Storage Temperature Research Articles

The Utility of an Human Papillomavirus Genotype Assay for Cancer Screening in Self-Collected Urine and Vaginal Samples from Japanese Women

Objectives: The high incidence of invasive cervical cancer among those who have not undergone cancer screening is a serious problem. This study aimed to investigate the utility of human papillomavirus (HPV) test results from self-collected urine and vaginal samples as screening tools. Design: The study was conducted in two steps. First, the appropriate storage container, temperature, and time until urine HPV assay performance were verified. Second, the results of spot urine testing under those conditions and of gynecologist-collected cervical and self-collected vaginal samples were compared to verify the feasibility of using the BD Onclarity® HPV assay for individuals with abnormal cervical cytology. Participants/Materials, Setting, Methods: The participants were 121 women with abnormal cervical cytology. Self-collected urine and vaginal samples, along with gynecologist-collected cervical samples, were tested for HPV using the BD Onclarity® HPV assay. The optimal conditions for urine sample storage were identified by comparing the HPV detection rates under various conditions. Results: Urine stored in a BD Probe Tec™ (QxUPT) for less than 72 h at room temperature was found to have the highest HPV positivity rate. Under these conditions, the detection rates of HPV in urine, cervical, and vaginal samples were examined. HPV type 16 was detected in 41.7% of the cervical samples, type 18 in 10%, and types 31 and 52 in 12.6% each. The concordance rate for HPV testing between clinician-collected cervical and urine samples was 63.9% (kappa: 0.34; 95% CI: 0.21–0.47), and that between clinician-collected cervical and self-collected vaginal samples was 77.8% (kappa: 0.68; 95% CI: 0.53–0.83), indicating good concordance. In a population with an HPV-related lesion/tumor prevalence of approximately 70%, the sensitivity of HPV testing was 82.7% for the cervix, 46.4% for urine, and 75.7% for vaginal samples. Limitations: The primary limitation is the lower detection rate of HPV in spot urine samples than in other sample types, indicating room for methodological improvement. The study’s findings are based on a specific population, which may limit generalizability. Conclusions: We investigated the optimal self-collected urine-to-testing time and temperature. Self-collected vaginal and urine HPV tests show moderate-high concordance with clinician-collected cervical HPV tests, suggesting their potential utility for women who do not undergo regular cancer screening. However, the sensitivity was not high in spot urine. Therefore, further large-scale studies are needed to verify these findings and optimize testing methods to encourage broader participation in cancer screening programs.

Comparative Study of Temperature and Pressure Variation Patterns in Hydrogen and Natural Gas Storage in Salt Cavern

Clarifying the distribution of temperature and pressure in the wellbore and cavern during hydrogen injection and extraction is crucial for quantitatively assessing cavern stability and wellbore integrity. This paper establishes an integrated flow and heat transfer model for the cavern and wellbore during hydrogen injection and withdrawal, analyzing the variations in temperature and pressure in both the wellbore and the cavern. The temperature and pressure parameters of hydrogen and natural gas within the chamber and wellbore were compared. The specific conclusions are as follows. (1) Under identical injection and withdrawal conditions, the temperature of hydrogen in the chamber was 10 °C higher than that of natural gas, and 16 °C higher in the wellbore. The pressure of hydrogen in the chamber was 2.9 MPa greater than that of natural gas, and 2.6 MPa higher in the wellbore. (2) A comparative analysis was conducted on the impact of surrounding rock’s horizontal and numerical distance on temperature during hydrogen and natural gas injection processes. As the distance from the cavity increases, from 5 to 15 m, the temperature fluctuation in the surrounding rock diminishes progressively, with the temperature effect in the hydrogen storage chamber extending to at least 10 m. (3) The influence of rock thermal conductivity parameters on temperature during the processes of hydrogen injection and natural gas extraction is also compared. The better the thermal conductivity, the deeper the thermal effects penetrate the rock layers, with the specific heat capacity having the most significant impact.

