Association between red cell distribution width and response to methotrexate in rheumatoid arthritis.

Red cell distribution width (RDW) is an unconventional biomarker of inflammation. We aimed to explore its role as a predictor of treatment response in rheumatoid arthritis (RA). Eighty-two RA patients (55 females), median age [interquartile range] 63 years [52-69], were selected by scanning the medical records of a rheumatology clinic, to analyze the associations between baseline RDW, disease activity scores and inflammatory markers, as well as the relationship between RDW changes following methotrexate (MTX) and treatment response. The lower the median baseline RDW, the greater were the chances of a positive EULAR response at three months, 13.5% [13.0-14.4] being among those with good response, vs 14.0% [13.2-14.7] and 14.2% [13.5- 16.0] (p=0.009) among those with moderate and poor response, respectively. MTX treatment was followed by a significant RDW increase (p<0.0001). The increase of RDW was greater among patients with good EULAR response, becoming progressively smaller in cases with moderate and poor response (1.0% [0.4-1.4] vs. 0.7 [0.1-2.0] vs. 0.3 [-0.1-0.8]; p=0.03). RDW is a strong predictor of early response to MTX in RA. RDW significantly increases after MTX initiation in parallel to treatment response, suggesting a role as a marker of MTX effectiveness.

Asbestos fibres detected by scanning electron microscopy in the gallbladder of patients with malignant pleural mesothelioma (MPM).

Gallbladders from patients affected by both malignant pleural mesothelioma (MPM) and important gallbladder disorders were analyzed to verify the presence of asbestos fibres. Histological thin sections were analyzed by optical microscope and variable pressure scanning electron microscopy coupled with energy dispersive spectroscopy, allowing morphological and chemical characterization of each inorganic phase observed. Fibres of chrysotile and crocidolite, minerals regulated as asbestos, were identified. By immunohistochemical analysis, connective tissue was recognized as the incorporation site. These findings confirm that asbestos fibres can reach the gallbladders of patients with MPM, for whom the development of respiratory diseases confirms asbestos exposure.

Ultrastructural detection of photosensitizing molecules by fluorescence photoconversion of diaminobenzidine.

Photodynamic therapy is a moderately invasive therapeutic procedure based on the action of photosensitizers (PSs). These compounds are able to absorb light, and dissipate energy through photochemical processes leading to the production of oxidizing chemical species (singlet oxygen, free radicals or reactive oxygen species) which can damage the cell molecular structures eventually inducing cell death. To increase the entering through the plasma membrane, a PS with suitable chemical structure can be modified by addition of chemical groups (e.g., acetate or phosphate): this affects both the fluorescence emission and of the photosensitizing properties of the native PS. The modified compounds behave as fluorogenic substrates (FSs), since inside the cell the bound groups can be enzymatically removed and the fluorescence and photosensitizing properties of the native molecules are restored. With the aim to detect the subcellular localization of photoactive molecules at transmission electron microscopy, we loaded cultured HeLa cells with two different FSs, Rose Bengal acetate (RB-Ac) or Hypocrellin B acetate (HypB-Ac), and took advantage of the photophysical properties of the intracellularly restored PS molecules to obtain the photoconversion of diaminobenzidine (DAB) into an electrondense product. We demonstrated that RB-Ac and HypB-Ac are mostly internalized by endocytosis, and are converted into the native PSs already at the cell surface. Endocytosed PS molecules apparently follow the endosomes-lysosome route, being found in endosomes, lysosomes and multivescicular bodies; PS molecules were also detected in the cytosol. This ultrastructural localization of the photoactive molecules is fully consistent with the multiorganelle photodamage observed after irradiation in culture of RB-Ac- or HypB-Ac-loaded cells. Due to the very short half-life of the oxidizing chemical species and their limited mobility, DAB deposits do localize in close proximity of the very place where photoactive molecules elicited the production of reactive oxygen species upon light irradiation. Therefore, DAB photoconversion promises to be a suitable tool for directly visualizing in single cells the PS molecules at high resolution, helping to elucidate their mode of penetration into the cell as well as their dynamic intracellular redistribution and organelle targeting.

