Improving the accuracy of pancreatic cancer clinical staging by exploitation of nanoparticle-blood interactions: A pilot study.

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) early diagnosis is  crucial  and new, cheap and user-friendly techniques for biomarker identification  are  needed. "Protein corona" (PC) is emerging a new bio-interface potentially useful in tumor early diagnosis. In a previous investigation, we showed that relevant differences between the  protein patterns of  PCs formed on lipid NPs after exposure to PDAC and non-cancer plasma  samples exist. To extend that research, We performed this pilot study to investigate the effect of PDAC tumor size and distant metastases on PC composition. METHODS: Twenty PDACs were clinically staged according to the UICC TNM staging system 8 t h Edition. Collected plasma samples were let to interact with lipid NPs; resulting PCs were characterized by SDS-PAGE. To properly evaluate changes in the PC, the protein intensity profiles were reduced to four regions of molecular weight: < 25 kDa, 25-50 kDa, 50-120 kDa, > 120 kDa.  RESULTS: Data analysis allowed to distinguish T1-T2 cases from T3 and above all from metastatic ones (p < 0.05). Discrimination power was particularly due to a subset of plasma proteins with molecular  weight comprised between 25-50 kDa  and 50-120 kDa. CONCLUSIONS: PC composition is critically influenced by tumor size and presence of distant metastases in PDAC. If our findings will be further confirmed, we envision that future developments of cheap and user-friendly PC-based tools will allow to improve the accuracy of PDAC clinical staging, identifying among resectable  PDACs with potentially better prognosis (i.e. T1 and T2) those at higher risk of occult distant metastases.

Data of postoperative complications related to fibrinogen-to-albumin ratio in pancreatic resections.

Pancreatic surgery is one of the surgeries burdened with the highest mortality and morbidity rate. This is due both to the aggressive biological nature of the pathology affecting the organ and to the technical difficulties associated with surgery. A further aspect on which research is focusing is represented by inflammation related to oncological pathology. Inflammation plays an important role in tumor progression, and growing evidence has confirmed that the fibrinogen-to-albumin ratio (FAR) is an important prognostic factor for overall survival (OS) in malignant tumors. Inflammatory markers had demonstrated also a role in the prediction of postoperative complication after pancreatic surgery. We speculate that FAR, as an easily available, cost-effective, and non-invasive prognostic indicator for pancreatic cancer patients, could help to identify patients at increased risk of postoperative pancreatic fistula (POPF). We therefore retrospectively analyzed the data relating to 117 pancreatic resections relating direct and indirect markers of inflammation with the incidence of post-operative complications.

PTCH1 gene haplotype association with basal cell carcinoma after transplantation.

BACKGROUND: Basal cell carcinoma (BCC) is 10 times more frequent in organ transplant recipients (OTRs) than in the general population. Factors in OTRs conferring increased susceptibility to BCC include ultraviolet radiation exposure, immunosuppression, viral infections such as human papillomavirus, phototype and genetic predisposition. The PTCH1 gene is a negative regulator of the hedgehog pathway, that provides mitogenic signals to basal cells in skin. PTCH1 gene mutations cause naevoid BCC syndrome, and contribute to the development of sporadic BCC and other types of cancers. Associations have been reported between PTCH1 polymorphisms and BCC susceptibility in nontransplanted individuals. OBJECTIVES: To search for novel common polymorphisms in the proximal 5' regulatory region upstream of PTCH1 gene exon 1B, and to investigate the possible association of PTCH1 polymorphisms and haplotypes with BCC risk after organ transplantation. METHODS: Three PTCH1 single nucleotide polymorphisms (rs2297086, rs2066836 and rs357564) were analysed by restriction fragment length polymorphism analysis in 161 northern Italian OTRs (56 BCC cases and 105 controls). Two regions of the PTCH1 gene promoter were screened by heteroduplex analysis in 30 cases and 30 controls. RESULTS: Single locus analysis showed no significant association. Haplotype T(1686)-T(3944) appeared to confer a significantly higher risk for BCC development (odds ratio 2.98, 95% confidence interval 2.55-3.48; P = 0.001). Two novel rare polymorphisms were identified at positions 176 and 179 of the 5'UTR. Two novel alleles of the -4 (CGG)(n) microsatellite were identified. No association of this microsatellite with BCC was observed. CONCLUSIONS: Haplotypes containing T(1686)-T(3944) alleles were shown to be associated with an increased BCC risk in our study population. These data appear to be of great interest for further investigations in a larger group of transplant individuals. Our results do not support the hypothesis that common polymorphisms in the proximal 5' regulatory region of the PTCH1 gene could represent an important risk factor for BCC after organ transplantation.

