Tumour-specific hybrid nanoparticles in therapy of breast cancer.

In this study, salicylic acid (SA) dopped into poly(3-hydroxybutyrate) (PHB) and prepared nanoparticles (NPs) to increase encapsulation efficiency, anti-cancer activity of caffeic acid (Caff), and folic acid (FA) for breast cancer treatment. NPs were prepared by solvent evaporation method and characterised by FTIR, DSC, SEM, and entrapment-loading efficiencies. The mean diameter, polydispersity index (PDI), and zeta potential (ZP) were evaluated by dynamic light scattering (DLS). In vitro release and stability studies were done via eppendorf method. The cytotoxicity, cell dead and internalisation of NPs were shown by MTT, fluorescein and confocal microscopy. The diameter and ZP of NPs were 172 ± 7 nm and -29 ± 0.38 mV. The entrapment efficiencies of 5 and 10 Caff NPs were 79 ± 0.23% and 70 ± 0.42%. NPs showed good stability within 30 d and sustained release over 25 d. FA-5Caff NPs showed 37 ± 0.3% viability on MCF-7. FA-Caff NPs were identified as promising carrier system for breast cancer therapy.

Three-dimensional MRI-based treatment planning approach for non-invasive ocular proton therapy.

PURPOSE: To develop a high-resolution three-dimensional (3D) magnetic resonance imaging (MRI)-based treatment planning approach for uveal melanomas (UM) in proton therapy. MATERIALS/METHODS: For eight patients with UM, a segmentation of the gross tumor volume (GTV) and organs-at-risk (OARs) was performed on T1- and T2-weighted 7 Tesla MRI image data to reconstruct the patient MR-eye. An extended contour was defined with a 2.5-mm isotropic margin derived from the GTV. A broad beam algorithm, which we have called πDose, was implemented to calculate relative proton absorbed doses to the ipsilateral OARs. Clinically favorable gazing angles of the treated eye were assessed by calculating a global weighted-sum objective function, which set penalties for OARs and extreme gazing angles. An optimizer, which we have named OPT'im-Eye-Tool, was developed to tune the parameters of the functions for sparing critical-OARs. RESULTS: In total, 441 gazing angles were simulated for every patient. Target coverage including margins was achieved in all the cases (V95%  > 95%). Over the whole gazing angles solutions space, maximum dose (Dmax ) to the optic nerve and the macula, and mean doses (Dmean ) to the lens, the ciliary body and the sclera were calculated. A forward optimization was applied by OPT'im-Eye-Tool in three different prioritizations: iso-weighted, optic nerve prioritized, and macula prioritized. In each, the function values were depicted in a selection tool to select the optimal gazing angle(s). For example, patient 4 had a T2 equatorial tumor. The optimization applied for the straight gazing angle resulted in objective function values of 0.46 (iso-weighted situation), 0.90 (optic nerve prioritization) and 0.08 (macula prioritization) demonstrating the impact of that angle in different clinical approaches. CONCLUSIONS: The feasibility and suitability of a 3D MRI-based treatment planning approach have been successfully tested on a cohort of eight patients diagnosed with UM. Moreover, a gaze-angle trade-off dose optimization with respect to OARs sparing has been developed. Further validation of the whole treatment process is the next step in the goal to achieve both a non-invasive and a personalized proton therapy treatment.

Evaluation of electrocardiographic ventricular repolarization parameters in extreme obesity.

BACKGROUND AND OBJECTIVES: The risk of sudden death and cardiac arrhythmia increases in morbidly obese patients. We aimed to evaluate the marker of arrhythmias such as Tp-e/QT, Tp-e/QTc, Tp-e/JT and Tp-e/JTc ratios in extreme obesity. METHODS: The study included 41 extremely obese patients and 41 control subjects. QTmax, QTmin, QRS, JT and Tp-e intervals were measured od 12­lead electrocardiographies. In addition, Tp-e/QT, Tp-e/QTc, Tp-e/JT and Tp-e/JTc rates and QTc, cQTd and JTc intervals were calculated. RESULTS: Tp-e interval (79.2 ±â€¯9.7 ms (milisecond) vs. 68.6 ±â€¯8.1, p < 0.001), QTc interval (395.9 ±â€¯18.8 vs. 377.9 ±â€¯19.3 ms, p < 0.001), JTc interval (317.1 ±â€¯27.0 vs. 297.4 ±â€¯23.2 ms, p = 0.001), Tp-e/QT ratio (0.22 ±â€¯0.03vs. 0.19 ±â€¯0.02, p < 0.001), Tp-e/QTc ratio (0.20 ±â€¯0.26vs. 0.18 ±â€¯0.02, p = 0.001), Tp-e/JT ratio (0.29 ±â€¯0.04 vs. 0.25 ±â€¯0.03, p < 0.001), TPe/JTc ratio (0.25 ±â€¯0.04 vs. 0.23 ±â€¯0.03, p = 0.018), QTd (32.8 ±â€¯10 vs.15 ±â€¯6.4 ms, p < 0.001) and cQTd (70.0 ±â€¯30.1 vs. 31.3 ±â€¯22.4 ms, p < 0.001) were significantly higher in obese patients. CONCLUSION: Compared to healthy subjects potential ECG repolarization predictors were significantly increased in extremely obese patients.

