Krill oil, vitamin D and Lactobacillus reuteri cooperate to reduce gut inflammation.

Current research into original therapies to treat intestinal inflammation is focusing on no-drug therapies. KLD is a mixture of krill oil (KO), probiotic Lactobacillus reuteri (LR), and vitamin D (VitD3). The aim of this study was to assess in vitro and in vivo the potential cooperative effects of KLD in reducing gut inflammation. Colorectal adenocarcinoma cell lines, CACO2 and HT29, and C57BL/6 mice were used for in vitro and in vivo analyses, respectively. Cells were exposed to cytomix (interferon gamma + tumour necrosis factor alpha (TNF-α)) to induce inflammation or co-exposed to cytomix and KO, LR and VitD3 alone or to cytomix and KLD. Animals were treated for 7 days with dextran sodium sulphate (DSS) to induce colitis or with DSS and KLD. In vitro assays: F-actin expression was analysed by immunofluorescence; scratch test and trans-epithelial electric resistance test were performed to measure wound healing; adhesion/invasion assays of adhesive and invasive Escherichia coli (AIEC) bacteria were made; mRNA expression of TNF-α, interleukin (IL)-8 and vitamin D receptor (VDR) was detected by quantitative PCR. In vivo assays: body weight, clinical score, histological score and large intestine weight and length were estimated; mRNA expression of TNF-α, IL-1ß, IL-6, IL-10 by quantitative PCR; VDR expression was detected by quantitative PCR and immunohistochemistry. In vitro: KLD restores epithelial cell-cell adhesion and mucosal healing during inflammation, while decreases the adhesiveness and invasiveness of AIEC bacteria and TNF-α and IL-8 mRNA expression and increases VDR expression. In vivo: KLD significantly improves body weight, clinical score, histological score and large intestine length of mice with DSS-induced colitis and reduces TNF-α, IL-1ß and IL-6 mRNA levels, while increases IL-10 mRNA and VDR levels. KLD has significant effects on the intestinal mucosa, strongly decreasing inflammation, increasing epithelial restitution and reducing pathogenicity of harmful commensal bacteria.

Fine needle aspiration cytology of thyroid nodules: conventional vs thin layer technique.

OBJECTIVE: Liquid-based cytology using the thin layer technique has recently been introduced in thyroid fine needle aspiration cytology together with or in substitution of direct smears, but its usefulness is still controversial and relatively few studies have been published in this field. The aim of the present study was to compare the results obtained from conventional smears with those from thin layer smears. DESIGN: In 3875 thyroid nodules, a double cytologic sampling was taken in randomized order, to prepare conventional or thin layer smears. MAIN OUTCOME: The diagnoses agreed in 2934 (75.7%) cases and disagreed in 941 (24.3%). The analysis of discordant data showed there were fewer non-diagnostic cases in the thin layer smears (377 vs 541, p<0.001) whereas in conventional smears there were more cases positive for carcinoma (27 vs 4, p<0.001). The cytohistologic correlation was available for 194 cases and showed that conventional smears had a greater capacity for revealing carcinomas (44 vs 31). Finally, diagnoses based on conventional smears were more sensitive than thin layer smears (93.6% vs 65.9%) whereas specificity was constant. CONCLUSIONS: From our experience, the conventional smear offers a greater possibility of diagnosis when suspecting malignancy or diagnosing malignancy cases, whereas thin layer smears significantly reduce the number of non-diagnostic cases. For this reason, we suggest combining the two techniques in routine cytologic diagnosis.

Early admission to stroke unit influences clinical outcome.