Activity of telithromycin against multi-drug resistant Streptococcus pneumoniae and molecular characterization of macrolide and tetracycline resistance determinants.

The antistreptococcal activity of telithromycin and 11 different comparators was evaluated in 26 multi-drug resistant (MDR) Streptococcus pneumoniae strains collected during 2002-2003 as part of the ongoing PROTEKT (Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin) Italian Surveillance Program. The strains were characterized for their susceptibility to antibiotics both at the phenotypic and genotypic levels; furthermore, the association of erm(B), mef(A) class and tet(M) genes, as well as the mobile elements carrying them were determined. The strains in this study were resistant to penicillin (MIC > or = 2 mg/l) in 23.1% of cases, resistant to tetracycline in 88.4%, to cotrimoxazole in 34.6% and cefuroxime in 26.9% while only telithromycin and levofloxacin retained 100% activity against all microorganisms. Co-existence of different resistance determinants was found in 19.2% of all isolates collected in our laboratory, coming from southern Italy. Twenty-three isolates showing the MLSB phenotype of resistance possessing the erm(B) gene (88.5%), associated with tet(M), were carried on the same Tn1545-like element, while two isolates showing the M phenotype possessing the mef(A) gene alone, were carried on Tn1207.1. In only one strain were mef(E) and tet(M) together carried on Tn2009.

Is heartworm disease really spreading in Europe?

Based on recently published surveys and newly acquired data, a study was conducted to verify the distribution of filarial worm (Filarioidea) infections in Europe, with particular emphasis on canine heartworm infection (Dirofilaria immitis). A Geographic Information System based on thermal regimen was constructed as a means to identify areas potentially suitable for heartworm transmission, taking into account that the development of D. immitis larvae in the mosquito does not occur below the threshold temperature of approximately 14 degrees C. Furthermore, a bionomic model of D. immitis in its mosquito vectors, which calculates the moving cumulative heartworm development unit parameter, was applied using the available temperature data to assess the theoretic transmission timing of heartworm in Europe. The results show that the earliest infection risk occurs in Spain on March 21 and the latest risk occurs in Spain on September 11. The longest risk period occurs in Spain (Murcia station: March 21-November 11), and the shortest risk period occurs in northeastern Europe. The study also provides the first risk assessment maps for Europe and suggests that if the actual climatic trend continues, filarial infection should spread into previously infection-free areas.

A classification model of grazing areas in southern Italy using Remote Sensing data.

Detection of vegetation typologies is of particular interest in epidemiological studies on animal diseases. This paper is focused on the automatic detection of grazing areas in southern Italy browsed by equines, oxen and sheep, mostly sedentary, with many in fenced areas for overnight stays. Results revealed Satellite Remote Sensing was an indispensable tool in area surveys for vegetation cover characterization. Our classification model shows an accuracy level of 90.21% and a precision of 92.69%. Results suggest similar application protocols can be used in areas with different vegetation cover to characterizing potential infection risk areas for geohelminths and other diseases.

Reverse transcription polymerase chain reaction method for the detection of glycopeptide resistance in enterococci.

In this work we have developed reverse transcription polymerase chain reaction (RT-PCR) methods for detecting specific mRNA from enterococci, particularly vanA and vanB genes, responsible for glycopeptide resistance in this genus. mRNA from the two genes was detected immediately after RNA extraction of a midlog phase culture, determined by growth rate analysis. Because of the short half-life associated with many bacterial RNA species (1.5-2 min), time is an important factor in obtaining RNA of good yield and high purity. Our results showed that: (i) the transcription of mRNA related to vanA ligase in enterococci showing Van A phenotype happens only after induction with both vancomycin and teicoplanin; (ii) the transcription of mRNA related to vanB ligase happens only in the presence of vancomycin and (iii) there was no transcription of mRNA in the two strains positive to vanA gene after PCR experiments. RT-PCR methodology can have numerous applications in microbiology for studying gene expression in isolated bacteria and also in nonculturable cells in environmental samples, for studies of mechanisms and/or as an indicator of viability in bacterial communities.

