Contribution of MLH1 constitutional methylation for Lynch syndrome diagnosis in patients with tumor MLH1 downregulation

Abstract

Constitutional epimutation of the two major mismatch repair genes, MLH1 and MSH2, has been identified as an alternative mechanism that predisposes to the development of Lynch syndrome. In the present work, we aimed to investigate the prevalence of MLH1 constitutional methylation in colorectal cancer (CRC) patients with abnormal expression of the MLH1 protein in their tumors. In a series of 38 patients who met clinical criteria for Lynch syndrome genetic testing, with loss of MLH1 expression in the tumor and with no germline mutations in the MLH1 gene (35/38) or with tumors presenting the BRAF p.Val600Glu mutation (3/38), we screened for constitutional methylation of the MLH1 gene promoter using methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) in various biological samples. We found four (4/38; 10.5%) patients with constitutional methylation in the MLH1 gene promoter. RNA studies demonstrated decreased MLH1 expression in the cases with constitutional methylation when compared with controls. We could infer the mosaic nature of MLH1 constitutional hypermethylation in tissues originated from different embryonic germ layers, and in one family we could show that it occurred de novo. We conclude that constitutional MLH1 methylation occurs in a significant proportion of patients who have loss of MLH1 protein expression in their tumors and no MLH1 pathogenic germline mutation. Furthermore, we provide evidence that MLH1 constitutional hypermethylation is the molecular mechanism behind about 3% of Lynch syndrome families diagnosed in our institution, especially in patients with early onset or multiple primary tumors without significant family history.

This work aimed to investigate the prevalence of MLH1 constitutional methylation in colorectal cancer patients with abnormal expression of the MLH1 protein in their tumors and without germline mutations. We provide evidence that MLH1 constitutional hypermethylation is the molecular mechanism behind about 3% of Lynch syndrome families, most likely in patients with early onset or multiple primary tumors without significant family history.

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Impact of preoperative anemia on outcomes in patients undergoing curative resection for gastric cancer: a single-institution retrospective analysis of 2163 Chinese patients

Abstract

We sought to evaluate whether preoperative anemia was an important determinant of survival in gastric cancer (GC). A single institution cohort of 2163 GC patients who underwent curative resection were retrospectively analyzed. Anemia was defined as a preoperative hemoglobin level <120 g/L in males and <110 g/L in females. Overall survival (OS) was analyzed using the Kaplan–Meier method, and a multivariate Cox proportional hazards model was performed to identify the independent prognostic factor. Anemic patients had a poorer OS compared with nonanemic patients after resection for tumor–nodes–metastasis (TNM) stage III tumors (5-year OS rate: 32.2% vs. 45.7%, P < 0.001) but not stage I (P  =  0.480) or stage II (P  =  0.917) tumors. Multivariate analysis revealed that preoperative anemia was an independent prognostic factor in TNM stage III (hazard ratio [HR], 1.771; 95% CI, 1.040–3.015; P = 0.035). In a stage-stratified analysis, preoperative anemia was still independently associated with OS in TNM stages IIIa through IIIc (P < 0.001, P = 0.075, and P = 0.012, respectively), though the association was only marginal in stage IIIb. Of note, preoperative mild anemia had a similar prognostic value in TNM stage III GC. Furthermore, preoperative anemia was significantly associated with more perioperative transfusions, postoperative complications and several nutritional-based indices, including the prognostic nutritional index (PNI), preoperative weight loss and performance status (all P < 0.05). Preoperative anemia, even mild anemia, was an important predictor of postoperative survival for TNM stage III GC.

Preoperative anemia, even mild anemia, is a useful predictor of outcomes in tumor–nodes–metastasis stage III gastric cancer.

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Downregulation of CSN6 attenuates papillary thyroid carcinoma progression by reducing Wnt/β-catenin signaling and sensitizes cancer cells to FH535 therapy

Abstract

The incidence of thyroid cancer has increased worldwide at a rate higher than that of any other cancer. CSN6 is overexpressed in many types of cancers, and such expression is linked to oncogenic activity. However, the detailed biological functions of CSN6 in papillary thyroid cancer (PTC) have not been well characterized. We investigated CSN6 expression in PTC specimens and cell lines. We used short-hairpin RNA-mediated gene silencing to explore the biological effects of CSN6 depletion in PTC cells. The combined effects of CSN6 silencing and FH535 therapy were assessed in terms of cell viability. The mechanism by which CSN6 regulated β-catenin expression was also analyzed. CSN6 levels were determined by real-time polymerase chain reaction (PCR) (mRNA), Western blotting, and immunochemistry (protein). The CCK-8 and migration assays and orthotopic xenograft transplantation were used to investigate the biological effects of CSN6. We assessed the combined effects of CSN6 silencing and FH535 on cell viability in vitro. We also analyzed the relationship between the CSN6 level and clinical pathological status. CSN6 was overexpressed in human PTCs, and loss of CSN6 attenuated tumor proliferation and migration both in vitro and in vivo. CSN6 stabilized β-catenin and facilitated the epidermal-to-mesenchymal transition (EMT) in PTC cells. CSN6 positively regulated β-catenin expression in a β-Trcp-dependent manner and triggered expression of several EMT-related genes regulated by β-catenin. CSN6 silencing sensitized PTC cells to FH535 therapy via downregulation of the Wnt/β–catenin signaling pathway. Finally, in PTC patients, the level of CSN6 was significantly (inversely) correlated with tumor size, the presence of multifocal lesions, and TNM stage. CSN6 overexpression in PTC is a strong indicator of enhanced tumor aggressiveness. CSN6 promotes PTC progression by inducing the EMT. CSN6 knockdown sensitizes PTC cells to FH535 therapy via downregulation of the Wnt/β–catenin signaling pathway.

CSN6 was overexpressed in human PTCs, and loss of CSN6 attenuated tumor proliferation and migration. In PTC patients, the level of CSN6 was significantly (inversely) correlated with tumor size, the presence of multifocal lesions, and TNM stage.

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Clinical outcomes of carbon ion radiotherapy with concurrent chemotherapy for locally advanced uterine cervical adenocarcinoma in a phase 1/2 clinical trial (Protocol 1001)

Abstract

We conducted a phase 1/2 study to evaluate the efficacy and safety of carbon ion radiotherapy (C-ion RT) with concurrent chemotherapy for locally advanced uterine cervical adenocarcinoma. Thirty-three patients were enrolled between April 2010 and March 2014. Treatment consisted of C-ion RT with concurrent weekly cisplatin at a dose of 40 mg/m². In the phase 1 component, the total dose was escalated from 68.0 Gy (relative biological effectiveness [RBE]) to 74.4 Gy (RBE) to determine the maximum tolerated dose of C-ion RT. In the phase 2 component, the efficacy and safety of C-ion RT with concurrent chemotherapy were evaluated using the dose determined in the phase 1 component. The median follow-up duration was 30 months. Two patients did not receive chemotherapy because of anemia or leukocytopenia immediately prior to commencing treatment; 31 patients were analyzed. None of the patients developed dose-limiting toxicities. The recommended dose (RD) was determined to be 74.4 Gy (RBE). In the phase 2 component, two patients developed Grade 3–4 toxicities in the gastrointestinal tract, due to repeated laser coagulation or peritonitis caused by appendicitis. In the patients treated with the RD, the 2-year local control, progression-free survival, and overall survival rates were 71%, 56%, and 88%, respectively. C-ion RT with concurrent weekly cisplatin was well tolerated in patients with locally advanced uterine cervical adenocarcinoma. Our findings support further investigations into the efficacy of this strategy.

The efficacy and safety of carbon ion radiotherapy with concurrent chemotherapy was evaluated in patients with locally advanced uterine cervical adenocarcinoma. This strategy was well tolerated in the majority of patients.

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Mechanism underlying the negative effect of prostate volume on the outcome of extensive transperineal ultrasound-guided template prostate biopsy

Abstract

Previous studies have indicated a possible relationship between increased prostate volume (PV) and decreased biopsy yield, although the mechanism involved is unclear. We evaluated 1650 patients who underwent template biopsy. The distribution of 993 cancer lesions in 302 prostatectomy specimens was compared with the biopsy data to determine whether each lesion was detected. A receiver operating characteristic (ROC) model was used to determine the diagnostic accuracy of prostate-specific antigen (PSA) and related markers. A medical record number (MRN) was used as a negative control. The cancer positive rate did not change as PSA increased in patients with PV ≥50 mL (P = 0.466), although it increased as PSA increased in patients with PV<50 mL (P = 0.001). The detection rate of cancer lesions decreased as the diameter of the lesions decreased (P = 0.018), but remained unchanged with respect to PV. The diameters of the maximum lesions in patients with PV ≥ 50 mL were significantly smaller than those in patients with PV<50 mL (P = 0.003). In patients with PV ≥ 50 mL, the areas under the ROC curves for PSA-related markers did not differ significantly from that for MRN, although they were significantly greater than that for MRN in patients with PV<50 mL (P < 0.001). These results suggest that an increase in PV is associated with a decrease in size and detectability of cancer lesions resulting in a decrease in biopsy yield. Loss of diagnostic accuracy of markers in patients with PV ≥ 50 mL indicates a decrease in serum levels of PSA produced by prostate cancer, which suggests growth inhibition of the cancer.

The currents study indicates that the diameters of maximum cancer lesions in patients with prostate volume ≥ 50 mL were significantly smaller than those in patients with prostate volume <50 mL. Loss of diagnostic accuracy of markers in patients with prostate volume ≥ 50mL indicates a decrease in serum levels of PSA produced by prostate cancer, which suggests growth inhibition of the cancer.

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Nuclear division cycle 80 promotes malignant progression and predicts clinical outcome in colorectal cancer

Abstract

Colorectal cancer (CRC) is a common human malignancy worldwide and increasing studies have attributed its malignant progression to abnormal molecular changes in cancer cells. Nuclear division cycle 80 (NDC80) is a newly discovered oncoprotein that regulates cell proliferation and cycle in numerous malignancies. However, its clinical significance and biological role in CRC remain unclear. Therefore, in this study, we firstly analyze its expression in a retrospective cohort enrolling 224 CRC patients and find its overexpression is significantly correlated with advanced tumor stage and poor prognosis in CRC patients. In addition, our result reveals it is an independent adverse prognostic factor affecting CRC-specific and disease-free survival. The subgroup analysis indicates NDC80 expression can stratify the clinical outcome in stage II and III patients, but fails in stage I and IV patients. In cellular assays, we find knockdown of NDC80 dramatically inhibits the proliferative ability, apoptosis resistance, cell cycle progression, and clone formation of CRC cells in vitro. Using xenograft model, we further prove knockdown of NDC80 also inhibits the tumorigenic ability of CRC cells in vivo. Finally, the microarray analysis is utilized to preliminarily clarify the oncogenic molecular mechanisms regulated by NDC80 and the results suggest it may promote CRC progression partly by downregulating tumor suppressors such as dual specificity phosphatase 5 and Forkhead box O1. Taken together, our study provides novel evidences to support that NDC80 is not only a promising clinical biomarker but also a potential therapeutical target for CRC precise medicine.

In summary, our results indicate that high NDC80 expression is correlated with advanced tumor stage and unfavorable prognosis in CRC patients. In addition, we also find NDC80 drives malignant progression of CRC cells partly by inactivating DUSP5 and FOXO1. Taken together, these evidences suggest NDC80 is not only a promising clinical biomarker for patient management, but also a potential therapeutical target for CRC diagnosis and treatment.

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Editorial: tofacitinib and biologics for moderate-to-severe ulcerative colitis—what is best in class?

