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The anticoccidial activity of thymol, carvacrol, and saponins was assessed in an in vitro model of coccidiosis. Eimeria spp. sporozoites were collected from field samples, characterized and used for 2 different invasion assays on Madin-Darby Bovine Kidney cells (MDBK). The cells were challenged with 5 x 104 sporozoites without (control) or with various treatments: saponins (10 ppm), thymol and carvacrol (7 ppm each), or a combination of saponins, thymol, and carvacrol at 2 doses; MIX 1 (saponins 5 ppm, thymol 3.5 ppm and carvacrol 3.5 ppm) and MIX 2 (saponins 10 ppm, thymol 7 ppm and carvacrol 7 ppm). The treated cells were incubated at 37°C for 24 hours (invasion assay 1) and for 2, 24, and 48 hours (invasion assay 2). The efficiency of invasion was determined by counting the sporozoites left in the supernatant that were not able to invade the cells, while intracellular Eimeria DNA was detected by qPCR to confirm the data. Data were analyzed with ANOVA and differences were considered significant when p-value was ≤ 0.05. Data from invasion assay 1 showed that the thymol and carvacrol-containing blends significantly reduced invasion, especially in combination with saponins at the highest dose. Saponins alone did not have a strong inhibiting activity but acted synergistically with the other molecules. Interestingly, in invasion assay 2 it was found that the effect of the highest dose of the blend of saponins, thymol and carvacrol was already visible at 2 hours post infection, while the other treatments were significantly successful at 24 hours post infection. The invasion assay protocol was designed to screen molecules in vitro starting from field fecal samples and it can respresent a potential tool in Eimeria research. Moreover this study shows that invasion in MDBK cells by Eimeria sporozoites is inhibited in presence of thymol, carvacrol and saponins, thus highlighting the anticoccidial potential of these compounds.

We recently described the genetic antimicrobial resistance and virulence profile of a collection of 279 commensal E. coli of food-producing animal (FPA), pet, wildlife and human origin. Phenotypic antimicrobial resistance (AMR) and the role of commensal E. coli as reservoir of extra-intestinal pathogenic Escherichia coli (ExPEC) virulence-associated genes (VAGs) or as potential ExPEC pathogens were evaluated. The most common phenotypic resistance was to tetracycline (76/279, 27.24%), sulfamethoxazole/trimethoprim (73/279, 26.16%), streptomycin and sulfisoxazole (71/279, 25.45% both) among the overall collection. Poultry and rabbit were the sources mostly associated to AMR, with a significant resistance rate (p &gt, 0.01) to quinolones, streptomycin, sulphonamides, tetracycline and, only for poultry, to ampicillin and chloramphenicol. Finally, rabbit was the source mostly associated to colistin resistance. Different pandemic (ST69/69*, ST95, ST131) and emerging (ST10/ST10*, ST23, ST58, ST117, ST405, ST648) ExPEC sequence types (STs) were identified among the collection, especially in poultry source. Both ST groups carried high number of ExPEC VAGs (pandemic ExPEC STs, mean = 8.92, emerging ExPEC STs, mean = 6.43) and showed phenotypic resistance to different antimicrobials (pandemic ExPEC STs, mean = 2.23, emerging ExPEC STs, mean = 2.43), suggesting their role as potential ExPEC pathogens. Variable phenotypic resistance and ExPEC VAG distribution was also observed in uncommon ExPEC lineages, suggesting commensal flora as a potential reservoir of virulence (mean = 3.80) and antimicrobial resistance (mean = 1.69) determinants.

Background Enterocytes exert an absorptive and protective function in the intestine, and they encounter many different challenging factors such as feed, bacteria, and parasites. An intestinal epithelial in vitro model can help to understand how enterocytes are affected by these factors and contribute to the development of strategies against pathogens. Results The present study describes a novel method to culture and maintain primary chicken enterocytes and their characterization by immunofluorescence and biomolecular approaches. Starting from 19-day-old chicken embryos it was possible to isolate viable intestinal cell aggregates that can expand and produce a self-maintaining intestinal epithelial cell population that survives until 12 days in culture. These cells resulted positive in immunofluorescence to Cytokeratin 18, Zonula occludens 1, Villin, and Occludin that are common intestinal epithelial markers, and negative to Vimentin that is expressed by endothelial cells. Cells were cultured also on Transwell® permeable supports and trans-epithelial electrical resistance, was measured. This value gradually increased reaching 64 Ω*cm2 7 days after seeding and it remained stable until day 12. Conclusions Based on these results it was confirmed that it is possible to isolate and maintain chicken intestinal epithelial cells in culture and that they can be suitable as in vitro intestinal model for further studies.

