Thioalbamide inhibits FoF1-ATPase in breast cancer cells and reduces tumor proliferation and invasiveness in breast cancer in vivo models.

OBJECTIVE: Thioalbamide is a ribosomally synthesized and post-translationally modified peptide (RiPP) belonging to the family of thioamitides, a rare class of microbial specialized metabolites with unusual post-translational modifications and promising biological activities. Recent studies have demonstrated the ability of thioalbamide to exert highly selective cytotoxic effects on tumor cells by affecting their energy metabolism, thus causing abnormal ROS production and triggering apoptosis. This study is aimed to investigate the molecular mechanisms underlying the antitumor activity of thioalbamide in order to identify its exact molecular target. METHODS: Wild type MCF-7 and MDA-MB-231 breast cancer cell lines as well as cancer cells deprived of mitochondrial DNA (ρ0 cells) were employed in order to assess thioalbamide effects on tumor bioenergetics. In this regard, metabolic profile was evaluated by a Seahorse XFe96 analyzer, and the activity of the enzyme complexes involved in oxidative phosphorylation was quantified by spectrophotometric assays. Thioalbamide effects on tumor invasiveness were assessed by gelatin zymography experiments and invasion assays. In vivo experiments were carried out on breast cancer xenograft and "experimental metastasis" mouse models. RESULTS: Experiments carried out on ρ0 breast cancer cells, together with Seahorse analysis and the application of spectrophotometric enzymatic assays, highlighted the ability of thioalbamide to affect the mitochondrial respiration process, and allowed to propose the FoF1-ATPase complex as its main molecular target in breast cancer cells. Additionally, thioalbamide-mediated OXPHOS inhibition was shown, for the first time, to reduce tumor invasiveness by inhibiting metalloproteinase-9 secretion. Furthermore, this study has confirmed the antitumor potential of thioalbamide in two different in vivo models. In particular, experiments on MCF-7 and MDA-MB-231 xenograft mouse models have confirmed in vivo its high anti-proliferative and pro-apoptotic activity, while experiments on MDA-MB-231 ″experimental metastasis" mouse models have highlighted its ability to inhibit breast cancer cell invasiveness. CONCLUSIONS: Overall, our results shed more light on the molecular mechanisms underlying the pharmacological potential of thioamidated peptides, thus reducing the gap that separates this rare class of microbial metabolites from clinical studies, which could validate them as effective tools for cancer treatment.

Involvement of transcribed lncRNA uc.291 and SWI/SNF complex in cutaneous squamous cell carcinoma.

While non-melanoma skin cancers (NMSCs) are the most common tumours in humans, only the sub-type cutaneous squamous cell carcinoma (cSCC), might become metastatic with high lethality. We have recently identified a regulatory pathway involving the lncRNA transcript uc.291 in controlling the expression of epidermal differentiation complex genes via the interaction with ACTL6A, a component of the chromatin remodelling complex SWI/SNF. Since transcribed ultra-conserved regions (T-UCRs) are expressed in normal tissues and are deregulated in tumorigenesis, here we hypothesize a potential role for dysregulation of this axis in cSCC, accounting for the de-differentiation process observed in aggressive poorly differentiated cutaneous carcinomas. We therefore analysed their expression patterns in human tumour biopsies at mRNA and protein levels. The results suggest that by altering chromatin accessibility of the epidermal differentiation complex genes, down-regulation of uc.291 and BRG1 expression contribute to the de-differentiation process seen in keratinocyte malignancy. This provides future direction for the identification of clinical biomarkers in cutaneous SCC. Analysis of publicly available data sets indicates that the above may also be a general feature for SCCs of different origins.

An ancient remedial repurposing: synthesis of new pinocembrin fatty acid acyl derivatives as potential antimicrobial/anti-inflammatory agents.