Enigmatic question of early reactive arthritis disclosed after researches of mycoplasmas, Chlamydia trachomatis and enteropathogens following the holistic vision of human being.

An HLA-B27 genetic profile patient is fully investigated by molecular analyses after an anamnestic assessment of multi-site ecosystems, following the holistic vision of human being.VDRL and Widal-Wright (WWR) resulted positive, showing at Wright’s reaction a title of 1:40. Of all the enzymatic activities measured, only the ALP enzymatic pool activities showed a low increasing value of 297 U/L. Of all later acute phase proteins, Only C3 c protein value (127 mg/dL) and fibrinogen (376 mg/dL) were altered. Cultural and molecular oropharyngeal ecosystem investigation resulted significantly positive to Mycoplasmas(Mhand Uu) and Chlamydia trachomatis(Ct) together with a spread of saprophytic flora. From an accurate anamnesis, several and severe uro-genital clinical symptomatology emerged from birth until the beginning of rheumatologic symptomatologies that were confirmed by oldest Mh, Uu and Ctsilent chronic infections between these ecosystems. The molecular HPV research was negative, while the Thin prep pap-test was indicative of vaginosis and cellular reactive changes associated with inflammation. Parasitological research resulted positive for presence of 5-7 newly-formed G. lambliacysts for microscopic field, while digestibility test was positive for presence of several free fatty acid crystals. The remarkable presence of indigested meat fibre and several mucous dense filaments were observed. The pH value was 6.5, while blood faecal test was positive. The values observed were: ferritin 12 microg/L (10-120), total iron-binding capacity (TIBC) 310 &mgr;g/dL (300+-20), unsaturated iron-binding capacity (UIBC) 286 microg/dL (200-220) and iron seric level 24 microg/dL (60-130). Faecal research highlighted a very scarce presence of E. coli, resulting in 102 UFC/g of stool. Of all enteroinvasive pathogens, researched by molecular analyses, only Yersinia spp. was positive. After several specific cycles of antibiotic and antinflammatory therapies, the patient improved its general health condition considerably and showed almost complete regression of aching inguinal lymph node inflammation. In a picture of a worsening inflammatory process, produced by pathogens like Mycoplasmas, chronic silent or low grade inflammation atypical agents, in young HLA-B27 positive patient, VDRL test resulted positive. This value represents the first non-specific unique spy to reveal the precocious immunological signal in order to register the beginning of early innate immune system decay, keeping in mind that mycoplasmal and chlamydial infections are the triggering of cancer in patients genetically susceptible.

Experimental and modeling study of the reaction C2F4 (+ M) ⇔ CF2 + CF2 (+ M).

The thermal dissociation reaction C2F4(+ M) → 2CF2(+ M) was studied in shock waves monitoring CF2 radicals by their UV absorption. The absorption coefficients as functions of wavelength and temperature were redetermined and are represented in analytical form. Dissociation rate constants as functions of bath gas concentration [M] and temperature, from previous and the present work, are presented analytically employing falloff expressions from unimolecular rate theory. Equilibrium constants are determined between 1200 and 1500 K. The data are shown to be consistent, with a C-C bond energy of 67.5 (±0.5) kcal mol(-1). High-pressure limiting rate constants for dissociation and recombination are found to be unusually small. This phenomenon can be attributed to an unusually pronounced anisotropy of the potential energy surface, such as demonstrated by quantum-chemical calculations of the potential energy surface.

Jugulodigastric lymph node inflammation derived from chronic atypical oropharyngeal phlogosis recurring annually after flu virus vaccination: a holistic vision of a clinical case solved after chlamydicidal antibiotic therapy.

