X-ray irradiation effects on nuclear and membrane regions of single SH-SY5Y human neuroblastoma cells investigated by Raman micro-spectroscopy.

Raman micro-spectroscopy was performed in vitro on nuclear and membrane regions of single SH-SY5Y human neuroblastoma cells after irradiation by graded X-ray doses (2, 4, 6, 8 Gy). The acquired spectra were analyzed by principal component analysis (PCA) and interval-PCA (i-PCA) methods. Biochemical changes occurring in the different regions of single cells as a consequence of the radiation exposure were observed in cells fixed immediately after the irradiation. The most relevant effects arose from the analysis of the spectra from the cell nucleus region. The observed changes were discussed in terms of the modifications in the cell cycle, resulting in an increase in the DNA-related signal, a protein rearrangement and changes in lipid and carbohydrates profiles within the nucleus. Potential markers of an apoptotic process in cell population irradiated with 6 and 8-Gy X-ray doses could have been singled out. No significant effects were found in spectra from cells fixed 24 h after the irradiation, thus suggesting the occurrence of repairing processes of the X-ray induced damage.

Financial impact of surgical training on hospital economics: an income analysis of 1184 out-patient clinic consultations.

INTRODUCTION: In many countries healthcare commissioning bodies (state or insurance-based) reimburse hospitals for their activity. The costs associated with post-graduate clinical training as part of this are poorly understood. This study quantified the financial revenue generated by surgical trainees in the out-patient clinic setting. METHODS: A retrospective analysis of surgical out-patient ambulatory care appointments under 6 full-time equivalent Consultants (Attendings) in one hospital over 2 months. Clinic attendance lists were generated from the Patient Access System. Appointments were categorised as: 'new', 'review' or 'procedure' as per the Department of Health Payment by Results (PbR) Outpatient Tariff (Outpatient Treatment Function Code 104; Outpatient Procedure Code OPRSI1). RESULTS: During the study period 78 clinics offered 1184 appointments; 133 of these were not attended (11.2%). Of those attended 1029 had sufficient detail for analysis (98%). 261 (25.4%) patients were seen by a trainee. Applying PbR reimbursement criteria to these gave a projected annual income of £GBP 218,712 (€EU 266,527; $USD 353,657) generated by 6 surgical trainees (Residents). This is equivalent to approximately £GBP 36,452 (€EU 44,415; $USD 58,943) per trainee annually compared to £GBP 48,732 (€EU 59,378; $USD 78,800) per Consultant. This projected yearly income off-set 95% of the trainee's basic salary. CONCLUSION: Surgical trainees generated a quarter of the out-patient clinic activity related income in this study, with each trainee producing three-quarters of that generated by a Consultant. This offers considerable commercial value to hospitals. Although this must offset productivity differences and overall running costs, training bodies should ensure hospitals offer an appropriate return. In a competitive market hospitals could be invited to compete for trainees, with preference given to those providing excellence in training.

Analysis of occlusal stresses transmitted to the inferior alveolar nerve by multiple threaded implants.

BACKGROUND: Potential nerve injury or loss of sensation can occur after mandibular implant placement or loading. To avoid this type of damage, it is critical to determine the proper distance from implants to the mandibular nerve. Hence, the purpose of this study is to use biomechanical analyses to determine the safe distance from multiple implants to the inferior alveolar nerve. METHODS: Using the boundary element method, a numerical mandibular model was designed to simulate a mandibular segment containing multiple threaded fixtures. This model allows assessment of the pressure, as induced by occlusal loads, on the trigeminal nerve. Such pressure distribution was evaluated against different distances from the fixtures to the mandibular canal, against the possible lack of the central fixture in a three-abutment configuration, and against different levels of implant osseointegration. All the simulations considered a canal that is orthogonal to the implant axis. RESULTS: Nerve pressure increased quickly when the implant-canal distance decreased in the range studied. Lack of the central implant to support the central abutment caused major increases in nerve pressure. CONCLUSIONS: This study suggests a minimal implant-canal distance of 1 mm to prevent inferior alveolar nerve damage caused by three connected implants. For clinical safety, an additional 0.5 mm is recommended as a cushion, so a 1.5-mm minimal distance should be planned to avoid potential nerve injury.

Enzyme distribution and secondary structure of sol-gel immobilized glucose oxidase by micro-attenuated total reflection FT-IR spectroscopy.

