Alternative Preservation Method Research Articles

Application of Quantitative Magnetic Resonance Imaging (QMRI) to Evaluate the Effectiveness of Ultrasonic Atomization of Water in Truffle Preservation.

Truffles of the Tuber genus (Pezizales, Ascomycetes) are among the most valuable and expensive foods, but their shelf life is limited to 7-10 days when stored at 4 °C. Alternative preservation methods have been proposed to extend their shelf life, though they may alter certain quality parameters. Recently, a hypogeal display case equipped with an ultrasonic humidity system (HDC) was developed, extending the shelf life to 2-3 weeks, depending on the truffle species. This study assesses the efficacy of HDC in preserving Tuber melanosporum and Tuber borchii ascomata over 16 days, using quantitative magnetic resonance imaging (QMRI) to monitor water content and other parameters. Sixteen T. melanosporum and six T. borchii ascomata were stored at 4 °C in an HDC or a static fridge (SF) as controls. QMRI confirmed that T. borchii has a shorter shelf life than T. melanosporum under all conditions. HDC reduced the rate of shrinkage, water, and mass loss in both species. Additionally, the Apparent Diffusion Coefficient (ADC), longitudinal relaxation time (T1), and transverse relaxation time (T2), which reflect molecular changes, decreased more slowly in HDC than SF. QMRI proves useful for studying water-rich samples and assessing truffle preservation technologies. Further optimization of this method for industrial use is needed.

Evaluation of freezing, ultra-cold freezing, and freeze-drying on the main components of human breast milk

Breastfeeding is the ideal food during the first two years, but only 31 % of children under six months of age receive exclusive breastfeeding, below the global recommendation. Alternative methods to direct breastfeeding have been used, including the creation of personal milk banks and the practice of deferred breastfeeding. This research aimed to evaluate the changes in human breast milk composition under freezing, ultra-cold freezing, and freeze-drying conditions in terms of: 1) carbohydrate profile as lactose and Human Milk Oligosaccharides; 2) protein quantity and characteristics (total crude protein, free amino nitrogen, in vitro digestibility, electrophoretic profile and secondary structure), and 3) functional and physicochemical components/ characteristics as pH, density and moisture, as well as pro- and anti-inflammatory cytokines and levels of secretory immunoglobulin A. With different preservation methods, no differences were detected in pH, density, moisture, total crude proteins, cytokines, four Oligosaccharides (3’-Sialyllactose, 6’-Sialyllactose, Lacto-N-Tetraose, and Lacto-N-neoTetraose), and secretory immunoglobulin A. The immunological components remain stable when frozen at –20 and −80°C and during freeze-drying, allowing women to prolong breastfeeding without changing the carbohydrate, protein, and functional or physicochemical main profile. Our results improve the understanding of human breast milk quality when alternative preservation methods are required beyond direct breastfeeding, aiming to extend the duration and rates of breastfeeding.

Tape Stripping Method in Dermatological and Pharmacological Research: Evaluating Fresh and Frozen-Thawed Porcine Skin.

Background The stratum corneum (SC) plays a crucial role in protecting the skin and regulating water loss. Tape stripping is a well-established method for studying skin barrier function and evaluating topical treatments. However, the behavior of fresh versus frozen-thawed skin during tape stripping has not been extensively compared. Objective This study aims to compare the removal of the stratum corneum from fresh and frozen-thawed porcine skin using tape stripping. It also aims to assess the reliability of tape weighing versus histological methods in quantifying SC removal. Methods Fresh and frozen-thawed porcine ears were obtained, cleaned, and subjected to tape stripping at varying numbers of strips from zero to 40. Tape weight and histological measurements were used to quantify SC removal. Statistical analyses were conducted to compare SC thickness and tape weight between the two types of skin. Results The study found that frozen-thawed skin exhibited a non-linear rate(r = 0.65) of SC removal per tape strip in the first five strips compared to a linear removal for fresh skin (r = 0.96). By the fifth tape strip, frozen-thawed samples had lost 80.6% of their SC, while fresh samples had only lost 33.5% (P < 0.03). Tape weighing and histological measurements showed strong correlations (r = 0.93 for fresh skin and r = 0.95 for frozen-thawed skin), indicating that tape weighing is a reliable alternative to histology for assessing SC removal on both sample types. Conclusions Fresh and frozen-thawed porcine skin respond differently to tape stripping, with frozen-thawed skin showing accelerated SC removal in the first five strips. The strong correlation between tape weighing and histological analysis supports the use of tape weighing as a practical tool for evaluating SC removal. These findings have implications for specimen selection and methodological standardization in dermatological and pharmacological research. Future research should explore alternative preservation and SC thickness measurement methods and their impact on tape stripping outcomes.