An improvement in patient arrival time to stroke unit (SU) is recommended, since earlier stroke management seems to improve 'per se' functional outcome. The objective of this study was to determine if early admission influences the outcome, reduces disability and mortality at discharge and three months later independent of tlirombolytic treatment. Consecutive acute stroke patients admitted to SU between January 1st 2000 and December 31st 2003 were studied in order to analyze the actual role of acute management independent specific pharmacological treatment, we excluded subjects who underwent rt-PA. 35.8% of 2,041 consecutive stroke patients arrived within 3 hours; 62.4% within 6 hours; 37.6% arrived later. Approximately 80% of the <6 hour patients presented a National Institutes of Health Stroke Scale (NIHSS) >4 and modified Rankin Scale (mRS) score >2 in comparison with 60% of the >6 hour patients. In hospital (8.7%) and three-month (7.3%) mortality in <3 hour patients were not significantly different from what observed in >3 hour patients (6.8% and 6.1% respectively) while functional outcome after three months was better in <3 hour patients (NIHSS: 34.6 vs 15.2; mRS: 32.9% vs 16.8%). Old age, history of TIA, cardioembolic etiology, severity of neurological deficit and hemorrhagic stroke type all led to earlier arrival time. Admission within 3 hours 'per se' improves outcome and reduced disability at three months.

Silencing of endogenous IGFBP-5 by micro RNA interference affects proliferation, apoptosis and differentiation of neuroblastoma cells.

Signal transduction through the IGF axis is implicated in proliferation, differentiation and survival during development and adult life. The IGF axis includes the IGF binding proteins (IGFBPs) that bind IGFs with high affinity and modulate their activity. In neuroblastoma (NB), a malignant childhood tumor, we found that IGFBP-5 is frequently expressed. Since NB is an IGF2-sensitive tumor, we investigated the relevance and the function of endogenous IGFBP-5 in LAN-5 and in SY5Y(N) cell lines transfected with micro and small interfering RNAs directed to IGFBP-5 mRNA. Cells in which IGFBP-5 expression was suppressed were growth-inhibited and more prone to apoptosis than the parental cell line and controls. Apoptosis was further enhanced by X-ray irradiation. The ability of these cells to undergo neuronal differentiation was impaired after IGFBP-5 inhibition but the effect was reversed by exposure to recombinant IGFBP-5. Together, these data demonstrate the importance of IGFBP-5 for NB cell functions and suggest that IGFBP-5 might serve as a novel therapeutic target in NB.

Cyclin D1-dependent regulation of B-myb activity in early stages of neuroblastoma differentiation.

Levels of the transcription factor B-myb must be down-regulated to allow terminal differentiation of neuroectodermal cells and yet its constitutive expression induces early markers of neural differentiation. Thus, we investigated potential mechanisms of enhanced B-myb activity in early stages of neural differentiation. We report here that B-myb expression does not decrease, cyclin A and Sp1 levels remain constant while p21 levels increase continuously upon retinoic acid-induced differentiation of the LAN-5 neuroblastoma cell line. In contrast, cyclin D1 expression is down-regulated at the onset of the differentiative process by protein destabilization. Luciferase assays of promoter activity indicate that B-myb-dependent transactivation is enhanced in LAN-5 cells treated with retinoic acid (RA) for 24 h. The enhancement is independent from cyclin A but is suppressed by a degradation-resistant mutant form of cyclin D1. The importance of cyclin D1 in controlling B-myb activity is further suggested by co-immunoprecipitation experiments, showing that the amount of cyclin D1 co-immunoprecipitated with B-myb decreased after RA treatment. Thus, B-myb may play an active role in the early stages of differentiation when its transactivation activity is enhanced as a consequence of cyclin D1 down-modulation.

Computer-assisted immunocytochemical determination of breast cancer steroid receptors. Frozen sections vs paraffin sections.