Formaggio di Fossa di Sogliano is a traditional Italian Protected Designation of Origin (PDO) cheese ripened for a minimum of 5 months, with the feature of a ripening of at least 80 to at most 100 days in pits, digged into tuffaceous rocks according to medieval tradition of Italy. In this study, a challenge test using Listeria innocua as a surrogate of Listeria monocytogenes was performed, with the aim of increasing knowledge concerning the impact of the Fossa cheese process, and especially of the traditional ripening process of this PDO, on the behaviour of L. monocytogenes. Pasteurized milk was experimentally inoculated with 4.5 log CFU/mL cocktail by three L. innocua strains, and L. innocua and Mesophilic Lactic Acid Bacteria (LAB) counts as well as the evolution of temperatures, pH and aw values were monitored throughout the manufacturing and ripening processes. Throughout the ripening in maturation room a constant temperature of 8°C was observed reaching a temperature between 10 and 15.5°C during ripening into pit. In the final products data for LAB concentration, pH and aw values were roughly in accordance with literature, even if some differences were, probably due to variability of artisanal cheese productions. The numbers of L. innocua showed a slight decrease but remained stable until the end of ripening in maturation room, whereas a significant reduction of the microorganism was observed in the final product, at the end of the ripening into the pit. The findings give scientific evidence that the process of this PDO prevented the L. innocua growth, allowing us to speculate a similar behaviour of L. monocytogenes. Based on this study, the recommendation to extend as much as possible the ripening into pit (from 80 to 100 days) was provided to food business operators as a risk mitigation strategy to be implemented.

Salmonella spp. represent a public health concern for humans and animals due to the increase of antibiotic resistances. In this scenario, the use of essential oils (EOs) could be a valid tool against Salmonella contamination of meat. This work compares the in vitro effectiveness of an Italian mixture of feed additives based on EOs (GR-OLI) with EO of Origanum vulgare L., recently admitted by European Food Safety Authority (EFSA) for animal use. Twenty-nine Salmonella serotypes isolated from poultry and pig farms were used to assess GR-OLI and O. vulgare EO antimicrobial propeties. O. vulgare EO was active on the disaggregation of mature biofilm, while GR-OLI was capable of inhibiting biofilm formation and disaggregating preformed biofilm. Furthermore, GR-OLI inhibited bacterial adhesion to Caco-2 cells in a dose-dependent manner. Both products showed inhibition of bacterial growth at all time points tested. Finally, the synergistic action of GR-OLI with commonly used antibiotics against resistant strains was investigated. In conclusion, the mixture could be used both to reduce the meat contamination of Salmonella spp. before slaughter, and in synergy with low doses of ciprofloxacin against resistant strains. Although EOs as feed additives are already used in animal husbandry, no scientific study has ever highlighted their real antimicrobial potential.

Chestnut tannins (CT) and saturated short medium chain fatty acids (SMCFA) are valid alternatives to contrast the growth of pathogens in poultry rearing, representing a valid alternative to antibiotics. However, the effect of their blends has never been tested. Two blends of CT extract and Sn1-monoglycerides of SMCFA (SN1) were tested in vitro against the proliferation of Clostridium perfringens, Salmonella typhymurium, Escherichia coli, Campylobacter jejuni. The tested concentrations were: 3.0 g/kg of CT, 3.0 g/kg of SN1, 2.0 g/kg of CT and 1.0 g/kg of SN1, 1.0 g/kg of CT and 2.0 g/kg of SN1. Furthermore, their effect on broiler performances and meat quality was evaluated in vivo: one-hundred Ross 308 male birds were fed a basal diet with no supplement (control group) or supplemented with CT or SN1 or their blends at the same concentration used in the in vitro trial. The in vitro assay confirmed the effectiveness of the CT and SN1 mixtures in reducing the growth of the tested bacteria while the in vivo trial showed that broiler performances, animal welfare and meat quality were not negatively affected by the blends, which could be a promising alternative in replacing antibiotics in poultry production.