Standardization of Protocol for Production of Synthetic Seed of Brahmi (Bacopa monnieri) and Assessment of their Longevity under Differential Storage Condition

Brahmi [Bacopa monnieri (L) Pennel] is an important medicinal small succulent, medicinal herb belongs to the family ‘Scrophulariaceae’ which is commonly known as Jala Brahmi or Nira-Brahmi. National Medicinal Plants Board (NMPB), estimated an annual demand of Bacopa in 2004-2005 was 6621.8 tonnes, with a 7% annual growth rate. Micropropagation techniques and production of synthetic seeds can be a breakthrough to overcome the low availability of Brahmi propagating materials. So, the present study was conducted to standardize the protocol for preparation of synthetic seeds of Brahmi using nodal segment. The experiment was conducted with five treatments by using combinations of different concentrations of sodium alginate and calcium chloride and preparing two types of synthetic seeds (single layer and double layer). Further the longevity of both single layer and double layered synthetic seeds was studied under two different storage temperatures at 25°C (ambient condition) and 4°C (refrigerator) up to 40 days by inoculating them in two different types of media. The result showed no significant effect of concentrations of gelling agent and complexing agent on the germination percentage of synthetic seeds. The number of shoots per synthetic seed varied from 22.00 to 29.35 with 100% germination and maximum shoots/synthetic seed and shoot length was obtained by using T5 (single layered with MS media, 3% sodium alginate and 2.5% calcium chloride). Maximum number of days of storage can be observed in the single layer synthetic seeds when stored at 4°C along with media followed by double layer synthetic seeds when stored at 4°C along with media. However, seeds stored on vermiculite got dried up in the initial days of storage. Germination was found to be higher in seeds stored at 4°C when compared to seeds stored at 25°C. The study as a whole elucidated the opportunity for the use of synthetic seed as an alternative to traditional propagation system for Brahmi.

Multi-objective optimization and influence degree analysis of the thermally integrated HP-ORC carnot battery based on the orthogonal design method and grey relational analysis

Thermally integrated heat pump (HP)-organic Rankine cycle (ORC) Carnot battery is a promising solution for efficient energy storage and waste heat utilization. Firstly, in order to research the influence degree of the operating parameters on the system performance, single-objective optimization is conducted by the orthogonal design method. Then, multi-objective optimization is conducted by combining the grey relational analysis (GRA) and orthogonal design method to assess the comprehensive performance of the system. The importance order and contribution rates of the operating parameters and the optimal operating combinations are determined. Finally, in order to explore the influence mechanism of the parameters on the system performance, the influence of the parameters on the subsystems and the subsystems on the whole system is quantitatively analyzed. The results show that thermal storage temperature difference, waste heat temperature difference and cold source temperature are the most important parameters affecting the comprehensive performance of the whole system. Under the optimal operating conditions, round-trip efficiency, exergy efficiency, energy storage capacity (ESC), energy storage density (ESD) and levelized cost of storage (LCOS) are 47.05 %, 22.66 %, 1943 kW, 2.90 kWh/m3 and 0.38$/kWh, respectively. Compared to the ORC subsystem, the HP subsystem is more important to round-trip efficiency and exergy efficiency.

Impact of refrigeration of different Resin composite restorative materials on the marginal adaptation in class II restorations

