Preservation Protocols Research Articles

A novel method for the preparation of reproducible, stable, and non-infectious quality control materials for Chlamydia trachomatis nucleic acid detection.

Chlamydia trachomatis (CT) is a significant sexually transmitted pathogen known to evoke severe complications, including infertility. Nucleic acid amplification tests (NAATs) are recommended by the World Health Organization to detect CT infection. Furthermore, the establishment of methods, performance validation, internal quality control, and external quality assessment for CT NAATs necessitate the utilization of quality control materials (QCs). QCs are specimens or solutions that are analyzed for quality control purposes in a test system. In this study, we established a novel cell line that stably integrates CT amplification target sequences for producing QCs for CT NAATs. Utilizing clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 technology, we integrated the CT plasmid-mediated sequence (comprising the full length of the cryptic plasmid and the major outer membrane protein gene, 9,136 bp) into the MUC4 gene of HEK293T cells. Positive clones were screened through flow cytometric sorting, single-cell culture, and PCR-based identification, followed by the establishment of stable cell lines. These cells were then processed using optimized cell preservation procedures to prepare QCs. The sequence insertion copy number was confirmed by real-time quantitative PCR. This novel CT QCs demonstrate excellent clinical applicability, non-infectiousness, quantifiability, and stability. With an integrated sequence exceeding 9 kb in length, it offers exceptional flexibility for adapting to new kit developments. Furthermore, maintaining a well-defined copy number and stable shelf life, the QCs closely aligns with the quality control requirements of CT NAATs. This study presents an innovative method for preparing QCs for CT nucleic acid detection, making a valuable contribution to improving the performance of CT NAATs.IMPORTANCEUntreated CT infections impose significant burdens on individuals and communities, underscoring the importance of early and accurate testing via CT NAATs for disease control. QCs are instrumental in identifying testing process issues. Hence, we developed a cell line integrating CT-amplified target sequences as readily accessible non-infectious QCs. These QCs boast several advantages: the integration of over 9 kb of CT sequence allows for broad applicability, allowing flexible adaptation to the development of new kits. Confirming the CT sequence copy number provides a reliable basis for QC concentration preparation and kit detection limit evaluation. Optimized preservation protocol enhances QC stability during storage, facilitating convenient shipment to clinical laboratories at ambient temperatures. In summary, our novel CT QCs offer a powerful tool for improving CT NAAT performance and present a fresh perspective on QC preparation for detecting nucleic acids from intracellular parasitic pathogens.

A preclinical model for osteoarticular fracture fragment preservation for delayed re-implantation

BackgroundOpen articular fractures often include contaminated, devascularized osteoarticular fragments that are critical for joint reconstruction. Definitive treatment is often delayed such that decontamination and preservation of critical fragments for joint reconstruction is highly desirable. To validate decontamination and preservation protocols for safe and effective preservation of osteoarticular fragments for re-implantation, a preclinical animal model for inducing type 3 open articular fractures with contaminated, devascularized osteoarticular fragments was developed and validated. Materials and methodsWith IACUC approval, purpose-bred hounds (n = 5) were humanely euthanized. Immediately following euthanasia, a penetrating captive bolt pistol with 1.25 grain cartridge centered on the cranial aspect of each distal humerus was discharged to create open fractures in 3 dogs (6 elbows). In 2 dogs, matched osteoarticular tissues from non-injured elbows (controls) were retrieved for comparison. Distal humerus, proximal radius, and proximal ulna osteoarticular fragments (n = 27) were immediately placed in Missouri Osteochondral Preservation System (MOPS) solution and stored at room temperature. Radiographic, chondrocyte viability, and quantitative microbial culture assessments were performed immediately (time-0) and at 7 and 14 days of storage. ResultsThis preclinical canine model reliably produced type 3 open distal humeral fractures characterized by devascularized and contaminated osteoarticular fracture fragments. All fragments produced extensive microbial growth through 14 days of storage. Without decontamination, viable chondrocyte density in the fragments decreased significantly within 7 days, likely attributable to the profound contamination. ConclusionThese data highlight the importance of developing a reliable method for point-of-care decontamination and preservation of osteoarticular fracture fragments for safe and effective reimplantation of articular fracture fragments for joint reconstruction.