Essential oils (EOs) have deserved much attention in the past decades for their antimicrobial activity, since many of them have demonstrated efficacy against food-borne pathogenic and spoilage microorganisms. Moreover, they have potential application in animal nutrition as multifunctional feed supplements, avoiding or diminishing the use of antibiotics in livestock. However, low solubility and bioavailability as well as volatility and marked aromatic note are important limitations in food and feed applications. In this study we present the synthesis, characterization and evaluation of the antibacterial activity of new thymol, carvacrol and menthol derivatives. The new compounds have been designed to overcome the limitations of the precursors, such as poor water solubility and volatility, still maintaining a good antimicrobial profile. We evaluated the activity of the synthetized compounds against pathogens causing important foodborne diseases, i . e . Clostridium perfringens , Salmonella typhimurium , Salmonella enteritidis and Escherichia coli . The low MICs and MBCs values for some of the studied compounds, combined with water solubility and negligible cytotoxicity towards HT-29 human cells, confirmed the potential use for EOs derivatives in the food industry.

Miocene postcollisional porphyry Cu deposits in southern Tibet are genetically associated with dacitic-rhyolitic intrusions with unusually high Sr/Y ratios (>40), which have been attributed to dehydration melting of garnet amphibolite in a thickened lower crust. To test this hypothesis and examine the hydration state of copper ore-forming high Sr/Y magmas, we utilize a geohygrometer for granitoid rocks, entailing zircon-saturation thermometry and H 2 O-dependent phase equilibria. The results show that these Tibetan high Sr/Y magmas had dissolved H 2 O contents >10 wt%, which considerably exceeds the water supply by dehydration melting of basaltic amphibolites (maximum of 6.7 ± 1.4 wt%). Our results indicate that high Sr/Y dacitic-rhyolitic magmas cannot be produced by dehydration melting of basaltic amphibolites. While H 2 O-added melting of basaltic amphibolites can produce high Sr/Y dacitic-rhyolitic melts, it does not yield high enough Mg# (>50) to match the Tibetan ore-forming porphyries. We propose an alternative model for the genesis of copper ore-forming high Sr/Y magmas in Tibet, and suggest that the high Sr/Y dacitic-rhyolitic porphyries in southern Tibet are residually H 2 O-enriched, high-pressure differentiation products of hydrous mafic partial melts of Tibetan mantle. This hypothesis is based on the previous investigation of Miocene mafic microgranular enclaves (mantle-derived melts), which define a fractionation trend with, and have Sr-Nd-Hf isotopic compositions similar to, the host Tibetan ore-forming porphyries.

Infectious bronchitis virus (IBV) represents one of the poultry industry major threats, particularly in high density producing countries. The emergence and spread of new IBV genotypes have frustrated the various disease control efforts implemented over time. Despite that, few comprehensive and large scale studies have been performed to understand the international and local spreading dynamics of this virus. In the present work, these phenomena were evaluated by implementing a Bayesian phylodynamic approach to reconstruct the epidemiological patterns and population history of the QX genotype (currently renamed GI-19 lineage), the most relevant IBV lineage of the Old-World. Our analysis, based on 807 partial S1 sequences of strains collected from 18 countries between 1993 and 2015, demonstrates that this genotype originated in China well before its first identification. After a prolonged local circulation, it started spreading to other European, Asian and Middle East countries in successive waves, which were mirrored by concomitant fluctuations in viral population size. Interestingly, the within-Europe spread was characterized by a higher estimated migration rate compared with the inter-continental one, potentially reflecting the closer geographic and economic relationships among these countries. Nevertheless, the colonization of new states by the GI-19 lineage appeared to occur mostly by single introduction events in both intra and inter-continental spread, likely because of epidemiological factor and health policy combination which seems to prevent the frequent introduction and mixing of different strains. On the other hand, the within Italy QX circulation reconstruction showed a much more intricate connection network among different locations, evidencing the difficulty in controlling IBV spread especially in highly densely poultry populated areas. The presence of several well supported epidemiological links among distantly related Italian regions testifies that animal transportation and indirect transmission routes rather than local airborne diffusion contribute to the QX success and persistence at local scale. Globally, the spreading dynamics and evolution of the QX genotype were reconstructed from its very origin to nowadays, demonstrating the need of more effective direct control measures, particularly within each country. Unfortunately, the incompleteness of available molecular epidemiology data represents an insurmountable limit which leaves many questions currently unsolved, thus highlighting the compulsoriness of a structured monitoring and data sharing system implementation.