Glucose oxidase (GOD) immobilized into sol-gel matrices was studied by using Micro-Attenuated Total Reflection Fourier Transform Infrared (micro-ATR FT-IR) spectroscopy in order to characterize enzyme distribution and secondary structure in systems with valuable potentialities in amperometric and optical biosensing. Spectra were acquired in the 4000-600 cm(-1) frequency region and the analysis of specific fingerprints in the FT-IR spectra evidenced that the enzyme was actually immobilized in the matrix. The enzyme spatial distribution was obtained by examining the amide I and amide II band region of spectra from defined sample positions. The deconvolution of the amide I band in terms of lorentzian functions provided information on the secondary structure of the immobilized GOD. By this approach a macroscopic preservation of GOD activity upon immobilization was evidenced along with the existence of some matrix sites with locally inactivated GOD. To our knowledge this is the first example of point-by-point characterization of conformational changes of immobilized enzyme by means of micro-ATR infrared spectroscopy, thus confirming that this technique can be usefully employed for a non- or minimally-invasive detailed micro-characterization of catalytic supports in order to improve their functionality.

Has selenium a chemopreventive effect on hepatocellular carcinoma?

Hepatocellular carcinoma (HCC) is one of the most lethal cancers in the world. Its etiology includes chronic liver disease, viral hepatitis, alcoholism, and hepatic cirrhosis. Both oxidative stress and inflammatory mechanisms have been implicated in HCC pathophysiology. Surgical resection and liver transplants are currently used to treat HCC. Consequently, there exists a decisive requirement to explore possible alternative chemopreventive and therapeutic strategies for HCC. The use of dietary antioxidants and micronutrients has been proposed as a useful means for the HCC management. Trace elements such as selenium are involved in several major metabolic pathways as well as antioxidant defense systems. In particular, selenium is an important oligo-element that plays a central role in cellular redox processes even if the amount necessary for the cell functions is in a very narrow range. However, selenium is involved in the prevention of numerous chronic diseases and cancers. This review will examine the potential role of selenium in HCC prevention and treatment and, in detail, focus on: i) description of selenium in biological systems and in mammalian proteins, ii) involvement of selenium in HCC, iii) in vivo and in vitro effects of selenium in preclinical models of HCC and iv) potential challenges involved in the selenium use in the prevention and treatment of HCC.

An acetylcholinesterase biosensor for determination of low concentrations of Paraoxon and Dichlorvos.

The characterization of an economic and ease-to-use carbon paste acetylcholinesterase (AChE) based biosensor to determine the concentration of pesticides Paraoxon and Dichlorvos is discussed. AChE hydrolyses acetylthiocholine (ATCh) in thiocoline (TC) and acetic acid (AA). When AChE is immobilized into a paste carbon working electrode kept at +410 mV vs. Ag/AgCl electrode, the enzyme reaction rate using acetylthiocholine chloride (ATCl) as substrate is monitored as a current intensity. Because Paraoxon and Dichlorvos inhibit the AChE reaction, the decrease of the current intensity, at fixed ATCl concentration, is a measure of their concentration. Linear calibration curves for Paraoxon and Dichlorvos determination have been obtained. The detection limits resulted to be 0.86 ppb and 4.2 ppb for Paraoxon and Dichlorvos, respectively, while the extension of the linear range was up 23 ppb for the former pesticide and up to 33 ppb for the latter. Because the inhibited enzyme can be reactivated when immediately treated with an oxime, the biosensor reactivation has been studied when 1,1'-trimethylene bis 4-formylpyridinium bromide dioxime (TMB-4) and pyridine 2-aldoxime methiodide (2-PAM) were used. TMB-4 resulted more effective. The comparison with the behavior of similar AChE based biosensors is also presented.

Bisphenol A content in fish caught in two different sites of the Tyrrhenian Sea (Italy).

Bisphenol A (BPA) is an endocrine disruptor (ED) that is abundant in the environment because of its extensive use in human-manufactured products. In this study, the BPA concentration was measured in the muscle and liver of five edible fish, characterized by different habitat and habits, caught in two different sites of the Tyrrhenian Sea (Italy). Our results show that: (i) fish livers are about 2.5 times more polluted than muscle; (ii) fish caught in the Gulf of Naples are more polluted than those from the Latium coasts, ranging from 1.2-fold more for White Bream to 6.6-fold for Grey Mullet; and (iii) the percentages of fish found to be BPA-polluted in the Gulf of Naples ranged from 73% (for Bass) to 90% (for Mullet), while the Latium fish range from 60% (for Bass) to 90% (for Mullet). These data indicate that consumers of fish caught in the Gulf of Naples are at a greater risk for BPA-induced endocrine pathologies compared to those who consume fish caught along the Latium coasts.