Biostasis: A Roadmap for Research in Preservation and Potential Revival of Humans.

Human biostasis, the preservation of a human when all other contemporary options for extension of quality life are exhausted, offers the speculative potential for survival via continuation of life in the future. While provably reversible preservation, also known as suspended animation, is not yet possible for humans, the primary justification for contemporary biostasis is the preservation of the brain, which is broadly considered the seat of memories, personality, and identity. By preserving the information contained within the brain's structures, it may be possible to resuscitate a healthy whole individual using advanced future technologies. There are numerous challenges in biostasis, including inadequacies in current preservation techniques, methods to evaluate the quality of preservation, and potential future revival technologies. In this report, we describe a roadmap that attempts to delineate research directions that could improve the field of biostasis, focusing on optimizing preservation protocols and establishing metrics for querying preservation quality, as well as pre- and post-cardiac arrest factors, stabilization strategies, and methods for long-term preservation. We acknowledge the highly theoretical nature of future revival technologies and the importance of achieving high-fidelity brain preservation to maximize the potential of future repair technologies. We plan to update the research roadmap biennially. Our goal is to encourage multidisciplinary communication and collaboration in this field.

Sub-zero non-freezing of vascularized composite allografts in a rodent partial hindlimb model

Ischemia is a major limiting factor in Vascularized Composite Allotransplantation (VCA) as irreversible muscular injury can occur after as early as 4–6 h of static cold storage (SCS). Organ preservation technologies have led to the development of storage protocols extending rat liver ex vivo preservation up to 4 days. Development of such a protocol for VCAs has the added challenge of inherent ice nucleating factors of the graft, therefore, this study focused on developing a robust protocol for VCA supercooling. Rodent partial hindlimbs underwent subnormothermic machine perfusion (SNMP) with several loading solutions, followed by a storage solution with cryoprotective agents (CPA) developed for VCAs. Storage occurred in suspended animation for 24h and VCAs were recovered using SNMP with modified Steen. This study shows a robust VCA supercooling preservation protocol in a rodent model. Further optimization is expected to allow for its application in a transplantation model, which would be a breakthrough in the field of VCA preservation.

Seminal plasma removal for medium-term preservation of ram sperm at 5 °C

This study aimed to investigate if washing ram sperm from seminal plasma (SP) could be an effective tool to extend sperm lifespan in medium-term preservation in liquid form to optimize ovine artificial insemination protocols. To this end, in Experiment 1 SP was added to a sperm model without previous contact with this substance (ram epididymal sperm) at the beginning or the end of a 48-hour preservation protocol at 5 °C (n = 13). Sperm motility and kinetic parameters and sperm functionality in terms of sperm viability, apoptosis, mitochondrial activity and reacted acrosomes were assessed after 6 h of storage at 15 °C (standard liquid preservation method) and 24 and 48 h at 5 °C. Extended sperm showed better results after 48 h when stored in the absence than in the presence of SP in most sperm quality parameters. Moreover, the final SP supplementation of this experimental group resulted in the highest sperm motility and kinetic parameters, viability and mitochondrial activity. These results suggested that initial SP deprivation could be beneficial in a medium-term ram sperm preservation protocol in liquid form, as well as a final supplementation. Therefore, we conducted Experiment 2 to evaluate the effect of SP removal from freshly ejaculated ram semen under the same storage conditions as in Experiment 1 (n = 12). Surprisingly, SP withdrawal impaired sperm functionality, leading to increased apoptosis and decreased mitochondrial activity after 24 and 48 h at 5 °C. Conversely, SP supplementation at the end of the preservation protocol of the ejaculate processed as usual had a positive effect on sperm quality and fertility. To summarize, SP absence was beneficial for a medium-term preservation protocol (up to 48 h at 5 °C) of ram epididymal sperm, but the same preservation protocol for ram ejaculated sperm revealed a possible failure of the SP removal method in avoiding the sperm-SP interaction effect. Meanwhile, SP supplementation of ram semen at the end of the preservation protocol increased in vitro sperm quality and fertility after artificial insemination.