Extending Cucumber Shelf Life: A Comparative Study of Low-Cost Drip Cooling Systems Using Different Fabric Coverings

Aims: Postharvest losses of cucumbers pose significant challenges for small-scale farmers in developing regions due to limited access to modern cold storage. This study investigated the effectiveness of low-cost indigenous drip coolers with different fabric coverings as an alternative preservation method. Study Design: Completely Randomized Design (CRD) with five treatments and three replications. Place and Duration of Study: Department of Crop Science, Surigao del Norte State University – Mainit Campus, Mainit, Surigao del Norte, Philippines on March 1-20, 2024. Methodology: Cucumber fruits were stored in indigenous drip coolers constructed with various recycled fabric coverings. Treatment combinations were: T1 - Control ambient temperature, T2 - Alpha Gina fabric, T3 - Silk satin bedsheets, T4 – Cotton comforter, and T5 - Cotton Blanket. The study evaluated color change, firmness, shriveling index, visual quality rating, weight loss, and damage rate over 20 days storage period. Results: Significant differences in cucumber fruit quality emerged at different time points across various parameters. Differences in color change and firmness were observed on day 15 (p = 0.023), while shriveling and visual quality showed differences by day 20 of storage (p = 0.042 and p = 0.042, respectively). However, in terms of weight loss, the Alpha gina fabric significantly had the highest loss among fabric covers used (p = 0.038). Consistently, the control ambient group exhibited the poorest quality, while treatments like the cotton comforter and silk satin bedsheet were most effective in preserving cucumber quality. Conclusion: This study demonstrates the effectiveness of indigenous drip coolers in preserving cucumber quality, with silk satin bedsheets and cotton comforter coverings consistently outperforming other materials and ambient conditions. These coolers were effective in slowing color change, maintaining firmness, reducing shriveling, and preserving overall visual quality. The study highlights the importance of fabric selection in drip cooler design, with cotton comforter and silk satin bedsheet coverings likely offering better air circulation, moisture retention, and insulation.

A Comprehensive Review of Donor Corneal Preservation Strategies.

Successful corneal transplantation relies on the viability of the corneal endothelium. Although various preservation systems have been developed in the field of eye banking, long-term storage of the corneal endothelium poses challenges and is costly. Optisol-GS in the past has been the most commonly used solution for intermediate-term corneal storage in the United States. However, disruptions in the availability of Optisol-GS, caused by rising costs and supply shortages, have necessitated alternative methods of corneal preservation. Previously described preservation methods include hypothermia (2-8°C) for short-term storage (7-14 days), organ culture (28-37°C) for intermediate storage (4-7 weeks), and cryopreservation for longer-term storage. In this review, we examine standard practice and alternative methods for corneal storage.

Viability and virulence of freeze-dried Beauveria bassiana, Metarhizium acridum, and Hirsutella thompsonii isolates under non-refrigerated conditions in Ghana

ABSTRACT The necessity of cold storage for mycoinsecticides poses a logistical challenge in tropical Africa, where maintaining consistent refrigeration is a major obstacle. To investigate alternative preservation methods, our study evaluates conidial-based freeze-dried formulations of Beauveria bassiana (IMI 389521), Metarhizium acridum (IMI 330189), and mycelial-based freeze-dried formulations of Hirsutella thompsonii (IMI 327488 and IMI 391722). Propagules were cryoprotected with lactose, mannitol, and raffinose, encapsulated, and freeze-dried in plastic and glass vials. Bioformulations in glass vials were cryoprotected with mannitol only. Viability (CFU/g each day) of bioformulations, following 16-weeks-storage, and their virulence (at 1 × 107 conidia mL−1 in six days) against key pests of maize and cassava in Africa (larger grain borer, variegated grasshopper and cassava green mite) were determined under ambient laboratory conditions in Ghana. Our results show that non-freeze-dried conidia of B. bassiana and M. acridum had significantly higher viability and efficacy than freeze-dried versions, when stored under non-refrigerated conditions (averaging 27.7°C, 78.8% RH). Freeze-dried H. thompsonii mycelia were neither viable nor virulent against cassava green mite. Freeze-dried B. bassiana and M. acridum in plastic vials remained viable for 12 weeks, while in glass vials they were viable for seven weeks. Freeze-dried B. bassiana in both plastic and glass vials caused 38–43% mortality in larger grain borer, and M. acridum caused 45–82% mortality in variegated grasshopper. Further, conidia cryoprotected with lactose had significantly higher viability and mortality than raffinose and mannitol. Our research contributes to mycoinsecticide development, particularly in the context of storage and application challenges in tropical climates.

