Novel Additive Solution "Aayusol" Significantly Preserves Platelets in Lesion-Free State during Extended Storage

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 3558-3558
Author(s):  
Jialan Shi ◽  
Xiu-Gui Lu ◽  
Hemant S Thatte

Abstract Introduction: We have recently demonstrated that hearts can be stored in a novel organ preservation solution Somah (Thatte HS et al, Circulation 2009), for an extended 24-hour period at ambient temperature, creating new paradigms in transplant surgery (Lowalekar SK et al, Transplant Proc 2013; Am J Transplant 2014). Somah was significantly modified to address the peculiarities of platelet biochemistry and metabolism, resulting in a novel platelet additive solution, Aayusol, that preserves platelets in a relatively lesion free state compared to PAS-IIIM during 9-day storage at ambient temperature. Methods: Platelets (20% PRP) were stored in transfer bags containing Aayusol or PAS-IIIM solution (20:80) at ambient temperature with gentle shaking under aseptic conditions. Immuno-Biochemical assays, flow cytometry, and semi-quantitative confocal microscopy were utilized to evaluate platelet viability and function (n=8) during 9-day storage. Results: The pH levels of both PASIIIM and Aayusol remained stable above 7.1 throughout 9-day storage. Significantly greater amount of platelet microparticles and aggregates were temporally formed, which peaked in PAS-IIIM (>8% & >4%) but not in Aayusol (<3% & <1.3%; *P < 0.05), at day 9 respectively (Figure 1 and Figure 2A and 2B). In contrast to PAS-IIIM platelets, Aayusol platelet morphology was well preserved during storage. Similarly, activation markers- PS exposure and P-selectin expression- (<15%) and (<5%) respectively, were significantly attenuated in Aayusol platelets, but not in PAS-IIIM at 9-day storage (Figure 2C and 2D). Furthermore, GPIba and GPIIb surface receptors, responsible for primary adhesion of circulating platelets to the thrombogenic surface, shed faster and decreased significantly in PAS-IIIM than in Aayusol at 9-day storage (GPIba: 30% vs. 60% and GPIIb: 40% vs. 60%; P < 0.05). Additionally, nitric oxide production was well preserved in Aayusol platelets, with NO fluorescence dye quantum yield of 190 units, which was significantly greater than the comparator, which yielded 105 units, at the end of storage (Figure 2E). As components in Aayusol specifically help modulate aerobic and anaerobic metabolism, lactic acid production was significantly lower in Aayusol (2.8±1.82 mmol/L) than in PAS-IIIM (3.9±0.32 mmol/L; P < 0.05). Correspondingly, high-energy phosphate synthesis (ATP+CP) was greater in Aayusol platelets compared to PAS-IIIM platelets. Temporal agonist (ADP and A23187) challenge resulted in greater activation of Aayusol platelets (over 70% PS and P-selectin expression) than PAS-IIIM (60% PS and 20% P-selectin; P < 0.05) on day 9 storage. Conclusion: Temporally assessed parameters showed superiority of Aayusol platelets compared to PAS-IIIM platelets. Therefore, based on conservation of structure and function, Aayusol platelet transfusion requirement will potentially be lower than PAS-IIIM platelets, thus increasing the availability of platelets to meet demand, with decreased costs and morbidity in patients. Thus, Aayusol may provide a more viable means of preserving platelets for transfusion. Disclosures No relevant conflicts of interest to declare.

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5110-5110
Author(s):  
Jialan Shi ◽  
Xiu-Gui Lu ◽  
Hemant S Thatte

