The long-term effect on surgery-free survival of biological compared to conventional therapy in Crohn's disease in real world-data: a retrospective study.

BACKGROUND: The introduction of biological drugs has led to great expectations and growing optimism in the possibility that this new therapeutic strategy could favourably change the natural history of Inflammatory Bowel Disease (IBD) and, in particular, that it could lead to a significant reduction in surgery in the short and long term. This study aims to assess the impact of biological versus conventional therapy on surgery-free survival time (from the diagnosis to the first bowel resection) and on the overall risk of surgery in patients with Crohn's disease (CD) who were never with the surgical option. METHODS: This is a retrospective, double-arm study including CD patients treated with either biological or conventional therapy (mesalamine, immunomodulators, antibiotics, or steroids). All CD patients admitted at the GI Unit of the S. Salvatore Hospital (L'Aquila. Italy) and treated with biological therapy since 1998 were included in the biological arm. Data concerning the CD patients receiving a conventional therapy were retrospectively collected from our database. These patients were divided into a pre-1998 and post-1998 group. Our primary outcome was the evaluation of the surgery-free survival since CD diagnosis to the first bowel resection. Surgery-free time and event incidence rates were calculated and compared among all groups, both in the original population and in the propensity-matched population. RESULTS: Two hundred three CD patients (49 biological, 93 conventional post-1998, 61 conventional pre-1998) were included in the study. Kaplan-Meier survivorship estimate shows that patients in the biological arm had a longer surgery-free survival compared to those in the conventional arm (p = 0.03). However, after propensity matching analysis, conducted on 143 patients, no significant difference was found in surgery-free survival (p = 0.3). A sub-group analysis showed shorter surgery-free survival in patients on conventional therapy in the pre-biologic era only (p = 0.02; Hazard Ratio 2.9; CI 1.01-8.54) while no significant difference was found between the biologic and conventional post-biologic groups (p = 0.15; Hazard Ratio 2.1; CI 0.69-6.44). CONCLUSION: This study shows that the introduction of biological therapy has only a slight impact on the eventual occurrence of surgery in CD patients over a long observation period. Nevertheless, biological therapy appears to delay the first intestinal resection.

From the bench to practice - Field integration of community-based services for older citizens with different levels of functional limitation across European Regions.

The meeting of the European Innovation Partnership on Active and Healthy Ageing (EIPonAHA) action group A3 together with members of the Reference site collaborative network (RSCN) in December 2019 in Rome focused on integration of evidence-based approaches on health and care delivery for older citizens at different levels of needs with expertise coming from stakeholder across Europe. It was the final aim of the group to co-create culturally sensitive pathways and facilitate co-ownership for further implementation of the pathways in different care systems across Europe. The study design is a mixed method approach. Based on data analysis from a cohort of community-dwelling over-65 citizens in the framework of a longitudinal observational study in Rome, which included health, social and functional capacity data, three personas profiles were developed: the pre-frail, the frail and the very frail personas. Based on these data, experts were asked to co-create care pathways due to evidence and eminence during a workshop and included into a final report. All working groups agreed on a common understanding that integration of care means person-centered integration of health and social care, longitudinally provided across primary and secondary health care including citizens' individual social, economic and human resources. Elements for consideration during care for pre-frail people are loneliness and social isolation, which, lead to limitation of physical autonomy in the light of reduced access to social support. Frail people need adaption of environmental structures and, again, social resource allocation to maintain at home. Very frail are generally vulnerable patients with complex needs. Most of them remain at home because of a strong individual social support and integrated health care delivery. The approach described in this publication may represent a first approach to scaling-up care delivery in a person-centered approach.

Minimally invasive right colectomy: challenges and clinical practice.

