BACKGROUND

Significant phenotypical variability is observed in autosomal dominant polycystic kidney disease (ADPKD). Dysregulation of vascular endothelial growth factor (VEGF) expression in the kidney has been demonstrated in a wide range of renal diseases. The aim of the present study was to assess the influence of the -2578 C/A and the -1154 G/A polymorphisms in the regulatory region of the VEGF gene upon the progression of ADPKD toward end-stage renal disease (ESRD).

METHODS

The study was performed on 283 ADPKD patients (145 males, 138 females, mean age 51.7 +/- 10.3 years) who had reached ESRD. Patients were divided into three groups: (1) ESRD development later than in 63 years (slow progressors, n = 47), (2) ESRD development before 45 years (rapid progressors, n = 69), and (3) ESRD development between 45 and 63 years (intermediate progressors, n = 167). Genetically unrelated healthy Czech individuals were analyzed as a control group (n = 311, 153 males, 158 females, mean age 44.6 +/- 9.2 years). DNA samples were genotyped for the -2578 C/A and for the -1154 G/A polymorphisms of the VEGF gene promoter. The serum levels of VEGF were established in 111 healthy Czech individuals from the control group.

RESULTS

The VEGF -2578 C/A and -1154 G/A genotype distribution showed no differences among the groups of slow, rapid and intermediate progressors. The age of ESRD with regard to different genotypes was not significantly different in all ADPKD patients. However, the AA genotype of the -2578 C/A polymorphism was associated with a significantly higher age of ESRD than other genotypes in rapid progressors (42.7 vs. 40.5 years, p = 0.01). The CG haplotype was found significantly more frequent in ADPKD rapid progressors than in slow progressors (p = 0.047). Serum levels of VEGF did not significantly differ in the control group, according to different genotypes of both polymorphisms.

CONCLUSIONS

To conclude, AA genotype of the -2578 C/A polymorphism was related to better prognosis of the disease in a limited group of ADPKD patients. Classical genetic recessive and dominant model did not find significant influence of separate VEGF polymorphisms on the progression of ADPKD. Accordingly, CG haplotype was associated with earlier onset of ESRD in ADPKD patients.

Intimal hyperplasia (IH) is a common cause of vasculopathy due to direct <em>endothelial</em> damage (such as post-coronary re<em>vascular</em>ization) or indirect injury (such as chronic kidney disease, or CKD). Although the attenuation of coronary re<em>vascular</em>ization-induced IH (direct-<em>vascular</em>-injury-induced IH) by cilostazol, a phosphodiesterase III inhibitor, has been demonstrated, our understanding of the effect on CKD-induced IH (indirect-<em>vascular</em>-injury-induced IH) is limited. Herein, we tested if cilostazol attenuated CKD-induced IH in a mouse model of ischemic-reperfusion injury with unilateral nephrectomy (Chr I/R), a normotensive non-proteinuria CKD model. Cilostazol (50 mg/kg/day) or placebo was orally administered once daily from 1-week post-nephrectomy. At 20 weeks, cilostazol significantly attenuated aortic IH as demonstrated by a 34% reduction in the total intima area with 50% and 47% decreases in the ratios of tunica intima area/tunica media area and tunica intima area/(tunica intima + tunica media area), respectively. The diameters of aorta and renal function were unchanged by cilostazol. Interestingly, cilostazol decreased miR-221, but enhanced miR-143 and miR-<em>145</em> in either in vitro or aortic tissue, as well as attenuated several pro-inflammatory mediators, including asymmetrical dimethylarginine, high-sensitivity C-reactive protein, <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> in aorta and serum pro-inflammatory cytokines (IL-6 and TNF-α). We demonstrated a proof of concept of the effectiveness of cilostazol in attenuating IH in a Chr I/R mouse model, a CKD model with predominantly indirect-<em>vascular</em>-injury-induced IH. These considerations warrant further investigation to develop a new primary prevention strategy for CKD-related IH.

BACKGROUND

Vascular endothelial growth factor (VEGF) is a key regulator of angiogenesis and is implicated in the development of diabetic microvascular and macrovascular disease.

METHODS

The expression of total VEGF, VEGF splice variants (VEGF(121), VEGF(145), VEGF(148), VEGF(165), VEGF(183) and VEGF(189)), VEGFR-1 and VEGFR-2, was investigated in biopsies from the right atrium and left internal mammary artery (LIMA) of 32 non-diabetic and 20 diabetic patients undergoing coronary artery bypass grafting.

RESULTS

Diabetes was independently negatively correlated to total VEGF mRNA expression in atrium. Total VEGF, VEGF(121) and VEGF(165) mRNA levels were upregulated in the LIMA of diabetics vs. non-diabetics. The expression of VEGF receptors in atrium and LIMA was similar between these groups. VEGF(121) and VEGF(165) were the major variants expressed, followed by VEGF(189) and VEGF(183), while VEGF(148) and VEGF(145) were detected in small amounts. The expression profile of VEGF splice variants displayed significant heterogeneity between the examined tissues.

CONCLUSIONS

This is the first study to quantify VEGF splice variants expression in cardiac and vascular tissue. Our results could help elucidate the role of VEGF splice variants in diabetic complications.

