Runx1 Messenger RNA Delivered by Polyplex Nanomicelles Alleviate Spinal Disc Hydration Loss in a Rat Disc Degeneration Model

Vertebral disc degenerative disease (DDD) affects millions of people worldwide and is a critical factor leading to low back and neck pain and consequent disability. Currently, no strategy has addressed curing DDD from fundamental aspects, because the pathological mechanism leading to DDD is still controversial. One possible mechanism points to the homeostatic status of extracellular matrix (ECM) anabolism, and catabolism in the disc may play a vital role in the disease’s progression. If the damaged disc receives an abundant amount of cartilage, anabolic factors may stimulate the residual cells in the damaged disc to secrete the ECM and mitigate the degeneration process. To examine this hypothesis, a cartilage anabolic factor, Runx1, was expressed by mRNA through a sophisticated polyamine-based PEG-polyplex nanomicelle delivery system in the damaged disc in a rat model. The mRNA medicine and polyamine carrier have favorable safety characteristics and biocompatibility for regenerative medicine. The endocytosis of mRNA-loaded polyplex nanomicelles in vitro, mRNA delivery efficacy, hydration content, disc shrinkage, and ECM in the disc in vivo were also examined. The data revealed that the mRNA-loaded polyplex nanomicelle was promptly engulfed by cellular late endosome, then spread into the cytosol homogeneously at a rate of less than 20 min post-administration of the mRNA medicine. The mRNA expression persisted for at least 6-days post-injection in vivo. Furthermore, the Runx1 mRNA delivered by polyplex nanomicelles increased hydration content by ≈43% in the punctured disc at 4-weeks post-injection (wpi) compared with naked Runx1 mRNA administration. Meanwhile, the disc space and ECM production were also significantly ameliorated in the polyplex nanomicelle group. This study demonstrated that anabolic factor administration by polyplex nanomicelle-protected mRNA medicine, such as Runx1, plays a key role in alleviating the progress of DDD, which is an imbalance scenario of disc metabolism. This platform could be further developed as a promising strategy applied to regenerative medicine.

Numerical investigation on the impact of multiple injection strategies and spray included angles on combustion and emissions in a marine diesel engine

The computational fluid dynamical software AVL-FIRE code was used for investigating the impact of multiply injection strategies and spray included angles on combustion and emissions in a marine diesel engine. The fuel injection parameters of spray included angle and pilot injection timing with pilot-main injection, as well as post injection ratio and post injection duration angle with pilot-main-post injection, were all investigated and optimized. The results indicate that retarding pilot injection timing with pilot-main injection declines high temperature region, resulting in a notable reduction in NOx emissions. Since fuel evaporation and burn are hampered by long spray penetration due to low temperature and pressure with pilot injection, a suitable spray included angle are used to offer more efficient air-fuel mixing process. A wider spray included angle simultaneously reduces soot emission and indicated specific fuel consumption (ISFC). Post injection fuel exerts impact on combustion process by causing a great disturbance to flow field during post combustion. Increasing post injection ratio from 4% to 10% at a small post injection duration angle great emission performance is achieved by simultaneous reduction in NOx and soot emissions while only using a slight consumption of ISFC. To summarize, the defeat of traditional NOx-soot trade-off occurs as both NOx and soot emissions are decreased with optimized multiple injection strategy and spray included angle. Particularly, there are respectively four cases with pilot-main injection and two cases with pilot-main-post injection, that achieve simultaneous reduction in NOx emissions, soot emission, and ISFC, compared to the prototype.

One month safety study of ExoFlo injection for the treatment of lumbar or cervical radiculopathy in the epidural space

This One month safety study addresses any potential risks behind the use of bone marrow-derived mesenchymal stem cell extracellular vesicle isolate product (ExoFlo Direct Biologics) as a treatment for cervical and lumbar radiculopathy. Ten healthy adults were treated with ExoFlo injections for cervical radiculopathy (n=5) and lumbar radiculopathy (n=5). Follow up occurred twenty-four hours, three days, one week, three weeks and one month post injection. By the one month follow-up, the average patient improved 55% in BPI, 55.2% in QD, 25.4% in UEFS, 19.75% in ODI and 26% in LEFS. There were no complications or adverse events by the end of the study and no patient exhibited worsening radiculopathy. Patients will continue to be followed for at least six months post injection.

