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Biomedical subjects

J Lechago

Publications and source records attributed to J Lechago.

At least 55 records · Page 3Linked to original sources

Choledochal fungal ball. An unusual cause of biliary obstruction.

A 31-year-old patient with sickle-cell disease who had previous cholecystectomy developed acute onset of jaundice and abdominal pain. An endoscopic retrograde cholangiography demonstrated multiple filling defects within the bile ducts. Microscopic examination of "calculi" removed at surgery revealed that a fungal ball composed of Candida was the cause of biliary obstruction in this case. The patient eventually recovered after removal of the fungal masses and intrabiliary instillation of amphotericin.

Adult↗

Cultured circular smooth muscle from the rabbit colon.

Although cultured vascular smooth muscle cells have been extensively characterized and investigated, there are very few studies of cultured intestinal smooth muscle cells. The aim of this study was to culture colonic smooth muscle (CSM) cells from the rabbit colon. Freshly isolated CSM cells from the circular muscle layer of the distal colon were prepared by collagenase digestion. In primary culture, CSM cells attached to the culture vessels by 48 to 72 h, proliferated by 3 to 7 d, and reached confluency by 14 to 17 d with a "hill-and-valley" pattern. Spontaneous contractions were not observed at any time at 21 degrees or 37 degrees C. Confluent primary cultures were greater than 95% CSM cells, as identified by intensely positive immunofluorescent staining to smooth muscle actin-specific CGA7 and muscle-specific HHF-35 monoclonal antibodies. Transmission electron microscopy of freshly isolated and proliferating CSM cells revealed ultrastructural features consistent with smooth muscle cells. We successfully cultured CSM cells of the rabbit from freshly isolated cells and validated these CSM cells by electron microscopy and immunocytochemical staining. These highly pure primary cultures may be used to investigate numerous aspects of CSM cell metabolism and physiology.

Animals↗

Multiple malignant rectal carcinoid tumors with immunocytochemical demonstration of multiple hormonal substances.

This case report deals with a patient who presented with 17 small carcinoid tumors of the rectum, the largest measuring less than 1 cm in diameter. After a review of the literature on this subject, this case is believed to be highly unusual in several respects: (1) despite their small size, these tumors resulted in multiple metastases to the regional lymph nodes; (2) both primary tumors and metastases were extensively and strongly argentaffin; and (3) most tumors and metastases contained, when investigated by immunocytochemistry, several biologically active substances such as serotonin, somatostatin, and glucagon-like immunoreactivity.

Carcinoid Tumor↗

The endocrine cells of the digestive tract. General concepts and historic perspective.

The chromaffin cells of the digestive mucosa were discovered by Heidenhain in 1870, long before the hormone concept was formulated. Following the discovery of the first hormone, secretin, by Bayliss and Starling in 1902, the study of the cells producing such messengers acquired new impetus. After Masson, in 1914, showed that the digestive chromaffin cells were argentaffin and suggested their endocrine nature, a series of technological advances led to the gradual characterization of diverse cell types, their classification, and the discovery of their function. Histochemistry, including silver impregnations, fluorescence microscopy, and aniline stains, exposed the complexity of the digestive endocrine cells. Electron microscopy provided structural markers for their characterization and the basis for the formulation of the first universally accepted classifications. Light- and electron-microscopic immunocytochemistry contributed to the understanding of the function of many of these cells, thus opening the way for modern classifications. Many unsolved problems still remain. These include the existence of cells without defined function, of chemical messengers without known cell of origin, and the presence of multiple messengers in some cell types. Answers are expected to emerge from further application of immunocytochemistry and from the introduction of modern approaches, such as in-situ hybridization histochemistry.

APUD Cells↗

Immunocytochemical localization of nerve growth factor in mouse kidney.