Abstract Background: Extended storage of platelet in lesion free state is of critical importance and is tenuous at best in currently available additive solutions thus requiring innovations in storage modalities. We have recently demonstrated that donor hearts obtained for transplant can be stored at ambient temperature in Somah solution of our design. Unlike other clinically used solutions, rationally designed Somah maintains cellular homeostasis and high-energy state of the heart during extended storage, circumventing hypothermic injury and thus facilitating optimal functional recovery of heart upon reperfusion. This study was designed to evaluate the ability of Somah to preserve platelets in lesion free state during extended storage by modulation of analogous biochemical pathways as in the heart. Study Design and Methods: Blood was collected in ACD and the platelets (PRP) were isolated using standard protocol (n=3). Platelets were stored in Somah, PAS III and PAS III-M solutions at concentration of 65% at ambient temperature (22 oC) for 12 days under asepsis. The storage solutions were periodically assessed for pO2, pCO2, HCO-3, pH, Glucose and lactic acid using Abaxis iStat. Platelet morphology was evaluated with confocal microscopy; platelet activation and apoptosis was measure by quantitating and localizing phosphtidylserine (PS) exposure using Alexa 488-lactoadherin binding, flow cytometry, and confocal microscopy over the storage period. Results: Platelets demonstrated active metabolism in Somah. Glucose concentration decreased in Somah from 225 mg/dL to 110 mg/dL while the lactate concentration increased from 1.45 mMol/L to 9.0 mMol/L during the 12 day storage. Strong buffering capacity of Somah maintained the pH near neutrality. pO2 and pCO2remained at 165 mmHg and 30 mmHg respectively. In contrast, glucose decreased from 134 mg/dL to 102 mg/dL and lactate increased from 0.97 mMol/L to 15.6 mMol/L in the PAS group. PS exposure increased from 0.2% to 12.8 % at 12 days in platelets stored in Somah, in contrast, PS exposure increased from 0.2% to 30.7% during the same period in platelets stored in PAS solutions, respectively. Morphology of platelets was well maintained in Somah but not in the PAS solutions. There was no significance difference in measured values between PAS III and PAS III-M. Conclusion: Unique composition of Somah facilitates modulation of anaerobic and aerobic metabolism by diverting lactate into oxidative phosphorylation pathways, thus maintaining the energy state and cellular homeostasis. Intact morphology, active metabolism and minimal exposure of PS indicate that platelets were well preserved in Somah during extended storage. Work is under way to extend the storage further, evaluate high-energy stores, CD 62p, vWF expression and functional in vitro activation of platelets stored in Somah. These preliminary studies indicate Somah can be a viable alternative for storage of platelets in lesion free state during extended storage. Disclosures No relevant conflicts of interest to declare.


Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. SCI-1-SCI-1
Author(s):  
Douglas C. Wallace

Abstract Abstract SCI-1 The incidence and cost of the age-related metabolic and degenerative diseases is rising in the face of ever increasing investment. Therefore, something must be missing in our current hypotheses about these diseases. Until now, the prevailing paradigms in Western medicine have been the anatomical paradigm of disease which posits that tissue-specific symptoms are due to tissue-specific structural defects and the Mendelian paradigm of genetics which posits that if a disease is inherited according to the Laws of Mendel it is genetic and if not it is environmental. The epigenome is invoked to explain the environmental modulation of the nuclear gene expression. However, life requires both structure and energy and the over 1500 mitochondrial energy genes are dispersed across the chromosomes plus the maternally inherited mitochondrial DNA. Moreover, the cells and tissues most affected by aging and the age-related disease are those most reliant on mitochondrial energy and the mitochondrial lies at the interface between environmental calories and human physiology, diabetes and obesity. The epigenome and the signal transduction pathways are regulated by protein phosphorylation by ATP, acetylation via acetyl-CoA, and methylation by S-adenosylmethionine, all driven by mitochondrial high energy substrates modulated by available calories. Furthermore, mitochondrial redox chemistry regulates reactive oxygen production and thiol/disulfide chemistry and these also regulate cellular signaling and function. Therefore, bioenergetics and mitochondrial genetics are the missing factors which have inhibited our capacity of address the biology and genetics of the age-related metabolic and degenerative diseases, cancer and aging. Disclosures No relevant conflicts of interest to declare.