Five new pinocembrin derivatives (MC1-MC5) were synthesized by Steglich reaction, and investigated for their antimicrobial, antioxidant, and anti-inflammatory activity. MC2 (oleoyl derivative) and MC3 (linoleoyl derivative) have shown the highest inhibitory effects on bacterial proliferation, with MIC values of 32 µg/mL against Staphylococcus aureus. The docosahexaenoyl derivative MC5 displayed the highest anti-inflammatory activity, decreasing NO production in LPS-stimulated macrophages with an IC50 value of 15.51 µg/mL higher than the positive control diclofenac (IC50 of 39.71 µg/mL). All new synthesized compounds showed no anti-proliferative effects on RAW 264.7 cells. Results demonstrated as the introduction of fatty acid substituents improved the biological profile of pinocembrin. Moreover, the chemical nature of substituents significantly affects the bioactivity. These preliminary results outline the importance to investigate the synthesis of pinocembrin fatty acids derivatives as new and safe anti-microbial/anti-inflammatory agents.

Corrigendum to: Phosphonium Salt Displays Cytotoxic Effects Against Human Cancer Cell Lines.

Due to an oversight one of the author's name was published wrong in the article entitled "Phosphonium Salt Displays Cytotoxic Effects Against Human Cancer Cell Lines" in "Anti-Cancer Agents in Medicinal Chemistry, 2015, Vol. 17, No. 13. pp. 1796."The correct names of all authors are given below:Dhanyalayam D, Palma G, Cappello AR, Mariconda A, Sinicropi MS, Giordano F, Del Vecchio V, Ramunno A, Arra C, Longo P, Saturnino C.

Phosphonium Salt Displays Cytotoxic Effects Against Human Cancer Cell Lines.

Aims/Objective: Phosphonium salts are compounds whose structural characteristics enable them to cross the plasma and mitochondrial membrane with ease. Cancer cells have higher plasma membrane potentials than normal cells; phosphonium salts selectively accumulate in the mitochondria of neoplastic cells and inhibit mitochondrial function. METHOD: In the present work, we investigated the cytotoxic activity of lipophilic phosphonium salt (11- methoxy11-oxo-undecyl) triphenylphosphonium bromide (MUTP) as well as of the two new phosphine oxide salts, 3,3'-(methylphosphoryl) dibenzenaminium chloride (SBAMPO) and 3,3' (phenylphosphoryl) dibenzenaminium chloride (SBAPPO) on the proliferation of breast cancer cell line (MCF-7) and human uterin cervix adenocarcinoma cells (HeLa). RESULT: We showed that only MUTP exhibits antiproliferative effects on both cell lines, without affecting the normal breast epithelial cell proliferation. More specifically, we demonstrated that MUTP treatment of breast cancer cells is associated with impaired cell-cycle progression and metabolically induces mitochondrial damage and triggers apoptotic cell death in MCF-7 and HeLa cells. Taken together, these findings suggest that MUTP may be capable of selectively targeting neoplastic cell growth and therefore has potential applications as anticancer agent.

Bergamot (Citrus bergamia Risso) Flavonoids and Their Potential Benefits in Human Hyperlipidemia and Atherosclerosis: an Overview.

Elevated serum cholesterol, triglycerides and LDL levels are often associated with an increased incidence of atherosclerosis and coronary artery disease. The most effective therapeutic strategy against these diseases is based on statins administration, nevertheless some patients, especially those with metabolic syndrome fail to achieve their recommended LDL targets with statin therapy, moreover, it may induce many serious side effects. Several scientific studies have highlighted a strong correlation between diets rich in flavonoids and cardiovascular risk reduction. In particular, Citrus bergamia Risso, also known as bergamot, has shown a significant degree of hypocholesterolemic and antioxidant/radical scavenging activities. In addition, this fruit has attracted considerable attention due to its peculiar flavonoid composition, since it contains some flavanones that can act as natural statins. Hence, the study of bergamot flavonoids as metabolic regulators offers a great opportunity for screening and discovery of new therapeutic agents. Cholesterol metabolism, flavonoid composition and potential therapeutic use of C. bergamia Risso will be discussed in the following review.