Objective To investigate some pathogenic characters of Salmonella enterica strains isolated from poultry. Methods Twenty-three genetically distinct Salmonella enterica strains, of different serovars and pulsotype, were examined for virulence traits. Resistance to gastric acid environment was estimated by measuring the percentage of survived bacterial cells after exposure for 2 h to a synthetic gastric juice. Strains were analyzed with PCR for the presence of the following virulence genes: mgtC and rhuM located on SPI-3, sopB and pipB located on SPI-5, Salmonella virulence plasmid (spv) R (spvR), spvB and spvC located on Salmonella plasmid virulence and sodCI, sopE, and gipA located on prophage. Finally, resistance to 21 antibiotics was tested with Kirby–Bauer method. Results A percentage of 82.60% of strains were resistant to gastric environment after induction and 60.87% of the strains exhibited constitutive resistance too. Nineteen different virulence profiles were detected. The phage related genes sodCI and sopE and the plasmid mediated operon spvR, spvB and spvC (spvRBC) were detected in 82.60%, 47.82% and 52.17% of strains, respectively. Typhimurium and Enteritidis strains showed the highest number of virulence genes. Twenty-one different antibiotic resistance profiles were obtained and two isolates (Typhimurium and Enteritidis) resulted sensible to all the tested molecules. The ampicillin, streptomycin, sulfonamide and tetracycline resistance profile was detected in seven isolates (30.43%). Conclusion Our results show that paratyphoid Salmonella strains with several characters of pathogenicity, that may be cause of severe pathology in animals and humans, are circulating among poultry.

Workplaces have been recognized as favored context for health promotion as they directly affect workers’ physical, psychological and social well-being. The IRCCS Policlinico San Matteo Foundation, in Pavia, has great potential as a setting for health promotion due to its significant workforce, including approximately 3000 employees in 2015. This is therefore well-suited to address both occupational health issues and …

In April 2009, a novel H1N1 influenza A virus (pH1N1) was recognized as the cause of the flu pandemic in humans. Here, we report the isolation of pH1N1 virus from the lung homogenates of two cats, which died after severe respiratory symptoms. The cats belonged to a cat colony consisting of 90 caged cats and were found dead following a 2-week period of respiratory and gastrointestinal diseases in the colony. During the outbreak, 25 cats died and 50% of the animal colony showed anorexia, depression, respiratory and gastrointestinal symptoms. Histological examination of the lungs of the two tested cats displayed lesions centred on terminal airways with epithelial bronchiolar hyperplasia and alveolar necrosis. Influenza A virus was detected in the lung tissues by immunohistochemistry and real-time RT-PCR (rRT-PCR). Partial sequences of haemagglutinin (HA) genes and complete sequences of neuraminidase (NA) genes of the two isolates displayed high similarity to the pH1N1 viruses circulating in humans (99% for HA gene and 100% for NA gene). To determine whether the pandemic virus had circulated among cats, serum samples and pharyngeal swabs were collected from 38 cats of the colony. Serum samples were tested by ELISA to detect antibodies against pH1N1 nucleoprotein and by hemagglutination-inhibition test, while pharyngeal swabs were examined by pH1N1 specific rRT-PCR. Twenty-one (55%) of the tested cats carried antibodies against the isolated strain and two swabs were positive for the presence of pH1N1 RNA. Our results confirm that the pH1N1 virus was able to infect cats and raise the hypothesis of the circulation of the virus within the colony being due to cat-to-cat transmission. The case reported here provides, to the best of the authors' knowledge, the first description of the pH1N1 infection involving numerous cats that lived in a restricted area with limited contact with humans.

Palladium (Pd) is the most mobile element of the platinum group elements (PGE) in hydrothermal fluids. The characterization of the nature and stability of Pd(II) complexes in geofluids is essential in understanding the formation of hydrothermal PGE deposits and the remobilization of PGE during hydrothermal and metamorphic overprints of magmatic deposits. However, the aqueous speciation of this metal in a range of geologically relevant conditions remains controversial. A number of experimental studies of Pd solubility and speciation in hydrothermal fluids suggest that chloride and bisulfide are the major ligands responsible for carrying Pd as Pd(II)–Cl and Pd(II)–HS complexes, but different experimental studies predicted different predominant chloride and bisulfide complexes and their relative strengths. Hence, we conducted ab initio molecular dynamics (MD) simulations to predict the speciation of Pd–Cl and Pd–HS complexes at 300 °C. The simulations predicted that all complexes share fourfold square-planar structures, which is consistent with X-ray absorption spectroscopy measurements of Pd(II) in chloride-rich solutions. The stability constants for the stepwise formation of Pd(II)–Cl and Pd(II)–HS complexes were determined using thermodynamic integration. The predicted formation constants of Pd(II)–Cl complexes show excellent agreement (within ∼1 order of magnitude for PdCl + , within 0.3 for PdCl 2(aq) and PdCl 3 − , within 0.1 for PdCl 4 2− ) with the recent experimental study of Tagirov et al. (2013). However, our results suggest that the Pd(HS) 4 2− complex predominates in HS − -rich hydrothermal fluids, whereas interpretation of previous experimental studies neglected this species. Modeling of Pd solubility in chloride- and sulfur-rich hydrothermal fluids demonstrated that Pd is mainly carried as the Pd(HS) 4 2− hydrosulfide complex at neutral-alkaline and reduced (pyrite/pyrrhotite stable) conditions, and as the PdCl 4 2− chloride complex at acidic and oxidized conditions. At 300 °C, significant Pd mobility at ppb level as Pd bisulfide complexes is predicted under fluid-buffered conditions (e.g., pH ∼7 to 8, near HS − /H 2 S(aq) pH buffer), but only limited Pd solubility is predicted under rock-buffered conditions (e.g., pH ∼4 to 5, quartz–feldspar–muscovite buffer).

