Clinical and metabolic implications of obesity in prostate cancer: is testosterone a missing link?

Objectives: To assess sex hormones in men with obesity and prostate cancer (PCa) and to study association between androgens and the pathogenesis biology of PCa in vitro. Subjects and methods: One hundred and eighty-one men older than 45 years selected from of a population attending to Urology departments screening for PCa, (78 participants without PCa and 103 patients with PCa). All participants were assessed for body mass index (BMI), age, Gleason score, and PSA. Endocrine profile was determined for LH, total testosterone (TT), 17ß-estradiol (E2), prolactin and leptin. Biochemical profile (HbA1c, triacylglycerols and lipoproteins) was also determined. In vitro experiments were also performed, involving the study of 5α-dihydrotestosterone (DHT) and E2 in the presence of adipocyte-conditioned medium (aCM). Results: All variables were continuous and described a Gaussian distribution unless mentioned. To determine the relation of aggressiveness, variable were transformed into categories. Thus, PCa aggressiveness is associated with the increase of age and BMI (p < .0001) but with is decreased with TT and E2 (p < .05). Moreover, adipocyte-secreted molecules increase aggressiveness of PCa cells in vitro. Lastly, DTH but not E2 enables invasiveness in vitro. Conclusions: It was observed a coexistence of hormone axis profile alteration with sex hormones and BMI in PCa patients, in accordance with the new perspective of PCa pathogenesis.

Intestinal anti-inflammatory effects of RGD-functionalized silk fibroin nanoparticles in trinitrobenzenesulfonic acid-induced experimental colitis in rats.

BACKGROUND: Current treatment of inflammatory bowel disease is based on the use of immunosuppressants or anti-inflammatory drugs, which are characterized by important side effects that can limit their use. Previous research has been performed by administering these drugs as nanoparticles that target the ulcerated intestinal regions and increase their bioavailability. It has been reported that silk fibroin can act as a drug carrier and shows anti-inflammatory properties. PURPOSE: This study was designed to enhance the interaction of the silk fibroin nanoparticles (SFNs) with the injured intestinal tissue by functionalizing them with the peptide motif RGD (arginine-glycine-aspartic acid) and to evaluate the intestinal anti-inflammatory properties of these RGD-functionalized silk fibroin nanoparticles (RGD-SFNs) in the trinitrobenzenesulfonic acid (TNBS) model of rat colitis. MATERIALS AND METHODS: SFNs were prepared by nanoprecipitation in methanol, and the linear RGD peptide was linked to SFNs using glutaraldehyde as the crosslinker. The SFNs (1 mg/rat) and RGD-SFNs (1 mg/rat) were administered intrarectally to TNBS-induced colitic rats for 7 days. RESULTS: The SFN treatments ameliorated the colonic damage, reduced neutrophil infiltration, and improved the compromised oxidative status of the colon. However, only the rats treated with RGD-SFNs showed a significant reduction in the expression of different pro-inflammatory cytokines (interleukin [IL]-1ß, IL-6, and IL-12) and inducible nitric oxide synthase in comparison with the TNBS control group. Moreover, the expression of both cytokine-induced neutrophil chemoattractant-1 and monocyte chemotactic protein-1 was significantly diminished by the RGD-SFN treatment. However, both treatments improved the intestinal wall integrity by increasing the gene expression of some of its markers (trefoil factor-3 and mucins). CONCLUSION: SFNs displayed intestinal anti-inflammatory properties in the TNBS model of colitis in rats, which were improved by functionalization with the RGD peptide.

The impact of a specific blend of essential oil components and sodium butyrate in feed on growth performance and Salmonella counts in experimentally challenged broilers.