In this report, we evaluated the case history of a patient with longstanding chronic pharyngitis who had periodic clinical manifestation for three years after a flu vaccine administration, and after various treatments tried to resolve the chronic pharyngitis with unsuccessful antibiotic and anti-inflammatory therapies. The patient occasionally presented a slight ocular inflammation, while dysuria occurred after sexual activity. The search for common pathogens by use of pharyngeal swabs resulted only in Corynebacterium ulcerans growth. After this first result, we focused our investigations on ocular and uro-genital infections of Chlamydiaceae (Ct and Cp) and Mycoplasmataceae (Mh and Uu) families. We examined the patient?s pharynx using molecular and culture techniques from three different sites. Although several infectious agents, including viruses and bacteria, causing chronic pharyngitis are reported in the literature, these ocular and uro-genital pathogens are seldomly routinely investigated in the same patient in ORL. Furthermore, while episodes of chronic pharyngitis is one of the most common clinical manifestation in ENT patients, these atypical pharyngitis represent ever-increasing infections which must always be considered and researched by suitable instruments such as PCR. Only from the collection of detailed medical history and careful observations of clinical manifestation, indicative of an oral chronic pathologic phenomenon of low intensity initiated several years previously, starting with sudden outbreak and relapse like a bout of flu, we suggest to study these atypical infecting agents frequently localized in the urogenital human area, awhich would allow to highlight and to recognize these clinical cases that manifest themselves as chronic inflammation of jugulodigastric lymph nodes, remaining still unrecognized and rarely associated to chlamydial infection, confused with the response to flu vaccination. After several specific cycles of antibiotic therapy, the patient's health improved considerably and showed almost complete regression of jugulodigastric lymph node inflammation.

Solitary plasmacytoma of the tonsillar site associated with actinomyces infection: the possible role of IL-6.

ExtraMedullary Plasmacytoma (EMP) is a rare plasma cell tumor. It can occur in the upper aerodigestive tract and presents as a large nodule causing local compressive symptoms. A 79-year old woman presented to Otorhinolaryngology Department with progressive hearing loss and no other symptoms. Following PET/TC examination due to the suspicion of a lymphoproliferative disease, the patient underwent tonsillectomy and the diagnosis of solitary EMP was formulated. In addition to that, the histological examination of the tonsillar tissue revealed large colonies of filamentous bacteria, showing abundant sulphur granules and Splendore-Hoeppli phenomenon; these evidences indicating the presence of a chronic Actinomyces infection. Immunohistochemical analysis demonstrated a marked IL-6 immunoreactivity of the neoplastic plasma cells. Interestingly, a marked IL-6 immunoreactivity was also found in the tissue surrounding the Actinomyces colonies. In the present study we report for the first time a solitary EMP associated with Actinomycosis. It is tempting to speculate that the unsuspected and untreated Actinomyces infection, through chronic IL-6 production, could contribute to the neoplastic transformation of plasma cells.

Malignant peripheral nerve sheath tumour (malignant epithelioid Schwannoma) of the parotid gland.

PROBLEMS/OBJECTIVES: Malignant peripheral nerve sheath tumours are uncommon soft tissue tumours originating from Schwann cells or nerve sheath cells. Malignant epithelioid schwannoma is an aggressive variant of malignant peripheral nerve sheath tumour, and unfortunately is related with a high rate of recurrence and poor prognosis. METHODOLOGY: In this study we present a rare case of malignant epithelioid schwannoma of the parotid gland and we discuss its origin, unusual presentation, and possible treatments options. RESULTS: Patient underwent total parotidectomy without neck dissection. Postoperative radiotherapy and chemotherapy management were not necessary. CONCLUSION: Patients with malignant epithelioid schwannomas typically present with pain and/or rapid enlargement of a pre-existing lesion but the diagnosis of these tumours remains difficult as it is based primarily on clinical suspicion. There is no recognized management for this tumour yet while the prognosis seems to correlate best with the extent of surgical resection (Fig. 4, Ref. 16).

The Eps15 C. elegans homologue EHS-1 is implicated in synaptic vesicle recycling.

