Growth factor receptors gene expression and Akt phosphorylation in benign human thyroid nodules are unaffected by chronic thyrotropin suppression.

Levothyroxine (L-T4)-based suppression of thyrotropin (TSH) secretion is widely used to prevent the growth of benign thyroid nodules, although the effectiveness of this approach has been demonstrated only in a subset of patients. In this study, we analyzed the in vivo effects of L-T4-mediated TSH suppression on elements of insulin/IGF-1-dependent growth-regulating pathways in tissues from patients with benign thyroid nodules. Nodular and non-nodular tissue specimens were collected from 63 patients undergoing thyroidectomy. 32 had received preoperative TSH suppressive therapy with TSH levels consistently below 0.5 mU/l (L-T4 group). TSH suppression had not been used in the other 31, and their TSH levels were normal (0.8-4 mU/l (control group). Quantitative RT-PCR was used to measure mRNA levels for TSH receptor, IGF1, IGF-1 receptor, insulin receptor, insulin receptor substrate 1 in nodular and non-nodular tissues from the 2 groups. Akt and phosphorylated Akt protein levels were detected by Western blot. Mean levels of mRNA for all genes tested were similar in the 2 groups, in both nodular and non-nodular tissues. The 2 groups were also similar in terms of phosphorylated Akt protein levels (measured by densitometric scan in 10 randomly selected nodules from each group). This is the first demonstration based on the study of human thyroid tissues that TSH suppression does not affect the expression of components of the insulin/IGF-1-dependent signaling pathways regulating thyrocyte growth. This may explain the lack of effectiveness of TSH-suppressive therapy in a substantial percentage of benign thyroid nodules.

Embryonic chick intestine in organ culture. A unique system for the study of the intestinal calcium absorptive mechanism.

Duodena from 20-day-old chick embryos can be maintained in large scale organ culture on specially designed stainless-steel grids in contact with serum-free medium for 48 h with excellent preservation of mucosal structure at both the light and electron microscope levels. Although mitotic rate was subnormal, several other factors attest to the essential viability of the cultured intestine: L-leucine incorporation into protein, as well as the synthesis of a specific vitamin D(3)-induced calcium-binding protein (CaBP), increased over a 48-h culture period, and the electropotential gradient across the intestine was maintained throughout the culture period as was a concentration gradient for calcium. The tissue responded to vitamin D(3) in the medium by synthesizing the calcium-binding protein within 6 h and by exhibiting enhanced (45)Ca uptake within 12-24 h. Concentrations of vitamin D(3), or its 25-hydroxylated derivative, higher than necessary for CaBP induction, also increased the activity of alkaline phosphatase. The 1,25-dihydroxylated derivative of vitamin D(3), at a level extremely potent in CaBP induction, did not stimulate alkaline phosphatase. Mucosal to serosal transport of (45)Ca could also be measured in everted duodenal sacs, subsequent to culture under similar conditions, and was also increased by vitamin D(3) in the medium. Other embryonic organs, esophagus, stomach, liver, pancreas, lung, skin, and muscle, did not produce CaBP in response to vitamin D(3) in the culture medium. However, CaBP-synthesizing capacity was present in the entire intestinal tract, exclusive of the rectum. (59)Fe and (32)P uptake by cultured duodenum were also stimulated by vitamin D(3). The system has proven quite useful in the study of the vitamin D-mediated calcium absorptive mechanism but should be applicable to the study of the absorption of other nutrients, drugs, hormones, etc., as well as other studies of intestinal function.

Embryonic chick intestine in organ culture: response to vitamin D 3 and its metabolites.

Embryonic chick intestine maintained in organ culture responded to vitamin D(3) and its metabolites 25-hydroxycholecalciferol and 1,25-dihydroxycholecalciferol by synthesis of calcium-binding protein and enchanced calcium-45 uptake. The dihydroxy metabolite was by far the most potent inducer of the protein and also acted more rapidly than vitamin D(3) to stimulate isotope uptake. Despite its lower potency, vitamin D(3) itself was effective.

Vitamin D3: induction of calcium-binding protein in embryonic chick intestine in vitro.

Induction of the synthesis of calcium-binding protein in chick embryonic intestine maintained in vitro was accomplished by simply adding vitamin D(3) to the culture medium. Accompanying the induction of this protein, there was enhanced radiocalcium uptake by the intestine. These observations represent the first demonstration of an in vitro physiological effect of vitamin D(3) on the calcium absorptive mechanism of the intestine.