Author(s):  
Klaus-Ruediger Peters

Topographic ultra high resolution can now routinely be established on bulk samples in cold field emission scanning electron microscopy with a second generation of microscopes (FSEM) designed to provide 0.5 nm probe diameters. If such small probes are used for high magnification imaging, topographic contrast is so high that remarkably fine details can be imaged on 2DMSO/osmium-impregnated specimens at ribosome surfaces even without a metal coating. On TCH/osmium-impregnated specimens topographic resolution can be increased further if the SE-I imaging mode is applied. This requires that beam diameter and metal coating thickness be made smaller than the SE range of ~1 nm and background signal contributions be reduced. Subnanometer small probes can be obtained (only) at high accelerating voltages. Subnanometer thin continuous metal films can be produced under the following conditions: self-shadowing effect between metal atoms must be reduced through appropriate deposition techniques and surface mobility of metal atoms must be diminished through high energy sputtering and/or specimen cooling.


2010 ◽  
Vol 1 (1) ◽  
pp. 20-25 ◽  
Author(s):  
Michael Perry

Craniofacial trauma remains a common health problem throughout many areas of the UK. Although the ‘combination of alcohol and testosterone’ is often regarded as a major aetiological factor, a significant number of injuries are not related to either. Motor vehicle collisions and equine-related sports injuries in particular can result in devastating injuries to the skull and face and are frequently seen.Over the last few decades, management has moved away from closed methods to open exposure, anatomical reduction and internal ‘rigid’ fixation of facial fractures, with significant improvements in outcomes. Nevertheless, current management of ‘high energy’ or complex fractures can still result in residual functional disability and cosmetic deformity.Today’s challenge is to restore patients back to their pre-injury form and function, consistently, but this is not always possible. Greater understanding and a number of developments have significantly improved outcomes, although controversy still exists in some areas. Some of these will be discussed.


2018 ◽  
Vol 19 (10) ◽  
pp. 2907 ◽  
Author(s):  
Miyako Kurihara-Shimomura ◽  
Tomonori Sasahira ◽  
Chie Nakashima ◽  
Hiroki Kuniyasu ◽  
Hiroyuki Shimomura ◽  
...  

Head and neck cancers, including oral squamous cell carcinoma (OSCC), are the sixth most common malignancies worldwide. OSCC frequently leads to oral dysfunction, which worsens a patient’s quality of life. Moreover, its prognosis remains poor. Unlike normal cells, tumor cells preferentially metabolize glucose by aerobic glycolysis. Pyruvate kinase (PK) catalyzes the final step in glycolysis, and the transition from PKM1 to PKM2 is observed in many cancer cells. However, little is known about PKM expression and function in OSCC. In this study, we investigated the expression of PKM in OSCC specimens and performed a functional analysis of human OSCC cells. We found that the PKM2/PKM1 ratio was higher in OSCC cells than in adjacent normal mucosal cells and in samples obtained from dysplasia patients. Furthermore, PKM2 expression was strongly correlated with OSCC tumor progression on immunohistochemistry. PKM2 expression was higher during cell growth, invasion, and apoptosis in HSC3 cells, which show a high energy flow and whose metabolism depends on aerobic glycolysis and oxidative phosphorylation. PKM2 expression was also associated with the production of reactive oxygen species (ROS) and integration of glutamine into lactate. Our results suggested that PKM2 has a variety of tumor progressive functions in OSCC cells.


Author(s):  
Behrouz Shademan ◽  
Vahidreza Karamad ◽  
Alireza Nourazarian ◽  
Cigir Biray Avcı

Immunotherapy has become a prominent strategy for the treatment of cancer. A method that improves the immune system's ability to attack a tumor (Enhances antigen binding). Targeted killing of malignant cells by adoptive transfer of chimeric antigen receptor (CAR) T cells is a promising immunotherapy technique in the treatment of cancers. For this purpose, the patient's immune cells, with genetic engineering aid, are loaded with chimeric receptors that have particular antigen binding and activate cytotoxic T lymphocytes. That increases the effectiveness of immune cells and destroying cancer cells. This review discusses the basic structure and function of CAR-T cells and how antigenic targets are identified to treat different cancers and address the disadvantages of this treatment for cancer.