Vitamin E-coated dialyzers reduce oxidative stress related proteins and markers in hemodialysis--a molecular biological approach.

BACKGROUND: Hemodialysis patients (HD) are exposed to oxidative stress which contributes to cardiovascular disease and accelerated atherosclerosis, major causes of mortality in these patients. A new dialysis membrane coated with vitamin E has been proposed against oxidative stress and atherosclerosis due to their ability to inhibit lipid peroxidation by interacting with scavengers. The mechanisms however are not completely clarified. This study evaluated, using a molecular biology approach, the effect of 6 months treatment with vitamin E-modified dialyzers, CL-E, on the gene expression of oxidative stress related proteins and markers. PATIENTS AND METHODS: To this end, the gene expression of p22phox, a NAD(P)H oxidase subunit closely linked with the generation of superoxide anions and of Heme oxygenase-1 (HO-1), induced by and protective from oxidative stress, were evaluated by RT-PCR in mononuclear cells from 5 patients under 3 times a week chronic bicarbonate dialysis. Hydroperoxide (HPO) and total antioxidant power (AOP) plasma levels were evaluated at 3 and 6 months of treatment. HPO was also evaluated in 8 patients under CL-E treatment for 1 year and compared with 8 patients treated with cuprammonium-ryon filter (TAF). RESULTS: p22phox mRNA decreased from 0.61 +/- 0.05 d.u. to 0.48 +/- 0.03, p < 0.01 while HO-1 increased from 0.55 +/- 0.04 d.u. to 0.62 +/- 0.03, p < 0.01. HPO decreased in CL-E treated patients: from 2.72 +/- 0.26 microM to 1.45 +/- 0.27 at 3 months (p < 0.001) to 0.87 +/- 0.11, p < 0.001 at 6 months, while AOP increased: from 752 +/- 90 mmol/L to 1057 +/- 105, p < 0.001 at 6 months. HPO was also reduced in 1 year Excebrane CL-E treated patients compared with cuprammonium treated patients: 2.25 +/- 0.3 vs. 1.42 +/- 0.11 microM, p < 0.001. CONCLUSION: The reduced expression of oxidative stress related proteins and markers gives further support to the efficacy of the use of Vitamin E coated dialysers for the prevention or slowing progression of cardiovascular disease and atherosclerosis, major complications and causes of mortality in these patients in which oxidative stress plays a pivotal role.

Use of remote sensing and geographical information systems to identify environmental features that influence the distribution of paramphistomosis in sheep from the southern Italian Apennines.

A geographic information system (GIS) was constructed using remote sensing (RS) and landscape feature data together with Calicophoron daubneyi positive survey records from 197 georeferenced ovine farms with animals pasturing in a 3971 km(2) area of the southern Italian Apennines. The objective was to study the spatial distribution of this rumen fluke, identify environmental features that influence its distribution, and develop a preliminary risk assessment model. The GIS for the study area was constructed utilizing the following environmental variables: normalized difference vegetation index (NDVI), land cover, elevation, slope, aspect, and total length of rivers. These variables were then calculated for "buffer zones" consisting of the areas included in a circle of 3 km diameter centered on 197 farms. The environmental data obtained from GIS and RS and from data taken by the veterinarians on the field (stocking rate and presence of streams, springs and brooks on pasture) were analyzed by univariate (Spearman and ANOVA) and multivariate (discriminant) statistical analyses using the farm coprological status (positive/negative) as the dependent variable. Sheep on 32 of the 197 (16.2%) farms, were positive for C. daubneyi, with an average intensity of 52 epg. A multivariate stepwise discriminant analysis model was developed that included moors and heathland, sclerophyllous and coniferous forest vegetation, autumn-winter NDVI and presence of streams, springs and brooks on pasture. The variables entered in the model were also correlated with C. daubneyi positive farms in the univariate tests and are consistent with the environmental requirements of C. daubneyi and its snail intermediate host.