Role of vitamin D glucosiduronate in calcium homeostasis.

Evidence has been presented suggesting the presence of vitamin D(3) 3beta-glucosiduronate and 1,25-dihydroxyvitamin D(3) glucosiduronate in rat bile. To evaluate the role of vitamin D glucosiduronates in calcium and phosphorus homeostasis, we synthesized vitamin D(3) 3beta-glucosiduronate and tested its biological activity in calcium- and vitamin D-deficient rats. After the intravenous administration of vitamin D(3) 3beta-glucosiduronate to rats maintained on a low calcium diet, there was an increase in duodenal calcium transport and an increase in serum calcium. Vitamin D(3) 3beta-glucosiduronate, however, was less active than equimolar amounts of vitamin D(3). At doses of less than 0.65-1 nmol per rat, the conjugate exhibited no activity. When vitamin D(3) 3beta-glucosiduronate was administered to vitamin D-deficient rats, 25-hydroxyvitamin D was detected in the serum; the increase in serum 25-hydroxyvitamin D levels was less than that observed after the administration of an equimolar amount of vitamin D(3). Vitamin D(3) 3beta-glucosiduronate showed no detectable activity in the induction of calcium binding protein in chick embryonic duodena, a system in which no endogenous steroid beta-glucuronidase activity is detectable. These data demonstrate that vitamin D(3) 3beta-glucosiduronate is biologically active in vivo and that the observed activity is due to hydrolysis of the conjugate to vitamin D(3). As vitamin D(3) 3beta-glucosiduronate is excreted in the bile of rats, it is possible that this conjugate is reutilized in vivo after hydrolysis to free vitamin D(3). These results suggest the existence of a mechanism for reutilization of the biliary products of vitamin D(3).

[On the job training at construction sites of the hospital corporation of Careggi (Florence)]. / Tutoraggio formativo in itinere nei cantieri edili dell'Azienda Ospedaliera di Careggi (Firenze).

The "Careggi Safety" project has like objective to guarantee the safety execution of Careggi Hospital (Florence) restoration and development works, thanks to an in itinere training program into building sites and through tutor figure using. The project aim is to overcomes traditional indoor training limits, not effective in complex and dynamics reality like as building sites, constrained by contracts deadline and high labour turnover (subcontracts) inside carry out process. Solutions chose are: (a) a training projected in itinere, following site works evolution and safety and coordination plan, and through a constant agreement between customer and operative enterprises; (b) a building site's tutor, standing beside workers during realization phases, contributing to form on respective safety carry out job. In to "Careggi Safety" project training has been chose as preliminary and obligatory condition for labours admission and control into the building site.

Binding proteins from animals with possible transport function.

Several proteins from various animal tissues with possible transport function have been briefly described, with emphasis given to a vitamin D-induced calcium-binding protein (CaBP) implicated in calcium translocation across epithelial membranes. The latter protein was shown to be present in the small intestine, colon, kidney, and the uterus (shell gland) of the chicken. CaBP was also found in the small intestine of the rat, dog, bovine, and monkey. This protein has been isolated in high purity from chick intestinal mucosa and some of its properties determined. Its molecular weight is about 28,000, its formation constant, about 2.6 x 10(5) M(-1), and its binding capacity, 1 calcium atom per protein molecule. Correlative studies have shown that CaBP concentration in intestinal mucosa varies with the calcium absorptive capacity of the gut, thereby suggesting that CaBP is intimately involved in the process of calcium absorption. CaBP has been localized in the brush border region of the intestinal absorptive cell and within goblet cells. Among other proteins mentioned were the intrinsic factor required for vitamin B(12) absorption and the protein(s) associated with iron translocation.

Positive cotranscriptional regulation of intestinal calbindin-D28K gene expression by 1,25-dihydroxyvitamin D3 and glucocorticoids.

