Overnight Storage Research Articles

Impact of Overnight Storage of Human Atrial Myocytes on Intracellular Calcium Homeostasis and Electrophysiological Utility.

Human atrial myocytes afford an attractive experimental model to investigate mechanisms underlying electrophysiological alterations in cardiovascular disease. However, this model presents limitations, such as the availability of human atrial tissue and a variable yield of myocytes isolation. Therefore, we aimed to determine whether overnight storage can increase the time window where the electrophysiological properties of human atrial myocytes can be determined. To address this issue, human atrial myocytes isolated from patients undergoing cardiac surgery were used for patch-clamp experiments on the day of cell isolation (Day 1) and the following day (Day 2). The shape of the current-voltage (I-V) relationship for the calcium current (ICa) depended on the access resistance and the cell capacitance, with large cells (>75 pF) requiring a lower access resistance (<15 MΩ) than small cells (<40 pF) to avoid distortion of the I-V curve. Importantly, overnight storage did not significantly affect (1) the ICa amplitude or properties, (2) sarcoplasmic reticulum calcium homeostasis or (3) the frequency-dependency of the beat-to-beat response. In conclusion, overnight storage of isolated human atrial myocytes at 4 °C does not affect essential features of intracellular calcium homeostasis and, therefore, affords a simple protocol to extend the experimental lifetime of human atrial myocytes.

Proteomic Profiles of Human Arterioles Isolated From Fresh Adipose Tissue or Following Overnight Storage

Arterioles are key determinants of the total peripheral vascular resistance, which, in turn, is a key determinant of arterial blood pressure. However, the amount of protein available from one isolated human arteriole may be less than 5 μg, making proteomic analysis challenging. In addition, obtaining human arterioles requires manual dissection of unfrozen clinical specimens. This limits its feasibility, especially for powerful multicenter clinical studies in which clinical specimens need to be shipped overnight to a research laboratory for arteriole isolation. We performed a study to address low-input, test overnight tissue storage and develop a reference human arteriolar proteomic profile. In tandem mass tag proteomics, use of a booster channel consisting of human induced pluripotent stem cell–derived endothelial and vascular smooth muscle cells (1:5 ratio) increased the number of proteins detected in a human arteriole segment with a false discovery rate of <0.01 from 1051 to more than 3000. The correlation coefficient of proteomic profile was similar between replicate arterioles isolated freshly, following cold storage, or before and after the cold storage (1-way analysis of variance; P = .60). We built a human arteriolar proteomic profile consisting of 3832 proteins based on the analysis of 12 arteriole samples from 3 subjects. Of 1945 blood pressure–relevant proteins that we curated, 476 (12.5%) were detected in the arteriolar proteome, which was a significant overrepresentation (χ2 test; P < .05). These findings demonstrate that proteomic analysis is feasible with arterioles isolated from human adipose tissue following cold overnight storage and provide a reference human arteriolar proteome profile highly valuable for studies of arteriole-related traits.

Fluid and Thermal Analysis of Pre-Columbian Tiwanaku (500–1100 CE) Raised-Field Agricultural Systems of Bolivia

Raised-field agricultural systems have received attention from scholars involved in the analysis of prehistoric agricultural intensification in the New World. This paper discusses the function of raised fields associated with the Tiwanaku society (500–1100 CE) located on the southern rim of Lake Titicaca in Bolivia. The overnight internal heat storage capacity of Tiwanaku raised-field berms located at the high-altitude (~3810 masl) Bolivian altiplano is analyzed through ANSYS (version 4.2B) finite difference methods to provide an understanding of ancient agricultural engineers’ knowledge regarding how to protect crops from nightly subzero freezing temperatures and water saturation. The present analysis concludes that enhanced berm heat storage capacity derived from solar radiation into multi-layered moist berm agricultural soils, together with radiative heating of berm-surrounding swale water (swale water depth determined from excavation into the groundwater aquifer), was an essential Tiwanaku design element of raised-field agriculture to protect crops from freezing damage during both wet and dry seasons. This paper reports the ANSYS temperature distribution results derived from a raised-field berm swale computer model of ancient excavated raised fields in the form of a 24 h heat input and cooling cycle, which indicates the presence of an internal berm heat storage effect designed to protect crops from freezing damage. The calculations performed use specific hydrological and climatological conditions characteristic of the littoral and near-shore environment of Lake Titicaca. The use of the ANSYS finite element code to investigate the source of internal berm heat storage protecting crops from freezing temperatures, compared to the field test results from experimental use of reconstructed ancient, raised fields, provides an understanding of the technologies developed by Tiwanaku agricultural engineers to increase raised-field agricultural production.

