Application of next-generation sequencing to improve cancer management: A review of the clinical effectiveness and cost-effectiveness.

Uptake of next-generation sequencing (NGS) has increased dramatically due to significant cost reductions and broader community acceptance of NGS. To systematically review the evidence on both the clinical effectiveness and the cost-effectiveness of applying NGS to cancer care. A systematic search for full-length original research articles on the clinical effectiveness and cost-effectiveness of NGS in MEDLINE and EMBASE. Articles that focussed on cancer care and involved the application of NGS were included for the review of clinical effectiveness. For the cost-effectiveness review, we only included the articles with economic evaluations of NGS in cancer care. We report the rate of successfully detecting mutations from the clinical studies. The incremental cost-effectiveness ratio and sensitivity analysis outcomes are reported for the cost-effectiveness articles. Fifty-six articles reported that sequencing patient samples using targeted gene panels, and 83% of the successfully sequenced patients harboured at least 1 mutation. Only 6 studies reported on the cost-effectiveness of the application of NGS in cancer care. NGS is an effective tool for identifying mutation in cancer patients. However, more rigorous cost-effectiveness studies of NGS applied to cancer management are needed to determine whether NGS can improve patient outcomes cost-effectively.

Aqueous humor endothelin-1 and total retinal blood flow in patients with non-proliferative diabetic retinopathy.

PurposeThe purpose of this study was to determine the association between aqueous ET-1 levels and total retinal blood flow (TRBF) in patients with non-insulin-dependent type 2 diabetes mellitus (T2DM) and early non-proliferative diabetic retinopathy (NPDR).Patients and methodsA total of 15 age-matched controls and 15 T2DM patients with NPDR were recruited into the study. Aqueous humor (~80-120 µl) was collected before cataract surgery to measure the levels of ET-1 using suspension multiplex array technology. Four weeks post surgery, six images were acquired to assess TRBF using the prototype RTVue Doppler FD-OCT (Optovue, Inc., Fremont, CA, USA) with a double circular scan protocol. At the same visit, forearm blood was collected to determine plasma glycosylated hemoglobin (A1c) levels.ResultsAqueous ET-1 was significantly elevated in the NPDR group compared with the control group (3.5±1.8 vs 2.2±0.8, P=0.02). TRBF was found to be significantly reduced in the NPDR group compared with the control group (34.5±9.1 vs 44.1±4.6 µl/min, P=0.002). TRBF and aqueous ET-1 were not correlated within the NPDR group (r=-0.24, P=0.22). In a multivariate analysis, high A1c was associated with reduced TRBF and aqueous ET-1 levels across control and NPDR groups (P<0.01).ConclusionAqueous ET-1 levels were increased while TRBF was reduced in patients with NPDR compared with the control group. Although not directly associated, the vasoconstrictory effects of ET-1 are consistent with a reduced TRBF observed in early DR. ET-1 dysregulation may contribute to a reduction in retinal blood flow during early DR.

Synthesis and monoamine oxidase inhibitory activities of some 3-(4-fluorophenyl)-5-aryl-n-substituted-4,5-dihydro-(1H)-pyrazole-1-carbothioamide derivatives.

28 new 3-(4-fluorophenyl)-5-aryl-N-substituted-4,5-dihydro-1H-pyrazole-1-carbothioamide derivatives were synthesized and evaluated in vitro for their monoamine oxidase (MAO) A and B inhibitory activity and selectivity. The derivatives substituted by halogen on the fifth position of pyrazole ring, inhibited MAO-A enzyme with a high selectivity index. On the other hand, compounds substituted with 2-naphthyl inhibited MAO-B enzyme with a moderate selectivity index. Docking studies were done to highlight the interactions of the most active derivative with the active site of MAO-A. In addition, in vivo antidepressant and anxiolytic activities of the compounds having selective MAO-A inhibitory effects, were investigated by using Porsolt forced swimming and elevated plus-maze tests respectively. 3-(4-Fluorophenyl)-5-(4-chloro-phenyl)-N-allyl-4,5-dihydro-1H-pyrazole-1-carbothio-amide has antidepressant, 3-(4-fluorophenyl)-5-(4-chlorophenyl)-N-methyl-4,5-dihydro-1H-pyrazole-1-carbothioamide and 3-(4-fluoro-phenyl)-5-(4-bromophenyl)-N-ethyl-4,5-dihydro-1H-pyrazole-1-carbothioamide have anxiolytic activity.

