Inhibition of the mammalian target of rapamycin (mTOR) in advanced pancreatic cancer: results of two phase II studies

<p>Abstract</p> <p>Background</p> <p>The phosphoinositide 3-kinase (PI3K)/Akt pathway is constitutively activated in pancreatic cancer and the mammalian target of rapamycin (mTOR) kinase is an important mediator for its signaling. Our recent <it>in vitro </it>studies suggest that prolonged exposure of pancreatic cancer cells to mTOR inhibitors can promote insulin receptor substrate-PI3K interactions and paradoxically increase Akt phosphorylation and cyclin D1 expression in pancreatic cancer cells (negative feedback loop). The addition of erlotinib to rapamycin can down-regulate rapamycin-stimulated Akt and results in synergistic antitumor activity with erlotinib in preclinical tumor models.</p> <p>Methods</p> <p>Two studies prospectively enrolled adult patients with advanced pancreatic cancer, Eastern Cooperative Oncology Group performance status 0-1, adequate hematologic, hepatic and renal parameters and measurable disease. In Study A, temsirolimus was administered intravenously at 25 mg weekly. In Study B, everolimus was administered orally at 30 mg weekly and erlotinib was administered at 150 mg daily. The primary endpoint in both studies was overall survival at 6 months. Secondary endpoints included time to progression, progression-free survival, overall survival, response rate, safety and toxicity. Pretreatment tumor biopsies were analyzed by immunofluorescence and laser scanning cytometry for the expression of pmTOR/mTOR, pAkt/Akt, pErk/Erk, pS6, p4EBP-1 and PTEN.</p> <p>Results</p> <p>Five patients enrolled in Study A; Two patients died within a month (rapid disease progression and hemorrhagic stroke, respectively). One patient developed dehydration and another developed asthenia. Sixteen patients enrolled in Study B.: 12 males, all ECOG PS = 1. Median cycles = 1 (range 1-2). Grade 4 toxicity: hyponatremia (n = 1), Grade 3: diarrhea (n = 1), cholangitis (n = 3), hyperglycemia (n = 1), fatigue (n = 1). Grade 2: pneumonia (n = 2), dehydration (n = 2), nausea (n = 2), neutropenia (n = 1), mucositis (n = 2) & rash (n = 2). Four patients were hospitalized. Progressive disease occurred in 15 and 1 was non-evaluable. Pretreatment biopsies revealed a higher pAkt/Akt ratio in tumor specimens that in nonmalignant pancreatic tissue. No such trends were noted for the other biomarkers.</p> <p>Conclusions</p> <p>Neither study with mTOR inhibitors demonstrated objective responses or disease stability. The negative feedback loop resulting from mTOR inhibition may account for the disease progression and toxicity noted in these studies. Future strategies should aim for a broader targeting of the PI3K pathway in pancreatic cancer.</p> <p>Trial Registration</p> <p><b>Trial registration: Study A</b>: NCT 0075647. <b>Study B</b>: NCT00640978</p

Quality Assessment in Crowdsourced Indigenous Language Transcription

The digital Bleek and Lloyd Collection is a rare collection that contains artwork, notebooks and dictionaries of the indigenous people of Southern Africa. The notebooks, in particular, contain stories that encode the language, culture and beliefs of these people, handwritten in now-extinct languages with a specialised notation system. Previous attempts have been made to convert the approximately 20000 pages of text to a machine-readable form using machine learning algorithms but, due to the complexity of the text, the recognition accuracy was low. In this paper, a crowdsourcing method is proposed to transcribe the manuscripts, where non-expert volunteers transcribe pages of the notebooks using an online tool. Experiments were conducted to determine the quality and consistency of transcriptions. The results show that volunteeers are able to produce reliable transcriptions of high quality. The inter-transcriber agreement is 80% for |Xam text and 95% for English text. When the |Xam text transcriptions produced by the volunteers are compared with a gold standard, the volunteers achieve an average accuracy of 64.75%, which exceeded that in previous work. Finally, the degree of transcription agreement correlates with the degree of transcription accuracy. This suggests that the quality of unseen data can be assessed based on the degree of agreement among transcribers

A recombinant Fasciola gigantica 14-3-3 epsilon protein (rFg14-3-3e) modulates various functions of goat peripheral blood mononuclear cells