Visible micro-Raman spectroscopy for determining glucose content in beverage industry.

The potential of Raman spectroscopy with excitation in the visible as a tool for quantitative determination of single components in food industry products was investigated by focusing the attention on glucose content in commercial sport drinks. At this aim, micro-Raman spectra in the 600-1600cm(-1) wavenumber shift region of four sport drinks were recorded, showing well defined and separated vibrational fingerprints of the various contained sugars (glucose, fructose and sucrose). By profiting of the spectral separation of some peculiar peaks, glucose content was quantified by using a multivariate statistical analysis based on the interval Partial Least Square (iPLS) approach. The iPLS model needed for data analysis procedure was built by using glucose aqueous solutions at known sugar concentrations as calibration data. This model was then applied to sport drink spectra and gave predicted glucose concentrations in good agreement with the values obtained by using a biochemical assay. These results represent a significant step towards the development of a fast and simple method for the on-line glucose quantification in products of food and beverage industry.

A thionine-modified carbon paste amperometric biosensor for catechol and bisphenol A determination.

A thionine-modified carbon paste electrode for catechol and Bisphenol A (BPA) detection is presented. Graphite powder was modified by adsorbing thionine as electrochemical mediator. The electrochemical response of the modified carbon paste electrode (CPE) was determined before electrode modification with tyrosinase. Then, tyrosinase was added in order to assemble a biosensor. Once established the best operative conditions, an interelectrode reproducibility around 7% was obtained and the resulting biosensor showed improved sensitivities and (S=139.6+/-1.1 nA/microM for catechol and S=85.4+/-1.5 nA/microM for BPA) in comparison with the biosensor constructed without thionine (S=104.4+/-0.5 nA/microM for catechol and S=51.1+/-0.6 nA/microM for BPA) and low detection limits (0.15 microM for both the electrodes and analytes). Also the comparison with the results reported in the literature showed higher sensitivity and lower detection limit for our biosensor. Moreover the functioning of the thionine-tyrosinase CPE was validated following a biodegradation process of water polluted by BPA and comparing the time changes of BPA concentration inferred by the biosensor calibration curve and those determined by means of HPLC measurements.

Forensic sciences and forensic odontology: issues for dental hygienists and therapists.

The scientific literature contains very little about the role of the dental hygienist/therapist in the specific areas of forensic investigations and collection of evidence. The authors examine how the contribution of a highly qualified dental hygienist can be particularly helpful during human forensic identification operations and non-accidental traumas like domestic violence, child abuse, neglect and bitemarks. Forensic dental identification of human remains is a highly complex multidisciplinary challenge. It requires the involvement of several professionals who are expert in forensic science. Among these, one or more adequately trained dental hygienists could be involved. Dental hygienists/therapists may also be asked to record cutaneous lesions in two different situations. The first may be the dental office where she/he may detect oval, elliptic, or semicircular lesions on the skin of the uncovered neck, shoulder and arms of a patient. The second is the crime scene or the morgue (if one is involved), which may require a visit by the forensic odontologist called by the medical examiner or the coroner to perform an odontological autopsy. The purpose of our study is to highlight procedures that should be followed by the dental hygienist/therapist in collecting forensic information in the above-mentioned scenarios. As a valuable resource, the authors recommend training of dental hygienists in the area of forensic sciences, with particular attention to information technology and photography.

Liver X receptor activation controls intracellular cholesterol trafficking and esterification in human macrophages.