Effects of static magnetic field-prolonged supercooling preservation on blueberry quality

Although refrigeration and freezing are commonly used methods for preserving blueberries, refrigeration is only suitable for short-term preservation (storage at 4 °C for approx. 1 month), whereas ice crystals that form during freezing can lead to a degradation of blueberry quality. However, supercooling, which combines the advantages of freeze-free low-temperature storage, offers a novel alternative preservation method. In this study, we demonstrate that 10 mT static magnetic field treatment reduces the nucleation temperature of blueberries to below −6 °C and enhances their supercooling. The combination of static magnetic and temperature fields was found to prolong the effective storage of supercooled blueberries for up to 35 days, during which time, we detected no ice nucleation. Compared with refrigeration and freezing, the firmness of the supercooled-preserved blueberries increased by 24.1% and 31.2%, respectively, with corresponding reductions of 3.8% and 4.69% in weight loss. These changes were associated with a retardation of the decline in organic acid and soluble solid contents, along with the maintenance of high total phenolic and anthocyanin contents, antioxidant capacity, and the activities of polyphenol peroxidases and peroxidases. Collectively, our findings in this study indicate that supercooling would be an effective new approach for the preservation of blueberries.

Ex-vivo microscopic oncotesticular sperm extraction: step-by-step surgical technique at time of radical orchiectomy

To demonstrate the intraoperative surgical techniques required for simultaneous radical orchiectomy and microscopic oncotesticular sperm extraction (m-OncoTESE) in a step-by-step fashion. Video presentation. University Hospital (University of Chicago). A 37-year-old man (status after right orchiectomy at another institution for stage II-C testicular seminoma with positive preoperative tumor markers) was referred for contralateral orchiectomy of multifocal left testis mass and fertility preservation. Semen analysis before, microscopic testicular sperm extraction during, and semen or testicular specimen analysis after the first orchiectomy were unable to identify any sperm. A postoperative analysis of the m-OncoTESE performed on the left testis resulted in the cryopreservation of 200,000 motile sperm for future assisted reproductive technology (i.e., invitro fertilization or invitro fertilization-intracytoplasmic sperm injection). Left radical orchiectomy and left m-OncoTESE. A comprehensive visual documentation of m-OncoTESE surgical techniques with concurrent commentary detailing the reasons behind each surgical step. A brief discussion on the background of m-OncoTESE and alternative fertility preservation methods accompanies the procedure. This video provides a step-by-step guide to performing an m-OncoTESE (proceeding a radical orchiectomy in a patient with testicular cancer) as a means of fertility preservation in an azoospermic patient. Successful extraction and cryopreservation of testicular spermatozoa were achieved after targeted ex-vivo testicular microdissection. Sperm extraction via m-OncoTESE is a viable option for azoospermic patients with testicular cancer undergoing radical orchiectomies. The use of preoperative imaging and microsurgical techniques facilitates and optimizes surgical dissection and sperm recovery.

Breast milk preservation: thermal and non-thermal processes and their effect on microorganism inactivation and the content of bioactive and nutritional compounds.

