Validated limited gene predictor for cervical cancer lymph node metastases.

PURPOSE: Recognizing the prognostic significance of lymph node (LN) involvement for cervical cancer, we aimed to identify genes that are differentially expressed in LN+ versus LN- cervical cancer and to potentially create a validated predictive gene signature for LN involvement. MATERIALS AND METHODS: Primary tumor biopsies were collected from 74 cervical cancer patients. RNA was extracted and RNA sequencing was performed. The samples were divided by institution into a training set (n = 57) and a testing set (n = 17). Differentially expressed genes were identified among the training cohort and used to train a Random Forest classifier. RESULTS: 22 genes showed > 1.5 fold difference in expression between the LN+ and LN- groups. Using forward selection 5 genes were identified and, based on the clinical knowledge of these genes and testing of the different combinations, a 2-gene Random Forest model of BIRC3 and CD300LG was developed. The classification accuracy of lymph node (LN) status on the test set was 88.2%, with an Area under the Receiver Operating Characteristic curve (ROC-AUC) of 98.6%. CONCLUSIONS: We identified a 2 gene Random Forest model of BIRC3 and CD300LG that predicted lymph node involvement in a validation cohort. This validated model, following testing in additional cohorts, could be used to create a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) tool that would be useful for helping to identify patients with LN involvement in resource-limited settings.

Rhodobacter sphaeroides Extract Lycogen™ Attenuates Testosterone-Induced Benign Prostate Hyperplasia in Rats.

Benign prostate hyperplasia (BPH) is one of the most common urological problems in mid-aged to elderly men. Risk factors of BPH include family history, obesity, type 2 diabetes, and high oxidative stress. The main medication classes for BPH management are alpha blockers and 5α-reductase inhibitors. However, these conventional medicines cause adverse effects. Lycogen™, extracted from Rhodobacter sphaeroides WL-APD911, is an anti-oxidant and anti-inflammatory compound. In this study, the effect of Lycogen™ was evaluated in rats with testosterone-induced benign prostate hyperplasia (BPH). Testosterone injections and Lycogen™ administration were carried out for 28 days, and body weights were recorded twice per week. The testosterone injection successfully induced a prostate enlargement. BPH-induced rats treated with different doses of Lycogen™ exhibited a significantly decreased prostate index (PI). Moreover, the Lycogen™ administration recovered the histological abnormalities observed in the prostate of BPH rats. In conclusion, these findings support a dose-dependent preventing effect of Lycogen™ on testosterone-induced BPH in rats and suggest that Lycogen™ may be favorable to the prevention and management of benign prostate hyperplasia.

Heteronemin Is a Novel c-Met/STAT3 Inhibitor Against Advanced Prostate Cancer Cells.

BACKGROUND: Prostate cancer is one of the most prevalent cancers in men worldwide. Aberrant activation of c-Met/signal transducer and activator of transcription-3 (STAT3) signaling is involved in prostate carcinogenesis, underscoring the demand for developing c-Met/STAT3-targeting drugs. Thus, we first utilized virtual screening strategy to identify STAT3-inhibiting marine compound, heteronemin, and then validated the STAT3-inhibiting function of heteronemin in prostate cancer cells. METHODS: Human prostate cancer LNCaP, DU145, and PC-3 cell lines were treated with heteronemin for 24 hr, then the cell viability was evaluated by MTT assay. Flow cytometry was performed to analyze the apoptosis in heteronemin-treated cells. Western blot and quantitative real-time PCR were executed to further confirm the c-Met/STAT3 signaling inhibition by heteronemin in DU145 and PC-3 cells. RESULTS: In this study, we employed the virtual screening strategy to identify heteronemin, a spongean sesterterpene, as a potential STAT3 inhibitor from Taiwan marine drugs library. Application of heteronemin potently suppressed the viability and anchorage-independent growth of human prostate cancer cells. Besides, heteronemin induced apoptosis in prostate cancer cells by activation of both intrinsic (caspase-9) and extrinsic (caspase-8) apoptotic pathways. By luciferase assay and expression analysis, it was confirmed that heteronemin inhibited the phosphorylation of c-Met/src/STAT3 signaling axis, STAT3-driven luciferase activities and expression of STAT3-regulated genes including Bcl-xL, Bcl-2, and Cyclin D1. Finally, heteronemin effectively antagonized the hepatocyte growth factor (HGF)-stimulated c-Met/STAT3 activation as well as the proliferation and colonies formation in refractory prostate cancer cells. CONCLUSIONS: These findings suggest that heteronemin may constitute a novel c-Met/STAT3-targeting agent for prostate cancer. Prostate 76:1469-1483, 2016. © 2016 Wiley Periodicals, Inc.