[Retrospective study on the effects of a whey protein concentrate on body composition in 262 sarcopenic tube fed patients]. / Studio retrospettivo sugli effetti di un concentrato di proteine del siero del latte sulla composizione corporea di 262 pazienti in nutrizione enterale affetti da sarcopenia.

AIM: As pointed in ESPEN guidelines on the use of bioelectrical impedance analysis (BIA), "body cell mass (BCM) is the protein rich compartment which is affected in catabolic states, and loss of BCM is associated with poor clinical outcome". Whey proteins are known to improve lean mass in many conditions. We retrospectively evaluated the effects of a WP concentrate with high cysteine content (WPHCC) on BCM of 262 sarcopenic tube fed patients (pts). METHODS: Two hundred sixty-two sarcopenic tube fed pts (130 males, mean age 68,1±15,6 years) were given daily supplemental WPHCC (0.7±0.2 g/kg body weight) after their usual feeding formula, with the aim to improve their BCM. Each patient received a multifrequency impedance test before and after the beginning of WPHCC supplementation (mean follow-up: 4.2±2.8 months). Fifty percent of patients were affected by neurodegenerative diseases, 36% by cancer, 14% by other conditions. RESULTS: BCM, body weight and fat mass significantly improved (P<0.01) after treatment. No severe side effects were recorded. A slight increase in blood urea was observed. CONCLUSION: In our population WPHCC have been safe and effective in improving BCM. WPHCC could be useful to improve BCM in sarcopenic tube fed pts, although renal function should be monitored.

Strength of zirconia fixed partial dentures: review of the literature.

OBJECTIVE: The aim of this study is a systematic review of the literature on the strength and long-term behavior of zirconia FPDs. METHODS: The literature search was performed using as the primary source the Medline database. Were also imposed a set of inclusion and exclusion criteria in order to narrow the search to differentiate the two distinct parts in which it was structured the review. In addition, MeSH terms were applied to further refine the choice of articles more relevant to the review. In the part of the review concerning the long-term behavior of zirconia fixed partial dentures were considered useful only those studies with a minimum follow-up of one year. RESULTS: The search provided a total of 813 articles, but only 25 were considered for the review because they were the only ones who satisfied search criteria. In particular, 13 items relate to the mechanical strength of zirconia prostheses and 12 regarding the long term behavior of zirconia FPDs. The resistance values also exceed the 2000 N while the survival rate reaches to almost 94%. CONCLUSIONS: Based on this review, Zirconia FPDs can be considered reliable products in both the anterior and posterior fields with survival rates comparable to traditional metal-ceramic prostheses.

Energetic assessment of trunk postural modifications induced by a wearable audio-biofeedback system.

This paper investigates the trunk postural modifications induced by a wearable device which assesses the trunk sway and provides biofeedback information through sonification of trunk kinematics. The device is based on an inertial wearable sensing unit including three mono-axial accelerometers and three rate gyroscopes embedded and mounted orthogonally. The biofeedback device was tested on nine healthy subjects during quiet stance in different conditions of sensory limitation eyes closed on solid surface, eyes open on foam cushion surface, eyes closed on foam cushion surface. Five trials were performed for each condition; the order of the trials was randomized. The results reported in this paper show how subjects reduced their rotational kinetic energy by using the biofeedback information and how this reduction was related to the limitation of sensory information.

Light induced atomic desorption from dry-film coatings.

We report the first experimental evidence of nonthermal light induced atomic desorption (LIAD) from octadecyltrichlorosilane dry film. The experiment has been made with Rb confined in a coated cell kept at room temperature. A detailed study of the main features of LIAD effect has been made by varying intensity and wavelength of desorbing light. A discussion about the differences and similarities with other organic films that were studied first is reported. This result is important as it expands the list of materials showing such an effect and increases the possibilities to get suitable light controlled atomic sources for spectroscopy and applications. In particular, we plan to exploit this feature in a Fr magneto-optical trap apparatus.

MicroCT examination of human bone specimens: effects of polymethylmethacrylate embedding on structural parameters.