Liver X receptors (LXRs) are nuclear receptors that regulate macrophage cholesterol efflux by inducing ATP-binding cassette transporter A1 (ABCA1) and ABCG1/ABCG4 gene expression. The Niemann-Pick C (NPC) proteins NPC1 and NPC2 are located in the late endosome, where they control cholesterol trafficking to the plasma membrane. The mobilization of cholesterol from intracellular pools to the plasma membrane is a determinant governing its availability for efflux to extracellular acceptors. Here we investigated the influence of LXR activation on intracellular cholesterol trafficking in primary human macrophages. Synthetic LXR activators increase the amount of free cholesterol in the plasma membrane by inducing NPC1 and NPC2 gene expression. Moreover, ABCA1-dependent cholesterol efflux induced by LXR activators was drastically decreased in the presence of progesterone, which blocks postlysosomal cholesterol trafficking, and reduced when NPC1 and NPC2 mRNA expression was depleted using small interfering RNA. The stimulation of cholesterol mobilization to the plasma membrane by LXRs led to a decrease in cholesteryl ester formation and Acyl-coenzyme A cholesterol acyltransferase-1 activity. These data indicate that LXR activation enhances cholesterol trafficking to the plasma membrane, where it becomes available for efflux, at the expense of esterification, thus contributing to the overall effects of LXR agonists in the control of macrophage cholesterol homeostasis.

Peroxisome proliferator-activated receptor alpha controls cellular cholesterol trafficking in macrophages.

The mobilization of cholesterol from intracellular pools to the plasma membrane is a determinant that governs its availability for efflux to extracellular acceptors. NPC1 and NPC2 are proteins localized in the late endosome and control cholesterol transport from the lysosome to the plasma membrane. Here, we report that NPC1 and NPC2 gene expression is induced by oxidized LDL (OxLDL) in human macrophages. Because OxLDLs contain natural activators of peroxisome proliferator-activated receptor alpha (PPARalpha), a fatty acid-activated nuclear receptor, the regulation of NPC1 and NPC2 by PPARalpha and the consequences on cholesterol trafficking were further studied. NPC1 and NPC2 expression is induced by synthetic PPARalpha ligands in human macrophages. Furthermore, PPARalpha activation leads to an enrichment of cholesterol in the plasma membrane. By contrast, incubation with progesterone, which blocks postlysosomal cholesterol trafficking, as well as NPC1 and NPC2 mRNA depletion using small interfering RNA, abolished ABCA1-dependent cholesterol efflux induced by PPARalpha activators. These observations identify a novel regulatory role for PPARalpha in the control of cholesterol availability for efflux that, associated with its ability to inhibit cholesterol esterification and to stimulate ABCA1 and scavenger receptor class B type I expression, may contribute to the stimulation of reverse cholesterol transport.

Modulation of the catalytic activity of free and immobilized peroxidase by extremely low frequency electromagnetic fields: dependence on frequency.

A study of the influence of electromagnetic fields (EMF) of various frequencies, from 50 up to 400 Hz, on the catalytic activity of soluble and insoluble horseradish peroxidase (POD) was carried out. To simulate the conditions in which the enzyme operates in vivo, the POD was immobilized by entrapment on a gelatin membrane or by covalent attachment on a nylon graft membrane. The rate of inactivation of the soluble POD was found to exhibit positive and negative interactions with the 1 mT applied magnetic field, with an optimum positive effect at 130 Hz. The immobilized PODs, on the contrary, do not exhibit negative interactions, but show a maximum positive interaction at 150 Hz when entrapped and at 170 Hz when covalently attached. At 50 Hz and at frequencies higher than 250 Hz no effects were observed with insoluble POD. The optimum frequency of positive interaction between the EMF and the catalytic activity of the insoluble enzymes is shifted with respect to that of the soluble enzymes towards higher frequencies, the size of the shifts being dependent on the intensity of the physical forces involved in the immobilization process.

Er:YAG laser treatments on dentine surface: micro-Raman spectroscopy and SEM analysis.

OBJECTIVE: This study aims to give an insight into the chemical and structural modifications occurring in laser treated dentine surface, as a tool to plan a new class of dental materials specific for the dental restoration when laser technologies are used. For this purpose the effect of Er:YAG (Erbium-Yttrium Aluminium Garnet) laser treatments on human dentine surface has been investigated. METHODS: The dentine surface of periodontally compromised teeth cut along the main axis was treated in vitro with laser pulses having the same energy but time duration varying in the range of 100-1000 micros. The chemical characteristics and the morphology of the laser treated surface have been studied with micro-Raman spectroscopy (micro-RS) and scansion electron microscopy (SEM). RESULTS: The micro-RS and SEM analysis of laser treated teeth have been compared with those of surfaces treated with conventional mechanical methodologies. The laser pulse time duration affects the surface morphology and the chemistry of dentine, mainly of organic components. The effects have been monitored by analysing the C-H group stretch vibration mode of the micro-RS spectrum. CONCLUSION: The laser treatment can determine different chemical microenvironments on the treated dentine surface, depending on the time duration of laser pulse used. Treatments with very long pulse results in a the dentine surface very similar to that obtained with a conventional mechanical drill, while with very short laser pulses a strong modification of the dentine organic components has been observed.