Use of tamoxifene-controlled ovarian hyperstimulation for fertility preservation before breast cancer treatment: A prospective cohort study with a 5-year follow-up

PurposeFertility issues are of great concern for young women undergoing treatment for breast cancer (BC). Fertility preservation (FP) protocols using controlled ovarian stimulation (COS) with letrozole have been widely used with overall good results. However, letrozole cannot be used in every country in this context. This study aimed to assess the efficacy of tamoxifen for COS in women with early BC undergoing FP. MethodsThis multicentric prospective study included patients aged 18–40, diagnosed with stage I, II and III invasive BC, undergoing tamoxifen-COS before adjuvant or neoadjuvant chemotherapy (NAC). The primary endpoint was the efficacy of tamoxifen-COS protocol evaluated by the number of oocytes collected and vitrified. Secondary endpoints included the time interval before chemotherapy, breast cancer (BC) recurrence rates, and reproductive outcomes. ResultsNinety-five patients were included between 2014 and 2017, aged 31.5 ± 4 years on average. 37.9 % received NAC and 62.1 % received adjuvant chemotherapy. FP procedure was successful in 89.5 % of the cycles. The mean number of collected and vitrified oocytes was 12.8 ± 7.9 and 9.8 ± 6.2, respectively. The mean duration of COS was 10.4 ± 1.9 days. Median time before chemotherapy initiation was 3.6 weeks (IQR 3.1; 4.1) for women receiving NAC. Five-year relapse-free and overall survival rates were in-line with those expected in this population. Twenty-one women had spontaneous full-term pregnancies, while 5 underwent IVF cycles with frozen-thawed oocytes, without pregnancy. ConclusionTamoxifen-COS protocols appear to be feasible before adjuvant or NAC treatment in young BC patients and efficient in terms of oocyte yield.

Optimization of Hypothermic Protocols for Neurocognitive Preservation in Aortic Arch Surgery: A Literature Review.

Shifts from deep to moderate hypothermic circulatory arrest (HCA) in aortic arch surgery necessitate an examination of their differential impacts on neurocognitive functions, especially structured verbal memory, given its significance for patient recovery and quality of life. This study evaluates and synthesizes evidence on the effects of deep (≤20.0 °C), low-moderate (20.1-24.0 °C), and high-moderate (24.1-28.0 °C) hypothermic temperatures on structured verbal memory preservation and overall cognitive health in patients undergoing aortic arch surgery. We evaluated the latest literature from major medical databases such as PubMed and Scopus, focusing on research from 2020 to 2024, to gather comprehensive insights into the current landscape of temperature management during HCA. This comparative analysis highlights the viability of moderate hypothermia (20.1-28.0 °C), supported by recent trials and observational studies, as a method to achieve comparable neuroprotection with fewer complications than traditional deep hypothermia. Notably, low-moderate and high-moderate temperatures have been shown to support substantial survival rates, with impacts on structured verbal memory preservation that necessitate careful selection based on individual surgical risks and patient profiles. The findings advocate for a nuanced approach to selecting hypothermic protocols in aortic arch surgeries, emphasizing the importance of tailoring temperature management to optimize neurocognitive outcomes and patient recovery. This study fills a critical gap in the literature by providing evidence-based recommendations for temperature ranges during HCA, calling for ongoing updates to clinical guidelines and further research to refine these recommendations. The implications of temperature on survival rates, complications, and success rates underpin the necessity for evolving cardiopulmonary bypass techniques and cerebral perfusion strategies to enhance patient outcomes in complex cardiovascular procedures.

Hypothermic oxygenated perfusion of the donor heart in heart transplantation: the short-term outcome from a randomised, controlled, open-label, multicentre clinical trial