<AbstractText>To assess whether publication of Comparison of Age-related macular degeneration Treatment Trial (CATT) results and introduction of aflibercept to the marketplace affected intravitreal bevacizumab and ranibizumab utilization.</AbstractText><AbstractText>Retrospective analysis of treatment patterns.</AbstractText><AbstractText>We calculated weekly bevacizumab and ranibizumab utilization during 3 timeframes: (1) before CATT publication, (2) between CATT publication (April 28, 2011) and assignment of a unique aflibercept billing code (January 1, 2013), and (3) afterward for 164,188 Medicare beneficiaries with neo<em>vascular</em> macular degeneration receiving ≥1 anti-<em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> injection(s) from January 1, 2008 to December 31, 2014. We identified ophthalmologists who predominantly (≥80%) administered bevacizumab or ranibizumab and evaluated changes in preferences over the 3 periods. We replicated analyses on 881,381 commercially insured beneficiaries.</AbstractText><AbstractText>Among 317 ophthalmologists administering predominantly ranibizumab to Medicare beneficiaries pre-CATT, 221 (69.7%) reduced ranibizumab use post-CATT, whereas 96 (30.3%) continued using ranibizumab ≥80% of the time. Findings were reversed among 1041 ophthalmologists who predominantly administered bevacizumab pre-CATT-777 (74.6%) continued bevacizumab-predominant use while 264 (25.4%) reduced bevacizumab use post-CATT. Among the <em>145</em> ophthalmologists who predominantly administered ranibizumab before aflibercept's availability, 77 (53.1%) reduced ranibizumab utilization and 68 (46.9%) continued using ranibizumab ≥80% of the time after aflibercept became available. Corresponding numbers among the 909 ophthalmologists who predominantly administered bevacizumab pre-aflibercept were 381 (41.9%) reducing and 528 (58.1%) continuing bevacizumab-predominant use. Similar results were observed for commercially insured patients.</AbstractText><AbstractText>Many ophthalmologists who favored ranibizumab switched to bevacizumab after CATT publication, while most who favored bevacizumab before CATT publication continued favoring it afterward. Aflibercept's introduction had little impact on preferences for ranibizumab or bevacizumab.</AbstractText>

Weight loss and muscle wasting are of critical importance to cancer patients because of their negative effects on survival, functional status, and tolerability of chemotherapy. Because previous data suggest <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> receptor inhibitors disrupt skeletal muscle pathways, such as PI3K and AKT, the current study explored weight loss and muscle wasting in colorectal cancer patients treated with bevacizumab. Patients were assessed for serial weight and radiographic changes in skeletal muscle at baseline and again within 3 months of starting cancer therapy. Computed tomography scans were used to assess muscle. Fifty-seven patients are the focus on this report. These patients manifested a decline in mean weight from 85 to 83 kilograms (P = 0.002). Mean skeletal muscle area at the L3 vertebral level dropped from 148 cm(2) to <em>145</em> cm(2) (P = 0.02). This drop in weight and skeletal muscle occurred independently of cancer progression. No statistically significant differences in survival were observed based on loss of weight or skeletal muscle. Colorectal cancer patients prescribed bevacizumab appear to lose weight and muscle over a few months even in the absence of cancer progression.

The prognostic value of the expression of <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> (VEGF), VEGF receptor 2 (VEGFR2), platelet-derived <em>growth</em> <em>factor</em> (PDGF)-β, and PDGF receptor (PDGFR)-β in papillary renal cell carcinoma (pRCC) is unknown. A total of <em>145</em> patients, who were confirmed to have pRCC, were analyzed. Expression levels of molecular markers were assessed via immunohistochemistry. The median follow-up period for all patients was 52.0 (interquartile range, 34.5-90.5) months. Among the cohort of <em>145</em> patients, high VEGF expression was observed in 100 (69.0%) patients, whereas high expression of VEGFR2, PDGF-β, and PDGFR-β was observed in 64 (44.1%), 42 (29.0%), and 30 (20.7%) patients, respectively. Only patients with high VEGFR2 expression exhibited improved 10-year recurrence-free survival (85.3% versus 58.1%; P=.005) and cancer-specific survival (86.4% versus 70.1%; P=.014) rates compared with individuals who exhibited low expression. Multivariate analysis revealed that high VEGFR2 expression was an independent prognostic <em>factor</em> for recurrence (hazard ratio, 0.326; P=.006) and cancer-specific mortality (hazard ratio, 0.334; P=.046). During follow-up, 17 patients received targeted drug therapy. Patients with high VEGFR2 expression showed a better initial response (partial response, 40%; stable disease, 20%; progressive disease, 40%) than patients with low expression did (partial response, 0%; stable disease, 58.3%; progressive disease, 41.7%; P=.052). pRCC with high VEGFR2 expression seems to be associated with a better initial response to targeted drug therapy and a better prognostic outcome.

