PKCε is a negative regulator of PVAT-derived vessel formation.

RATIONALE: Vessel formation is a crucial event in tissue repair after injury. Thus, one assumption of innovative therapeutic approaches is the understanding of its molecular mechanisms. Notwithstanding our knowledge of the role of Protein Kinase C epsilon (PKCε) in cardio-protection and vascular restenosis, its role in vessel progenitor differentiation remains elusive. OBJECTIVE: Given the availability of PKCε pharmacological modulators already tested in clinical trials, the specific aim of this study is to unravel the role of PKCε in vessel progenitor differentiation, with implications in vascular pathology and vasculogenesis. METHODS AND RESULTS: Mouse Peri-Vascular Adipose Tissue (PVAT) was used as source of mesenchymal vessel progenitors. VEGF-induced differentiation of PVAT cells down-regulates both PKCε and p-PAK1 protein expression levels. PKCε overexpression and activation: i) reduced the expression levels of SMA and PECAM in endothelial differentiation of PVAT cells; ii) completely abrogated tubules formation in collagen gel assays; iii) increased the expression of p-PAK1. CONCLUSION: PKCε negatively interferes with vessel progenitor differentiation via interaction with PAK-1.

Gene expression profiling reveals GC and CEACAM1 as new tools in the diagnosis of lung carcinoids.

BACKGROUND: Classification of lung carcinoids into typical and atypical is a diagnostic challenge since no immunohistochemical tools are available to support pathologists in distinguishing between the two subtypes. A differential diagnosis is essential for clinicians to correctly discuss therapy, prognosis and follow-up with patients. Indeed, the distinction between the two typical and atypical subtypes on biopsies/cytological specimens is still unfeasible and sometimes limited also after radical surgeries. By comparing the gene expression profile of typical (TC) and atypical carcinoids (AC), we intended to find genes specifically expressed in one of the two subtypes that could be used as diagnostic markers. METHODS: Expression profiling, with Affymetrix arrays, was performed on six typical and seven atypical samples. Data were validated on an independent cohort of 29 tumours, by means of quantitative PCR and immunohistochemistry (IHC). RESULTS: High-throughput gene expression profiling was successfully used to identify a gene signature specific for atypical lung carcinoids. Among the 273 upregulated genes in the atypical vs typical subtype, GC (vitamin D-binding protein) and CEACAM1 (carcinoembryonic antigen family member) emerged as potent diagnostic markers. Quantitative PCR and IHC on a validation set of 17 ACs and 12 TCs confirmed their reproducibility and feasibility. CONCLUSIONS: GC and CEACAM1 can distinguish between TC and AC, defining an IHC assay potentially useful for routine cytological and histochemical diagnostic procedures. The high sensitivity and reproducibility of this new diagnostic algorithm strongly support a further validation on a wider sample size.

Assessment of biophysical therapy in the management of pain in current medical practice compared with ibuprofen and placebo: a pilot study.

Pain management is a daily part of current medical practice. The aim of this pilot study was to assess the efficacy of a biophysical procedure (Med Select 729) compared to a usual pain killer drug (Ibuprofen), and to placebo in order to disclose some effective procedures to be employed especially in elderly people with multiple comorbidities, in patients with allergy to chemical drugs or previous side effects, in non-responders to usual medications, and in chronic diseases to reduce overload. A total of 66 patients were divided in 3 groups. After one week of biophysical therapy they showed similar effect to ibuprofen and after one month the statistical significance was achieved with p less than 0.02 in comparison to placebo. We conclude that biophysical therapy was shown to be an effective and safe procedure for the management of pain in current medical practice.

Immunologic follow-up of infants treated with granulocyte transfusion for neonatal sepsis.

The immunologic status and the occurrence of alloimmunization against granulocytes, platelets, lymphocytes, and red cells was evaluated in 33 babies who received granulocyte transfusion because of neonatal sepsis. Nine age-matched babies were examined as control. A first group of 19 infants was examined only once between 6 and 23 months of age. Alloantibodies were searched by the following serologic methods: standard techniques for red cell antibodies; lymphocytotoxicity test; agglutination and immunofluorescence tests on granulocytes and platelets. No antibodies were demonstrated. The immunologic profile was investigated by determining the Ig levels, the percentage of E rosette-forming cells, and the lymphocyte blastic response to phytohemagglutinin and concanavalin A. Granulocyte function was studied by phagocytosis and killing of Candida. No significant differences were observed between treated and control babies. In a second group of 14 infants the occurrence of early immunization within 3 to 9 weeks after the last transfusion was investigated. No evidence of early immunization was found. The present data suggest that following neonatal granulocyte transfusion the risk of adverse immune reactions should be low.