Lower tract involvement of systemic sclerosis is an uncommon manifestation. In this study we have evaluated the functional and morphologic vesical changes in 23 female patients with systemic sclerosis to investigate if bladder dysfunction was correlated with an extended skin sclerosis and/or a more serious organ involvement. Urodynamic alterations were observed in only 3 cases. Urinary symptoms and urodynamic features were correlated neither with severity of vesical fibrosis nor with visceral involvement. Autonomic dysfunction was detected in 13 patients.

Endometriosis of the urinary tract is a relatively rare condition. Since clinical signs are not specific the diagnosis is difficult and the therapy is not well defined. Two cases and a review of the literature are presented.

The use of absorbable staples may allow to reduce the operating time of orthotopic bladder replacement. We report our experience in 14, patients with a modification of the Studer technique using absorbable PolyGIA instruments. The technique has been shown to be simple and quick to perform (time for pouch creation 15–40 minutes, mean 16 minutes) with no significant intraoperative difficulties. Urodynamic data and continence are satisfactory and seem comparable to different procedures and not related to staples' use. The main question remains if the reduced operating time equalizes the high cost of staple devices.

Proximate composition and cholesterol content, fatty acid and amino acid profiles, selected mineral (Na, K, Mg, Ca, P, Fe, Zn) and vitamin (niacin, pantothenic acid, pyridoxine, vitamin B12) content, total and soluble collagen content, and purine bases (adenine, guanine, xanthine, hypoxanthine) were determined in cultured sturgeon. White sturgeon (Acipenser transmontanus), Italian sturgeon (Acipenser naccarii) and Siberian sturgeon (Acipenser baeri) were individually analyzed. The results were reported as a whole, since no commercial distinction is made between the three species. Mean content (per 100 g wet weight) of protein, lipid, ash, and cholesterol was 19.23 g, 7.63 g, 1.09 g, and 66 mg, respectively. Saturated, monounsaturated, and polyunsaturated (PUFA) fatty acid contents were 1.76, 3.12, and 1.46 g/100 g wet weight, respectively;n− 3 PUFAs reached 1.18 g/100 g, whereasn− 6 PUFAs were 0.28 g/100 g. Then− 3/n− 6 ratio was 4.23. Compared to human requirements, sturgeon protein was especially rich in histidine and isoleucine (0.82 and 0.92 g/100 g wet weight, respectively) but rather poor in tryptophan (0.14 g). The Mg concentration was fairly good (46.8 mg/100 g), providing more than 15% RDA. Vitamin B12(1.27 μg/100 g), niacin (5.62 mg/100 g), and pyridoxine (0.44 mg/100 g) were able to meet 127, 31, and 22% RDA, respectively. Total collagen was 1.22 g/100 g wet weight, 68% of which was soluble. Xanthine was never detected. Adenine, guanine, and hypoxanthine mean levels were 15.47, 9.94, and 92.71 mg/100 g wet weight, respectively.

In this study we investigated the presence of lymphocyte function-associated antigen-1 (LFA-1) and intercellular adhesion molecule-1 (ICAM-1) on renal cell cancer in 16 patients who underwent nephrectomy. In all cases, normal renal tissues presented ICAM-1 on the parietal epithelial cells lining Bowman's capsule while neither the visceral epithelium nor the glomerular endothelium were stained. The epithelial proximal and distal tubular cells were stained poorly (5 +/- 3 positive tubular cross sections/mm2). No renal cells were stained by the monoclonal antibody for LFA-1. In contrast, in renal cancer tissue we found 6 cases, negative for ICAM-1 and the others positive with a mean number of positive tubular cross sections of 52 +/- 37/mm2. In 10 cases staining with LFA-1 antibody was negative, while for the other cases the mean number of positive tubular cross sections was 11 +/- 8/mm2. Our results suggest that the expression of ICAM-1 and LFA-1 on the surface of malignant cells may explain the unusual clinical aspects of renal cell carcinoma, its association with the immune system, and clarify certain aspects of immunotherapy.