The steroid hormone 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] induces expression of the gene encoding calbindin-D28K, a protein involved in intestinal Ca2+ transport. Glucocorticoids stimulate intestinal development and function, and presumed interaction with 1,25-(OH)2D3 has been intensively studied. Most studies involved administration of high doses of glucocorticoids in vivo, which inhibits intestinal Ca2+ transport by an unknown mechanism. However, it is now known from studies of the duodenal organ culture model that low concentrations of glucocorticoids enhance 1,25-(OH)2D3-dependent calbindin-D28K biosynthesis and Ca2+ transport. High concentrations mimic the action of administered glucocorticoids in vivo, suggesting that a distinct pharmacological or toxic mechanism causes inhibition of Ca2+ absorption. This report further shows that dexamethasone (DEX) rapidly enhanced calbindin-D28K gene expression, that is de novo calbindin-D28K mRNA biosynthesis. DEX also markedly reduced the actions of RNA and protein synthesis inhibitors on calbindin-D28K gene expression, although no evidence for an action of DEX or 1,25-(OH)2D3 at the translational level was obtained. Ca2+ transport activity was highly correlated with calbindin-D28K concentration regardless of treatment. Washout permitted complete reversal of inhibition, verifying the specificity of inhibitor activity. These results appear to show positive contranscriptional regulation of calbindin-D28K gene expression by 1,25-(OH)2D3 and glucocorticoids. The use of this model should continue to clarify the interactive roles of nuclear-acting hormones on the Ca2+ absorptive mechanism and on complex physiological and pathological processes in general.

Differential effects of luteinizing hormone on intracellular free Ca2+ in small and large bovine luteal cells.

The effect of LH on the intracellular free Ca2+ concentration ([Ca2+]i) was investigated in highly purified small and large bovine luteal cell populations. Luteal cells were obtained from midcycle corpora lutea dispersed with collagenase and separated by flow cytometry into large and small cells. Resting levels of Ca2+ were higher (P less than 0.05) in the large than small cells [314 +/- 25 nM (n = 5) vs. 186 +/- 13 nM (mean +/- SE; n = 13) for large and small cells, respectively]. LH rapidly increased [Ca2+]i in both small and large cells loaded with the fluorescent Ca2+ probe fura-2. In the small cells, [Ca2+]i was immediately increased 2- to 6-fold (from 176 +/- 8 to 468 +/- 8 nM; n = 5) after adding LH. The LH induced [Ca2+]i rise occurred in two phases: an initial peak due to intracellular Ca2+ mobilization and a secondary rise due to Ca2+ influx from extracellular sources. Preincubation of the small cells with EGTA reduced the initial phase and abolished the secondary rise in [Ca2+]. Both forskolin and 8-bromo-cAMP increased [Ca2+]i in the small cells. In contrast, only a single phase of [Ca2+]i rise was observed in LH-treated large cells, and the response was 1.5- to 2-fold greater than the resting Ca2+ levels [314 +/- 25 vs. 435 +/- 60 nM (n = 4), for resting vs. LH-treated values, respectively]. The addition of both LH and prostaglandin F2 alpha (PGF2 alpha) to the large cells resulted in increases in [Ca2+]i that were greater than those induced by each hormone separately (2.0-fold for LH and 2.7-fold for PGF2 alpha vs. 7- to 9-fold in the presence of both hormones). These findings demonstrate that LH induces rapid increases in intracellular [Ca2+]i that differ in magnitude and profile between the small and large bovine luteal cells. Furthermore, LH and PGF2 alpha interacted to promote increases in [Ca2+]i in the large cells, that were higher than the sum of [Ca2+]i induced by each hormone separately.

Tissue-specific regulation of shell gland calbindin D28K biosynthesis by estradiol in precociously matured, vitamin D-depleted chicks.

Provision of Ca2+ for egg shell calcification in the avian uterus [egg shell gland (ESG)] derives mostly from vitamin D-dependent intestinal Ca2+ absorption from the diet. Ca2+ absorption is strongly linked to the intestinal vitamin D-dependent calbindin D28K (D28K) concentration. The laying hen ESG also contains D28K, and again, Ca2+ transport into the shell appeared to be linked to the ESG D28K concentration. However, evidence is now presented that ESG D28K synthesis may be estradiol (E2) dependent and vitamin D independent under certain conditions. One-day-old female chicks fed a vitamin D-free diet for as long as 6 weeks and then repeatedly injected im with E2 for up to 3 more weeks developed frank rickets, but possessed precociously matured reproductive tracts. While the tiny presumptive ESGs of nonestrogenized vitamin D-depleted chicks were devoid of D28K, the highly developed ESG, including the isthmus, of estrogenized chicks contained D28K. The ESGs of nonestrogenized, vitamin D-replete chicks also exhibited no development or detectable D28K. Regardless of whether vitamin D depleted or replete, estrogenized chick ESG contained similar D28K and D28K mRNA concentrations. Immunohistochemical techniques showed that the endometrial cellular localization of both D28K and Ca(2+)-ATPase (Ca2+ pump) in estrogenized chicks was similar to that in mature laying hens. There was no trace of D28K, nor was there any stimulation of Ca2+ absorption, in duodenum of vitamin D-free, immature chicks regardless of E2 treatment. As expected, both D28K and D28K mRNA were present in vitamin D-replete chick duodenum. We conclude that in E2-treated chicks, ESG D28K gene expression may be vitamin D independent and E2 dependent. This is the first clear demonstration of hormone-dependent tissue-specific D28K gene expression in the chick.