Ureaplasma urealyticum and pulmonary outcome in a neonatal intensive care population.

OBJECTIVE: To determine whether the presence of Ureaplasma urealyticum in the respiratory tracts of very low birth weight infants is associated with increased risk of pneumonia, radiographic evidence of severe bronchopulmonary dysplasia during the second or third week of life (precocious) and oxygen dependency at 36 weeks of corrected postnatal gestational age. METHODS: From October, 1993, to January, 1996, all infants who met the following entry criteria were enrolled in the study: birth weights < or = 1500 g; admission to the NICU within 24 h after birth; evidence on admission of respiratory distress; and no prior antibiotic treatment. Infants were cultured for mycoplasmas, viruses, chlamydiae and aerobic and anaeroic bacteria. RESULTS: Ninety-four critically ill newborns constituted our study cohort. Within 7 days of birth more infants with U. urealyticum infection showed radiographic features of pneumonia (53.1%, 25 of 47) than infants without U. urealyticum infection (21.2%, 10 of 47). Infants with U. urealyticum were more likely to have radiographic evidence of precocious bronchopulmonary dysplasia than those without this isolate (22.5%, 9 of 40 vs. 2.3%, 1 of 42). The relative risk of oxygen dependency at 36 weeks of corrected age in U. urealyticum-positive infants was 11.0 times that in U. urealyticum-negative infants (95% confidence interval, 1.6 to 75.5). Association of U. urealyticum and chronic lung disease at this age was not weakened after adjustments were made in a multivariate analysis for other significant risk factors including gestational age and presence of a patent ductus arteriosus. CONCLUSIONS: Isolation of U. urealyticum from respiratory tracts is associated with radiographic changes compatible with pneumonia within 7 days of birth, precocious bronchopulmonary dysplasia and severe pulmonary outcome.

Comparing the effects of nasal synchronized intermittent positive pressure ventilation (nSIPPV) and nasal continuous positive airway pressure (nCPAP) after extubation in very low birth weight infants.

In this study we hypothesized that nasal synchronized intermittent positive pressure ventilation (nSIPPV) would provide more ventilatory support than nasal continuous positive airway pressure (nCPAP) in the immediate post-extubation period in very low birth weight (VLBW) infants. We tested this hypothesis by comparing the effects of these two ventilatory techniques on ventilation, gas exchange, and patient inspiratory effort in 11 preterm infants immediately after extubation. All neonates studied (BW: 1141+/-(SEM) 53 g; GA: 28.1+/-(SEM) 0.5 wks) had received mechanical ventilation because of respiratory distress at birth and were extubated by day 14 of life. Nasal SIPPV and nCPAP were applied in random order to each infant after extubation so that each was his/her own control. Both nCPAP and nSIPPV were delivered at end-expiratory pressures (PEEP) of 3 cm H2O. Inspiratory times (Ti) and peak inspiratory pressures set during nSIPPV were the same as those used at the time of extubation. Recordings lasted 45 min in each mode of ventilation. Tidal volume (Vt), minute volume (Ve), respiratory rate (RR), airway pressure (Paw), transcutaneous PO2 (TcPO2) and PCO2 (TcPCO2) as well as phasic esophageal pressure deflections (Pe), as an estimate of inspiratory effort, were measured. The measurements obtained during both modes of ventilation indicated significant differences between the two techniques. Indeed, application of nSIPPV was associated with a statistically significant increase in Vt and Ve. In addition, Pe decreased by 30% during nSIPPV (P<0.01). TcPCO2 was statistically significantly lower during nSIPPV than nCPAP, and RR as well. These data therefore suggest that nSIPPV may provide more ventilatory support than nCPAP in the post-extubation period with less patient inspiratory effort.

Topography and ultrastructure of kidney lymphatics in some hibernating bats.