Proliferative and necrotizing pneumonia (PNP) is a form of interstitial pneumonia that occurs in weaning and post-weaning pigs. PNP is characterized by hypertrophy and hyperplasia of type II pneumocytes and coagulative necrosis and granular debris within alveolar spaces. Canadian and European studies suggest that the porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are the main causes of the disease, but Aujezsky’s disease virus (ADV) and swine influenza virus (SIV) have also been considered as potential aetiological agents. An immunohistochemical study was carried out on the lungs of 28 Italian pigs with PNP in order to evaluate the role of PRRSV, PCV2 and ADV in PNP lesions. PRRSV infection was identified in the lungs of 11 pigs, PCV2 in the lungs of four pigs and coinfection with both viruses in the lungs of eight pigs. Neither virus was detected in the lungs of the remaining five pigs. ADV antigen was not detected in any sample. The principle aetiological agent of PNP in Italy therefore appears to be PRRSV. Coinfection with PRRSV and PCV2 is characterized by more severe microscopical changes in affected lungs.

The aim of the study was to investigate the effects of supplementation of a microencapsulated blend of tributyrin and lactitol (TL) to a standard Eu- ropean (EU) diet without antibiotic growth promoters on intestinal metabolism and mucosa development of weaned piglets and to compare it with a standard US diet containing animal proteins, zinc oxide, copper sul- fate, and carbadox. Ninety piglets weaned at 21 d were divided into 3 dietary groups consisting of 5 replicates each: 1) US diet supplemented with 55 mg/kg of carba- dox, and 2.5% each of plasma proteins and spray-dried blood cells in the first phase, 3,055 mg/kg of Zn in the first and second phases, and 180 mg/kg of Cu in the third phase; 2) EU diet based on vegetable proteins and no antibiotics; and 3) the same EU diet supplemented with 3,000 mg/kg of microencapsulated TL. The study was divided into 3 phases: 0 to 7, 8 to 21, and 22 to 35 d. On d 7, 21, and 35, animals were weighed, and feed consumption and efficiency were determined. On d 14 and 35, one pig per pen was killed, and the intes- tinal contents and mucosa from the proximal, middle, distal jejunum and the ileum were sampled. Intestinal wall sections were fixed for histological analysis, and intestinal content was used for VFA, ammonia, and polyamine analysis. Throughout the study (d 0 to 35), the US diet had greater ADG and ADFI than the EU diet (P < 0.05). The EU diet supplemented with TL tended to have 11% greater ADG (P = 0.17). Feeding the EU diet caused a reduction in proximal and middle jejunum villi length by 10% (P < 0.05) and an increase in crypt size in proximal jejunum (P < 0.05) compared with the US diet, probably due to an increased rate of cell loss and crypt cell production. The TL supplemen- tation resulted in longer villi along the jejunum and less deep crypts in the proximal jejunum (+15.9 and -8.9%, respectively; P < 0.05) than the unsupplement- ed EU diet. The TL diet increased the concentrations of cadaverine and putrescine in the small intestine (P < 0.05) and seemed to increase cadaverine, histamine, putrescine, and spermine in the large intestine by 1.5- to 10-fold compared with the US or EU diet. In con- clusion, although the US diet had a greater effect on growth performance and mucosal trophic status than the EU diets, the supplementation with slowly released TL seemed to be an effective tool to partially overcome the adverse effects of vegetable protein diets.

Data on the long-term treatment of myeloma bone disease with bisphosphonates are scanty. In a prospective pilot trial we evaluated the effect of long-term parenteral administration of dichloromethylene bisphosphonate (Clodronate), in addition to standard chemotherapy, in 30 patients with active myeloma bone disease. Patients were treated with a mean of 4 courses (range 2-8) of Clodronate: 300 mg/day i.v. for seven days followed by 100 mg/day i.m. for 10 days, administered at a mean interval of 4 months (range 3-6). The median follow-up was 24 months (range 8-36). Clodronate reduced bone pain rapidly and significantly, and reduced the mean values of the biochemical indices of bone resorption to within normal limits; these effects were maintained throughout the follow-up. In three hypercalcemic episodes serum calcium became normal after 2-5 days of treatment with Clodronate. No toxic or side effects were noticed. The occurrence of skeletal morbidity in patients treated with Clodronate was compared with that observed in the control group of myeloma patients (p less than 0.001) in severe bone pain as well as in the incidence of new osteolytic lesions and pathological fractures (p less than 0.001). Supportive Clodronate therapy contributes significantly in controlling the progression of myeloma bone disease.