Increased <em>vascular</em> permeability and <em>endothelial</em> cell <em>growth</em> are important in the pathogenesis of otitis media with effusion (OME) and the <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> (VEGF) is known to play an important role in the increased <em>vascular</em> permeability and angiogenesis. To date, at least five isoforms of the VEGF family have been identified as VEGF transcripts, encoding polypeptides of 206, 189, 165, <em>145</em> and 121, but their physiological roles are unclear. The purpose of this study was to investigate the expression of VEGF, in both endotoxin-induced OME of the rat and human otitis media. We instilled endotoxin and saline as a control into the middle ear cavity of the rat. Middle ear mucosa were taken at 0 h, 1 h, 3 h, 6 h, 12 h, 1 day, 3 days, 7 days and 14 days and the expression of VEGF mRNA and VEGF protein was evaluated using semi-quantitative RT-PCR and immunohistochemistry. Expression of VEGF164 mRNA and VEGF120 mRNA was first identified 1 h after endotoxin instillation and was dramatically increased over the period 6 h-1 day and then progressively decreased by day 7. The level of expression of VEGF120 mRNA was slightly higher than that of VEGF164 mRNA and that of VEGF164 mRNA was much higher than that of VEGF188 mRNA. Immunostaining revealed expression of VEGF during 6 h to day 3 and its expression was localized to ciliated cells and some inflammatory cells. We also performed RT-PCRs of cDNA from middle ear fluids of 8 human OME patients and middle ear mucosa of 4 chronic otitis media patients for the identification of VEGF mRNA expression. VEGF121 mRNA was highly expressed in all samples compared with VEGF165 mRNA. These results suggest that VEGF may be primarily responsible for increased <em>vascular</em> permeability and <em>endothelial</em> cell <em>growth</em> in OME and that VEGF seems to play a significant role in the pathogenesis of OME.

OBJECTIVE

To investigate the relationship between vascular endothelial growth factor (VEGF) microvascular density (MVD), lymph node metastasis and prognosis of breast carcinoma (BC).

METHODS

VEGF protein expression and MVD in 92 cases of BC and VEGF mRNA expression in part of the cases were studied by immunohistochemistry methods and reverse-transcription polymerase chain reaction (RT-PCR) technique.

RESULTS

VEGF mRNA expression in BC tumor tissues were higher than those in adjacent normal tissues. Increment of both VEGF(121) and VEGF(165) showed significant difference (P < 0.05 approximately 0.01). In addition, VEGF(145) expression was also observed in BC. There was a close positive correlation between VEGF and MVD (r = 0.702, P < 0.01). VEGF protein expression and MVD correlated significantly with lymph node metastasis and tumor relapse (P < 0.05 approximately 0.01). The time period of relapse-free-survival (RFS) in the patient group with high VEGF expression and MVD was significantly lower than RFS in the group with low VEGF expression and MVD (P < 0.01).

CONCLUSIONS

VEGF is highly related to angiogenesis of BC. The increase of VEGF and MVD may promote lymph node metastasis and relapse of BC. VEGF and MVD may have prognostic value in RFS of BC patients.

Peritoneal dialysis (PD) solution contains high concentrations of glucose and glucose degradation products (GDPs). One of several GDPs--3,4-dideoxyglucosone-3-ene (3,4-DGE)--was recently identified as the most reactive and toxic GDP in PD fluids. In vitro, 3,4-DGE has been shown to induce mesothelial cell damage; however, its role in peritoneal fibrosis in vivo remains unclear. In the present study, we intraperitoneally administered chlorhexidine gluconate (CG) for mild peritoneal injury, and we then injected 3,4-DGE [38 μmol/L (low concentration) or <em>145</em> μmol/L (high concentration)] 5 times weekly for 4 weeks. Significant thickening of the parietal peritoneal membrane was observed only when treatment with low or high concentrations of 3,4-DGE occurred after CG administration, but not when either CG or 3,4-DGE alone was given. The combination of CG and 3,4-DGE also caused upregulation of messenger RNA expression of transforming <em>growth</em> <em>factor</em> β1, connective tissue <em>growth</em> <em>factor</em>, fibronectin, collagen type 1 α1 chain, alpha smooth muscle actin (α-SMA), <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> 164, NADPH oxidase 1 and 4, p22phox, p47phox, and gp91phox in peritoneal tissue. Treatment with CG alone was sufficient to cause significant F4/80-positive macrophage infiltration, appearance of α-SMA-positive cells, and vessel formation in the submesothelial layer. Addition of 3,4-DGE markedly enhanced those changes and induced apoptosis, mainly in leukocytes. The concentration of 3,4-DGE in the abdominal cavity declined more rapidly in CG-treated mice than in PBS-treated mice. Peritoneal membrane permeability determined by peritoneal equilibration test showed high transport conditions in peritoneum treated with both CG and 3,4-DGE. These results indicate that, when mild peritoneal damage is already present, 3,4-DGE causes peritoneal thickening and fibrosis, resulting in deterioration of peritoneal membrane function.

OBJECTIVE

To investigate the effects of mifepristone on cell proliferation of human androgen-independent prostate carcinoma cell lines DU-<em>145</em>, PC-3 in vitro and the possible mechanisms involved.

METHODS

The A values of the prostate cancer cells DU-<em>145</em> and PC-3 in each group with various concentrations (1, 10, 50, 100 micromol/L) of mifepristone at various time intervals (24-120 h) were detected with the colorimetric 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl tetrazolium bromide assay. The apoptosis rates of the DU-<em>145</em> and PC-3 cells treated with 10 micromol/L of mifepristone for 24 h and 48 h were assessed by flow cytometry analysis technique. Immunohistochemical technique was used to determine the expression of bax, bcl-2 and vascular endothelial growth factor (VEGF) proteins after treatment with 10 micromol/L of mifepristone.