Synthesis and biological activity of novel vitamin D analogues: 24,24-difluoro-25-hydroxy-26,27-dimethylvitamin D3 and 24,24-difluoro-1 alpha,25-dihydroxy-26,27-dimethylvitamin D3.

We synthesized 24,24-difluoro-25-hydroxy-26,27-dimethylvitamin D3 (16), and 24,24-difluoro-1 alpha, 25-dihydroxy-26,27-dimethylvitamin D3 (21), from 3 beta-hydroxy-22,23-dinorcholenic acid 3-acetate. Compound 16 was found to be a highly potent vitamin D analogue with bioactivity similar to that of 25-hydroxyvitamin D3 in vivo. Compound 16 was bound by vitamin D binding protein with an affinity slightly less than that of 25-hydroxyvitamin D3. It was bound to the intestinal cytosol receptor for 1,25-dihydroxyvitamin D3 with approximately the same affinity as that of 25-hydroxyvitamin D3. In the organ-culture duodenum, 16 induced the synthesis of calcium binding protein with a potency approximately 1/20 that of 1,25-dihydroxyvitamin D3. Compound 21 was also noted to be a highly potent vitamin D analogue with bioactivity in vivo similar to that of 1,25-dihydroxyvitamin D3. It was bound to vitamin D binding protein with an affinity considerably less than that of 1,25-dihydroxyvitamin D3. It was bound to the intestinal cytosol receptor for 1,25-dihydroxyvitamin D3 with an affinity slightly less than that of the native hormone. In the organ-culture duodenum, 21 was noted to be about 3 times more active than 1,25-dihydroxyvitamin D3 in the induction of calcium binding protein. The introduction of fluorines at C-24 and extension of the sterol side chain at C-26 and C-27 by methylene groups results in vitamin D analogues that have biological activity in vivo similar to those of the respective nonfluorinated natural sterols.

1,25-Dihydroxyvitamin D3-induced calcium-binding protein: localization in organ-cultured embryonic chick duodenum.

1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) induces de novo biosynthesis of a specific calcium-binding protein (CaBP) in embryonic chick duodenum in organ culture. Using a highly sensitive and specific, peroxidase-antiperoxidase immunocytochemical procedure, 1,25(OH)2D3-induced CaBP in the organ-cultured duodenum was found only in the cytoplasm of absorptive cells, corresponding to its localization in rachitic chick duodenal cells after a single injection of 1,25(OH)2D3 in vivo. This observation, along with evidence correlating CaBP with calcium transport, strongly supports the use of the embryonic chick duodenal organ culture system as a physiologically relevant model of the vitamin D-dependent calcium absorptive mechanism.

Effects of verapamil and dexamethasone on the 1,25-dihydroxyvitamin D3-mediated calcium absorptive mechanism in the organ-cultured embryonic chick duodenum.

1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) is known to induce the biosynthesis of a specific, calcium-binding protein (CaBP) and to stimulate calcium transport in the organ-cultured embryonic chick duodenum. The biosynthesis of CaBP has been shown previously to exhibit an absolute dependence on the ambient calcium concentration of the culture medium. Verapamil, a calcium-channel blocker, decreased calcium influx into the organ-cultured duodenum and inhibited the induction of CaBP by 1,25(OH)2D3. Raising ambient calcium concentrations to as high as 10 mM did not prevent or reverse the inhibitory actions of verapamil. Dexamethasone, known to augment CaBP biosynthesis and calcium uptake in the organ-cultured duodenum in response to 1,25(OH)2D3, largely prevented inhibition of CaBP by verapamil. The actions of verapamil and dexamethasone were correlated with altered steady-state calcium concentrations of the organ-culture duodenum, strongly supporting a regulatory role of calcium in the 1,25(OH)2D3-mediated, intestinal calcium absorptive mechanism.