Sclerotherapy for hydroceles was performed in 18 patients. Cutaneous anaesthesia was induced with an anaesthetic cream (lidocaine and prilocaine, EMLA cream) and a fibrin sealant (Tissucol) was injected into the sac after fluid aspiration. Patients experienced no pain during needle insertion and sclerosant procedure; 2 recurrences were observed during follow-up. EMLA cream anaesthesia and fibrin adhesive sclerotherapy represent a useful alternative to surgical treatment of hydroceles.

Gleason score has been identified as an important variable to predict disease extent and biologic behaviour of prostate cancer. However, the correlation between Gleason score of needle biopsy and surgical specimen is often poor. We studied 72 patients who underwent needle biopsy and radical prostatectomy to correlate Gleason score with PSA, clinical and pathological tumour stage. Only 47.2% of Gleason scores were identical in the biopsy and specimens, 37.5% were undergraded and 15.2% were overgraded. Correlations between clinical and pathological stage were identical in 30.5% of patients, 61.1% of patients were understaged and 8.3% overstaged. In conclusion, accuracy of clinical staging and grading of prostate cancer is low. Although the Gleason score on needle biopsy might be useful to predict the final stage and grade, correlation with surgical specimen is poor.

To compare and assess the function of ileal neobladders with different reconfiguration and with several types of ureteral reimplantation.Forty-five male patients underwent radical cystectomy and detubularized ileal neobladder. In 20 patients an ileal neobladder was carried out according to Studer's technique, in 12 a 'W'- and in 13 a 'U'-shaped neobladder. In the Studer's patients 60 cm of ileum was used, in the 'W' 40 and in the 'U' 30 cm. For the uretero-ileal anastomosis Nesbit's technique was utilized in the Studer's, in the 'W' and 'U' neobladders Camey Le Duc's technique was performed instead. Four patients underwent a serous-lined extramural tunnel reimplantation. Follow-up included a functional and morphological study of the urinary system and a urodynamic study.All Nesbit's uretero-ileum anastomoses resulted refluent when the reservoir was filled up, 15 of 50 ureteral reimplantations according to the Camey Le Duc technique showed reflux at full filling. At 3, 6 and 12 months follow-up, the double reconfiguration reservoirs (Studer's and 'W') showed a larger capacity and a lower maximum pressure than neobladders with a single bending. At 12 months, continence and the voiding interval time was significantly higher in the double reconfiguration than in the 'U' neobladders.The double reconfiguration of the reservoir ('W') might be preferable to that with a single one. As for the type of ureteral anastomosis to select, the problem is still debatable even if in our case-control study we have had better results in terms of reflux and stenosis with the uretero-enteric anastomosis with Nesbit's and associated afferent long tubular ileal limb than with Studer's technique.

A total of 208 patients with superficial transitional-cell carcinoma of the bladder (STCC) after transurethral resection were treated with 30 mg intravesical instillations of mitomycin C (MMC) weekly for 8 weeks, followed by monthly maintenance doses for 12 months. All patients were controlled with a urinary cytological examination every 2 months and with cystoscopy every 3 months. Mean follow-up was 47.8 and 49.3 months in the prophylaxis and control groups, respectively. The incidence of tumor recurrences at the 12th and 48th months was 29 and 44%, respectively, in the MMC group and 45 and 58%, respectively, in the control group. Progression evaluated by grade and stage was significantly higher in the control group. These data indicate that MMC appears to be effective in the prophylaxis of STCC, but the possibility of long-term relapse suggests maintenance of a longer therapy.

Thirty-nine patients were evaluated after retropubic prostatectomy, by urodynamic studies. They were divided into three groups depending upon the degree of urinary continence. A statistically significant difference was found between different groups in mean functional profile length and maximal urethral closure pressure. Differences in age, previous prostatic surgery, tumour extension, or preservation of the neurobundles did not have any significant influence on recovery of continence.

