Longevity Questions

Perpetual Prompt ♾️, [08.05.2025 23:57]
901.What is the minimum duration of normothermic machine perfusion (NMP) of the liver with the MSC-bioreactor, allowing to reduce the ALT/AST activity in the perfusate to ≤50% of the initial level after 12 hours of warm ischemia?

902.What is the target number of mesenchymal stromal cells per 1 ml of perfusate that provides ≥70% suppression of HMGB1 release in the first 180 minutes of NMP?

903.At what flow rate (ml min⁻¹ 100 g⁻¹ tissue) is the mitochondrial respiratory control ratio (RCR) ≥7 achieved in porcine liver after 4 hours of NMP?

904.What ratio of glucose to ketone bodies in the perfusion medium maximizes bile production (≥0.6 ml g⁻¹ h⁻¹) in a rat liver model?

905.What range of liver core temperature (°C) during LMP is optimal to maintain tissue ATP levels ≥4 µmol g⁻¹?

906.What dose of MSC exosomes (µg protein ml⁻¹) inhibits caspase-3 in hepatocytes to ≤20% of control value within 2 h of LMP?

907.What pO₂ level (mmHg) in the perfusate is required to reduce mitochondrial ROS formation to <5pmol min⁻¹ mg⁻¹ protein?

908.What concentration of NO-donors (µM) should be maintained in the circuit to ensure vascular resistance ≤250 dyn s cm⁻⁵ without hypotension?

909.What portal vein pressure (mmHg) results in the lowest albumin leak (<0.2g L⁻¹) from liver tissue during LMP?

910.What is the optimal rate of L-arginine addition (mmol h⁻¹) to prevent an increase in endothelin-1 in the perfusate above 10pg ml⁻¹?

911.What is the minimum titer of the anti-inflammatory cytokine IL-10 (pg ml⁻¹) in the perfusate required to reduce ICAM-1 expression on sinusoidal endothelial cells by ≥60%?

912.What NAD⁺/NADH ratio must be maintained in tissue extracts to ensure that the rate of β-oxidation of fatty acids exceeds 1.5 nmol min⁻¹ mg⁻¹ protein?

913.What is the upper limit of lactate concentration (mmol L⁻¹) in the perfusate at which the pH can be maintained ≥7.35 ​​without bicarbonate correction?

914.What pool of microbial metabolites (SCFA) and in what quantities (µmol L⁻¹) must enter the hepatic portal tract to activate the AMPK-SIRT1 pathway in hepatocytes?

915.What combination of antifibrotic factors (e.g., HGF, MMP-9) and their concentrations (ng ml⁻¹) prevents hepatic stellate cell activation to ≤10%?

916.What duration of liver tissue exposure to 5µM resveratrol in the NMP circuit is required to lengthen hepatocyte telomeres by ≥200bp per cycle?

917.What threshold level of the mitophagy marker PINK1 (fold-change) indicates complete restoration of the mitochondrial pool after 6 h of NMP?

918. What amino acid composition (g L⁻¹ of each amino acid) minimizes urea formation (<0.05 mmol L⁻¹ h⁻¹) while maintaining albumin synthesis?

919. What concentration of β-hydroxybutyrate (mmol L⁻¹) should be maintained to ensure FOXO3a expression in hepatocyte nuclei >1.8-fold of control?

920. What target level of malondialdehyde (MDA, nmol mg⁻¹ protein) is considered a sufficient indicator of minimal lipid peroxide stress after reoxygenation?

921. What is the maximum storage time of liver at 4°C (hours) acceptable before switching to NMP without increasing mitochondrial Δψm-variance by >25%?

922.What is the limit of free Ca²⁺ (µM) in the perfusate at which calpain-dependent proteolysis of structural proteins is not activated?

923.What glutathione concentration (mmol L⁻¹) should be maintained to maintain a GSH/GSSG ratio ≥5:1 in the cytosol of hepatocytes?

924.What volume of extracorporeal plasma exchange (ml kg⁻¹) during NMP is optimal for removing accumulated inflammatory mediators without losing coagulation factors?

925.What threshold index of DNA synthesis (BrdU-positive cells, %) reflects the initiation of regenerative proliferation after 24 h of NMP +MSC?