Essential oils (EO) and short-chain fatty acids have potential antimicrobial activity in broilers. This study aimed to investigate the effect of a specific blend of EO and a combination of this blend of EO with sodium-butyrate on growth performance and Salmonella colonization in broilers. A total of 480 one-day-old male broilers were distributed into 5 treatments (8 pens per treatment and 12 birds per pen) and reared during 42 d in experimental conditions. Dietary treatments consisted of the addition of different doses of EO (0 mg/kg, control; 50 mg/kg, EO50 and 100 mg/kg, EO100) or a combination of EO with 1 g/kg of sodium-butyrate (B; EO50 + B, EOB50 and EO100 + B, EOB100) to a basal diet. All birds were orally infected with 10(8) cfu of Salmonella Enteritidis on d 7 of study. Individual BW and feed intake per pen were measured at arrival and on a weekly basis. The prevalence and enumeration of Salmonella in feces was determined per treatment at 72 h postinfection and on d 23 and 37 of study. At slaughter, cecal content and liver samples from 16 birds per treatment were cultured for Salmonella and cecal pH was measured. No differences were observed on growth performance among treatments. All fecal samples analyzed were positive for Salmonella from d 10 to the end of the rearing period. At slaughter, Salmonella contamination (positive samples) in cecum was lower in birds fed EOB50 compared with the other treatments (P < 0.05), whereas birds fed the control diet showed the highest colonization rates. The pH of the cecal content was not different among treatments. Thus, EO or its combination with sodium-butyrate did not affect growth performance. However, a clear effectiveness of these products was observed in Salmonella control, especially when low doses of EO were combined with sodium-butyrate (EOB50).

A 100 kV, 60 A solid state 4 kHz switching modulator for high power klystron driving.

A solid state high power modulator capable of delivering 120 kV and 60 A developed in collaboration with the JEMA Corporation, ESS Bilbao, and the SNS (ORNL) for driving high power klystrons is presented. Pulses with less than 10 µs risetime and flatness under 0.1% are obtained with programmable frequency pulses between 2 and 50 Hz. Eight solid state switches combined with custom air-insulated high voltage transformers working at a switching frequency of 4 kHz produce high quality pulses by phase shifting the transformer drives. Each relative high frequency stage pumps a double stage high voltage Marx generator that supplies the output pulse shape and frequency. This merged topology between a Marx generator and direct modulator takes advantage of the strengths of both approaches. Low energy storage in the output stages assures safe operation in case of a load arc discharge. Real time voltage correction during the pulse is also provided to compensate for the droop inherent with the use of low energy storage in the output stages. Data at full power with a dummy resistive load are presented.

The fate of nanocarriers as nanomedicines in vivo: important considerations and biological barriers to overcome.

Many pharmaceuticals on the market suffer from two significant limitations to their activity: lack of specificity toward the pathological site and poor aqueous solubility. Both factors therefore require the application of a large total dose of a drug to achieve high local concentration, causing numerous off-target toxic effects. Consequently, the grand aim of targeted drug delivery - the often-referred "magic bullet" - promises to improve drug concentration at the target site and maximize therapeutic response. Nanomaterial drug delivery systems have been explored extensively in the recent years for just this purpose. In the field of medicine, nanocarriers (NCs) have the potential to improve the biodistribution and pharmacokinetic characteristics of drugs, thereby reducing side effects while improving the therapeutic effect of drugs. Many nanomaterials are exquisitely designed and possess potent properties, yet it is extremely important to note that a general understanding of the interaction of nanomaterials with biological systems is essential for any such model properties to be effective in vivo, since the body presents a host of biological 'barriers' that will be encountered drug NCs. This review offers a general overview of the different biological obstacles that a NC must negotiate before it can carry out its desired role as a medicinal agent. From this standpoint we suggest aspects that should be considered for the rational design of novel nanomaterials possessing physicochemical properties that are appropriate for therapeutic or theragnostic applications.

Functionalized Fe3O4@Au superparamagnetic nanoparticles: in vitro bioactivity.

