[Comparative analysis of antibiotic therapy and cytokine therapy (the drug Superlymph) for male infertility associated with male accessory gland infection].

AIM: To compare the effects of cytokine and antibacterial therapy on semen parameters and additional functional tests in patients with infertility due to male accessory gland infections (MAGI) who are preparing for assisted reproductive technologies (ART). MATERIALS AND METHODS: A randomized, prospective, controlled clinical trial without blinding was carried out. A total of 60 men from infertile couples with MAGI who were preparing to ART was included in the study. In the main group (n=30) patients received Superlymph, 1 suppository of 25 units, 2 times a day for 20 days. In the control group (n=30), the antibacterial drug Doxycycline 100 mg 2 times a day for 28 days was given. After the end of therapy on day 33+/-3, a repeated sperm analysis, MAR test, a test for reactive oxygen species in the ejaculate and sperm DNA fragmentation, and bacteriological examination of sperm was performed. In addition, a survey for adverse events was carried out. The significance of differences in initial and final parameters and differences between groups was assessed using the Students t-test, Wilcoxon test and Mann-Whitney U-test depending on the data distribution. The Shapiro-Wilk test was done to investigate the normality of data distribution. Fisher's exact test was used to compare categorical variables. RESULTS: The final analysis included data from 53 patients (n=28 in the main group and n=25 in the control group). In both groups, a significant decrease in the level of free oxygen radicals in the ejaculate was noted (p=0.031), which was more pronounced in the main group. There were no differences in other semen parameters. Eradication of the microorganism according to bacteriological examination occurred in 57.1% of patients in the main group and in 88% of those in the control group (p=0.016). In patients receiving monotherapy with Superlymph, there was a significant decrease in the sperm DNA fragmentation index and the concentration of leukocytes in the ejaculate. In patients receiving antibacterial therapy, there was a significant increase in ejaculate volume, a decrease in the proportion of IgG-associated sperm and leukocyte concentration. CONCLUSION: Many issues of diagnosis and treatment of MAGI have not been thoroughly studied and are poorly standardized. Considering the problems of increasing antibiotic resistance, alternative treatment options are needed. Cytokine therapy (the drug Superlymph) is an effective alternative method of monotherapy for male infertility due to MAGI and is optimal for quickly preparing a couple for ART protocol, given its positive effect on oxidative stress and the index of sperm DNA fragmentation. The prospect of combination therapy with antibiotics and Superlymph seems promising.

Ion beam figuring with focused anode layer thruster.

This work presents the peculiarities of cone ion beam formation with a focused thruster with anode layer (TAL) and its application to silicon carbide (SiC) ion beam figuring. Modeling results of Lorentz E × B force distribution in the discharge gap are presented. 3D particle tracing for keV Ar ions is carried out for the first time in the beam drift region of TAL with magnetic lens. Extracted ion beam full width at half maxima is about 2 mm in the focal plane, where the SiC etching rate reaches 0.5 µm/min. The SiC sputter yields are measured as a function of the Ar ion impact energy and beam incidence angle. The maximum sputter yield of 2.8 atom/ion is observed at 45° of the beam-sample angle for the Si targets. Furthermore, the maximum sputter yield value of 1.7 atom/ion is measured at 30° of the beam-sample angle for the SiC targets. The novelty of present research is in the application of focused TAL keV Ar ion beam to the SiC ion beam figuring.

High resolution Thomson scattering system for steady-state linear plasma sources.

The high resolution Thomson scattering system with 63 points along a 25 mm line measures the radial electron temperature (Te) and its density (ne) in an argon plasma. By using a DC arc source with lanthanum hexaboride (LaB6) electrode, plasmas with electron temperature of over 5 eV and densities of 1.5 × 1019 m-3 have been measured. The system uses a frequency doubled (532 nm) Nd:YAG laser with 0.25 J/pulse at 20 Hz. The scattered light is collected and sent to a triple-grating spectrometer via optical-fibers, where images are recorded by an intensified charge coupled device (ICCD) camera. Although excellent in stray-light reduction, a disadvantage comes with its relatively low optical transmission and in sampling a tiny scattering volume. Thus requires accumulating multitude of images. In order to improve photon statistics, pixel binning in the ICCD camera as well as enlarging the intermediate slit-width inside the triple-grating spectrometer has been exploited. In addition, the ICCD camera capture images at 40 Hz while the laser is at 20 Hz. This operation mode allows us to alternate between background and scattering shot images. By image subtraction, influences from the plasma background are effectively taken out. Maximum likelihood estimation that uses a parameter sweep finds best fitting parameters Te and ne with the incoherent scattering spectrum.