Eps15 represents the prototype of a family of evolutionarily conserved proteins that are characterized by the presence of the EH domain, a protein-protein interaction module, and that are involved in many aspects of intracellular vesicular sorting. Although biochemical and functional studies have implicated Eps15 in endocytosis, its function in the endocytic machinery remains unclear. Here we show that the Caenorhabditis elegans gene, zk1248.3 (ehs-1), is the orthologue of Eps15 in nematodes, and that its product, EHS-1, localizes to synaptic-rich regions. ehs-1-impaired worms showed temperature-dependent depletion of synaptic vesicles and uncoordinated movement. These phenotypes could be correlated with a presynaptic defect in neurotransmission. Impairment of EHS-1 function in dyn-1(ky51) worms, which express a mutant form of dynamin and display a temperature-sensitive locomotion defect, resulted in a worsening of the dyn-1 phenotype and uncoordination at the permissive temperature. Thus, ehs-1 and dyn-1 interact genetically. Moreover, mammalian Eps15 and dynamin protein were shown to interact in vivo. Taken together, our results indicate that EHS-1 acts in synaptic vesicle recycling and that its function might be linked to that of dynamin.

Hypocrellin-B acetate as a fluorogenic substrate for enzyme-assisted cell photosensitization.

Photosensitizing molecules (PSs) undergo chemico-physical changes upon addition of suitable substituents, influencing both their photophysical properties and their ability to accumulate into cells. Once inside the cells, the modified PS acts as a fluorogenic substrate: the added substituent is removed by a specific enzyme, restoring the native PS in subcellular sensitive sites. We investigated the photophysical properties and interaction with HeLa cells of Hypocrellin-B (HypB), as native molecule and upon acetate-group addition (HypB-Ac). Chemical modification alters both absorption and fluorescence features of HypB; consequently, the dynamics of the enzyme hydrolysis of HypB-Ac can be monitored through restoring the native HypB spectral properties. At the cellular level, only the HypB emission signal was detected within 5 min of incubation with either HypB or HypB-Ac, allowing a direct comparison of the time courses of their intracellular accumulation. Plateau values were reached within 15 min of incubation with both compounds, the emission signals being significantly higher in HypB-Ac than in HypB treated cells. Consistently, imaging showed a rapid appearance of red fluorescence in the cytoplasm, with more abundant bright spots in HypB-Ac treated cells. Both compounds did not induce dark toxicity at concentrations up to 1 × 10(-6) M, while upon irradiation at 480 nm phototoxicity was significantly higher for cells exposed to HypB-Ac than for HypB-loaded cells. These findings suggest an improved efficacy of acetylated HypB to be internalized by cells through membrane trafficking, with a preferential interaction of the photoactive molecules on sensitive intracellular sites. After irradiation, in HypB-Ac treated cells, prominent disorganization of several cytoplasmic organelles such as the endoplasmic reticulum, Golgi apparatus, lysosomes, microfilaments and microtubules were observed.

Shock wave study of the thermal decomposition of CF3 and CF2 radicals.

The thermal dissociation reactions CF(3) + M --> CF(2) + F + M (reaction 1 ) and CF(2) + M --> CF + F + M (reaction 3 ) were studied behind shock waves. CF(2) radicals were monitored through their UV absorption. By working at very low reactant concentrations, the rate coefficients of the unimolecular processes could be derived. Reaction 1 was investigated between 1600 and 2300 K in the intermediate range of the falloff curves, at approximately 10 times larger bath gas pressures than employed in earlier work (Srinivasan, N. K.; Su, M.-C.; Michael, J. V.; Jasper, A. W.; Klippenstein, S. J.; Harding, L. B. J. Phys. Chem. A 2008, 112, 31). The combination of the two sets of data, together with theoretical modeling, allows one to construct falloff curves and to provide complete representations of the temperature and pressure dependences of the rate coefficients. Reaction 3 was studied in the limiting low-pressure range and, over the range 2900-3800 K, a rate coefficient k(3) = [Ar] 1.6 x 10(15) exp(-48,040 K/T) cm(3) molecule(-1) s(-1) was obtained. Representations of the rate coefficients over the full falloff curves were again derived by theoretical modeling.