Human Breast Milk (HBM) is widely acknowledged as the best nutritional source for neonates. Data indicates that, in 2019, 83.2% of infants in the United States received breast milk at birth, slightly reducing to 78.6% at 1 month. Despite these encouraging early figures, exclusive breastfeeding rates sharply declined, dropping to 24.9% by 6 months. This decline is particularly pronounced when direct breastfeeding is challenging, such as in Neonatal Intensive Care Units (NICU) and for working mothers. Given this, it is vital to explore alternative breast milk preservation methods. Technologies like Holder Pasteurization (HoP), High-Temperature Short-Time Pasteurization (HTST), High-Pressure Processing (HPP), UV radiation (UV), and Electric Pulses (PEF) have been introduced to conserve HBM. This review aims to enhance the understanding of preservation techniques for HBM, supporting the practice of extended exclusive breastfeeding. It explicitly addresses microbial concerns, focusing on critical pathogens like Staphylococcus aureus, Enterococcus, Escherichia coli, Listeria monocytogenes, and Cytomegalovirus, and explores how various preservation methods can mitigate these risks. Additionally, the review highlights the importance of retaining the functional elements of HBM, particularly its immunological components such as antibodies and enzymes like lysozyme and Bile Salt Stimulated Lipase (BSSL). The goal is to provide a comprehensive overview of the current state of HBM treatment, critically assess existing practices, identify areas needing improvement, and advocate for extended exclusive breastfeeding due to its vital role in ensuring optimal nutrition and overall health in infants.

Two-day Static Cold Preservation of α1,3-Galactosyltransferase Knockout Kidney Grafts Before Simulated Xenotransplantation.

Transplantation remains the preferred treatment for end-stage kidney disease but is critically limited by the number of available organs. Xenografts from genetically modified pigs have become a promising solution to the loss of life while waiting for transplantation. However, the current clinical model for xenotransplantation will require off-site procurement, leading to a period of ischemia during transportation. As of today, there is limited understanding regarding the preservation of these organs, including the duration of viability, and the associated molecular changes. Thus, our aim was to evaluate the effects of static cold storage (SCS) on α1,3-galactosyltransferase knockout (GGTA1 KO) kidney. After SCS, viability was further assessed using acellular sub-normothermic ex vivo perfusion and simulated transplantation with human blood. Compared to baseline, tubular and glomerular interstitium was preserved after 2 days of SCS in both WT and GGTA1 KO kidneys. Bulk RNA-sequencing demonstrated that only eight genes were differentially expressed after SCS in GGTA1 KO kidneys. During sub-normothermic perfusion, kidney function, reflected by oxygen consumption, urine output, and lactate production was adequate in GGTA1 KO grafts. During a simulated transplant with human blood, macroscopic and histological assessment revealed minimal kidney injury. However, GGTA1 KO kidneys exhibited higher arterial resistance, increased lactate production, and reduced oxygen consumption during the simulated transplant. In summary, our study suggests that SCS is feasible for the preservation of porcine GGTA1 KO kidneys. However, alternative preservation methods should be evaluated for extended preservation of porcine grafts.

Drying as an effective method to store soil samples for DNA-based microbial community analyses: a comparative study

Soil sampling for environmental DNA in remote and semi-remote locations is often limited due to logistical constraints surrounding sample preservation, including no or limited access to a freezer. Freezing at − 20 °C is a common DNA preservation strategy, however, other methods such as desiccation, ethanol or commercial preservatives are available as potential alternative DNA preservation methods for room temperature storage. In this study, we assessed five preservation methods (CD1 solution, 95% Ethanol, Dry & Dry silica gel packs, RNAlater, LifeGuard) along with freezing at − 20 °C, against immediate extraction on organic and mineral soils for up to three weeks of preservation. We assessed direct effects on DNA concentration and quality, and used DNA metabarcoding to assess effects on bacterial and fungal communities. Drying with Dry & Dry led to no significant differences from immediate extraction. RNAlater led to lower DNA concentrations, but effects on community structures were comparable to freezing. CD1, LifeGuard and Ethanol either caused immediate significant shifts in community structure, degradation of DNA quality or changes in diversity metrics. Overall, our study supports the use of drying with silica gel packs as a cost-effective, and easily applied method for the short-term storage at room temperature for DNA-based microbial community analyses.