Role of transient receptor potential vanilloid 1 in regulating erythropoietin-induced activation of endothelial nitric oxide synthase.

AIMS: Erythropoietin (EPO), the key hormone involved in erythropoiesis, beneficially affects endothelial cells (ECs), but the detailed mechanisms are yet to be completely understood. In this study, we investigated the role of transient receptor potential vanilloid type 1 (TRPV1), a ligand-gated non-selective calcium (Ca2+ ) channel, in EPO-mediated endothelial nitric oxide synthase (eNOS) activation and angiogenesis. METHODS AND RESULTS: In ECs, EPO time dependently increased intracellular levels of calcium; this increase was abrogated by the Ca2+ chelators and pharmacological inhibitors of TRPV1 in bovine aortic ECs (BAECs) and TRPV1-transfected HEK293 cells. In addition, EPO-induced nitrite oxide (NO) production, phosphorylation of eNOS, Akt and AMP-activated protein kinase (AMPK) and the formation of TRPV1-Akt-AMPK-eNOS complex as well as tube formation were diminished by the pharmacological inhibition of TRPV1 in BAECs. Moreover, EPO time dependently induced the phosphorylation of phospholipase C-γ1 (PLC-γ1). Inhibition of PLC-γ1 activity blunted the EPO-induced Ca2+ influx, eNOS phosphorylation, TRPV1-eNOS complex formation and NO production. The phosphorylated level of eNOS increased in the aortas of EPO-treated wild-type (WT) mice or EPO-transgenic (Tg) mice but not in those of EPO-treated TRPV1-deficient (TRPV1-/- ) mice or EPO-Tg/TRPV1-/- mice. Matrigel plug assay showed that EPO-induced angiogenesis was abrogated in TRPV1 antagonist capsazepine-treated WT mice and TRPV1-/- mice. CONCLUSION: These findings indicate the EPO-induced Ca2+ influx via the activation of the PLC-γ1 signalling pathway, which leads to TRPV1 activation and consequently increases the association of the TRPV1-Akt-AMPK-eNOS complex, eNOS activation, NO production and angiogenesis.

The essential role of transient receptor potential vanilloid 1 in simvastatin-induced activation of endothelial nitric oxide synthase and angiogenesis.

AIMS: We investigated the role of transient receptor potential vanilloid receptor type 1 (TRPV1) in simvastatin-mediated activation of endothelial nitric oxide synthase (eNOS) and angiogenesis. METHODS: Fluo-8 NW assay was for Ca(2+) detection; Griess's assay was for NO bioavailability; Western blotting and immunoprecipitation were for protein phosphorylation and interaction; tube formation and Matrigel plug assay were for angiogenesis. RESULTS: In endothelial cells (ECs), treatment with simvastatin time-dependently increased intracellular level of Ca(2+). Pharmacological inhibition or genetic disruption of TRPV1 abrogated simvastatin-mediated elevation of intracellular Ca(2+) in ECs or TRPV1-transfected HEK293 cells. Loss of TRPV1 function abolished simvastatin-induced NO production and phosphorylation of eNOS and calmodulin protein kinase II (CaMKII) in ECs and in aortas of mice. Inhibition of TRPV1 activation prevented the simvastatin-elicited increase in the formation of TRPV1-Akt-CaMKII-AMPK-eNOS complex. In mice, Matrigel plug assay showed that simvastatin-evoked angiogenesis was abolished by TRPV1 antagonist and genetic ablation of TRPV1. Additionally, our results demonstrated that TRP ankyrin 1 (TRPA1) is the downstream effector in the simvastatin-activated TRPV1-Ca(2+) signalling and in the consequent NO production and angiogenesis as evidence by that re-expression of TRPA1 further augmented simvastatin-elicited Ca(2+) influx in TRPV1-expressed HEK293 cells and ablation of TRPA1 function profoundly inhibited the simvastatin-induced increase in the phosphorylation of eNOS and CaMKII, formation of TRPV1-Akt-CaMKII-AMPK-eNOS complex, NO bioavailability, tube formation and angiogenesis in ECs or mice. CONCLUSION: Simvastatin-induced Ca(2+) influx may through the activation of TRPV1-TRPA1 signalling, which leads to phosphorylation of CaMKII, increases in the formation of TRPV1-CaMKII-AMPK-eNOS complex, eNOS activation, NO production and, ultimately, angiogenesis in ECs.