Temperature and pressure dependence of the reaction 2CF3 (+ M) <==> C2F6 (+ M).

Limiting low- and high-pressure rate coefficients as well as full falloff curves have been modeled by unimolecular rate theory for the recombination reaction 2CF(3) (+ M) --> C(2)F(6) (+ M) and the reverse dissociation of C(2)F(6). The results are compared with experimental data from the literature. Although there are considerable discrepancies (up to a factor of 5) between various experimental data near 300 K and the database for high temperatures is still limited, we try to conclude on the temperature dependence of the high-pressure rate coefficient. We suggest that there is only a small and probably positive temperature coefficient of the latter quantity. The present theoretical modeling seems to be in agreement with this experimental result, but it is in disagreement with conclusions from earlier theoretical work. The difference is attributed to different empirical assumptions about the anisotropy of the potential. It is shown that nearly all previous experiments (except high-temperature shock wave and very low pressure pyrolysis/photolysis experiments) correspond to nearly limiting high-pressure conditions.

Management of tinnitus: oral treatment with melatonin and sulodexide.

The main problem arising from tinnitus is the disturbance it causes in day-to-day life and disturbance in sleep leading to fatigue and general discomfort. We attempted to study the effect of melatonin in conjunction with Sulodexide as a treatment method for tinnitus and evaluate its effectiveness. We studied 102 patients suffering from tinnitus with a Prospective Randomised Controlled Study conducted in a tertiary care ENT department. After randomisation, 34 patients were treated with melatonin and Sulodexide, another 34 were treated with melatonin alone, and the remaining 34 (control group) were managed without therapy in order to evaluate spontaneous variations in quality of tinnitus. Patients were assessed prospectively with Tinnitus Handicap Inventory and Acufenometry both pre-treatment and post-treatment. Among the patients we studied, we found better results with both Tinnitus Handicap Inventory and Acufenometry in the group who received melatonin and Sulodexide as against melatonin alone. Any improvement was noted in the control group. In conclusion, our opinion is that melatonin in combination with Sulodexide is a viable treatment option for patients suffering from central or sensorineural tinnitus.

Parotid surgery in patients over seventy-five years old.

Malignant parotid tumours are generally rare but become more common in the last decades of life; this increased incidence concerns mainly secondary parotid space involvement from metastasis or direct invasion. During the past five years, we observed and operated upon 84 patients for parotid diseases, performing 86 parotidectomies (2 bilateral operations). The patients included 30 (35.7%) over 75 years old (2 subjects over 90), mean age 80.5 years, object of the present study. In these 30 patients 2 chronic infections were observed, 6 benign tumours and 22 malignant tumours with only 6 primary neoplastic lesions. Overall, 14 extended radical parotidectomies were performed, 13 conservative total parotidectomies, 2 superficial parotidectomies and 1 radical parotidectomy, a few associated with neck dissection and/or radiotherapy. Three patients died in the post-operative period from heart attack. Five patients died from disease (3 from melanomas--2 after 3 years and 1 after 2 years--and 2 from primary carcinomas--1 after 2 years and 1 after 1 year). Six patients died without disease from various causes (2 after 4 years, the others after 3, 2, 1 year and 8 months). Nine patients are alive, 8 NED and 1 with disease in the ethmoid after 2 years, the latter well under control with radiotherapy. This experience implies that surgical treatment of these secondary tumours, which are performed more frequently in old age, exposes the patient to the risk of serious complications (3 patients died in the immediate post-operative period) and stresses the importance of careful evaluation of general conditions. However, surgery seems to have been the treatment indicated also in the patients who subsequently died from the disease, but in whom at least two-year survival was achieved in almost all cases. Unfortunately, because of the advanced age of the patients, the final results are penalized by deaths from natural causes. It is, nevertheless, encouraging that over one third of the patients operated upon are free of disease.