926.What density of oxygen nanosensors (pieces cm⁻² of catheter surface) should be used for continuous pO₂ monitoring with an accuracy of ±2 mmHg?

Perpetual Prompt ♾️, [08.05.2025 23:57]
927.What level of TERT mRNA (fold-change) expression is required in hepatocytes to predict a prolongation of cellular replicative life of ≥20 divisions?

928.What combination of microRNA-modulating oligonucleotides (names and nM) increases the expression of the antioxidant gene SOD2 by at least 3-fold?

929.What reduction in cold ischemia time (h) leads to a statistically significant (p<0.05) decrease in the incidence of early graft dysfunction to <5%?

930. What is the maximum acceptable residual endotoxin load (EU ml⁻¹) in the perfusion circuit that does not increase liver TNF‑α expression above 15pg ml⁻¹?

Perpetual Prompt ♾️, [08.05.2025 23:59]
931. What is the minimum number of CD34⁺CD38⁻CD90⁺CD45RA⁻ cells (per kg body weight) required to repopulate adult human bone marrow to ≥95% donor chimerism without complete myeloablation?
932. Does prior deletion of CD150^high^ (senescent) clones via targeted apoptosis reduce the level of epigenetic aging index in human blood ≥5 years per course of therapy (yes/no)?
933. What is the maximum acceptable dose of antibody-radionuclide conditioning therapy (mCi/m²) that preserves ≥80% of native mesenchymal stromal cells-"niche"?
934. How many ex vivo division cycles can young HSCs undergo at 5% O₂ to maintain the reticular methylation index <1% of the initial level?
935. Is the frequency of clonal hematopoiesis of indeterminate potential (CHIP) reduced to <0.1% after CRISPR correction of DNMT3A/TET2 mutations in old HSCs?
936. Does a single intraosseous injection of 10⁸ extracellular vesicles from young HSCs reduce the levels of proinflammatory cytokines IL-6 and TNF-α in plasma of old mice by ≥50% over 30 days?
937. Can 14-day exposure to the combination of SCF+TPO+UM171 increase the frequency of long-lived human HSC-LT (CD49f⁺) by >10-fold without increasing karyotypic aberrations?
938. Does short-term (48h) partial reprogramming (OSK-mRNA) in old human HSCs induce a decrease in epigenetic age by Horvath sequencing of ≥10 years without loss of multipotency?
939. Is the rejuvenating effect of young HSC transplantation in mice ≥24 months, as measured by the GrimAge-Clock, maintained without retransplantation (yes/no)?

940. Does administration of anti-CXCR4 antibodies after young HSC transplantation reduce the time to neutrophil recovery to <10 days in patients ≥65 years old?

941. What ratio of donor to assessed resident HSCs is required to suppress >90% of senescent CD150^high^ clones in primates?
942. Can a p38MAPK inhibitor administered in vivo after transplantation increase 5-year survival of donor HSCs by >30%?
943. What is the minimum number of HLA-matched loci required to keep the GVHD rate <5% after infusion of young allogeneic HSCs in non-myeloablated patients?
944. Is inhibition of NADPH oxidase4 sufficient to reduce reactive oxygen species in aged HSCs to <2μM and restore their colony-forming activity?
945. Does transient hyperbaric oxygenation (2 atm, 90 min) before HSC collection reduce the level of EGR1 promoter hypomethylation to ≤0.5%?
946. Is functional thymus mass restored (>50% of control young) in aged mice after removal of aged HSCs and infusion of young ones for 12 weeks?
947. What threshold level of mitochondrial potential (Δψm, mV) in donor HSCs correlates with >80% long-term repopulation in vivo?
948. Does delivery of siRNA against p16^Ink4a^ to aged HSCs reduce their senescent fraction below 5% without inducing apoptosis?
949. Does complete removal of circulating SASP factors (IL-1β, IL-8) by plasmapheresis reduce the incidence of myeloid malignant transformations in patients aged >75 years?
950. Is 3-D collagen-I matrix with a stiffness of 0.4 kPa required to preserve the transcriptome of young HSCs ex vivo better than standard 2-D plastic (p<0.05)?