Abstract 132: Proteomic Profiles Of Human Arterioles Isolated From Fresh Adipose Tissue Or Following Overnight Storage

Resistance arteries, or arterioles, are key determinants of the total peripheral vascular resistance, which, in turn, is a key determinant of arterial blood pressure (BP). However, the amount of protein available from one isolated human arteriole may be less than 5 μg, making proteomic analysis challenging. In addition, obtaining human arterioles requires manual dissection of unfrozen clinical specimens. This limits its feasibility, especially for powerful multi-center clinical studies in which clinical specimens need to be shipped overnight to a research lab for arteriole isolation. We performed a study to address low input, test overnight tissue storage, and develop a reference human arteriolar proteomic profile. We found that, in tandem mass tag proteomic analysis, the use of a booster channel consisting of endothelial and vascular smooth muscle cells (1:5 ratio) increased the number of proteins detected in a human arteriole segment with FDR &lt; 0.01 from 1,051 to more than 3,000. We collected adipose tissues from three human subjects and isolated two arterioles from each fresh aliquot of the tissue or after 24h of cold storage of unfrozen aliquots in MACS tissue solution. The correlation coefficient of proteomic profile was similar (p=0.6) between replicate arterioles isolated freshly, following the cold storage, or before and after the cold storage. We built a human arteriolar proteomic profile consisting of 3,836 proteins based on the analysis of 12 arteriole samples from the three subjects. The average arteriolar protein had a mean copy number of 2.76х10 6 per cell using the histone proteomic ruler, which is based on the fact that the mass of DNA per cell is approximately equal to the protein mass of histones. Transgelin was the most abundant protein detected. We curated a set of BP-relevant human genes, which encode 1,945 proteins. Of these BP-relevant proteins, 476 (12.5%) were detected in the arteriolar proteome, which was a significant overrepresentation (p&lt;0.05, Chi squared test). These findings demonstrate that proteomic analysis is feasible with arterioles isolated from human adipose tissue following cold overnight storage and provide a reference human arteriolar proteome profile highly valuable for studies of arteriole-related traits.

Subzero saline chilling with or without prechilling in icy water improved chilling efficiency and meat tenderness of broiler carcasses

Freshly slaughtered carcasses need to be chilled to improve product quality, meat safety, and processing efficiency. This research investigated the effect of subzero saline chilling (SSC) on broiler carcasses with or without prechilling in icy water. Water immersion chilling at 0.5°C (WIC) or SSC at 4% NaCl/-2.41°C (SSC) was a major chilling step. For the combination of pre- and postchilling, the warm water immersion chilling (WWIC) at 10°C was used as prechilling and the WIC as postchilling (WWIC-WIC), and WIC was used as prechilling and the SSC as postchilling (WIC-SSC). The internal temperature of breast fillets was monitored during chilling. Carcasses in a prechiller were transported to a postchiller when their internal temperature reached 15°C. Chilling was completed when the carcass temperature reached 4.4°C or below, and breast fillets were harvested at 3-h postmortem to measure the pH and sarcomere length. Color (L*, a*, and b*) values were evaluated on both breast skin and skinless breast surfaces. Meat tenderness was evaluated using the breast fillets after overnight storage and cooking to an internal temperature of 76°C. The carcasses in the SSC and WIC-SSC showed shorter chilling times (85-91 min) than those (100-144 min) of WIC and WWIC-WIC. A higher chilling yield was observed for the carcasses in WIC-SSC, and a lower cooking yield was seen for the carcasses in WWIC-WIC than other chilling methods (P < 0.05). The breast fillets of broilers in the SSC and WIC-SSC showed lower shear forces and longer sarcomere length than the WIC and WWIC-WIC. No difference was found for L* and a* values, while lower b* value was observed in the SSC than the other chilling methods (P < 0.05). Based on these results, chilling of broiler carcasses in the SSC (4% NaCl/-2.41°C) with or without prechilling in WIC at 0.5°C significantly improved chilling efficiency and meat tenderness, with minor color changes on carcasses.

