Search PubMed⌕ Search

Biomedical subjects

A Gerolami

Publications and source records attributed to A Gerolami.

At least 19 recordsLinked to original sources
PubMed1997

NHE-3 isoform of the Na+/H+ exchanger in human gallbladder. Localization of specific mRNA by in situ hybridization.

BACKGROUND/AIMS: Electroneutral absorption of NaCl by the gallbladder mucosa is likely to depend at least in part on a Na+/H+ exchanger. In intestine and colon, absorption due to Na+/H+ exchanger is explained by the presence of specific isoforms of the exchanger, the NHE-3 isoform and possibly the NHE-2 isoform. The aim of the present work was to determine whether the mRNAs coding for NHE-2 and NHE-3 are expressed in epithelial cells of human gallbladder. METHODS: Total RNAs from human gallbladder were subjected to reverse transcription-polymerase chain reaction using specific primers. No message was observed with NHE-2 specific primers, showing that NHE-2 isoform plays no role in gallbladder absorption. With NHE-3 specific primers, a 239 bp cDNA fragment was obtained and showed a high homology with the NHE-3 isoform, confirming the presence of NHE-3 in the gallbladder wall. This fragment was cloned in a pLitmus vector in order to produce cRNA probes by in vitro transcription. Cellular localization of the NHE-3 mRNA was studied on cryostat sections using the cRNA probes labeled with Digoxigenin-11-UTP, controls included assays with sense probe, antibodies without probe and RNaseA treated tissue. A specific staining of the NHE-3 mRNAs was found to be strictly localized to the gallbladder epithelial cells. RESULTS/CONCLUSIONS: Expression of NHE-3 in the gallbladder was found only in the absorptive epithelial cells. The NHE-3 isoform of the Na+/H+ exchanger is likely to be involved in water and electrolyte absorption from bile.

Base Sequence↗
PubMed1996

Role of the NHE3 isoform of the Na+/H+ exchanger in sodium absorption by the rabbit gallbladder.

The absorption of water and electrolytes by the gallbladder seems to be largely dependent upon a Na+/H+ exchange at the apical membrane of the gallbladder epithelium. To find out if the exchanger involved is the NHE3 isoform, as in other absorbing epithelia, two studies were performed using the rabbit gallbladder. First, we studied 22Na absorption in Ussing chambers with Krebs buffer as a control solution, and in the presence of amiloride (100, 200 or 1000 microM), ethyl-isopropyl-amiloride (EIPA, 1 or 5 microM), or the phorbol ester, phorbol 12-myristate 13-acetate (PMA, 1 microM). A net mucosal-to-serosal Na+ flux was observed with control buffer. No inhibition of this net flux was observed with 5 microM EIPA, and the IC50 for amiloride was found to be 200 microM. PMA induced a reduction of absorption by 30% that was prevented by incubation with calphostin C. Resistance to amiloride and EIPA, and inhibition by PMA are consistent with the involvement of the NHE3 isoform. The second study involved reverse-transcriptase polymerase chain reaction (RT-PCR) of total gallbladder RNA, with two primers designed to amplify a 645-base-pair fragment from NHE3 mRNA. A cDNA fragment of the expected size was actually obtained from gallbladder RNA, while RT-PCR of RNA from the liver, which does not contain NHE3, gave negative results. A sequence of 492 nucleotides of the amplified product was determined, which was almost superimposable onto the known sequence of the corresponding fragment of rabbit NHE3. It is concluded that, in rabbit gallbladder, neutral NaCl absorption is, at least in part, dependent on the NHE3 isoform of the Na+/H+ exchanger.

Absorption↗
PubMed1996

Inhibition of gap junction intercellular communications of cultured rat hepatocytes by ethanol: role of ethanol metabolism.

