Human cleft lip and palate fibroblasts and normal nicotine‐treated fibroblasts show altered in vitro expressions of genes related to molecular signaling pathways and extracellular matrix metabolism

Nonsyndromic cleft lip with or without cleft palate (CLP) is a frequent craniofacial malformation caused by both genetic and environmental factors. Maternal smoking during pregnancy is a known risk factor, due to the teratogenic role of nicotine. To assess and compare the impact of CLP and nicotine, we studied the quantitative expression of genes involved in signaling pathways and extracellular matrix (ECM) metabolism in human normal nicotine‐treated (NicN) and CLP fibroblasts compared to normal control (CTRL) cells. Palatal fibroblast cultures from seven CLP children and seven age‐matched CTRL subjects were established and subconfluent cells incubated for 24 h without (CTRL and CLP fibroblasts) or with (NicN fibroblasts) 0.6 mM nicotine. Gene expressions were analyzed by real‐time quantitative PCR. For the first time, a regulated cholinergic signaling in our human fibroblasts in vitro was demonstrated. Members of TGF‐beta, retinoic acid (RA), and GABA‐ergic signaling systems were also differently regulated. Among the ECM genes, fibronectin, syndecan, integrin α2, and MMP13 genes were concordantly modulated, while integrin β5, and decorin genes were discordantly modulated. Interestingly, nicotine treatment regulated gene expressions of CD44 and CLPTM1, two candidate genes for CLP. Our findings show a positive association between nicotine treatment and CLP phenotype. Results suggest that nicotine deranges normal palate development, which might contribute to the development of a CLP malformative phenotype, through the impairment of some important signaling systems and ECM composition. J. Cell. Physiol. 222: 748–756, 2010. © 2009 Wiley‐Liss, Inc.     

A rat mammary gland cancer cell with stem cell properties of self-renewal and multi-lineage differentiation

The cancer stem cell hypothesis posits that tumors are derived from a single cancer-initiating cell with stem cell properties. The task of identifying and characterizing cancer-initiating cells with stem cell properties at the single cell level has proven technically difficult because of the scarcity of the cancer stem cells in the tissue of origin and the lack of specific markers for cancer stem cells. Here we show that a single LA7 cell, derived from rat mammary adenocarcinoma has: the ability to serially re-generate mammospheres in long-term non-adherent cultures, the differentiation potential to generate all the cell lineages of the mammary gland and branched duct-like structures that recapitulate morphologically and functionally the ductal–alveolar-like architecture of the mammary tree. The properties of self-renewal, extensive capacity for proliferation, multi-lineage differentiation and the tubular-like structure formation potential suggest that LA7 cells is a cancer stem model system to study the dynamics of tumor formation at the single cell level. Cinzia Cocola, Sveva Sanzone and Simonetta Astigiano have contributed equally to this work.     

Sulla cariologia di Gennaria diphylla (Link) Parl. (Coeloglossum diphyllum Fiori)

Abstract

OBSERVATIONS ON THE CARYOLOGY OF GENNARIA DIPHYLLA (LINK) PARL. (COELOGLOSSUM DIPHYLLUM FIORI) — Some caryological characters of Gennaria diphylla (Link) Parl, have been here investigated. Chromosome counts from somatic metaphases show a 34 chromosomes diploid set; the resting nuclei have been classified as half-reticulated with chromocenters (« noyaux semi-reticulé à chromocentres » according to Delay's classification); in prophases we have noticed that chromocenters often appear like prochromosomes. The chromosome number 17 (2n = 34) is new for the Ophrydinae.

The interest of this research lies in the fact that the species here dealt with has been differently classified by various Authors.     

Pathomechanisms of ALS8: altered autophagy and defective RNA binding protein (RBP) homeostasis due to the VAPB P56S mutation

Mutations in RNA binding proteins (RBPs) and in genes regulating autophagy are frequent causes of familial amyotrophic lateral sclerosis (fALS). The P56S mutation in vesicle-associated membrane protein-associated protein B (VAPB) leads to fALS (ALS8) and spinal muscular atrophy (SMA). While VAPB is primarily involved in the unfolded protein response (UPR), vesicular trafficking and in initial steps of the autophagy pathway, the effect of mutant P56S-VAPB on autophagy regulation in connection with RBP homeostasis has not been explored yet. Examining the muscle biopsy of our index ALS8 patient of European origin revealed globular accumulations of VAPB aggregates co-localised with autophagy markers LC3 and p62 in partially atrophic and atrophic muscle fibres. In line with this skin fibroblasts obtained from the same patient showed accumulation of P56S-VAPB aggregates together with LC3 and p62. Detailed investigations of autophagic flux in cell culture models revealed that P56S-VAPB alters both initial and late steps of the autophagy pathway. Accordingly, electron microscopy complemented with live cell imaging highlighted the impaired fusion of accumulated autophagosomes with lysosomes in cells expressing P56S-VAPB. Consistent with these observations, neuropathological studies of brain and spinal cord of P56S-VAPB transgenic mice revealed signs of neurodegeneration associated with altered protein quality control and defective autophagy. Autophagy and RBP homeostasis are interdependent, as demonstrated by the cytoplasmic mis-localisation of several RBPs including pTDP-43, FUS, Matrin 3 which often sequestered with P56S-VAPB aggregates both in cell culture and in the muscle biopsy of the ALS8 patient. Further confirming the notion that aggregation of the RBPs proceeds through the stress granule (SG) pathway, we found persistent G3BP- and TIAR1-positive SGs in P56S-VAPB expressing cells as well as in the ALS8 patient muscle biopsy. We conclude that P56S-VAPB-ALS8 involves a cohesive pathomechanism of aberrant RBP homeostasis together with dysfunctional autophagy.Subject terms: Mechanisms of disease, Amyotrophic lateral sclerosis     