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Editorial: vedolizumab as a treatment and cause of extra-intestinal manifestations of inflammatory bowel disease

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Editorial: tofacitinib and biologics for moderate-to-severe ulcerative colitis—what is best in class? Authors’ reply

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Editorial: direct-acting antivirals significantly improve quality of life in patients with hepatitis C virus infection—Author's reply

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Issue Information

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Editorial: getting bullish about portal hypertension—chronic treatment with oral taurine? Author's reply

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Letter: addition of azathioprine to infliximab maintenance therapy in patients with anti-drug antibodies and subclinical inflammation

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Editorial: direct-acting antivirals significantly improve quality of life in patients with hepatitis C virus infection

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This article is linked to Younossi et al and Younossi papers. To view these articles visit http://ift.tt/2DEU211 and http://ift.tt/2DfjKIc.

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Editorial: getting bullish about portal hypertension—chronic treatment with oral taurine?

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This article is linked to Schwarzer et al and Ferlitsch papers. To view these articles visit http://ift.tt/2DbcFIO and http://ift.tt/2DExoWq.

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Letter: more studies are needed to elucidate the impact of HBV/HCV coinfection on cirrhosis and its consequences

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Letter: more studies are needed to elucidate the impact of HBV/HCV coinfection on cirrhosis and its consequences—Authors’ reply

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This article is linked to Pol et al and Huang et al papers. To view these articles visit http://ift.tt/2DEGqCV and http://ift.tt/2DdI65v.

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Assessment of potential risk factors for breast cancer in a population in Southern Brazil

Abstract

Purpose

The aim of this study is to assess potential risk factors for breast cancer in a population in Southern Brazil and build a multivariate logistic model using these factors for breast cancer risk prediction.

Methods

A total of 4242 women between 40 and 69 years of age without a history of breast cancer were selected at primary healthcare facilities in Porto Alegre and submitted to mammographic screening. They were evaluated for potential risk factors.

Results

In all, 73 participants among the 4242 women had a breast cancer diagnosis during the follow-up of the project (10 years). The multivariate analysis considering all the patients aged 40–69 years showed that older age (OR 1.08, 95% CI 1.04–1.12), higher height (OR 1.04, 95% CI 1.01–1.09), and history of previous breast biopsy (OR 2.66, 95% CI 1.38–5.13) were associated with the development of breast cancer. Conversely, the number of pregnancies (OR 0.87, 95% CI 0.78–0.98) and use of hormone replacement therapy (OR 0.39, 95% CI 0.20–0.75) were considered a protective factor. Additionally, we performed an analysis separating the participants into groups of 40–49 and 50–69 years old, since a risk factor could have a specific behavior in these age groups. No additional risk factors were identified within these age brackets, and some factors lost statistical significance.

Conclusion

The risk prediction model indicates that the following variables should be assessed in this specific population: age, height, having had previous breast biopsies, number of pregnancies, and use of hormone replacement therapy. These findings may help to better understand the causal model of breast cancer in Southern Brazil.

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Unifying control over the body: consciousness and cross-cueing in split-brain patients

Sir,

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SCO2 mutations cause early-onset axonal Charcot-Marie-Tooth disease associated with cellular copper deficiency

Abstract

Recessive mutations in the mitochondrial copper-binding protein SCO2, cytochrome c oxidase (COX) assembly protein, have been reported in several cases with fatal infantile cardioencephalomyopathy with COX deficiency. Significantly expanding the known phenotypic spectrum, we identified compound heterozygous variants in SCO2 in two unrelated patients with axonal polyneuropathy, also known as Charcot-Marie-Tooth disease type 4. Different from previously described cases, our patients developed predominantly motor neuropathy, they survived infancy, and they have not yet developed the cardiomyopathy that causes death in early infancy in reported patients. Both of our patients harbour missense mutations near the conserved copper-binding motif (CXXXC), including the common pathogenic variant E140K and a novel change D135G. In addition, each patient carries a second mutation located at the same loop region, resulting in compound heterozygote changes E140K/P169T and D135G/R171Q. Patient fibroblasts showed reduced levels of SCO2, decreased copper levels and COX deficiency. Given that another Charcot-Marie-Tooth disease gene, ATP7A, is a known copper transporter, our findings further underline the relevance of copper metabolism in Charcot-Marie-Tooth disease.

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Better characterization of vinflunine pharmacokinetics variability and exposure/toxicity relationship to improve its use: analyses from 18 trials

Summary

Aims

Vinflunine is a novel tubulin-targeted inhibitor indicated as a single agent for the treatment of bladder cancers after failure of prior platinum-based therapy. Its pharmacokinetics (PK) and pharmacodynamics (PD) have been independently characterized through several phase I and phase II studies. However, no global pharmacometric analysis had been conducted yet.

Methods

Vinflunine concentrations and safety data from 18 phase I and phase II studies were used to conduct population PK and PK/PD analysis, using Nonmem. A 4-compartment model was used to describe vinflunine PK and several covariates were tested in order to explain inter-individual variability. In terms of PK/PD relationship, a semi-physiologic population PK/PD model was applied to describe time course of absolute neutrophil counts (ANC) after vinflunine administration and logistic regression models were used to test the relationship between vinflunine exposure and toxicities.

Results

Vinflunine clearance is explained by creatinine clearance, BSA and combination with pegylated doxorubicin, leading to a decrease from 28.2 to 25.3% of the inter-individual variability. When vinflunine dose is decreased, simulations of ANC time course (via a semi-physiological model) after vinflunine administration show a risk of neutropenia grade 3-4 at cycle 2 always lower than when dose is delayed. As an example, for moderate renal impaired patients, the risk is 42.1% when vinflunine is dosed at 320 mg/m² q4w vs. 23.3% for 280 mg/m² q3w.

Conclusions

We propose for the first time a global comprehensive clinical pharmacological analysis for IV vinflunine that may help drive dose adjustment.

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Pharmacokinetic/pharmacodynamic drug-drug interactions of avatrombopag when co-administered with dual or selective CYP2C9 and CYP3A interacting drugs

Summary

Aims

Avatrombopag, a thrombopoietin receptor agonist, is a substrate of cytochrome P450 (CYP) 2C9 and CYP3A. We assessed three drug-drug interactions of avatrombopag as a victim with dual or selective CYP2C9/3A inhibitors and inducers.

Methods

This was a 3-part, open-label study. Forty-eight healthy subjects received single 20 mg doses of avatrombopag alone or with one of 3 CYP2C9/3A inhibitors or inducers: fluconazole 400 mg once daily for 16 days, itraconazole 200 mg twice daily on Day 1 and 200 mg once daily on Days 2-16, or rifampin 600 mg once daily for 16 days. Pharmacokinetics, pharmacodynamics (platelet count), and safety of avatrombopag were evaluated.

Results

Co-administration of a single 20-mg dose of avatrombopag with fluconazole at steady-state resulted in 2.16-fold increase of AUC of avatrombopag, prolonged t½ (from 19.7 h to 39.9 h) and clinically significant increase in E_max (1.66-fold). Itraconazole had a mild increase on both avatrombopag pharmacokinetics and pharmacodynamics compared to fluconazole. Co-administration of rifampin caused a 0.5-fold decrease in AUC, shortened t½ (from 20.3 h to 9.84 h), and without any impact of E_max. Co-administration with interacting drugs was found to be generally safe and well-tolerated.

Conclusions

The results from co-administration of fluconazole or itraconazole suggest that CYP2C9 plays a predominant role in metabolic clearance of avatrombopag than CYP3A. To achieve comparable platelet count increases when avatrombopag is co-administered with CYP3A and CYP2C9 inhibitors, an adjustment in the dose or duration of treatment is recommended, while co-administration with strong inducers is not currently recommended.

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Blind sequential detection for sparse ISI channels

We present a computationally efficient blind sequential detection method for data transmitted over a sparse intersymbol interference channel. Unlike blind sequential detection methods designed for general chan...

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Cytological features of mixed adenoneuroendocrine carcinoma of the ampulla of Vater: A case report with immunocytochemical analyses

Mixed adenoneuroendocrine carcinoma (MANEC) is defined as a tumor that has morphologically recognizable both adenocarcinoma and neuroendocrine carcinoma components comprising at least 30% of either components. MANEC occurring in the ampulla of Vater is extremely rare, and only 16 cases have been reported in the English language literature. In the present report, we describe the first case of MANEC of the ampulla of Vater with immunocytochemical analyses. An 82-year-old Japanese male was incidentally found to have a tumorous lesion in the ampulla of Vater. Endoscopic ultrasound-fine needle aspiration (EUS-FNA) of the tumor was performed. The Papanicolaou smear demonstrated the presence of different three components. The most dominant component was cohesive clusters of small round cells with round to oval nuclei with powdery chromatin and scant cytoplasm, which corresponded to small cell carcinoma. The second component was an adenocarcinoma, which was composed of irregularly overlapping clusters of tall columnar cells with large round to oval nuclei containing conspicuous nucleoli. The third component was an adenoma, which was comprised of flat cohesive clusters of columnar cells without atypia. Immunocytochemical analyses demonstrated that synaptophysin was expressed in the small round cells, and cdx-2 was expressed in all three components. Accordingly, a cytodiagnosis of MANEC with adenoma component was made. Preoperative diagnosis of ampullary MANEC is difficult. However, this report clearly demonstrates three different components in the EUS-FNA cytological specimen. Therefore, we suggest that cytological examination is a useful method for diagnosis of MANEC of the ampulla of Vater.

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Iron pill aspiration: Cytologic and histologic findings of a potential life-threatening airway injury. A Case report and literature review

Iron pill-induced injury of bronchial mucosa is a complication following accidental aspiration of an iron tablet. Oral iron supplementation is a common therapy, particularly among advanced-age patients, who are more prone to aspiration. However, iron pill aspiration has been rarely reported in the literature, usually under the format of short case reports, with only 32 cases published in the literature. The cytologic features suspicious for this rare but potentially lethal entity have been seldom described. We report a case of a patient diagnosed with iron pill-induced bronchial injury, after oral ferrous sulfate has been prescribed during a hospital admission for pneumonia. In the bronchial washing specimen, a background of necrotic cell debris and acute inflammation involving extracellular golden-brown fibrils positive for iron stains was seen, along with the yeast forms, which, in this clinical context could confirm the iron pill aspiration. Our aim is to highlight the cytology features associated with iron pill aspiration bronchitis, and to review the literature for the histologic, clinical, bronchoscopy, and treatment aspects.

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Stability, integrity, and recovery rate of cellular nucleic acids preserved in a new liquid-based cytology medium

Background

Liquid-based cytology (LBC) has replaced the conventional Papanicolaou test in cervical cancer screening. The cervical swab specimens collected in LBC media can also be used for additional analyses including high-risk HPV (HR-HPV) test, DNA methylation analysis, and HPV E6/E7 mRNA test.

Methods

The stability, integrity, and recovery rate of cellular DNA and RNA after storage at different conditions were evaluated by a quantitative real-time PCR (qPCR) based HR-HPV test, reverse transcription qPCR (RT-qPCR), and agarose gel electrophoresis. Cervical swab specimens collected in a newly developed LBC medium, VersaMedium, and ThinPrep PreservCyt medium were processed on Hologic ThinPrep 5000 instrument.

Results

Cervical exfoliative cells fixed by VersaMedium exhibited good cellular morphology with intact membranes and delineated chromatin structures. Cellular DNA preserved in VersaMedium exhibited high level of stability at both room temperature and 4°C, and remained mostly intact at 4°C for up to 28 days. Cellular RNA preserved in VersaMedium maintained higher level of stability and integrity at 4°C than at room temperature. VersaMedium also showed no apparent adverse effect on the recovery rate of nucleic acids.