Orbital exenteration in elderly patients: personal experience.

Orbital exenteration is a disfiguring procedure which typically involves removal of the entire contents of the orbit including the periorbita, appendages, eyelids and, sometimes, a varying amount of surrounding skin. This operation is reserved for the treatment of potentially life-threatening malignancies arising from the orbit, paranasal sinuses or periocular skin. The marked increase in the average life span and resulting greater incidence of invasive malignant skin tumours of the face, typical of old age, is the reason for the increased rate of exenterations in elderly patients. The purpose of this report is to describe personal experience regarding 8 operations of orbital exenteration carried out on elderly patients, 6 males and 2 females, age range 66-85 years (mean 75), who came to our observation, from January 2002 to December 2007, on account of cancer (7 cases: 4 basal cell carcinomas; 1 squamous cell carcinoma; 1 fibrosarcoma; 1 melanoma) or infectious inflammatory disease (1 case of rhinocerebral mucormycosis) and were treated with type III orbital exenteration (2 cases) and type IV orbital exenteration (6 cases according to Meyer and Zaoli's classification). The methods used to reconstruct the eye-socket consisted of a full-thickness skin graft in 5 cases, pedicled myocutaneous flaps in 2 cases--a latissimus dorsi muscle flap alone, in one patient, and combined with a pectoralis major muscle flap in another - and a combined lateral-based frontal fasciocutaneous pedicled flap and full-thickness skin graft in the oldest patient. Regarding survival and the local clinical situation, 3 of the 4 patients with basal cell carcinomas are alive and disease-free after 6 years, 2 years and 20 months, respectively, while the oldest patient died of the disease after 10 months. The subject who underwent surgery for squamous cell carcinoma is alive and disease-free after 2 years. The patients with melanoma, fibrosarcoma and mucormycosis died. Although there are various options available for reconstruction, full-thickness skin graft or a pedicled muscolocutaneous flap provide the simplest solution in the elderly population with significant co-morbidities. The final outcome is, in our experience, comparable to that of more complex flap reconstruction, obtaining very good final results with minimal donor site morbility and a reduced operation time.

MicroRNA in Sjögren's Syndrome: Their Potential Roles in Pathogenesis and Diagnosis.

Sjögren's syndrome (SS) or sicca syndrome was described by Swedish ophthalmologist Sjögren in the year 1933 for the first time. The etiology of the SS is multifunctional and includes a combination of genetic predisposition and environmental as well as epigenetic factors. It is an autoimmune disease characterized by features of systemic autoimmunity, dysfunction, and inflammation in the exocrine glands (mainly salivary and lacrimal glands) and lymphocytic infiltration of exocrine glands. In fact, the involvement of lacrimal and salivary glands results in the typical features of dry eye and salivary dysfunction (xerostomia). Only in one-third of the patients also present systemic extraglandular manifestations. T cells were originally considered to play the initiating role in the autoimmune process, while B cells were restricted to autoantibody production. In recent years, it is understood that the roles of B cells are multiple. Moreover, autoantibodies and blood B cell analysis are major contributors to a clinical diagnosis of Sjögren's syndrome. Recently, there has been rising interest in microRNA implication in autoimmunity. Unfortunately, to date, there are only a few studies that have investigated their participation in SS etiopathogenesis. The purpose of this work is to gather the data present in the literature to clarify this complex topic.

The E3 ubiquitin ligase WWP1 sustains the growth of acute myeloid leukaemia.