Factors predictive of perforation during endoscopic submucosal dissection for the treatment of colorectal tumors.

BACKGROUND AND AIM: Although perforation of the colon is known as one of the main complications of endoscopic submucosal dissection (ESD) for colorectal tumor management, factors predictive of perforation have not been fully evaluated. This study aimed to determine the factors associated with perforation during colorectal ESD. METHODS: Patients with colorectal tumors undergoing ESD were enrolled and their records were reviewed retrospectively. Age, sex, co-morbidity, medication history, procedure time, resection method, tumor size, location, gross morphology, the presence of fibrosis, and histologic findings were included as possible risk factors. In the cases where perforation had occurred, factors associated with the duration of hospitalization were analyzed. RESULTS: One hundred eight lesions in 108 patients were eligible for inclusion in the study (68 patients were male; mean patient age was 63.01 ± 10.71 years). Mean tumor size was 27.59 ± 10.10 mm (range: 8 - 53 mm). Laterally spreading tumor was the most common type (75 %), followed by the protruding type (25 %). Procedure time was 61.95 ± 41.90 minutes (range: 5 - 198 minutes). Complete en bloc resection was achieved for 85 lesions (78.7 %). Perforation occurred in 22 patients (20.4 %). Multivariate analysis confirmed that tumor size [odds ratio (OR): 1.084; 95 % confidence interval (CI): 1.015 - 1.158; P = 0.017] and the presence of fibrosis (OR: 4.551; 95 %CI: 1.092 - 18.960; P = 0.037) were independent risk factors for perforation. All cases of perforation were managed with nonsurgical treatment. Younger age and abdominal pain appeared to be related to prolonged hospitalization. CONCLUSION: Tumor size and fibrosis are important factors related to complications during colorectal ESD. Younger age and development of abdominal pain can predict the hospital course in patients with perforation after ESD.

Dependence of particle volume fraction on sound velocity and attenuation of EPDM composites.

The sound velocity and the attenuation coefficient of EPDM (Ethylene-propylene Diene Monomer) composites incorporated with Silicon Carbide particles (SiCp's) of various volume fractions (0-40%) were experimentally and theoretically investigated. For the experiment a through-transmission technique was used. For the theoretical prediction, some mechanical property models such as Reuss model and Coherent Potential Approximation (CPA) model etc. were employed. The experimental results showed that the sound velocity decreased with the increase of the SiCp volume fraction up to 30% and then increased with the 40 vol% specimen. The attenuation coefficient was increased with the increasing SiCp volume fractions. The modified Reuss model with a longitudinal elastic modulus predicted most well the experimental sound velocity and elastic modulus results.

Kondo effect in magnetic tunnel junctions.

Tunneling magnetoresistance was found to be suppressed with decreasing temperature for magnetic tunnel junctions (MTJs) oxidized under high plasma power. A strong temperature dependence of the junction resistance was observed, along with zero-bias anomalies of dynamic resistance at low temperatures. Resistance shows a logarithmic dependence on temperature, and resistance versus temperature exhibits a scaling behavior. Our experimental data can be explained in a consistent way by the Kondo effect in the MTJs with the Kondo temperature TK=20-30 K.

Single-Crystalline Diluted Magnetic Semiconductor GaN:Mn Nanowires.