Treatment of secondary hyperparathyroidism with low dose intermittent calcitriol in hemodialysis patients. Imaging and cost analysis.

Calcitriol therapy is effective in the treatment of secondary hyperparathyroidism both during intravenous and oral administration, but there are doubts about the length of therapy and the duration of results. There are conflicting reports about results in size and activity of enlarged glands studied by ultrasound and double-tracer-subtraction-scintigraphy (DTSS). In 12 patients, 1 microgram of calcitriol was administered three times a week, intravenously and orally in alternate modes, for 46 weeks (therapy period) and orally for 46 weeks (follow-up period). During therapy, parathyroid hormone levels decreased in all patients, and in eight decreased by about 50% and were maintained at low levels during follow-up in five patients. Nine enlarged glands were detected by ultrasonography at the start of the study, and four hotspots were detected by DTSS; ultrasonography and DTSS were repeated at the end of the therapy and at the end of the follow-up: ultrasonography did not yield any significant variation in size, while one hot spot disappeared on DTSS. Basing their judgment on the lower cost of oral rather than intravenous administration, and on the good results of oral therapy, the authors stress the advisability of taking into account clinical and financial considerations before choosing the route of administration.

[Geographical Information Systems and remote sensing technologies in parasitological epidemiology]. / I GIS (Geographical Information Systems) e la tecnologia satellitare nella epidemiologia delle parassitosi.

Parasites have natural habitats in the same way as a species: they are found in focal areas where the spatial distribution of the parasite, host, vector and required environmental conditions coincide. The spatial distribution of parasites is, therefore, a function of the interaction between abiotic and biotic environmental factors. The boundaries of distributions are not strictly fixed in space and time and may fluctuate with climate and other components of the environment or anthropical factors. Geographic Information Systems (GIS) and remote sensing (RS) technologies are being used increasingly to study the spatial and temporal patterns of disease. GIS can be used to complement conventional ecological monitoring and modelling techniques, and provide means to portray complex relationships in the ecology of disease. In addition, the use of GIS and RS to identify environmental features allows determination of risk factors and delimitation of areas at risk, permitting more rational allocation of resources for cost-effective control. Since 1996, GIS have been used in our territorial cross-sectional and longitudinal parasitological surveys in order to experiment new applications to plan sampling protocols and to display quickly, clearly, and analytically the spatial and/or temporal distribution of parasitological data. The use of GIS allowed us to draw the following types of descriptive parasitological maps: distribution maps, distribution maps with proportioned peaks, choroplethic maps with proportioned peaks, point distribution maps and point distribution maps with proportioned peaks. In a recent study, GIS and RS technologies have been used also to identify environmental features that influence the distribution of paramphistomosis in sheep from the southern Italian Apennines and to develop a preliminary risk assessment model. A GIS was constructed using RS and landscape feature data together with paramphistome positive survey records from 197 georeferenced ovine farms with animals pasturing in an area of the southern Italian Apennines. The GIS for the study area was constructed utilizing the following environmental variables: Normalized Difference Vegetation Index (NDVI), land cover, elevation, slope, aspect, and total length of rivers. In addition, data regarding the presence of watercourses smaller than rivers, namely, streams, springs and brooks were recorded in the field. All these variables were then calculated for "buffer zones" consisting of the areas included in a circle of 3 Km diameter centred on 197 farms. The environmental data obtained were analyzed by univariate and multivariate statistical analyses using the paramphistome farm coprological status (positive/negative) as the dependent variable. A multivariate stepwise discriminant analysis model was developed that included moors and heathland, sclerophyllus and coniferous forest vegetation, autumn-winter NDVI and presence of streams, springs and brooks on pasture. The variables entered in the model are consistent with the environmental requirements of paramphistomes and their snail intermediate host. In particular, the land cover types entered in the model in this area are indicators of marginal uncultivable and sloping zones where typically there is the presence of water (permanently or temporarily). In addition, since NDVI can be used as an indicator of regional thermal-moisture regime, the distribution of farms positive for paramphistomosis corresponding to relatively high values of winter NDVI indicated the presence of adequate moisture and temperatures favourable to the rumen fluke and the snails. In conclusion, GIS and RS are useful to define the habitats of parasites, especially for those with strong environmental determinants, and to produce forecasting maps requested for the planning and the monitoring of control strategies on small and large scale.