Conclusion

In addition to maintaining cellular morphology, when stored at 4°C, VersaMedium preserves cellular nucleic acids and PreservCyt medium without noticeable adverse effects on the recovery rate during purification. Therefore, VersaMedium is an appropriate LBC medium for the collection and preservation of cervical swab specimens. And VersaMedium preserved cellular nucleic acids are of such high quality that they are suitable for HR-HPV qPCR test and RT-qPCR analyses.

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Evidence-based adequacy criteria for instrumented urine cytology using cytospin preparations

Cytospin preparations of instrumented urine cytology specimens with less than 10 urothelial cells or more than 50 urothelial cells/10 hpfs are both associated with significantly increased false negative rates compared to cases with 10-49 urothelial cells/10 hpfs.

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Optimizing outcomes in EGFR mutation-positive NSCLC: which tyrosine kinase inhibitor and when?

Future Oncology, Ahead of Print.


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Endovascular implantation of 125I seed combined with transcatheter arterial chemoembolization for unresectable hepatocellular carcinoma

Future Oncology, Ahead of Print.


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NrF2/ARE and NF-κB pathway regulation may be the mechanism for lutein inhibition of human breast cancer cell

Future Oncology, Ahead of Print.


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Associations of coffee consumption and caffeine intake with mammographic breast density

Abstract

Purpose

Previous studies suggest that coffee and caffeine intake may be associated with reduced breast cancer risk. We investigated the association of coffee and caffeine intake with mammographic breast density by woman's menopausal status and, in postmenopausal women, by hormone therapy (HT).

Methods

This study included 4130 cancer-free women within the Nurses' Health Study and Nurses' Health Study II cohorts. Percent breast density (PD) was measured from digitized film mammograms using a computer-assisted thresholding technique and square root-transformed for the analysis. Average cumulative coffee/caffeine consumption was calculated using data from all food frequency questionnaires preceding the mammogram date. Information regarding breast cancer risk factors was obtained from questionnaires closest to the mammogram date. We used generalized linear regression to quantify associations of regular, decaffeinated, and total coffee, and energy-adjusted caffeine intake with percent density.

Results

In multivariable analyses, decaffeinated coffee was positively associated with PD in premenopausal women (2+ cups/day: β = 0.23, p trend = 0.03). In postmenopausal women, decaffeinated and total coffee were inversely associated with PD (decaffeinated 2+ cups/day: β = − 0.24, p trend = 0.04; total 4+ cups/day: β = − 0.16, p trend = 0.02). Interaction of decaffeinated coffee with menopausal status was significant (p-interaction < 0.001). Among current HT users, regular coffee and caffeine were inversely associated with PD (regular coffee 4+ cups/day: β = − 0.29, p trend = 0.01; caffeine 4th vs. 1st quartile: β = − 0.32, p trend = 0.01). Among past users, decaffeinated coffee was inversely associated with PD (2+ cups/day β = − 0.70, p trend = 0.02).

Conclusions

Associations of decaffeinated coffee with percent density differ by woman's menopausal status. Associations of regular coffee and caffeine with percent density may differ by HT status.

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Genomic Identification and Functional Characterization of Essential Genes in Caenorhabditis elegans

Using combined genetic mapping, Illumina sequencing, bioinformatics analyses, and experimental validation, we identified 60 essential genes from 104 lethal mutations in two genomic regions of C. elegans totalling approximately 14 Mb on chromosome III(mid) and chromosome V(left). Five of the 60 genes had not previously been shown to have lethal phenotypes by RNA interference depletion. By analyzing the regions around the lethal missense mutations, we identified four putative new protein functional domains. Furthermore, functional characterization of the identified essential genes shows that most are enzymes, including helicases, tRNA synthetases, and kinases in addition to ribosomal proteins. Gene Ontology analysis indicated that essential genes often encode for enzymes that conduct nucleic acid binding activities during fundamental processes, such as intracellular DNA replication, transcription, and translation. Analysis of essential gene shows that they have fewer paralogs, encode proteins that are in protein interaction hubs, and are highly expressed relative to non-essential genes. All these essential gene traits in C. elegans are consistent with those of human disease genes. Most human orthologs (90%) of the essential genes in this study are related to human diseases. Therefore, functional characterization of essential genes underlines their importance as proxies for understanding the biological functions of human disease genes.

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Genetic Diversity, Molecular Phylogeny and Selection Evidence of Jinchuan Yak Revealed by Whole-Genome Resequencing

Jinchuan yak, a newly discovered yak breed, not only possesses a large proportion of multi-ribs but also exhibits many good characteristics, such as high meat production, milk yield and reproductive performance. However, information about their overall genetic structure, relationship with yaks in other areas, and possible origins and evolutionary processes is limited. In this study, 7,693,689 high-quality single-nucleotide polymorphisms were identified by resequencing the genome of Jinchuan yak. Principal component and population genetic structure analyses showed that Jinchuan yak was distinguished as an independent population among the domestic yak population. Linkage disequilibrium analysis showed that the decay rate of Jinchuan yak was the lowest in the domestic yak breeds, indicating that the degree of domestication and selection intensity of Jinchuan yak were higher than those of other yak breeds. Combined with archaeological data, we speculated that the origin of domestication of Jinchuan yak was about 6,000 years ago (4,000-10,000 years ago). The quantitative dynamics of population growth history in Jinchuan yak was similar to that of other breeds of domestic and wild yaks but was closer to that of the wild yak. No significant gene exchange between Jinchuan and other domestic yaks occurred. Compared with other domestic yaks, Jinchuan yak possessed 339 significantly and positively selected genes, several of which relate to physiological rhythm, histone and the excellent production characters of Jinchuan yak. Our results provide a basis for the discovery of the evolution, molecular origin, and unique traits of Jinchuan yak.

http://ift.tt/2EOSzVp

Repurposing tin mesoporphyrin as an immune checkpoint inhibitor shows therapeutic efficacy in preclinical models of cancer

Purpose: Unprecedented clinical outcomes have been achieved in a variety of cancers by targeting immune checkpoint molecules. This preclinical study investigates heme oxygenase-1 (HO-1), an immune suppressive enzyme that is expressed in a wide variety of cancers, as a potential immune checkpoint target in the context of a chemotherapy-elicited anti-tumor immune response. We evaluate repurposing tin mesoporphyrin (SnMP), which has demonstrated safety and efficacy targeting hepatic HO in the clinic for the treatment of hyperbilirubinaemia, as an immune checkpoint blockade therapy for the treatment of cancer. Experimental design: SnMP and genetic inactivation of myeloid HO-1 were evaluated alongside 5-fluorouracil in an aggressive spontaneous murine model of breast cancer (MMTV-PyMT). Single-cell RNA sequencing analysis, tumor microarray and clinical survival data from breast cancer patients were used to support the clinical relevance of our observations. Results: We demonstrate that SnMP inhibits immune suppression of chemotherapy-elicited CD8+ T cells by targeting myeloid HO-1 activity in the tumor microenvironment. Microarray and survival data from breast cancer patients reveal that HO-1 is a poor prognostic factor in patients receiving chemotherapy. Single-cell RNA sequencing analysis suggests that the myeloid lineage is a significant source of HO-1 expression, and is co-expressed with the immune checkpoints PD-L1/2 in human breast tumors. In vivo, we therapeutically compared the efficacy of targeting these two pathways alongside immune-stimulating chemotherapy, and demonstrate that the efficacy of SnMP compares favorably to PD-1 blockade in preclinical models. Conclusions: SnMP could represent a novel immune checkpoint therapy, which may improve the immunological response to chemotherapy.

http://ift.tt/2rfhJdK

IDH1/2 mutations sensitize acute myeloid leukemia to PARP inhibition and this is reversed by IDH1/2-mutant inhibitors

Purpose: Somatic mutations in IDH1/2 occur in ~20% of patients with myeloid neoplasms, including acute myeloid leukemia (AML). IDH1/2MUT enzymes produce D-2-hydroxyglutarate (D2HG), which associates with increases in DNA damage and improved responses to chemo/radiotherapy and PARP inhibitors in solid tumor cells. Whether this also holds true for IDH1/2MUT AML is not known. Experimental Design: Well-characterized primary IDH1MUT, IDH2MUT and IDH1/2WT AML cells were analyzed for DNA damage and responses to daunorubicin, ionizing radiation and PARP inhibitors. Results: IDH1/2MUT caused increased DNA damage and sensitization to daunorubicin, irradiation, and the PARP inhibitors olaparib and talazoparib in AML cells. IDH1/2MUT inhibitors protected against these treatments. Combined treatment with a PARP inhibitor and daunorubicin had an additive effect on the killing of IDH1/2MUT AML cells. We provide evidence that the therapy sensitivity of IDH1/2MUT cells was caused by D2HG-mediated downregulation of expression of the DNA damage response gene ATM and not by altered redox responses due to metabolic alterations in IDH1/2MUT cells. Conclusions: IDH1/2MUT AML cells are sensitive to PARP inhibitors as monotherapy but especially when combined with a DNA-damaging agent such as daunorubicin, whereas concomitant administration of IDH1/2MUT inhibitors during cytotoxic therapy decrease the efficacy of both agents in IDH1/2MUT AML. These results advocate in favor of clinical trials of PARP inhibitors either or not in combination with daunorubicin in IDH1/2MUT AML.

http://ift.tt/2mMbZ6H

Intrapulmonary Pharmacokinetics of Lascufloxacin in Healthy Adult Volunteers [PublishAheadOfPrint]

This study was performed to investigate the intrapulmonary penetration of lascufloxacin in humans. Thirty healthy adult male Japanese subjects, allocated into five groups, received lascufloxacin in single oral dose of 75 mg. Bronchoalveolar lavage and blood sampling were performed simultaneously in each subject at 1, 2, 4, 6, or 24 hours after administration, and lascufloxacin concentrations in plasma, epithelial lining fluid, and alveolar macrophages were determined. Lascufloxacin was rapidly distributed to the epithelial lining fluid with T_max of 1 hour, which was identical to that in plasma. The C_max values in plasma, epithelial lining fluid, and alveolar macrophages were 0.576, 12.3, and 21.8 μg/mL, respectively. The corresponding AUC_0-24 values were 7.67, 123, and 325 μg⋅h/mL. The mean drug concentrations in the epithelial lining fluid and alveolar macrophages were much higher than those in plasma at all time points examined, and the average site-to-free plasma concentration ratios fell within the ranges of 57.5 -- 86.4 and 71.0 -- 217, respectively. Drug levels in epithelial lining fluid and alveolar macrophages exceeded the MIC₉₀ values for common respiratory pathogens. (This study was registered at JAPIC under registration number JapicCTI- 142547.)

http://ift.tt/2ritDUg

A Novel Piperazine-Based Drug Lead for Cryptosporidiosis from the Medicines for Malaria Venture Open Access Malaria Box [PublishAheadOfPrint]

Cryptosporidiosis causes life-threatening diarrhea in children under age five, and prolonged diarrhea in immunodeficient people, especially AIDS patients. The standard of care, nitazoxanide, is modestly effective in children and ineffective in immunocompromised individuals. In addition to a need for new drugs, better knowledge of drug properties that drive in vivo efficacy is needed to facilitate drug development. We report identification of a piperazine-based lead compound for Cryptosporidium drug development, MMV665917, and a new pharmacodynamic method used for its characterization. MMV665917 was identified from the Medicines for Malaria Venture Malaria Box, followed by dose response studies, in vitro toxicity studies, and structure activity relationship studies using commercial analogues. Potency against C. parvum Iowa and field isolates, and C. hominis was comparable. Furthermore, unlike nitazoxanide, clofazimine, and paromomycin, MMV665917 appeared to be curative in a chronic NOD SCID gamma mouse model of cryptosporidiosis. MMV665917 was also efficacious in an acute interferon- knockout mouse model. To determine if efficacy in this chronic mouse model might relate to whether compounds are cidal or static for C. parvum, we developed a novel in vitro parasite persistence assay. This assay suggested that MMV665917 was cidal, unlike nitazoxanide, clofazimine, and paromomycin. It also enabled determination of the compound concentration required to maximize the rate of parasite elimination. This time-kill assay can be used to prioritize early-stage Cryptosporidium drug leads, and may aide in planning in vivo efficacy experiments. Collectively, these results identify MMV665917 as a promising lead, and establish a new method for characterizing potential anticryptosporidial agents.

http://ift.tt/2mNMNN3

Early clinical assessment of the antimicrobial activity of finafloxacin compared to ciprofloxacin in sub-sets of microbiologically characterized isolates [PublishAheadOfPrint]

Introduction

Two phase II studies were performed in patients with uncomplicated (uUTI) and complicated urinary tract infections or acute pyelonephritis (cUTI/PN) to compare finafloxacin (300 mg b.i.d, p.o. for uUTI 800 mg q.d., i.v. for cUTI/PN) and ciprofloxacin (250 mg b.i.d, p.o. for uUTI and 400 mg b.i.d., i.v. for cUTI/PN). The early response to the study medications was evaluated in the microbiological-intent-to treat population (mITT) at day 3.