Single-crystalline diluted magnetic semiconductor GaN:Mn nanowires with controlled Mn concentrations have been successfully synthesized and incorporated into devices. These nanowires exhibit Curie temperatures above room temperature, magnetoresistances near room temperature, and spin-dependent transport. The nanowires are used as building blocks for the fabrication of GaN:Mn/n-SiC based light-emitting diodes.

Feasibility of bone assessment with leaky Lamb waves in bone phantoms and a bovine tibia.

In this study, the effect of cortical thickness variation on the propagation of leaky Lamb waves is investigated by using an axial transmission technique commonly used to characterize long bones. Three Lucite plates with thicknesses of 1, 3, and 5 mm as bone phantoms and one bovine tibia with a cortical thickness of 2 mm were used at various low frequencies. Experimental measurements in bone phantoms show that the peak frequency and amplitude of excited Lamb modes strongly depend on the thickness of the Lucite plate. In the bovine tibia, the S0 and A0 Lamb modes are consistently observed in the frequency-thickness region from 0.2 to 1.0 MHz mm, and can be effectively launched at a frequency of 200 kHz, suggesting 200 kHz to be the optimal signal frequency for in vivo clinical applications. It can be also seen that both modes are affected by the frequency-thickness product, but the effect is greater for the A0 mode. Hence, the A0 Lamb mode seems more sensitive to cortical thickness change due to aging and osteoporosis. This study suggests that the use of leaky Lamb waves is feasible for ultrasonic bone assessment.

Grb2 dominantly associates with dynamin II in human hepatocellular carcinoma HepG2 cells.

The two SH3 domains and one SH2 domain containing adaptor protein Grb2 is an essential element of the Ras signaling pathway in multiple systems. The SH2 domain of Grb2 recognizes and interacts with phosphotyrosine residues on activated tyrosine kinases, whereas the SH3 domains bind to several proline-rich domain-containing proteins such as Sos1. To define the difference in Grb2-associated proteins in hepatocarcinoma cells, we performed coprecipitation analysis using recombinant GST-Grb2 fusion proteins and found that several protein components (p170, p125, p100, and p80) differently associated with GST-Grb2 proteins in human Chang liver and hepatocarcinoma HepG2 cells. Sos1 and p80 proteins dominantly bind to Grb2 fusion proteins in Chang liver, whereas p100 remarkably associate with Grb2 in HepG2 cells. Also GST-Grb2 SH2 proteins exclusively bound to the p46(Shc), p52(Shc), and p66(Shc) are important adaptors of the Ras pathway in HepG2 cells. The p100 protein has been identified as dynamin II. We observed that the N-SH3 and C-SH3 domains of Grb2 fusion proteins coprecipitated with dynamin II besides Sos1. These results suggest that dynamin II may be a functional molecule involved in Grb2-mediated signaling pathway on Ras activation for tumor progression and differentiation of hepatocarcinoma cells.

Negative regulation of YY1 transcription factor on the dynamin I gene promoter.

Dynamin I is highly expressed in brain and plays a critical role in clathrin-mediated endocytosis and synaptic vesicle recycling. To elucidate the molecular mechanism by which expression of dynamin I is tissue-specifically regulated, we previously cloned and characterized the promoter of the mouse dynamin I gene and suggested that there is a negative regulatory element in this promoter region. In the present study, we showed that YY1 binds to this negative regulatory element located at -111 to -107 by using the EMSA and supershift analyses. Cotransfection experiment using an YY1 expression vector revealed that YY1 exerts a repressive role on the dynamin I gene promoter activity. These results demonstrate that transcription factor YY1 negatively regulates dynamin I expression via binding to the negative regulatory element.

The neuron restrictive silencer factor can act as an activator for dynamin I gene promoter activity in neuronal cells.