X-ray microtomography permits the nondestructive investigation of trabecular and cortical bone specimens without special preparation of the sample. To do a quantitative characterization, the cross-section images have to be binarized, separating bone from nonbone. For this purpose, a widely used method is uniform thresholding. However, for commonly available microtomography scanners which use a polychromatic X-ray source, it is unclear what effect the surrounding medium (e.g. air, saline solution, polymethylmethacrylate) has on the threshold value used for the binarization. In the literature an easy procedure to find the optimal uniform threshold value for a given acquisition condition is reported. By applying this procedure, the present work investigated whether a microtomography scan of trabecular bone samples in air or embedded in polymethylmethacrylate gave the same results in terms of structural parameters. The gold standard, that is, histological sections, was used as a reference. Two fixed threshold values were found, one for the microtomography scans performed in air and one for the scans with the same samples embedded in polymethylmethacrylate. These were applied on the correspondent microtomography images for the estimation of structural parameters, such as bone volume fraction, direct trabecular thickness, direct trabecular separation and structure model index. Paired comparisons were made in bone volume fraction between histological sections and microtomography cross-sections for the same bone samples scanned first in air and then embedded in polymethylmethacrylate, by which no significant differences were found. Paired comparisons were also made in bone volume fraction, direct trabecular thickness, direct trabecular separation and structure model index for the same samples over volumes of interest of 4 x 4 x 4 mm3 between microtomography scans in air and scans with the samples embedded in polymethylmethacrylate. Neither these comparisons showed significant differences. This leads to the conclusion that structural parameters estimated by microtomography for human trabecular bone samples scanned either in air or embedded in polymethylmethacrylate are not affected by the surrounding medium (i.e. presence or absence of polymethylmethacrylate), provided that the corresponding optimal threshold value is applied for each acquisition condition.

Human postural response to lower leg muscle vibration of different duration.

Body lean response to bilateral vibrations of soleus muscles were investigated in order to understand the influence of proprioceptive input from lower leg in human stance control. Proprioceptive stimulation was applied to 17 healthy subjects by two vibrators placed on the soleus muscles. Frequency and amplitude of vibration were 60 Hz and 1 mm, respectively. Vibration was applied after a 30 s of baseline. The vibration duration of 10, 20, 30 s respectively was used with following 30 s rest. Subjects stood on the force platform with eyes closed. Postural responses were characterized by center of pressure (CoP) displacements in the anterior-posterior (AP) direction. The CoP-AP shifts as well as their amplitudes and velocities were analyzed before, during and after vibration. Vibration of soleus muscles gradually increased backward body tilts. There was a clear dependence of the magnitude of final CoP shift on the duration of vibration. The amplitude and velocity of body sway increased during vibration and amplitude was significantly modulated by duration of vibration as well. Comparison of amplitude and velocity of body sway before and after vibration showed significant post-effects. Presented findings showed that somatosensory stimulation has a long-term, direction-specific influence on the control of postural orientation during stance. Further, the proprioceptive input altered by soleus muscles vibration showed significant changes in postural equilibrium during period of vibration with interesting post-effects also.

A physical phantom for the calibration of three-dimensional X-ray microtomography examination.

X-ray microtomography is rapidly gaining importance as a non-destructive investigation technique, especially in the three-dimensional examination of trabecular bone. Appropriate quantitative three-dimensional parameters describing the investigated structure were introduced, such as the model-independent thickness and the structure model index. The first parameter calculates a volume-based thickness of the structure in three dimensions independent of an assumed structure type. The second parameter estimates the characteristic form of which the structure is composed, i.e. whether it is more plate-like, rod-like or even sphere-like. These parameters are now experiencing a great diffusion and are rapidly growing in importance. To measure the accuracy of these three-dimensional parameters, a physical three-dimensional phantom containing different known geometries and thicknesses, resembling those of the examined structures, is needed. Unfortunately, such particular phantoms are not commonly available and neither does a consolidated standard exist. This work describes the realization of a calibration phantom for three-dimensional X-ray microtomography examination and reports an application example using an X-ray microtomography system. The calibration phantom (external size 13 mm diameter, 23 mm height) was based on various aluminium inserts embedded in a cylinder of polymethylmethacrylate. The inserts had known geometries (wires, foils, meshes and spheres) and thicknesses (ranging from 20 microm to 1 mm). The phantom was successfully applied to an X-ray microtomography device, providing imaging of the inserted structures and calculation of three-dimensional parameters such as the model-independent thickness and the structure model index. With the indications given in the present work it is possible to design a similar phantom in a histology laboratory and to adapt it to the requested applications.