Approaching Defect-free Amorphous Silicon Nitride by Plasma-assisted Atomic Beam Deposition for High Performance Gate Dielectric.

In the past few decades, gate insulators with a high dielectric constant (high-k dielectric) enabling a physically thick but dielectrically thin insulating layer, have been used to replace traditional SiOx insulator and to ensure continuous downscaling of Si-based transistor technology. However, due to the non-silicon derivative natures of the high-k metal oxides, transport properties in these dielectrics are still limited by various structural defects on the hetero-interfaces and inside the dielectrics. Here, we show that another insulating silicon compound, amorphous silicon nitride (a-Si3N4), is a promising candidate of effective electrical insulator for use as a high-k dielectric. We have examined a-Si3N4 deposited using the plasma-assisted atomic beam deposition (PA-ABD) technique in an ultra-high vacuum (UHV) environment and demonstrated the absence of defect-related luminescence; it was also found that the electronic structure across the a-Si3N4/Si heterojunction approaches the intrinsic limit, which exhibits large band gap energy and valence band offset. We demonstrate that charge transport properties in the metal/a-Si3N4/Si (MNS) structures approach defect-free limits with a large breakdown field and a low leakage current. Using PA-ABD, our results suggest a general strategy to markedly improve the performance of gate dielectric using a nearly defect-free insulator.

The Functional Haplotypes of CHRM3 Modulate mRNA Expression and Associate with Bladder Cancer among a Chinese Han Population in Kaohsiung City.

Bladder cancer is one of the major cancer types and both environmental factors and genetic background play important roles in its pathology. Kaohsiung is a high industrialized city in Taiwan, and here we focused on this region to evaluate the genetic effects on bladder cancer. Muscarinic acetylcholine receptor M3 (CHRM3) was reported as a key receptor in different cancer types. CHRM3 is located at 1q42-43 which was reported to associate with bladder cancer. Our study attempted to delineate whether genetic variants of CHRM3 contribute to bladder cancer in Chinese Han population in south Taiwan. Five selected SNPs (rs2165870, rs10802789, rs685550, rs7520974, and rs3738435) were genotyped for 30 bladder cancer patients and 60 control individuals and genetic association studies were performed. Five haplotypes (GTTAT, ATTGT, GCTAC, ACTAC, and ACCAC) were found significantly associated with low CHRM3 mRNA level and contributed to increased susceptibility of bladder cancer in Kaohsiung city after rigid 10000 consecutive permutation tests. To our knowledge, this is the first genetic association study that reveals the genetic contribution of CHRM3 gene in bladder cancer etiology.

Spinodal decomposition of mono- to few-layer graphene on Ni substrates at low temperature.

Mono to few-layer graphene were prepared on pre-annealed polycrystalline nickel substrates by chemical vapor deposition at a relatively low temperature of 800 degrees C using fast cooling rate. It was observed that the reduced solubility of Carbon in Ni at low temperature and an optimum gas mixing ratio (CH4:H2 = 60/80 (sccm)) can be used to synthesize mano-layer graphene that covers about 100 microm2 area. The number of graphene layers strongly depends upon the hydrogen and methane flow rates. An increase in the methane flow is found to increase the growth density of the single-layer graphene. The number of graphene layers was identified from micro-Raman spectra. The thinnest areas containing mono-layer graphene formed at small Ni grains surrounded by large Ni Grains can be explained in terms of Spinodal decomposition. Scanning tunneling microscopy observations of the graphene samples indicate that the graphene structure exhibits no defects, and extremely symmetry hexagon carbon at flat graphene surface is observed.

Percutaneous vertebral augmentation: an elevation in adjacent-level fracture risk in kyphoplasty as compared with vertebroplasty.