BackgroundThe pre-polymerization temperature of resin composite restorative materials could influence their adaptation to cavity details. As a current debate is existing about the refrigeration of resin composite restorative materials, this study was designed to assess the effect of refrigeration of 3 types of resin composite restorative materials with different matrix systems on their marginal adaptation in Class II restorations.MethodsForty-two sound maxillary molars, each with two separated Class II cavities, were used in this study. The teeth were assigned into 3 main groups (n = 14) according to the restorative /adhesive system used; an Ormocer-based composite (Admira Fusion/Futurabond M+, Voco GmbH, Cuxhaven, Germany), a methacrylate modified Ormocer-based (Ceram.X SphereTEC One/Prime&Bond Universal, Dentsply Sirona GmbH Konstanz, Germany), and a methacrylate-based (Tetric N-Ceram/Tetric N-Bond Universal, Ivoclar Vivadent AG, Schaan, Liechtenstein). Each group was then divided into 2 subgroups (n = 14) according to the gingival margin location; 1 mm above and 1 mm below the cemento-enamel junction (C.E.J). Each subgroup was further divided into 2 categories (n = 7) according to the storage temperature; stored at room temperature or stored in refrigerator at 4°- 5° C. Epoxy resin replicas were observed under scanning electron microscope (SEM) to examine the marginal gaps. A gab scoring system was used to assess the marginal adaptation of each restoration by giving scores on the basis of measurements of the maximum marginal gaps. The data obtained were statistically analyzed using the Chi-square test at a significance level of p < 0.05.ResultsNone of the tested groups exhibited 100% gap-free margins irrespective of margin location or storage temperature. For both storage temperatures, no statistically significant difference was observed among all tested groups either with margins located above or below C.E.J (p > 0.05). As well, there was no statistically significant difference when comparing both marginal locations for each material (p > 0.05). Regarding the effect of storage temperature, statistically significant difference was only observed between the room-temperature stored groups with margins located above C.E.J and their corresponding groups stored in refrigerator (p < 0.05).ConclusionThe refrigeration of resin composite restorative materials prior to the restorative procedures revealed a deleterious effect on marginal adaptation of the restorations with margins located in enamel regardless the type of material used.

Microbiological quality of vegan alternatives to dairy and meat products in England during 2022-3.

Plant-based alternatives to meat and dairy products have become increasingly popular in the UK. Despite a public perception that they have a relatively low microbiological risk, outbreaks of illness have been linked with these foods. This study aimed to assess the microbiological safety and quality of vegan alternatives to dairy and meat products available in England. Samples were collected between September 2022 and March 2023 from retail, production, and catering premises, and tested for a range of bacterial pathogens and hygiene indicators using standard procedures. A total of 937 samples were tested, of which 92% were of a satisfactory microbiological quality, 3% were borderline, and 5% were unsatisfactory. Those interpreted as unsatisfactory were due to elevated counts of Enterobacteriaceae and Escherichia coli (indicators of poor hygiene) rather than pathogenic microorganisms. Listeria monocytogenes was present in five samples of tofu, all from the same producer (all at counts of<100 CFU g-1), while other Listeria species were detected at counts of<20 CFU g-1 in two burgers and two 'vegan chicken' products. The majority of samples did not have pH and water activity values that would significantly contribute to preventing microbial growth: 62.4% had pH>5.0 and 82.4% had Aw>0.94. The majority of vegan products examined were of a satisfactory quality, but results demonstrate that microbiological control must be maintained using appropriate processing and storage temperatures, and application of a safe length of shelf life.

Effect of concentration, temperature, and time on the stability of hepatitis C virus nucleic acid in serum.

Previously, there were few reports about the influence of different concentrations of sample nucleic acid on the stability of samples at various temperatures and times in various literatures. Therefore, it is necessary to analyze the influence of concentration factors on the stability of samples and test results at different storage times and temperatures. This study took the concentration of hepatitis C virus nucleic acid as the research object to further understand the stability of hepatitis C virus nucleic acid test samples under various storage conditions, to provide data reference for the treatment of hepatitis C virus nucleic acid and RNA test samples before clinical laboratory test, and provide guidance and help for the improvement of laboratory quality control.

Charging of an Air–Rock Bed Thermal Energy Storage under Natural and Forced Convection

An air-rock bed thermal storage system was designed for small-scale powered generation and analyzed with computational fluid dynamics (CFD) using ANSYS-Fluent simulation. An experimental system was constructed to compare and validate the simulation model results. The storage unit is a cylindrical steel container with granite rock pebbles as a storage medium. The CFD simulation used a porous flow model. Transient-state simulations were performed on a 2D axisymmetric model using a pressure-based solver. During charging, heat input that keeps the bottom temperature at 550 °C was applied to raise the storage temperature. Performance analysis was conducted under various porosities, considering natural and forced convection. The natural convection analysis showed insignificant convection contribution after 10 h of charging, as observed in both average air velocity and the temperature profile plots. The temperature distribution profiles at various positions for both convection modes showed good agreement between the simulation and experimental results. Additionally, both cases exhibited similar temperature growth trends, further validating the models. Forced convection reduced the charging time from 60 h to 5 h to store 70 MJ of energy at a porosity of 0.4, compared to natural convection, which stored only 50 MJ in the same time. This extended charging period was attributed to poor natural convective heat transfer, indicating that relying solely on natural convection for thermal energy storage under the given conditions is not practical. Using a small fan to enhance heat transfer, forced convection is a more practical method for charging the system, making it suitable for power generation applications.