AIM: Since the introduction of laparoscopic and robotic technology in surgical practice, there have been multiple reports and a few clinical trials on their use in colorectal surgery. Although the application of laparoscopy to right colectomy has been increasingly adopted in many institutions around the world, there are still several open issues regarding the effective role of robotics and single incision surgery. This is a review of the relevant surgical literature evaluating the risks, benefits, and costs of minimally invasive approaches to right colectomy (RC) surgery. METHODS: Retrospective and prospective articles spanning the past 20 years were reviewed to identify the current application of minimally invasive surgery in RC. A review of the most relevant papers comparing open vs. laparoscopic vs. robotic approaches will illustrate the role of minimally invasive surgery in current clinical practice in terms of surgical outcomes, technical advantages and oncological outcomes. We then pooled the evidence for and against the application of laparoscopy and robotics in intracorporeal vs. extracorporeal anastomosis creation, single incision and natural orifice surgery. RESULTS: Evidence shows that compared to open surgery, laparoscopic RC provides lower postoperative morbidity, faster return to normal bowel function and a shorter length of hospital stay, with a similar oncological outcome. The application of robotics to RC procedure has proven to be safe and feasible, however the intraoperative and postoperative outcomes are similar with the laparoscopic technique and no clear advantages have been demonstrated. When adopted in a single incision technique and natural orifice surgery, robotics can help to overcome the limitations of laparoscopy, enabling the surgeon to perform scar-less surgery. CONCLUSION: Laparoscopy surgery, whenever performed by adequately trained surgeons, can be safely applied to right colectomy and should be considered as the gold standard procedure. In terms of robotic surgery, to date, this technology needs more evidence from multicenter randomized clinical trials. New tools and instruments are needed to expand the field of single incision and natural orifice surgery, and make it available in current clinical practice.

Progesterone receptor expression and proliferative activity in uterine tumours of pet rabbits.

Endometrial adenocarcinoma is the most common uterine tumour of domestic rabbits. The present immunohistochemical study examined the expression of cytokeratin 19 (CK19), the progesterone receptor (PR), the proliferation-associated antigen Ki-67 and telomerase in normal rabbit uterine tissue and examples of endometrial hyperplasia, adenoma and adenocarcinoma. Tubulopapillary adenomas and adenocarcinomas were the most common histological subtypes in this series. Cytoplasmic expression of CK19 was recorded in two of three samples of normal endometrium and in one of three samples of endometrial hyperplasia, in all adenomas and five of six adenocarcinomas. PR was expressed within the nucleus of normal endometrial cells and in one of three samples of endometrial hyperplasia, each of four adenomas and in four of six adenocarcinomas. This finding suggests that PR expression is not directly involved in neoplastic transformation of the endometrium and that such expression is not a prognostic indicator. Nuclear labelling of telomerase activity was found in one of three normal uteri, all samples of endometrial hyperplasia, two of four adenomas, but none of the adenocarcinomas. The proliferation index as determined by Ki-67 expression was 9.7+/-2.75% (mean+/- standard-deviation (SD)) for normal endometrium, 11.29+/-2.5% for hyperplastic endometrium, 19.40+/-3.01% for benign tumours and 19.41+/-7.9% for malignant tumours. These findings may be interpreted to suggest that hormonal and anti-proliferative treatment may be more appropriate for the management of uterine carcinomas in rabbits than anti-telomerase treatment.

Pathological and molecular characteristics distinguishing contralateral metastatic from new primary breast cancer.

BACKGROUND: Breast cancer patients have a cumulative lifetime risk of 2%-15% of developing a contralateral metastatic or ex novo primary cancer. From prognostic and therapeutic viewpoints, it is important to differentiate metastatic from second primary. To distinguish these entities, we investigated whether the pattern of X chromosome inactivation could determine whether the two tumors derived from different progenitor cells. MATERIALS AND METHODS: The clonality of bilateral breast cancer was evaluated through the X-inactivation analysis using the human androgen receptor gene (HUMARA) polymorphism and the histopathologic and molecular results were compared. A different or an identical pattern of X inactivation was considered as indicator of a second primary cancer or not informative, respectively. We considered morphological indicators of a new primary cancer the absence of concordance in the histological type or a better histological differentiation. RESULTS: Ten patients with bilateral breast cancer were evaluated. Morphological criteria indicated that eight were second primary, a conclusion confirmed by the X-inactivation analysis. Two cases classified as recurrence according to morphological criteria were classified as second tumor by molecular analysis. CONCLUSION: Our results show that the HUMARA clonality assay can improve the histological parameters in differentiating metastatic cancer from second primary cancer.

Hyperhomocysteinemia is associated with cognitive impairment in multiple sclerosis.