BACKGROUND/AIMS: In a previous study, we reported that in cultured rat hepatocytes, ethanol inhibits intercellular communication which is known to play a central role in the regulation of cell growth and differentiation. This work was designed to find out if ethanol exerts a direct action on cell membranes, comparable to other long-chain (C6-C9) alcohols, or an indirect action. METHODS: Intercellular communication was measured on short-term cultured rat hepatocytes by the fluorescent Lucifer-Yellow CH transfer method. Intracellular pH was measured by spectrofluorimetry and membrane expression of connexin 32 by indirect immunofluorescence. RESULTS: Under our conditions, ethanol (20 mM) inhibited intercellular communication of hepatocytes to the same extent as did octanol and 1 mM. Immunofluorescence semi-quantitative studies of connexin 32 suggested that the observed inhibition was not related to a decrease in the number of gap junction plaques. In contrast with those of octanol, the inhibitory effects of ethanol appeared to be indirect because the inhibition of ethanol metabolism by 4-methyl pyrazole abolished its effects on intercellular communication, while 4-methyl pyrazole did not influence the effects of octanol. Acetaldehyde, the main metabolite of ethanol was without effect on gap junctions. CONCLUSIONS: This suggests that the inhibition of intercellular communication induced by ethanol may be included among the consequences of intermediary cell metabolism disturbances indirectly due to ethanol oxidation. This may be one of the mechanisms by which ethanol metabolism exerts a hepatotoxic possibly carcinogenic action.

1-Octanol↗
PubMed1996

Lack of interaction between hepatitis C virus and alcohol in the pathogenesis of cirrhosis. A statistical study.

BACKGROUND/AIMS: In several studies markers of hepatitis C virus infection have been shown to be present in alcoholic patients with cirrhosis. Our work was designed to test the likely hypothesis that this association is due to an interaction between hepatitis C virus and alcohol in the pathogenesis of cirrhosis. METHODS: We compared alcohol consumption and repartition of anti-HCV antibodies detected by an immunoblot recombinant assay in 101 male patients with cirrhosis and in 120 male controls. Interactions between anti-hepatitis C virus, alcohol and cirrhosis were calculated using log linear hierarchical models for frequency data. The basis of the method is that an interaction between hepatitis C virus and alcohol implies that a model built on the hypothesis of a role of hepatitis C virus and alcohol in the disease should be improved by a coefficient associated with multiplicative effects of hepatitis C virus and alcohol. RESULTS: In patients with cirrhosis the mean alcohol consumption (148 +/- 100 g per day) and the incidence of positivity for anti-HCV antibodies (45%) were significantly higher than in controls. The results were consistent with a theoretical model built with the hypothesis of an independent role of both alcohol and hepatitis C virus. The goodness of fit between this model and the actual distribution of alcohol consumption and hepatitis C virus markers was not improved by introduction of an interaction between hepatitis C virus and alcohol. CONCLUSIONS: In alcoholic subjects with hepatitis C virus infection, the probability to have cirrhosis seemed to be explained by additive effects of alcohol and hepatitis C virus. From a purely statistical point of view, no interaction between hepatitis C virus and alcohol consumption on a multiplicative scale could be demonstrated.

Aged↗
PubMed1996

Presence of the NHE3 isoform of the Na+/H+ exchanger in human gallbladder.

1. In man and in various animal species, absorption of NaCl from bile by the gallbladder mucosa is associated with luminal proton secretion. A similar absorption of NaCl in small intestine, colon and renal tubule is related, at least in part, to the presence of the NHE3 isoform of the Na+/H+ exchanger. This work was designed to find out whether NHE3 is also present in human gallbladder. 2. At surgery, 100-200 mg of the gallbladder wall was obtained from patients treated by cholecystectomy for gallstones. After isolation of the mucosa, total RNA was extracted and submitted to reverse transcription-polymerase chain reaction with two primers: 5'-AAGCCICTGGTGCAGTGGCTGAAGG-3' and 5'-GGAGTCCTTIAAGTCGGCIAAGCTGGGC-3', designed to amplify a sequence of 645 bp of rabbit NHE3 mRNA (642 bp in man). RNA from human liver and from rabbit heart, neither of which contain NHE3, and human ileal RNA, which does contain NHE3, were used as controls. 3. RNA extracted from the mucosal moiety of the gallbladder wall gave an amplification product of about 645 nucleotides. Controls gave the expected negative or positive results. Sequencing of the amplified RNA showed it was almost identical to previously determined sequences of NHE3 in other human tissues. 4. It is concluded that the mucosa of human gallbladder contains the mRNA of NHE3 isoform. This isoform could therefore play a role in sodium absorption from bile.