VIRULENCE GENE PROFILING AND PATHOGENICITY OF Streptococcus agalactiae ISOLATED FROM TILAPIA, Oreochromis niloticus FARMS IN INDONESIA

Streptococcosis caused by Streptococcus agalactiae has become a major disease problem in tilapia culture in Indonesia. This study aimed to detect virulence genes of S. agalactiae during streptococcosis disease outbreaks in several tilapia farms in Indonesia and evaluate the correlation between biotype and virulence genes to bacterial virulence. The presence of virulence genes was determined in 10 strains of S. agalactiae isolated from farm-raised tilapia. Polymerase chain reaction (PCR) protocol was used to determine genes for cylE, hylB, lmb, bib A, PI-2b, fbs A, fbs B, gap, PI-1, and cfb in the template DNA. Pathogenicity test was carried out by intraperitoneal injection of 24 hour-cultured S. agalactiae to tilapia with 108 CFU/fish. Four isolates have seven of virulence genes (cylE, hylB, bibA, PI-2b, fbs A, fbs B, and gap genes), three isolates have six virulence genes (hylB, bib A, fbs A, fbs B, gap, cfb genes), one isolate has four virulence gene (hyl B, bib A, fbs, and cfb genes), and one isolate has one virulence gene (PI-2b gene). None of the isolates has lmb or PI-1 genes. Bacteria with more virulence genes showed higher pathogenicity post injection. Mortality of tilapia injected with b-hemolytic bacteria was 100% within the period of 14-19 hours, while non-hemolytic bacteria was 53.3%-86.6% on 14 days post-injection. Pathological changes associated with the infection by either isolate included melanosis, slow response, anorexia, ocular opacity, gasping, erratic, C-shape, and whirling. It can be concluded that S. agalactiae with more virulence genes show a higher level of pathogenicity. The presence of a virulent gene has the potential to be used as a basis for selecting candidate isolates and designing vaccine compositions as an effort to prevent streptococcosis infection in tilapia in Indonesia.

Osmotic Activity of Drugs is an Important Factor of Their Local Action at the Injection Site: What We Don't Use to Prevent Post-Injection Abscesses

It is established that very few modern high-quality drugs in the dosage form "Solution for injection" have isoosmotic activity in the range of 280-300 mosmol/l of water. The main part of solutions for injection is hypotonic or hypertonic solutions, which can have osmotic activity in the range of 0 - 4000 mosmol/l of water. The reason for this was that the osmotic activity of drugs is not included in the list of controlled indicators of drugs quality, so it is not subject to control. It is shown that the osmotic activity of drug solutions increases with an increase in the total concentration of all dissolved ingredients present in the solution. It was found that an excessively high concentration of the drug in the solution gives it an excessively large hyperosmotic activity, which can cause a local irritant and cauterizing effect at the injection sites due to dehydration of the cells. Therefore, to exclude post-injection abscesses, it is proposed to dilute the hypertonic drug with water for injection until it is given isotonic activity.

Nonclinical study and applicability of the absorbed dose conversion method with a single biodistribution measurement for targeted alpha-nuclide therapy

Background We recently reported a new absorbed dose conversion method, RAP (RAtio of Pharmacokinetics), for 211At-meta-astatobenzylguanidine (211At-MABG) using a single biodistribution measurement, the percent injected dose/g. However, there were some mathematical ambiguities in determining the optimal timing of a single measurement of the percent injected dose/g. Thus, we aimed to mathematically reconstruct the RAP method and to examine the optimal timing of a single measurement. Methods We derived a new formalism of the RAP dose conversion method at time t. In addition, we acquired a formula to determine the optimal timing of a single measurement of the percent injected dose/g, assuming the one-compartment model for biological clearance. Results We investigated the new formalism’s performance using a representative RAP coefficient with radioactive decay weighting. Dose conversions by representative RAP coefficients predicted the true [211At]MABG absorbed doses with an error of 10% or less. The inverses of the representative RAP coefficients plotted at 4 h post-injection, which was the optimal timing reported in the previous work, were very close to the new inverses of the RAP coefficients 4 h post-injection. Next, the behavior of the optimal timing was analyzed by radiolabeled compounds with physical half-lives of 7.2 h and 10 d on various biological clearance half-lives. Behavior maps of optimal timing showed a tendency to converge to a constant value as the biological clearance half-life of a target increased. The areas of optimal timing for both compounds within a 5% or 10% prediction error were distributed around the optimal timing when the biological clearance half-life of a target was equal to that of the reference. Finally, an example of RAP dose conversion was demonstrated for [211At]MABG. Conclusions The RAP dose conversion method renovated by the new formalism was able to estimate the [211At]MABG absorbed dose using a similar pharmacokinetics, such as [131I]MIBG. The present formalism revealed optimizing imaging time points on absorbed dose conversion between two radiopharmaceuticals. Further analysis and clinical data will be needed to elucidate the validity of a behavior map of the optimal timing of a single measurement for targeted alpha-nuclide therapy.