1998 ◽  
Vol 540 ◽  
Author(s):  
T. L. Daulton ◽  
R. S. Lewis ◽  
L. E. Rehn ◽  
M. A. Kirk

AbstractMetastable phase formation under highly non-equilibrium thermodynamic conditions within high-energy particle tracks are investigated. In particular, the possible formation of diamond by heavy-ion irradiation of graphite at ambient temperature is examined. This work was motivated, in part, by an earlier study which discovered nanometer-grain polycrystalline diamond aggregates of submicron-size in uranium-rich carbonaceous mineral assemblages of Precambrian age. It was proposed that these diamonds were formed within the particle tracks produced in the carbonaceous minerals by the radioactive decay of uranium. To test the hypothesis that nanodiamonds can form by ion irradiation, fine-grain polycrystalline graphite sheets were irradiated with 400 MeV Kr ions to low fluence (6 × 1012 ions-cm−2). The ion-irradiated (and unirradiated control) graphite were then subjected to acid dissolution treatments to remove the graphite and isolate any diamonds that were produced. These acid residues were characterized by transmission electron microscopy. The acid residue of the ion-irradiated graphite was found to contain nanodiamonds (at several ppm of bulk), demonstrating that ion irradiation of graphite at ambient temperature can produce diamond.


Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Ahmad Y Sheikh ◽  
Timothy C Doyle ◽  
Maryam K Sheikh ◽  
Feng Cao ◽  
Katherine J Ransohoff ◽  
...  

Background: Recent advances in high energy X-ray source computed tomography (CT) technology have made it possible to accurately image murine cardiac structure and function. We describe the use of a gated Micro CT system to assess cardiac remodeling in a murine model of myocardial infarction (MI). Methods: Adult FVB mice (n = 10) were randomized to surgically induced MI by left coronary artery ligation or sham procedure. Dual gated cardiac and respiratory Micro CT scans were performed pre-operatively and at weeks 4, 8 and 12 post-procedure. End-diastolic and end-systolic images were acquired by gating on the ECG P-and S’-waves, respectively. Post-acquisition analysis was performed using image analysis software and the following parameters were quantified: left ventricular (LV) volume and mass, right ventricular volume and mass, right and left atrial volumes, and ventricular ejection fraction (EF). Results: Images were successfully acquired with a resolution of 100 microns allowing for identification and quantification of key cardiac structures (Fig 1A ). Following MI, animals exhibited left ventricular failure with significantly increased* end systolic and diastolic volumes by week 4. Ventricular dilation continued through week 8, plateauing by week 12. Left ventricular mass increased steadily over 12 weeks*, with a significantly decreased* LVEF of 28.0 ± .05% by week 12 (pre-MI: 66.7 ± .06%,*p < 0.01). Post-MI left ventricular change is detailed in Fig 1B . Conclusions: MicroCT scanning can be successfully used to characterize murine myocardial structure and function, making it a useful tool to assess cardiac phenotypes and models of cardiovascular disease.


Cancers ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 1783 ◽  
Author(s):  
Ping-Hsiu Wu ◽  
Abayomi Emmanuel Opadele ◽  
Yasuhito Onodera ◽  
Jin-Min Nam

Due to advancements in nanotechnology, the application of nanosized materials (nanomaterials) in cancer diagnostics and therapeutics has become a leading area in cancer research. The decoration of nanomaterial surfaces with biological ligands is a major strategy for directing the actions of nanomaterials specifically to cancer cells. These ligands can bind to specific receptors on the cell surface and enable nanomaterials to actively target cancer cells. Integrins are one of the cell surface receptors that regulate the communication between cells and their microenvironment. Several integrins are overexpressed in many types of cancer cells and the tumor microvasculature and function in the mediation of various cellular events. Therefore, the surface modification of nanomaterials with integrin-specific ligands not only increases their binding affinity to cancer cells but also enhances the cellular uptake of nanomaterials through the intracellular trafficking of integrins. Moreover, the integrin-specific ligands themselves interfere with cancer migration and invasion by interacting with integrins, and this finding provides a novel direction for new treatment approaches in cancer nanomedicine. This article reviews the integrin-specific ligands that have been used in cancer nanomedicine and provides an overview of the recent progress in cancer diagnostics and therapeutic strategies involving the use of integrin-targeted nanomaterials.


Sign in / Sign up

Export Citation Format

Share Document