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This was a mixed design in which each rat received saline and two of five possible doses of GLP-1 (0.1, 0.3, ColonBroom capsules 0.6, 1.0, and 10 μg in 2 μl of saline) over the course of the experiment. 5−6 plates per experiment and observed at 24−48 h intervals to document live, dead or censored (animals that exploded, bagged or could not be located) animals. At the same time, a similar number of age-matched, nontransgenic siblings were collected for each strain and assayed for lifespan as internal controls in the experiment. For lifespan assays of strains with the temperature sensitive glp-1 mutation, eggs were picked and maintained at 20 °C for 2−4 h, transferred to 25 °C to induce sterility and then returned to 20 °C on day 1 of adulthood (72 h later) for lifespan analysis. The plates were then left to sit at room temperature (RT) for 24 h prior to use.

For performing lifespan assays of transgenic strains with extrachromosomal arrays, eggs were picked onto fresh OP50 plates, incubated at the appropriate temperature and 48 h later L4 animals were screened under a Leica M165FC microscope with a fluorescence attachment (Leica Microsystems, Wetzlar, Germany) for animals carrying the red coinjection marker labeling pharyngeal muscles. To analyze PA14 sensitivity of C. elegans strains at various stages of life, temporal assays were conducted by picking eggs of wild-type and tcer-1 strains on OP50 plates, growing at 20 °C and then transferring them onto PA14 SK plates at L4, day 2, day 4, day 6 or day 9 of adulthood and monitoring for survival at 25 °C as mentioned above. The tcer-1 coding region was amplified with primers modified to introduce SalI and Acc65I restriction sites (forward 5′gctagGGTCGACatgagccacgaaaatc3′; reverse 5′taagcaGGTACCTCttgctttctgcgatcccgc3′). nhr-49 coding region was removed from the tissue-specific promoter plasmids using restriction enzymes, SalI and Acc65I, and replaced with PCR amplified and digested tcer-1 coding region by ligation into the respective plasmids in frame with the GFP. To generate tissue-specific TCER-1 expressing constructs the 4 kb tcer-1 coding region was inserted into plasmids created previously in the lab for another gene, nhr-49 (Ratnappan and Ghazi, unpublished).

The plasmids thus obtained were pAG11 (a 2.4 kb muscle promoter76, Pmyo-3::TCER-1::GFP), pAG12 (1.9 kb intestinal promoter77, Pgly-19:: TCER-1::GFP), pAG13 (400 bp hypodermal promoter78, Pcol-12:: TCER-1::GFP), and pAG14 (4.2 kb region neuronal promoter79, ColonBroom capsules Prgef-1:: TCER-1::GFP). Transgenic strains were generated by injecting the plasmids at a concentration of 25 ng per μl or 100 ng per μl along with 3.75 ng per μl or 15 ng per μl of Pmyo-2::mCherry co-injection marker, respectively. Three to six independent stable transgenic lines were generated for each of the genetic backgrounds in which the transgene was injected. Transgenic strains were maintained by picking fluorescent animals in each generation. All strains were grown and maintained on standard nematode growth medium (NGM) at 20 °C using E. coli strain OP50 as the food source. 20 µl of this broth culture was seeded onto slow killing (SK) plates (modified NGM plates containing 0.35% peptone instead of 0.25%) and incubated for 24 h at 37 °C. All lifespan experiments were conducted at 20 °C on E. coli OP50 plates unless otherwise noted. Each lifespan was tested at least twice and often in 3−5 biological replicates. The glucagon-like peptide-1 receptor (GLP-1R) is an important physiologic regulator of insulin secretion and a major therapeutic target for diabetes mellitus.

The major source of GLP-1 in the body is the intestinal L cell that secretes GLP-1 as a gut hormone. A body mass index (BMI is calculated by considering a person’s weight and height to measure body size) of more than 25 is considered overweight and more than 30 is considered obese. The NAD matter arose from a challenge brought by a branded manufacturer against a telehealth provider that marketed compounded semaglutide for weight loss. Across the reviews, tirzepatide, semaglutide, and liraglutide each led to significant weight loss compared to placebo after one to two years, with these effects likely to be sustained while treatment continues. GLP1/GLP-1R signaling continues upon entry into early endosomes. Today, the U.S. Food and Drug Administration approved the first generic referencing Victoza (liraglutide injection) 18 milligram/3 milliliter, a glucagon-like peptide-1 (GLP-1) receptor agonist indicated to improve glycemic control in adults and pediatric patients aged 10 years and older with type 2 diabetes as an adjunct to diet and exercise.