Twenty patients with sexual inadequacies secondary to different degrees of chronic renal failure were investigated. The patients were divided into two groups: group A—ten patients treated with a conservative diet therapy; group B—ten patients on hemodialysis. All the patients under investigation were administered 2000 IU of hCG every other day for 120 days after a 30-day period without treatment. Andrologic, biometric, hematologic, and sexual data were evaluated before and after hormone treatment. Sexual parameters were collected through self-evaluation by the patients and their partners. Our results agree with Van Kammen et al (1975) and show an improvement of the andrologic and biometric data in patients from group A. Furthermore, eight patients form group A and three patients from group B reported a remarkable improvement in their sexual responses.

Nine cases of renal hydatid disease are reported. All diagnostic procedures are discussed and the important role of ultrasound renal scan and cyst puncture is emphasized. Although it is rare, renal secondary location has to be suspected in cases of hepatic echinococcosis and conservative surgical treatment is required to preserve renal function.

The results obtained in a series of 233 subjects who underwent surgery for total cystectomy and ureterosygmoidostomy for carcinoma of the bladder are analyzed. The immediate and long-term results, the complications and length of survival, both in general and in relation to the stage of the neoplasia, have been considered. The surgical death rate was 7%. Survival, taking all cases into account, was 80% after 1 year, 55% after 3 years, 36% after 5 years and 8.3% after 10 years. Significant differences in survival were found among those patients with pathologic stage PIS-P1-P2 and those with P3-P4. From a survival of 64% after 5 years in the first group, we pass to 23% for P3 and 0% for P4. The precocious complications are anastomosis stenosis (1.8%), uro-fecal fistulae (1.8%) and urinary fistulae (1.4%). Late complications are anastomosis stenosis (13.6%), pyelonephritis with episodes of fever (19.3%), severe acidosis (1.8%) and kidney stones (1.8%).

• We have compared in an open trial the clinical and biochemical effects of a new aminodiphosphonate, aminohydroxybutylidene diphosphonate, with those of dichloromethylene diphosphonate, which has been proved effective. The patients presented extensive and symptomatic bone involvement from multiple myeloma, breast cancer, and other metastatic tumors. The treatment consisted of aminohydroxybutylidene diphosphonate, 2.5 mg/d intravenously for five days, or dichloromethylene diphosphonate, 300 mg/d intravenously for seven days, followed by 100 mg/d intramuscularly for ten days. Twelve patients treated with aminohydroxybutylidene diphosphonate and 16 patients treated with dichloromethylene diphosphonate were assessable and were followed up for one to six months. Therapy with aminohydroxybutylidene diphosphonate showed a quicker action in reducing bone pain and reduced significantly more the serum calcium level than did therapy with dichloromethylene diphosphonate. Aminohydroxybutylidene diphosphonate therapy also affected urinary calcium levels and hydroxyproline excretion more markedly than did dichloromethylene diphosphonate, although the differences are not statistically significant. However, the biochemical indexes rebounded more quickly in patients treated with aminohydroxybutylidene diphosphonate, indicating that the loading amount (only 12.5 mg) used in this preliminary study is insufficient to sustain a prolonged effect. The effectiveness and lack of side effects render aminohydroxybutylidene diphosphonate an attractive treatment for malignant bone resorption. ( Arch Intern Med 1987;147:1629-1633)

A case of a forty-five-year-old woman with fused kidneys and intercommunicating pelves drained by a single ureter crossing the midline is reported. A review of the literature is presented.

Samples of superficial inguinal and bronchial lymph nodes, ileum, tonsil and lung were taken from three to five pigs on each of 61 farms with a clinical history of postweaning multisystemic wasting syndrome (PMWS). The samples were examined histologically and by immunohistochemistry for porcine circovirus type 2 (PCV-2). PMWS was diagnosed in two stages: first, an evaluation of the haematoxylin and eosin-stained sections that identified the cases in which the characteristic PCV-2 cytoplasmic inclusion bodies were apparent, and secondly, a conclusive step in which immunohistochemistry was applied to confirm PMWS in the cases in which there were positive immunohistochemical results that coincided with lesions indicative of PMWS in at least one of the lymphoid and/or lung tissues. The location of PCV-2 in specific lesions (cell depletion in lymphoid organs and interstitial pneumonia) confirmed PMWS in 45 of the 61 farms, 31 of which were also infected with porcine reproductive and respiratory syndrome virus. The lymphoid tissues were more reliable than the lungs for the diagnosis of PMWS, both in individual pigs and in groups of pigs, and farm diagnoses based on a group of pigs were more reliable than diagnoses based on single pigs.