Animals↗
PubMed1995

Evidence for apical Na+/H+ exchanger in bovine main pancreatic duct.

The finding of a high PCO2 in basally secreted pancreatic juice of man and dog raises the hypothesis of proton secretion from ductal epithelial cells presumably through a Na+/H+ exchanger. To test this possibility, H+ luminal secretion and Na+ movements were measured in vitro on samples of bovine pancreatic ducts mounted in Ussing-type chambers. The rate of luminal acidification measured by the pH stat method, using bicarbonate-free media gassed with 100% O2, reached 2.75 muEq/cm2/hr. Proton secretion was blocked in the presence of 1 nM amiloride or in the absence of Na+ (replaced by choline) in the mucosal solution. Study of transepithelial 22Na fluxes in short-circuited tissue, bathed on both sides by control Ringer solution, gassed by 95% O2-5% CO2 demonstrated a net sodium transport from the mucosal to the interstitial side of the duct (net 22Na flux = 3.23 +/- 0.8 muEq/cm2/hr). This net sodium transport was electroneutral and blocked by mucosal amiloride (0.5-1 mM/liter) or by interstitial ouabain (1 mM/liter). These results are consistent with the existence of a Na+/H+ exchanger on the luminal side of the bovine main pancreatic duct.

Amiloride↗
PubMed1994

[Hepatic granulomatosis associated with mononucleosis syndrome secondary to cytomegalovirus infection: apropos of 2 cases in healthy adults].

In this study, the authors report two observations of granulomatous hepatitis. The secondary appearance of a mononucleosis syndrome, three weeks after the onset of fever, in healthy adults, evoked the diagnosis of a cytomegalovirus infection. The authors insist on the histologic and virologic differences of the CMV infection between the healthy adults and the immunodepressed patients. They also note the difficulties of the diagnosis of the CMV infection in healthy adults.

Adult↗
PubMed1994

Influence of hypercalcemia on ionized calcium concentration in pancreatic juice of the dog.

Calcium concentration of pancreatic juice depends on secretion of calcium bound to enzymatic proteins or calcium diffusion from interstitial fluids. To evaluate the relative magnitude of these pathways, we studied the influence of hypercalcemia on ionized calcium (Ca++) in dog pancreatic juice. Pancreatic juice was collected during basal secretion and during stimulation by secretin or secretin plus caerulein in control conditions and under CaCl2 infusion. [Ca++] was measured by selective electrodes. Saturation of juice in CaCO3 was calculated. In stimulated juice, total calcium concentration ([CaT]) and [Ca++] were unchanged by hypercalcemia. In basal juice, composition was profoundly modified by hypercalcemia because [CaT] (3.31 +/- 0.89 mmol/L vs 1.80 +/- 0.44 for controls), [Ca++] (1.44 +/- 0.37 mmol/L vs 0.84 +/- 0.24 mmol/L for controls), and the index of saturation in CaCO3 (5.2 +/- 2.4 vs 2.9 +/- 1.8 for controls) increased significantly. Protein concentration was unchanged. This suggests that in basal conditions, the relationship between plasma and juice calcium levels is due to passive interstitial Ca++ diffusion through the pancreatic ducts. In accordance with the hypothesis of a restricted calcium diffusion, the effects of hypercalcemia were flow rate dependent, being less pronounced when basal flow rate increased. It is concluded that, in the dog, the calcium species found in stimulated juice result from a redistribution of calcium secreted along with proteins, whereas at low secretion rate, juice calcium level depends mainly on interstitial Ca++ diffusion into the main pancreatic ducts.

Analysis of Variance↗
PubMed1993

Influence of pancreatic ducts on saturation of juice with calcium carbonate in dogs.