RESULTS

The A values of the cancer cells treated with 1 micromol/L of mifepristone were similar to that of controls, while those of the cells treated with 10 micromol/L, 50 micromol/L and 100 micromol/L of mifepristone were significantly different from that of controls (P < 0.01). Mifepristone markedly inhibited cell proliferation of prostate cancer cells DU-<em>145</em> and PC-3 on a dose- and time-depending manner. The apoptosis rates of 10 micromol/L mifepristone for DU-<em>145</em> cell line at 24 h, 48 h were respectively 15.3%, 30.4% with flow cytometry method and then PC-3 cell line were respectively 22.2%, 32.0%. Immunohistochemical technique showed the expression of bcl-2 and VEGF in the DU-<em>145</em> and PC-3 cells treated with 10 micromol/L of mifepristone were significantly decreased, and the expression of bax was increased.

CONCLUSIONS

Mifepristone can induce apoptosis of androgen-independent prostate cancer cell lines DU-<em>145</em> and PC-3 in vitro. The apoptosis effect is time-and-dose dependent. Mifepristone could initiate a cell death command via apoptotic pathways decreasing the expression of VEGF protein, downregulating the expression of bcl-2 protein and increasing the expression of bax protein.

BACKGROUND

Vascular endothelial growth factor (VEGF) tyrosine kinase inhibitors (TKIs) are a mainstay of treatment for metastatic renal-cell carcinoma. Stool microbiome composition is predictive of response to immunotherapy and cytotoxic chemotherapy. We sought to investigate whether antibiotics targeting Bacteroides species affect progression-free survival (PFS) while receiving first-line VEGF-TKI therapy.

METHODS

Using a retrospective cohort of intermediate- and poor-risk metastatic renal-cell carcinoma patients from the University of Utah, we categorized patients receiving first-line VEGF-TKIs by receipt of antibiotics (Bacteroides spp., non-Bacteroides spp., or none) and assessed PFS by Kaplan-Meier and Cox proportional hazard models.

RESULTS

Of 145 patients, 17 received antibiotics with Bacteroides spp. coverage and 32 patients received antibiotics without Bacteroides spp. coverage. When compared to patients not receiving antibiotics, improved PFS was seen with each additional day antibiotics were prescribed with Bacteroides spp. coverage (hazard ratio = 0.92; 95% confidence interval, 0.83-0.99; P = .04).

CONCLUSIONS

Targeting stool Bacteroides spp. with antibiotics improves PFS in patients receiving first-line VEGF-TKIs in a duration-dependent manner.

<strong class="sub-title"> Background: </strong> Uveal melanoma (UM) is the most common primary intraocular malignancy in adults. It has been demonstrated that microRNA-<em>145</em> (miR-<em>145</em>) is correlated with the progression of various cancers by regulating the expression of multiple target genes, especially a number of genes that regulate angiogenesis and proliferation. However, the underlying mechanisms of miR-<em>145</em> in tumor angiogenesis of UM are still not well illustrated. Thus, we aimed to explore the potential target genes or pathways regulated by miR-<em>145</em> in UM and the effect of miR-<em>145</em> on invasion and angiogenesis.

<strong class="sub-title"> Methods: </strong> Totally, 24 choroid samples were collected in our study, including 12 UM samples and 12 normal uveal tissues. The expression of neuroblastoma RAS viral oncogene homolog (N-RAS), phosphorylated protein kinase B (p-AKT), and vascular endothelial growth factor (VEGF) in UM tissues and normal uveal tissues was analyzed using Western blotting analysis. Lentivirus expression system was used to construct MUM-2B and OCM-1 cell lines with stable overexpression of miR-<em>145</em>. Transwell and endothelial cell tube formation assay were used to measure the effects of miR-<em>145</em> on the invasion and angiogenesis of UM in vitro. The downstream target genes of miR-<em>145</em> were predicted by bioinformatics and confirmed using a luciferase assay. BALB/c nude mice models were established to investigate the mechanisms of miR-<em>145</em> on tumor growth and angiogenesis in vivo. Group data comparisons were performed using analysis of Student's t test. A two-tailed P < 0.05 was considered as statistically significant.

<strong class="sub-title"> Results: </strong> The results of Western blotting analysis indicated that the expressions of N-RAS (1.10 ± 0.35 vs. 0.41 ± 0.36, t = 3.997, P = 0.012), p-AKT (1.16 ± 0.22 vs. 0.57 ± 0.03, t = 7.05, P = 0.001), and VEGF (0.97 ± 0.32 vs. 0.45 ± 0.21, t = 3.314, P = 0.008) in UM tumor tissues were significantly higher than those in normal uveal tissue. Luciferase assay demonstrated N-RAS and VEGF as downstream targets of miR-<em>145</em>. Moreover, tube formation assay revealed that miR-<em>145</em>-transfected human microvascular endothelial cell line formed shorter tube length (36.10 ± 1.51 mm vs. 42.91 ± 0.94 mm, t = 6.603, P = 0.003) and less branch points (350.00 ± 19.97 vs. 406.67 ± 17.62, t = 3.685, P = 0.021) as compared with controls. In addition, the numbers of invaded MUM-2B and OCM-1 cells with miR-<em>145</em> overexpression were significantly lower than the controls (35.7 ± 3.3 vs. 279.1 ± 4.9, t = 273.75, P < 0.001 and 69.5 ± 4.4 vs. 95.6 ± 4.7, t = 21.27, P < 0.001, respectively). In vivo, xenografts expressing miR-<em>145</em> had smaller sizes (miR-<em>145</em> vs. miR-scr, 717.41 ± 502.62 mmvs. 1694.80 ± 904.33 mm, t = 2.314, P = 0.045) and lower weights (miR-<em>145</em> vs. miR-scr, 0.74 ± 0.46 g vs. 1.65 ± 0.85 g, t = 2.295, P = 0.045).