The role of oxygenation in kidney and liver machine perfusion

Background. Organ transplantation is the optimal decision for patients in the end stage of many diseases. Certain conditions are required for the transportation and preservation of a donor organ after explantation, including factors such as temperature, pressure, and preservative solution. All currently available methods of preservation of donor organs are aimed at maximizing the complete preservation of the functional state of the graft from the moment of its removal to implantation and reperfusion in the recipient's body.Aim. The purpose of this review is to provide up-to-date information on the results of the studies performed in order to decide on the preferred method of organ preservation.Material and methods. An analysis of literature sources in English and Russian from 2009 to 2023 on this topic was performed in the databases PubMed, MEDLINE, Google Scholar. The review highlights the results of preclinical (on animal models) and clinical studies, as well as achievements in the field of ex-vivo machine perfusion with an emphasis on machine hypothermic perfusion and modified oxygenated hypothermic machine perfusion, subnormothermic machine perfusion and machine normothermic perfusion.Results. The daily increase in the number of patients in need of organ transplantation delays the timely selection and search for a donor. Organ donation after cardiac death is a promising step in an attempt to overcome the disbalance between the number of patients and organs, but the risk of developing early graft damage increases. The criteria for selecting donors and donor organs are being expanded, as a result, elderly donors and not-optimal grafts are included, but they are less resistant to ischemic damage. In this connection, there is a need for long-term infusion support through machine perfusion.Conclusion. In recent years, research has focused on alternative preservation methods, studying hypothermic, subnormothermic and normothermic machine perfusion. The use of machine perfusion has become the most widespread among kidney transplants and has shown good results. Further development is expected in the field of studying and improving this method of organ preservation, which allows not only transporting, but also improving the functional state of the graft.

Primordial Germ Cell Cryopreservation and Revival of Drosophila Strains.

Drosophila strains must be maintained by the frequent transfer of adult flies to new vials. This carries a danger of mutational deterioration and phenotypic changes. Development of an alternative method for long-term preservation without such changes is therefore imperative. Despite previous successful attempts, cryopreservation of Drosophila embryos is still not of practical use because of low reproducibility. Here, we describe a protocol for primordial germ cell (PGC) cryopreservation and strain revival via transplantation of cryopreserved PGCs into agametic Drosophila melanogaster (D. melanogaster) host embryos. PGCs are highly permeable to cryoprotective agents (CPAs), and developmental and morphological variation among strains is less problematic than in embryo cryopreservation. In this method, PGCs are collected from approximately 30 donor embryos, loaded into a needle after CPA treatment, and then cryopreserved in liquid nitrogen. To produce donor-derived gametes, the cryopreserved PGCs in a needle are thawed and then deposited into approximately 15agametic host embryos. A frequency of at least 15% fertile flies was achieved with this protocol, and the number of progeny per fertile couple was always more than enough to revive the original strain (the average progeny number being 77.2 ± 7.1), indicating the ability of cryopreserved PGCs to become germline stem cells. The average number of fertile flies per needle was 1.1 ± 0.2, and 9 out of 26 needles produced two or more fertile progeny. It was found that 11 needles are enough to produce 6 or more progeny, in which at least one female and one male are likely included. The agametic host makes it possible to revive the strain quickly by simply crossing newly emerged female and male flies. In addition, PGCs have the potential to be used in genetic engineering applications, such as genome editing.

Sonication results in variable quality and enhanced sensory attributes of Adajamir (Citrus assamensis) juice: A study on an underutilized fruit

Citrus assamensis, commonly known as Adajamir, is an underutilized fruit with distinctive sensory and nutritional properties. The limited amount of research on this particular citrus type was recognized as one of the research gaps for this study. The objective of this study was to evaluate and compare the impacts of sonication, pasteurization, and thermosonication techniques on the quality and sensory attributes of Adajamir juice. A randomized experimental design was used in the study, wherein the juice underwent three different treatments. The results indicate that there were no significant changes in pH or titratable acidity following all treatments. Yet, notable differences in juice color were observed. The use of sonication and thermosonication resulted in an increase in β-carotenoid levels. Additionally, total phenolic content and antioxidant activities were observed to increase. All three treatments led to a reduction in ascorbic acid levels relative to the control. However, the complete elimination of microbial growth was observed during the thermal treatment. Compared to other approaches, sonication has been shown to be notably more efficacious in enhancing both the flavor and aroma. Sonication has been observed to improve the perceived bitterness to a certain degree. These findings support the potential of sonication as an alternative preservation method for Adajamir juice, offering enhanced quality and sensory acceptance.