Static cold storage (SCS) remains the gold standard for preserving donor hearts before transplantation but is associated with ischaemia, anaerobic metabolism, and organ injuries, leading to patient morbidity and mortality. We aimed to evaluate whether continuous, hypothermic oxygenated machine perfusion (HOPE) of the donor heart is safe and superior compared with SCS. We performed a multinational, multicentre, randomised, controlled, open-label clinical trial with a superiority design at 15 transplant centres across eight European countries. Adult candidates for heart transplantation were eligible and randomly assigned in a 1:1 ratio. Donor inclusion criteria were age 18-70 years with no previous sternotomy and donation after brain death. In the treatment group, the preservation protocol involved the use of a portable machine perfusion system ensuring HOPE of the resting donor heart. The donor hearts in the control group underwent ischaemic SCS according to standard practices. The primary outcome was time to first event of a composite of either cardiac-related death, moderate or severe primary graft dysfunction (PGD) of the left ventricle, PGD of the right ventricle, acute cellular rejection at least grade 2R, or graft failure (with use of mechanical circulatory support or re-transplantation) within 30 days after transplantation. We included all patients who were randomly assigned, fulfilled inclusion and exclusion criteria, and received a transplant in the primary analysis and all patients who were randomly assigned and received a transplant in the safety analyses. This trial was registered with ClicalTrials.gov (NCT03991923) and is ongoing. A total of 229 patients were enrolled between Nov 25, 2020, and May 19, 2023. The primary analysis population included 204 patients who received a transplant. There were no patients who received a transplant lost to follow-up. All 100 donor hearts preserved with HOPE were transplantable after perfusion. The primary endpoint was registered in 19 (19%) of 101 patients in the HOPE group and 31 (30%) of 103 patients in the SCS group, corresponding to a risk reduction of 44% (hazard ratio 0·56; 95% CI 0·32-0·99; log-rank test p=0·059). PGD was the primary outcome event in 11 (11%) patients in the HOPE group and 29 (28%) in the SCS group (risk ratio 0·39; 95% CI 0·20-0·73). In the HOPE group, 63 (65%) patients had a reported serious adverse event (158 events) versus 87 (70%; 222 events) in the SCS group. Major adverse cardiac transplant events were reported in 18 (18%) and 33 (32%) patients in the HOPE and SCS group (risk ratio 0·56; 95% CI 0·34-0·92). Although there was not a significant difference in the primary endpoint, the 44% risk reduction associated with HOPE was suggested to be a clinically meaningful benefit. Post-transplant complications, measured as major adverse cardiac transplant events, were reduced. Analysis of secondary outcomes suggested that HOPE was beneficial in reducing primary graft dysfunction. HOPE in donor heart preservation addresses the existing challenges associated with graft preservation and the increasing complexity of donors and heart transplantation recipients. Future investigation will help to further elucidate the benefit of HOPE. XVIVO Perfusion.

261 High rate of Dropping from follow-up hinders implementing rectal cancer organ preservation protocols

261 High rate of Dropping from follow-up hinders implementing rectal cancer organ preservation protocols

The Hypogeous Roman Archeological Museum of Positano: Study of the Evolution of Biological Threaten and Development of Adequate Control Protocols.

Hypogea are natural or artificial spaces located underground often of great interest from an anthropological, archeological, religious, artistic, or historic point of view. Due to their features, these environments usually present conservative problems and biological colonization could be considered as one of the main threats. The present three-year study was carried out by specialists of the Central Institute for Restoration of Rome (ICR) in the hypogeous site preserved in the Roman Archeological Museum of Positano (Positano MAR) and focused on characterizing biological alterations present on the mural paintings; setting up efficient strategies and protocols for biodeterioration control; and monitoring the efficacy of direct and indirect interventions. Patinas with different morphologies were analyzed through microscopic observations, cultural analyses and next-generation sequencing. The results proved that the alterations comprised a great variety of microorganisms forming very distinct communities, differently distributed over space and time. The main taxa represented were bacteria of phyla Pseudomonadota and Actinomycetota, fungi belonging to the genus Fusarium and Gliocladium, and algae of the genus Chlorococcum. Preservation protocols were set up considering the alterations' composition and included the application of biocides, limiting daily temperature changes, decreasing illuminance values on painted surfaces, and the screening of natural light sources.