The neuron restrictive silencer element (NRSE) has been identified in several neuronal genes and confers neuron specificity by silencing transcription in nonneuronal cells. We have previously reported that Sp1 and an NF-kappaB-like element (NE-1) are required for the promoter activity of mouse dynamin I gene. In the present study, we found that the upstream regulatory region of the dynamin I promoter has an NRSE-like sequence and showed that neuron restrictive silencer factor (NRSF) binds to this element in neuronal cells as well as in nonneuronal cells. We also showed that NRSF activates the promoter activity of dynamin I gene in neuronal cells. From the results in this study, we suggest that NRSE might be involved in the neuron restriction of dynamin I expression, and NRSF could act as an activator for promoter activity of dynamin I gene in neuronal cells.

Increased GTP-binding to dynamin II does not stimulate receptor-mediated endocytosis.

Regarding the molecular mechanism of dynamin in receptor-mediated endocytosis, GTPase activity of dynamin has been thought to have a critical role in endocytic vesicle internalization. However, a recent report suggested that GTP-binding to dynamin itself activates the dynamin to recruit molecular machinery necessary for endocytosis. In this study, to investigate the role of GTP binding to dynamin II, we generated two mutant dynamin II constructs: G38V and K44E. G38V, its GTP binding site might be mainly occupied by GTP caused by reduced GTPase activity, and K44E mutant, its GTP binding site might be vacant, caused by its decreased affinity for GTP and GDP. From the analysis of the ratio of GTP vs GDP bound to dynamin, we confirmed these properties. To test the effect of these mutant dynamins on endocytosis, we performed flow cytometry and confocal immunofluorescence analysis and found that these two mutants have inhibitory effect on transferrin-induced endocytosis. Whereas fluorescent transferrin was completely internalized in wild-type (WT) dynamin II expressing cells, no intracellular accumulation of fluorescent transferrin was found in the cells overexpressing K44E and G38V mutant. Interestingly, the amount of GTP bound to K44E was increased when endocytosis was induced than that bound to WT. The present results suggested that the GTPase activity of dynamin II is required for formation of endocytic vesicle and GTP-binding to dynamin II per se is not sufficient for stimulating endocytosis.

Characterization of the mouse dynamin I gene promoter and identification of sequences that direct expression in neuronal cells.

Dynamin I is expressed at high levels in brain and its expression is regulated during the developmental stages of brain. To elucidate the molecular mechanism by which the expression is tissue-specifically regulated, we cloned the 5'-flanking region of the mouse dynamin I gene and determined the nucleotide sequence of 1036 bases upstream from the translation start site. Transient transfection studies with a chloramphenicol acetyltransferase reporter gene in neuroblastoma NS20Y and Lewis lung cells demonstrated that the 5'-flanking region has a cell-type-specific promoter activity. Deletion analyses demonstrated that the minimal promoter activity was detected in the proximal region 195 bp upstream of the translation initiation codon (-90 to +105). The minimal promoter was embedded in a GC-rich region (75% GC content), in which an Sp1-binding motif and a nuclear factor (NF)-kappa B-like element (NE-1) were found, but it lacked TATA and CAAT boxes. Mutational analysis and electrophoretic mobility-shift assay analysis revealed that Sp1 binds to the Sp1 site and that this element is critical for the promoter activity of the dynamin I gene. We found that the NE-1 sequence is required for the expression of the dynamin I gene but NEBP (NE-1-binding protein), which binds to the NE-1 sequence, is not NF-kappa B. We also found that one base in the NE-1 sequence (the underlined G residue in GGGATTCGCGGA) is critical for binding specificity to discriminate between NEBP and NF-kappa B. By UV cross-linking analysis, we found that NEBP is an approx. 104 kDa nuclear protein.

Fractal dimension calculated from two types of region of interest.

OBJECTIVES: To compare the effect of using two region of interest (ROI) types on calculating the fractal dimensions of trabecular bone during simulated osteoporosis. METHODS: Ten 5 mm thick cross-sections from the long bone of a cow were progressively decalcified in 0.1 N Hcl for 5, 10, 20, 30, 60 and 90 min intervals, and radiographed using 0 degrees projection angle in a specially designed device. Two types of ROI (the ruggedness of the boundary and bone profiles) were placed on each digital image. Fractal dimensions and variance in mean pixel intensity were computed from each ROI using the caliper method in ImageFractal (National Institutes of Health, Washington, DC, USA). Correlation analysis quantified the relationship between changes in variance and fractal dimensions of the two types of ROI. RESULTS: A Strong correlation (r=0.90 approximately 0.98, P

Effects of green tea tannin on cisplatin-induced nephropathy in LLC-PK1 cells and rats.

