9 However, in some instances, the etiological root of a disease may begin in the ENS and spread to other areas of the body. Many of these disorders are not life threatening but they do cause extreme patient discomfort and reduced quality of life. 8Ībnormalities of the ENS cause disruptions in GI function that manifest in functional disorders such as gastroparesis, irritable bowel syndrome, Allgrove syndrome, hypertrophic pyloric stenosis, Hirschprung disease, or intestinal neuronal dysplasia. Although the ENS is connected to the CNS, the ENS is composed of integrative neuronal circuits, which can control most gut functions independent of CNS connections. The myenteric plexus is located between the circular and longitudinal muscle, while the submucosal plexus is located just below the mucosa in the lumen of the gut. These neurons are organized into ganglia located in either the myenteric plexus (MP) or submucosal plexus (SMP). The ENS consists of an estimated 100 million neurons, which are embedded in the gut wall, spanning from the esophagus to the anus. The ENS coordinates control of enterocyte secretion, nutrient and fluid absorption, blood flow, and motor function in the gastrointestinal (GI) system. The ENS is a division of the autonomic nervous system. Although there have been reports that AAV can transduce cells of the gut following systemic or peroral delivery, 5, 6, 7 to date, there has been no focused investigation on the ability of AAV to directly transduce cells of the enteric nervous system (ENS) following targeted injections. 2, 4 AAV vectors can efficiently transduce many different biological tissues including muscle, liver, brain, lung, and retina (reviewed in ref. Further, AAV vectors can achieve high levels of transduction within weeks that are maintained over the lifetime of an organism. 1, 2, 3 AAV is favored over other viral vectors due to its lack of pathogenicity and the ability to establish expression within both dividing and mature nondividing cells of multiple lineages. Recombinant adeno-associated virus (AAV) is one of the most widely used gene therapy systems in research and clinical trials. This is the first report on targeted AAV transduction of neurons and glia in the ENS. AAV9 primarily transduced neurons, while AAV6 transduced enteric glia and neurons. A single injection of AAV into the colon covered an area of ~47 mm 2. Transduction efficiency scaled with titer and time, was translated to the murine ENS, and produced no vector-related immune response. AAV6 and AAV9 showed the highest levels of transduction within the ENS. Transduction, as determined by GFP-positive cells, occurred in neurons and enteric glia within the myenteric and submucosal plexuses of the ENS. AAV serotypes tested included AAV 1, 2, 5, 6, 8, or 9 and the AAV2 and AAV8 capsid mutants, AAV2-Y444F, AAV2-tripleY-F, AAV2-tripleY-F+T-V, AAV8-Y733F, and AAV8-doubeY-F+T-V. Rats received direct injections of various AAV serotypes expressing green fluorescent protein (GFP) into the descending colon. Here, we examined the efficiency, tropism, spread, and immunogenicity of AAV transduction in the ENS. AAV can transduce a wide range of biological tissues, however to date, there has been no investigation on targeted AAV transduction of the enteric nervous system (ENS). I’m still working on my weight loss but I’m down 20 pounds so far.Recombinant adeno-associated virus (AAV) vectors are one of the most widely used gene transfer systems in research and clinical trials. That’s going to do more for you than a pink drink. Eat food, mostly plants, some meat and eggs and fish if you want, and drink lots of water. You don’t have to starve yourself or drink chemicals. A nutritionist will get you set up with a good calorie counting regimen and a bunch of recipes and good advice and help you along the way. I think many other companies do the same. My insurance covers nutrition counseling 100 percent because it prevents future health issues and saves them money. You’re better off talking to a nutritionist. If I’d continued it, it could have been worse. It felt like I was running a marathon and I felt lightheaded. I’m lucky nothing happened because I have a genetic heart condition that wasn’t officially diagnosed until a year later. You may have a health condition you don’t know about and it can be exacerbated by these things. I would not recommend trying this or anything similar to it.
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