Low internal air space in plants with crassulacean acid metabolism may be an anatomical spandrel.

Crassulacean acid metabolism (CAM) is a photosynthetic adaptation found in at least 38 plant families. Typically, the anatomy of CAM plants is characterized by large photosynthetic cells and a low percentage of leaf volume consisting of internal air space (% IAS). It has been suggested that reduced mesophyll conductance (gm) arising from low % IAS benefits CAM plants by preventing the movement of CO2 out of cells and ultimately minimizing leakage of CO2 from leaves into the atmosphere during day-time decarboxylation. Here, we propose that low % IAS does not provide any adaptive benefit to CAM plants, because stomatal closure during phase III of CAM will result in internal concentrations of CO2 becoming saturated, meaning low gm will not have any meaningful impact on the flux of gases within leaves. We suggest that low % IAS is more likely an indirect consequence of maximizing the cellular volume within a leaf, to provide space for the overnight storage of malic acid during the CAM cycle.

Filter sterilized carcass rinsate for recovery of Salmonella species with various concentrations of cetylpyridinium chloride.

Controlling Salmonella in poultry processing continues to be important to processors and consumers. Cetylpyridinium chloride (CPC) has proven to be effective in vitro in controlling Salmonella. This study evaluated the recovery of Salmonella after overnight storage in 4°C filter-sterilized carcass rinsate containing CPC from 0.44 to 909 ppm (μg/mL). Ten Salmonella serotypes (18 strains), of which 6 serotypes are commonly isolated from poultry products, were grown in Bacto-Tryptic Soy Broth overnight at 37°C. Serial dilutions of a CPC/propylene glycol solution were prepared in 24-well tissue culture plates containing filter-sterilized carcass rinsate. Approximately 107 cfu/mL of each Salmonella serotype was added to the appropriate wells. Inoculated plates were stored overnight at 4°C. After storage, triplicate plates of brilliant green agar with sulfapyridine (BGS) were surface inoculated with 10 μL of the contents for each well, streaked for isolation, and incubated at 37°C for 24 h. Three replications were conducted. The presence of typical colonies on BGS plates was recorded as growth and verified through biochemical and serological testing. Of the serotypes chosen, Salmonella Kentucky, Dublin, and Enteritidis were the least resistant to CPC with a median minimum inhibitory concentration (MIC) of 14.22 μg/mL (range from 3.55 to 56.88 μg/mL); S. Typhimurium demonstrated a median MIC of 114.00 μg/mL (range from 28.44 to 114.00 μg/mL). Residual CPC potentially remaining attached to a carcass or in the weep after processing could potentially alter which Salmonella serotype is recovered from a carcass rinse due to different growth patterns during regulatory testing, with a potential for more virulent strains not to be recovered.

Hot or not? Investigating radiation safety and policy updating for the appropriate storage of radioactive specimens at Mount Sinai Hospital