INFLUENCE OF THE HEATING AGENT TEMPERATURE ON THE KINETICS OF THE CONVECTIVE DRYING PROCESS AND THE CONTENT OF BIOACTIVE COMPOUNDS IN APPLE POMACE

An environmentally ecological approach to the reuse of agro-industrial products in the food industry involves extracting biologically active substances or using them as supplementary products rich in dietary fibers. Apple pomace obtained after juice extraction presents a product rich in biologically active substances, soluble and insoluble fibers, organic acids, and minerals. Convective drying of apple pomace is an alternative method for long-term preservation by maintaining the biologically active substances. The research aimed to study the influence of temperature in the range of 60-80 °C on the drying of apple pomace to a final moisture content of 12.0±0.13% using the convective method, and the effect of temperature on the yield of bioactive compounds and antioxidant activity in Gold Delicious apple pomace. Changing the air temperature during convective drying from 60 to 80 °C reduced the drying time by 1.6 times and demonstrated a 1.4-fold increase in drying rate. The increase in the thermal agent's temperature led to an increase in the kinetic characteristic values; KI values increased by 4%, and KII values increased by 44%. Spectrophotometric analysis demonstrated that apple pomace dried at a temperature of 70 °C had the highest total content of polyphenols, tannins, and carotenoids. Thus, the convective drying method of apple pomace in the temperature range of 60-80 °C exerts a significant influence on the composition of biologically active compounds in the apple pomace composition.

Developing A Herbal Preservative For Cadavers In Ayurvedic Anatomy- A Pilot Study

Background: Preserving cadavers is crucial for anatomical studies, but traditional methods using chemicalssuch as formalin can have harmful properties. Ayurveda, like other sciences, lacks an alternative method forpreservation. This study aimed to find a safe and effective herbal preservative for cadavers.Aim: The aim of this study was to formulate an herbal preservative solution for human cadavers that ensuresno risk of infection on contact, preservation of the body, and prevention of putrefaction changes andcontamination with maggots and insects.Objective: The objectives of the study were to identify natural preservative ingredients used in Ayurveda,research individual ingredients for their possible role as preservatives, and prepare and test an herbal solutionon chicken muscle pieces.Material and Methods: A detailed literature survey was conducted to identify natural preservative ingredientsused in Ayurveda. The individual ingredients were researched for their possible role as preservatives. An herbalsolution was prepared using the identified ingredients, and its efficacy was tested on chicken muscle pieces.Results: The study identified natural preservative ingredients such as Amalaka, Vibhitaki, Haritaki, Neembaand Vidanga. These ingredients were found to have antimicrobial, antioxidant, and anti-inflammatoryproperties that could potentially preserve human cadavers. Tested herbal preservative solution on chickenmuscle pieces at different concentrations. Preservation duration increased with higher concentration ofsolution. Maximum preservation period was achieved with 100% solution with no signs of contamination withmaggots and insects.Conclusion: This study provides a potential alternative to traditional chemical embalming methods and mayenable Ayurvedic graduates to successfully practice medicine and surgery. To validate the effectiveness of theherbal solution on human cadavers, additional research is required. Moreover, in the main study, it isrecommended to substitute certain drugs that could potentially enhance tissue preservation longevity in thissolution.

Using alternative preservation method in cryo-EM to overcome challenging specimen methionine tRNA synthetase

Targeting the protein machinery of intracellular parasites protein machinery is a challenge for drug development because often those parasites have minimal genomes and depend on their host for basic metabolic functions. Mycoplasma penetrans is one such intracellular parasite that infects immunocompromised patients, in particular individuals who are infected with HIV. Despite its minimal genome, the M. penetrans methionine tRNA synthetase (MetRS) is a unique fusion of an N-terminal alanine glyoxylate aminotransferase (AGAT) domain, which forms a homodimer, and a C-terminal tRS domain, the two of which are presumed to be independent. The AGAT domain appears to increase the MetRS enzyme's substrate differentiation ability and its efficiency in amino-acylating tRNA. The structural basis for this added functionality is unknown. We have now used cryo-EM to determine the structure of this unique drug target to show that the dimer adopts a butterfly-shaped arrangement, in which the body is the AGAT dimer, and the wings are two independent tRS domains. Initial cryogenic preparations of the complex were unsuccessful, so we also used a systematic analysis of alternative sample preparation techniques to identify which was optimal for this particular protein. With the structure in hand, we will next use rational mutagenesis to test the hypothesis that substrate channeling occurs amongst the unrelated active sites to impact MetRS activity, further developing this enzyme as a possible drug target.

The Impact of Plasma Activated Water Treatment on the Phenolic Profile, Vitamins Content, Antioxidant and Enzymatic Activities of Rocket-Salad Leaves.

