lesions were analyzed. Basal and histamine-induced maximal gastric acid output were measured by a stomach-sac wash-through method. RESULTS: All the KOs in the 600 mg/kg cysteamine group died within 4 h showing symptoms of toxicity while three out of four WTs survived (P<0.05). Mortality after 300 mg/kg cysteamine was significantly higher in KOs versus the WTs: 6/14 versus 2/11, P<0.05. Gastric lesion-index was also significantly higher in KOs (median, middle quartile): four (3-9) versus 0 (0-0), P<0.05. Duodenal lesions did not develop from this single dose of cysteamine in either genotype. Basal and histamine-induced maximal gastric acid output were comparable in the two genotypes. CONCLUSIONS: This study demonstrates that loss of D5 receptor causes mucosal vulnerability and increased toxicity of cysteamine in genetically manipulated mice. Thus, D5 receptor subtype is indeed likely to be involved in protective effects of dopamine in the stomach.

Effects of a novel Hungarian antacid containing Al and Mg (Tisacid) on mucosal prostaglandin generation and oxygen free radicals in normal rats.

A new patented chemical agent (Al-Mg-hydroxy-carbonate; acid-binding capacity greater than 30 mmol/g) was produced by our work-team. After our preliminary pharmacological and some prospective, randomized, multicentre, controlled clinical studies, this antacid was registered (Tisacid tablet and suspension; Alkaloida, Hungary). A cumulative ulcer healing rate of 80-85% was proved by Tisacid monotherapy applied in low doses (from 80 to 160 mmol/day) in patients with duodenal ulcer. The aims of this study were: (i) to determine the role of different antacids on the genesis of mucosal prostaglandins (PGs) (PGE2 and 6-keto-PGF1 alpha) in normal rats; (ii) to evaluate the effects of indomethacin pre-treatment (20 mg/kg b.w.,s.c.) on the Tisacid-induced alterations of gastric mucosal PG-contents; (iii) to analyse the generation of oxygen free radicals and lipid peroxidation in the rat oxyntic mucosa by the application of different doses of Tisacid (activities of CAT, GSH-px and SOD, contents of MDA and red. GSH). It was found that: Tisacid has a potent gastroproprotective effect in gastric mucosa, via (a) an increase in the mucosal levels of PGs, and (b) a scavenging-like effect in normal rat gastric mucosa. It is concluded that the gastroprotective effect of Tisacid appears because of the following: (i) excellent acid-neutralizing capacity; (ii) mucosal generation of PGs (PGE2 and PGI2); (iii) free radical scavenging; (iv) its possible activity as a Ca-antagonist (Mg-containing compound).

[Testing the acid-binding capacity of Tisacid suspension and tablets by intragastric pH determination in patients with hyperacidity]. / Tisacid szuszpenzió és tabletta savkötö képességének vizsgálata intragasztrikus pH-méréssel hiperacid betegekben.

The effect of a Hungarian Al-Mg-containing drug, called Tisacid was studied using of 2.0, 1.0 and 0.5 gram doses. Two administration forms, suspension and tablet were compared. Ten informed, adult hyperacid volunteers were included into the program and three times 5-hour intragastric pH-metry was carried out in a randomized, self-controlled open clinical study (Control, after administration of tablet or suspension). Both forms were found effective in gastric acid reduction at doses of 2.0 and 1.0 g. Tisacid suspension revealed a more pronounced efficacy at the dose of 2.0 g in comparison to the same dose of tablet form. The 0.5 g dose seemed insufficient in reduction of gastric acidity in hyperacid patients applied in tablet or suspension forms. The authors emphasize the role of continuous intragastric pH-metry in clinical practice and investigation of antacids and antisecretory drugs.

[Beneficial effect of the ACE inhibitor captopril in normotensive patients with insulin-dependent diabetes]. / Az ACE-gátló captopril kedvezö hatása normotenziós, inzulindependens cukorbetegekben.

