High energy shock waves and 5-aminolevulinic for sonodynamic therapy: effects in a syngeneic model of colon cancer.

The cytotoxic effect of the natural porphyrin precursor 5-aminolevulinic acid (ALA) exposed to high energy shock waves (HESW) was investigated in vitro on DHD/K12/TRb rat colon cancer cells and in vivo on a syngeneic colon cancer model. In vitro, viable cell growth was determined by trypan blue exclusion assay and cell death was investigated by flow cytometry. ALA (50 µg/ml) and HESW (E1, EFD = 0.22 mJ/mm², 1000 shots or E2, EFD = 0.88 mJ/mm², 500 shots) showed a significant reduction of cancer cell proliferation at day 3 compared to cells exposed to ALA (p < 0.01) or HESW (p < 0.001) alone. An enhancement of necrotic and apoptotic cells was observed after combined treatment at day 1 with ALA and HESW E1 (a 3.1 and 6.4 fold increase vs ALA alone) or E2 (a 3.4 and 5.3 fold increase vs ALA alone). In vivo, apoptosis detection was carried out by TUNEL assay, the pro-apoptotic gene Bad and Bcl-2 mRNA expression was evaluated by quantitative SYBR Green real time RT-PCR and cleavage of poly(ADP-ribose)-polymerase (PARP) was investigated by Western Blotting. An enhancement of apoptosis was observed in tumour tissues after the combined treatment at day 1 with ALA (375 mg/kg i.v.) and HESW (E2) compared to that of ALA exposure alone with improved apoptotic index (a 2.0 fold increase), Bad enhanced mRNA expression (p < 0.01), Bcl-2 decreased mRNA expression (p < 0.05) and increased PARP cleavage. The interaction between HESW and ALA is then effective in inducing apoptosis on a syngeneic colon cancer model.

[The new hospital model established at the end of 19th Century and the professional nursing schools]. / La nascita del nuovo modello ospedaliero a fine Ottocento e la formazione professionale degli infermieri.

The historical research in hospitals organisation began in Europe only after the II World War. In particular, studies regarding the nursing in Italy are very scarce and heavily influenced by the fragmented political and organisational management due to the historical divisions within Italy. All the same, the social and working conditions, at least in the north of Italy, were similar in this respect. The workforce up to the beginning of the 20th Century was equally divided between males and females. The female recruitment took place exclusively from the foundlings left in front of the hospital and raised within the hospital walls. The work was very hard with long working hours, under a total subordination to the religious administrators with miserable wage. These conditions remained unchanged until the second half of the 19th Century when, in central Europe, a change in the reorganisation of hospital management occurred, together with a cultural improvement in the nursing profession. This process of reforms had started in England some decades earlier from the reorganisation of nursing made by Florence Nightingale. After Italian post-unification the healthcare field was involved in the tentative to homologate the vast differences in the hospital organisation within the Italian states which also generated tension, bitterness and many political debates. The particular conditions created in the main hospital of Turin in the second half of the 19th Century, together with the political and social climate at the time and the reopening of debates between Socialist, Liberals and Catholics favoured the reorganizational improvement of the San Giovanni Battista hospital. Under this prospective, the foundation of the nursing school came into being, preceded by the intense control of a behavioral and ethical cleaning of the nurses, in particular the males. The availability of the hospital administrations in carrying out the recommendations of the medical staff in the modernisation and sanitary improvements and also in the quality of service was manifested by the decision to keep only the personnel who had obtained qualifications at the ''Nursing School''. This was also due to the financial support of the school itself. A further indication of appreciation for the school activity was the premature decision to extend the course to two years. This formative activity rapidly showed positive results, improving service quality and raising the professional ethics of the nurses. A few years after the foundation of the Nursing School in Turin, the school was contacted by numerous Italian hospital administrations to obtain information on the practices and organisational activities of the school. Thus, the process of professional training of nursing in Italy gradually started to take hold. At the same time, despite the diffusion of the hospital nursing schools, the experience of the Nightingale style of schooling in Italy never firmly established with success. The continuity of the professional instruction is instead to be noted in the transformation of the hospital boarding schools, exclusively for females, which in turn evolved into the classic Italian professional nursing school.

Sex steroid receptors, secondary bile acids and colorectal cancer. A possible mechanism of interaction.