The kidney lymphatic system of some bats consists of intraparenchymal (interlobar, arcuate, and interlobular) and extraparenchymal vessels (capsular and prehilar connective). These vessels drain lymph via precollecting and prenodal collecting lymphatics into a hilar lymph node. There are no lymphatics in the renal medulla. The lymphatic vasculature (precollecting vessels excluded) is characterized by an endothelial wall lacking basal lamina and fenestrations. The endothelial cells, mostly rectangular in shape, are joined together by overlapping, end-to-end, and complex interdigitating junctions. Cytoplasmic expansions profile and thickness, intercellular junctions and particularly the different categories of uncoated vesicles (free or opened on luminal or abluminal surface) show qualitative and quantitative seasonal variations. Luminal and abluminal cytoplasmic processes appear (when analyzed in tridimensional reconstructions) as "intraendothelial channels." The increased number of these structures during summer characterizes them as dynamic elements and supports the concept of an active role played by them in transendothelial transport. Nevertheless, the main functional role is still ascribed (in addition to membrane transport mechanisms) to the vesicular system, also defined as the "vesicular route." We did not find any open intercellular junctions.

Integrating content-based retrieval in a medical image reference database.

Image reference databases (IRDBs) are a recurrent research topic in medical imaging. Most IRDBs are designed to help experienced physicians in diagnostic tasks and require that users have prior extensive knowledge of the field for their use to be fruitful. Therefore, the educational potential of such image collections cannot be exploited thoroughly. In this paper we propose an image-indexing method to extend the functionalities of an existing medical IRDB and allow for its use in educational applications, as well as in computer-assisted diagnosis. Our method, based on the Kahrunen-Leève transform, has been used to develop a content-based search engine for tomographic image databases on which we are presently experimenting and which we aim to integrate into a working radiological IRDB installed at the University of Florence. Results achieved in our preliminary tests are also reported.

Fine structure of bat deep posterior lingual glands (von Ebner's)

We studied the morphology and ultrastructure of the bat (Pipistrellus k.k. and Rhinolophus f.e.) deep posterior lingual glands (Ebner's glands) during hibernation, summer and after stimulation with pilocarpine. Ebner's glands are formed by serous tubulo-alveolar adenomeres and by an excretory system organized in intercalated ducts, long excretory ducts and a main excretory duct. The latter opens in the vallum which surrounds the circumvallate papillae and in the groove of the foliate papillae. The secretory cells, which lack basal folds, show abundant and dense granules (PAS+, Alcian blue -), microvilli (scarce during hibernation), a Golgi apparatus (well developed during summer and after stimulation with pilocarpine), a large nucleus and RER cisternae stacked at the basal pole. Centrioles, lipid droplets, heterogeneous bodies (in content and density, probably lipofuscin bodies), lysosomal multivesicular bodies and large, dense granules with a microcrystalline structure were also encountered. The lateral membranes of adjacent cells are joined by desmosomes; their interdigitations are neither numerous nor prominent during summer. Microfilaments, often gathered in small bundles, lie in the lateral, peripheral cytoplasm without any relation with desmosomes. In summer and particularly after stimulation with pilocarpine, the apical pole of the secretory cells is characterized by many long microvilli, pedunculated hyaloplasmic protrusions and secretory granules. During hibernation the lumen is filled with secretory material. Myoepithelial cells are arranged among secretory cells or between them and the basal lamina. The short intercalated ducts show similarities with the analogous ducts of the parotid gland. Striated ducts are absent. Excretory ducts are endowed with: a) an inner layer of cuboidal cells characterized by poorly developed cytoplasmic organelles, rare dense granules and a few small microvilli; b) an outer layer of basal cells lying on the basal lamina. Myoepithelial cells are absent. The main excretory duct is lined by a stratified epithelium with an inner layer of conical-pyramidal cells surrounded by two-three rows of basal cells. The conical-pyramidal cells show poorly developed organelles, an apical border with small short microvilli and a prominent terminal web.

Seasonal changes of parafollicular and follicular cells of the dormouse thyroid (Myoxus glis): an ultrastructural and immunocytochemical study.

The follicular epithelium of dormouse thyroid consists of two distinct cellular types, follicular and parafollicular cells. Parafollicular cells can be easily identified by their high cytoplasmic dye-affinity for phloxine, round to ovoid shape, basal arrangement and lack of contact with follicular colloid. The wide cytoplasmic matrix is clear and contains many secretory granules of variable electron density whose contents histochemically appears to be proteic with a lean glucidic component. Furthermore immunocytochemical reactions with antibodies against calcitonin and somatostatin showed that both hormones are co-stored in the secretory granules of all parafollicular cells. Both follicular and parafollicular cells show seasonal morphological variations in their secretory activity. Follicular cell activity is high in summer, reaches a peak in late fall or prehibernation and progressively slows down throughout hibernation. Parafollicular cells exhibit a fair synthetic activity in summer, in fall, and in the animals captured during winter hibernating sleep and killed after 12 days stay in laboratory. In winter sleep, granules with interrupted membrane and cottony contents are prevalent and the ultrastructural aspects suggest an intense discharge of secretion. The results are compared with those from other hibernating mammalians and discussed in the light of blood calcium values and seasonal balances of other metabolisms.