Time-resolved contrast function and optical characterization of spatially varying absorptive inclusions at different depths in diffusing media.

The role of a spatially varying absorptive inhomogeneity located at different depths within a turbid material has been investigated. This inhomogeneity has been characterized by a spatially dependent Gaussian distribution of its absorption coefficient. The present study has been performed calculating the time-resolved contrast function in the framework of the first-order perturbative approach to the diffusion equation for a slab geometry and a coaxial measurement scheme. The model has allowed us to take into account different locations of the inclusion along the source-detector axis. The accuracy of time-resolved contrast predictions has been analyzed through comparisons with results of the finite element method that has been used to numerically solve the diffusion equation. Recovery of the absorption perturbation parameter of the inhomogeneity for different axial positions has also been investigated.

Perturbation model to predict the effect of spatially varying absorptive inhomogeneities in diffusing media.

We develop a perturbation model to predict the effect of a spatially varying absorptive inhomogeneities in a diffusing slab. The model is based on a perturbation solution of diffusion equation derived for a refractive index mismatch between the scattering slab and the surrounding medium, through the use of the extrapolated boundary conditions. We show that the model allows to compute the time-dependent relative change in the transmitted signal resulting from the presence of the inclusion. We derive simplified expressions for the perturbed time-resolved transmittance that allows to implement an efficient fitting procedure for obtaining the optical properties of the absorptive inclusion. The accuracy of the predictions of the model was investigated through comparison with the results of the Finite Element Method to solve the time-dependent diffusion equation numerically. The procedure is used to obtain the absorption perturbation parameter of an absorptive inclusion characterized by spatially dependent Gaussian distribution of its absorption coefficient located at the midplane of a scattering slab.

In vitro studies of the influence of ELF electromagnetic fields on the activity of soluble and insoluble peroxidase.

The influence of an extremely low frequency (ELF) magnetic field (50 Hz and 1 mT, EMF) on the activity of a soluble and insoluble horseradish peroxidase (E.C. 1.11.17) has been studied as a function of time. Insoluble derivatives were obtained by enzyme entrapment into two different gelatin membranes or by covalent attachment of the enzyme on two nylon membranes, differently preactivated. Results have shown that the field affects the inactivation rate of the soluble enzyme, while no effects are observed with insoluble derivatives. Since in vivo enzymes are immobilised into the biomembrane bilayer or entrapped into the cytoplasmic mixture, one might speculate that our experimental conditions do not reflect the catalytic activity of the enzymes in vivo.

Depth dependence of the analytical expression for the width of the point spread function (spatial resolution) in time-resolved transillumination.

Simple analytical expressions for the point spread function (PSF) at different depths can save computation time and improve the performance of inverse algorithms for optical imaging. In particular, application of such formulas simplifies quantification of the optical characteristics of tissue abnormalities inside highly scattering media. Earlier it was shown within the random walk theory framework that the PSF for time-resolved transillumination imaging through a highly scattering slab is well represented by a Gaussian. We have experimentally validated a simple formula of the random walk model for the effective width of this Gaussian, as a function of time delay, at different depths of the target. Presented analysis of published experimental data, concerning effective width of the PSF, for a slab of considerably smaller thickness also demonstrates good agreement between the data and predictions of our model. This PSF width determines spatial resolution of the time-resolved imaging and is widely discussed in the literature.

On selective left neglect during walking in a child.

In this article we describe a child affected by right parieto-occipital lesion due to head injury. The patient showed left hemianopia, but not unilateral spatial neglect on traditional paper-and-pencil tests and on "ecological" tests. However, his parents reported frequent collisions with obstacles on the left side. A specific test was set up: The patient had to kick down skittles put on both sides of a route traced on the floor. He kicked down 89% of skittles on the right, but only 38% on the left side. These findings are discussed in light of recent theories on unilateral neglect.