The interaction of nanoparticles with cells has been a focus of interest during the past decade. We report the fabrication and characterization of hydrosoluble Fe3O4@Au nanoparticles functionalized with biocompatible and fluorescent molecules and their interaction with cell cultures by visualizing them with confocal microscopy. Gold covered iron oxide nanoparticles were synthesized by reducing metal salts in the presence of oleylamine and oleic acid. The functionalization of these particles with an amphiphilic polymer provides a water soluble corona as well as the possibility to incorporate different molecules relevant for bio-applications such as poly(ethylene glycol), glucose or a cadaverine derived dye. The particle size, and the presence of polymer layers and conjugated molecules were characterized and confirmed by transmission electron microscopy, thermogravimetric measurements and infrared spectroscopy. A complete magnetic study was performed, showing that gold provides an optimum coating, which enhances the superparamagnetic behaviour observed above 10-15 K in this kind of nanoparticle. The interaction with cells and the cytotoxicity of the Fe3O4@Au preparations were determined upon incubation with the HeLa cell line. These nanoparticles showed no cytotoxicity when evaluated by the MTT assay and it was demonstrated that nanoparticles clearly interacted with the cells, showing a higher level of accumulation in the cells for glucose conjugated nanoparticles.

A ceramic microreactor for the synthesis of water soluble CdS and CdS/ZnS nanocrystals with on-line optical characterization.

In this paper, a computer controlled microreactor to synthesize water soluble CdS and CdS/ZnS nanocrystals with in situ monitoring of the reaction progress is developed. It is based on ceramic tapes and the Low-Temperature Co-fired Ceramics technology (LTCC). As well the microsystem set-up, the microreactor fluidic design has also been thoroughly optimized. The final device is based on a hydrodynamic focusing of the reagents followed by a three-dimensional micromixer. This generates monodispersed and stable CdS and core-shell CdS/ZnS nanocrystals of 4.5 and 4.2 nm, respectively, with reproducible optical properties in terms of fluorescence emission wavelengths, bandwidth, and quantum yields, which is a key requirement for their future analytical applications. The synthetic process is also controlled in real time with the integration of an optical detection system for absorbance and fluorescence measurements based on commercial miniaturized optical components. This makes possible the efficient managing of the hydrodynamic variables to obtain the desired colloidal suspension. As a result, a simple, economic, robust and portable microsystem for the well controlled synthesis of CdS and CdS/ZnS nanocrystals is presented. Moreover, the reaction takes place in aqueous medium, thus allowing the direct modular integration of this microreactor in specific analytical microsystems, which require the use of such quantum dots as labels.

In vitro transcription and translation inhibition via DNA functionalized gold nanoparticles.

The use of gold nanoparticles (AuNPs) has been gaining momentum as vectors for gene silencing strategies, combining the AuNPs' ease of functionalization with DNA and/or siRNA, high loading capacity and fast uptake by target cells. Here, we used AuNP functionalized with thiolated oligonucleotides to specifically inhibit transcription in vitro, demonstrating the synergetic effect between AuNPs and a specific antisense sequence that blocks the T7 promoter region. Also, AuNPs efficiently protect the antisense oligonucleotide against nuclease degradation, which can thus retain its inhibitory potential. In addition, we demonstrate that AuNPs functionalized with a thiolated oligonucleotide complementary to the ribosome binding site and the start codon, effectively shut down in vitro translation. Together, these two approaches can provide for a simple yet robust experimental set up to test for efficient gene silencing of AuNP-DNA conjugates. What is more, these results show that appropriate functionalization of AuNPs can be used as a dual targeting approach to an enhanced control of gene expression-inhibition of both transcription and translation.

Nuclear localization of HIV-1 tat functionalized gold nanoparticles.

The impermeable nature of the cell plasma membrane limits the therapeutic uses of many macromolecules and there is therefore a growing effort to circumvent this problem by designing strategies for targeted intracellular delivery. During the last decade several cell penetrating peptides, such as the HIV-1 tat peptide, have been shown to traverse the cell membrane, where integral protein transduction domains (PTDs) are responsible for their cellular uptake, and to reach the nucleus while retaining biological activity. It has since been discovered that PTDs can enable the cellular delivery of conjugated biomolecules and even nanoparticles, but nuclear delivery has remained problematic. This present study focuses on the development of water soluble, biocompatible gold nanoparticles of differing size functionalized with the HIV-1 tat PTD with the aim of producing nuclear targeting agents. The particles were subsequently tested in vitro with a human fibroblast cell line, with results demonstrating successful nanoparticle transfer across the plasma membrane, with 5 nm particles achieving nuclear entry while larger 30 nm particles are retained in the cytoplasm, suggesting entry is blocked via nuclear pores dimensions.