<strong class="sub-title"> Conclusion: </strong> Our results indicated that miR-<em>145</em> is an important tumor suppressor and the inhibitory strategies against N-RAS/VEGF signaling pathway might be potential therapeutic applications for UM in the future.

Currently, at least five different mRNA species encoding <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em>-A (VEGF-A) have been characterized. These variants result from alternative splicing of the VEGF-A transcript and encode human isoforms of VEGF protein of 121, <em>145</em>, 165, 189, and 206 amino acids. In the rat, a similar profile of VEGF-A splice variants has been described, each encoding one fewer amino acid than the human species. Studies of mammalian tissues have shown that these mRNA isoforms vary in abundance. Whereas VEGF 188/189, 164/165, and 120/121 (rat/human, respectively) are the predominant forms expressed in most tissues and cells examined, VEGF 144/<em>145</em> and 205/206 are rare variants. Previously, VEGF 144/<em>145</em> had been detected only in placental and uterine tissues and endometrial carcinoma cell lines, whereas VEGF 205/206 was detected only in fetal liver and placenta. Using an RT-PCR technique, cDNA cloning, and sequencing, we have detected and confirmed the presence of mRNA encoding VEGF 144/<em>145</em> and 205/206 in both adult rat lung and penis. Therefore, these low-abundance VEGF splice variants have a more diverse expression pattern than originally predicted.

<AbstractText>Diet is thought to modulate inflammation. This study shows no relationships between the dietary inflammatory index (DII) and biomarkers of inflammation or bone after adjusting for covariates. Monocyte chemoattractant protein-1 was inversely associated with peripheral tibia cortical thickness and prospective childhood studies should be conducted to better understand this relationship and to determine if there are long-term consequences in adulthood.</AbstractText><AbstractText>Examine the relationships between the DII-scores and bone and biomarkers of inflammation in 290 adolescents, ages 9-13 years.</AbstractText><AbstractText>DII-scores were calculated from 3-day diet records and categorized into tertiles, low (< - 1.34), medium (- 1.34 to 1.41), and high (> 1.41) inflammation. Radius and tibia bone were assessed via peripheral quantitative computed tomography (Stratec XCT 2000) at the 66% site relative to the distal <em>growth</em> plate. Fasting serum was measured for tumor necrosis <em>factor</em> alpha (TNF-α), interleukin-6 (IL-6), <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> (VEGF), and monocyte chemoattractant protein-1 (MCP-1). The relationships between DII-scores and bone and biomarkers of inflammation were assessed using bivariate and partial correlations adjusting for sexual maturation, sex, race, muscle cross-sectional area, and height. ANOVA/ANCOVA models were used to compare DII-tertiles with dependent variables.</AbstractText><AbstractText>DII-scores were negatively associated with tibia trabecular area (TtAr; r = - .141, P = .019), periosteal perimeter (PsPM; r = - .<em>145</em>, P = .016), endosteal perimeter (r = - .<em>145</em>, P = .016), strength strain index (SSI; r = - .129, P = .032), and radius TtAr (r = - .140, P = .020), PsPM (r = -.138, P = .027) and SSI (r = -.131, P = .036) but nullified when adjusting for covariates. Tibia PsPM was higher in the low DII group compared to the medium (P = .050) and high (P = .046) groups but nullified after controlling for covariates. DII-scores were not associated with TNF-α, VEGF, or IL-6, but were associated with MCP-1 only in the unadjusted model (r = .125, P = .042). In the adjusted model, MCP-1 was inversely associated with tibia cortical thickness (r = -.150 P = .030).</AbstractText><AbstractText>The DII-scores were not related to biomarkers of inflammation or bone; however, the biomarker of inflammation, MCP-1 was negatively associated with tibia CtTh. Future prospective pediatric studies should be conducted to better understand this relationship and determine if there are long-term implications in adulthood.</AbstractText>

BACKGROUND

Surgery and anesthesia have been linked to postoperative cognitive disturbance and increased risk of Alzheimer's disease. It is not clear by which mechanisms this increased risk for cognitive disease is mediated. Further, amyloid β production has been suggested to depend on the sleep-wake cycle and neuronal activity. The aim of the present study was to examine if cerebrospinal fluid (CSF) concentrations of a number of biomarkers for Alzheimer's disease-related processes, including amyloid β, neuronal injury, and inflammation, changed over time during intravenous anesthesia in surgical patients.

METHODS

We included patients scheduled for hysterectomy via laparotomy during general anesthesia with intravenous propofol and remifentanil. CSF samples were obtained before, during, and after surgery (5 h after induction) and tested for 27 biomarkers. Changes over time were tested with linear mixed effects models.

RESULTS

A total of 22 patients, all females, were included. The mean age was 50 years (± 9 SD). The mean duration of the anesthesia was <em>145</em> min (± 40 SD). Interleukin (IL)-6, IL-8, monocyte chemoattractant protein 1, and <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> A increased over time. IL-15 and IL-7 decreased slightly over time. Macrophage inflammatory protein 1β and placental <em>growth</em> <em>factor</em> also changed significantly. There were no significant effects on amyloid β (Aβ) or tau biomarkers.