The presence of NGF in mouse kidney was investigated using immunocytochemical methods. Female and male adult Swiss-Webster mouse kidneys were fixed by perfusion with 4% paraformaldehyde or Zamboni's fixative. The kidneys were frozen, and serial sections were prepared. Rabbit NGF antiserum was used for the primary incubation, and the avidin-biotin complex immunoperoxidase procedure was utilized for immunostaining. NGF immunoreactivity was localized in the apical and perinuclear cytoplasm of cells lining the late distal nephron, in a portion that corresponds to the connecting tubule. This portion of the nephron has been consistently observed in close anatomical relationship with arterioles, which are known to be richly innervated. Some cells of the connecting tubule, corresponding to intercalated cells, lacked NGF immunoreactivity.

Animals↗

Bile duct carcinoma associated with multiple von Meyenburg complexes in the liver.

A 61-year-old white woman was found to have disseminated abdominal carcinoma at autopsy. The primary site proved to be a bile duct carcinoma arising from ductal epithelium of multiple von Meyenburg complexes of the liver. No other neoplastic lesion was found elsewhere after careful, gross, microscopic examination. This case is thought to represent malignant transformation of the hamartomatous growth, previously reported only exceptionally.

Abdomen↗

Effect of acute experimental colitis on rabbit colonic smooth muscle.

The membrane potential and contractile activity of colonic circular smooth muscle from New Zealand White rabbits were studied after the production of acute experimental colitis. Colitis was induced in the distal colon by rectal infusion of formaldehyde solution, followed by an intravenous bolus of soluble immune complexes. Despite active mucosal inflammation, there are only occasional inflammatory cells in the muscularis. Electrophysiological studies on tissue from control rabbits and rabbits with colitis were performed using double sucrose gap and intracellular microelectrode techniques. The resting membrane potential was lower (-44 +/- 3 mV) in muscle from rabbits with colitis compared with control animals (-54 +/- 2 mV) (P less than 0.02). Amplitude of the electrotonic potential after a hyperpolarizing current pulse was decreased (P less than 0.05) and the time constant was shortened (P less than 0.01) in muscle from animals with colitis compared with normal animals. Amplitude (13.1 +/- 2.3 mV) and maximum rate of rise (0.24 +/- 0.06 V/s) of the spike potential, initiated by a depolarizing current pulse, were decreased in muscle from animals with colitis compared with muscle from healthy animals (P less than 0.001). Isometric tension generation after electrical and chemical depolarization of the membrane or bethanechol administration was decreased (P less than 0.001) in muscle from colitic animals. These studies suggest 1) membrane resistance and membrane potential are decreased in muscle strips from animals with colitis; and 2) there is a disturbance in the electrical and mechanical response of these tissues after stimulation.

Action Potentials↗

Immunocytochemical localization of epidermal growth factor in mouse kidney.

Epidermal growth factor (EGF) was originally isolated from mouse submandibular glands (SMG). However, SMG removal failed to lower circulating EGF, and large amounts of EGF have been found in mouse urine. In addition, the presence of pre-pro-EGF mRNA in mouse kidney has recently been reported by others. Kidneys may therefore represent an alternate source of EGF. In the present study, we investigated the immunocytochemical localization of EGF in mouse kidney. Male and female adult Swiss Webster mice were fixed by perfusion with 4% paraformaldehyde or Zamboni's fixative, the kidneys were frozen, and serial sections were obtained. Rabbit EGF antiserum was used for the primary incubation and the avidin-biotin complex immunoperoxidase procedure was utilized for immunostaining. EGF was immunolocalized in the apical portion of the cells lining the thick ascending limb of Henle (TALH) and the distal convoluted tubule (DCT). The macula densa, in contrast, lacked EGF immunoreactivity. No sex differences were observed in the distribution pattern or intensity of immunostaining. Infusion of EGF into sheep renal artery has been reported to induce changes in urine flow and ionic composition. Immunolocalization of EGF in the TALH and DCT documented here supports a regulatory role for EGF in the function of the mouse distal nephron.

Animals↗

Two biochemically distinct populations of histaminocytes separated by isokinetic sedimentation of dispersed rat gastric cells.