Fe impurities weaken the ferromagnetic behavior in Au nanoparticles.

In this Letter, we report on a crucial experiment showing that magnetic impurities reduce the ferromagnetic order temperature in thiol-capped Au glyconanoparticles (GNPs). The spontaneous magnetization of AuFe GNPs exhibits a fast decrease with temperature that contrasts with the almost constant value of the magnetization observed in Au NPs. Moreover, hysteresis disappears below 300 K. Both features indicate that Fe impurities reduce the high local anisotropy field responsible for the ferromagnetic behavior in Au GNPs. As a consequence, the amazing ferromagnetism in Au NPs should not be associated with the presence of magnetic impurities.

Gold nanoparticles with different capping systems: an electronic and structural XAS analysis.

Gold nanoparticles (NPs) have been prepared with three different capping systems: a tetralkylammonium salt, an alkanethiol, and a thiol-derivatized neoglycoconjugate. Also gold NPs supported on a porous TiO(2) substrate have been investigated. X-ray absorption spectroscopy (XAS) has been used to determine the electronic behavior of the different capped/supported systems regarding the electron/hole density of d states. Surface and size effects, as well as the role of the microstructure, have been also studied through an exhaustive analysis of the EXAFS (extended X-ray absorption fine structure) data. Very small gold NPs functionalized with thiol-derivatized molecules show an increase in d-hole density at the gold site due to Au-S charge transfer. This effect is overcoming size effects (which lead to a slightly increase of the d-electron density) for high S:Au atomic ratios and core-shell microstructures where an atomically abrupt Au-S interface likely does not exist. It has been also shown that thiol functionalization of very small gold NPs is introducing a strong distortion as compared to fcc order. To the contrary, electron transfer from reduced support oxides to gold NPs can produce a higher increase in d-electron density at the gold site, as compared to naked gold clusters.

Permanent magnetism, magnetic anisotropy, and hysteresis of thiol-capped gold nanoparticles.

We report on the experimental observation of magnetic hysteresis up to room temperature in thiol-capped Au nanoparticles with 1.4 nm size. The coercive field ranges from 860 Oe at 5 K to 250 Oe at 300 K. It is estimated that the Au atoms exhibit a magnetic moment of mu=0.036mu(B). However, Au nanoparticles with similar size but stabilized by means of a surfactant, i.e., weak interaction between protective molecules and Au surface atoms, are diamagnetic, as bulk Au samples are. The apparent ferromagnetism is consequently associated with 5d localized holes generated through Au-S bonds. These holes give rise to localized magnetic moments that are frozen in due to the combination of the high spin-orbit coupling (1.5 eV) of gold and the symmetry reduction associated with two types of bonding: Au-Au and Au-S.

Sleep-EEG in borderline patients without concomitant major depression: a comparison with major depressives and normal control subjects.

The link between borderline personality disorder (BPD) and the affective disorders remains controversial. The aim of this study was to examine the relationships between BPD and major depression (MD) from the perspective of sleep parameters and to contribute to the characterisation of the sleep-EEG in BPD. We compared 20 off-medication BPD in-patients without co-existing MD with 20 sex- and age-matched MD patients without BPD and 20 sex- and age-matched control subjects. BPD patients had a greater prevalence of drug or alcohol abuse and suicide attempts than MD patients. MD patients had higher scores on the Hamilton Depression Rating Scale (HDRS). Both BPD and MD patients had less total sleep time, more prolonged sleep onset latency, and a greater percentage of wakefulness than control subjects. BPD patients and control subjects had more stage 2 sleep than MD patients. BPD patients had a longer duration of rapid eye movement (REM) sleep, and less stage 3, stage 4 and slow wave sleep than MD patients and control subjects. REM latency did not differentiate the three groups. BPD and MD patients shared sleep-continuity characteristics, but sleep architecture differentiated the two groups. BPD patients with a past history of MD had more wakefulness and less slow wave sleep than BPD patients without a history of MD; other sleep parameters, age, sex and HDRS scores were not statistically different in the two BPD subgroups. Although BPD and MD may coexist, the present study offers more arguments favouring the concept that they are not biologically linked and that BPD patients with depressive symptoms often experience an affective syndrome different from that in MD patients without BPD, in terms of quality and duration of symptoms and of the biological substrate.