AIM: The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics, as sex steroid receptors. METHODS: Specific androgen (AR), estrogen (ER) and progesterone (PgR) receptors were measured in the tissue samples of 35 patients (15 males, 20 females) undergoing colectomy or coloproctectomy for adenocarcinoma. The characteristics of androgen receptor (AR, DHT-R: dihydrotestosterone receptor) were also investigated using competitive activity of cyproterone acetate, cortisol, aldosterone and steroid-like substances such as deoxycholic and lithocholic acid, present in the milieu of the considered organ. Binding assays and competition tests were conducted using a charcoal dextran method. RESULTS: When present (50%), ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa. AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa, with no significant difference. Androgen receptor however exhibited an altered binding activity in cancer specimens. Cyproterone acetate did not displace DHT from AR while significant displacing activity was elicited by DHT, testosterone, as well as by lithocholic acid, but not by deoxycholic acid. CONCLUSION: In cancerous large bowel mucosa, androgen receptors show altered binding characteristics. The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event.

Transmucosal transport of tobramycin incorporated in solid lipid nanoparticles (SLN) after duodenal administration to rats. Part II--tissue distribution.

Tobramycin-loaded solid lipid nanoparticles (SLN) were prepared and administered by duodenal and intravenous (i.v.) routes to rats and the tissue distributions were determined successively at fixed times (30 min, 4 h and 24 h) and compared to those of the tobramycin solution after i.v. administration. The tissue distribution between tobramycin-loaded SLN administered duodenally and i.v. was different. A marked difference between tobramycin-loaded SLN administered duodenally and tobramycin solution administered i.v. was also evidenced. In particular, the amounts of tobramycin in the kidneys after tobramycin-loaded SLN administration either duodenally or i.v. were lower than after administration of i.v. solution. Tobramycin-loaded SLN were able to pass across the blood-brain barrier in rats to a greater extent after i.v. injection than after duodenal administration.

Transmucosal transport of tobramycin incorporated in SLN after duodenal administration to rats. Part I--a pharmacokinetic study.

Tobramycin-loaded solid lipid nanospheres (SLN) were prepared and administered to rats into the duodenum; their behaviour was compared to that of tobramycin-loaded SLN administered intravenously (i.v.). A tobramycin control solution was also administered to rats. Tobramycin in solution is not absorbed by the gastrointestinal route, while tobramycin incorporated in the SLN is absorbed. A high concentration of tobramycin is still present in plasma 24 hours after the duodenal administration of tobramycin-loaded SLN. Tobramycin-loaded SLN administered i.v. showed a prolonged circulation time compared to the i.v. administered tobramycin solution. The AUC of tobramycin in SLN administered duodenally is higher than those of tobramycin in SLN and in solution administered i.v.

Non-stealth and stealth solid lipid nanoparticles (SLN) carrying doxorubicin: pharmacokinetics and tissue distribution after i.v. administration to rats.

Non-stealth and stealth solid lipid nanoparticles (SLN) carrying doxorubicin were prepared as drug delivery systems. The pharmacokinetics and tissue distribution of doxorubicin in these SLN were studied after i.v. administration to conscious rats and were compared to the commercial solution of doxorubicin. The same dose of each formulation (6 mg kg(-1)of body weight) of doxorubicin was injected in the rat jugular vein. Blood samples were collected after 1, 15, 30, 45, 60 min and 2, 3, 6, 12, and 24 h after the injection. Rats were sacrificed after intervals of 30 min, 4 h, and 24 h and samples of liver, spleen, heart, lung, kidney, and brain were collected. In all samples, the concentration of doxorubicin and of the metabolite, doxorubicinol, were determined. Doxorubicin and doxorubicinol were still present in the blood 24 h after injection of stealth and non-stealth SLN, while they were not detectable after the injection of the commercial solution. The results confirmed the prolonged circulation time of the SLN compared to the doxorubicin solution. In all rat tissues, except the brain, the amount of doxorubicin was always lower after the injection of the two types of SLN than after the injection of the commercial solution. In particular, SLN significantly decreased the heart concentration of doxorubicin.

Pharmacokinetics of doxorubicin incorporated in solid lipid nanospheres (SLN).