Evaluation of death pathway genes FAS and FASL polymorphisms in chronic HBV infection.

This study was designed to determine the possible asssociation between selected FAS and FASLG polymorphisms and Hepatitis B virus (HBV) infection. FAS-670 G/A, FAS-1377 G/A, FASLG-844 T/C and FASLG IVS2nt-124 A/G polymorphisms were genotyped by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). A total of age and sex matched 108 controls and a hundred chronic HBV patients were recruited to conduct a case-control study. FAS-670 polymorphism was associated with chronic HBV infection (P = 0.03) FAS-1377 GG, GA and AA genotypes among the cases (90%, 5% and 5%, respectively) were significantly different from those among the controls (68%, 31.5% and 5.6%; P = 0.00). FASLG-844 allele distribution was similar between the groups (P = 0.17) but TC genotype (67.3%) was frequent in chronic HBV patients, while CC genotype was found significantly higher (29.6%) in controls. No association between FASLG IVS2nt-124 polymorphism and chronic HBV infection could be identified (P = 0.55). FAS-670 polymorphism is associated with chronic HBV infection, while FASLG IVS2nt-124 A/G polymorphism is not. The FAS-1377G/A and FASLG-844 T/C genotypes are likely to play a substantial role in HBV infection. Further studies evaluating polymorphisms in other genes related with apoptosis are needed to elucidate the role of genetic variation in HBV infection.

Mutation analysis of the Vangl2 coding region revealed no common cause for Tetralogy of Fallot.

Vangl2 (Van Gogh-like 2) protein acts via non-canonical Wnt signalling to regulate polarized cell movements during development of the proximal outflow tract in vertebrate embryos. Recently, it has been shown that mutations of the Vangl2 gene cause aortic arch defects that are characteristic of the loop-tail (Lp) mouse and they have also became a strong candidate for causing congenital outflow tract defects in humans. Thus, in this study Tetralogy of Fallot (ToF), which comprises a group of syndromes that constitutes the most frequent cause of congenital cardiac outflow abnormalities in humans, was analysed for mutations within all coding regions of the Vangl2 gene. Based on direct sequencing data from a combination of 20 patients with ToF and 22 healthy people, three polymorphisms have been identified in exon 6 and exon 7 which do not change the amino acid sequence. It was concluded, therefore, that there is no specific mutation responsible for the ToF phenotype in the Vangl2 gene.

Evaluation of glucose metabolism and reproductive hormones in polycystic ovary syndrome on the basis of peroxisome proliferator-activated receptor (PPAR)-gamma2 Pro12Ala genotype.

BACKGROUND: Peroxisome proliferator-activated receptor (PPAR)-gamma2 Pro12Ala polymorphism has been suggested as a protective factor for polycystic ovary syndrome (PCOS). In this study, we aimed to investigate metabolic features and reproductive hormones in women with PCOS and compare these features with control women on the basis of Pro12Ala genotype. METHODS: This study involved 60 randomly selected women with PCOS and 60 controls. Main outcome measures were anthropometric measures, variables of glucose metabolism and reproductive hormones. All the patients were genotyped for Pro12Ala variant of PPAR-gamma2 gene. RESULTS: Patients with Pro12Ala polymorphism were more obese in both groups. Furthermore, they had lower fasting insulin levels, were less insulin-resistant and were less glucose-intolerant as demonstrated by 2 h glucose concentrations. However, there was no difference in reproductive hormone levels on the basis of Pro12Ala genotype. CONCLUSIONS: Both control women and women with PCOS had significant differences in glucose metabolism on the basis of PPAR-gamma2 Pro12Ala polymorphism. Pro12Ala variant may break the process that leads to PCOS in susceptible women, instead of being a direct causal relationship between Pro12Ala polymorphism and PCOS.

The importance of IRS-1 Gly972Arg polymorphism in evaluating the response to metformin treatment in polycystic ovary syndrome.