Previously we have demonstrated that determination of oestrogen (ER) and progesterone (PR) receptors by immunocytochemical assay (ICA) on frozen sections (FS) and cytological smears with image analysis is effective for evaluating steroid receptors. The aim of this study was to determine concordance between ER and PR assessed by ICA on FS and paraffin sections (PS) both evaluated by image analysis. There were 115 breast carcinomas selected. For all cases, ER and PR determination was performed on FS and PS. Computer-assisted image analysis was performed using CAS 200. Results were expressed as percent positive area of neoplastic nuclei compared with total nuclear area of the examined neoplastic cells. Good correlation was demonstrated for both ER (r=0.759; concordance=83.4%) and PR (r=0.800; concordance=87.8%). The unexpected relatively low concordance for ER led to further investigations. We divided the 115 cases in two groups. The first group included specimens from our hospital; the second group specimens from suburban hospitals. In the first group there was better correlation for both ER (r=0.897) and PR (r=0.915) with a concordance of 91.5% and 93.6%, respectively. In the second group, correlation was worse for both ER (r=0.724) and PR (r=0.708), with a concordance of 77.9% and 83.9% respectively. From analysis of discordant cases we conclude that reduction in correlation and concordance with increased false negative cases in group 2 are probably due to delayed fixation. Our data suggest that ICA with automated image analysis is efficient in evaluating ER and PR on paraffin section only when the tumour samples are correctly fixed.

Hemodynamic effects of carbon dioxide insufflation during endoscopic vein harvesting.

BACKGROUND: A prospective study was performed assessing the hemodynamic effects of carbon dioxide (CO2) insufflation during endoscopic vein harvesting (EVH) using the Guidant Vasoview Uniport system. METHODS: Five hemodynamic and respiratory parameters (end-tidal carbon dioxide, arterial partial pressure of carbon dioxide, mean arterial pressure, mean pulmonary arterial pressure, and cardiac output), were measured in 100 consecutive patients undergoing EVH with CO2 insufflation. Data were obtained prior to commencement of EVH, 15 minutes after commencement, and 5 minutes after completion of the vein harvesting. RESULTS: No adverse hemodynamic effects were observed during CO2 insufflation. Specifically, average mean arterial pressure went from 88.77+/-9.64 to 89.13+/-8.60 to 88.24+/-8.71 mm Hg before, during, and after endoscopic vein harvesting (p = 0.291). Likewise, average mean pulmonary artery pressures were 19.76+/-4.75, 20.05+/-4.48, and 20.05+/-4.62 mm Hg (p = 0.547); and average cardiac output was 4.25+/-0.74, 4.22+/-0.73, and 4.23+/-0.69 L/min (p = 0.109) at those three intervals. Additionally, there was no evidence of significant systemic absorption of CO2 as reflected in average arterial PCO2, which remained steady at 37.42+/-5.19, 37.51+/-4.59, and 38.10+/-4.80 mm Hg (p = 0.217); and average end-tidal CO2, which was 32.10+/-3.66, 32.50+/-3.47, and 32.38+/-3.33 mm Hg (p = 0.335). In a subset of 20 patients with elevated pulmonary arterial pressure (more than 32 mm Hg), there was also no significant change in any of the parameters. CONCLUSIONS: Carbon dioxide insufflation during EVH leads to no adverse hemodynamic consequences or systemic CO2 absorption. The technique appears to be safe and well tolerated.

Computer-assisted immunocytochemical determination of breast cancer steroid receptors on cytological smears of excised surgical specimens compared with frozen sections.

BACKGROUND: Due to the widespread use of fine needle aspirate biopsy the practice of determining estrogen (ER) and progesterone (PR) receptors in breast carcinoma from cytological smears (CS) is becoming very common. The aim of this study was to determine concordance between ER and PR assessed by immunocytochemical assay (ICA) on CS and FS both evaluated by image analysis since we have found no data in literature on this. METHODS: 104 breast carcinoma cases were selected. For all cases ER and PR determination was performed on CS, obtained by light scraping of the freshly cut surface of the excised surgical tumors at the time of frozen section diagnosis, and FS using the same monoclonal antibodies. Computer-assisted image analysis was performed in all cases using CAS 200. Results were expressed as percent positive area of neoplastic nuclei compared with total nuclear area of the examined neoplastic cells. RESULTS: Good correlation was demonstrated between percent positive nuclear neoplastic area by ER-ICA on CS and FS (r = 0.759; P < 0.0001). Concordance of results was 90.19% (P < 0.001). Good correlation was also demonstrated between percent positive nuclear neoplastic area by PR-ICA in CS and FS (r = 0.889; P < 0. 0001). Concordance of results was 97.02% (P < 0.0001). CONCLUSIONS: Our data suggest that ICA on CS with automated image analysis is efficient in evaluating ER and PR content in human breast cancer, especially when CS is the only method pathologists have to evaluate receptor status e.g. in advanced breast cancer cases when neoadjuvant therapy is necessary before surgery or when surgery is impossible.