The pharmacokinetics of doxorubicin incorporated as ion-pair into solid lipid nanospheres (SLN) was compared with that of the commercial solution of the drug. Male albino rats (Wistar-derived strain) were treated i.v. with equivalent doses (6 mg kg(-1)) of two different doxorubicin formulations: an aqueous dispersion of SLN carrying doxorubicin and a commercial doxorubicin solution (Adriablastina). These formulations were injected, under general anaesthesia, through a cannula into the jugular vein and blood samples were collected at 1, 15, 30, 45, 60, 120 and 180 min after administration. After 180 min rats were killed and samples of liver, heart, lung, kidney, spleen and brain were collected. Blood and tissue samples were analysed by a spectrofluorimetric method. The anthracycline concentration in the blood was markedly higher at each point times with the SLN than with the commercial solution. The drug concentration was also higher in the lung, spleen and brain. SLN-treated rats showed a lower doxorubicin concentration in liver, heart and kidney. The results showed that SLN increased the area under the curve (0-180 min) of doxorubicin compared to conventional doxorubicin solution and led to a different body distribution profile.

Solid lipid nanoparticles in lymph and plasma after duodenal administration to rats.

PURPOSE: To evaluate the uptake and transport of solid lipid nanoparticles (SLN), which have been proposed as alternative drug carriers, into the lymph and blood after duodenal administration in rats. METHODS: Single doses of two different concentrations of aqueous dispersions of unlabelled and labelled SLN (average diameter 80 nm) were administered intraduodenally to rats. At different times, samples of lymph were withdrawn by cannulating the thoracic duct and blood was sampled from the jugular vein. Monitoring continued for 45 and 180 minutes, for unlabelled and labelled SLN respectively. The biological samples were analysed by photon correlation spectroscopy (PCS), transmission electron microscopy (TEM) and gamma-counting. RESULTS: TEM analysis evidenced SLN in lymph and blood after duodenal administration to rats: the size of SLN in lymph did not change markedly compared to that before administration. The labelled SLN confirmed the presence of SLN in lymph and blood. CONCLUSIONS: The uptake and transport of SLN in the lymph, and to a lesser extent in the blood, were evidenced. The in vivo physical stability of SLN may have important implications in designing drug-carrying SLN.

Primary hyperaldosteronism. Analysis of risk factors associated with persistent postoperative hypertension.

Between 1983 and 1992, 24 patients with primary hyperaldosteronism were operated at our division. Among these, 21 had adrenal adenoma and constituted the subjects of our study. The aim of this retrospective study was to determine risk factors that could potentially lead to persistent arterial hypertension (5 cases or 23.9%). The variables of the univariate and the multivariate analyses were: sex, age, diameter of the neoplasm, preoperative duration of the hypertension, and familial hypertension. The analysis demonstrated no statistically significant difference between the resolution of hypertension and the variables. Though not statistically significant, the only variable that approached statistical significance (p < 0.11) was familial hypertension.

Modifications of the trophism of intestinal mucosa after intestinal and bilio-pancreatic diversion in the rat.

Morphofunctional alterations to the intestinal mucosa are influenced by three main factors: food, bilio-pancreatic secretions, intestinal hormones. In order to assess the importance of each one, histological and histochemical tests were performed on different segments of intestine taken from rats which were sacrificed one month after the following procedure: gastrojejunal anastomosis on a Roux loop (Model I); same procedure plus biliopancreatic bypass into the jejunum (Model II). When compared to the controls, Model I duodenum samples revealed hypertrophy of the entire wall with "bundles" of villi, while Model II samples showed a clear hypotrophy and reduction in the number of duodenal villi. To such modifications of the duodenum correspond longer, thinner villi in the other bowel segments, particularly in the jejunum and distal ileum. These results suggest that the predominant trophic effect derives from contact with biliopancreatic secretions at a proximal level. The modifications of the duodenal mucosa appear to regulate the trophism of the distal segments probably by the secretion of distant acting enterohormones.

[Suspensory elements of the anal canal. Anatomo-surgical considerations]. / Elementi di sospensione del canale anale. Considerazioni anatomo-chirurgiche.

Continence depends not only on correct muscular function but also on the integrity of the suspensory structures of the anal canal, the latter being the subject of this study performed on embryological and adult specimens. The role of the perineal body as an insertion tendon common to all the striated muscles (pubo-coc-cygeus, superficial striated sphincter) which anchor the anal canal to the surrounding structures is pointed out. The characteristic behaviour of the longitudinal muscle that links the striated musculature to the visceral cylinder is also described.