BACKGROUND: Recent evidence suggests that one of the modes of action of metformin may be through phosphorylation of the insulin receptor and insulin receptor substrates. With this in mind, we supposed that the G972A variant of insulin receptor substrate-1 (IRS-1) may modulate the response to metformin treatment in women with polycystic ovary syndrome (PCOS). METHODS: This preliminary study involved 60 randomly selected women with PCOS. All patients received dietary instructions and metformin 500 mg three times daily for 6 months. Main outcome measures were androgen levels, parameters of glucose and insulin metabolism and anthropometric variables. After a second evaluation of the patients at 6 months, they were genotyped for the Gly972Arg variant of the IRS-1 gene. RESULTS: Metformin had differential effects on fasting insulin levels, insulin resistance as demonstrated by homeostasis model assessment (HOMA), LH, total testosterone, dehydroepiandrosterone sulphate and free testosterone index on the basis of IRS genotype. The response to metformin therapy in other parameters was not different according to IRS genotype. CONCLUSION: There was a differential effect of metformin therapy in PCOS women on the basis of IRS genotype. This study may call attention to the importance of molecular markers in the management of women with PCOS.

Association of T102C polymorphism of the 5-HT2A receptor gene with psychiatric status in fibromyalgia syndrome.

Serotonin (5-HT) is a key neurotransmitter in the central nervous system. It is suggested that serotonergic dysfunction may be involved in the pathophysiology of fibromyalgia syndrome (FS). In this study, we aimed to investigate T102C polymorphism of the 5-HT2A receptor gene in FS. Fifty-eight patients with FS and 58 unrelated healthy volunteer controls were included in the study. In both groups, the C/C, C/T, and T/T genotypes of the 5-HT gene were represented in 31% (22.4% in controls), 50% (53.4%), and 19% (24.1%), respectively. The 5-HT2A receptor gene polymorphism results were not significantly different between patients and controls (chi squared test, P>0.05). There was a significant correlation between patients with the T/T genotype and the subgroup according to the SCL-90-R test, (analysis of variance, P<0.05). We also saw that patients with the T/T genotype had the lowest pain threshold. CONCLUSION. T102C polymorphism of the 5-HT2A receptor gene is not associated with the etiology of FS. Our results also indicate that the T/T genotype may be responsible for psychiatric symptoms of FS.

Possible association of temporomandibular joint pain and dysfunction with a polymorphism in the serotonin transporter gene.

The purpose of this study was to evaluate the relationship between temporomandibular joint pain and dysfunction and serotonin transporter (5-HTT) gene polymorphism. Forty-eight patients with temporomandibular joint pain and 111 healthy control subjects were examined. The results for the patients and control subjects were not significantly different (P >.05). The analysis of genotype distribution (homozygous for STin 2.10 genotypes of the variable-number tandem-repeat polymorphism) showed significant differences between the patients and control subjects (P =.003). ST 2.10 allele was more frequent in the patients with temporomandibular joint pain and dysfunction. In the control group, however, STin 2.12/12 genotype was significantly higher (P =.017). In the patients who were homozygous or heterozygous for variable-number tandem-repeat variants of 5-HTT STin 2.12 copies, the average scores of somatization and anger were significantly higher than those who were homozygous for STin 2.10 variant (P <.05). The patients who were homozygous for STin 2.10 genotype were also homozygous for "L" genotype (P =.019). However, this was not the condition in the control subjects. This study does not provide evidence to support the involvement of 5-HTT gene-linked polymorphic region in temporomandibular joint pain and dysfunction. Our findings indicated that only the presence of the homozygous STin 2.10 genotype of variable-number tandem-repeat is likely to play a substantial role in the genetic predisposition to temporomandibular joint pain and dysfunction and that the STin 2.12/12 genotype may have a protective role against temporomandibular joint pain and dysfunction.

Processing of tetanus and botulinum A neurotoxins in isolated chromaffin cells.

Tetanus and botulinum A neurotoxins were introduced into the cytosol of chromaffin cells by means of an electric field in which the plasma membrane is forced to form pores of approximately 1 micron at the sites facing the electrodes. As demonstrated by electron microscopy, both [125I] and gold-labelled tetanus toxin (TeTx) diffuse through these transient openings. Dichain-TeTx, with its light chain linked to the heavy chain by means of a disulfide bond, causes the block of exocytosis to develop more slowly than does the purified light chain. The disulfide bonds, which in both toxins hold the subunits together, were cleaved by the intrinsic thioredoxin-reductase system. Single chain TeTx, in which the heavy and light chains are interconnected by an additional peptide bond, was far less effective than dichain-TeTx at blocking exocytosis, which indicates that proteolysis is the rate-limiting step. The toxins were degraded further to low-molecular weight fragments which, together with intact toxins and subunits, were released by the cells. The intracellular half-life of [125I] dichain-TeTx was approximately three days. The number of light-chain molecules required to maintain exocytosis block in a single cell, as calculated by two different methods, was less than 10. The long duration of tetanus poisoning may result from the persistence of intracellular toxin due to scarcity of free cytosolic proteases. This may also hold for the slow recovery from botulism.