Direct effects of Bmi1 on p53 protein stability inactivates oncoprotein stress responses in embryonal cancer precursor cells at tumor initiation.

Embryonal cancer can arise from postnatally persistent embryonal remnant or rest cells, which are uniquely characterized by the absence of p53 mutations. Perinatal overexpression of the MycN oncoprotein in embryonal cancer precursor cells causes postnatal rests, and later tumor formation through unknown mechanisms. However, overexpression of Myc in adult tissues normally activates apoptosis and/or senescence signals as an organismal defense mechanism against cancer. Here, we show that perinatal neuroblastoma precursor cells exhibited a transiently diminished p53 response to MycN oncoprotein stress and resistance to trophic factor withdrawal, compared with their adult counterpart cells from the TH-MYCN(+/+) transgenic mouse model of neuroblastoma. The adult stem cell maintenance factor and Polycomb group protein, Bmi1 (B-cell-specific Moloney murine leukemia virus integration site), had a critical role at neuroblastoma initiation in the model, by repressing p53 responses in precursor cells. We further show in neuroblastoma tumor cells that Bmi1 could directly bind p53 in a complex with other Polycomb complex proteins, Ring1A or Ring1B, leading to increased p53 ubiquitination and degradation. Repressed p53 signal responses were also seen in precursor cells for other embryonal cancer types, medulloblastoma and acute lymphoblastic leukemia. Collectively, these date indicate a general mechanism for p53 inactivation in some embryonal cell types and consequent susceptibility to MycN oncogenesis at the point of embryonal tumor initiation.

TRIM16 acts as a tumour suppressor by inhibitory effects on cytoplasmic vimentin and nuclear E2F1 in neuroblastoma cells.

The family of tripartite-motif (TRIM) proteins are involved in diverse cellular processes, but are often characterized by critical protein-protein interactions necessary for their function. TRIM16 is induced in different cancer types, when the cancer cell is forced to proceed down a differentiation pathway. We have identified TRIM16 as a DNA-binding protein with histone acetylase activity, which is required for the retinoic acid receptor ß(2) transcriptional response in retinoid-treated cancer cells. In this study, we show that overexpressed TRIM16 reduced neuroblastoma cell growth, enhanced retinoid-induced differentiation and reduced tumourigenicity in vivo. TRIM16 was only expressed in the differentiated ganglion cell component of primary human neuroblastoma tumour tissues. TRIM16 bound directly to cytoplasmic vimentin and nuclear E2F1 in neuroblastoma cells. TRIM16 reduced cell motility and this required downregulation of vimentin. Retinoid treatment and enforced overexpression caused TRIM16 to translocate to the nucleus, and bind to and downregulate nuclear E2F1, required for cell replication. This study, for the first time, demonstrates that TRIM16 acts as a tumour suppressor, affecting neuritic differentiation, cell migration and replication through interactions with cytoplasmic vimentin and nuclear E2F1 in neuroblastoma cells.

Pt surface modification of SnO2 nanorod arrays for CO and H2 sensors.

Uniform SnO(2) nanorod arrays were deposited on a 4 inch SiO(2)/Si wafer by plasma-enhanced chemical vapor deposition (PEVCD) at low deposition temperature of around 300 degrees C. The SnO(2) nanorods were connected at the roots, thus the nanorod sensors could be fabricated by a feasible way compatible with microelectronic processes. The surface of the sensors was modified by Pt nanoparticles deposited by dip coating and sputtering, respectively. The sensing properties of the Pt-modified SnO(2) nanorod sensors to CO and H(2) gases were comparatively studied. After surface modification of Pt, the sensing response to CO and H(2) gases increased dramatically. The 2 nm Pt-modified SnO(2) nanorod sensors by sputtering showed the best sensing performance. By increasing Pt thickness from 2 nm up to 20 nm, the optimal working temperature decreased by 30 degrees C while the sensing response also decreased by about 4 times. Comparing these two Pt modification approaches by dip coating and sputtering, both could achieve comparable promotion effect if the Pt thickness can be controlled around its optimal value. The deposition technique of SnO(2) nanorod arrays by PECVD has good potential for scale-up and the fabrication process of nanorod sensors possesses simplicity and good compatibility with contemporary microelectronics-based technology.

High sensitivity SnO2 single-nanorod sensors for the detection of H2 gas at low temperature.