Results

21% of the isolates were ciprofloxacin-resistant, 13.7 % were primed pathogens carrying mutation(s) potentially fostering fluoroquinolone-resistance development, and 7.1 % produced ESBLs. Finafloxacin demonstrated very good early clinical activity with microbiological eradication rates of 88.6% (n=132) compared to 78.7% (n=61) for ciprofloxacin, and 69.6% (n=23) compared to 35.7% (n=14) for ciprofloxacin in patients with ciprofloxacin-resistant, and 94.1% (n=17) compared to 80.0% (n=10) for ciprofloxacin in patients infected with uropathogens primed for fluoroquinolone-resistance uropathogens, and 91.7% (n=11) for finafloxacin compared to 0% for ciprofloxacin in patients infected with ESBL-producers.

Conclusions

Finafloxacin demonstrated an early and rapid activity against uropathogens including fluoroquinolone- and/or multi-resistant or ESBL-producers while ciprofloxacin was less active against this subset of resistant pathogens.

Methods

Susceptibilities of pathogens were quantitated by broth microdilution. Isolates were subgrouped according to their susceptibility-patterns, in particular first-step-quinolone-resistance, quinolone-resistance, and ESBL-production. Eradication was defined as elimination or reduction of study entry pathogens to <10³ CFU/ml in urine culture.

http://ift.tt/2rf7RRg

Identification of Hsp90 inhibitors with anti-Plasmodium activity [PublishAheadOfPrint]

Malaria remains a global health burden partly due to Plasmodium parasite resistance to first-line therapeutics. The molecular chaperone heat shock protein 90 (Hsp90) has emerged as an essential protein for blood stage Plasmodium parasites, but details about its function during malaria's elusive liver stage are unclear. We used target-based screens to identify compounds that bind to Plasmodium falciparum and human Hsp90, which revealed insights into chemotypes with species selective binding. Using cell-based malaria assays, we demonstrate that all identified Hsp90-binding compounds are liver and blood stage Plasmodium inhibitors. Additionally, the Hsp90 inhibitor SNX-0723 synergistically reduces liver stage parasite load in combination with the phosphatidylinositol 3-kinase inhibitor PIK-75. Time course inhibition studies with the Hsp90 inhibitors and expression analysis support a role for Plasmodium Hsp90 in late liver stage parasite development. Our results suggest that Plasmodium Hsp90 is essential to liver and blood stage parasite infections and highlight an attractive route to develop species selective PfHsp90 inhibitors that could act synergistically in combination therapies to prevent and treat malaria.

http://ift.tt/2mLDvBa

Population pharmacokinetics of finafloxacin in healthy volunteers and patients with complicated urinary tract infections [PublishAheadOfPrint]

Background: Finafloxacin is a novel fluoroquinolone with increased antibacterial activity at acidic pH and reduced susceptibility to several resistance mechanisms. A phase II study revealed a good efficacy/safety profile in patients with complicated urinary tract infections (cUTI) while pharmacokinetics was characterized by highly variable concentration vs. time profiles, suggesting the need for an elaborated pharmacokinetic model.

Methods: Data from three clinical trials were evaluated; 127 healthy volunteers were dosed orally (N=77) or intravenously (N=50) and 139 patients with cUTI received finafloxacin intravenously. Plasma (2824 samples in volunteers, 414 in patients) and urine (496 samples in volunteers, 135 in patients) concentrations were quantified by LC/MS-MS. NONMEM was used to build a population pharmacokinetic model and pharmacokinetic/pharmacodynamic relationships were investigated via simulations and logistic regression.

Results: A two-compartment model with first-order elimination described the data best (central [Vc] and peripheral [Vp] volumes: 47L [20%] and 43L [67%]; elimination clearance and inter-compartmental clearance: 21L/h [54%] and 2.8L/h [57%]) (median bootstrap estimates, coefficients of variation). Vc increased with body surface area and clearance was reduced in patients (-29%). Oral absorption was described best by parallel first- and zero-order processes (bioavailability 75%). No pharmacodynamic surrogate parameter of clinical/microbiological outcome could be identified, which depended exclusively on the MIC of the causative pathogens.

Conclusions: Despite the inter-individual variability, the present dataset does not support covariate-based dose adjustments. Based on the favorable safety and efficacy data, the clinical relevance of the observed variability appears to be limited.

http://ift.tt/2rhgevo

Efficacy and Safety of Finafloxacin versus Ciprofloxacin in the Treatment of Complicated Urinary Tract Infections: An Explorative Randomized Phase II Clinical Study. [PublishAheadOfPrint]

Introduction

The broad spectrum C-8-cyano-fluoroquinolone finafloxacin displays enhanced activity at acidic conditions. This phase II clinical study compared efficacy and safety of finafloxacin and ciprofloxacin in patients with complicated urinary tract infection and/or pyelonephritis.

Results

A 5-day regimen with finafloxacin 800 mg q.d. (FINA05) had similar results as a 10-day regimen with finafloxacin 800 mg q.d. (FINA10). Combined microbiological and clinical responses at test of cure visit (TOC) were 70% for FINA05, 68% for FINA10, and 57% for a 10-day ciprofloxacin regimen (CIPRO10) in 193 (64 for FINA05, 68 for FINA10 and 61 for CIPRO10) patients of the microbiological intent-to-treat (mITT) population. Additionally, the clinical effects of ciprofloxacin in patients with an acidic urine pH (80% of patients) were reduced whereas the effects of finafloxacin were unchanged. Finafloxacin was safe and well tolerated. Overall 43.4% of the patients in FINA05, 42.7% in FINA10 and 54.2% in the CIPRO10 group experienced mostly mild and treatment emergent, but unrelated adverse events.

Conclusion

A short course regimen of five days with finafloxacin resulted in higher eradication and improved clinical outcome rates than treatment with ciprofloxacin for 10 days. In contrast to ciprofloxacin, the clinical effects of finafloxacin were not reduced by acidic urine pH.

Methods

Hospitalized adults were randomized 1:1:1 to finafloxacin (800 mg q.d.) either for 5 or 10 days or ciprofloxacin (400 mg/500 mg b.i.d.) for 10 days with an optional switch from i.v. to oral administration at day 3. The primary endpoint was the combined microbiological and clinical response at TOC in the microbiological intent-to-treat population.

http://ift.tt/2mLL3UA

In vitro antibiotic susceptibility pattern of Non-diphtheriae Corynebacterium in Ontario, 2011-2016. [PublishAheadOfPrint]

Non-diphtheriae Corynebacterium-associated disease has been increasingly observed and often presents a conundrum to the treating physician. Analysis of antibiotic susceptibility test data for 1,970 clinical Corynebacterium isolates received between 2011 and 2016 revealed that empiric drug treatment options are limited to vancomycin and linezolid. Corynebacterium striatum was the most frequently observed species during this study period along with C. amycolatum and C. pseudodiphtheriticum/propinquum. Low levels of susceptibility to penicillin (14.5%), erythromycin (15.1%) and clindamycin (8.7%) were observed for non-diphtheriae Corynebacterium species, while 3.0% of isolates were not susceptible to daptomycin. Similarly, 26.9% and 38.1% of Corynebacterium isolates were susceptible to ciprofloxacin and trimethoprim-sulfamethoxazole, respectively. Our data shows much lower susceptibility to penicillin than previously reported in literature and an increasing number of isolates resistant to daptomycin highlighting the need for continued antibiotic surveillance studies for appropriate patient management and treatment success.

http://ift.tt/2rejHLw

Combating carbapenem-resistant Acinetobacter baumannii by an optimized imipenem plus tobramycin dosage regimen - prospective validation via hollow fiber infection and mathematical modeling [PublishAheadOfPrint]

We aimed to prospectively validate an optimized combination dosage regimen against a clinical carbapenem-resistant Acinetobacter baumannii (CRAB; MIC_Imipenem: 32mg/liter, MIC_Tobramycin: 2mg/liter). Imipenem at constant concentrations (7.6, 13.4 and 23.3mg/liter, reflecting a range of clearances) was simulated in a 7-day hollow fiber infection model (inoculum ~10^7.2 CFU/mL) with and without tobramycin (7mg/kg q24h, 0.5h infusions). While monotherapies achieved no killing or failed by 24h, this rationally optimized combination achieved >5 log₁₀ bacterial killing and suppressed resistance.

http://ift.tt/2mMayF9

Efficacy of Apramycin against Multidrug-Resistant Acinetobacter baumannii in the Murine Neutropenic Thigh Model [PublishAheadOfPrint]

Apramycin, an aminocyclitol aminoglycoside, was rapidly bactericidal against Acinetobacter baumannii. In a neutropenic murine thigh infection model, treatment-associated A. baumannii CFU reduction of greater than 4-log₁₀ per thigh was observed for all exposures for which AUC/MIC was greater than 50 and C_max/MIC was 10 or higher. Based on these findings, we suggest that apramycin deserves further pre-clinical exploration as a repurposed therapeutic for the treatment of multidrug-resistant, Gram-negative pathogens including A. baumannii.

http://ift.tt/2rk0BUn

Pterostilbene, a potential MCR-1 inhibitor that enhances the efficacy of polymyxin B [PublishAheadOfPrint]

We characterized pterostilbene produced a synergistic effect with polymyxin B (FIC index=0.156 or 0.188) against MCR-producing Escherichia coli strains of both human and animal origin. The time-killing assays showed that either pterostilbene or polymyxin B failed to eradicate a mcr-1- and NDM-positive E. coli strain ZJ487, but the combination eliminated by 1-hour post-inoculation. The survival rate of mouse following the intraperitoneal infection significantly promoted from 0% to 60% in the group of combination therapy applied.

http://ift.tt/2mNptyQ

The genetic basis of emerging vancomycin, linezolid, and daptomycin heteroresistance in a case of persistent Enterococcus faecium bacteremia [PublishAheadOfPrint]

Whole-genome sequencing was used to examine a persistent E. faecium bacteremia that acquired heteroresistance to three antibiotics in response to prolonged multidrug therapy. Comparison of complete genomes before and after each change revealed the emergence of known resistance determinants for vancomycin and linezolid, and suggested that a novel mutation in fabF, encoding a fatty acid synthase, was responsible for daptomycin nonsusceptibility. Plasmid recombination contributed to progressive loss of vancomycin resistance after withdrawal of the drug.

http://ift.tt/2rf7LZU

In vitro susceptibility of Neisseria gonorrhoeae strains to mupirocin. An antibiotic reformulated to parenteral nano-liposomal antibiotic [PublishAheadOfPrint]

Neisseria gonorrhoeae is an urgent antibiotic-resistant threat. This study determined the minimum inhibitory concentrations (MIC) of mupirocin to be 0.0039-0.0625 μg/ml for 94 N. gonorrhoeae strains. Cross-resistance with other antibiotics was not detected.