Background The molecular structure of Fasciola gigantica 14-3-3 protein has been characterized. However, the involvement of this protein in parasite pathogenesis remains elusive and its effect on the functions of innate immune cells is unknown. We report on the cloning and expression of a recombinant F. gigantica 14-3-3 epsilon protein (rFg14-3-3e), and testing its effects on specific functions of goat peripheral blood mononuclear cells (PBMCs). Methods rFg14-3-3e protein was expressed in Pichia pastoris. Western blot and immunofluorescence assay (IFA) were used to examine the reactivity of rFg14-3-3e protein to anti-F. gigantica and anti-rFg14-3-3e antibodies, respectively. Various assays were used to investigate the stimulatory effects of the purified rFg14-3-3e protein on specific functions of goat PBMCs, including cytokine secretion, proliferation, migration, nitric oxide (NO) production, phagocytosis, and apoptotic capabilities. Potential protein interactors of rFg14-3-3e were identified by querying the databases Intact, String, BioPlex and BioGrid. A Total Energy analysis of each of the identified interaction was performed. Gene Ontology (GO) enrichment analysis was conducted using Funcassociate 3.0. Results Sequence analysis revealed that rFg14-3-3e protein had 100% identity to 14-3-3 protein from Fasciola hepatica. Western blot analysis showed that rFg14-3-3e protein is recognized by sera from goats experimentally infected with F. gigantica and immunofluorescence staining using rat anti-rFg14-3-3e antibodies demonstrated the specific binding of rFg14-3-3e protein to the surface of goat PBMCs. rFg14-3-3e protein stimulated goat PBMCs to produce interleukin-10 (IL-10) and transforming growth factor beta (TGF-β), corresponding with low levels of IL-4 and interferon gamma (IFN-γ). Also, this recombinant protein promoted the release of NO and cell apoptosis, and inhibited the proliferation and migration of goat PBMCs and suppressed monocyte phagocytosis. Homology modelling revealed 65% identity between rFg14-3-3e and human 14-3-3 protein YWHAE. GO enrichment analysis of the interacting proteins identified terms related to apoptosis, protein binding, locomotion, hippo signalling and leukocyte and lymphocyte differentiation, supporting the experimental findings. Conclusions Our data suggest that rFg14-3-3e protein can influence various cellular and immunological functions of goat PBMCs in vitro and may be involved in mediating F. gigantica pathogenesis. Because of its involvement in F. gigantica recognition by innate immune cells, rFg14-3-3e protein may have applications for development of diagnostics and therapeutic interventions

Oral Abstracts 7: RA ClinicalO37. Long-Term Outcomes of Early RA Patients Initiated with Adalimumab Plus Methotrexate Compared with Methotrexate Alone Following a Targeted Treatment Approach

Background: This analysis assessed, on a group level, whether there is a long-term advantage for early RA patients treated with adalimumab (ADA) + MTX vs those initially treated with placebo (PBO) + MTX who either responded to therapy or added ADA following inadequate response (IR). Methods: OPTIMA was a 78- week, randomized, controlled trial of ADA + MTX vs PBO + MTX in MTX-naïve early (<1 year) RA patients. Therapy was adjusted at week 26: ADA + MTX-responders (R) who achieved DAS28 (CRP) <3.2 at weeks 22 and 26 (Period 1, P1) were re-randomized to withdraw or continue ADA and PBO + MTX-R continued randomized therapy for 52 weeks (P2); IR-patients received open-label (OL) ADA + MTX during P2. This post hoc analysis evaluated the proportion of patients at week 78 with DAS28 (CRP) <3.2, HAQ-DI <0.5, and/or ΔmTSS ≤0.5 by initial treatment. To account for patients who withdrew ADA during P2, an equivalent proportion of R was imputed from ADA + MTX-R patients. Results: At week 26, significantly more patients had low disease activity, normal function, and/or no radiographic progression with ADA + MTX vs PBO + MTX (Table 1). Differences in clinical and functional outcomes disappeared following additional treatment, when PBO + MTX-IR (n = 348/460) switched to OL ADA + MTX. Addition of OL ADA slowed radiographic progression, but more patients who received ADA + MTX from baseline had no radiographic progression at week 78 than patients who received initial PBO + MTX. Conclusions: Early RA patients treated with PBO + MTX achieved comparable long-term clinical and functional outcomes on a group level as those who began ADA + MTX, but only when therapy was optimized by the addition of ADA in PBO + MTX-IR. Still, ADA + MTX therapy conferred a radiographic benefit although the difference did not appear to translate to an additional functional benefit. Disclosures: P.E., AbbVie, Merck, Pfizer, UCB, Roche, BMS—Provided Expert Advice, Undertaken Trials, AbbVie—AbbVie sponsored the study, contributed to its design, and participated in the collection, analysis, and interpretation of the data, and in the writing, reviewing, and approval of the final version. R.F., AbbVie, Pfizer, Merck, Roche, UCB, Celgene, Amgen, AstraZeneca, BMS, Janssen, Lilly, Novartis—Research Grants, Consultation Fees. S.F., AbbVie—Employee, Stocks. A.K., AbbVie, Amgen, AstraZeneca, BMS, Celgene, Centocor-Janssen, Pfizer, Roche, UCB—Research Grants, Consultation Fees. H.K., AbbVie—Employee, Stocks. S.R., AbbVie—Employee, Stocks. J.S., AbbVie, Amgen, AstraZeneca, BMS, Celgene, Centocor-Janssen, GlaxoSmithKline, Lilly, Pfizer (Wyeth), MSD (Schering-Plough), Novo-Nordisk, Roche, Sandoz, UCB—Research Grants, Consultation Fees. R.V., AbbVie, BMS, GlaxoSmithKline, Human Genome Sciences, Merck, Pfizer, Roche, UCB Pharma—Consultation Fees, Research Support. Table 1.Week 78 clinical, functional, and radiographic outcomes in patients who received continued ADA + MTX vs those who continued PBO + MTX or added open-label ADA following an inadequate response ADA + MTX, n/N (%)a PBO + MTX, n/N (%)b Outcome Week 26 Week 52 Week 78 Week 26 Week 52 Week 78 DAS28 (CRP) <3.2 246/466 (53) 304/465 (65) 303/465 (65) 139/460 (30)*** 284/460 (62) 300/460 (65) HAQ-DI <0.5 211/466 (45) 220/466 (47) 224/466 (48) 150/460 (33)*** 203/460 (44) 208/460 (45) ΔmTSS ≤0.5 402/462 (87) 379/445 (86) 382/443 (86) 330/459 (72)*** 318/440 (72)*** 318/440 (72)*** DAS28 (CRP) <3.2 + ΔmTSS ≤0.5 216/462 (47) 260/443 (59) 266/443 (60) 112/459 (24)*** 196/440 (45) 211/440 (48)*** DAS28 (CRP) <3.2 + HAQ-DI <0.5 + ΔmTSS ≤0.5 146/462 (32) 168/443 (38) 174/443 (39) 82/459 (18)*** 120/440 (27)*** 135/440 (31)** aIncludes patients from the ADA Continuation (n = 105) and OL ADA Carry On (n = 259) arms, as well as the proportional equivalent number of responders from the ADA Withdrawal arm (n = 102). bIncludes patients from the MTX Continuation (n = 112) and Rescue ADA (n = 348) arms. Last observation carried forward: DAS28 (CRP) and HAQ-DI; Multiple imputations: ΔmTSS. ***P < 0.001 and **iP < 0.01, respectively, for differences between initial treatments from chi-squar