Validation of a forensic evidence collection and preservation protocol in emergency settings: Delphi technique

Validation of a forensic evidence collection and preservation protocol in emergency settings: Delphi technique

Smart city energy efficient data privacy preservation protocol based on biometrics and fuzzy commitment scheme

Advancements in cloud computing, flying ad-hoc networks, wireless sensor networks, artificial intelligence, big data, 5th generation mobile network and internet of things have led to the development of smart cities. Owing to their massive interconnectedness, high volumes of data are collected and exchanged over the public internet. Therefore, the exchanged messages are susceptible to numerous security and privacy threats across these open public channels. Although many security techniques have been designed to address this issue, most of them are still vulnerable to attacks while some deploy computationally extensive cryptographic operations such as bilinear pairings and blockchain. In this paper, we leverage on biometrics, error correction codes and fuzzy commitment schemes to develop a secure and energy efficient authentication scheme for the smart cities. This is informed by the fact that biometric data is cumbersome to reproduce and hence attacks such as side-channeling are thwarted. We formally analyze the security of our protocol using the Burrows–Abadi–Needham logic logic, which shows that our scheme achieves strong mutual authentication among the communicating entities. The semantic analysis of our protocol shows that it mitigates attacks such as de-synchronization, eavesdropping, session hijacking, forgery and side-channeling. In addition, its formal security analysis demonstrates that it is secure under the Canetti and Krawczyk attack model. In terms of performance, our scheme is shown to reduce the computation overheads by 20.7% and hence is the most efficient among the state-of-the-art protocols.

Poster 296: Comparing Outcomes of Osteochondral Allograft Transplantation of the Knee Preserved by the Missouri Osteochondral Preservation System to Standard Preservation Methods

Objectives: Osteochondral allograft (OCA) transplantation is an established surgical intervention for full-thickness chondral defects. Standard Preservation (SP) protocols currently allow for osteochondral allografts to be stored for a maximum of 28 days after procurement. The Missouri Osteochondral Allograft Preservation System allows for grafts to be stored for at least 60 days after procurement. The purpose of this study is to compare clinical and radiographic outcomes in patients that have undergone OCA transplantation utilizing Missouri Osteochondral Allograft Preservation System (MOPS) to standard preservation protocols. Methods: Retrospective review identified all patients who underwent OCA transplantation between 2012 and 2018. Patients without appropriate follow-up, postoperative outcomes, or necessary imaging were excluded. All patients had at least 2-year follow-up data. Patient demographics, outcomes, and graft information were compared between patients who received grafts preserved by MOPS and standard preservation as previously defined. Results: Sixty-four patients were included. Twenty-five were in the MOPS group and 39 in the SP group. Members of the MOPS group were younger at the time of surgery than those in the SP group (28.6 ± 10.7 years in MOPS vs. 36.7 ± 10.9 in SP group, P = 0.005). Seventy-six grafts were analyzed with 39 grafts in the MOPS group and 37 in the SP group. Grafts in the MOPS group were stored longer (average 50.03 days) than grafts in the SP group (average 23.18 days) (P < 0.001). No difference between groups was found in complications or reoperation rates. Outcomes between the 2 groups showed no statistical differences for complications or revisions as well as similar Knee Injury and Osteoarthritis Outcome Scores for Joint Replacement, International Knee Documentation Committee scores, 12-item psychosocial safety climate scores, and 12-item mental health component summary scores. Conclusions: Grafts preserved using the MOPS were stored for more than double the length of time than grafts preserved with standard preservation techniques. There was no difference in postoperative outcomes following OCA transplantation regardless of preservation method used.

Emerging importance of stool preservation methods in OMICS studies with special focus on cancer biology.

The intricate consortium of microorganisms in the human gut plays a crucial role in different physiological functions. The complex known-unknown elements of the gut microbiome are perplexing and the absence of standardized procedures for collecting and preserving samples has hindered continuous research in comprehending it. The technological bias produced because of lack of standard protocols has affected the reproducibility of results. The complex nature of diseases like colorectal cancer, gastric cancer, hepatocellular carcinoma and breast cancer require a thorough understanding of its etiology for an efficient and timely diagnosis. The designated protocols for collection and preservation of stool specimens have great variance, hence generate inconsistencies in OMICS studies. Due to the complications associated to the nature of sample, it is important to preserve the sample to be studied later in a laboratory or to be used in the future research purpose. Stool preservation is gaining importance due to the increased use of treatment options like fecal microbiota transplantation to cure conditions like recurrent Clostridium difficile infections and for OMICS studies including metagenomics, metabolomics and culturomics. This review provides an insight into the importance of omics studies for the identification and development of novel biomarkers for quick and noninvasive diagnosis of various diseases.