BACKGROUND CONTEXT: Osteoporotic vertebral compression fractures (VCFs) are being increasingly treated with minimally invasive bone augmentation techniques such as kyphoplasty and vertebroplasty. Both are reported to be an effective means of pain relief; however, there may be an increased risk of developing subsequent VCFs after such procedures. PURPOSE: The purpose of this study was to compare the effectiveness and complication profile of kyphoplasty and vertebroplasty in a single patient series. STUDY DESIGN/SETTING: A clinical series of 36 patients with VCFs treated by vertebral augmentation procedures was retrospectively analyzed for surgical approach, volume of cement injected, cement extravasation (symptomatic and asymptomatic), the occurrence of subsequent adjacent level fracture, and pain relief. PATIENT SAMPLE: Thirty-six patients with 46 VCFs underwent either kyphoplasty or vertebroplasty after failing conservative therapy. The mean patient age was not significantly different between the kyphoplasty group (70; range, 46-83) and vertebroplasty group (72; range, 38-90) (p=.438). OUTCOME MEASURES: Outcomes were assessed by using self-report measures (a comparative pain rating scale) and physiologic measures (pre- and postoperative radiographs). METHODS: Thirty-six patients with VCFs underwent 46 augmentation procedures (17 patients had 20 fractures treated via kyphoplasty, and 19 patients had 26 fractures treated via vertebroplasty). Seventeen patients in this series underwent kyphoplasty using standard techniques involving bone void creation with balloon tamps, followed by cement injection. Nineteen patients underwent a percutaneous vertebroplasty procedure using a novel cannulated, fenestrated bone tap developed to direct cement anteriorly into the vertebral body to avoid backflow of cement onto neural elements. RESULTS: Pain improvement was seen in >90% of patients in both groups. Mean cement injection per vertebral body was 4.65 mL and 3.78 mL for the kyphoplasty and vertebroplasty groups, respectively (p=.014). Ninety-five percent of the kyphoplasty procedures were performed bilaterally, whereas only 19% of the vertebroplasty procedures required bilateral augmentation (p<.001). There was no cement extravasation resulting in radiculopathy, or myelopathy in either group. Asymptomatic cement extravasation was seen in 5 of 46 (11%) of the total series (3/20 [15%] and 2/26 [7.7%] of kyphoplasty and vertebroplasty, respectively) (p=.696). Within a 3-month period, there were 5 new adjacent level fractures seen in 3 patients who underwent a kyphoplasty procedure (5/20 [25%]) and none in the vertebroplasty group (p<.05). CONCLUSIONS: Vertebroplasty appears to offer a comparable rate of postoperative pain relief as kyphoplasty while using less bone cement more often via a unilateral approach and without the attendant risk of adjacent level fracture.

Segmental polymethylmethacrylate-augmented pedicle screw fixation in patients with bone softening caused by osteoporosis and metastatic tumor involvement: a clinical evaluation.

OBJECTIVE: Instrumentation of the osteoporotic spine may result in bone failure because of pedicle screw loosening and pullout. A clinical evaluation of a novel fenestrated bone tap used in pedicle screw augmentation was performed to determine the performance and safety of this technique. METHODS: Over a 2.5-year period, the clinical and radiographic results of 119 consecutive patients who underwent instrumented arthrodesis were reviewed. Of these patients, 23 had bone softening secondary to osteoporosis and/or metastatic spinal tumor involvement. These patients underwent surgical decompression and spinal instrumentation. RESULTS: Six patients (26%) had metastatic spine disease (squamous cell lung carcinoma, renal cell carcinoma, bladder carcinoma, breast, prostate, and uterine adenocarcinoma); five patients (22%) had a degenerative spondylolisthesis; and 12 patients (52%) had burst fractures, eight as a result of benign causes and four as a result of metastatic disease. Four (17%) patients underwent revision surgery of previous pedicle screw failure resulting from bone softening and pseudarthrosis. A total of 98 levels were fused using 158 polymethylmethacrylate-augmented screws. None of the patients experienced operative death, myocardial infarction, hypoxemia, intraoperative hypotension, radiculopathy, or myelopathy. Asymptomatic anterior cement extravasation was observed in nine patients (39%). There was one asymptomatic polymethylmethacrylate pulmonary embolus and one wound infection. There was no significant relationship between cement extravasation and the quantity used, levels augmented, or location (P > 0.05). There were no construct failures. CONCLUSION: Polymethylmethacrylate-augmented pedicle screw fixation reduces the likelihood of pedicle screw loosening and pullout in patients with osteoporosis requiring instrumented arthrodesis.

A biomechanical cadaveric analysis of polymethylmethacrylate-augmented pedicle screw fixation.

OBJECT: Instrumentation of the osteoporotic spine can be fraught with complications such as hardware failure. A cadaver study was performed to determine the biomechanical performance of polymethylmethacrylate (PMMA)-augmented pedicle screws. METHODS: Three osteoporotic human cadaveric specimens with a mean bone mineral density of 0.70 g/cm2 were used to evaluate the performance of a novel fenestrated bone tap in pedicle screw augmentation. On this device, tap threads serve a dual purpose in preventing backflow of cement toward neural elements while allowing for a custom form for subsequent screw placement. The tap was used to inject a mean volume of 3.7 ml PMMA/pedicle (range 2-8.0 ml PMMA/pedicle) followed by pedicle screw placement between L-5 and T-5, alternating between augmented and nonaugmented instrumentation. Axial pullout testing was then performed. RESULTS: Pedicle screw pullout strength was increased in both primary and salvage procedures by 119% (p = 0.001) and 162% (p = 0.01), respectively, after PMMA augmentation. Additionally, the injected cement volumes were divided into two groups, a low-cement group (< or = 2.8 ml/pedicle) and a high-cement group (> or = 5.5 ml/pedicle). Interestingly, the pullout strength did not significantly change with increased cement usage between the two groups (p > 0.05 for all comparisons). CONCLUSIONS: Polymethylmethacrylate-augmented pedicle screw fixation results in a significant increase in the axial pullout strength of augmented pedicle screws in both primary and revision procedures. This technique may be a valuable adjunct in cases in which bolstering of the screw-bone interface is necessary.