Investigation of optimal sperm storage conditions for short-term storage

Objective The quality of sperm cells is important role in the success rates of assisted reproduction technology (ART) treatments. The quality of the sperm cells shows variations depending on the temperature of short-term semen storage as well as the methods of semen preparation. Thus, this study aimed to investigate the sperm viability, motility and DNA fragmentation following different sperm preparation methods and short-term storage conditions, respectively. Materials and Methods A total of 25 semen samples were evaluated. In the first part of this study, different incubation temperatures were investigated in two groups, in such the first group involved the semen samples and the second group involved the sperm cells separated by density gradient centrifugation method, respectively. The samples in each group were incubated at 4°C, room temperature (21°C) and 37°C for 24 hours, respectively. The sperm cell qualities were evaluated by mobility analysis, DNA fragmentation by acridine orange staining and sperm cell viability by propidium iodide staining. Results and Discussion The analysis outcome demonstrated that the mobility, DNA fragmentation and viability of the sperm cells were statistically different when incubated at RT (21°C) in both groups. Furthermore, samples prepared by the density gradient centrifugation method were shown to have better quality. The optimum short-term storage temperature was detected to be the room temperature. Conclusion The conclusion of this investigation is crucial to assess storage conditions in ART clinics. This study provides essential data for short-term sperm storage and preparation methods to improve the success rates of ART clinics. Bangladesh Journal of Medical Science Vol. 23 No. 04 October’24 Page : 1137-1141

The influence of storage conditions on hygiene condition and eicosapentaenoic acid oxidation in Nannochloropsis algae for sustainable food supply.

Nannochloropsis algae contain approximately 20% polyunsaturated fatty acids (PUFA) and hold significant potential for high-quality eicosapentaenoic acid (EPA) food industrialization. However, EPA in Nannochloropsis sp. is prone to oxidation, and microbial growth is a critical factor affecting the shelf life of fresh food. Storage composition and temperature are primary factors influencing microbial growth, yet these aspects are not fully understood. This study investigates the effects of temperature and encapsulation on EPA content in nano-products over time. Nano-powder and nanobeads derived from Nannochloropsis sp. served as raw materials. Additionally, changes in aerobic plate counts and coliform groups were monitored. The results indicated that nanobeads, due to their more complex processing and less mature packaging, were more susceptible to coliform contamination compared to nano-powder. In terms of EPA stability, nanobeads exhibited a longer storage life than nano-powder. The oxidation rate of both nano-powder and nanobeads was faster at 37 °C than at 25 °C. These findings can inform general shelf life estimation, rapid detection of total lipid content in nano-products and macro extraction of nano-oil. Moreover, they have significant implications for delaying EPA oxidation in nano-products and improving hygienic quality control for microbial detection. © 2024 Society of Chemical Industry.