Uniform SnO(2) nanorods were grown by inductively coupled plasma-enhanced chemical vapor deposition without catalysts and additional heating. The SnO(2) nanorods were aligned on a pair of Au/Ti electrodes by the dielectrophoresis method. SnO(2) single-nanorod gas sensors were fabricated by connecting individual SnO(2) nanorods to a pair of Au/Ti electrodes with Pt stripes deposited by a focused ion beam. The sensing properties of the SnO(2) single-nanorod sensor were studied. The SnO(2) single-nanorod sensor could detect 100 ppm H(2) at room temperature with repeated response and showed a large change of resistance, fast response time and good reversibility at an elevated operating temperature of 200 degrees C. The optimal sensing performance of the sensor is achieved at the operating temperature of around 250 degrees C.

Measurement of total blood flow in the normal human retina using Doppler Fourier-domain optical coherence tomography.

AIM: To measure total retinal blood flow in normal human eyes using Doppler Fourier-domain optical coherence tomography (FD-OCT). METHODS: 10 normal people aged 35 to 69 years were measured for the right eye using Doppler FD-OCT. Double circular scans around the optic nerve heads were used. Four pairs of circular scans that transected all retinal branch vessels were completed in 2 s. Total retinal blood flow was obtained by summing the flows in the branch veins. Measurements from the eight scans were averaged. Veins with diameters >33 microm were taken into account. RESULTS: Total retinal blood flow could be measured in eight of 10 subjects: mean (SD) = 45.6 (3.8) microl/min (range 40.8 to 52.9 microl/min). The coefficient of variation for repeated measurements was 10.5%. Measured vein diameters ranged from 33.3 to 155.4 mum. The averaged flow speed was 19.3 (2.9) mm/s, which did not correlate with vessel diameter. There was no significant difference between flows in the superior and inferior retinal hemispheres. CONCLUSIONS: Double circular scanning using Doppler FD-OCT is a rapid and reproducible method to measure total retinal blood flow. These flow values are within the range previously established by laser Doppler flowmetry.

Regulation and activation of ezrin protein in endometriosis.

BACKGROUND: Ezrin protein and its activated form phospho-ezrin play a role in cell morphology, motility and adhesiveness. In this study, we hypothesized that these proteins play a role in the pathogenesis of endometriosis by promoting adhesion and invasion of endometrial stromal cells (ESCs) in ectopic sites. METHODS: We compared the expression of ezrin and phospho-ezrin in normal endometrium from women without endometriosis with their expression in eutopic and ectopic endometrial tissues from women with endometriosis, using immunohistochemistry and western blot analysis. Paired eutopic and ectopic endometrial tissue samples from women with endometriosis (n = 13) and normal endometrium from women without endometriosis (n = 12) were collected. Invasive potential of ESCs from each of these samples was compared using Matrigel membrane invasion assay. RESULTS: Eutopic and ectopic endometrial tissues from women with endometriosis have higher ezrin and phospho-ezrin levels as confirmed by immunohistochemistry and western blot analysis (P < 0.05). The Matrigel membrane invasion assay revealed that ectopic ESCs have more invasive characteristics, more protrusions and higher ezrin staining than normal ESCs (P < 0.05). CONCLUSIONS: Ezrin can be a potential marker for endometrial cell invasion and may play a role in the pathogenesis of endometriosis.

Axial superresolution of two-photon microfabrication.

An axial superresolution diffraction theory is developed in a two-photon microfabrication system. This method can improve the axial superresolution of the two-photon microfabrication system. A theoretical analysis of the photosensitive resin is discussed based on the exciting power and the concentration of free radical theory. Simulated results of the two-photon microfabrication verify the method and show that it can provide insight into the microfabrication system.

Charge trapping phenomena of tetraethylorthosilicate thin film containing Si nanocrystals synthesized by solid-state reaction.

In this work, we report on the fabrication of tetraethylorthosilicate (TEOS) thin dielectric film containing silicon nanocrystals (Si nc), synthesized by solid-state reaction, in a capacitor structure. A metal-insulator-semi-conductor (MIS) capacitor, with 28 nm thick Si nc in a TEOS thin film, has been fabricated. For this MIS, both electron and hole trapping in the Si nc are possible, depending on the polarity of the bias voltage. A V(FB) shift greater than 1 V can be experienced by a bias voltage of 16 V applied to the metal electrode for 1 s. Though there is no top control oxide, the discharge time for 10% of charges can be up to 4480 s when it is biased at 16 V for 1 s. It is further demonstrated that charging and discharging mechanisms are due to the Si nc rather than the TEOS oxide defects. This form of Si nc in a TEOS thin film capacitor provides the possibility of memory applications at low cost.