Mupirocin, currently limited to topical administration, demonstrated activity by injection when delivered in nano-liposomes. The nano-liposomal formulation of mupirocin is a potential treatment for drug-resistant N. gonorrhoeae.

http://ift.tt/2mNRvdx

New shuttle vectors for gene cloning and expression in multidrug-resistant Acinetobacter species [PublishAheadOfPrint]

Understanding bacterial pathogenesis requires adequate genetic tools to assess the role of individual virulence determinants by mutagenesis and complementation assays, as well as for homologous and heterologous expression of cloned genes. Our knowledge of Acinetobacter baumannii pathogenesis has so far been limited by the scarcity of genetic tools to manipulate multi drug resistant (MDR) epidemic strains which are responsible for most of infections. Here, we report the construction of new multi-purpose shuttle plasmids, namely pVRL1 and pVRL2, which can efficiently replicate in Acinetobacter sp. and in Escherichia coli. The pVRL1 plasmid has been constructed by combining: i) the cryptic plasmid pWH1277 from Acinetobacter calcoaceticus, which provides an origin of replication for Acinetobacter sp.; ii) a ColE1-like origin of replication; iii) the gentamicin or zeocin resistance cassette for antibiotic selection; iv) a multilinker containing several unique restriction sites. Modification of pVRL1 led to the generation of the pVRL2 plasmid, which allows arabinose-inducible gene transcription, with undetectable basal expression level of cloned genes under un-induced conditions and high-dynamic range of responsiveness to the inducer. Both pVRL1 and pVRL2 can easily be selected in MDR A. baumannii, have narrow host range and high copy number, are stably maintained in Acinetobacter sp., and appear compatible with indigenous plasmids carried by epidemic strains. Plasmid maintenance is guaranteed by the presence of a toxin-antitoxin system, providing more insights into the mechanism of plasmids stability in Acinetobacter sp.

http://ift.tt/2rf7LJo

A novel inhibitor of the LolCDE ABC transporter essential for lipoprotein trafficking in Gram-negative bacteria [PublishAheadOfPrint]

The outer membrane is an essential structural component of Gram-negative bacteria that is composed of lipoproteins, lipopolysaccharides, phospholipids, and integral β-barrel membrane proteins. A dedicated machinery, called the Lol system, ensures proper trafficking of lipoproteins from the inner to the outer membrane. The LolCDE ABC transporter is the inner membrane component, which is essential for bacterial viability. Here, we report a novel pyrrolopyrimidinedione compound, G0507, which was identified in a phenotypic screen for inhibitors of E. coli growth followed by selection of compounds that induced the extracytoplasmic ^E stress response. Mutations in lolC, lolD, and lolE conferred resistance to G0507 suggesting LolCDE as its molecular target. Treatment of E. coli cells with G0507 resulted in accumulation of fully processed Lpp, an outer membrane lipoprotein, in the inner membrane. Using purified protein complexes, we found that G0507 binds to LolCDE and stimulates its ATPase activity. G0507 still binds to LolCDE harboring a LolC_Q258K mutation, which confers high-level resistance to G0507 in vivo, but no longer stimulates ATPase activity. Our work demonstrates that G0507 has significant promise as chemical probe to dissect lipoprotein trafficking in Gram-negative bacteria.

http://ift.tt/2mM8YDe

Effectiveness of an Antimicrobial Stewardship Program in Critical Care: A Before-and-After Study [PublishAheadOfPrint]

We evaluated the use of antimicrobials expressed as defined daily doses (DDDs) per 1,000 patient days and days of therapy (DOT) per 100 occupied bed-days in a intensive care unit (UCI) of a general hospital in Barcelona, Spain, before and after implementation of an antimicrobial stewardship (AMS) program (2007-2010 versus 2011-2015). The quarterly costs of antimicrobials used in the ICU and its weight in the overall hospital costs of antimicrobials were calculated. The effect of the applied AMS program on DDDs and DOT time series data was analyzed by means of intervention time series analysis. A total of 5,002 patients were included (1,971 for the first [before] period and 3,031 for the second [after] period). The percentage of patients treated with one or more antimicrobials decreased from 88.6% to 77.2% (P< 0.001). DDDs decreased from 246.8 to 192.3 (mean difference -54.5, P = 0.001) and DOT from 66.7 to 54.6 (mean difference -12.1, P = 0.066). The mean cost per trimester decreased from 115,543 to 73,477 (mean difference -42,065.4, P< 0.001) and the percentage of ICU antimicrobials cost with respect to the total cost of hospital antimicrobials decreased from 28.5% to 22.8% (mean difference -5.59, P = 0.023). Implementation of an AMS program in the ICU was associated with a marked reduction of the use of antimicrobials, with cost savings close to one million euros since its implementation. An AMS program can have a significant impact on optimizing antimicrobial use in critical care practice.

http://ift.tt/2rdgYC3

Phosphonium Salt Displays Cytotoxic Effects Against Human Cancer Cell Lines

Aims/Objective: Phosphonium salts are compounds whose structural characteristics enable them to cross the plasma and mitochondrial membrane with ease. Cancer cells have higher plasma membrane potentials than normal cells; phosphonium salts selectively accumulate in the mitochondria of neoplastic cells and inhibit mitochondrial function.

Method: In the present work, we investigated the cytotoxic activity of lipophilic phosphonium salt (11- methoxy11-oxo-undecyl) triphenylphosphonium bromide (MUTP) as well as of the two new phosphine oxide salts, 3,3'-(methylphosphoryl) dibenzenaminium chloride (SBAMPO) and 3,3' (phenylphosphoryl) dibenzenaminium chloride (SBAPPO) on the proliferation of breast cancer cell line (MCF-7) and human uterin cervix adenocarcinoma cells (HeLa).

Result: We showed that only MUTP exhibits antiproliferative effects on both cell lines, without affecting the normal breast epithelial cell proliferation. More specifically, we demonstrated that MUTP treatment of breast cancer cells is associated with impaired cell-cycle progression and metabolically induces mitochondrial damage and triggers apoptotic cell death in MCF-7 and HeLa cells. Taken together, these findings suggest that MUTP may be capable of selectively targeting neoplastic cell growth and therefore has potential applications as anticancer agent.

http://ift.tt/2FNiLkw

Body Fluids-Derived Exosomes: Paving the Novel Road to Lung Cancer Diagnosis and Therapy

Lung cancer is a major human malignancy. Nowadays, the lack of specific diagnostic markers of lung cancer restricts the early diagnosis and therapy of patients. Exosomes, as spherical 30-100 nm microvesicles, are released by normal and cancer cells in both physiological and pathological circumstances. Exosomes carry various molecular cargos such as miRNA, proteins, mRNA, DNA and lipids. Therefore, analysis of the molecular profiles of exosomes may provide beneficial biomarkers for disease diagnosis. Exosomes can be transported by body fluids. The molecules (miRNAs and proteins) detected in body fluid exosomes may contribute to lung cancer diagnosis. In this review, we summarize typical molecules (miRNAs and proteins) in body fluids-derived exosomes to reveal the potential biomarkers in lung cancer. Besides, the role and the application of exosomes in chemotherapy and radiotherapy of lung cancer patients have also been discussed in this review.

http://ift.tt/2EOgJz8

Synthesis of Novel Benzothiazole-Piperazine Derivatives and Their Biological Evaluation as Acetylcholinesterase Inhibitors and Cytotoxic Agents

Objective and Method: A new series of benzothiazole-piperazine derivatives was synthesized and a complete chemical characterization of the novel compounds was provided. In vitro cytotoxic activities were screened against colorectal (HCT-116), breast (MCF-7) and hepatocellular (Huh7) cancer cell lines by Sulforhodamine B assay.

Result and Discussion: All compounds showed cytotoxic activity against hepatocellular (Huh7) and breast (MCF-7) cancer cell lines. Dihalo substituted benzylpiperazine derivatives (2a, 2e) had the highest cytotoxic activities in all the tested cell lines. In addition, acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibitory activities of synthesized compounds were investigated by in vitro Ellman's method. Compound 2j led to moderate and selective inhibition against AChE. Docking study was utilized to understand the binding mode of compound 2j in comparision with donepezil on AChE. The other tested compounds showed weak or no inhibition against AChE as promising anticancer agents.

http://ift.tt/2FJYhZF

Novel N-mustard-benzimidazoles/benzothiazoles Hybrids, Synthesis and Anticancer Evaluation

Background: Bendamustine, an N-mustard-benzoimidazole hybrid conjugate, was recently approved for the treatment of chronic lymphocytic leukemia. However, the short half-life of bendamustine may limit its clinical applications.

Objective: The purpose of this study is to design and synthesize compounds with a more favorable pharmacokinetic profile.

Methods: We synthesized a series of hybrid molecules comprising a phenyl N-mustard moiety and benzothiazole or benzimidazole scaffold linked via a urea linker and evaluated their antitumor activity and plasma stability.

Results: We revealed that these agents exhibited significant cytotoxicity against a panel of human lymphoblastic leukemia and human solid tumor cells in culture. Human lymphoblastic leukemia CCRM-CEM cells were the most sensitive to the tested compounds. In general, the new hybrids were as potent as cisplatin, but significantly more cytotoxic than bendamustine. Phenyl N-mustard-benzothiazole compound 27d and phenyl N-mustardbenzimidaloe compound 32b possessed significant cytotoxicity and led to apoptotic death in the treated tumor cells. These two agents were able to induce DNA interstrand cross-linking and arrested cell cycle progression at the G2/M phase. Furthermore, we showed that these new hybrids were more chemically stable than bendamustine in rat plasma.

Conclusion: Our results suggest that conjugation of phenyl N-mustard pharmacophore at C6 of benzimidazole or at C8 of the benzothiazole ring via a urea linker is likely an approach to increase the chemical stability and bioavailability.

Highlights ⇒ Series of benzimidazoles and benzothiazoles linked to N-mustard were synthesized. ⇒ The newly synthesized derivatives induced DNA interstrand cross-links. ⇒ These derivatives induced cell cycle arrest in the G2/M phase and triggered apoptosis in H460 cells. ⇒ The new compounds are more cytotoxic than bendamustine. ⇒ The new compounds were chemically more stable than bendamustine in rat plasma.

http://ift.tt/2ELB0W3

The Oxime Derivatives of 1-R-1H-Naphtho[2,3-d][1,2,3]triazole-4,9-dione 2-oxides: Synthesis and Properties

Objective: To synthesize a novel chemotype based on the naphthoquinone scaffold with retained cytotoxicity and provisionally low intracellular oxidation potential.

Background: Derivatives of naphthoquinone, although potent anticancer agents, can exert heart toxicity due to generation of free oxygen species.

Methods: In this study, we modified the scaffold by replacing one carbonyl group with the oxime moiety. Interestingly, only one carbonyl group in 1-R-1H-naphtho[2,3-d][1,2,3]triazole-4,9-dione 2-oxides reacted with hydroxylamine. The spatial structure was determined by X-ray analysis. New compounds were tested for the ability to form stable complexes with double stranded DNA by spectroscopy and molecular docking and to induce death of tumor cell lines and non-malignant counterparts.