PurposeThe current hospital policy for this study is stringent regarding the storage of radioactive sentinel lymph node (SLN) specimens, which requires the storage time of 24 hours before being handled by Pathology. Additional labeling along with separate containment of these specimens can be forgone if negligible radiation levels are found. The objective of this study was to determine whether the storage time needed for resected radioactive breast and primary site specimens to decay to twice the background radiation levels is less than 24 hours. MethodsThe investigators retrieved breast and primary site SLN specimens from the Pathology department on the same day of the biopsy. A dose calibrator was used to measure the dose, specimen, and concurrent background radioactivity in Megabecquerels (MBq). Radioactive decay calculations were used to further investigate when specimen activities reached twice the background levels. A retrospective analysis was performed using a one-sample t-test to determine if the time to reach double the background activity was significantly different from 24 hours. This study pertained to workflow optimization; thus, general procedure consent forms were sufficient. Both patient confidentiality and privacy were protected. The investigators followed the necessary radiation safety measures. ResultsThe mean time for specimens to reach twice the background level of radioactivity was 3.99 hours, significantly less than current storage time of 24 hours (p < 0.001). The mean time point for the SLNs to reach 1/16th of the original activity was 7.78 hours (p < 0.001). The average node activity was 0.14 MBq. ConclusionThe average sentinel node activity was less than 1 exemption quantity and the time to reach less than twice the background levels was significantly less than 24 hours, meaning that radioactive labels are not needed, and the 24-hour overnight specimen storage can be mitigated.

Optimisation of Curcumin Microemulsions Using Palm Oil

Curcumin is a natural hydrophobic polyphenol derived from the curcuminoids of Curcuma longa. Curcumin is commonly known as turmeric and it gains the interest of scientific and clinical researchers as it exhibits great pharmacological benefits such as anti-cancer, antiinflammatory and anti-oxidant properties. Nevertheless, curcumin is still not an approved drug in clinical settings due to its poor aqueous solubility and low oral bioavailability. Therefore, a self-microemulsifying drug delivery system (SMEDDS) was used as an approach to enhance the solubility and bioavailability of curcumin. The microemulsion was devised in a pre-formulation phase using a surfactant (Tween 80), a co-surfactant (polyethylene glycol, PEG 400), a lipid phase (palm oil) and an aqueous phase (water). A ternary phase diagram was used to identify the self-microemulsifying region in a formulation. Five of these formulations (F1, F2, F4, F7 and F10) were found to be stable with no phase separation observed upon overnight storage. All of the five formulations (except F4) possessed a high percentage of transmittance (86%–100%), which signified the formation of a stable microemulsion when they were diluted in a 1:100 ratio by water. Curcumin microemulsions were formulated by loading curcumin into F1, F2, F7 and F10. Only F1 and F2 curcumin microemulsions exhibit a clear appearance, however, F7 and F10 form a turbid solution, which indicates the formation of the emulsion. The results indicated that F1 and F2 which contain a high surfactant/co-surfactant-to-oil ratio of 9:1 is optimum to formulate the curcumin microemulsions.

Characterization of polyvalent Escherichia phage Sa157lw for the biocontrol potential of Salmonella Typhimurium and Escherichia coli O157:H7 on contaminated mung bean seeds.

Seeds are one of the primary sources of contamination with foodborne pathogens, such as pathogenic Escherichia coli, and various Salmonella serovars, for produce, particularly sprouts. Due to the susceptibility of sprout growth to chemical-based antimicrobials and the rising issue of antimicrobial resistance, developing innovative antimicrobial interventions is an urgent need. Therefore, the objective of this study was to characterize Escherichia phage Sa157lw (or Sa157lw) for the biocontrol potential of Salmonella Typhimurium and E. coli O157:H7 on contaminated mung bean seeds. Phage Sa157lw was subjected to whole-genome sequencing and biological characterization, including morphology, one-step growth curve, and stress stability tests. Later, antimicrobial activity was determined in vitro and upon application on the mung bean seeds artificially contaminated with E. coli O157:H7 or Salmonella Typhimurium. Sa157lw possessed a contractile tail and belonged to the Kuttervirus genus under the Ackermannviridae family, sharing a close evolutionary relationship with E. coli phage ECML-4 and Kuttervirus ViI; however, tail spike genes (ORF_102 and ORF_104) were the primary region of difference. Comparative genomics showed that Sa157lw encoded a cluster of tail spike genes-including ORF_101, ORF_102, and ORF_104-sharing high amino acid similarity with the counterfeits of various Salmonella phages. Additionally, Sa157lw harbored a unique tail fiber (ORF_103), possibly related to the receptors binding of O157 strains. The genomic evidence accounted for the polyvalent effects of Sa157lw against E. coli O157:H7 and various Salmonella serovars (Typhimurium, Enteritidis, Agona, Saintpaul, and Heidelberg). Furthermore, the phage did not contain any virulence, antibiotic-resistant, or lysogenic genes. Sa157lw had a 30-min latent period on both E. coli O157:H7 and Salmonella Typhimurium, with an estimated burst size of 130 and 220 PFU/CFU, respectively, and was stable at a wide range of temperatures (4-60°C) and pH (pH4 to pH10). The phage application demonstrated a strong anti-E. coli O157:H7 and anti-Salmonella Typhimurium effects in 1.1 and 1.8 log reduction on the contaminated mung bean seeds after overnight storage at 22°C. These findings provide valuable insights into the polyvalent Sa157lw as a potential biocontrol agent of Salmonella Typhimurium and E. coli O157:H7 on sprout seeds.