DNA loop repair by Escherichia coli cell extracts.

The nick-directed DNA repair efficiency of a set of M13mp18-derived heteroduplexes containing 8-, 12-, 16-, 22-, 27-, 45-, and 429-nucleotide loops was determined by in vitro assay. Unpaired nucleotides of each heteroduplex reside within overlapping recognition sites for two restriction endonucleases, permitting independent evaluation of repair occurring on either DNA strand. Our results show that a strand break located either 3' or 5' to the loop is sufficient to direct heterology repair to the nicked strand in Escherichia coli extracts. Strand-specific repair by this system requires Mg2+ and the four dNTPs and is equally efficient on insertions and deletions. This activity is distinct from the MutHLS mismatch repair pathway. Strand specificity and repair efficiency are largely independent of the GATC methylation state of the DNA and presence of the products of mismatch repair genes mutH, mutL, and mutS. This study provides evidence for a loop repair pathway in E. coli that is distinct from conventional mismatch repair.

A prostate-derived cDNA that is mapped to human chromosome 19 encodes a novel protein.

The epithelial cells of prostate gland secrete various secretory products that play an important role in the growth and differentiation of prostate gland. These secretory products have also been implicated in neuroendocrine differentiation of benign prostatic hyperplasia and prostate malignancy. We have cloned a prostate-derived cDNA encoding a novel protein with a predicted molecular weight of 78 kDa (P(78)), and precisely mapped the cDNA sequence to chromosome 19. The P(78) gene has a complex genomic structure with 18 exons and 17 introns. The P(78) contains two conserved structural domains with limited similarity to domain D of synapsin I. The P(78) mRNA was expressed in various human cell lines. Western blot analysis using antibody specific for the P(78) revealed the presence of the P(78) protein in the prostate cancer cell lines with much lower level in metastatic prostate cancer cell lines compared to that in a primary prostate cancer cell line.

Cloning a cDNA encoding an alternatively spliced protein of BRCA2-associated factor 35.

Inheritance of mutations in the breast cancer susceptibility gene, BRCA2, predisposes humans to breast and ovarian cancers. Inherited mutations in the BRCA2 gene are also known to cause susceptibility to prostate cancer. BRCA2 protein exists in a large multi-protein complex from which a novel structural DNA binding protein BRCA2-associated factor 35 (BRAF35) has been isolated. We have cloned a novel cDNA encoding an alternatively spliced protein of BRAF35, designated as BRAF25. BRAF25 transcript is present in various human cells. We have precisely mapped the BRAF25 cDNA sequence to the genomic chromosome 19 sequence. Analysis of the predicted sequence of BRAF25 identified a protein of 215 amino acids. BRAF25 contains a truncated high mobility group domain, a kinesin-like coiled-coil domain and multiple Src homology 2 (SH2) motifs. Western blot analysis using antibodies specific for BRAF25 revealed the presence of BRAF25 in human prostate cancer cells.

Immunohistological detection of BRAF25 in human prostate tumor and cancer specimens.

BRAF25 is an alternatively spliced protein of BRAF35 (see associated paper). We have mapped the BRAF25 gene to chromosome sub-band 19p13.3, a region where loss of chromosomal heterozygosity has been reported in about 50% of ovarian cancers. Because of the high incidence of genetic links of prostate cancer to breast and ovarian cancers, we investigated the BRAF25 expression in the prostate specimens. Immunohistochemical analysis using antibodies specific for BRAF25 revealed a strong immunostaining in sections of the benign prostatic hyperplasia (BPH). The staining was concentrated on the nuclei of cells facing the lumen of prostatic glands, even though the sporadic nuclei of cells in stromas were also stained. However, the expression of BRAF25 was dramatically reduced in intermediate prostate cancer and absent in advanced prostate cancer. Preincubation of the antibody with the immunizing peptide abolished immunostaining in BPH specimens. Therefore, the expression of BRAF25 was gradually lost in prostate cancer.