Calcitriol-dependent, paracellular sodium transport in the embryonic chick intestine.

1,25-Dihydroxyvitamin D3 (calcitriol), or vitamin D3 itself, when added to cultures of 20-day-old embryonic chick small intestine, stimulated sodium (Na+) uptake from the mucosal surface. The calcitriol-mediated increase in Na+ uptake appeared to be related to increased tight-junctional or paracellular permeability. Support for this conclusion was, first, that the uptake of other ions, potassium (K+) and rubidium (Rb+), with tight-junctional permeabilities greater than Na+, was also stimulated by calcitriol, and second, perturbation of cellular Na+ and K+ fluxes by inhibition of Na+/K+-ATPase activity did not affect calcitriol-stimulated Na+, K+, or Rb+ transport. Calcitriol stimulation of Na+ fluxes across the brush border as an alternate possibility is unlikely for the following reason: the calcium ionophore A23187, while mimicking the stimulatory action of calcitriol on calcium (Ca2+) uptake, reduced epithelial Na+ uptake. It is therefore suggested that calcitriol, by virtue of its effect on Ca2+ transport, reduces rather than stimulates cellular Na+ uptake.

Acute effects of prostaglandin F2 alpha on inositol phospholipid hydrolysis in the large and small cells of the bovine corpus luteum.

The present studies were conducted to determine whether the large or small bovine luteal cell was the site for the stimulatory effect of prostaglandin F2 alpha (PGF) on phospholipase C-catalyzed inositol phospholipid hydrolysis. Corpora lutea were removed from heifers during the luteal phase of the normal estrous cycle. Small luteal cells were isolated by unit-gravity sedimentation and large luteal cells were isolated by flow cytometry using a Becton Dickson FACS 440 cell sorter. PGF provoked rapid (5-30 s) and sustained (up to 30 min) increases in the levels of inositol mono-, bis-, and trisphosphates (IP, IP2, IP3, respectively) in small luteal cells. IP3 was formed more rapidly than IP2 or IP following PGF treatment. The PGF-stimulated increase in IP3 was accompanied by a transient reduction in the levels of 3H-labeled phosphatidylinositol 4,5-bisphosphate. LiCl (10 mM) enhanced inositol phosphate accumulation in response to PGF. Maximal increases in inositol phosphate accumulation were observed with 1-10 microM PGF and half-maximal increases were observed with 60 nM PGF. PGF (1-10 microM) had no effect on cAMP levels but stimulated small increases in progesterone accumulation in 30 min incubations of small luteal cells. PGF also increased the accumulation of inositol phosphates in large luteal cells. The increases were apparent within 5 min of incubation (the earliest time examined) and further increases were observed in incubations lasting 30 min. PGF had no significant effect on cAMP or progesterone in 30 min incubations of large cells.(ABSTRACT TRUNCATED AT 250 WORDS)

The synthesis and biological activity of 22-fluorovitamin D3: a new vitamin D analog.

We synthesized 22-fluorovitamin D3 from (22S) cholest-5-ene-3 beta, 22-diol-3 beta-acetate 2. Compound 2 was treated with diethylaminosulfur trifluoride to give 22-fluorocholest-5-en-3 beta-acetate 3 and (E) 22-dehydrocholest-5-en-3 beta-acetate. Compound 3 was treated with N-bromosuccinimide to give a mixture of the respective 5,7- and 4,6-dienes. The 5,7-diene of 3 was separated from the 4,6-diene using the dienophile 4-phenyl-1,2,4-triazoline-3, 5-dione. 22-Fluoro-5 alpha,8 alpha-(3,5-dioxo-4-phenyl-1, 2,4-triazolino)-cholest-6-en-3 beta-acetate 4 was purified by flash chromatography and treated with lithium aluminum hydride to generate 22-fluorocholesta-5,7-dien-3 beta-ol 5. Photolysis of the diene 5, followed by thermal equilibration, resulted in the synthesis of 22-fluorovitamin D3 7. The vitamin 7 increased active intestinal calcium transport only at a dose of 50,000 pmol/rat, whereas vitamin D3 increased intestinal calcium transport at a dose of between 50 and 500 pmol/rat. 22-Fluorovitamin D3 7 did not mobilize bone and soft tissue calcium at a dose as high as 50,000 pmol/rat, whereas vitamin D3 was successful in doing so at a dose of 500 pmol/rat. When tested in the duodenal organ culture system, 22-fluorovitamin D3 7 had approximately 1/25th the potency of vitamin D3. It did not antagonize the activity of 1,25-dihydroxyvitamin D3. 22-Fluorovitamin D3 7 bound to the rat plasma vitamin D binding protein less avidly than vitamin D3. 22-Fluorovitamin D3 was bound very poorly to the chick intestinal cytosol receptor for 1,25-dihydroxyvitamin D3. We conclude that the introduction of fluorine at the C-22 position results in a vitamin D sterol with decreased biologic activity when compared to vitamin D3. The presence of a fluorine group at C-22 position inhibits the binding of the vitamin to rat vitamin D binding protein when compared to the binding of its hydrogen analog, vitamin D3.