Two populations of histaminocytes, with different sedimentation rates (SR), were separated by a computer developed isokinetic gradient using dispersed rat gastric mucosal cells. Histamine content, histidine decarboxylase (HDC) activity and incorporation of radiolabelled histidine metabolites were used to assess the migration of specific cells throughout the gradients. One histaminocyte population, with cells of lower SR, contained high HDC activity and undetectable levels of histamine, whereas the other population, with cells of higher SR, contained lower HDC activity and high concentration of histamine. Both types of histaminocytes incorporated 3H-histidine metabolites. Electron microscopy showed that the fractions containing histaminocytes with lower SR had 3.5 times more endocrine ECL cells than the original population of dispersed fundic cells and lacked A and D cells, whereas the fractions with histaminocytes of higher SR were associated with a 2.7 times higher concentration of A and D cells and with a 7.7 times higher ratio of a variety of partial cells with a distinct mitochondrial morphology. These results are consistent with prior novel information regarding the separation of two populations of rat histaminocytes using different sedimentation techniques.

Animals↗

Immunocytochemical localization of a granuliberin-like peptide in Rana pipiens brain.

The histamine-releasing peptide "granuliberin", originally isolated from the skin of Rana rugosa, was localized by immunofluorescence within nerve cell bodies and fibers in the brain of Rana pipiens. The granuliberin-positive neurons were characterized ultrastructurally by electron microscopic observation of ultrathin sections stained either with immunoperoxidase or with conventional stains. Granuliberin-positive nerve cell bodies were seen throughout the hypothalamus, from the suprachiasmatic area rostrally to the full length of the periinfundibular grey matter caudally. Similarly positive nerve fibers were localized in the hypothalamus radiating upwards to the optic vesicles in the midbrain, extending through the preoptic area into the subpallium in the forebrain, and throughout the white matter surrounding the floor and lateral walls of the fourth ventricle in the brainstem. The granuliberin-positive nerve cells showed the presence of variable numbers of small cytoplasmic neurosecretory granules, possessing an electron dense elongated core, and measuring 250-350 nm in their largest diameter. The functional significance of a granuliberin-like peptide in neurons of the frog brain is not known.

Animals↗

Congenital pernicious anemia: report of seven patients, with studies of the extended family.

Seven children ages 1 1/2 to 12 years with congenital pernicious anemia were detected in an extended Mexican family. All affected children had megaloblastic anemia accompanied by low serum B12 and normal serum folate levels. Gastric fluid analysis in six patients revealed normal gastric acidity and absent intrinsic factor. Serum antibodies to intrinsic factor or parietal cells were also absent. Schilling tests performed in six of the seven patients yielded abnormal results. Of the three patients in whom gastric biopsy was done, two had normal histologic findings (including examination by electron microscopy) and one had mild atrophy. All patients responded rapidly to parenterally administered vitamin B12 therapy. In addition, 170 family members were screened for the defect with complete blood counts and serum B12 levels. Such screening detected pernicious anemia in two of the children, but no other abnormalities that could be attributed to pernicious anemia were found in other family members. Based on the family pedigree, autosomal recessive inheritance is likely. The variability of age of presentation in this family is noteworthy and suggests that expression may be modified by still undefined factors.

Anemia, Pernicious↗

Immunohistochemical studies in endocrine carcinoma of the skin.

Twenty-two endocrine carcinomas of the skin were examined immunohistochemically for the presence of several polypeptides and serotonin. Eight tumors contained one or more polypeptides. Occasional cells containing calcitonin were found in several tumors. Rare cells with substance P, somatostatin, and ACTH were present in only one tumor each. None of the patients presented a clinical syndrome associated with any of these polypeptides. Met-enkephalin and serotonin were negative in all tumors. The lack of demonstration of met-enkephalin in these tumors makes their relationship to the Merkel cell uncertain.

Adenocarcinoma↗

Transient hypergastrinemia of 2 years' duration in a young pediatric patient.