CONCLUSIONS

Surgery and general anesthesia with intravenous propofol and remifentanil induce, during and in the short term after the procedure, a neuroinflammatory response which is dominated by monocyte attractants, without biomarker signs of the effects on Alzheimer's disease pathology or neuronal injury.

The objective of this cross-sectional study was to examine the potential of peri-implant crevicular fluid (PICF) analytes to discriminate between peri-implant health and disease using a multi-biomarker approach.

We collected PICF samples from the mesio-buccal site of every implant (n = <em>145</em>) from 52 subjects with peri-implantitis and measured the levels of 20 biomarkers using Luminex. We grouped implants and subjects based on the clinical characteristic of the sampled sites and implants into: healthy sites from healthy implants (HH), diseased sites from diseased implants (DD) and healthy sites from diseased implants (HD). The significance of the differences between the HH and DD groups was determined using general linear models controlling for false discovery rate. We used logistic regression to determine the best multi-biomarker models that could distinguish HH from DD subjects and HH from HD subjects.

There were statistically significant differences between HH and DD groups for 12/20 biomarkers. Logistic regression resulted in a 6-biomarker model (Flt-3L, GM-CSF, IL-10, sCD40L, IL-17 and TNFα) that discriminated HH from DD subjects (AUC = 0.93) and a 3-biomarker model (IL-17, IL-1ra and vascular endothelial growth factor) that distinguished HH from DD subjects (AUC = 0.90).

PICF biomarkers might help discriminate peri-implant health from disease.

Angiotensin 1-7 (Ang1-7) is an endogenous bioactive component of the renin-angiotensin system (RAS). In addition to its cardio<em>vascular</em> properties, its anti-proliferative and anti-angiogenic traits are believed to play important roles in carcinogenesis. The present study examines the influence of Ang1-7 on processes associated with development and progression of prostate cancer cells. Our findings indicate that while Ang1-7 (1 nM; 48 h) can effectively reduce cell proliferation in DU-<em>145</em>, it can induce a significant decrease in the expression of MKI67 in LNCaP. In both cell lines we also observed a reduction in colony size in soft agar assay. A various changes in gene expression were noted after exposure to Ang1-7: those of anti- and pro-apoptotic agents and the NF-kB family of transcription <em>factors</em>, as well as mesenchymal cell markers and <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> A (VEGFA). In addition, Ang1-7 was found to modulate cell adhesion and matrix metallopeptidase (MMP) activity. Changes were also observed in the levels of angiotensin receptors and sex steroid hormone receptors. Ang1-7 reduced the levels of estrogen receptor alpha gene (ESR1) and increased the expression of estrogen receptor beta gene (ESR2) in all prostate cancer cells; it also up-regulated androgen receptor (AR) expression in androgen-sensitive cells but contradictory effect was observed in androgen- irresponsive cell lines. In summary, the results confirm the existence of complex network between the various elements of the local RAS and the molecular and cellular mechanisms of prostate cancerogenesis. The response of cancer cells to Ang1-7 appears to vary dependently on the dose and time of incubation as well as the aggressiveness and the hormonal status of cells.

The aim of the study was to examine the effect of previous pregnancies and classical cardio<em>vascular</em> risk <em>factors</em> on <em>vascular</em> <em>endothelial</em> function in a group of 264 young and middle-aged women 3 to 11 years postpartum. We examined micro<em>vascular</em> functions by peripheral arterial tonometry and EndoPAT 2000 device with respect to a history of gestational hypertension, preeclampsia, fetal <em>growth</em> restriction, the severity of the disease with regard to the degree of clinical signs and delivery date. Besides, we compared Reactive Hyperemia Index (RHI) values and the prevalence of <em>vascular</em> <em>endothelial</em> dysfunction among the groups of women with normal and abnormal values of BMI, waist circumference, systolic and diastolic blood pressures, heart rate, total serum cholesterol levels, serum high-density lipoprotein cholesterol levels, serum low-density lipoprotein cholesterol levels, serum triglycerides levels, serum lipoprotein A levels, serum C-reactive protein levels, serum uric acid levels, and plasma homocysteine levels. Furthermore, we determined the effect of total number of pregnancies and total parity per woman, infertility and blood pressure treatment, presence of trombophilic gene mutations, current smoking of cigarettes, and current hormonal contraceptive use on the <em>vascular</em> <em>endothelial</em> function. We also examined the association between the <em>vascular</em> <em>endothelial</em> function and postpartum whole peripheral blood expression of microRNAs involved in pathogenesis of cardio<em>vascular</em>/cerebro<em>vascular</em> diseases (miR-1-3p, miR-16-5p, miR-17-5p, miR-20a-5p, miR-20b-5p, miR-21-5p, miR-23a-3p, miR-24-3p, miR-26a-5p, miR-29a-3p, miR-92a-3p, miR-100-5p, miR-103a-3p, miR-125b-5p, miR-126-3p, miR-130b-3p, miR-133a-3p, miR-143-3p, miR-<em>145</em>-5p, miR-146a-5p, miR-155-5p, miR-181a-5p, miR-195-5p, miR-199a-5p, miR-210-3p, miR-221-3p, miR-342-3p, miR-499a-5p, and miR-574-3p). A proportion of overweight women (17.94% and 20.59%) and women with central obesity (18.64% and 21.19%) had significantly lower RHI values at 10.0% false positive rate (FPR) both before and after adjustment of the data for the age of patients. At 10.0% FPR, a proportion of women with <em>vascular</em> <em>endothelial</em> dysfunction (RHI ≤ 1.67) was identified to have up-regulated expression profile of miR-1-3p (11.76%), miR-23a-3p (17.65%), and miR-499a-5p (18.82%) in whole peripheral blood. RHI values also negatively correlated with expression of miR-1-3p, miR-23a-3p, and miR-499a-5p in whole peripheral blood. Otherwise, no significant impact of other studied <em>factors</em> on <em>vascular</em> <em>endothelial</em> function was found. We suppose that screening of these particular microRNAs associated with <em>vascular</em> <em>endothelial</em> dysfunction may help to stratify a highly risky group of young and middle-aged women that would benefit from early implementation of primary prevention strategies. Nevertheless, it is obvious, that <em>vascular</em> <em>endothelial</em> dysfunction is just one out of multiple cardio<em>vascular</em> risk <em>factors</em> which has only a partial impact on abnormal expression of cardio<em>vascular</em> and cerebro<em>vascular</em> disease associated microRNAs in whole peripheral blood of young and middle-aged women.