Effect of nucleated cell count and cryopreservation on engraftment post autologous stem cell transplant

Using 753 collections from 426 adult haematology patients, we conducted a retrospective, analysis into the effects of overnight storage and nucleated cell counts (NCC) on viable, CD34+ (vCD34+) recovery and engraftment kinetics post autologous stem cell, transplant (ASCT) with peripheral blood stem cells (PBSC). There were significant, differences in vCD34 + recovery ( P < 0.01) after cryopreservation associated with, the fresh NCC of ≥ 300 × 10 6 /mL in products stored overnight, but no association, with time to platelet or neutrophil engraftment post-ASCT was observed for these, products. There was no association of vCD34+ numbers or engraftment kinetics with cryopreserved NCC with either below or greater than the local recommended concentration of 400 × 106 /mL of product. However, there was significant difference in engraftment kinetics in relation to the viable CD34+ dose given at ASCT, in relation to the time to early engraftment and the amount of platelet support given during the engraftment period post-ASCT. We conclude the vCD34+ dose at ASCT is of great importance to early engraftment kinetics and that NCC is an important factor during overnight storage, but not for cryopreservation of PBSC. In light of our findings, we recommend that apheresis products collected in a closed system can safely be stored undiluted overnight.

Measurement of Tetrahydrobiopterin in Animal Tissue Samples by HPLC with Electrochemical Detection-Protocol Optimization and Pitfalls.

Tetrahydrobiopterin (BH4) is an essential cofactor of all nitric oxide synthase isoforms, thus determination of BH4 levels can provide important mechanistic insight into diseases. We established a protocol for high-performance liquid chromatography/electrochemical detection (HPLC/ECD)-based determination of BH4 in tissue samples. We first determined the optimal storage and work-up conditions for authentic BH4 and its oxidation product dihydrobiopterin (BH2) under various conditions (pH, temperature, presence of antioxidants, metal chelators, and storage time). We then applied optimized protocols for detection of BH4 in tissues of septic (induced by lipopolysaccharide [LPS]) rats. BH4 standards in HCl are stabilized by addition of 1,4-dithioerythritol (DTE) and diethylenetriaminepentaacetic acid (DTPA), while HCl was sufficient for BH2 standard stabilization. Overnight storage of BH4 standard solutions at room temperature in HCl without antioxidants caused complete loss of BH4 and the formation of BH2. We further optimized the protocol to separate ascorbate and the BH4 tissue sample and found a significant increase in BH4 in the heart and kidney as well as higher BH4 levels by trend in the brain of septic rats compared to control rats. These findings correspond to reports on augmented nitric oxide and BH4 levels in both animals and patients with septic shock.

Clinical Issues—April 2022

Perioperative care of the patient with alpha-gal syndrome Key words: alpha-gal syndrome (AGS), red meat allergy, allergic reaction, mammalian-derived ingredients, gelatin. Overnight storage of sterile supplies Key words: sterile supplies, event-related sterility, terminal cleaning, damp dusting, instrument table. Unsterile medication cartridges Key words: dental cartridge, local anesthesia, lidocaine, alcohol, sterile field. Personnel traffic into the OR Key words: OR traffic, door openings, airflow, environmental contamination, automatic doors.