Upfront Neck Dissection in Organ Preservation Protocol in Head-Neck SCC : Can it be a Game Changer?

The major prognostic factor in head-neck squamous cell carcinoma is the status of lymph nodes. Though there are studies addressing the role of neck dissection in organ preservation protocol, a lacuna still exists as far as the question of upfront neck dissection arises. Despite the potential benefits of upfront neck dissection; the benefits have not been well researched and thus its place in management of head-neck cancers undergoing organ preservation protocol is still questionable. We conducted an observational study in 22 patients with T1 - 2N2 - 3a biopsy proven squamous cell carcinoma of hypopharynx, oropharynx and laryngopharynx which were eligible for organ preservation protocol for their primary site. The primary outcome was to calculate their overall survival rate at 5 years. Overall 5 year survival rate was 68.12%, 5-year Disease free survival rate was 77.2%. The time to start for the primary site, after neck dissection however had a significant effect on overall survival. Those who had a delay of 21 days or less had 80.1% survival at 5 years compared to those who started at 30 days (61.3%) and drastically reduced in those who started at 39 days (20.4%). Log-rank test for the effect of nodal status showed a significant difference in terms of 5 year survival between the groups(p = .027). The 5 year survival rates for N2A, N3A and N2B nodal status were 88.8%, 50% ,and 54.5% ,respectively. We strongly favour upfront neck dissections in a subset of head-neck squamous cell carcinoma cases with T1 - 2N2 - 3a eligible for organ preservation regimen.

Update of the cooling protocol for antibiotic-free storage of boar semen at 5°C improves sperm quality and maintains low bacterial counts.

Preserving boar semen at 5°C instead of the conventional storage temperature of 17°C would enable a reduction of antibiotic use in pig insemination. To protect the chilling-sensitive boar spermatozoa, holding the extended semen at a higher temperature before cooling could be beneficial and facilitate the implementation of the innovative preservation concept in practice, provided that bacterial growth is kept at a low level. The aim of this study was to introduce a holding time (HT) at 17°C before cooling and to examine the effect on sperm quality and bacterial growth compared to the original cooling protocol for antibiotic-free 5°C semen storage. A series of experiments with semen doses from eight boars extended in Androstar® Premium without conventional antibiotics revealed that sperm kinematics and the integrity of sperm plasma membranes and acrosomes were improved with HT between 16 and 24 h followed by delayed cooling with 0.04°C/min when compared to the original protocol for semen preservation at 5°C (p < 0.05). Both a shorter HT of 6 h and a faster cooling rate of 0.07°C/min reduced sperm quality (p < 0.05). The HT for 24 h did not compromise the inhibitory effect on bacterial growth during long-term semen storage at 5°C, not even in semen doses spiked with Serratia marcescens. In conclusion, semen storage at 5°C with the modified cooling protocol improved sperm quality and is antimicrobially efficient. It thus presents a ready-to-use tool for a reduction or replacement of antibiotics in pig insemination.

One hundred fifty years of total laryngectomies.

The inaugural total laryngectomy in history was conducted by Billroth in 1873. Nevertheless, significant enhancements to the technique were achieved through the contributions of Gluck, Sorensen, and various other surgeons. Throughout the twentieth century, advancements in anesthesia, infectious disease, hospital hygiene, antibiotic therapy, resuscitation, and the expertise of numerous laryngologists elevated total laryngectomy to a pivotal surgical intervention in head and neck surgery. The latter half of the twentieth century witnessed a paradigm shift with the emergence of organ preservation protocols. Total laryngectomy became the preferred choice for patients experiencing radiotherapy failure. However, the widespread use of laryngeal conservative treatments appears to be correlated with a decline in overall survival rates in the United States and Europe. The evolution of new minimally invasive surgical approaches in the twenty-first century may usher in a revolutionary era in the management of laryngeal carcinoma, offering the potential for improved survival and functional outcomes.