Plasma activated water (PAW) recently received much attention as an alternative food preservation method. However, its effects on food quality are still scarce. This study evaluates the effect of PAW processing time on bioactive compounds of rocket-salad leaves including: 18 phenolic compounds, vitamin C, riboflavin, nicotinic acid, and nicotinamide. Moreover, the impact of PAW on both antioxidant (DPPH) and peroxidase (POD) activities was also investigated. This was performed using HPLC-DAD, HPLC-MS/MS, and spectrophotometric analysis. All treatments induced non-significant increases in total phenolic contents. However, depending on processing time, significant increases or decreases of individual phenolic compounds were observed. PAW-10 and -20 increased the ascorbic acid content to 382.76 and 363.14 mg/100 g, respectively, compared to control (337.73 mg/100 g). Riboflavin and nicotinic acid contents were increased significantly in PAW-20 (0.53 and 1.26 mg/100), compared to control (0.32 and 0.61 mg/100 g, respectively). However, nicotinamide showed non-significant increase in all treatments. Antioxidant activity improved significantly only in PAW-20, while peroxidase activity was reduced up to 36% in the longest treatment. In conclusion, PAW treatment could be an effective technique for rocket decontamination since it positively influenced the quality of rocket, improving the retention of polyphenols and vitamins.

Effects of freeze-drying on the quality and fertilising ability of goat sperm recovered from different parts of the epididymis

Lyophilisation is an alternative method for sperm preservation. The aim of this study was to evaluate the effects of freeze-thawing (F/T) and freeze-drying (F/D) on the quality of epididymal goat sperm. Sperm from each region of the epididymis (caput, corpus and cauda) were collected and evaluated for the expression of phospholipase C zeta (PLC-ζ), protamine 1 (PRM1), transition protein 1 (TNP1) and 2 (TNP2). The effects of F/T and F/D on sperm quality in terms of PLC-ζ expression, chromatin stability (Chromomycin A3; CMA3) and DNA integrity were examined. The fertilising ability after intracytoplasmic sperm injection (ICSI) was also tested. Fresh sperm existed PLC-ζ, PRM1, TNP1 and TNP2, irrespective of the regions of the epididymis. However, different patterns of PLC-ζ expression were found. Although PRM1, TNP1, TNP2 were still expressed after F/T or F/D, only F/T could preserve the presence of PLC-ζ. For fresh sperm, caput epididymal sperm had the lowest evidence of chromatin stability when compared to sperm harvested from other regions of the epididymis. The F/T and F/D further increased the numbers of CMA3-positive sperm (P < 0.001). In all cases, no CMA3 staining was observed in caudal epididymal sperm. The caudal epididymal sperm had significantly greater proportions of sperm with intact DNA compared with caput and corpus epididymal sperm, especially when F/T and F/D were performed. The fertilisation rates of F/D sperm tended to decrease when compared with F/T sperm (4.2 ± 3.2 vs. 13.6 ± 9.0, P = 0.08). It is concluded that the sperm recovered from the caudal epididymis is suitable for freezing and lyophilisation. However, poor fertilisation rates of F/D sperm were coincidently observed, with a deficit demonstration of PLC-ζ.

Comparing Life Cycle Assessment of Fish Preservation Method on Fishing Vessels in Indonesia

The economic benefit that the fishermen can obtain primarily depends on the quality of their catches. Therefore, it is essential to ensure that the caught fish stays fresh until entering the market. The most common fish preservation on traditional fishing boats in Indonesia is block ice, which must be carried in the fish hold from the home port. In comparison, another alternative method for fish preservation is the Flake Ice machine that can produce ice on-board. The study on Life Cycle Assessment (LCA) was conducted on these two fish preservation methods to understand the environmental impact of each technique. For undertaking the assessments, LCA software known as SimaPro was employed. The system boundary for the LCA about fish preservation applied the “well to wheels” approach and covered the whole service life of the vessel for the full consecutive 20 years of operation. The functional unit for the assessment was fish preservation of 4 tonnes during a 7-day fishing trip. The input of the evaluation was the materials and energy required to produce the amount of ice or to operate a refrigeration system for each year of the vessel’s operation, with a total of 30 trips per year. The paper explored the environmental impact of these two fish preservation methods in terms of Ozone Layer Depletion and Global Warming.