Hyperhomocysteinemia (HHcy) has been associated with cognitive impairment in various neurological diseases. Cognitive impairment occurs early in multiple sclerosis (MS). Conflicting data have been reported regarding plasma total homocysteine (tHcy) levels in MS patients, and the impact of HHcy on cognitive impairment in MS is not known. This study investigated whether plasma total homocysteine levels are increased in MS and if HHcy is associated with cognitive impairment in MS. We compared tHcy levels in 94 patients with MS and 53 healthy age-matched controls. We used a neuropsychological test battery that included the Raven's Coloured Progressive Matrices, the Visual Search Test, the Trail Making Test A and B, the Immediate and Delayed Recall of a Short Story, the 30 Paired Word Associates, the Rey-Osterrieth Complex Figure Test, and the Semantic and Verbal Fluency Tests. Clinical (sex, age, type of MS, relapse, disease duration, coexisting disease, smoking habit, and physical disability) and laboratory variables (HHcy, low serum levels of folate and vit.B12, MTHFR genotype) were evaluated for their ability to predict cognitive impairment. The mean tHcy was higher in patients (13.19 micromol/L, SD5.58) than in controls (9.81 micromol/L, SD2.53; p < 0.001). Univariate analysis determined the following factors to be associated with cognitive impairment: higher age at observation, chronic progressive course of disease, longer disease duration,moderate or severe physical disability, and frequency of HHcy. With multivariate regression analysis, there remained a significant association only between frequency of HHcy and cognitive impairment (beta 0.262, p = 0.01). We conclude that tHcy levels are increased in MS and that HHcy is associated with cognitive impairment in this disease.

FMECA methodology applied to two pathways in an orthopaedic hospital in Milan.

INTRODUCTION: Adverse events pose a challenge to medical management: they can produce mild or transient disabilities or lead to permanent disabilities or even death; preventable adverse events result from error or equipment failure. METHODS: IRCCS Istituto Ortopedico Galeazzi implemented a clinical risk management program in order to study the epidemiology of adverse events and to improve new pathways for preventing clinical errors: a risk management FMECA-FMEA pro-active analysis was applied either to an existing clinical support pathway or to a new process before its implementation. RESULTS: The application of FMEA-FMECA allowed the clinical risk unit of our hospital to undertake corrective actions in order to reduce the adverse events and errors on high-risk procedure used inside the hospitals.

Autoimmune myocarditis and dilated cardiomyopathy: focus on cardiac autoantibodies.

Criteria of organ-specific autoimmunity are fulfilled in a subset of patients with myocarditis/dilated cardiomyopathy (DCM). In particular, circulating heart-reactive autoantibodies are found in patients and symptom-free relatives. These autoantibodies are directed against multiple antigens, some of which are expressed in the heart (organ-specific), others in heart and some skeletal muscle fibres (partially-heart specific) or in heart and skeletal muscle (muscle-specific). Distinct autoantibodies have different frequency in disease and normal controls. Different techniques detect one or more antibodies, thus they cannot be used interchangeably for screening. It is unknown whether the same patients produce more antibodies or different patient groups develop autoimmunity to distinct antigens. IgG antibodies, shown to be cardiac and disease-specific for myocarditis/DCM, can be used as autoimmune markers for identifying patients in whom immunosuppression may be beneficial and relatives at risk. Some autoantibodies may also have a functional role, but further work is needed.

Ki-ras activation in vitro affects G1 and G2M cell-cycle transit times and apoptosis.