In several species, bicarbonate and calcium concentrations of pancreatic juice are known to vary during the different phases of pancreatic secretion. The effects of these variations on the saturation of juice with calcium carbonate, a critical factor for the formation of pancreatic stones, are not known. In this work, we studied the saturation degree of pancreatic juice with calcium carbonate in six unanesthetized beagle dogs equipped with Thomas cannulae during basal secretion and after bolus injections of cerulein (30 ng/kg) or secretin (0.25 units/kg). In the different samples of pure pancreatic juice, pH, PCO2, bicarbonate, and proteins were measured by standard methods. Total calcium (CaT) and ionized calcium (Ca2+) were determined using calcium-specific electrodes. Saturation with calcium carbonate was calculated by reference to the solubility product of calcite at 37 degrees C. Almost all the samples were found to be supersaturated with calcium carbonate but large variations of the saturation index were observed. In basal samples, obtained during periods of low secretion rate, the mean saturation index (3.35 +/- 3.01) was significantly lower than under secretion (12.10 +/- 5.14) or cerulein (18.01 +/- 8.42). This low basal saturation index, in spite of a high Ca2+ content, was explained by a low bicarbonate concentration (37.6 +/- 18.9 mmol/liter) and a high PCO2 (13.4 +/- 7.5 kPa). In contrast, in juice obtained after hormonal stimulation, PCO2 (4.8 +/- 1.6 kPa) was similar to plasma PCO2 (5.5 +/- 1.2 kPa).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

[Effect of a low-fat diet on the fasting volume and postprandial emptying of the gallbladder].

Little is known of the effects of meal composition on gallbladder emptying and cholecystokinin (CCK) release in man. Gallbladder volumes (measured by means of real time ultrasonography) and plasma CCK levels (determined by radioimmunoassay) were studied in 5 normal subjects, before and after a normal solid-liquid meal, and before and after a low-fat, low-protein, solid-liquid meal after 3 days regimen with low-fat, low-protein meal. Low-fat, low-protein regimen significantly increased gallbladder fasting volume and significantly decreased fasting plasma CCK levels. This suggests that CCK secretion regulates fasting gallbladder volume and that basal CCK secretion depends on diet composition. After a normal meal, gallbladder emptying was biphasic with a 44% volume decrease within the first 15 min followed by slower emptying during the next 60 min with a final volume reaching less than 15% of the fasting volume. After a hypolipidic, hypoproteic meal, the initial 15 min emptying (42%) was the same as after control meals but no further decrease of gallbladder volume was observed. This study shows that the initial phase of post prandial gallbladder contraction is not dependent on meal composition which affects late gallbladder emptying only. We conclude that a low-fat, low-protein diet, increasing gallbladder fasting volume and decreasing gallbladder emptying, may favor gallbladder stasis and therefore increase the risk of gallstone formation.

Cholecystokinin↗
PubMed1992

Effects of ethanol on intercellular communications and polarization of hepatocytes in short-term culture.

The formation of intracellular lumina with apical differentiation is observed in several cancerous epithelial cell lines including human hepatocarcinoma. This disorder of cell polarization can be induced by the inhibition of cell-cell communication, a known factor of carcinogenesis. This work was designed to study the effects of ethanol on the differentiation of hepatocytes in short-term culture. Isolated hepatocytes were plated on plastic culture dishes that were 35 mm in diameter (10(6) cells/dish). Three hours after plating, the hepatocytes were incubated in the presence of 20 mmol/L ethanol for 1 hr. Treated cells were compared with controls using morphometric methods after conventional treatment for ultramicroscopy and by measuring cellular dye coupling by the fluorescent Lucifer Yellow CH transfer method. Bile canaliculi formation decreased in alcohol-treated cells (6.5% vs. 9.9%, 2p less than 0.05), whereas intracellular lumina incidence increased (3.1% vs. 0.5%, 2p less than 0.01). In parallel, the dye-coupling capacity decreased significantly when hepatocytes were treated with alcohol (2p less than 0.01). This work shows that short-term ethanol treatment induces significant disturbances of cell polarization and inhibits the reestablishment of cell-cell communication in cultured hepatocytes. These disorders could, at least in part, explain the carcinogenic effects of ethanol.