Device to measure intra-operatively the primary stability of cementless hip stems.

The primary stability of cementless prostheses is critical for the long-term outcome of the operation. Surgeons are currently driven only by their experience in evaluating the extent of stem stability achieved. The aim of the present work was to develop a new device that enables the stability of a cementless stem to be quantitatively assessed intraoperatively. The angle of the stem/femur rotation under torsion and the torque are acquired and compared in real-time to a pre-set threshold inferred from the literature. The device indicates whether the stem is stable or not. It was extensively tested and finally validated in vitro on cadaveric and composite femurs hosting different sizes of the same kind of prostheses, implanted with different levels of press-fitting. The overall accuracy (23%) takes into account not only the overall measurement error but also the variability due to differences in bone quality and stem press-fitting. This error was deemed sufficient to discriminate between stable and unstable implants.

Numerical model to predict the long-term mechanical stability of cementless orthopaedic implants.

The objective of this research was to develop a purely biomechanical model, intended to predict the long-term secondary stability of the implant starting from the biomechanical stability immediately after the operation. A continuous rule-based adaptation scheme was formulated as a dynamic system, and the work verified if such a model produced unique and clinically meaningful solutions. It also investigated whether this continuous model provided results comparable with those of a simpler, discrete-states model used in a previous study. The proposed model showed stable convergence behaviour with all investigated initial conditions, with oscillatory behaviour limited to the first steps of the simulation. The results obtained with the wide range of initial conditions support the hypothesis of the existence and uniqueness of the solution for all initial conditions. The differences between the continuous model and the simpler and more efficient finite-states model were found to be extremely modest (less than 4% over the predicted bonded area). Because of these minimal differences, the use of the much faster finite-states model is recommended to investigate asymptotic conditions, and the continuous model described should be used to investigate the evolution over time of the adaptive process.

Periodical in-situ re-calibration of force platforms: a new method for the robust estimation of the calibration matrix.

The paper provides a new technique based on a least-squares approach for the accurate estimation of a force platform calibration matrix using simple manual procedures, when the direction of the applied loads cannot be perfectly aligned with the axes of the platform. This new procedure can be applied to all force platforms and allows the combined application of vertical and horizontal forces, both static and time-varying. The robust calibration method includes the angular errors in the least-squares parameter vector, thus reducing the bias in the estimated calibration matrix parameters. The performance of the robust method was compared with the conventional one, using a numerical simulation approach starting from a known calibration matrix. With the conventional approach, in noiseless conditions, the maximum error due to load misalignment (SD = 3 degrees) was 6% for the direct terms and over 10% for the cross-talk terms. With the robust method, these errors reduced to zero and were always below 0.4%, even when realistic noise was superimposed on the measures. With perfectly aligned loads and realistic output noise, the confidence intervals of the calibration matrix parameters were very similar for the two methods, demonstrating that the increased number of parameters did not affect the reliability of the estimate.

Feature selection of stabilometric parameters based on principal component analysis.

This study addresses the challenge of identifying the features of the Centre of pressure (COP) trajectory that are most sensitive to postural performance, with the aim of avoiding redundancy and allowing a straightforward interpretation of the results. Postural sway in 50 young, healthy subjects was measured by a force platform. Thirty-seven stabilometric parameters were computed from the one-dimensional and two-dimensional COP time series. After normalisation to the relevant biomechanical factors, by means of multiple regression models, a feature selection process was performed based on principal component analysis. Results suggest that COP two-dimensional time series can be primarily characterised by four parameters, describing the size of the COP path over the support surface; the principal sway direction; and the shape and bandwidth of the power spectral density plot. COP one-dimensional time series (antero-posterior (AP) and medio-lateral (ML)) can be characterised by six parameters describing COP dispersion along the AP direction; mean velocity along the ML and AP directions; the contrast between ML and AP regulatory activity; and two parameters describing the spectral characteristics of the COP along the AP direction. On the basis of the results obtained, some guidelines are suggested for the choice of stabilometric parameters to use, with the aim of promoting standardisation in quantitative posturography.