[Current problems of the surgical treatment of rectal cancer: analysis of the literature and personal experience]. / Problematiche attuali nella terapia chirurgica del cancro del retto: analisi della letteratura ed esperienza personale.

The surgical procedure is a crucial factor in preventing local recurrence in rectal cancer, and total mesorectal excision (TME) particularly is widely accepted as being associated with a decreased local recurrence rate. In this study, concerning 187 patients with rectal cancer, we compare conventional surgery, performed in 140 patients from 1979 to 1993, with a standardised TME procedure in 47 patients over the period from 1994 to 1998. The first group not treated by TME were operated on for 56 (40%) tumours of the upper rectum and 84 (60%) of the lower rectum; 35 (25%) were Dukes' A, 77 Dukes' B and 28 (20%) Dukes' C. 42 abdominoperineal amputations (30%) and 98 anterior resections (70%) were performed. The second group in which TME was performed comprised 17 (36%) tumours of the upper rectum and 30 (64%) of the lower rectum, 8 (17%) in stage I AJCC (Dukes' A), 16 (34%) in II (B) and 23 (49%) in III (C). 9 abdominoperineal amputations (19%) and 38 anterior resections (81%) were performed, 8 (21%) with an ultra-low anastomosis. In the first group of patients we observed 28 local recurrences (20%) and a 5-year disease-free survival in 50% of cases. In the second group we achieved a decreased rate of local recurrence (10.6%) which is about half that observed after conventional surgery, but there was no significant difference in survival. These data confirm the effectiveness of TME in reducing local recurrence rate, according to the literature; in future this procedure can get to reconsider the role of adjuvant therapy in the management of rectal cancer.

[Mortality and care level of very low birth weight newborn infants. A population study]. / Mortalità e livello delle cure in neonati di peso molto basso. Uno studio di popolazione.

A population study on 314 very low birth weight infants (VLBW) was carried out in 1987 in the Lazio Region of Italy to investigate the relation between the availability at birth of neonatal intensive care and infant mortality. Fifty-two percent of VLBW infants did not survive the first year of life. The mortality Odds Ratios, adjusted for four potential confounding variables, did not show a beneficial effect of Maternity units with neonatal intensive care (level 3) compared with those with special (level 2) and normal care (level 1). The overall high crude mortality rate together with the homogeneity of odds ratios among the different levels of care suggest that, when a regionalized perinatal care system is missing, as in Lazio region, the availability of neonatal intensive care, per se, does not improve the survival on this group of infants.

Old and new diagnostic approaches for Q fever diagnosis: correlation among serological (CFT, ELISA) and molecular analyses.

The objective of this study was to evaluate the performance of the complement fixation test (CFT) with respect to ELISA for the serological diagnosis of Q fever and to assess the role of serology as a tool for the identification of the shedder status. During 2009-2010, sera from 9635 bovines and 3872 small ruminants (3057 goats and 815 sheep) were collected and analyzed with CFT and ELISA. In addition, 2256 bovine, 139 caprine and 72 ovine samples (individual and bulk tank milk samples, fetuses, vaginal swabs and placentae) were analyzed with a real-time PCR kit. The relative sensitivity (Se) and specificity (Sp) of CFT with respect to ELISA were Se 26.56% and Sp 99.71% for cattle and Se 9.96% and Sp 99.94% for small ruminants. To evaluate the correlation between serum-positive status and shedder status, the ELISA, CFT and real-time PCR results were compared. Due to the sampling method and the data storage system, the analysis of individual associations between the serological and molecular tests was possible only for some of the bovine samples. From a statistical point of view, no agreement was observed between the serological and molecular results obtained for fetus and vaginal swab samples. Slightly better agreement was observed between the serological and molecular results obtained for the individual milk samples and between the serological (at least one positive in the examined group) and molecular results for the bulk tank milk (BTM) samples. The CFT results exhibited a better correlation with the shedder status than did the ELISA results.