Short-Term Storability of Alginate-Encapsulated Persian Violet Microshoots for Germplasm Exchange

Microshoots have been widely used for micropropagation. It may be necessary to store microshoots for a short period of time, for example in germplasm exchange needing transport to other research groups. Here, we investigated the short-term storability of alginate-encapsulated Persian violet (Exacum affine Balf. f. ex Regel) microshoots at 4 °C and 25 °C. After storage, the encapsulated microshoots were sown on basal Murashige and Skoog medium for germination and viability determination using tetrazolium chloride staining. The results showed that one or five microshoots encapsulated with a single alginate layer could be stored at 4 °C for up to 30 days, while the percentages of germination and viability of the microshoots encapsulated with two layers of alginate were greatly reduced upon storage. This is the first report on the storability of alginate-encapsulated multiple microshoots, which could be a more efficient way to encapsulate microshoots used for short-term cold storage.

GSK3β Serine 389 Phosphorylation Modulates Cardiomyocyte Hypertrophy and Ischemic Injury

Prior studies show that glycogen synthase kinase 3β (GSK3β) contributes to cardiac ischemic injury and cardiac hypertrophy. GSK3β is constitutionally active and phosphorylation of GSK3β at serine 9 (S9) inactivates the kinase and promotes cellular growth. GSK3β is also phosphorylated at serine 389 (S389), but the significance of this phosphorylation in the heart is not known. We analyzed GSK3β S389 phosphorylation in diseased hearts and utilized overexpression of GSK3β carrying ser→ala mutations at S9 (S9A) and S389 (S389A) to study the biological function of constitutively active GSK3β in primary cardiomyocytes. We found that phosphorylation of GSK3β at S389 was increased in left ventricular samples from patients with dilated cardiomyopathy and ischemic cardiomyopathy, and in hearts of mice subjected to thoracic aortic constriction. Overexpression of either GSK3β S9A or S389A reduced the viability of cardiomyocytes subjected to hypoxia–reoxygenation. Overexpression of double GSK3β mutant (S9A/S389A) further reduced cardiomyocyte viability. Determination of protein synthesis showed that overexpression of GSK3β S389A or GSK3β S9A/S389A increased both basal and agonist-induced cardiomyocyte growth. Mechanistically, GSK3β S389A mutation was associated with activation of mTOR complex 1 signaling. In conclusion, our data suggest that phosphorylation of GSK3β at S389 enhances cardiomyocyte survival and protects from cardiomyocyte hypertrophy.

Express procedure of intraoperative estimation of viability of small bowel in experiment and clinic

Objective. Elaboration of effective and informative method of the small intestine viability determination for improvement of the surgical treatment results in pathologies, which need performance of resection of its segments. Materials and methods. The work is considered as experimentally-clinic one. Its experimental part was conducted on the rats; temporary ischemia was simulated, using application of tourniquet on the intestinal loop, taken from abdominal cavity after doing laparotomy; the animals were distributed on the groups, depending on the ischemia duration applied. Clinical part of the investigation was performed on 68 patients, including a control group of 50 patients, in whom intestinal resection was done, taking into account subjective estimation of the gut viability, and the main group, consisted of 18 patients, in whom intestinal resection was performed, basing on the gut viability estimation data, obtained using the method elaborated. Results. In the animals, on which temporary noncritical ischemia (up to 10 min) was simulated, the ischemia zone did not differed visually from other intestinal segments, while after persistence of ischemia up to 20- and 25-min the changes have had appeared necrotic with presence of peritonitis. Studying of intestinal specimen in the animals, in whom an acute ischemia was simulated, have shown the mostly expressed changes in endotheliocytes of microcirculatory bed, depending on duration of the ischemia dystrophic changes have been enhanced up to destructive. Total electric resistance (іmpedance) was measured on various frequencies. On all the frequencies, the raising of total electric resistance with more durable ischemia period was observed. Conclusion. Changes of total electric resistance are reflecting the degree of the blood flow disorder in intestinal wall and may be exploited as a criterion of determination of its viability (capacity of the anastomosis to heal). Objectivization of estimation of the intestinal wall state reduces the quantity of resection of injured intestinal segment and the morbidity.