Angiogenesis is an indispensible process for tumor <em>growth</em> and metastasis. Anti-angiogenesis based therapy is one of the most promising treatments for inhibiting cancer progression. Through the exploration of inhibitors of <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> receptor (VEGFR)-2, deemed as the major angiogenesis pathway, pazopanib was found as a small molecular pan-VEGFR and pan-platelet-derived <em>growth</em> <em>factor</em> receptor (PDGFR) inhibitor, with suitable pharmacodynamic and pharmacokinetic parameters to be an oral drug. In an vitro study, pazopanib exerted anti-tumor effect through mechanisms including the Raf-MAPK/ERK (MEK)-extracellular signal-regulated kinase (ERK) pathway, and directly targeted on v-raf murine sarcoma viral oncogene homolog B (B-raf) as well. It inhibited the proliferation of cell lines, such as DU-<em>145</em> and HRC-45 in hepatocellular carcinoma, through mechanisms like "cell cycle arrest." In vivo xenograft studies and phase I/II clinical trials revealed a series of plasma cytokine and angiogenic <em>factors</em>, such as interleukin (IL)-6, IL-12, hepatocyte <em>growth</em> <em>factor</em> (HGF), and soluble VEGFR2, which have significant association with clinical curative effect. Pazopanib has been shown to be effective in solid tumors and some hematological malignancies. Future studies should focus on the exploration of biomarkers to screen sensitive patients and concomitant or metronomic dosage with other kinds of medicines.

BACKGROUND

Ranibizumab is an inhibitor of vascular endothelial growth factor-A (anti-VEGF) approved for the treatment of neovascular age-related macular degeneration (nAMD). The treat and extend (T&E) regimen can potentially reduce the burden of clinic visits compared with a pro re nata (PRN) regimen. Retrospective, interim analyses of clinical effectiveness, treatment and resource use patterns were conducted using real-world data in England and Wales from the TERRA study.

METHODS

Two cohorts, those switching from a PRN to a T&E regimen ('prior PRN') and those initiating ranibizumab on the T&E regimen as their first anti-VEGF therapy ('anti-VEGF-naïve') were enrolled in TERRA. Retrospective clinical assessments were gathered from medical records, while resource use patterns were collected via an operating cost questionnaire completed by each study site.

RESULTS

At the interim analysis cut-off date (15 November 2016), 11 sites had enrolled 145 patients (prior PRN: n = 110; anti-VEGF-naïve: n = 35). Mean change from baseline (date of first injection) in visual acuity and central subfield retinal thickness to 12 months was +7.6 Early Treatment Diabetic Retinopathy Study letters [95% confidence interval (CI) 2.8, 12.4; p = 0.003; n = 27] and -67.7 μm (95% CI -106.5, -28.9; p = 0.001, n = 29), respectively, in the anti-VEGF-naïve cohort. Most T&E clinics were run as one-stop services (same-day monitoring and injection), whereas 4/10 PRN clinics were run as two-stop services (monitoring and injection on different days). In general, one-stop clinics used less staff resources and were likely to be shorter in duration for healthcare providers than the cumulative time spent for two-stop clinics.

CONCLUSIONS

This is the first real-world observational study conducted in England and Wales demonstrating the effectiveness of the ranibizumab T&E regimen in anti-VEGF-naïve patients. T&E is compatible with one-stop clinic services, which these real-world data suggest to be less resource intensive than two-stop clinic services, possibly providing a dosing regimen beneficial to both patients and resource burden in UK clinical practice.

BACKGROUND

Novartis Pharmaceuticals UK Limited.