Evaluation of chemically-induced pore surface modifications on rock cores.

Chemical modification of pore surface properties due to active components of crude oils could significantly affect production processes. Specific chemical treatments can prevent surface alteration, protecting pores from deposition of heavy oils components. Both relaxation times analysis and nuclear magnetic resonance (NMR) imaging technique are a valuable method for nondestructive monitoring of solid/liquid interface alteration especially if applied to heterogeneous rock cores, which are impossible to study using conventional methods (e.g., Amott tests and dynamic angle contact). In our work, we examined carbonate core plugs subjected to aging treatments in crude oil. Relaxation times measurement, revealing modifications of fluid/surface interactions and imaging, was applied in characterise heterogeneous systems and reveal fluid distribution changes. Nuclear magnetic resonance results were compared with a special microscopy technique (Cryo-SEM) that allows observation of fluids at the pore level, providing a better understanding of the processes involved.

Estrogen and progesterone receptors in breast cancer: comparison between enzyme immunoassay and computer-assisted image analysis of immunocytochemical assay.

Evaluation of estrogen (ER) and progesterone (PR) receptor content is now an important procedure in the management of breast cancer patients. Production of monoclonal antibodies to ER and PR has permitted development of an enzyme immunoassay (EIA) and immunocytochemical assay (ICA). This study compared the results of ICA and EIA to evaluate ER and PR in 197 breast cancers using the same monoclonal antibodies. The ICA results were obtained by automated computer-assisted image analysis using CAS 200. The cut-off values adopted were 15 fmol/mg protein for EIA and 10% of the positive neoplastic area of the nuclei for ICA. For statistical analysis, Spearman's correlation coefficient and chi 2 were used. There was good correlation between ICA and EIA for both ER (r = 0.714; p < 0.0001) and PR (r = 0.815; P < 0.0001). Of 197 tumors, 136 (69.04%) were ER-ICA+, and 138 (70.05%) were ER-EIA+; 111 (56.35%) were PR-ICA+, and 115 (58.38%) were PR-EIA+. Results were concordant, positive or negative with both methods, in 175 cases for ER and in 173 cases for PR. ER and PR results were only discordant in 22 and 24 cases, respectively. Concordance of results obtained by the two methods was 88.83% (P < 0.0001) for ER and 87.81% (P < 0.0001) for PR. Correlation of results obtained by EIA and ICA to determine ER and PR was good. The data obtained suggest that ICA with automated image analysis is an effective means for evaluating ER and PR content in human breast cancer, especially when, as happens ever more frequently nowadays, the tumor is too small to perform EIA or when retrospective studies are performed.

Neoadjuvant cisplatin and etoposide for stage IIIA (clinical N2) non-small cell lung cancer.

In an attempt to improve the curative potential of surgery, 46 patients with unresectable Stage IIIA (Clinical N2) non-small cell lung cancer received neoadjuvant chemotherapy with cisplatin and etoposide. After 2 or 3 cycles, 45 patients were evaluable for response; the overall response rate was 82% (37/45) with 3 complete and 34 partial responses. Toxicity was primarily hematologic. Surgical exploration was performed on 35 patients, but resection was possible in only 33 (73%). Of these, 28 resections were complete (62%). Four patients (2CR, 2PR; 9%) had no tumor in biopsy specimen. Three deaths were surgery-related. Median survival of the entire 46 patients was 24.5 months with a 2-year survival of 53%. Cisplatin and etoposide is an effective chemotherapeutic regimen for regionally advanced non-small cell lung cancer; the resection and survival rates justify further trials to compare this approach to other treatment modalities.