Immobilization of naringinase on asymmetric organic membranes: Application for debittering of grapefruit juice

An enzymatic membrane reactor (EMR) is assembled for the immobilization of naringinase on a polyethersulfone ultrafiltration membrane, based on fouling-induced method. The effects of molecular weight cut-off, membrane configuration, applied pressure, enzyme concentration and pH are studied in terms of permeate rate, immobilization efficiency, and biocatalytic conversion. The 10 kDa membrane operating in reverse mode, 0.2 MPa, 0.3 gL−1 of enzyme in acetate buffer at pH 5 and cross-linking with 0.25% glutaraldehyde showed the highest naringin conversion (73%). It was determined that the intermediate pore blocking model was the predominant fouling mechanism for the enzymatic immobilization. The EMR was applied for debittering of grapefruit juice, achieving a conversion of naringin below bitterness threshold and maintaining the antioxidant capacity of the juice. Furthermore, the biocatalytic activity of immobilized enzyme was retained at a high level at least during three consecutive reaction runs, with overnight storage at 4 °C after each run. Industrial relevanceThe potential of membrane technologies in the juice industries is widely recognized today. The development of EMR with naringinase activity is an attractive option for reducing bitterness that could replace current techniques, due to its high specificity and effectiveness, possibility of repeated and continuous use, and in order to retain the properties of juice as much as possible. The research therefore represents an advance in the application of biocatalytic membranes as technological alternative for juice debittering.

Effects of overnight storage conditions on conventional complete removable prostheses.

To evaluate the effects of overnight storage conditions on the dimensional stability and retention of prostheses, as well as the participant's subjective perception of these interventions. Subjects with maxillary complete prostheses (CRPs) were randomly assigned to receive two interventions in a crossover design: storing the CRPs overnight in a dry or wet environment with a cleansing tablet. The denture intaglio surface was scanned, and outcome measures were collected before each intervention (BLN), post ntervention (PIS), and after immersion in water for 15 minutes after the intervention (WOC). Dimensional changes were analyzed for the total surface, anterior flange, palate, and tuberosities. Retention force was measured using a dynamometer, and the participants' subjective assessment of comfort, fit, and retention of their CRPs were collected. After verifying normal distribution, paired t and Wilcoxon signed-rank tests were used to check for any statistical significance (α = .05). Ten participants (mean age: 76.5 ± 5.9 y) were recruited in this study. Between BLN and PIS, the dimensional changes after dry intervention were significantly less than when stored wet for total surface (P = .009), anterior flange (P = .028), and the palate (P = .005). No difference was found between the effects of storage or washout interventions on objectively measured CRP retention. However, after WOC following dry storage, participants perceived a lower retention (P = .021), and a more comfortable palate after WOC following the wet intervention (P = .018). For dimensional stability, dry overnight storage of removable prostheses can safely be recommended when indicated. Immersion in water for 15 minutes does not seem to add advantages.

Development and Validation of an Analytical Method for Quantitation of Alpha-pinene in Rodent Blood and Mammary Gland by Headspace GC-MS.