Short- and long term preservation of cardiac tissue-engineered constructs

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – EU funding. Main funding source(s): This research was financially supported by the Gravitation Program ‘‘Materials Driven Regeneration,’’ funded by the Netherlands Organization for Scientific Research (024.003.013) and the EU-funded project BRAV∃ (H2020, ID:874827). Background Cardiac tissue engineering (cTE) has already advanced towards the first clinical trials, investigating safety and feasibility of cTE construct transplantation in failing hearts. However, the lack of well-established preservation methods poses a hindrance to further scalability, commercialization, and transportation, thereby reducing their clinical implementation. Purpose In this study, hypothermic (short-term) preservation (4°C), and slow and fast freezing methods for cryopreservation, at -196°C and -150°C respectively, were investigated as potential solutions to extend the cTE construct implantation window and increase their clinical applicability. Material and Methods The cTE model used consisted of induced-pluripotent stem cell-derived cardiomyocytes and human cardiac fibroblasts embedded in a natural-derived hydrogel and supported by a polymeric melt electrowritten hexagonal scaffold. Constructs, composed of cardiomyocytes of different maturity, were preserved for three days, using several commercially available preservation protocols and solutions. Their viability, function (BPM and calcium handling), and metabolic activity were investigated after rewarming. Results Our observations suggest that cardiomyocyte maturation did not influence post-rewarming viability, however, aging positively influenced construct function pre- and post-storage. Hypothermic preservation in Hypothermosol ensured retained construct viability and function. Furthermore, fast-freezing outperformed slow-freezing, but both viability and function were severely reduced after rewarming. Conclusion In conclusion, whereas long-term preservation remains a challenge, hypothermic preservation in Hypothermosol represents a promising solution for cTE construct short-term preservation and potential transportation, aiding in off-the-shelf availability, ultimately increasing their clinical applicability.Graphical abstract

Orthopedic Joint Preservation: A Comprehensive Review

This comprehensive review article delves into the intricate realm of orthopedic joint preservation with a focus on surgical techniques, innovative treatments, rehabilitation protocols, and long-term outcomes. Within the orthopedic landscape, joint preservation plays a crucial role in enhancing patient quality of life and functional outcomes. The article delivers a detailed exploration of various surgical techniques utilized for joint preservation, encompassing arthroscopy, cartilage repair, osteotomy, and ligament reconstruction. By scrutinizing the indications, outcomes, and potential complications associated with these procedures, the article aids in providing a comprehensive understanding for practitioners. Moreover, it delves into the realm of cutting-edge treatments such as regenerative medicine, stem cell therapy, and biologic agents, shedding light on their significant role in promoting joint preservation and fostering tissue healing. Rehabilitation protocols for joint preservation are also thoroughly evaluated, underlining the pivotal role of early mobilization, tailored physical therapy regimens, and patient education in fostering optimal outcomes while averting potential complications. Through an assessment of long-term outcomes post joint preservation interventions including functional improvements, pain management, and patient contentment, the article endeavors to gauge the efficacy and longevity of these interventions. By offering a nuanced overview of the current state of joint preservation in orthopedics, the article extends valuable insights to orthopedic surgeons, researchers, and healthcare professionals actively engaged in managing joint disorders.

Ensuring Accuracy: Critical Validation Techniques in Geochemical Analysis for Sustainable Geothermal Energy Development

Geochemical analysis is a critical tool in geothermal exploration, providing valuable insights into reservoir characteristics. However, obtaining accurate and reliable geochemical data requires rigorous validation techniques. This review examines key factors affecting the accuracy of geochemical data and discusses best practices for ensuring quality. Proper sampling methods, including selection of representative locations, use of appropriate equipment, and adherence to robust protocols for sample collection, filtration, preservation, and storage, are essential for maintaining integrity. Analytical techniques must be carefully selected, with regular calibration and standardization of instruments using certified reference materials. Implementing comprehensive quality assurance and quality control procedures, such as analyzing blanks, duplicates, and spike samples, helps monitor precision and accuracy. Data interpretation should consider the complexities of the geological and hydrological settings, integrating multiple lines of evidence. By following established guidelines and continuously updating methods based on emerging technologies and inter-laboratory comparisons, geothermal teams can optimize the reliability of their geochemical data. Accurate and precise geochemical information, when combined with geological, geophysical, and hydrological data, enables informed decision-making and enhances the success of geothermal projects. As geothermal energy gains importance in the transition to sustainable resources, ensuring the accuracy of geochemical analysis will be crucial for effective exploration and development.