Choline-binding domain as a novel affinity tag for purification of fusion proteins produced in Pichia pastoris.

The choline-binding domain (ChoBD) of the carboxy-terminal region of the Streptococcus pneumoniae amidase LYTA (C-LYTA) presents a strong affinity for tertiary amines. We report a method for single-step purification of proteins expressed in the methylotrophic yeast Pichia pastoris based on the fusion of C-LYTA to the protein of interest. We show that C-LYTA can be efficiently expressed and secreted in this host. Tagged proteins fused to this binding domain can be purified on inexpensive DEAE matrices. It therefore provides a useful system for the purification of recombinant proteins with high specificity suitable for industrial purposes.

Adhesion Forces between Lewis(X) Determinant Antigens as Measured by Atomic Force Microscopy.

The adhesion forces between individual molecules of Lewis(X) trisaccharide antigen (Le(X) ) have been measured in water and in calcium solution by using atomic force microscopy (AFM, see graph). These results demonstrate the self-recognition capability of this antigen, and reinforce the hypothesis that carbohydrate-carbohydrate interaction could be considered as the first step in the cell-adhesion process in nature.

Electroencephalographic abnormalities in borderline personality disorder.

Epilepsy and non-localized brain dysfunction have been invoked, among others, as underlying factors in borderline personality disorder. We have recorded 58 electroencephalograms in 20 borderline patients, first after complete drug washout and then under carbamazepine or placebo double-blind treatment. Taking into account only definite abnormal tracings, we found a 40% incidence of abnormal diffuse slow activity. No patient disclosed focal or epileptiform EEG features. Carbamazepine did not appear to modify the electroencephalogram.

Brain glucose metabolism in borderline personality disorder.

We searched for regional cerebral metabolic disturbances in patients with borderline personality disorder (BPD). Ten inpatients with BPD, no current DSM-IIIR Axis I diagnosis and free of any psychotropic substances, were compared with 15 age-matched control subjects using positron emission tomography with 2-deoxy-2-[18F]fluoro-D-glucose and semiquantitative analysis of regional glucose metabolic activity. We found relative hypometabolism in patients with borderline personality disorder at the level of the premotor and prefrontal cortical areas, the anterior part of the cingulate cortex and the thalamic, caudate and lenticular nuclei. This study shows significant cerebral metabolic disturbances in patients with borderline personality disorder. These metabolic disturbances, which are similar to some of those described in other psychiatric entities, may help to understand the characteristic clinical aspects of this disorder.

Aluminum tolerance in transgenic plants by alteration of citrate synthesis.

Aluminum when in soluble form, as found in acidic soils that comprise about 40 percent of the world's arable land, is toxic to many crops. Organic acid excretion has been correlated with aluminum tolerance in higher plants. Overproduction of citrate was shown to result in aluminum tolerance in transgenic tobacco (Nicotiana tabacum) and papaya (Carica papaya) plants.

TRH stimulation and dexamethasone suppression in borderline personality disorder.

The link between borderline personality disorder (BPD) and affective disorders is controversial. The dexamethasone suppression test (DST) and the thyrotropin-releasing hormone (TRH) stimulation test, which are useful in the study of affective illness, might help to elucidate this possible link. This report assessed these endocrine tests in a sample of 20 borderline patients without a concomitant diagnosis of major depression (but showing depressive symptoms) in comparison to a group of sex- and age-matched patients with major depressive disorder (MDD) without BPD. Only 5 of our BPD patients were DST nonsuppressors compared to 13 MDD patients at a threshold of 50 micrograms/L. With a threshold of delta max TSH < 5 microU/mL following TRH, 1 BPD patient showed a blunted TSH response compared to 9 MDD patients. BPD patients displayed significantly less perturbed tests. These results show no evidence of an endocrine biological link between BPD and the MDD. The depressive symptoms observed in BPD patients might not have the same biological substrates as those found in patients with MDD.