Diurnal and seasonal variations in the occurrence of stroke: a community-based study.

Several hospital-based studies suggest a circadian rhythm for stroke, with a peak of onset in the morning. To verify whether these results could be confirmed in a community-based study, we used data from the SEPIVAC, a community-based stroke register in Umbria, Italy. Cerebral infarctions (48%), primary intracerebral hemorrhages (54%), subarachnoid hemorrhages (53%) and 'not known' strokes (51%) were all more common between 6 a.m. and noon, and this peak was still present when strokes recognized on awakening were evenly redistributed over the sleep period. Looking at the subgroups of cerebral infarction, lacunar syndromes were more common during sleep. Cerebral infarctions were more frequent during winter, and primary intracerebral hemorrhages during autumn.

SEPIVAC: a community-based study of stroke incidence in Umbria, Italy.

The SEPIVAC study is a community-based epidemiological survey of incidence and outcome of acute cerebrovascular disease in the Sixth Local Health Unit, Umbria, Italy (population 49,218). The study was carried out from 1 September 1986 to 31 August 1989. There were 375 patients who were registered with a first ever stroke, with a crude rate of 2.54 (95% confidence limits 2.29-2.81) per 1000 per year; the rate adjusted to the European population is 1.55 (CL 1.36-1.77). The age adjusted relative risk for males is 1.35 (CL 1.10-1.66). Up to 15% of the patients were not admitted to hospital during the acute phase of their disease. At least 286 (76.3%, CL 72-80.6) of the cases were due to cerebral ischaemia; in 56 of these (19.6%, CL 15-24.2) a clinical diagnosis of lacunar ischaemia was made. The 30 day case fatality rate was 20.3% (CL 16.2-24.3); between one and six months 7.5% (CL 5-10.6) of patients died.

A community-based study of incidence, risk factors and outcome of transient ischaemic attacks in Umbria, Italy: the SEPIVAC study.

The SEPIVAC study is a community-based epidemiological survey of incidence and outcome of transient ischaemic attacks (TIAs) and strokes in the territory of the 6th Local Health Unit, Umbria, Italy, where 49,218 people live, from 1 September 1986 to 31 August 1989. All cases were registered with the study either by notification from general practitioners (GPs) or by a check of hospital admission within the study area and in the two hospitals of Perugia. There were 94 incident cases of TIAs (45 males, 49 females), thus giving a crude rate of 0.64 per 1000 per year [95% conficence intervals (CI) 0.52/0.78]. The rate adjusted to the European population is 0.42 (CI 0.33/0.54). Mean age was 69.4 years, and females were significantly older than males. The weighted relative risk for males was 1.19 (CI 0.79/1.79). Thirty-one patients were treated at home by their GPs. Females had hypertension more frequently than males, whereas males smoked more frequently; we did not find any other statistically significant difference in the distribution of risk factors. Twelve patients out of 58 who had CT had an infarct, and 29 out of 54 submitted to Doppler ultrasonography had carotid stenosis. At 1 month, 4 patients had suffered an ischaemic stroke, 1 of whom died. At 6 months, 3 further strokes and 2 further deaths (1 due to myocardial infarction) had occurred.

Chlorambucil carcinogenesis in BALB/c mice.

Chlorambucil, a drug used in the treatment of neoplastic and non-neoplastic disease, was administered by gavage to BALB/c mice at a dose of 1.0 mg/kg body wt. 5 times per week for 12 weeks to test its carcinogenicity. The survival was statistically reduced in treated animals of both sexes (P less than 0.001). The treatment induced a significant increase in lung tumours (males, P less than 0.001; females, P less than 0.001) and lymphoreticular system tumours (males P less than 0.01; females, P less than 0.001) in both sexes and mammary carcinomas in female mice (P less than 0.05). These results with other investigations reported in literature, suggest that chlorambucil is carcinogenic in laboratory animals, mutagenic and that it could be a potential carcinogenic hazard to man.