A study was conducted to clarify whether green tea tannin ameliorated cisplatin-induced renal injury in terms of lactate dehydrogenase and malondialdehyde leakage from a renal epithelial cell line, swine-derived LLC-PK1 cells in culture. Green tea tannin was shown to suppress the cytotoxicity of cisplatin, the suppressive effect increasing with the dose of green tea tannin. The effect of cisplatin was then investigated in rats given green tea tannin for 40 days before cisplatin administration and in control rats given no green tea tannin. In control rats, blood, urinary and renal parameters and the activities of antioxidative enzymes in renal tissue deviated from the normal range, indicating dysfunction of the kidneys. In contrast, rats given green tea tannin showed decreased blood levels of urea nitrogen and creatinine, and decreased urinary levels of protein and glucose, reflecting less damage to the kidney. In this group, the activity of catalase in the renal tissue was increased, while the level of malondialdehyde was decreased, suggesting the involvement of radicals in the normalizing of kidney function. Based on the evidence available it appeared that green tea tannin eliminated oxidative stress and was beneficial to renal function.

Atom trap in an axicon mirror.

We have realized a novel atom trap in an axicon (conical hollow) mirror, using a frequency-modulated, single-diode laser. Different spatial distributions of trapped atoms such as a ball and a ring are observed. We show that our numerical simulations are consistent with experimental results. In particular, the ring diameter is found to be approximately the separation between the mirror axis and the magnetic field axis. The axicon trap may be useful as a precooled atom source for cold atomic beams, atom funnels, and atom waveguides.

Single-beam atom trap in a pyramidal and conical hollow mirror.

We present a novel and simple magneto-optical trap in pyramidal and in conical hollow mirrors, using a single beam. A diode laser having modulation sidebands at microwaves is used for cooling, trapping, and repumping of rubidium atoms in a vapor cell. When the laser is circularly polarized and sent into the hollow region, three pairs of counterpropagating beams are automatically produced therein that have the same polarization configuration as that of a conventional six-beam magneto-optical trap. The fluorescence by the trapped atoms and its mirror image are observed simultaneously. This system may be useful for atom-manipulation applications such as gravitational atom traps and atom waveguides.

Cellular factors binding to a novel cis-acting element mediate steroid hormone responsiveness of mouse mammary tumor virus promoter.

Steroid hormone receptors regulate mouse mammary tumor virus (MMTV) gene expression by binding to hormone response DNA elements present in the long terminal repeat. Tissue-specific expression of MMTV is unlikely to be regulated by steroid hormone-receptor complex alone, and mammary cell-specific factors might play a role in the hormone-induced transcriptional activation. In this report we have investigated the function of a novel cis-acting element designated Kil (-204 to -188) which is located adjacent to the distal glucocorticoid response element, in steroid hormone-induced transcription of MMTV. Electrophoretic mobility shift assays indicate that cellular factors bind to the Kil element, and dexamethasone stimulation results in alterations in the binding pattern of proteins in this region. By transient transfection assays using wild type and deletion mutants of the Kil element, we show that this novel cis-acting element is necessary for hormone-induced transcription of MMTV and functions in mammary tumor cells but not in NIH/3T3 cells. Mutagenesis of the Kil sequence suggests that the entire Kil element functioning as one unit is necessary for hormone-induced transcription of MMTV. When placed in the context of heterologous promoters, neither Kil element nor glucocorticoid response element is able to induce significant hormone-induced transcription of MMTV. The presence of both the DNA elements in tandem results in optimal induction of transcription in the presence of steroid hormones. Our results also indicate that the Kil element functions in human breast carcinoma cell lines such as T47D and MCF-7. These results suggest that Kil element in combination with distal glucocorticoid response element functions as a mammary cell-specific enhancer to regulate MMTV transcription.