[Radioimmunologic determination of circulating parathyroid hormone in patients with primary and secondary hyperparathyroidism]. / Determinazione radioimmunologica del paratormone circolante in pazienti con iperparatiroidismo primitivo e secondario.

A radioimmunological evaluation was made of plasma parathormone (PTH), using an antibody against the C-terminal fraction, in 14 subjects with primary parahyperthyroidism, and in 19 anuric subjects with secondary forms on three-weekly dialysis. In all cases, values were significantly higher (p less than 0.01) than in a group of normal controls. In secondary forms, the same significance was also apparent on comparison with primary forms, probably due to an absence of renal clearance of the hormone.

[Physical training and low-dose heparin-calcium in patients suffering from chronic obliterating arteriopathy of the lower limbs with intermittent claudication]. / Training fisico ed eparina calcica a basse dosi in pazienti affetti da arteriopatia obliterante cronica degli arti inferiori con claudicatio intermittens.

In the treatment of peripheral obliterating arteriopathy (POA) physical training provides clear results in terms of increased walking autonomy; it is still not fully clear whether the positive effects of physical training can be further improved by concomitant back-up drug therapy. For this purpose 374 patients of both sexes, with a mean age of 64 years, suffering from chronic peripheral obliterating arteriopathy of the lower limbs were enrolled in a controlled open clinical trial, instructed to follow a programme of physical training and randomly allocated to low dose treatment with heparin calcium (12,500 IU/day) for 6 months. An improvement in the claudicometric parameters (free gait interval, absolute gait interval and recovery time) measured at constant speed and in the resting Winsor ankle/arm index of the most severely damaged limb were observed in both groups. These improvements were significantly greater in the group receiving pharmacological treatment (p < 0.01) and efficacy increased in line with basal deambulatory impairment. The results obtained and the good tolerance of the drug underline the clinical efficacy of heparin calcium at low doses in association with a physical training programme in patients suffering from Fontaine's stage II peripheral obliterating arteriopathy.

[Spontaneous rupture of the esophagus]. / La rottura spontanea dell'esofago.

Spontaneous rupture of the esophagus is a rare emergency that requires immediate diagnosis and superior surgical treatment. Delay in diagnosis is common and is caused mostly by lack of consciousness of the disease and consequent tendency to diagnose a cases as perforated peptic ulcer, pancreatitis, acute coronary disease etc. Mortality and morbidity increase proportionally to the time between esophageal perforation and adequate treatment. In this paper the Authors present a case of Boerhaave's syndrome in a patient who underwent two surgical repairs followed by leakages of the esophageal sutures. The definitive successful treatment has been obtained with a third operation performed after complete sterilization of the mediastinum.

[Correlation between methicillin-resistance and resistance to fluoroquinolones in Staphylococcus aureus and Staphylococcus epidermidis]. / Correlazione della resistenza alla meticillina e ai fluorochinoloni in Staphylococcus aureus e Staphylococcus epidermidis.

Fluoroquinolones resistance in Staphylococci is associated to point mutations in grlA (80,84 and 116) grlB, gyrA (84,88) and gyrB genes. Almost all MRSA strains are ciprofloxacin and levofloxacin resistant while, in a lesser degree, MRCoN staphylococci show to be resistant to levofloxacin. This observation made possible to predict a different correlation between methicillin-resistance and the resistance to FQs in this two different species. In this study, we compare genomic analysis of S. aureus and S. epidermidis with the resistance to FQs. Our results show that strains of MRSA are distributed in 4 different PFGE-types while 12 MRSE strains are distributed in 9. MRSA resistant to FQs showed a unique PFGE pattern; on the contrary of FQs susceptible MRSA and MSSA. Furthermore mecA and gyrA genes are located in the same SmaI fragment in MRSA and in different in MRSE. MSSE and MRSE show more ClaI/mecA polymorphisms than MRSA. All this data confirm the clonal origin of MRSA and show that FQs resistance is linked to the presence of mec locus and both clonally spread. On the contrary in MRSE FQs-resistance is independent from MR and arise with the normal frequence of antibiotic induction.