The increase of glomerular filtration can often be observed in patients with insulin dependent diabetes mellitus, even in the early stage of the disease and it does not require the presence of microalbuminuria. This phenomenon can be explained by vasoconstriction occurring in the efferent arterioles. Eighteen normotensive, diabetic patients (aged: 28-42) who developed increased glomerular filtration were recruited in this study. The specific objectives were: 1. to study the beneficial effect of angiotensin converting enzyme inhibitor on the glomerular filtration, 2. to evaluate the effect of this treatment on blood pressure and hemodynamic parameters in normotensive, diabetic subjects. After a placebo period of one week, patients were treated orally a daily dose of 3 x 6.25 mg of captopril for twelve weeks. Glomerular filtration was assessed by the isotopic clearance method and blood pressure recordings were taken every 30 minutes throughout a day using an automatic programmable device. Preload, afterload and linear ejection fraction were estimated by echocardiograph, whereas cardiac index was measured by isotopic first pass technique. At the end of the treatment period a significant decrease of glomerular filtration was observed (from 141.9 +/- 10 ml/min to 98.9 +/- 12 ml/min; p < 0.01. Similarly, the afterload exhibited a significant drop due to drug treatment (45.6 +/- 5.8 x 10(3) dyn/cm2 vs. 55.4 +/- 4.7 x 10(3) dyn/cm2 at the end of the placebo period (p < 0.01). However, preload, linear ejection fraction, and cardiac index did not significantly change during the treatment. According to the results obtained from this study a beneficial effect of captopril on the early development of the glomerular hyperfiltration was demonstrated in normotensive diabetic patients who did not develop microalbuminuria. This issue needs to be investigated further in a large scale clinical trial.

Participation of capsaicin-sensitive afferent nerves in the gastric mucosa of patients with Helicobacter pylori-positive or-negative chronic gastritis.

Capsaicin-sensitive afferent nerves (CSANs) are involved in the protection of gastric mucosa. To clarify the role of CSANs in human Helicobacter pylori-negative or -positive chronic gastritis, after bacterium detection by rapid urease test, (14)C urea breath test, and specific histological staining, the immunodistribution of capsaicin receptor, calcitonin gene-related peptide (CGRP), and substance P (SP) was studied in 21 H. pylori-positive and 30 H. pylori-negative patients with chronic gastritis and 20 patients with functional dyspepsia (as histologically healthy controls). The expression of capsaicin receptor, CGRP, and SP was significantly higher in the mucosa of patients with chronic gastritis than in controls, however, no significant difference was obtained in the immunodistribution in patients with H. pylori-negative versus H. pylori-positive gastritis. In conclusion, CSANs participate in the development of human gastritis, however, their participation does not depend on the presence of Helicobacter pylori as a causative factor.

Gastrointestinal immunology: cell types in the lamina propria--a morphological review.

Defense mechanisms--including immune responses--of the gastrointestinal (GI) system rely on a delicate balance of multidirectional interactions of different components of the GI mucosa. The majority of the cells involved in immune reactions are in the lamina propria (LP) and in the submucosa. Several biologically active substances (enzymes, neurotransmitters, humoral mediators) and their receptors have been reported to be present in LP cells. These cells are in close morphological connections with the surface epithelial cells and with the surrounding vessels and nerve fibers, suggesting a functional association with them. In this paper cell types of the LP will be reviewed from a morphological aspect. In summary, LP cells can be classified as basic structural elements (fibroblasts, fibrocytes, vascular endothelial and smooth muscle cells), blood cells (granulocytes, mast cells, macrophages, T and B lymphocytes, plasma cells), and occasional epithelial and endocrine cells of the surface epithelium. Nerve fibers and terminals, but not neuronal perikarya, can also be seen in the LP. The appearance and the proportion of LP cells strongly depend on the functional activity of the GI system at any given time. Their number and distribution might be significantly altered in certain pathological conditions (infections, inflammations, ulcerations, and other GI disorders). We hope, this review may help clinicians, pathologists, and researchers in the recognition of LP cell types, and in demonstrating their activation, migration and proliferation in different physiological and pathological conditions.

Identification of endogenous peroxidase-containing cells as eosinophils in the gastrointestinal system.

Endogenous peroxidase (EPX) activity in certain cells in the gastrointestinal system interferes with immunohistochemical methods based on the horseradish peroxidase-catalyzed substrate deposition. We studied the distribution and characteristics of these cells. We also report an effective and antigen-preserving EPX blocking method, to make possible the evaluation of immunoperoxidase stainings in cryostat sections. The EPX-containing cells (EPX cells) are present in every part of the gastrointestinal tract, predominantly in the tunica propria. We identified them as eosinophil cells in May-Grünwald-Giemsa stained sections. The complete match was confirmed by different fluorescence techniques. Firstly, the EPX cells were labeled by a red fluorochrome-conjugated substrate of peroxidase enzymes, rhodamine-tyramide, whereas the eosinophil cells were labeled by the green fluorochrome, l-hydroxy-3,6,8-pyrenetrisulfonic acid, which is known to label exclusively eosinophilic granules at pH 10. Secondly, all the EPX cells reacted with a monoclonal antibody against the eosinophil peroxidase enzyme. Finally, a set of commercially available leukocyte markers was used to characterize the EPX cells colabeled by fluorochrome-tyramides. Neither macrophages nor mast cells showed EPX activity. Increased numbers and altered distribution were seen in stressed rats and in ulcerated human stomach.