Distribution and phylogenetics of hepatitis E virus genotype 4 in humans and animals

BackgroundWorldwide, hepatitis E virus (HEV) infection is considered a significant public health concern. In particular, HEV genotype 4 (HEV-4) has spread to more areas and host species. In this study, we describe the global distribution of HEV-4 and characterize HEV-4 subtypes by host, country and year of isolation.MethodsWe retrospectively collected HEV-4 sequences available before December 31, 2019, in GenBank. HEV-4 and its subtypes were determined using phylogenetic comparison with HEV reference sequences. Information on the isolation of the sequences was extracted from the GenBank or original publications. Temporal, spatial and host characteristics of the sequences were summarized and nucleotide similarity was calculated based on five amplified fragments within HEV genome, stratified by host, country and year.ResultsA total of 2295 HEV-4 complete and partial nucleotide sequences were studied. The majority (92.7%) was isolated in China’s mainland, Japan, Hong Kong and France. A total of 20 animal hosts were documented, though swine remained predominant (71.7%). Globally, prevalent HEV-4 subtypes changed remarkably over the last 18 years. Subtypes 4a, 4b, 4d and 4h were most commonly isolated (80.3%). Subtypes 4c, 4e, 4f, 4g and 4i remained limited in temporal distribution. High nucleotide similarities were observed between the sequences amplified in HEV ORF2, in the same and neighbouring countries, and in similar animal hosts.ConclusionChina and Japan are endemic for HEV-4, and have all the subtypes. In Europe, France has a high prevalence of HEV-4. Increases in affected areas and animal hosts imply consistent cross-border and cross-species transmission.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/173100/1/zph12934_am.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/173100/2/zph12934.pd

Associations between health indicators and sleep duration of American adults:NHANES 2011-16

Background: This study aimed to investigate associations between health indicators and sleep duration in the general population. Methods: This cross-sectional data from the National Health and Nutrition Examination Survey. Self-reported sleep duration was classified into short sleep (&lt;7 h/day), regular sleep (7-8 h/day) and long sleep duration (&gt;8 h/day). Health indicators included lifestyle indicators (smoking, alcohol use and physical inactivity), general health indicators (waist circumference and self-reported health condition) and chronic conditions [overweight/obesity, hypertension, diabetes, high cholesterol, chronic low back pain (CLBP) and oral health problems]. A series of multinomial logistic regression analysis were performed, controlling for confounders (age, sex, marital status, ethnic background, education level and poverty-to-income ratio). Results: Data of 12 835 participants were analyzed. The mean (SD) age of participants was 50.0 (±17.4) years, and 50.6% were women. After adjusting for all health indicators, current smoking (OR: 1.37; 95% CI: 1.17-1.61), a poor (OR: 1.52; 95% CI: 1.23-1.88) health condition, CLBP (OR: 1.40; 95% CI: 1.16-1.69) and oral health problems (OR: 1.28; 95% CI: 1.10-1.49) were associated with short sleep duration. No independent association with long sleep duration was observed in this study. Conclusions: The results confirm that lifestyle indicators (current smoking and physical inactivity), general health indicators (self-reported health condition) and presence of some chronic conditions (CLBP and oral health problems) are associated with short sleep duration. The results did not confirm that any health indicator was associated with long sleep duration.</p