1.     I have reviewed this Annual Report on Form 10-K for the year ended December 31, 2018 of Ra Pharmaceuticals, Inc.;

authority were to require us to conduct clinical trials beyond those that we currently anticipate will be required for the completion of clinical development, or if we experience significant delays in execution of or enrollment in any of our pre-clinical studies or clinical trials, we could be required to expend significant additional financial resources and time on the completion of pre-clinical and clinical development. We expect our research and development expenses to increase for the foreseeable future as we continue the development of product candidates.

ansonsaw/E+/GETTY IMAGES Continuous flow manufacturing for small-molecule drug synthesis received a vote of confidence in April 2016 when FDA approved a change from batch to continuous processing for Janssen Products’ Prezista (darunavir), a drug for the treatment of HIV-1 infection. This decision is the first such approval for a change from batch to continuous manufacturing, and the second for a continuous process; Vertex received approval in July 2015 for the continuous production of its cystic fibrosis drug Orkambi (lumacaftor/ivacaftor).

Cell proliferation was determined by cell viability assay and EdU incorporation assay. Approximately 4 × 103 cells were seeded in triplicate in 96-well plates. Cell viability was assessed using the CellTiter 96 Aqueous One Solution Proliferation Assay Kit (Promega). EdU labeling was performed using the EdU Assay Kit (Ribobio, Guangzhou, China) as recommended by the manufacturer. Cell migration was determined by wound-healing assay as previously described7 by measuring the decreased width of the scratch.

BioDelivery Sciences International Inc. (NasdaqCM: BDSI ) BioDelivery Sciences International, Inc. is a specialty pharmaceutical company that is focused on developing innovative products to address growing market opportunities, including conditions such as pain. The company utilizes its owned and licensed patented drug delivery technologies to develop, partner, and commercialize new products using proven therapeutics. BDSI’s pain franchise currently consists of two products in development utilizing the Company’s patented BEMA buccal soluble film technology: ONSOLIS, a potential treatment for “breakthrough” pain in opioid tolerant patients with cancer, and BEMA Buprenorphine, a second analgesic with at least one potential target indication for the treatment of moderate to severe pain. The company is working with its BEMA technology and its patented Bioral® cochleate technology on products targeted at conditions common to oncology and surgical patients such as pain and infections.

ArQule Inc. (NasdaqGM:ARQL) is a biotechnology company engaged in the research and development of next-generation, small-molecule cancer therapeutics. The Company’s targeted, broad-spectrum products and research programs are focused on key biological processes that are central to human cancers. ArQule’s lead product candidate, in Phase 2 and Phase 3 clinical development together with development and commercialization partner, Daiichi Sankyo, Co. Ltd., is tivantinib, an oral, selective inhibitor of the c-MET receptor tyrosine kinase. The Company’s pipeline consists of ARQ 621, designed to inhibit the Eg5 kinesin motor protein, and ARQ 736, designed to inhibit the RAF kinases. ArQule’s current discovery efforts, which are based on the ArQule Kinase Inhibitor Platform (AKIP™), are focused on the identification of novel kinase inhibitors that are potent, selective and do not compete with ATP (adenosine triphosphate) for binding to the kinase.

Evolva SA (SwissSIX:EVE) is an international, innovative synthetic biology company with a world-class research platform. Evolva strives to improve people’s lives by applying its technology and other resources to the discovery and development of new products and processes that benefit the health, well-being and financial economy of patients, consumers and partner companies around the world. Evolva uses biosynthetic and evolutionary technologies to artificially create and optimise small molecule compounds and their production routes. Our approach differs from that of the mainstream in the pharmaceutical and chemical industries. We have discovery partnerships ongoing both in pharma and industrial areas. In addition we have a pipeline of promising compounds aimed at infectious and cardio-renal indications

Well the EDQM has now added their word to the debate of contaminantshttps://www.edqm.eu/en/news/update-edqm-review-cep-applications-sartan-substances-4-february-2019

I could see CR being superior to IF in regards to total amount of mTOR activation but this is just speculation and i could see the opposite too.

To determine if the effect of cholesterol modulation on HCN1 channels could be generalized to other human HCN channel isoforms, we further examined the effects on the other cardiac isoforms, HCN2 and HCN4 (Figs 2 and 3). Intriguingly, we observed differential effects of cholesterol modulation on these isoforms. Similar to HCN1, both HCN2 and HCN4 channels showed a decrease in current density upon cholesterol depletion by either MβCD (Fig. 2A,B; Fig. 3A,B) or mevastatin (Supp. Fig. 1B,C). Moreover, current densities in cells expressing these isoforms enriched with cholesterol remained similar to control (Fig. 2A,B; Fig. 3A,B). HCN2 channels showed no differences in steady-state activation properties with changes in cholesterol content, however, steady-state properties of HCN4 channels were shifted approximately +10 mV by either cholesterol depletion or enrichment (Fig. 3C; Table 1). Tail currents were too small in HCN4 expressing cells treated with mevastatin to reliably enable us to determine steady-state activation properties. Intriguingly, the effects of cholesterol modulation on human HCN2 and HCN4 kinetics differed from our observations in human HCN1 channels. The activation kinetics of HCN2 and HCN4 channels were unaffected by cholesterol modulation (Figs 2D and 3D,E). However, unlike HCN1 channels, the deactivation kinetics of HCN2 and HCN4 channels were slowed by cholesterol enrichment (Figs 2E and 3F). These data suggest the effect of cholesterol on HCN channels is isoform specific.

We further examined voltage hysteresis in HCN channels using ramp protocols of various rates between 600 mV/s and 37.5 mV/s. We quantified the degree of hysteresis by the difference in area under the curves in the forward and reverse direction. We observed that for HCN1 channels, hysteresis increased with more rapid ramp speeds, and that manipulation of cholesterol levels in the membrane had no effect on the amount of hysteresis (Fig. 6A). On the other hand, HCN2 channels showed increased hysteresis as ramp speeds slowed. Cholesterol enrichment had no statistically significant consequences on the amount of hysteresis we observed for HCN2 channels, however, cholesterol depletion by MβCD and particularly mevastatin resulted in a drastic increase in the amount of hysteresis HCN2 channels underwent (Fig. 6B). This was particularly evident at 600 mV/s ramps in which in control membranes the current from 0 mV to −150 mV overlapped with the trace from −150 mV to 0 mV, while a large separation could be observed in all traces of cholesterol depleted cells expressing HCN2. Lastly, hysteresis was observed for HCN4 channels, however, the degree was unaffected by changes in ramp speeds. Moreover, hysteresis in HCN4 channels were unaffected by modulation of cholesterol levels (Fig. 6C). Taken together, these data indicate that cholesterol modifies hysteresis properties in HCN channels in an isoform specific manner.

Sorescu, D. & Griendling, K. K. Reactive oxygen species, mitochondria, and NAD(P)H oxidases in the development and progression of heart failure. Congest Heart Fail 8, 132–140 (2002).