In some embodiments, calcium oscillations for a molecule with tolerable in vivo acute toxicity are greater than or equal to about 250%, greater than or equal to about 240%, greater than or equal to about 230%, greater than or equal to about 220%, greater than or equal to about 210%, greater than or equal to about 200% greater than or watch live sexcam equal to about 190%, greater than or equal to about 180%, greater than or equal to about 170%, greater than or equal to about 160%, greater than or equal to about 150%, greater than or equal to about 140%, greater than or equal to about 130%, greater than or equal to about 120%, greater than or equal to about 110%, greater than or equal to about 100%, greater than or equal to about 99%, greater than or equal to about 98%, greater than or equal to about 97%, greater than or equal to about 96%, greater than or equal to about 95%, greater than or equal to about 90%, greater than or equal to about 85%, greater than or equal to about 80%, greater than or equal to about 75%, or greater than or equal to about 70% of calcium oscillations in a vehicle control cell (e.g., water or saline).



In other embodiments, the calcium oscillations with tolerable in vivo acute toxicity are about 70% to about 250%, about 70% to about 200%, about 75% to about 200%, about 70% to about 180%, about 75% to about 150%, about 80% to about 200%, about 90% to about 200%, watch live Sexcam about 100% to about 200%, or about 80% to about 250% of the calcium oscillations in the vehicle control cells. In certain embodiments, the calcium oscillations with tolerable in vivo acute toxicity are greater than or equal to about 70% of the calcium oscillations in the vehicle control cells. In some embodiments, the calcium oscillations in a cell exposed to a molecule having a tolerable in vivo acute neurotoxicity exhibits less than about 25%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, or less than about 1% reduction compared to the calcium oscillations in a vehicle control cell.



In certain embodiments, the present disclosure includes a method of selecting a polynucleotide having tolerable in vivo acute neurotoxicity comprising (i) performing a calcium oscillation assay disclosed herein and (ii) calculating a sequence score disclosed herein, wherein the calcium oscillations of the polynucleotide are equal to or greater than 75% of the calcium oscillations in a vehicle cell and the sequence score of the polynucleotide is greater than or equal to 0.25. In some embodiments, the calcium oscillation assay and/or the sequence score method are sufficient to predict, identify, or determine in vivo acute neurotoxicity of a molecule and do not require additional in vivo tolerability studies. The present disclosure is also directed to a method of testing or determining in vivo neurotoxicity of a molecule (e.g., polynucleotide) comprising a nucleotide sequence. For Watch Live Sexcam example, a nucleotide sequence of ATGCATGCATGCATGC (SEQ ID NO: 3) has a sequence score of 0 ((4Cs-4Gs)/16). For example, molecules can be injected into a laboratory animal by ICV or IT. For example, long term toxicities can be determined by measuring the change in tubulin intensity in a cell by the molecule when the cell comes in contact with a molecule.



In certain embodiments, the methods of the disclosure further comprise measuring long term in vivo toxicities of a molecule. In other embodiments, the disclosure includes a method of identifying or selecting a molecule having tolerable in vivo neurotoxicity comprising (i) performing a calcium oscillation assay, (ii) calculating sequence score, (iii) performing an in vivo tolerability study, and (iv) administering the molecule to a mammal in need of treatment of a disease or condition. When the number of candidate molecules are small, an in vivo tolerability study can provide a direct indication of in vivo neurotoxicity. G nucleotides and analogs thereof Total nucleotide length ( number ) . Each category is scored on a scale of 0-4, with the worst possible total score of 20 and the best possible total score of 0. Animals are observed for changes in behavior, for example in the home cage, but they can be observed in other environments. G nucleotides Total nucleotide length . In certain embodiments, molecules comprising nucleotide sequences that have a sequence score below the set thresholds are considered to be molecules that have unacceptable neurotoxicity. Blondes have always been a very important part of porn since they are usually very naughty and eager to get messy with a good fucking.