A 24-month-old female child experienced watery diarrhea, growth failure, and abdominal pain from age 3 months. Hypergastrinemia, hypochlorhydria, and fundic gastritis were documented. A secretin stimulation test was normal but protein meal stimulation revealed an abnormal serum gastrin response. Antral biopsies revealed G cell hyperplasia. Chronic treatment with antacids and an anticholinergic agent was unsuccessful. Spontaneous recovery occurred at age 29 months. Gastrin stimulation tests, gastric acid secretory tests, antral mucosal biopsies, and multiple basal serum gastrin levels were repeated. All were normal. Follow-up of greater than 3 years has documented a completely normal clinical and laboratory course.

Achlorhydria↗

Human colonic substance P-producing cells are a separate population from the serotonin-producing enterochromaffin cells.

Human colonic mucosa was immunostained with antibodies against substance P to identify the endocrine cells containing this peptide in the mucosal glands. Dual immunohistochemical and histochemical studies were also carried out to determine whether these cells are enterochromaffin cells and contain serotonin as claimed in the literature. The results obtained indicate that the normal human colonic substance P-producing cells are not argentaffin cells, nor do they contain serotonin. In addition, they are also negative both for several silver and for other techniques commonly used to identify digestive endocrine cells. They are positive, however, with the argyrophilic technique of Churukian-Schenk. It is concluded, therefore, that the substance P-producing cells of the human colonic mucosa are not a subpopulation of the enterochromaffin cells, but constitute a distinct and independent cell type.

Colon↗

Vasoactive intestinal polypeptide-immunoreactive nerves in the kidney.

Nerve fibers immunoreactive for vasoactive intestinal polypeptide (VIP) were demonstrated by immunocytochemistry in the dog and rat kidney. They were seen in association with the renal artery and its branches. In the dog, VIP-immunoreactive fibers were rarely seen close to small blood vessels suggestive of arterioles. The possible existence of neuroeffector junctions between VIP-positive fibers and renin-secreting juxtaglomerular cells requires further investigation. VIP-positive renal nerves, however, might have a vasodilatatory role.

Animals↗

Presence of ranatensin-like and bombesin-like peptides in amphibian brains.

Antibodies specific for the carboxyl-terminal regions of bombesin and of ranatensin were used to study the tissue distribution of substances immunochemically similar to these two peptides in three amphibian species. Brain extracts of Rana catesbeiana, Rana pipiens, and Xenopus laevis all contained considerable quantities of both types of immunoreactivity, with measured concentrations as high as several hundred pmol per g tissue. The two antibodies used in this study had very low crossreactivity (less than 1% by RIA) with the other peptide. In addition, gel filtration revealed different elution profiles for the two immunoreactive substances extracted from amphibian brains. Immunocytochemistry revealed differences in localization within nerve fibers and cell bodies and specific absorption by the appropriate peptide. Ranatensin-like peptides were also present in high concentrations in skin of the two Rana species but not in that of Xenopus. Bombesin-like peptides were more abundant in the stomach of all three species. Significant amounts of substance P/phylasaemin-like immunoreactivity also were detected in the brains of all three species. It is concluded that ranatensin-like peptides are not confined to the skin and can be included as central nervous system neuropeptides in amphibians. These two groups of peptides are not species-specific since both are found in brain and stomach of amphibians whose skin contains only ranatensin-like peptide or neither.

Animals↗

Innervation of rat antral gastrin-producing cells.

The innervation of rat antral gastrin-producing cells (G-cells) was studied by light and electron microscopy. Combination of histochemistry for acetylcholinesterase and immunofluorescence for gastrin in the same tissue section showed apparent contact between some of the G-cells and acetylcholinesterase-positive nerves. Electron microscopic observation, however, revealed gaps of 200-500 nm or more between the G-cells and the closest nerve axons which often contained large dense-cored vesicles. The latter may represent the storage sites for neuropeptides previously localized by immunohistochemistry in gastric nerves.

Acetylcholinesterase↗