Mutant ras genes occur frequently in human neoplasia and, in particular, in pancreatic, colorectal, and lung adenocarcinomas. Recent evidence suggests that G-->T and G-->C transversions of the Ki-ras gene in codon 12 may lead to biological effects in vitro and in vivo that may be associated with an abnormal cell cycle and increased tumour aggressiveness. The role of Ki-ras activation (a G-->C transversion in codon 12, arginine for glycine) in the cell cycle and apoptosis was investigated using control and permanently transfected NIH3T3 mouse fibroblasts. Flow cytometry was used to evaluate the G1-, S- and G2M-phase transit times, the potential doubling time, the growth fraction, and the cell loss factor during asynchronous exponential growth. Apoptosis was induced in both cell lines by absence of growth factors for an extended period of time (72 h) and quantitatively evaluated using the TUNEL method coupled with flow cytometry. It was found that codon 12 G-->C Ki-ras transfected cells compared with controls, had a significant prolongation of G1 by about 50%, a reduction of the G2M transit time by 30%, and a decrease of the cell loss factor by about 90%. Apoptotic cells were about 10% in control and less than 0.5% in Ki-ras transfected cells after 72 h starvation-confluency. These data suggest that codon 12 G-->C Ki-ras activation in mouse NIH3T3 fibroblasts is associated with deregulation of checkpoint controls in the G1 and G2M phases of the cell cycle and inhibition of apoptosis. It appears plausible that these cell mechanisms are related to a proliferative advantage and that they may also be important in the progression of human tumours characterized by specific Ki-ras mutations.

Analysis of erythrocyte glycophorin-A variants by flow cytometry in lung disease patients detects the effect of tobacco smoke.

The glycophoryn A (GPA) assay evaluates somatic in vivo mutations. It is considered a cumulative biodosimeter for genotoxic exposures and is under evaluation in cancer risk assessment. GPA, a polymorphic membrane protein of the erythrocytes, determines the MN blood groups. The NO and NN variant frequencies (VF) may be detected in MN subjects (about 50% of the population) by flow cytometry using two differently labelled antibodies. We explored if GPA NO and NN VF might be relevant to the assessment of individual lung cancer risk and susceptibility, in a small population with a high prevalence of heavy tobacco smokers: 8 lung cancer patients and 16 subjects with non-malignant lung diseases associated with increased risk of lung cancer. There was a wide interindividual variability and complete overlap between non-neoplastic and neoplastic patients. A significant positive correlation was seen with smoking duration in NO VF (p = 0.04, age-adjusted). Current smokers (n = 12) displayed higher NO values than never (n = 1) or ex-smokers (n = 11), 36.3 +/- 18.2 and 21.0 +/- 13.2, respectively (p < 0.01). No association was shown with occupational exposure. The present exploratory study suggests that assessment of individual lung cancer risk and susceptibility by the GPA assay does not seem to be feasible. The assay appears to provide a biomarker of longterm exposure to tobacco smoke.

Cell cycle variations in azoxymethane-induced rat colorectal carcinogenesis studied by flow cytometry.

Cell cycle variations and DNA aneuploidy, were investigated in different phases of azoxymethane (AOM)-induced colon carcinogenesis in rats by flow cytometry. K-ras gene mutations (transitions Gright curved arrow A) were frequently detected in aberrant crypt foci (ACF) initial pre-neoplastic lesions. The fraction of cells in the G2M-phase of the cell cycle was higher in ACF compared to the normal mucosa of control rats. A similar modification of the cell cycle was found in adenomas and adenocarcinomas but, unexpectedly, also in morphologically normal mucosa from AOM-treated animals indicating that AOM treatment permanently modifies cell cycle control in rat colon mucosa. These alterations, however, were not associated with DNA aneuploidy as reported in human sporadic colorectal cancer, suggesting that tumour development in AOM-treated rats is less dependent on aneuploidy.

Intratumor heterogeneity of chromosome 1, 7, 17, and 18 aneusomies obtained by FISH and association with flow cytometric DNA index in human colorectal adenocarcinomas.

BACKGROUND: The origin and evolution of somatic chromosome aberrations in colorectal cancer is still poorly understood. The data in the literature suggest that some specific chromosome aberrations are more common. It is not known, however, if there is a correlation of these with near-diploid and high aneuploidy previously proposed to be a characteristic of the adenoma-carcinoma sequence. METHODS: Chromosome 1, 7, 17 and 18 numerical aberrations and 1p deletions were evaluated by fluorescence in situ hybridization analysis for 20 human sporadic colorectal adenocarcinomas in 70 distinct tumor sectors and correlated with flow cytometric DNA index (DI) values. RESULTS: Aneusomy for at least one of the investigated chromosomes was observed in 60 of 70 tumor sectors corresponding to 19 of 20 adenocarcinomas (95%). Deletions at 1p, observed in 8 of 18 adenocarcinomas (44%), were intratumor homogeneous in 7 of 8 tumors. In contrast, the other aberrations were intratumor heterogeneous. Aneusomies of chromosomes 1, 7, and 17 were strongly associated with DNA high aneuploidy (DI > or = 1.4), whereas aneusomy of chromosome 18 and 1p deletions were equally common among DNA diploid and near-diploid tumors (DI < 1.4 and DI not equal to 1). CONCLUSIONS: Overall, these data suggest the existence of different aneuploidization routes correlated with specific chromosome aberrations. In addition, intratumor homogeneity of 1p deletions appears to be an indication of early occurrence or strong selection. We also suggest that tumors with monosomies and in particular monosomies-trisomies for the same chromosomes support a model of aneuploidization and chromosome instability during the colorectal tumor progression based on loss of symmetry during chromosome segregation (Giaretti: Lab Invest 71:904-910, 1994).