A-087 Stability of fecal calprotectin extracted with BÜHLMANN CALEX® Cap

Abstract Background Calprotectin, a granulocyte-derived alarmin protein, is an established noninvasive biomarker of intestinal mucosal inflammation. Fecal calprotectin testing has become the gold standard for diagnosis of inflammatory bowel diseases. The aim of this work was to determine the stability of calprotectin in stool extracts prepared with the BÜHLMANN CALEX® Cap stool preparation device. Methods The analyte stability study was carried out using CALEX® Cap extracts of human fecal left-over samples. Extraction was performed for 24 hours at room temperature and samples were aliquoted and frozen at &amp;lt; -20°C. The baseline calprotectin concentration was measured from thawed aliquots, which were moved to target storage temperatures (2-8 °C, 18 °C, 28 °C, and 37 °C) for up to 22 days. Five stool samples covering the measuring range of the assay were analyzed with the BÜHLMANN fCAL® turbo in four replicates. Stability evaluations were performed according to CLSI guideline EP25-A1 and EP25Ed2. Results CALEX® Cap derived extracts stored refrigerated at 2-8°C for up to 20 days and at room temperature (18-28°C) for up to 6 days demonstrated deviations from baseline of less than 20%. CALEX® Cap derived extracts stored up to 15 days at 2-8°C and up to 2 days at room temperature (18-28°C) demonstrated deviations from baseline of less than 10%. Conclusions CALEX® Cap provides a stool collection method that stabilizes fecal calprotectin concentrations for an increased processing window. This allows significant timely separation between stool collection and the actual measurement of calprotectin. This specimen stabilization provides for accurate measurements for days after sample collection and thus allows for flexibility for diagnostic laboratories.

Influence of Hematocrit and Storage Temperature on the Stability of Dried Blood Samples in Serological Analyses of Tetanus, Diphtheria, and Pertussis.

Dried blood spots (DBSs) enable the study of serological markers of various pathogens without the need to obtain serum/plasma through venipuncture. Sixty-four blood samples were prepared on Whatman™ 903 cards using specimens obtained by venipuncture to study the detection of serological markers of diphtheria, tetanus, and pertussis in DBSs, and their stability 4 months post-collection. An automated chemiluminescent immunoassay was used to investigate diphtheria, tetanus, and pertussis IgG levels from both DBSs and plasma samples. An optimal cut-off value for DBSs was calculated to improve the performance of diphtheria and tetanus serological markers in DBSs, achieving high sensitivity (95% and 98%, respectively) and specificity (91.7% and 92.3%, respectively). No protection against pertussis was found in the population studied. The correlation observed between the plasma and the DBSs processed after sample collection was high (0.967-0.970) for all antibodies studied except pertussis (0.753), both considering hematocrit before sample elution or not. The correlation between DBSs and plasma for diphtheria and tetanus remained strong following a 4-month delay in DBS processing at 4 °C (0.925-0.964) and -20 °C (0.924-0.966), with only a minor decrease observed for diphtheria at room temperature (0.889), while maintaining a strong correlation for tetanus (0.960). For pertussis, the correlation between DBSs and plasma was drastically reduced after delaying its processing for 4 months at any temperature. To summarize, hematocrit levels within the normal range do not affect the processing of DBSs in the study of serological markers of diphtheria, tetanus, and pertussis. The DBS stability for serological diagnosis of diphtheria and tetanus is adequate when samples are stored at -20 °C for a period of 4 months. The pertussis serological marker does not appear to remain stable after 4 months, even when the DBS is stored frozen at -20 °C.

Microbiological profile and prevalence of histamine-producing bacteria in fresh sardines stored at different temperatures