Results: The resulting 1-R-1H-naphtho[2,3-d][1,2,3]triazole-4,9-dione 4-oxime 2-oxides were further acylated to produce a series of 1-R-1H-naphtho[2,3-d][1,2,3]triazole-4,9-dione 4-(O-acyloxime) 2-oxides. Newly synthesized compounds demonstrated a higher (in submicromolar or low micromolar range) cytotoxic potency against human colon and breast adenocarcinoma cell lines than to non-malignant skin fibroblasts. Spectroscopic measurements revealed that, unlike other classes of quinone derivatives, new naphthotriazoledione oxides did not form stable complexes with double stranded DNA regardless of their fitting to the DNA minor groove (as determined by molecular modeling).

Conclusion: Thus, our chemical modifications yielded a new chemotype with good cytotoxic properties and yet-to-be-identified intracellular target(s).

http://ift.tt/2FIZdO7

Design, Synthesis and Biological Evaluation of Novel 1, 3- thiazolidine-2, 4-diones as Anti-prostate Cancer Agents

Background: Androgen receptor is an attractive target for the treatment of prostate cancer. The 1,3- thiazolidine-2,4-diones possess a wide diversity of important biochemical effects and interesting pharmacological properties.

Objective: The aim of the study is to find the experimental and computational methods to investigate the interference of 1,3-thiazolidine-2,4-diones with androgen receptor against prostate cancer.

Method: Structural modification and molecular docking-based virtual screening approaches were imposed to identify the novel 1,3-thiazolidine-2,4-diones by using Schrödinger (Maestro 9.5). The best fit molecules (3-12 & 23-31) were synthesized and characterized using spectroscopic techniques, then in vitro antioxidant and antiprostate cancer activities were evaluated. Further, the structure of the intermediate (18) was confirmed by single crystal XRD analysis. The mechanism studies were performed through the gene expression for the compounds, 29, 30, and 31, the standards, dihydrotestosterone and R-bicalutamide.

Results: The compounds, 29, 30 and 31 showed comparatively significant antioxidant activity and better antiproliferative activity against PC-3 and LNCaP cell lines. Also, very low cytotoxicity was observed in the noncancerous cell (3T3). The compounds, 29, 30 and 31 significantly decreased the mRNA expression of ARstimulated genes, PSA and TMPRSS2, which demonstrated their anti-prostate cancer activities. ADME/T properties prediction of the compounds (3-12 and23-31) showed the promising drug-likeness and pharmacokinetic parameters without toxicity. Moreover, DFT calculations apparently confirmed the stable conformer of the compound, 31.

Conclusion: These findings may provide the essential information for the development of anti-prostate cancer agents.

http://ift.tt/2EQ16Hz

Meet Our Editorial Board Member

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Synthesis, Radiosynthesis and Metabolism of 131I-Y-c(CGRRAGGSC)

Background: The formation of the complex interleukin-11(IL-11) and IL-11 receptor (IL-11R) is closely related with tumor progression. Binding of IL-11 to the IL-11 receptor α-chain (IL-11Rα) has been suggested as a target for human cancer. The cyclic peptide c(CGRRAGGSC) is a mimic of IL-11.

Objective: To explore 131I-Y-c(CGRRAGGSC) synthesis and radiosynthesis, and metabolism in SKOV3 tumorbearing mice.

Method: In this study, 131I labeled c(CGRRAGGSC) was designed and characterized. For radiolabeling, tyrosine was used as a linker to connect c(CGRRAGGSC) and 131I. Balb/c nude mice bearing SKOV3 human ovarian carcinoma were used for in vivo studies. Uptake of 131I-cyclic nonapeptide by the tumor was visualized by single photon emission computerized tomography (SPECT).

Results: The entire labeling process, which took 15 min by chloramine-T method, resulted in a high labeling yield (93.03±6.78%), and high radiochemical purity (RCP) (>95%). SPECT imaging showed that accumulation of the probe in the tumor was close to background levels. In addition, biodistribution studies showed that the accumulation of 131I-Y-c(CGRRAGGSC) in normal mice was similar to that of Na131I.

Conclusion: Tyrosine is a suitable chelating agent for the use of radioiodine labeling, however the bioactivity of the conjugate needs further investigation.

http://ift.tt/2ELAXJR

Pharmacogenetics of Aromatase Inhibitors in Endocrine Responsive Breast Cancer: Lessons Learnt from Tamoxifen and CYP2D6 Genotyping

Background: Genetics play a significant role in drug metabolism of endocrine therapy of breast cancer. These aspects have been studied extensively in patients on tamoxifen, but the pharmacogenetics of aromatase inhibitors are less established. In contrast to the protective effect of tamoxifen, aromatase inhibitors are linked with an increased risk for bone loss and fractures.

Objective: This review outlines key issues in the implementation of pharmacogenetics of cytochrome P450 and tamoxifen as a model for optimal use of aromatase inhibitors in postmenopausal women with estrogen receptor positive breast cancer.

Methods: Lessons learnt from the association between tamoxifen and CYP2D6 genotyping were applied to identify polymorphisms with the potential to change clinical decision-making in patients on aromatase inhibitors. The ability of next generation sequencing to supersede single-gene analysis was furthermore evaluated in a subset of breast cancer patients on aromatase inhibitors selected from a central genomics database.

Results: Methodological flaws in major randomised controlled trials and continued referral to incorrect results in expert consensus statements are important factors delaying the implementation of CYP2D6 pharmacogenetics in tamoxifen treatment. This highlighted the importance of a clinical pipeline including comprehensive genotyping, to define the target population most likely to benefit from aromatase inhibitor pharmacogenetics.

Conclusion: The clinical utility of CYP2D6 genotyping is well-established in patients at increased risk of tamoxifen resistance due to cumulative risk. The pharmacogenetics of CYP19A1 requires further clarification in terms of bone risk assessment for appropriate use in the treatment algorithm of high-risk patients at the onset of aromatase inhibitors.

http://ift.tt/2FIJTAW

Design, Synthesis and Biological Evaluation of Betulinic Acid Derivatives as New Antitumor Agents for Leukemia

Background: Chronic myeloid leukemia (CML) is currently treated with imatinib, a Bcr-Abl inhibitor. However, resistance to this drug usually develops over time. Triptolide, a diterpenoid triepoxide, has been shown active against CML cells resistant to imatinib, acting mainly on the level of Bcr-Abl transcription inhibition.

Objective: Here, we used the triterpene betulinic acid, a known proteasome inhibitor with potential antileukemic activity, as a scaffold for the generation of analogues with predicted triptolide biological activity.

Method: Betulinic acid derivatives were designed based on the structure-activity relationship of triptolide and evaluated for their cytotoxic effects in CML cells, lymphocytes and human keratinocytes (HaCaT), as well as against the proteasome complex. The main modification performed on betulinic acid was fluorination at C-28 and epoxidation, both of which are responsible for enhancing activity of triptolide. A total of 10 compounds were obtained: 6 previously described and 4 novel compounds. The cytotoxic activity over a CML cell line (K562) was assessed using flow cytometry and compared to lymphocytes and HaCaT.

Result: The results show that betulinic acid was the most cytotoxic compound against CML cells, showing a good selectivity index for cancer over normal cells. The most important trend for the activity in betulinic acid derivatives is the presence of a free hydroxyl group at C-3 and a carboxyl group at C-28. Results also indicated that the epoxide is important for enhancing the activity, while modification at C-28 worsens the activity.

Conclusion: Proteasome inhibition assays suggest that proteasome is the main target for betulinic acid and its derivatives.

http://ift.tt/2ENBMBM

Enhancement of the Effect of Methyl Pyropheophorbide-a-Mediated Photodynamic Therapy was Achieved by Increasing ROS through Inhibition of Nrf2-HO-1 or Nrf2-ABCG2 Signaling

Background: Emerging evidence indicates that the transcription factor nuclear factor-E2-related factor 2 (Nrf2) plays an essential role in cellular defense against oxidative stress; its activation has been related to cytoprotection.

Objective: Here, we investigated the role of Nrf2 in improving the efficacy of methyl pyropheophorbide-amediated photodynamic therapy (Mppa-PDT) via the downregulation of Nrf2.

Method: Human ovarian cancer A2780 cells and SKOV3 cells were treated with Mppa-PDT and siRNA transfection was performed to inhibit Nrf2. After treated with siRNA and Mppa-PDT, the cell viability was examined with CCK-8 assay; cell apoptosis was detected tested by flow cytometry with Annexin V-FITC/PI; the celluar reactive oxygen species (ROS) and mitochondrial membrane potential were measured with DCFHDA and JC-1 staining; expression of protein was assessed by western blot analysis.

Results: We found that Nrf2 translocated from the cytoplasm to the nucleus in vitro and in vivo, and the expression of Nrf2 and P-Nrf2 increased through a possible mechanism regulated by mitogen-activated protein kinase (MAPK) after Mppa-PDT treatment. Furthermore, cytotoxicity and apoptosis induced by Mppa-PDT increased after Nrf2down-regulation. Nrf2 down -regulation increased reactive oxygen species (ROS) levels by attenuating antioxidants or pumping Mppa out of cells,which resulted from the inhibition of Nrf2-HO-1 or Nrf2- ABCG2 signaling. In addition, SKOV3 cells exhibited increased resistance to Mppa-PDT, and the expression levels of P-Nrf2 and ABCG2 were higher in SKOV3 cells than in A2780 cells, suggesting that Nrf2-ABCG2 signaling might be involved in the intrinsic resistanceto Mppa-PDT.

Conclusion: These results provided evidence that Nrf2 down-regulation can enhance the effect of Mppa-PDT.

http://ift.tt/2FK8FAX

Myc Decoy Oligodeoxynucleotide Inhibits Growth and Modulates Differentiation of Mouse Embryonic Stem Cells as a Model of Cancer Stem Cells

Background: Myc (c-Myc) alone activates the embryonic stem cell-like transcriptional module in both normal and transformed cells. Its dysregulation might lead to increased cancer stem cells (CSCs) population in some tumor cells.

Objective: In order to investigate the potential of Myc decoy oligodeoxynucleotides for differentiation therapy, mouse embryonic stem cells (mESCs) were used in this study as a model of CSCs. To our best of knowledge this is the first report outlining the application of Myc decoy in transcription factor decoy "TFD" strategy for inducing differentiation in mESCs.

Methods: A 20-mer double-stranded Myc transcription factor decoy and scrambled oligodeoxynucleotides (ODNs) were designed, analyzed by electrophoretic mobility shift (EMSA) assay and transfected into the mESCs under 2 inhibitors (2i) condition. Further investigations were carried out using fluorescence and confocal microscopy, cell proliferation and apoptosis analysis, alkaline phosphatase and embryoid body formation assay, real-time PCR and western blotting.

Results: EMSA data showed that Myc decoy ODNs bound specifically to c-Myc protein. They were found to be localized in both cytoplasm and nucleus of mESCs. Our results revealed the potential capability of Myc decoy ODNs to decrease cell viability by (16.1±2%), to increase the number of cells arrested in G0/G1 phases and apoptosis by (14.2±3.1%) and (12.1±3.2%), respectively regarding the controls. Myc decoy could also modulate differentiation in mESCs despite the presence of 2i/LIF in our medium the presence of 2i/LIF in our medium.