[Clonal diffusion and evolution of mecA and Tn554 polymorphisms in methicillin-resistant Staphylococcus aureus in Italy]. / Diffusione clonale ed evoluzione dei polimorfirsmi di mecA e Tn554 in Staphylococcus aureus meticillino-resistente in Italia.

The purposes of the present study were to track the geographic spread of 69 MRSA strains in Italy recovered from 7 hospitals in four towns; to detect the clonal identities among the isolates by a combination of multiple genomic typing methods and to measure temporal trends in clonal types between 1984 and 1998. Our results showed the spread of three major clones among the MRSA isolates of 1984-1995 period: the most represented MRSA clone carried the PFGE pattern A, the mecA polymorph II and had no homology with Tn554 (II::NH::A); most of these isolates were susceptible to the macrolides,being similar to the historically " archaic" MRSA strains; the clone typed I::E::A, carried the PFGE pattern A, the mecA polymorph I and Tn554E commonly defined as "Iberian clone"; unique clone, showing an uncommon PFGE pattern E. the mecA polymorph II and the Tn554 E (II::E::E) and were characterized by a uniform susceptibility to tetracycline and rifampin. During 1997-98 the representation of this clone increased instead of the classical "Iberian clone". A new multi-resistant MRSA strain, carrying the PFGE pattern B (or B'), the mecA polymorph XI and Tn554 polymorph B (XI::B::B), called "Brazilian clone", increased from being absent (1984-95) to 48%. Our molecular data show an Italian MRSA "scenario" far from the common European trends and clearly documented the spread of an archaic clonal type (II::NH::A) in 1984-95, the arrival and rapid increase of Brazilian done in 1997-98 and the chronological and geographical spread of a unique (H::E::E) called "Italian clone", instead of the widely spread Iberian MRSA clone.

[Cavoauricular thrombectomy under extracorporeal circulation and right superior lobectomy for paraneoplastic thrombosis in a case of lung cancer]. / Thrombectomie cavo-auriculaire en C.E.C. et lobectomie supérieure droite pour thrombose paraneoplasique dans un cas de cancer du poumon.

A patient with superior vena cava thrombosis from cancer of the right lung is described. The particular interest of this case resides in: The numerical scantiness of cases reported in the literature: 12 cases reviewed; The exceptional nature of the etiopathogenic mechanism: paraneoplastic thrombosis; and the more precise problems relating to the indication, technique and tactics with respect to surgery.

[Cystic dilatation of the biliary tract]. / La dilatazione cistica della via biliare.

Cystic dilatation of the biliary tract is a rare disorder, more common in children, with a high morbidity and mortality rate. Death may follow one of the possible complications (recurrent cholangitis, biliary cirrhosis, cholangiocarcinoma). The Authors report the case of an asymptomatic adult woman. According to CT scan and ERCP the lesion found was classified as type IVb of Todani's classification. The cyst was removed and a Roux en-Y reconstruction was performed. This procedure is currently associated to a low mortality rate and the best long term results.

[Rare tumor of the stomach: leiomyoblastoma]. / In tema di tumori rari dello stomaco: il leiomioblastoma.

The Authors report a case of gastric leiomyoblastoma presenting a dyspeptic and painful symptomatology and microcytic hyposideremic anaemia. The patient underwent elsewhere gastric resection and was discharged with the diagnosis of "gastric leiomyoma characterized by bizarre cells". After a review of the literature, the Authors emphasize the anatomo-pathological and clinical aspects pointing out the diagnostic difficulties in this malignancy. The correct treatment is based on predicted factors of malignancy.

[Collagenous colitis: a disease difficult to define]. / La colite collagena: una patologia di difficile definizione.

The Authors report a case with watery chronic diarrhoea not related to malabsorption or to inflammatory bowel disease or to intestinal neoplasm. Colonic biopsy, performed during colonoscopy, with the histologic finding of collagenous material in the subepithelial space associated to epithelial alterations and to the presence of inflammatory cells, confirmed the diagnosis of this rare pathologic condition. The pathogenesis of the syndrome, however, is still unknown.