The protein ubiquitin is an important post-translational modifier that regulates a wide variety of biological processes. In cells, ubiquitin is apportioned among distinct pools, which include a variety of free and conjugated species. Although maintenance of a dynamic and complex equilibrium among ubiquitin pools is crucial for cell survival, the tools necessary to quantify each cellular ubiquitin pool have been limited. We have developed a quantitative mass spectrometry approach to measure cellular concentrations of ubiquitin species using isotope-labeled protein standards and applied it to characterize ubiquitin pools in cells and tissues. Our method is convenient, adaptable and should be a valuable tool to facilitate our understanding of this important signaling molecule.

Background: Polyglutamine aggregates can be internalized by mammalian cells and gain access to the cytoplasmic compartment, but the properties of the aggregates and cell surface that mediate these processes are unknown. Results: Introduction of net negative charge and disruption of fibrillar structure greatly reduced the capacity of polyglutamine aggregates to bind and be internalized by mammalian cells. Conclusion: Aggregate uptake is influenced by the structure and net charge of aggregates and is mediated by two classes of binding sites on the cell surface. Significance: Elucidating how protein aggregates are internalized by cells is important for understanding the pathogenesis of many neurodegenerative disorders.

A hypothetical scheme is presented to delineate the plausible cause for the slump of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) ‐ Cas9: “CRISPR‐associated protein 9”) labyrinthine gene editing of ARGR would gleam a catastrophic augmentation leading to an evolution of virulent bacterial pathogen(s) dithering sensitivity to antibiotics, impairing the effective treatment of antibiotic resistance bacterial pathogens induced sepsis, exacerbating disseminated intravascular coagulation (DIC) unto antibiotic resistance pandemic (ARP). Given the fact that AR is developmental malady enthralling intensive critical care inclusive of fluid resuscitation in a clinical emergency (patient cohort of sepsis treated with EGDT trial NCT01663701 resulting in‐hospital mortality.) A comparative model as intricate as CRISPR‐Cas‐9, while as sacrosanct as Chakravyūha: Chakra (spinning wheel), a strategy: (“vyuh”), with the lotus flowerets (Padmavyuh) made of seven circular paths characteristic of constant movement to that of a spiral down pattern of clustered regularly interspaced paths to the center core with strongest forces to mount resistance compare to that of the first layer with entrance ( Exhibit B1: https://www.youtube.com/watch?v=oy3Qzt74V6Q; Exhibit B2: https://youtu.be/hHc705uTOWc) was analyzed to derive an imperative of gene editing and its amelioration of antibiotic resistance. As suggested by the presenters, the formulation to break Chakravyūha is to increase the deployment of force in the following format per each path: force deployment per path would be 1/7 = 0.142857 where the denominator = total paths; whereas numerator is the specific number of path in the Chakravyūha. Therefore for path one force required to break and/or to overcome the resistance is = 1/7 = 0.142857142857; for path two = 0.142857 × 2 = 0.28571142857; for path three 0.142857 × 3= 0.428571142857; for path four 0.142857 × 4= 0.57142857142857; for path five = 0.142857 × 5 = 0.7142857142857; for path 0.142857 × 6 = 0.857142857142857; for path seven = 0.142857 × 7 = 0.857142857142857; and path seven @ core = 0.142857 × 7 = 0.999999142857 considered rounding off to 1. The unique six digits, 142857 is repeated among all paths similar to the basis of gene editing compare the Exhibit A: time 0.55 – 3.50 min; to that of the Exhibit B1: time:6.11 – 7.25 min & 2: Exhibit B2 time 5.51 – 7.25 for mitigating the AR by targeting and editing of antibiotic resistance genes (ARG) in ARGR would lead an increased inactivation of preexisting ARG conferring sensitivity to antibiotics benefiting the antibiotic therapy but not clinically evolving AR which is predominantly regulated by infection specific antibiotic regimen of a patient’s life span tacit of multifactorial inclusive of exposure to multitude AR bacteria (ARB) at different stages of life span, and patient specific clinical history. Treatment options where in which inherent off target errors of gene editing by CRISPR‐Cas9’s and inability to match the rate of commission versus correction [t...