5-Fluorouracil carcinogenesis in BALB/c mice.

5-Fluorouracil, a drug mainly used in the treatment of gastrointestinal tract neoplasms, was administered i.p. to BALB/c mice at the dose of 30 mg/kg body weight once a week for 50 weeks to test its carcinogenicity. The treatment induced a significant increase in lung tumor in both sexes (males, p less than 0.05; females, p less than 0.01) and tumors of the lymphoreticular system in female mice (p less than 0.001). These results suggest that 5-fluorouracil is carcinogenic in mice.

[Descriptive epidemiology of malignant tumors of the colon and rectum]. / Epidemiologia descrittiva dei tumori maligni del colon e del retto.

This paper examines patterns and trends of colon and rectal cancer in different countries and in Italy. Incidence and mortality rates of colorectal cancer vary widely in the world. High rates are characteristic of highly developed countries in North America, northern and western Europe. The lowest rates are found in Asia, Africa and most Latin American countries. The most recent incidence rates for colon cancer from cancer registries around the world published in "Cancer Incidence in Five Continents, 1982" range from 0.6 cases per 100,000 in Dakar, Senegal to 32.3 in Connecticut, USA for males and from 0.7, always in Dakar, to 27.4 among the Japanese population of Bay Area, USA. The Italian cancer registry for Varese, shows a rate of 19.9 for males and 16.9 for females. The incidence rates for cancer of the rectum range from 1.5 per 100,000 in Dakar to 22.6 in the North West Territory and Yukon, Canada. For females the highest rates, 13.9, are in Israel (born in Europe or America) and the lowest always in Dakar, 1.0. The Varese rates are 15.7 and 9.1 for males and females respectively. Regression analysis shows that between incidence rates of colon and rectal cancer, divided by sex, there is a strict correlation. The sex ratios for colon and rectal cancer differ, rectal cancer being distinctly more common among males in most countries, whereas colon cancer affecting both sexes at rather similar rates. Results confirm that there is a higher frequency, for colon cancer in particular, in urban areas than in rural areas. Differences due to race, on the contrary, have found no confirmation. The international incidence trends over the period 1960-1980 show a general increase for colon cancer in both sexes. In Asia the most evident increase have been in Singapore and Miyagi. In Europe, Slovenia (Yugoslavia) and Norway present 40-50% increases. Similar tendencies have been observed for rectal cancer trends. Values more than increase 100%, for both sexes, have been found in Hawaii and Singapore. In Europe, Norway and Slovenia always present the highest increases. Marked geographic variations occur even as regards colon and rectal mortality in the different countries. The highest values for colon cancer have been found in Luxemburg (18.4) for males, and in New Zealand (13.3) for females and the lowest in Honduras for both sexes (0.1 and 0.0 respectively). In Italy the values are 8.0 for males and 6.0 for females.(ABSTRACT TRUNCATED AT 400 WORDS)

Tobacco smoking, coffee, cocoa and tea consumption in relation to mortality from urinary bladder cancer in Italy.

This paper examines changes in mortality from urinary bladder cancer in Italy during the years 1950-81 in relation to changes in smoking habits and in coffee, cocoa and tea consumption. The authors found that, in both sexes and for all ages, mortality has been increasing throughout this period, although the rates and relative increase have consistently been much lower and more gradual for women than for men. From the analysis of age-cohort-period variation it seems that bladder cancer mortality can also be influenced by changes in smoking habit patterns and by variation in the quality of diagnoses. The increase in death risk for both sexes in Italy up to cohorts born around 1905-10 can be related to occupational exposures and to high-tar-content tobacco smoking, whereas the tendency for mortality rates to stabilize and decline in cohorts born after 1910 were influenced by changes from high-tar-content and no-filter cigarettes to low-tar-content and filter cigarettes and by prevention measures taken in at-risk working environments. The consumption of coffee, cocoa and tea does not seem to be related to the increase in bladder cancer risk in Italy.