Conclusion: The optimized Myc decoy ODNs approach might be considered as a promising alternative strategy for differentiation therapy investigations.

http://ift.tt/2EM4tPu

Biological Activity Evaluation of Novel 1,2,4-Triazine Derivatives Containing Thiazole/Benzothiazole Rings

Background: Triazine ring is a prominent structural motif found in some azanucleosides whose efficiency improved many times in the research area of antitumor agents.

Objective: In this study, we have designed and synthesized novel 2-[(5,6-diphenyl-1,2,4-triazin-3-yl)thio]-N-(6- substituted benzo/(thiazol)-2-yl)acetamide (2a-d, 3a-f) derivatives using 1,2,4-triazine core along with two important heterocyles, thiazole and benzothiazole rings.

Method: The acquired ten final compounds were screened to investigate their antitumor activity against lung adenocarcinoma cell line, A549 and mouse fibroblast cell line, NIH/3T3. Five compounds with higher antiproliferative activity have been further studied to evaluate whether the cell death due to necrosis or apoptosis using flow cytometry.

Results and Conclusion: Compound 3b bearing 6-methylbenzothiazole moiety has been established as the most active antitumor compound with a selective profile and higher apoptotic cell level. All final componds were also screened against acetylcholine/butyrylcholinesterase enzymes to state their anticholinesterase activity.

http://ift.tt/2FLkXZs

First Comprehensive Companion Diagnostic OK'd [News in Brief]

Test uses next-generation sequencing to determine mutation status for 324 genes.

http://ift.tt/2FKITMA

Genes predisposed to DNA hypermethylation during acquired resistance to chemotherapy are identified in ovarian tumors by bivalent chromatin domains at initial diagnosis

Bivalent chromatin domains containing both active H3K4me3 and repressive H3K27me3 histone marks define gene sets poised for expression or silencing in differentiating embryonic stem (ES) cells. In cancer cells aberrantly poised genes may facilitate changes in transcriptional states after exposure to anti-cancer drugs. In this study, we used ChIP-seq to characterize genome-wide positioning of H3K4me3 and H3K27me3-associated chromatin in primary high-grade serous ovarian carcinomas and in normal ovarian surface and fallopian tube tissue. Gene sets with proximal bivalent marks defined in this manner were evaluated subsequently as signatures of systematic change in DNA methylation and gene expression, comparing pairs of tissue samples taken from patients at primary presentation and relapse following chemotherapy. We found that gene sets harboring bivalent chromatin domains at their promoters in tumor tissue, but not normal epithelia, overlapped with Polycomb-repressive complex target genes as well as transcriptionally silenced genes in normal ovarian and tubal stem cells. The bivalently marked genes we identified in tumors before chemotherapy displayed increased promoter CpG methylation and reduced gene expression at relapse after chemotherapy of ovarian cancer. Overall, our results support the hypothesis that pre-existing histone modifications at genes in a poised chromatin state may lead to epigenetic silencing during acquired drug resistance.

http://ift.tt/2Dhd2pb

Role of chromatin damage and chromatin trapping of FACT in mediating the anticancer cytotoxicity of DNA-binding small molecule drugs

Precisely how DNA-targeting chemotherapeutic drugs trigger cancer cell death remains unclear, as it is difficult to separate direct DNA damage from chromatin damage in cells. Recent work on curaxins, a class of small molecule drugs with broad anticancer activity, show that they interfere with histone-DNA interactions and destabilize nucleosomes without binding DNA or causing detectable DNA damage. Chromatin unfolding caused by curaxins is sensed by the histone chaperone FACT, which binds unfolded nucleosomes and causes chromatin trapping (c-trapping). In this study, we investigated whether classical DNA-targeting chemotherapeutic drugs also similarly disturbed chromatin to cause c-trapping. Drugs that directly bound DNA induced both chromatin damage and c-trapping. However, chromatin damage occurred in the absence of direct DNA damage and was dependent on how a drug bound DNA, specifically, in the way it bound chromatinized DNA in cells. FACT was sensitive to a plethora of nucleosome perturbations induced by DNA-binding small molecules, including displacement of the linker histone, eviction of core histones, and accumulation of negative supercoiling. Strikingly, we found that the cytotoxicity of DNA-binding small molecules correlated with their ability to cause chromatin damage, not DNA damage. Our results suggest implications for the development of chromatin-damaging agents as selective anticancer drugs.

http://ift.tt/2B75BuY

AMPK-AKT double negative feedback loop in breast cancer cells regulates their adaptation to matrix deprivation

Cell detachment from the extracellular matrix triggers anoikis. Disseminated tumor cells must adapt to survive matrix deprivation, while still retaining the ability to attach at secondary sites and re-initiate cell division. In this study, we illuminate mechanisms that enable reversible matrix attachment by breast cancer cells. Matrix deprival triggered AMPK activity and concomitantly inhibited AKT activity by upregulating the AKT phosphatase PHLPP2. The resultant pAMPKhigh/pAKTlow state was critical for cell survival in suspension, as PHLPP2 silencing also increased anoikis while impairing autophagy and metastasis. In contrast, matrix re-attachment led to AKT-mediated AMPK inactivation via PP2C-α-mediated restoration of the pAKThigh/pAMPKlow state. Clinical specimens of primary and metastatic breast cancer displayed an AKT-associated gene expression signature, whereas circulating breast tumor cells displayed an elevated AMPK-dependent gene expression signature. Our work establishes a double-negative feedback loop between AKT and AMPK to control the switch between matrix-attached and matrix-detached states needed to coordinate cell growth and survival during metastasis.

http://ift.tt/2DhFirP

PHD3 controls lung cancer metastasis and resistance to EGFR inhibitors through TGF{alpha}

Lung cancer is the leading cause of cancer-related death worldwide, in large part due to its high propensity to metastasize and develop therapy resistance. Adaptive responses to hypoxia and epithelial-mesenchymal transition (EMT) are linked to tumor metastasis and drug resistance, but little is known about how oxygen sensing and EMT intersect to control these hallmarks of cancer. Here we show that the oxygen sensor PHD3 links hypoxic signaling and EMT regulation in the lung tumor microenvironment. PHD3 was repressed by signals that induce EMT and acted as a negative regulator of EMT, metastasis and therapeutic resistance. PHD3 depletion in tumors, which can be caused by the EMT inducer TGFβ or by promoter methylation, enhanced EMT and spontaneous metastasis via HIF-dependent upregulation of the EGFR ligand TGFα. In turn, TGFα stimulated EGFR which potentiated SMAD signaling, reinforcing EMT and metastasis. In clinical specimens of lung cancer, reduced PHD3 expression was linked to poor prognosis and to therapeutic resistance against EGFR inhibitors such as erlotinib. Re-expression of PHD3 in lung cancer cells suppressed EMT and metastasis and restored sensitivity to erlotinib. Taken together, our results establish a key function for PHD3 in metastasis and drug resistance and suggest opportunities to improve patient treatment by interfering with the feedforward signaling mechanisms activated by PHD3 silencing.

http://ift.tt/2B5PkXk

E6 protein expressed by high-risk HPV activates super-enhancers of the EGFR and c-MET oncogenes by destabilizing the histone demethylase KDM5C

The high-risk (HR) human papillomaviruses (HPV) are causative agents of anogenital tract dysplasia and cancers and a fraction of head and neck cancers. The HR HPV E6 oncoprotein possesses canonical oncogenic functions, such as p53 degradation and telomerase activation. It is also capable of stimulating expression of several oncogenes, but the molecular mechanism underlying these events are poorly understood. Here we provide evidence that HPV16 E6 physically interacts with histone H3K4 demethylase KDM5C, resulting in its degradation in an E3 ligase E6AP- and proteasome-dependent manner. Moreover, we found that HPV16-positive cancer cell lines exhibited lower KDM5C protein levels than HPV-negative cancer cell lines. Restoration of KDM5C significantly suppressed the tumorigenicity of CaSki cells, an HPV16-positive cervical cancer cell line. Whole genome ChIP-seq and RNA-seq results revealed that CaSki cells contained super-enhancers in the proto-oncogenes EGFR and c-MET. Ectopic KDM5C dampened these super-enhancers and reduced the expression of proto-oncogenes. This effect was likely mediated by modulating H3K4me3/H3K4me1 dynamics and decreasing bi-directional eRNA transcription. Depletion of KDM5C or HPV-16 E6 expression activated these two super-enhancers. These results illuminate a pivotal relationship between the oncogenic E6 proteins expressed by high-risk HPV isotypes and epigenetic activation of super-enhancers in the genome that drive expression of key oncogenes like EGFR and c-MET.

http://ift.tt/2Dk076g

Radio-resistant Cervical Cancers Are Sensitive to Inhibition of Glycolysis and Redox Metabolism

Highly glycolytic cervical cancers largely resist treatment by cisplatin and co-administered pelvic irradiation as the present standard of care. In this study, we investigated the effects of inhibiting glycolysis and thiol redox metabolism to evaluate them as alternate treatment strategies in these cancers. In a panel of multiple cervical cancer cell lines, we evaluated sensitivity to inhibition of glycolysis (2-DG) with or without simultaneous inhibition of glutathione and thioredoxin metabolism (BSO/AUR). Intracellular levels of total and oxidized glutathione, thioredoxin reductase activity, and indirect measures of intracellular reactive oxygen species (ROS) were compared before and after treatment. Highly radio-resistant cells were the most sensitive to 2-DG, whereas intermediate radio-resistant cells were sensitive to 2-DG plus BSO/AUR. In response to 2-DG/BSO/AUR treatment, we observed increased levels of intracellular oxidized glutatione, redox-sensitive dye oxidation and decreased glucose utilization via multiple metabolic pathways including the TCA cycle. 2-DG/BSO/AUR treatment delayed the growth of tumors composed of intermediate radio-resistant cells and effectively radio-sensitized these tumors at clinically relevant radiation doses both in vitro and in vivo. Overall, our results support inhibition of glycolysis and intracellular redox metabolism as an effective alternative drug strategy for the treatment of highly glycolytic and radio-resistant cervical cancers.

http://ift.tt/2B75gII

The Hippo pathway component TAZ promotes immune evasion in human cancer through PD-L1

The Hippo pathway component WW domain-containing transcription regulator 1 (TAZ) is a transcriptional co-activator and an oncogene in breast and lung cancer. Transcriptional targets of TAZ that modulate immune cell function in the tumour microenvironment are poorly understood. Here, we perform a comprehensive screen for immune-related genes regulated by TAZ and its paralog YAP using NanoString gene expression profiling. We identify the immune checkpoint molecule PD-L1 as a target of Hippo signaling. The upstream kinases of the Hippo pathway, mammalian STE20-like kinase 1 and 2 (MST1/2) and large tumour suppressor 1 and 2 (LATS1/2), suppress PD-L1 expression while TAZ and YAP enhance PD-L1 levels in breast and lung cancer cell lines. PD-L1 expression in cancer cell lines is determined by TAZ activity and TAZ/YAP/TEAD increase PD-L1 promoter activity. Critically, TAZ-induced PD-L1 upregulation in human cancer cells is sufficient to inhibit T cell function. The relationship between TAZ and PD-L1 is not conserved in multiple mouse cell lines, likely due to differences between the human and mouse PD-L1 promoters. To explore the extent of divergence in TAZ immune-related targets between human and mouse cells, we perform a second NanoString screen using mouse cell lines. We show that many targets of TAZ may be differentially regulated between these species. These findings highlight the role of Hippo signaling in modifying human/murine physiological/pathological immune responses and provide evidence implicating TAZ in human cancer immune evasion.

http://ift.tt/2DjZY2I

STAT3/PIAS3 levels serve as "early signature" genes in the development of high-grade serous carcinoma from the fallopian tube