Epithelial ovarian cancer (EOC) is a lethal gynaecological neoplasm characterized by rapid <em>growth</em> and angiogenesis. Nerve <em>growth</em> <em>factor</em> (NGF) and its high affinity receptor tropomyosin receptor kinase A (TRKA) contribute to EOC progression by increasing the expression of c-MYC, survivin and <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> (VEGF) along with a decrease in microRNAs (miR) 23b and <em>145</em>. We previously reported that metformin prevents NGF-induced proliferation and angiogenic potential of EOC cells. In this study, we sought to obtain a better understanding of the mechanism(s) by which metformin blocks these NGF-induced effects in EOC cells. Human ovarian surface epithelial (HOSE) and EOC (A2780/SKOV3) cells were stimulated with NGF and/or metformin to assess the expression of c-MYC, β-catenin, survivin and VEGF and the abundance of the tumor suppressor miRs 23b and <em>145</em>. Metformin decreased the NGF-induced transcriptional activity of MYC and β-catenin/T-cell <em>factor</em>/lymphoid enhancer-binding <em>factor</em> (TCF-Lef), as well as the expression of c-MYC, survivin and VEGF in EOC cells, while it increased miR-23b and miR-<em>145</em> levels. The preliminary analysis of ovarian biopsies from women users or non-users of metformin was consistent with these in vitro results. Our observations shed light on the mechanisms by which metformin may suppress tumour <em>growth</em> in EOC and suggest that metformin should be considered as a possible complementary therapy in EOC treatment.

Keywords: NGF; VEGF; c-MYC; epithelial ovarian cancer; metformin; survivin; β-catenin.

MicroRNAs (miRNAs) regulate gene expression; many of them act in the retinal pigment epithelium (RPE), and RPE degeneration is known to be a critical <em>factor</em> in age-related macular degeneration (AMD). Repeated injections with anti-VEGFA (<em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> A) are the only effective therapy in wet AMD. We investigated the correlation between the expression of 18 miRNAs involved in the regulation of the VEGFA gene in serum of 76 wet AMD patients and 70 controls. Efficacy of anti-VEGFA treatment was evaluated by counting the number of injections delivered up to 12 years. In addition, we compared the relative numbers of deaths in patient with AMD and control groups. We observed a decreased expression of miR-34-5p, miR-126-3p, miR-<em>145</em>-5p and miR-205-5p in wet AMD patients as compared with controls. These miRNAs are involved in the regulation of angiogenesis, cytoprotection and protein clearance. No miRNA was significantly correlated with the treatment outcome. Wet AMD patients had greater mortality than controls, and their survival was inversely associated with the number of anti-VEGFA injections per year. No association was observed between miRNA expression and mortality. Our study emphasizes the need to clarify the role of miRNA regulation in AMD pathogenesis.

Sphingosine kinase 2 (SphK2) is known to phosphorylate the nuclear sphingolipid metabolite to generate sphingosine-1-phosphate (S1P). Nuclear S1P is involved in epigenetic regulation of gene expression; however, the underlying mechanisms are not well understood. In this work, we have identified the role of nuclear S1P and SphK2 in regulating hypoxia-responsive master transcription <em>factors</em> hypoxia-inducible <em>factor</em> (HIF)-1α/2α, and their functions in breast cancer, with a focus on triple-negative breast cancer (TNBC). We have shown SphK2 is associated with HIF-1α in protein complexes, and is enriched at the promoters of HIF target genes, including <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> (VEGF), where it enhances local histone H3 acetylation and transcription. S1P specifically binds to the PAS domains of HIF-1α. SphK2, and HIF-1α expression levels are elevated in metastatic estrogen receptor-positive (ER+) and TNBC clinical tissue specimens compared to healthy breast tissue samples. To determine if S1P formation in the nucleus by SphK2 is a key regulator of HIF functions, we found using a preclinical TNBC xenograft mouse model, and an existing selective SphK2 inhibitor K-<em>145</em>, that nuclear S1P, histone acetylation, HIF-1α expression, and TNBC tumor <em>growth</em> were all reduced in vivo. Our results suggest that S1P and SphK2 in the nucleus are linked to the regulation of HIF-1α/2α functions associated with breast cancer progression, and may provide potential therapeutic targets.

Hypoxia-inducible <em>factor</em>-1α (HIF-1α) can control a transcriptional <em>factor</em> forkhead box P3 (Foxp3) protein expression in T lymphocyte differentiation through proteasome-mediated degradation. In this study, we unveil a reverse regulatory mechanism contributing to bladder cancer progression; Foxp3 expression attenuates HIF-1α degradation. We first demonstrated that Foxp3 expression positively correlates with the metastatic potential in T24 cells and can increase the expression of HIF-1α-target genes, such as <em>vascular</em> <em>endothelial</em> <em>growth</em> <em>factor</em> (VEGF) and glucose transporter (GLUT). Foxp3 protein can bind with HIF-1α, particularly under hypoxia. In vivo ubiquination assay demonstrated that Foxp3 can decrease HIF-1α degradation in a dose-dependent manner. Knocking-down of Foxp3 expression blocks in vivo tumor <em>growth</em> in mice and prolongs mice's survival, which is associated with von Willebrand <em>factor</em> expression. Thirty-three of <em>145</em> (22.8 %) bladder tumors exhibit Foxp3 expression. Foxp3 expression is an independent predictor for disease progression in superficial bladder cancer patients (p = 0.032), associated with less number of intratumoral CD8+ lymphocyte. The metaanalysis from 2 published datasets showed Foxp3 expression is positively associated with GLUT-4,-9, and VEGF-A, B-, D expression. This reverse post-translational regulation of HIF-1α protein by Foxp3 provides a new potential target for developing new therapeutic strategy for bladder cancer.