Background and Aim: The European pilchard (Sardina pilchardus) is an important fish species for the Moroccan economy in terms of production and export. Biogenic amine histamine is a metabolite produced in the flesh of some fish species after death due to the decarboxylation of free histidine by histaminogenic bacteria. Failure to control the histamine risk in European pilchard may lead to public health and socioeconomic issues. This study aimed to estimate the prevalence of histaminogenic bacteria in association with histamine levels and the growth of microflora in Moroccan sardines (European pilchard). Materials and Methods: We conducted the study by monitoring Moroccan sardines of histamine content and microbiological profile (aerobic plate count [APC], coliforms, and thermo-tolerant coliforms [TTC]) during 6 days of storage at three different temperatures (0°C, 10°C, and ambient temperature [22°C]). The histamine assay was performed using a spectrofluorometric method, and the microbiological identification of histamine-producing bacteria was performed using a combination of biochemical and molecular tests. Results: The histamine content in European pilchard stored at 0°C was negligible. However, high concentrations were observed at 10°C and 22°C. The microbiological profile showed a positive association between microflora counts and histamine content according to storage time. At 0°C, a moderate increase in the APC, a decrease in coliforms, and an absence of TTC were observed. The rapid proliferation of all microflora was observed at 10°C, whereas at 22°C, the proliferation was almost exponential. Bacterial identification revealed the exclusive presence of species belonging to the Enterobacteriaceae family at varying frequencies depending on storage temperature. Morganella morganii and Proteus mirabilis had the highest histamine induction rates in L-histidine-supplemented broth, with 1600 and 255 parts per million (ppm), respectively, after 48-h incubation at 35°C. Klebsiella ozaenae could produce 136 ppm and Serratia plymuthica 115 ppm. Reverse transcription polymerase chain reaction showed positive results for the presence of genes associated with histidine decarboxylase. The hdc genes of M. morganii, P. mirabilis, and K. ozaenae were successfully amplified and exhibited strong similarity with the reference gene of M. morganii. Conclusion: This study describes for the first time the hdc gene in bacteria that form histamine in Moroccan sardines. The results also confirm that respect for the cold chain integrity is a crucial factor in histamine management. This information should help stakeholders in the implementation of sound strategies for managing the hazards associated with seafood and their products. Keywords: Enterobacteriaceae, histamine, Histaminogenic bacteria, polymerase chain reaction, sardine, seafood.

Shedding Light on Insulation: Probing the Varied Effects of Illumination and Storage Temperatures on Vegetable Oil Quality

Shedding Light on Insulation: Probing the Varied Effects of Illumination and Storage Temperatures on Vegetable Oil Quality

Environmental Outcomes of Reducing Medication Waste by Redispensing Unused Oral Anticancer Drugs

Medications are associated with substantial environmental outcomes, yet frequently end up being unused by patients. Waste-minimizing interventions, such as redispensing of quality-approved oral anticancer drugs remaining unused by patients at home, could reduce the environmental footprint of cancer treatment. To assess the environmental outcomes of redispensing quality-assured oral anticancer drugs and to explore how redispensing could be environmentally optimized. In this quality improvement study, a cradle-to-grave life cycle assessment was performed in the outpatient pharmacy of 4 Dutch hospitals, based on a prospective multicenter trial comprising 1071 patients with a clinical diagnosis of cancer and an active prescription for an oral anticancer drug stored at room temperature from February 1, 2021, to February 1, 2023, with a follow-up of 12 months per patient. Participants received prescribed oral anticancer drugs with additional quality-assurance materials (ie, seal bags and time-temperature indicators), so the pharmacy could redispense quality-assured drugs based on authenticity, appearance, remaining shelf life, and/or adequate storage. The estimated environmental outcomes avoided due to waste reduction (ie, production and transport and incineration of redispensed oral anticancer drugs) corrected for outcomes of process burdens (ie, quality assurance materials), quantified in 3 outcome measures: human health damage (disability-adjusted life-years), ecosystems damage (species × year), and climate change (kg of carbon dioxide equivalent [CO2-eq]) per patient per year. A volunteer sample of 1071 patients (median age, 70 years [IQR, 62-75 years]; 622 men [58.1%]) participated in the intervention. Redispensing oral anticancer drugs was initially associated with an environmental burden, mainly because of the high impact of time-temperature indicators. However, when quality-assurance materials were selectively used for temperature-sensitive oral anticancer drugs (ie, maximum storage temperature of 25 °C), redispensing was environmentally beneficial to human health and ecosystems, providing estimated climate benefits of 1.9 kg (95% CI, 1.4-2.6 kg) of CO2-eq per patient per year. In this quality improvement study, redispensing unused oral anticancer drugs was found to be a suitable strategy to reduce waste and improve environmental sustainability of cancer treatment after process optimization. Redispensing unused oral anticancer drugs could contribute to sustainability of cancer treatment through reduced costs and environmental outcomes.