F18-FDG Cardiac PET/CT: An alternative tool for myocardial viability determination prior to coronary revascularization decision in severe ventricular dysfunction

Purposes: The aim of the present study was to evaluate clinical value and accuracy of Fludeoxyglucose Cardiac Positron Emission Tomography Computerized Tomography (FDG C-PET/CT) as an alternative tool for myocardial viability determination prior to coronary revascularization decision in lower left ventricular ejection fraction (LVEF) patients. We accepted lower LVEF as 35% or less for the study. FDG C-PET/CT is reported to be the gold standart for myocardial viability detection among other techniques such as thallium-201 rest-redistribution scintigraphy myocardial perfusion imaging and dobutamine stress echocardiography. Materials and Methods: Between the dates of 01.01.2010 and 10.07.2019, 191 consecutive patients (mean age 64±9.1 years) underwent CABG operations with severe LVEF dysfunction with 35% or less. These impaired LVEF patients were calculated as the 4.4% total CABG cases. Myocardial viability was also studied and elaborated by F18-FDG C-PET/CT for all cases. Elaboration was detailed as viable myocardial tissue, hibernated myocardial tissue or necrosis areas (scars, nonviable tissue) via segmentally images of the heart. Final surgical decision was primarily depended on F18-FDG C-PET/CT for the majority of cases. Results: 191 CABG operations were performed. Perioperative deaths occured in 18 (9.4%) cases. Mortality reasons were prolonged CPB in 4 (2%) cases, severe and unmanageable ventricular arrhytmias in 5 (2.6%) cases, MOD with prolonged intubation in 6 (3.1%) cases and major neurological complication in 2 (1%) cases. Our mean coronary graft number was 3.5±0.8. LIMA was used in the majority of cases (n=176, 92.1%). LIMA was anastomosed to LAD for each case. IABP insertions were applied in 81(42.4%, 43 cases intraoperatively, 38 cases at ICU) patients. 236 patients with impaired LVEF and severe coronary artery disease were evaluated by F18-FDG C-PET/CT prior to operation. 191 cases (80.9%) were accepted as canditates for revascularization with multiple viable segments.45 cases (19.1%) presented transmural scar tissue (non-viable) images by FDG uptake analysis. This group cases were considered to be with non-beneficial results from revascularization. Thus, these patients were referred to medical treatments. Mean number of viable segments on F18-FDG C-PET/CT were calculated as 5.2±1.4 for each patient. Conclusions: The presence of myocardial viability is crucial to define reasonable canditates for revascularization in cases with lower LVEF. Among other preoperative viability detection techniques such as dobutamin echocardiography and myocardial perfusion scintigraphies, F18 FDG cardiac PET/CT is accepted as the ‘Gold Standart’ for segmental analysis on basis of distinguishing scar tissue from viable components. We strongly advocate F18-FDG cardiac PET/CT to be a suitable and effective tool to evaluate CABG indications in severe ventricular dysfunction.

Viability determination of Ascaris ova in raw wastewater: a comparative evaluation of culture-based, BacLight Live/Dead staining and PMA-qPCR methods

Accurate evaluation of viable Ascaris ova in wastewater is the key to mitigating Ascaris reinfections in endemic regions. In this study, the viability of Ascaris ova in raw wastewater was determined using three different detection methods: culture-based, BacLight Live/Dead staining and propidium monoazide–quantitative polymerase chain reaction (PMA-qPCR). Furthermore, comparative assessment of viability utilising the aforementioned detection methods was performed using seeded experiments in wastewater. The percentage of viability was: culture-based (82%), BacLight Live/Dead staining (87%) and PMA-qPCR (85%) respectively. Despite the fact that no statistical difference was shown in the viability determination among the three methods, PMA-qPCR-based viability determination would be preferable over the other two methods for evaluating potential public health risks with A. suum ova due to its accuracy, being least subjective and its rapid reaction time.

Ability of Vital and Fluorescent Staining in the Differentiation of Schistosoma haematobium Live and Dead Eggs

This study reports (for the first time) the staining ability of vital (0.4% trypan blue and 1% neutral red) and fluorescent (Hoechst 33258) dyes to differentiate between live and dead Schistosoma haematobium (S. haematobium) eggs in human urine samples. Since S. haematobium egg is important in disease pathology, diagnosis, transmission, and drug development research, it is essential to be able to easily distinguish live eggs from dead ones. Staining is considered a way of enhancing the identification of live and dead eggs. Urine samples from school children were examined for the presence of S. haematobium eggs. Vital and fluorescent dyes were used to stain the samples that contained S. haematobium eggs, after which they were observed using light and fluorescent microscopes, respectively. The Hoechst 33258 provided a good staining outcome for differentiation between live and dead eggs, followed by 0.4% Trypan blue. Regarding the 1% neutral red stain, even though it provided some evidence of which egg was alive or dead, the distinction was not very clear; therefore, it could be useful when used in combination with other stains for egg viability determination. The benefits of this study will include assessing the effect of drugs on S. haematobium eggs in Schistosomiasis research.