Gluten-induced experimental IgA glomerulopathy.

The effect of alimentary gluten and of its lectin-like fraction gliadin in inducing IgA mesangial deposits in BALB/c mice was investigated. In the pilot study G1 (4 mice), G2 (4 mice) and G3 (6 mice) received ovalbumin, human gamma-globulins, and crude gluten, respectively. The antigens, 1 mg/ml, were given in drinking water for 14 weeks. G4 (20 mice) were fed with standard mouse fodder. Gluten, as well as other alimentary antigens, induced IgA mesangial deposits with intense IgA staining in each animal, however a positive IgA staining was also observed in 10 of 20 adult controls. Antigliadin IgA antibodies were detected in renal tissue eluates from gluten-immunized mice but were found in eluates from control animals too. IgA deposits and antigliadin IgA in renal tissue were also observed in 3 of 7 adult mice (G7) fed for 30 weeks with standard fodder, then for 1 month with a protein-free diet supplemented with 20% amino acids. Conversely, there were no IgA mesangial deposits or IgA anti-gliadin antibodies in renal eluates of 1- or 2-week-old mice (G5 and G6). For the definitive protocol, 4-week-old BALB/c mice were selected and fed with basal glutenfree diet. G8 (14 mice) did not receive any alimentary immunogen in the drinking water, whereas G9 (15 mice) and G10 (15 mice) received ovalbumin and gliadin, respectively. G11 (15 mice) had standard gluten-containing diet. IgA deposits semiquantified by immunofluorescence scores were found to be significantly greater in G9 and G10 than in G8 (Student' t-test p1 less than 0.003, Mann-Whitney test p2 less than 0.001 and p1 less than 0.01, p2 less than 0.007, respectively), and in G11 than in G8 (p1 and p2 less than 0.05). The presence of positive IgA staining (greater than or equal to 2/6 scores) was significantly less frequent in G8 in comparison to G9 (chi-square test p3 less than 0.002), G10 (p3 less than 0.02) and G11 (p3 less than 0.04). Total serum IgA were significantly higher in orally immunized G9 and G10 than in G8 control mice (p1 less than 0.005, p2 less than 0.002). Anti-gliadin IgA in circulation as well as in renal deposit eluates were significantly increased in gluten-eating mice (G10 and G11) as compared with the gluten-free control group G8. These observations indicate that gliadin does induce IgA immune deposits in BALB/c mice.(ABSTRACT TRUNCATED AT 250 WORDS)

[Wirsung-septoplasty during interventions on Vater's ampulla]. / La Wirsung-settoplastica in corso di interventi sulla papilla di Vater.

After a short survey of pathogenetic hypotheses for pancreatitis during papillary lithiasis, stress is laid on the importance of the use of the microscope in peroperative diagnosis of infundibulo-papillar pathology and in the surgery of distal Wirsung.

[Role of the total and prolonged diversion of portal blood on hepatic function in the rat]. / Ruolo della derivazione totale e protratta del sangue portale sulla funzionalita' epatica nel ratto.

In this experience the effect on hepatic parenchyma and function has been analyzed 180 days after total portacaval shunt. The portacaval shunt induces 50% mortality rate at six months. In the survived animals a significant reduction of hepatic mass has been observed. The liver still presents significant elevation of necrosis and cholestasis enzymes. The residual hepatocellular function related to hepatic mass is comparable to sham operated rats, but it is quite insufficient to maintain endogenous clearances.

[Intestinal adaptive response of the rat after bilio-pancreatic and food diversion from the duodenum]. / La risposta adattativa intestinale del ratto dopo diversione bilio-pancreatica ed alimentare dal duodeno.

The object of the research was to evalute, in the rat, the effects brought about on the intestinal trophism by the selective and combined exclusion of the bile, the pancreatic secretion and the food exsert, certainly, a trophic activity on the intestinal mucous membrane where they pass trough, as it has been observed by other authors; but the modifications, observed on last segments of the ileum, do not explain a direct action of these factors to modulate the adaptative intestinal reaction. In the following essay we propose the hypothesis that the privation of biliary and pancreatic secretions and the food from the duodenum may stimulate a hormonal and duodenal reaction which is able to produced observed phenomena.