Alpha-pinene (AP), produced by pine trees and other plants, is the main component of turpentine and is used as a fragrance and flavor ingredient. Exposure occurs via use of personal care and household cleaning products and in the lumber industry. Despite widespread exposure, toxicity data for AP are limited. The objective of this work was to develop and validate a method to quantitate AP in rodent blood and mammary glands, in support of toxicokinetic and toxicology studies of AP. The method uses 100 µL of blood or ~100 mg of mammary gland with analysis by headspace gas chromatography-mass spectrometry. The samples are diluted with internal standard (2H3-AP, IS) and sealed in headspace vials; mammary glands are homogenized within the vial. The vials are equilibrated briefly at 60°C before a headspace sample is analyzed. The method was validated in Sprague Dawley rat blood over the range 5-500 ng/mL and mammary gland over the range 100-5000 ng/g. The method was linear (r ≥0.99), accurate (mean relative error (RE) ≤±13.4%) and precise (relative standard deviation (RSD) ≤7.1%) in both matrices. Recoveries incorporating IS were ≥88.7% at all concentrations in both tissues. Standards as high as 1500 ng/mL in blood and 20,000 ng/g in mammary gland could be analyzed using lower injection volume or extrapolating the calibration curve beyond the upper limit of quantitation (mean %RE ≤±18.7; %RSD ≤2.2). Loss of AP occurred during overnight autosampler storage as well as frozen storage in as few as 15 days, but incorporation of IS prior to storage corrected for the loss such that calculated concentrations were within 84.7-117% of day 0 concentrations following frozen storage up to ≥32 days in both matrices. Matrix evaluation was performed in Hsd:Sprague Dawley®SD® rat and B6C3F1 mouse blood and mammary glands (mean %RE ≤±9.2; %RSD ≤4.3). These data demonstrate that the method is suitable for determination of AP in rodent blood and mammary glands.

Comparative analysis of platelet concentrates prepared after two hours and overnight storage of buffy coat at room temperature

IntroductionThe overnight storage of the buffy coat (BC) at room temperature has logistic and operational advantages for the blood centre. The present study aimed to evaluate the impact of an overnight hold (stored) of BC at room temperature in comparison with the 2-hour hold (fresh) of buffy coats on the platelet concentrate (PC) characteristics. MethodsA total of 60 BCs were included in the study, 30 PCs (fresh) were prepared after two hours holding time of the BCs and the other 30 PCs (stored) were prepared after the overnight BC storage at room temperature. The primary endpoint of PCs evaluation was the platelet yield, volume, pH, WBC count, RBC count, and platelet swirling in the PC and the secondary endpoints were glucose concentration, lactate, LDH, and sterility of the PCs. All the tests were performed on the day+1 of the blood collection. ResultsThere was no difference concerning the volume, RBC count, and swirling between the two groups (P>0.05). The PCs from the fresh BC had higher pH and glucose concentration (P<0.05). On the other hand, the overnight hold of BC produced higher platelet counts, WBC counts, lactate, and LDH levels (P<0.05). All the 60 PCs did not record any bacterial growth on the culture media for the sterility results. ConclusionThe overnight hold of BC produces a higher platelet yield with higher storage lesions. This may also allow better supervision, ensuring better quality control.

Contamination of TEM Holders Quantified and Mitigated With the Open-Hardware, High-Vacuum Bakeout System.

Hydrocarbon contamination plagues high-resolution and analytical electron microscopy by depositing carbonaceous layers onto surfaces during electron irradiation, which can render carefully prepared specimens useless. Increased specimen thickness degrades resolution with beam broadening alongside loss of contrast. The large inelastic cross-section of carbon hampers accurate atomic species detection. Oxygen and water molecules pose problems of lattice damage by chemically etching the specimen during imaging. These constraints on high-resolution and spectroscopic imaging demand clean, high-vacuum microscopes with dry pumps. Here, we present an open-hardware design of a high-vacuum manifold for transmission electron microscopy (TEM) holders to mitigate hydrocarbon and residual species exposure. We quantitatively show that TEM holders are inherently dirty and introduce a range of unwanted chemical species. Overnight storage in our manifold reduces contaminants by one to two orders of magnitude and promotes two to four times faster vacuum recovery. A built-in bakeout system further reduces contaminants partial pressure to below 10−10 hPa (Torr) (approximately four orders of magnitude down from ambient storage) and alleviates monolayer adsorption during a typical TEM experiment. We determine that bakeout of TEM holder with specimen held therein is the optimal cleaning method. Our high-vacuum manifold design is published with open-source blueprints, parts, and cost list.