The effects of phenylbutazone on the intestinal mucosa of the horse: a morphological, ultrastructural and biochemical study.

Phenylbutazone, a non-steroidal anti-inflammatory drug known to produce intestinal erosions, was administered intravenously (13.46 mg/kg bodyweight) to 12 horses which were killed after 24, 48, 72 and 96 h. Eight untreated horses served as controls. Annular erosions in the duodenum and mucosal necrosis in the colon were seen after 48 h which progressed in severity. The erosions were characterised by sloughing of the surface epithelium, subepithelial cleft and bleb formation, necrosis of the lamina propria, degeneration of the walls of subsurface capillaries and microthrombosis. Large numbers of neutrophils with abundant fibrin and cellular debris were present at the erosion sites. Ultrastructurally, there was swelling of the endothelium of capillaries and small vessels, and of pericyte and smooth muscle cytoplasm in arterioles. In capillaries and post capillary venules, the endothelium ranged from swollen to lysed and necrotic. Extensive extravasation of erythrocytes and oedema were seen. These lesions were not seen in the control horses. Phenylbutazone produces a microvascular injury associated with the formation of duodenal and colonic erosions in horses. The duodenal and colonic mucosa were assayed at 48 and 96 h for prostacyclin and PGE2. There was no statistically significant difference between prostaglandin levels in the mucosa of control and treated horses. It was concluded that there was no correlation between mucosal prostaglandin levels and intestinal erosions after 48 h.

Ascorbate interacts with sodium selenite to increase glutathione peroxidase activity in selenium-deficient chick duodena cultured in vitro.

Duodena from Selenium (Se)/vitamin E-depleted 19 d chick embryos were cultured in vitro for 0-30 h. The addition of sodium selenite to the culture medium was associated with increased selenium-dependent glutathione peroxidase (SeGSHpx) activity after 24 h of incubation. In the absence of Se or in the presence of sodium ascorbate supplementation alone, SeGSHpx activity showed a gradual decline over the same time period. When ascorbate was added, along with sodium selenite, SeGSHpx activity was increased earlier and to a greater extent than in the presence of Se alone. These observations show that ascorbate can influence the metabolism of sodium selenite, resulting in increased SeGSHpx activity.

[A case of gastric lipoma. Diagnostic imaging]. / Presentazione di un caso di lipoma gastrico. Imaging diagnostico.

A case of gastric lipoma which manifested with an episode of acute gastrointestinal hemorrhage is reported. Preoperative diagnosis was based on the US, TC, and MRI findings, as the results of gastrointestinal endoscopy were inconclusive. The role of current imaging methods, and particularly of MRI, is discussed.

[Gallbladder carcinoma. Our experience]. / Carcinoma della colecisti. Nostra esperienza.

The authors report their experience of 18 patients with primary cancer of the gallbladder. On 10 patients at stage IV, 9 had a preoperative diagnosis, while at stage 0-1 and 2 the diagnosis was intraoperative or histologic. Every patient had a cholelithiasis at the same time. The authors discuss prophylactic cholecystectomy, even without specific symptoms, and emphasize the need for a better morphological and radiomorphological classification. In the light of the new microinvasive surgical techniques, they briefly discuss laparoscopic cholecystectomy and histologic diagnosis of carcinoma of the gallbladder.