Epoetin alfa in low hematocrit patients to facilitate autologous blood donation in total hip replacement: a randomized, double-blind, placebo-controlled, dose-ranging study.

We investigated the safety and efficacy of preoperative epoetin alfa used in conjunction with preoperative autologous blood donation (PAD) in 40 anemic orthopedic surgical patients undergoing hip replacement surgery [hematocrit (Hct)

Intratumor distribution of 1p deletions in human colorectal adenocarcinoma is commonly homogeneous: indirect evidence of early involvement in colorectal tumorigenesis.

BACKGROUND: Cytogenetics and molecular biology studies have indicated that a large subset of human colorectal adenocarcinomas have distal 1p chromosome arm deletions. The aim of this study was to evaluate the intratumor distribution of 1p deletions under the assumption that homogeneity is an indication of early occurrence. METHODS: Seventy-nine histologically selected primary sectors (40 superficial and 39 deep) and 3 lymph node metastases obtained from 20 human sporadic adenocarcinomas were analyzed. Interphase two-color fluorescence in situ hybridization (FISH) was applied to cytocentrifuged nuclei using a centromeric probe for chromosome 1 and a telomeric probe mapping to the 1p36 band. RESULTS: Deletions at 1p were observed in 35 of 82 tumor samples corresponding to 9 of 20 adenocarcinomas analyzed (45%). Seven of the 9 adenocarcinomas with 1p deletions showed an intratumor presence of these aberrations in all the different tumor sectors. CONCLUSIONS: These data, acquired by FISH interphase cytogenetics, confirm that 1p deletions in colorectal adenocarcinoma are common and suggest that this structural chromosomal aberration occurs mainly as an early event in colorectal tumorigenesis.

Correlation between 1p deletions and aneusomy in human colorectal adenomas.

Human sporadic colorectal adenomas are characterized by a relatively high occurrence of aneuploidy. Similarly, 1p deletions have been reported to be an early event in colorectal tumorigenesis, while chromosome 7, 17 and 18 gain/losses were also found. The present study investigated 1p deletions, the numerical aberrations of chromosomes 1, 7, 17 and 18, and the nuclear DNA content as obtained by flow cytometry in a series of 34 human sporadic colorectal adenomas. From these adenomas, 51 intra-adenoma regions were microdissected according to 2 degrees of dysplasia and presence of foci of early cancer. Isolated epithelial nuclei were analyzed by fluorescence in situ hybridization interphase cytogenetics using centromeric probes for chromosomes 7, 17 and 18 and, in a double-target analysis, a centromeric probe for chromosome 1 simultaneously with a telomeric probe mapping to the 1p36 band. Aneuploidy incidence due to presence of numerical aberrations for at least one among the investigated chromosomes and/or abnormal flow-cytometric DNA content was 35%, while 1p deletion incidence was 38%. The correlation of 1p deletions with aneuploidy was statistically highly significant (p = 0.003), suggesting that loss of genes in this region may be implicated in chromosome instability.

Preliminary characterization of a monoclonal antibody (AS-2) against cell cycle related proteins.