Muscle water properties in raw intact broiler breast fillets with the woody breast condition

The purpose of this study was to investigate the relationships between muscle water properties, water-holding capacity (WHC), and woody breast (WB) severity in intact raw broiler breast fillets. Broiler pectoralis major deboned at 3 h postmortem was collected from a commercial plant and categorized as normal (NORM), moderate WB, or severe WB (SEV). Meat drip loss was calculated based on weight loss during overnight storage at 4°C. Water properties of the intact fillets were determined with time domain nuclear magnetic resonance and the T2 relaxation times were determined using an inverse Laplace algorithm (CONTIN). Three T2 water components, hydration water (T2b), intra-myofibrillar water (T21), and extra-myofibrillar water (T22), were identified. With increasing WB severity, the time constant of each water component and the relative content of T22 (P22) increased while the relative areas of T2b and T21 (P2b and P21, respectively) decreased. Spearman correlation analysis showed that there were significant correlations between the WB condition score and either the time constant or normalized area for each T2 component. T22 normalized areas (A22) were most strongly correlated with the WB score (r = 0.75); however, the weakest correlation was found between the WB score and T21 areas (A21). Pearson correlation analysis revealed that the strongest correlation (r = 0.64) was found between A22 and drip loss; however, there was no correlation between A21 and drip loss. Within the NORM group, drip loss was significantly correlated to the time constants for both T2b and T21. Within the SEV group, only A22 was significantly correlated to drip loss. These data indicate that the WB condition has a significant impact on the distribution of water within the intact muscle tissue. The content of extra-myofibrillar water in broiler breast fillets may be a key factor responsible for the poor WHC measurements in WB meat.

Folate Forms in Red Blood Cell Lysates and in Intact Washed Red Blood Cells Are Stable for a Few Days During Refrigerated Storage

Abstract Objectives We investigated whether folate forms are stable in washed red blood cells (RBC) stored refrigerated for up to 9 days relative to RBC-lysates in ascorbic acid under the same storage conditions. Methods We prepared washed RBCs from freshly collected EDTA blood (n = 6 donors). For RBC-lysate 1, we diluted RBCs with saline (1/2 dilution), mixed well and prepared lysates (1/11 dilution) with 1% ascorbic acid (2 vials/time point; n = 12 vials/donor) for storage at 4°C for ≤ 9 days (baseline, 1, 2, 3, 6, and 9). For RBC-lysate 2, we aliquoted ∼0.5 mL washed RBCs (1 vial/time point; n = 5 vials/donor) for storage at 4°C for ≤9 days (1, 2, 3, 6, and 9). When refrigerated storage of samples was completed, RBC lysate 1 samples were frozen at −70°C, while the washed RBCs were diluted with saline (1/2 dilution), mixed well and RBC-lysates 2 were prepared (1/11 dilution) with 1% ascorbic acid (2 vials/time point; n = 10 vials/donor). All samples were stored at −70°C until analysis. At the time of analysis, we processed samples (2 replicates/time point; n = 2 days) for folate polyglutamate deconjugation using recombinant exo γ-glutamyl hydrolase, and conducted sample clean-up by automated solid phase extraction prior to analysis by LC-MS/MS. Results We found negligible losses of major folate forms after overnight refrigerated storage, and folate losses gradually increased over time (∼5% by day 6). The loss of 5-methyltetrahydrofolate and total folate (mean ± SD) after 2 days of storage for RBC lysate 1 was 1.2% ± 1.5% and 1.0% ± 1.3%, and for RBC-lysate 2 was 2.2% ± 1.1% and 2.7% ± 1.9%, respectively. The baseline concentration of non-methyl folate (sum of minor folate forms: 5-formyltetrahydrofolate [&amp;lt;LOD], tetrahydrofolate, and 5, 10-methenyltetrahydrofolate) was small (&amp;lt;3.0 nmol/L) in both sample types, and we noticed a slight increase (∼10%) after overnight storage. 5,10-Methenyltetrahydrofolate appeared to gradually convert to tetrahydrofolate upon storage in RBC-lysate 2 samples. We observed no noticeable changes for MeFox in either sample type. Conclusions Overnight refrigerated storage of RBC-lysates or washed RBCs is feasible and avoids notable losses of folate forms. Prolonged refrigerated storage promotes interconversions of minor folate forms. Funding Sources This work was supported by direct appropriations from U.S. Congress.