Compartmentalized expression of kallikrein 4 (KLK4/hK4) isoforms in prostate cancer: nuclear, cytoplasmic and secreted forms.

The prostate-specific antigen-related serine protease gene, kallikrein 4 (KLK4), is expressed in the prostate and, more importantly, overexpressed in prostate cancer. Several KLK4 mRNA splice variants have been reported, but it is still not clear which of these is most relevant to prostate cancer. Here we report that, in addition to the full-length KLK4 (KLK4-254) transcript, the exon 1 deleted KLK4 transcripts, in particular, the 5'-truncated KLK4-205 transcript, is expressed in prostate cancer. Using V5/His6 and green fluorescent protein (GFP) carboxy terminal tagged expression constructs and immunocytochemical approaches, we found that hK4-254 is cytoplasmically localized, while the N-terminal truncated hK4-205 is in the nucleus of transfected PC-3 prostate cancer cells. At the protein level, using anti-hK4 peptide antibodies specific to different regions of hK4-254 (N-terminal and C-terminal), we also demonstrated that endogenous hK4-254 (detected with the N-terminal antibody) is more intensely stained in malignant cells than in benign prostate cells, and is secreted into seminal fluid. In contrast, for the endogenous nuclear-localized N-terminal truncated hK4-205 form, there was less difference in staining intensity between benign and cancer glands. Thus, KLK4-254/hK4-254 may have utility as an immunohistochemical marker for prostate cancer. Our studies also indicate that the expression levels of the truncated KLK4 transcripts, but not KLK4-254, are regulated by androgens in LNCaP cells. Thus, these data demonstrate that there are two major isoforms of hK4 (KLK4-254/hK4-254 and KLK4-205/hK4-205) expressed in prostate cancer with different regulatory and expression profiles that imply both secreted and novel nuclear roles.

Is varicocele a prognostic factor for determining sperm retrieval rate before testicular sperm extraction?

66 nonobstructive azoospermic mean with normal genetic analysis composed of 32 (48%) patients with and 34 (52%) patients without varicocele were evaluated for the rate of sperm extraction five months after the varicocelectomy. Sperm retrieval was successful in 22 of 32 patients (68%) who had been operated because of varicocele and in 13 of 34 patients (38.2%) who had no varicocele (OR = 3.55) (CI: 1.15-11.27) (p = 0.025). Overall, sperm extraction was successful in 35 of 66 patients (53%). Repair of varicocele in non-obstructive azoospermic patients may return spermatogenesis to normal in spermatogenic focuses. So, the present of varicocele and its treatment might be considered as a prognostic factor for sperm retrieval in these patients.

Proliferating cell nuclear antigen immunoreactivity in the ovarian surface epithelium of mice of varying ages and total lifetime ovulation number following ovulation.

Everytime an oocyte is released at ovulation, the ovarian surface epithelium (OSE) is ruptured and must be restored by epithelial cell proliferation. Ovulation site closure was studied in mice of various ages along with total lifetime ovulation number to investigate the known association of these factors with the risk of epithelial ovarian cancer. Ovaries from Swiss Webster mice were collected at various time points postovulation from 3-mo virgin animals (estimated median total lifetime ovulation number, 92; n = 40 mice), 8-mo virgin animals subject to incessant ovulation (estimated median total lifetime ovulation number, 652; n = 15 mice), and 12-mo breeders (estimated median total lifetime ovulation number, 208; n = 35 mice). Diameters of ovulation sites were estimated by scanning electron microscopy. No differences were found in the rate of ovulation site closure between the groups. Sections of ovaries were analyzed using immunohistochemistry for proliferating cell nuclear antigen (PCNA). The highest density of immunoreactive cells was observed in all animal groups in the cuboidal cells around the rupture site the day after ovulation. Despite the similarity in ovulation site closure rates between groups, the total number of OSE cells that were positive for PCNA in both the 8- and 12-mo animals was significantly reduced, so the number of stained cells appeared to be insufficient to cover the ovulation site. These data suggest that other mechanisms, such as proliferation of the extraovarian mesothelium, may play a role in the re-epithelialization of the ovary.

Report of a girl with Klippel-feil syndrome and Poland anomaly.