The initial molecular events that lead to malignant transformation of the fimbria of the fallopian tube (FT) through high-grade serous ovarian carcinoma (HGSC) remain poorly understood. In this study, we report that increased expression of signal transducer and activator of transcription 3 (pSTAT3 Tyr705) and suppression or loss of protein inhibitor of activated STAT3 (PIAS3) in FT likely drive HGSC. We evaluated human tissues-benign normal FT, tubal peritoneal junction (TPJ), p53 signature fallopian tube tissue, tubal intraepithelial lesion in transition (TILT), serous tubal intraepithelial carcinoma (STIC) without ovarian cancer, and HGSC for expression of STAT3/PIAS3 (compared with their known TP53 signature) and their target proliferation genes. We observed constitutive activation of STAT3 and low levels or loss of PIAS3 in the TPJ, p53 signature, TILT, and STIC through advanced stage IV (HGSC) tissues. Elevated expression of pSTAT3 Tyr705 and decreased levels of PIAS3 appeared as early as TPJ and the trend continued until very advanced stage HGSC (compared to high PIAS3 and low pSTAT3 expression in normal benign FT). Exogenous expression of STAT3 in FT cells mediated translocation of pSTAT3 and c-Myc into the nucleus. In vivo experiments demonstrated that overexpression of STAT3 in fallopian tube secretory epithelial cells (FTSEC) promoted tumor progression and metastasis, mimicking the clinical disease observed in patients with HGSC. Thus we conclude that the STAT3 pathway plays a role in the development and progression of HGSC from its earliest pre-malignant states.

http://ift.tt/2B7h51w

Herbal formula YGJDSJ inhibits anchorage-independent growth and induces anoikis in hepatocellular carcinoma Bel-7402 cells

Based on clinical medications and related studies, we established a Yang-Gan Jie-Du Sang-Jie (YGJDSJ) herbal formula for hepatocarcinoma treatment. In present study, we evaluated the anti-cancer potential of Y...

http://ift.tt/2DE3TnR

Development and validation of HPTLC fingerprints of three species of Alpinia with biomarker Galangin

Alpinia galanga (L.) Willd. commonly called as Rasna, Greater galangal or Kulinjan is a medicinally important rhizome used in Indian traditional system of medicine to cure a number of ailments. A. galanga is the ...

http://ift.tt/2Daweku

Reduced Growth Hormone Secretion is Associated with Nonalcoholic Fatty Liver Disease in Obese Children

Horm Metab Res
DOI: 10.1055/s-0043-124970

The purpose of the study was to evaluate the relationship between arginine-levodopa-induced growth hormone (GH) secretion and nonalcoholic fatty liver disease (NAFLD) in obese children. This study includes a total of 84 obese and 43 normal weight children. The obese subjects are divided into two groups based on the presence or absence of NAFLD. Clinical examination, anthropometric and laboratory examinations, and liver ultrasonography are assessed for all participants. The obese group had significantly lower peak stimulated GH (p<0.001) and lower insulin-like growth factor 1 (IGF-1) (p<0.001) compared with the control group. Children with NAFLD had significantly lower peak stimulated GH (p<0.001) and lower IGF-1 (p=0.022) compared with non-NAFLD group. Results from logistic regression model showed that only peak GH after stimulation test was inversely associated with NAFLD (p=0.015), while body mass index (BMI) was positively associated with NAFLD (p=0.03). Among 84 obese children and adolescents, peak stimulated GH was negatively associated with alanine aminotransferase (r=–0.394, p<0.001), BMI (r=–0.571, p<0.001), systolic blood pressure (r=–0.223, p=0.041), diastolic blood pressure (r=–0.272, p=0.012), homeostasis model assessment of insulin resistance (r=–0.369, p=0.001), insulin (r=–0.382, p<0.001), and positively associated with high density lipoprotein cholesterol (r=0.275, p=0.011). Our study confirms a significant inverse relationship between NAFLD and GH response to standard stimulation testing in obese children without known hypothalamic/pituitary disease.
[...]

© Georg Thieme Verlag KG Stuttgart · New York

Article in Thieme eJournals:
Table of contents  |  Abstract  |  Full text

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Ecotropic viral integration site 1 promotes metastasis independent of epithelial mesenchymal transition in colon cancer cells

Ecotropic viral integration site 1 promotes metastasis independent of epithelial mesenchymal transition in colon cancer cells

Ecotropic viral integration site 1 promotes metastasis independent of epithelial mesenchymal transition in colon cancer cells, Published online: 16 January 2018; doi:10.1038/s41419-017-0036-1

Ecotropic viral integration site 1 promotes metastasis independent of epithelial mesenchymal transition in colon cancer cells

http://ift.tt/2mFmDuP

How BMP-2 induces EMT and breast cancer stemness through Rb and CD44?

How BMP-2 induces EMT and breast cancer stemness through Rb and CD44?

How BMP-2 induces EMT and breast cancer stemness through Rb and CD44?, Published online: 16 January 2018; doi:10.1038/s41419-017-0037-0

How BMP-2 induces EMT and breast cancer stemness through Rb and CD44?

http://ift.tt/2DoJGVc

MCL-1 is a prognostic indicator and drug target in breast cancer

MCL-1 is a prognostic indicator and drug target in breast cancer

MCL-1 is a prognostic indicator and drug target in breast cancer, Published online: 16 January 2018; doi:10.1038/s41419-017-0035-2

MCL-1 is a prognostic indicator and drug target in breast cancer

http://ift.tt/2mDyOIA

Plumbagin inhibits the proliferation and survival of esophageal cancer cells by blocking STAT3-PLK1-AKT signaling

Plumbagin inhibits the proliferation and survival of esophageal cancer cells by blocking STAT3-PLK1-AKT signaling

Plumbagin inhibits the proliferation and survival of esophageal cancer cells by blocking STAT3-PLK1-AKT signaling, Published online: 16 January 2018; doi:10.1038/s41419-017-0068-6

Plumbagin inhibits the proliferation and survival of esophageal cancer cells by blocking STAT3-PLK1-AKT signaling

http://ift.tt/2Dpe4Pc

Dexamethasone alleviates allergic asthma immature rat through Toll like receptor 4

OBJECTIVE: The allergic asthma model induced by ovalbumin (OVA) was established in the immature rat. Dexamethasone (DXM) was adopted for intervention to analyze the treatment effect and to explore the relationship with toll-like receptor 4 (TLR4).

MATERIALS AND METHODS: Immature SD rat was treated by OVA to construct allergic asthma model and intervened by DXM. The rats were randomly divided into model group, experimental group, and control group. The changes in lung tissue were observed by light microscope. The EOS infiltration and reactivity of airway wall were compared. The expressions of TLR2 and TLR4 protein and mRNA in the lung tissue were tested by Western blot and RT-PCR.

RESULTS: The lung tissue in the model group was infiltrated by a lot of inflammatory cells, and mucous membrane edema was observed, compared with that in the control group. There were only a few inflammatory cells in the interstitial tissue and pulmonary alveoli in the experimental group compared with that in the model group. EOS count of airway wall and airway reactivity decreased in the experimental group. The levels of TLR2 and TLR4 were significantly elevated in the third week compared with the first week (p<0.05).

CONCLUSIONS: The treatment of DXM can alleviate the pathological changes of the lung tissue in SD immature rat with allergic asthma, reduce EOS infiltration in the airway wall, decrease airway reactivity, and elevate expressions of TLR2 and TLR4.

L'articolo Dexamethasone alleviates allergic asthma immature rat through Toll like receptor 4 sembra essere il primo su European Review.

http://ift.tt/2DEn0hq

Prophylactic use of non-invasive mechanical ventilation in lung resection

OBJECTIVE: To evaluate if the prophylactic application of BiPAP previous to lung resection and 17 hours postoperatively improves respiratory function. In order to do this, we studied the results of arterial blood gases and portable spirometry in the immediate postoperative period and at the first and third postoperative day. Secondary objectives included evaluating whether this same pattern decreases the incidence of postoperative pulmonary complications (PPC) and hospital stay.

PATIENTS AND METHODS: This was a prospective, randomized clinical study. Between January 2012 and June 2013, 50 patients who had undergone lung resection with posterolateral thoracotomy were assigned to one of two groups by a random number generator according to whether or not they would receive prophylactic BiPAP pre- and postoperatively.

RESULTS: The results of the gasometric and spirometric values were similar in both groups. There were no statistically significant differences (p > 0.05). There was not a decrease in the incidence of PPC in the group that received prophylactic BiPAP. Likewise, postoperative stay was similar in both groups. The BiPAP group was 6.60 ± 4 days and the non BiPAP group was 6.84 ± 3.94 days (p = 0.63).

CONCLUSIONS: One drawback of this work was the limited number of hours that BiPAP was employed, and when compared to other studies, the application of low-pressure support. We did not find any significant differences between using prophylactic BiPAP or not, suggesting that such treatment should not be performed indiscriminately. More investigations are needed with a larger number of patients in order to better evaluate the possible benefits of using prophylactic BiPAP in thoracic surgery.

L'articolo Prophylactic use of non-invasive mechanical ventilation in lung resection sembra essere il primo su European Review.

http://ift.tt/2DewuPz

Correlations between CXCL13, IL-24 genes and wrist arthritis

OBJECTIVE: To investigate the relationship between B lymphocyte chemokine 1 (CXCL13) and interleukin-24 (IL-24) gene and wrist arthritis.

PATIENTS AND METHODS: A total of 122 cases of patients with wrist arthritis treated in our hospital from May 2013 to April 2016 were randomly selected as wrist arthritis group, while 120 normal subjects were selected as normal control group. Venous blood was collected from all patients in normal control group and wrist arthritis group, respectively. Rheumatoid factor (RF), human C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR) in venous blood were measured. The visual analogue scale (VAS) score was used to statistically analyze the pain of subjects in normal control and wrist arthritis groups; the wrist flexion and extension activities of subjects in normal control group and wrist arthritis group were measured. The expressions of CXCL13 and IL-24 mRNA in synovial tissue of normal control group and wrist arthritis group were detected by reverse transcription-polymerase chain reaction (RT-PCR). Western blotting was used to detect the expressions of CXCL13 and IL-24 in normal control group and wrist arthritis group.

RESULTS: The levels of CRP, RF, and ESR in the normal control group were within the normal range, but the levels of CRP, RF, and ESR in the wrist arthritis group were significantly higher than those in the normal control group. VAS scores and joint flexion extension activities in the normal control group were at normal levels. The VAS score of wrist arthritis group was significantly higher than that of the normal control group, and the joint flexion extension activities were significantly lower than that in the normal control group. The results of RT-PCR showed that the expression of CXCL13 mRNA in synovial tissue of wrist arthritis was significantly higher than that in the normal control group, while the expression of IL-24 mRNA in synovial tissue of wrist arthritis was significantly lower than that in normal control tissues. Western blotting showed that the expression of CXCL13 in synovial tissue of wrist arthritis was significantly higher than that in the normal control group, while the expression of IL-24 in synovial tissue of wrist arthritis was significantly lower than that in normal control groups. Analysis of variance showed that the expressions of CXCL13 and IL-24 in the normal control group and wrist arthritis group had statistically significant differences (p<0.01).

CONCLUSIONS: The abnormal expressions of CXCL13 and IL-24 are closely related to the occurrence and development of wrist arthritis. This study shows that CXCL13 and IL-24 have important research values in wrist arthritis. CXCL13 and IL-24 expressions can be used as new indicators of the diagnosis and treatment of wrist arthritis.

L'articolo Correlations between CXCL13, IL-24 genes and wrist arthritis sembra essere il primo su European Review.

http://ift.tt/2DlZqIs