Animals↗
PubMed1989

Intracellular lumina and bile canaliculi in rat hepatocytes in vitro--a cytochemical study.

Within a few hours after being placed in culture, isolated rat hepatocytes reassociate into clusters and differentiate intercellular cavities bordered by junctional complexes. These structures bear a strong resemblance to the bile canaliculi found in the liver. If the plating is impaired by permanent agitation, intracellular lumina surrounded by microvilli appear in the cytoplasm of some isolated hepatocytes. The formation of these structures which mostly contain an osmiophilic substance is enhanced by cAMP and they probably result from modifications in the functioning of the Golgi complexes as suggested by thiamine pyrophosphatase cytochemistry. It may be speculated that these intracellular lumina may be functionally related to new bile canaliculi differentiated between reassociated cells.

Animals↗
PubMed1989

Calcium carbonate saturation in human pancreatic juice: possible role of ductal H+ secretion.

Saturation with calcium carbonate was measured in human pancreatic juice anaerobically collected for diagnostic purposes in 15 patients who were ultimately found not to have pancreatic disease. Bicarbonate, PCO2, proteins, and total and ionized calcium were measured in samples collected every minute during a 20-min period after intravenous administration of secretin (1 U/kg) and, 10 min later, caerulein (75 ng/kg). All samples were supersaturated with calcium carbonate. The first sample (washing-out sample) contained the highest concentrations of ionized calcium (Ca2+) and proteins but was least supersaturated with calcium carbonate. Washing-out samples also differed significantly from samples under caerulein by having a lower pH (7.52 +/- 0.30) and higher PCO2 (103.1 +/- 32.20 mmHg) versus 8.22 +/- 0.15 and 38.4 +/- 4.5 mmHg, respectively. Values of PCO2 increased and bicarbonate concentration decreased linearly with protein concentration in washing-out samples but not under hormonal stimulation. This suggests that a concentration of pancreatic juice occurs in pancreatic ducts in association with luminal H+ secretion and CO2 formation, which results from bicarbonate neutralization. During stimulation, ionized calcium concentration increased with protein concentration without any change of PCO2, so that supersaturation was more pronounced under caerulein than under secretin stimulation. Disturbances of the ductal concentration of pancreatic juice during interdigestive periods could be important for pancreatic stone formation in humans.

Bicarbonates↗
PubMed1988

Influence of lipid-rich diets on intestinal cholesterol uptake.

The purpose of this work was to determine whether or not cholesterol transport by intestinal brush border is influenced by fat-enriched diets and therefore plays a role in the regulation of cholesterol absorption. This study was carried out on rats divided into three groups according to diet: (1) control with a diet containing 1.2% cholesterol (diet T), (2) diet T plus 28% saturated fat (lard) and (3) diet T plus 28% polyunsaturated fat (corn oil). Uptake of cholesterol and oleic acid from defined mixed micellar solutions was studied on two experimental models: everted sacs and brush border vesicles of the intestinal membrane. In vivo cholesterol absorption was measured by the dual isotope plasma ratio method of Zilversmit. Fat-enriched diets decreased both in vivo cholesterol absorption and in vitro cholesterol uptake without any specific effect of unsaturated fats. This suggests that the mechanisms involved in the transport of cholesterol across the brush border membrane may be rate-limiting for cholesterol absorption. Oleate and butyrate uptakes, in contrast, were unaffected by the fat content of the diet.

Animals↗
PubMed1988

[Intracellular lumen in cultured hepatocytes of rats. Study of an original cytoplasmic infrastructure].

A few hours after plating, isolated rat hepatocytes reassociate into clusters and differentiate intercellular cavities bordered by junctional complexes. These structures show a great resemblance to bile canaliculi seen in vivo. Intracellular lumens surrounded by microvilli are observed in the cytoplasm of some cultured hepatocytes. The formation of these structures, which contain an osmiophilic substance, probably results from modifications in the functioning of the secretory apparatus. It can be speculated that these intracellular lumens may contribute to the formation of new canaliculi differentiated between reassociated cells.

Animals↗