Report of a girl with Klippel-feil syndrome and Poland anomaly: Klippel-Feil syndrome, consisting of the triad of a short neck, low posterior hairline, and limitation of neck movement, is a congenital anomaly characterized by the fusion of cervical vertebrae, Poland anomaly consists of unilateral aplasia of the chest wall muscles and ipsilateral anomalies of upper extremity. We report a 7-year-old girl with typical findings of Klippel-Feil syndrome and Poland anomaly. To the best of our knowledge a case of Klippel-Feil syndrome and Poland anomaly has not been described before, although a combination of Poland, Klippel-Feil and Moebius anomalies has been reported in the literature.

Placenta previa percreta with urinary bladder and ureter invasion.

A 26-year-old woman, with one previous cesarean delivery and two uterine curettage due to incomplete abortion, was admitted to the labor ward with the diagnosis of partial placenta previa at 35 weeks of gestation. Repeat cesarean section was performed due to profuse vaginal bleeding. Placenta previa percreta invading the bladder trigone was confirmed with cystotomy. As bilateral hypogastric artery ligation and supracervical hysterectomy performed were not successful in stopping the profuse bleeding, the abdomen was packed with laparotomy pads. Dilatation of the left ureter was noticed on the second postoperative day. Relaparotomy was performed to remove the pads, and placental invasion of the distal left ureter was noticed. Ureteroneocystostomy was performed. The postoperative course was uneventful, and the double-J-catheter was removed two months later.

Mutations in a novel factor, glomulin, are responsible for glomuvenous malformations ("glomangiomas").

Glomuvenous malformations (GVMs) are cutaneous venous lesions characterized by the presence of smooth-muscle--like glomus cells in the media surrounding distended vascular lumens. We have shown that heritable GVMs link to a 4--6-cM region in chromosome 1p21-22. We also identified linkage disequilibrium that allowed a narrowing of this VMGLOM locus to 1.48 Mb. Herein, we report the identification of the mutated gene, glomulin, localized on the basis of the YAC and PAC maps. An incomplete cDNA sequence for glomulin had previously been designated "FAP48," for "FKBP-associated protein of 48 kD." The complete cDNA for glomulin contains an open reading frame of 1,785 nt encoding a predicted protein of 68 kD. The gene consists of 19 exons in which we identified 14 different germline mutations in patients with GVM. In addition, we found a somatic "second hit" mutation in affected tissue of a patient with an inherited genomic deletion. Since all but one of the mutations result in premature stop codons, and since the localized nature of the lesions could be explained by Knudson's two-hit model, GVMs are likely caused by complete loss of function of glomulin. The abnormal phenotype of vascular smooth-muscle cells (VSMCs) in GVMs suggests that glomulin plays an important role in differentiation of these cells--and, thereby, in vascular morphogenesis--especially in cutaneous veins.

Ureterocystoplasty in bilaterally functional kidneys.

OBJECTIVE: Traditional augmentation cystoplasty using gastrointestinal segments is known to be associated with metabolic abnormalities and alterations in the bladder causing potential carcinogenesis. In this respect alternative techniques have been searched preferably lined by urothelium. We performed ureterocystoplasty in 7 patients with a diagnosis of neurogenic bladder and investigated the clinical and functional aspects. PATIENTS AND METHODS: Between 1995 and 1999, ureterocystoplasty was performed using both ureters in 4 male and 3 female children with bilaterally functional kidneys. Patients' ages varied between 1 and 7 (mean 4.7) years. Before the operation all the children were incontinent, had a small-capacity noncompliant bladder, and high-grade (IV-V, International Classification System) reflux on voiding cystouretrography (VCU). Technetium-99m DTPA renal scintigraphy was also performed in all children to evaluate renal function before and after the operation. RESULTS: Before the operation the mean end-filling intravesical pressure was 45.6 (35-60) cm H(2)O which decreased to 18.9 cm H(2)O 3 months postoperatively. The mean bladder capacity 3 months after ureterocystoplasty was found to be 279.3 (250-330) ml. All the children were continent and VCU showed the absence of reflux. There was mild to moderate improvement in renal function after surgery in both kidneys on technetium-99m DTPA renal scintigraphy. Three (43%) patients could void spontaneously with abdominal straining, whereas 4 (57%) children could empty their bladders by clean intermittent catheterization. A double-J stent was inserted in 1 (14%) patient because of a rise in serum creatinine after the removal of the ureteral catheter. Patients were followed for a mean period of 30 (8-50) months and all the children remained continent. The bladder capacity and end-filling pressure measurements were also stable. CONCLUSION: Ureterocystoplasty was found to be an effective method for bladder augmentation in bilaterally functional kidneys with an acceptable complication rate