Evaluation of extraction and storage conditions for quantification and characterization of silver nanoparticles in complex samples by single particle-ICP-MS

The extraction of nanoparticles (NPs) from complex matrices and subsequent storage can potentially alter the NPs physicochemical properties and hinder cross-study comparisons. Most NP extraction methods are designed and tested at high nanoparticle concentrations, although (eco)toxicological and regulatory monitoring programs require methods capable of analyzing NPs at environmentally relevant concentrations (lower ppb range). In this study, we investigated how extraction methods affect the characteristics of PVP coated and citrate-stabilized silver NPs (AgNPs) spiked into soil, sewage sludge, and biological samples at environmentally relevant concentrations using Single Particle Inductively Coupled Plasma Mass Spectrometry spICP-MS). Further we investigated the impact of storage temperature (-80°C to 21°C) and storage duration (1-28 days) on the particle characteristics such as particle size.We found that aqueous AgNPs samples with low ionic strength media retained their original characteristics (like particle size, particle concentration and particle-based Ag mass) when preserved at 4°C for up to 28 days. AgNPs dispersed in high ionic strength media were however better preserved at -80°C. Among the extraction agents, tetrasodium pyrophosphate was more efficient in extracting AgNPs from soil and sewage sludge matrices, while Proteinase K was more suitable for biological samples from organisms (earthworms or fish).Although our study focused only on AgNPs, it provides crucial information to aid interlaboratory comparisons and data interpretation for (eco)toxicological studies.

Optimal preservation condition for the extraction of cell-free DNA from bile of patients with biliary tract cancer.

Patients with biliary tract cancer (BTC) often have dismal outcomes due to the poor performance of traditional methods for early diagnosis. Recently, bile cell-free DNA (cfDNA) has been reported as a potential liquid biopsy material for BTC diagnosis. However, bile is a complex alkaline aqueous medium, and the proper storage conditions for bile remain to be explored. The aim of this study is to explore the effects of storing bile under various conditions on the stability of bile cfDNA and to determine the optimal conditions, thereby establishing a foundation for the subsequent application of bile cfDNA in liquid biopsy for early diagnostic and prognosis monitoring of patients with malignant BTC. We evaluated the storage temperature and storage time for the preservation of bile samples. Bile samples were collected in cfDNA tubes with protectant covered inside or regular tubes without, and the stability of bile cfDNA was analyzed during 10 days at room temperature (RT) or after 2 months of storage at low temperatures. Bile cfDNA remained stable for bile samples being collected with cfDNA tubes and stored for 10 days at RT, while degraded with time for the case with regular tubes. When bile samples were collected with cfDNA tubes and stored for 2 months at 4 ℃, bile cfDNA remained stable, however, if collected with regular tubes, bile cfDNA exhibited a slight loss of integrity. No significant difference was observed for 2 months storage at -20 or -80 ℃. Our findings suggested that for bile cfDNA research, bile samples should be collected with cfDNA tubes and it can be transported for short-term shipment at RT, and could be stored at 4 ℃ with cfDNA tubes, or frozen at -20 ℃ with regular tubes.

Impact of formulation and home storage conditions on the content of furan and derivatives in powdered infant formula

