RNA interference (RNAi) is a powerful tool for target identification and can lead to novel therapies for pharmacologically intractable targets such as KRAS. RNAi therapy must combine potent siRNA payloads with reliable in vivo delivery for efficient target inhibition. We employed a functional “Sensor” assay to establish a library of potent siRNAs against RAS pathway genes and show they efficiently suppress their targets at low dose. This reduces off-target effects and enables combination gene knockdown. We administered Sensor siRNAs in vitro and in vivo and validated the delivery of KRAS siRNA alone and siRNA targeting the complete RAF effector node (A/B/C-RAF) as promising strategies to treat KRAS-mutant colorectal cancer. We further demonstrate that improved therapeutic efficacy is achieved by formulating siRNA payloads that combine both single-gene siRNA and node-targeted siRNAs (KRAS+PIK3C-A/B). The customizable nature of Sensor siRNA payloads offers a universal platform for combination target identification and development of RNAi therapeutics.

Most biological systems depend on preservation of equilibrium. Within the male reproductive system, oxidative harmony is crucial to fertility. Reactive oxygen species (ROS) make functional contributions at appropriate concentrations, but quickly become destructive if left unchecked. It is important to understand the key oxidative players, their mechanisms and the implications in male infertility. In this review, we discuss the role of redox biology in male reproduction, consequences of oxidative stress and their involvement in pathogenesis of male infertility and assisted reproductive outcomes. 1.1 | Oxidative stress and sources of ROS Given the premise of aerobic life, reactive oxygen species play a paradoxical role, both critical for regulation of cellular function and capable of exerting significant damage on cells and molecules (Morrell, 2008). Oxidative stress is defined as an imbalance between ROS and the antioxidants attempting to keep them in check (Sies, 2015). ROS exist in both endogenous and exogenous forms. Endogenous players include superoxide (O − 2), generated via mitochondrial redox reactions, as well as hydrogen peroxide (H 2 O 2) and the hydroxyl radical (OH −). Mitochondrial dysfunction results in the excessive accumulation of these products (Hayyan, Hashim, & AlNashef, 2016). Furthermore, immune cells employ redox reactions as anti-pathogen tools, generating ROS in the process (Sies, 1993). Nitric oxide is one in a collection of reactive nitrogen species (RNS), which also include peroxynitrite, nitroxyl and nitrosyl compounds (Doshi, Khullar, Sharma, & Agarwal, 2012). The product of the reaction between nitric oxide and superoxide, peroxynitrite, exerts oxidative stress on spermatozoa via direct molecular damage and interference with key signalling pathways (Rosselli, Keller, & Dubey, 1998). In this way, ROS hold the power to instigate exponential destruction (Henkel, Samanta, & Agarwal, 2018). Exogenous sources of ROS include tobacco smoke, heavy metals and ionizing radiation (Birben, Sahiner, Sackesen, Erzurum, & Kalayci, 2012). Oxygen free radicals in cigarette tar and tobacco smoke,

Narcolepsy is a chronic sleep disorder characterized by excessive daytime sleepiness, cataplexy, hypnagogic hallucinations, and sleep paralysis. Both sporadic and familial forms exist in humans. Recently, the major pathophysiology of human narcolepsy was indicated, based on discovery, through animal study, of narcolepsy genes involved in the pathology of hypocretin/orexin ligand and its receptor. Hypocretin ligand deficiency is found in most patients with narcolepsy with cataplexy. This deficiency likely is the result of postnatal cell death of hypocretin neurons, and involvement of autoimmune mechanisms is suggested. Hypocretin deficiency also is found in symptomatic narcolepsy and excessive daytime sleepiness with neurologic conditions, including immune-mediated neurologic disorders. These findings have significant clinical relevance and promote understanding of hypocretin cell death mechanisms. Already, discoveries in humans have led to a new diagnostic test for narcolepsy. Currently, hypocretin replacement therapy has emerged as a promising therapeutic option, and experiments using gene therapy and cell transplantation are in progress.

Although unstripped laser fibers provided superior power output at 1 minute, output at all other time points was similar between stripped and unstripped fibers. However, despite similar optical output, stripped laser fibers achieved greater stone fragmentation, possibly due to improved contact between stone and fiber tip.