The mitochondrial oxoglutarate carrier: cysteine-scanning mutagenesis of transmembrane domain IV and sensitivity of Cys mutants to sulfhydryl reagents.

Using a functional mitochondrial oxoglutarate carrier mutant devoid of Cys residues (C-less carrier), each amino acid residue in transmembrane domain IV and flanking hydrophilic loops (from T179 to S205) was replaced individually with Cys. The great majority of the 27 mutants exhibited significant oxoglutarate transport in reconstituted liposomes as compared to the activity of the C-less carrier. In contrast, Cys substitution for G183, R190, Q198, and Y202, in either C-less or wild-type carriers, yielded molecules with complete loss of oxoglutarate transport activity. G183 and R190 could be partially replaced only by Ala and Lys, respectively, whereas Q198 and Y202 were irreplaceable with respect to oxoglutarate transport. Of the single-Cys mutants tested, only T187C, A191C, V194C, and N195C were strongly inactivated by N-ethylmaleimide and by low concentrations of methanethiosulfonate derivatives. Oxoglutarate protects Cys residues at positions 187, 191, and 194 against reaction with N-ethylmaleimide. These positions as well as the residues found to be essential for the carrier activity, except Y202 which is located in the extramembrane loop IV-V, reside on the same face of transmembrane helix IV, probably lining part of a water-accessible crevice or channel between helices of the oxoglutarate carrier.

Border-tracing algorithm implementation for the femoral geometry reconstruction.

In some orthopaedic applications such as the design of custom-made hip prostheses, reconstruction of the bone morphology is a fundamental step. Different methods are available to extract the geometry of the femoral medullary canal from computed tomography (CT) images. In this research, an automatic procedure (border-tracing method) for the extraction of bone contours was implemented and validated. A composite replica of the human femur was scanned and the CT images processed using three different methods, a manual procedure; the border-tracing algorithm; and a threshold-based method. The resulting contours were used to estimate the accuracy of the implemented procedure. The two software techniques were more accurate than the manual procedure. Then, these two procedures were applied to an in vivo CT data set in order to determine to most critical region for repeatability. Only for the images located in this region, the repeatability measurement was carried out for six in vivo CT data sets to evaluate the inter-femur repeatability. The border-tracing method was found to achieve the highest repeatability.

HIDE: a new hybrid environment for the design of custom-made hip prosthesis.

This technical note describes a new software environment (HIPCOM design environment, HIDE) for the design of custom-made total hip replacements. These devices are frequently designed using general-purpose mechanical computer-aided design (CAD) programs using a set of bone contours extracted from the computer tomography (CT) images as anatomical reference. On the contrary, the HIDE system was developed to let the operator directly design the stem shape onto the CT images in a single-step operation. The operator can directly import CT data in DICOM format or use special functions to reconvert to a digital stack, the CT images printed on a radiological film. Once the stack of CT images is loaded, the operator can design the implant shape by imposing control sections directly on the CT images. The interpolation of these control sections produces the basic 3D shape of the custom-made stem. The shape is then exported to the CAD-computer-aided manufacturing (CAM) program to refine the design and to generate the part program to manufacture the implant with a CNC tooling machine. Using HIDE, the duration of design steps it affected was reduced by more than 50% with respect to the standard method in use at the manufacturer site. HIDE also improved the accuracy and the repeatability of the whole procedure. The learning curve became flat after only ten cases. These good results were achieved because of the integration of the vectorial description of the prosthetic component with the raster description of the CT data that allowed the designer to use all details available in the CT images.