A monoclonal antibody (AS-2) raised by using isolated nuclei from a human erythroleukemia cell line as immunogen is described. AS-2 was of IgM type and recognized proteins present in both isolated cytoplasms and nuclei. The molecular weight of the AS-2 recognized proteins in the cytoplasm was 200 kDa and 70 and 60 kDa in the nucleus. The relative amount of these proteins were measured simultaneously with DNA content by flow cytometry. We found the highest protein content (or stainability) for both cells and nuclei in late-G1, S and G2, at approximately the same level, and the lowest content in M and early-G1. Sorting based on DNA content and AS-2 associated fluorescence helped identifying the staining pattern of cells and nuclei. Interphase isolated nuclei and cell cytoplasms were characterized by interdispersed staining over the entire surfaces while mitoses showed two dots only. The present preliminary data indicate that the proteins recognized by the AS-2 monoclonal are cell cycle related and suggest that in mitoses they are associated with the centrosomes.

[Atherosclerosis in childhood: the role of obesity]. / Aterosclerosi in età pediatrica: ruolo dell'obesità.

A variety of studies indicates that the process of atherosclerosis begins in childhood and progresses during adulthood. Chronic obesity, inadequate caloric intake, and hypertension and smoke, are associated with an increased cardiovascular disease. The aim of this study is to investigate if the presence of some risk factors during adolescence may involve in accelerated atherosclerosis disease. 50 subjects, median age 11 +/- 0.6 SD (27 females, and 23 males) are admitted to the study. After overnight fasting we have investigated: lipoproteina A (nephelometric test), glycemia and insulin baseline and after load 120', tryglycerides, cholesterolo, apolipoproteina A, B, plasma concentrations. In addition to general medical evaluation, anthropometric measurements of weight, height, blood pressure, BMI, overnight ratio were calculated according to Tanner's charts. The means anthropometric and metabolic values in different groups were compared. One group affected with abdominal obesity state (waist-hips ratio > 0.9), the second with mid obesity condition (waist-hip ratio < 0.9). Tryglycerides, cholesterolo, insulin plasma concentrations in both groups were considered similar. However in the first group higher levels of apolipoproteina A (means 102 + 10.2 SD) and lipoproteina A were demonstrated (P = 0.03 in males, P = 0.01 Statview for Mann Whitney test). Childhood is an important period for the development of the atherosclerosis such as the presence of obesity during this time has a very high likelihood of persisting into adulthood. The severity of obesity in adults is greater in those who were obese as adolescents. In accord with other authors we have not observed abnormal tryglicerides and cholesterolo plasma concentrations, which probably are found in adulthood obesity. We believe indeed the risk factors are different in obesity of childhood, atherosclerosis may be induced by high endogenous insulin secretion and abnormal uptake of lipoprotein. However the potential consequence of excessive insulin secretion could be due in part to insulin effects on recruitment of histiocytosis cells during the development of atheroma and through the modulation of hepatic production and peripheral uptake of lipoproteins.

Evidence of apoptosis in neuroblastoma at onset and relapse. An analysis of a large series of tumors.

Neuroblastoma (NB) is a tumor of pediatric age that is associated with high mortality in metastatic stages, although stage IVS patients undergo frequent spontaneous regression. Since apoptosis has been proposed as a possible cause of remission among cancer patients, we tested this hypothesis among both localized and metastatic NB and, in particular, NB metastatic stage IVS. We have assayed 36 localized and 117 metastatic neuroblastomas for evidence of internucleosomal DNA degradation and confirmed DNA fragmentation by the flow cytometric Terminal deoxynucleotidyl Transferase method, which also allowed us to measure DNA content and cell cycle phases. These techniques provided evidence of apoptosis in 18 out of 153 samples (11.8%), that were equally distributed among all stages except IVS, i.e. 11.1% in stage I (2/18), 11.1% in stage II (2/18), 13.2% in stage III (5/38), 13.4% in stage IV (9/67), and 0% in stage IVS (0/12). Tumor tissue samples collected at onset and also at relapse for the same patients showed that apoptosis may occur at relapse. In addition, cells appear to undergo apoptosis independently from N-myc amplification, cell cycle phase and DNA ploidy. In conclusion, apoptosis seems to take place with about an equal frequency for both favourable and unfavourable stages with an exception for IVS. Since DNA fragmentation remained undetected in stage IVS, we suggest that apoptosis is not a mechanism of spontaneous regression for these patients. A better basic understanding of the complex molecular mechanisms and biochemical pathways that control apoptosis in neuroblastoma appears to be necessary.