The E3 ubiquitin ligase (E3) WWP1 is an oncogenic factor implicated in the maintenance of different types of epithelial cancers. The role of WW domain-containing E3 ubiquitin protein ligase 1 (WWP1) in haematological neoplasms remains unknown. Acute myeloid leukaemia (AML) is characterized by the expansion of malignant myeloid cells blocked at different stages of differentiation. Here we report that the expression of WWP1 is significantly augmented in a large cohort of primary AML patients and in AML cell lines, compared with haematopoietic cells from healthy donors. We show that WWP1 inactivation severely impairs the growth of primary AML blasts and cell lines in vitro. In vivo, we observed a reduced leukaemogenic potential of WWP1-depleted AML cells upon transplantation into immunocompromised mice. Mechanistically, WWP1 inactivation induces the accumulation of its protein substrate p27Kip1, which ultimately contributes to G0/G1 cell cycle arrest of AML blasts. In addition, WWP1 depletion triggers the autophagy signalling and reduces survival of leukaemic cells. Collectively, our findings provide molecular insights into the anti-cancer potential of WWP1 inhibition, suggesting that this E3 is a promising biomarker and druggable target in AML.

Can the onset of orbital cancer be the result of a prosthetic eye?

Orbital exenteration is a disfiguring procedure performed for unresponsive orbital infections and control of recurrent benign tumours and malignancies arising from the eyelids (basal cell carcinoma, squamous cell carcinoma, conjunctival malignant melanoma), lachrymal glands (adenoid cystic carcinoma) or surrounding sinuses. In extremely rare cases the use of a prosthetic eye after enucleation can lead to anophthalmic socket tumours. We report the case of a 54-year-old man who had left eye enucleation due to recurring events of retinal detachment and who developed an invasive fast growing epidermoid carcinoma 30 years later. We review the literature to evaluate the rarity of the occurrence, time of onset after enucleation, treatments and outcomes. Our case illustrates the management of the pathology and emphasises the necessity of careful examination of the anophthalmic socket and the ocular prosthesis to identify any irregularities or damage on its surface even after exenteration that is not performed for malignant disease. Long-term follow up is necessary because this tumour could occur at long time periods after enucleation.

Abnormal phenotype of in vitro dermal fibroblasts from patients with Pseudoxanthoma elasticum (PXE).

Pseudoxanthoma elasticum (PXE) is a genetic connective tissue disease, whose gene and pathogenesis are still unknown. Dermal fibroblasts from patients affected by PXE have been compared in vitro with fibroblasts taken from sex and age-matched normal individuals. Cells were grown and investigated in monolayer, into three-dimensional collagen gels and in suspension. Compared with normal cells, PXE fibroblasts cultured in monolayer entered more rapidly within the S phase and exhibited an increased proliferation index; on the contrary, similarly to normal fibroblasts, PXE cells did not grow in suspension. Furthermore, compared with normal fibroblasts, PXE cells exhibited lower efficiency in retracting collagen type I lattices and lower adhesion properties to collagen type I and to plasma fibronectin. This behavior was associated with higher expression of integrin subunits alpha2, alpha5, alphav, whereas beta1 subunit as well as alpha2beta1 and alpha5beta1 integrin expression was lower than in controls. Compared to controls, PXE fibroblasts had higher CAM protein expression in accordance with their high tendency to form cellular aggregates, when kept in suspension. The demonstration that PXE fibroblasts have altered cell-cell and cell-matrix interactions, associated with modified proliferation capabilities, is consistent with the hypothesis that the gene responsible for PXE might have a broad regulatory role on the cellular machinery.

New light in flavin autofluorescence.

Our attention was captured by the interesting debate on the identification of lipofuscins, lipofuscin-like lipopigments, or flavins as the responsible for intracellular autofluorescence (AF) detected in epithelial cancer stem cells when exciting at 480-490 nm. Evidence was provided leading to ascribe AF emission to flavins accumulating in cytoplasmic structures, bounded to membranes and bearing ATP-dependent ABCG2 transporters. Flavins were then proposed as an intrinsic AF biomarker of cancer stem cells, with advantageous implications on cell invasiveness and chemoresistance investigations. It is however worth recalling the huge amount of literature on flavins and NAD(P)H as AF biomarkers of cell energetic metabolism and redox state, an aspect that should not be overlooked in the renewed course to extend the potential of flavins as AF biomarkers, entailing also a recent proposal of Flavin-based fluorescent proteins as substitutes of Green fluorescent proteins.