Echocardiographic alterations in a child with cow's milk allergy: a case report

ABSTRACT: INTRODUCTION: Cow's milk allergy is the most frequent food allergy in Europe and western countries and shows a wide spectrum of clinical features, including atopic dermatitis and gastrointestinal disease. To the best of our knowledge, this report is the first to describe Kawasaki disease-like clinical features and echocardiographic alterations which resolved after a cow's milk-free diet. CASE PRESENTATION: We report a case of a 9-month-old Caucasian girl with atopic dermatitis who developed clinical features commonly present in Kawasaki disease (erythematous skin rash, non-exudative conjunctivitis, fissured lips and neck lymph nodes), together with mild echocardiographic alterations (perivascular brightness, pericardial effusion) in the absence of fever. These features resolved within 2 weeks after the beginning of a cow's milk-free diet. CONCLUSION: Kawasaki disease has recently been considered a possible risk factor for subsequent allergic disease secondary to immune dysfunction. This case report suggests that the immune-related alterations which are commonly present in allergic patients could be similar to the antigen-related immune response in Kawasaki disease and thus could lead to similar clinical features.     

Properdin factor B polymorphism in continental Italy and Sardinia

Summary A sample of healthy unrelated individuals (882 from Continental Italy and Sicily, and 139 from Sardinia) were typed for HLA and Bf polymorphisms. Bf gene frequencies in Continental Italy and Sicily show an intermediate position among European populations, with Lapps on one side and Basques on the other side; while a very peculiar pattern is observed among Sardinians, with an impressive similarity with French Basques and with the highest F1 gene frequency so far observed. The HLA: Bf gametic associations in Italy and Sardinia are also reported, which confirm the different ethnic background of these populations.This work was supported by Centro di Immunogenetica ed Istocompatibilità, C.N.R., Torino, Italy and by a grant of Ministero Pubblica Istruzione (M.P.I., Cap. 8551/1980).     

Bovine lactoferrin inhibits Influenza A virus induced programmed cell death in vitro

Influenza is one of the main plagues worldwide. The statistical likelihood of a new pandemic outbreak, together with the alarming emergence of influenza virus strains that are resistant to available antiviral medications, highlights the need for new antiviral drugs. Lactoferrin, a 80 kDa bi-globular iron-binding glycoprotein, is a pleiotropic factor with potent antimicrobial and immunomodulatory activities. Although the antiviral effect of lactoferrin is one of its major biological functions, the mechanism of action is still under debate. In this research, we have analyzed the effect of bovine lactoferrin (bLf) on Influenza A virus infection in vitro. Our results showed that (i) Influenza virus infected cells died as a result of apoptosis, (ii) bLf treatment inhibited programmed cell death by interfering with function of caspase 3, a major virus-induced apoptosis effector, and (iii) bLf efficiently blocked nuclear export of viral ribonucleoproteins so preventing viral assembly. These results provide further insights on the antiviral activity of bLf and suggest novel strategies for treatment of Influenza virus infection.     

Microbiological air analysis in dental surgeries: A comparison between two methods

The microbiological quality of indoor air is creating an increasing interest especially as far as places at risk such as hospitals, clinics, medical and odontological surgeries are concerned. Working with the odontologists of our province we have been carrying out a research aimed at preventing cross-infection in odontology. Data obtained from the microbiological analysis of the air in 36 surgeries using S.A.S were discussed during the V National Congress of Aerobiology. During that congress the need of setting a standardized technique of air sampling in indoor environments emerged and two routes have been identified: (1) the gravimetric technique on open plate exposed for an hour close to the dental unit and (2) the use of the volumetric sampler which gives qualitative data expressed as colonies forming units per cubic metre of air. However, both of these techniques present some problems: using the first a loss of micro-organisms has been noticed due to the variability of the air fluxes and the different weight of the biological particles; using the second one the bacterial charge is also undervalued, because of the stress suffered by the bacteria with the use of the volumetric sampler. In the light of these statements we decided to use both in dental surgeries to be able to compare the results obtained. Our project is expected to carry out at least one inspection and the relative sampling (indoor air, water of the dental unit, air of the syringe, disinfectant solution, surface tampons, biological test of sterility) in each dental surgery in the territory of our health Unit, located in Ferrara, Northern Italy.