While autoxidation of Polyunsaturated Fatty Acids (PUFAs) is a potential source of furan and its derivatives, the regulatory obligation to enrich powdered infant formulae (PIF) with some of these compounds might raise safety issues. The aim of this study was to investigate the impact of formulation and home storage conditions on the generation of furan and its derivatives in PIF. Furan, 2-methylfuran (2-MF) and 3-methylfuran (3-MF) were monitored by a validated SHS-GC-Q Exactive-Orbitrap MS method in six PIF formulated with high or low concentrations of different PUFAs (ALA, ARA, DHA), pro-oxidants (iron) and anti-oxidants (vitamins) and stored for 21 days under more or less oxidizing home storage conditions, including temperature (19 °C or 40 °C) and oxygen exposure (protected or not from ambient air). For the three compounds, the quality of the measurements in PIF was first confirmed with suitable linearity, limits of quantification, precision and recoveries. In a second step, the impact of the formulation was evaluated on six PIF commercial samples differing significantly in their composition in ALA, ARA, DHA, iron and/or vitamin E and C. While furan was quantifiable (2.8 µg/kg) in only one PIF with high content of DHA and iron and low level of vitamins, 2-MF was only quantifiable in two other PIF which were distinguished by a 15 to 20 % higher ALA content and 3-MF was not detected in any of the six PIF studied.. In a third step, the effect of storage conditions was studied on the six PIFs and the increase in storage temperature to 40 °C led to an increase of up to 33 % in the 2-MF concentration in the two PIF formulations where it was quantifiable. Nevertheless, the estimation of the Margin of Exposure (MOE) showed that the risk related to furan and its derivatives could be ruled out regardless of the formulation or storage conditions. Finally, in order to explore other derivatives and to investigate the mechanisms involved in the generation of furan compounds in PIF, a suspect screening approach was implemented on the SHS-GC-Q Exactive Orbitrap MS data. It made it possible on the one hand to point out two additional furan derivatives in PIF, 2-ethylfuran and 2-pentylfuran. On the other hand, it enabled to discuss the oxidation pathways involved in the generation of furan compounds in PIF from profiling of known PUFA oxidation markers also detected in the samples.

Evaluation of Postharvest Quality of Fresh Kesum Leaves (Persicaria minor) during Storage in Different Temperatures and Packaging

Persicaria minor, locally known as kesum, is an herb synonym in Southeast Asia as a food flavor enhancer and seasoning for Eastern delicacies. Kesum leaves are quickly wilted, shrink, and lose moisture once harvested. This study aimed to determine optimum parameters: storage temperature and suitable packaging to maintain the freshness and quality of kesum and prolong the storage life. This study consisted of two activities: determining the best storage temperature and determining suitable packaging. For the first activity, kesum leaves were harvested manually from matured plants, transported to a packinghouse, sorted, cleaned, packed, and sealed in polyethylene (PE) 0.04mm before being stored at different temperatures, which were 5°C, 8°C, 10°C and 13°C, while for the second activity, kesum leaves were packed and sealed in different packaging: polyethylene (PE) 0.04mm and polypropylene (PP) 0.04mm and stored at 5°C. The postharvest quality of all samples and treatments (weight, moisture content, pH, total titratable acidity, total soluble solids, respiration rate of the leaves, total phenolic content, and leaf color values) was measured and recorded weekly throughout the storage period. The study's results found that storing fresh kesum leaves at 5°C temperature and in PE 0.04mm packaging can delay the senescence and maintain the quality and postharvest attributes for up to 35 days, as shown in lower respiration rate, maintain the pH, total titratable acidity, total soluble solids and color of the leaves, and show higher total phenolic content for up to 35 days of storage compared to storing in temperature of 8°C, 10°C, and 13°C and PP 0.04mm packaging.

Surface sterilization and moist storage conditions for recalcitrant seeds of three Asian tree species

Ex situ storage of recalcitrant seeds is challenging, due to seed-desiccation sensitivity. Various moist storage techniques have been used to store seeds, but moist conditions also promote fungal infection. This study determined suitable sterilizing agents for surface cleaning and suitable moist storage conditions for three tree species: Heynea trijuga Roxb. ex Sims, Neocinnamomum caudatum (Nees) Merr., and Phoebe lanceolata (Nees) Nees. We used a factorial design with three factors: sterilizing agents, storage temperature and storage media. There were four surface-sterilization treatments: no sterilization (control), 70% ethanol, 3% NaOCl, and 25% metalaxyl. Seeds were stored with two storage media (moist sand and moist filter paper) and at two storage temperatures (room temperature and 4 °C). Percentage fungal infection and seed germination were recorded. For H. trijuga and P. lanceolata, 3% NaOCl (five-minute soaking) was the best sterilizing agent, whilst N. caudatum had no optimal sterilizing agent. We recommend storing seeds with moist storage media at 4 °C. The best storage media for H. trijuga was moist filter paper, while for P. lanceolata either moist sand or filter paper was suitable. However, N. caudatum seeds should be sown immediately after collection. Our findings emphasize the importance of testing individual species to determine optimal techniques for surface sterilization and storage.