Expertise In Biology Research Articles

The development of molecular genetics concept test for senior high school students using Rasch analysis

<span lang="EN-US">Developing a high-quality test item requires substantial time and effort. A well-developed item bank is conducted using rigorous development and validation procedures. This study aimed to describe the development process of molecular genetics concept test (MGCT) for senior high school students using Rasch analysis under <a name="_Hlk107307570"></a>Berkeley evaluation and assessment research (BEAR) assessment system framework. The test consists of 50 multiple-choice items to assess conceptual understanding of molecular genetics concepts. The MGCT was developed based on curriculum analysis from the Indonesian ministry of education and culture and content-validated by three content experts comprising an expert in biology, an expert in bioinformatics, and an experienced Indonesian biology teacher in a senior high school. The MGCT was then piloted to 114 students who had taught the molecular genetics unit from a senior high school to conduct the empirical validation. The results from Rasch analysis showed that the MGCT is acceptable because all items have outfit and infit mean-square values in the acceptable range of 0.7 to 1.3 and the reliability is 0.43. So, the MGCT can be used to assess the understanding of the molecular genetics concept. However, several items were too difficult to discriminate the student ability. So, future studies need to develop more this MGCT to get a more appropriate instrument.</span>

A Personal Perspective on My Scientific Career

A Personal Perspective on My Scientific Career

Tracking the Travels

Tracking the Travels

Off the deep end: What can deep learning do for the gene expression field?

After a COVID-related hiatus, the fifth biennial symposium on Evolution and Core Processes in Gene Regulation met at the Stowers Institute in Kansas City, Missouri July 21 to 24, 2022. This symposium, sponsored by the American Society for Biochemistry and Molecular Biology (ASBMB), featured experts in gene regulation and evolutionary biology. Topic areas covered enhancer evolution, the cis-regulatory code, and regulatory variation, with an overall focus on bringing the power of deep learning (DL) to decipher DNA sequence information. DL is a machine learning method that uses neural networks to learn complex rules that make predictions about diverse types of data. When DL models are trained to predict genomic data from DNA sequence information, their high prediction accuracy allows the identification of impactful genetic variants within and across species. In addition, the learned sequence rules can be extracted from the model and provide important clues about the mechanistic underpinnings of the cis-regulatory code.

Cell scientist to watch – Marie Barberon

ABSTRACT Marie Barberon studied plant biology at the University of Montpellier II, France. She then stayed in Montpellier for her PhD with Grégory Vert to work on iron transport and signalling, focussing on the trafficking and functions of a metal transporter. Marie then moved to Lausanne, Switzerland, and received an EMBO long-term fellowship to carry out her postdoctoral work with Niko Geldner. There, she investigated the function of the endodermis as a barrier for nutrient acquisition. Marie joined the University of Geneva in February 2018 as Assistant Professor, where she is combining her expertise in nutrient physiology, cell biology and development to uncover the mechanisms controlling radial transport of nutrients in roots.

M6A-SAC-seq for quantitative whole transcriptome m6A profiling.

N6-methyladenosine (m6A) is the most abundant mRNA modification in mammalian cells, regulating many physiological processes. Here we describe a method for base-resolution, quantitative m6A sequencing in the whole transcriptome. The enzyme and small-molecule cofactor used in this protocol are prepared by recombinant protein expression and organic synthesis, respectively. Then the library can be prepared from various types of RNA samples using a ligation-based strategy, with m6A modifications being labeled by the enzyme and cofactor. Detailed instructions on ensuing data analysis are also included in this protocol. The method generates highly reproducible results, uncovering 31,233-129,263 sites using as little as 2 ng of poly A+ RNA. These identified sites correspond well with previous m6A profiling results, covering over 65% of peaks detected by the antibody-based approaches. Compared with other currently available methods, this method can be applied to various types of biological samples, including fresh and frozen tissues as well as formalin-fixed paraffin-embedded samples, providing a quantitative method to uncover new insights into m6A biology. The protocol requires basic expertise in molecular biology, recombinant protein expression and organic synthesis. The whole protocol can be done in 15 days, with the library preparation taking 5 days.

ЛЕВ АНАТОЛЬЕВИЧ ЖИВОТОВСКИЙ (к 80-летию со дня рождения)

November 22, 2022 is the 80th birthday of Lev Anatolyevich Zhivotovsky, an outstanding scientist, expert in population and mathematical biology, Professor, PhD in biology and mathematics, head of a laboratory at Research Institute of General Genetics of the Russia Academy of Sciences, principal researcher at All-Russia Research Institute of Fishery and Oceanography, Honored Scientist of the Russian Federation, laureate of Science and Technology Award of the Russian Federation, of I.I. Schmalhausen Evolutionary Biology Award of the Russian Academy of Sciences, and of The Lancet Award for the Best Article of the Year (2003), and just a wonderful personality. Most of his research focuses on studying natural populations of plants and animals. Jointly with his colleagues, he studied the population structure of oaks in the Caucasus, the genetics of conifers and of agricultural plants (cotton, grapes etc.) and addressed flora restoration in oil production areas in Tyumen Region. Based on these works, patents were obtained for methods of selection and breeding of plant populations. He introduced a new population indicator «efficiency index» and a new classification of normal plant populations («delta-omega»). Based on the concerted application of the efficiency index and the age index suggested by A.A. Uranov, Lev A. developed an ecogeographic approach by proposing the concept of ecogeographic units thereby making a significant contribution to the development of a general theory of species population structure. Lev A. Zhivitovsky has been being a scientific supervisor and participant of many expeditions to the Far East, Siberia and other regions of the Russian Federation. He has authored/coauthored more than 300 research articles and eight monographs, including his fundamental textbook «The Genetics of Natural Populations» (2021).

22. Reimagining and enhancing the Clinical Genome Resource (ClinGen) Somatic Cancer Clinical Domain Working Group

Interpretation of the clinical significance of somatic variants in cancer remains a major challenge in cancer diagnosis, prognosis, and treatment decisions. The Clinical Genome Resource (ClinGen) Somatic Cancer Clinical Domain Working Group (CDWG) facilitates the development of data curation guidelines and standards to determine the clinical significance of somatic alterations in cancer through structured collaboration. Over 180 multi-disciplinary experts in cancer biology, oncology, pathology, genetics, genomics, and informatics have come together to create high-quality, clinically-significant somatic cancer variant assertions in the CIViC knowledgebase, with further dissemination through other resources like ClinVar and the Variant Interpretation for Cancer Consortium (VICC) MetaKB. The Somatic CDWG has established the Pediatric Cancer and Hematological Cancer Taskforces, whose efforts facilitated membership growth and targeted curation projects. Their success led to the recent creation of the Solid Tumor Taskforce. Taskforces serve as incubators for the formation of Somatic Cancer Variant Curation Expert Panels (SC-VCEPs), the primary generators of the aforementioned high-level clinical assertions. SC-VCEPs undergo a 4-step approval process adapted from ClinGen germline VCEP procedures and develop granular specifications for their genomic alterations of interest through modifications of AMP/ASCO/CAP [PMID:27993330] and ClinGen/CGC/VICC Oncogenicity [PMID:35101336] guidelines. Three SC-VCEPs (FGFR variants, NTRK-fusions, and FLT3 variants) are working through this process and three (Histone H3, BCR::ABL1-like B-ALL, and Clinical Actionability) are in the planning stage. ClinGen Somatic groups have generated 634 CIViC Evidence Items and 21 assertions of clinical significance from 365 published papers. Our Taskforces and SC-VCEPs foster collaboration, innovation, and most importantly the advancement of precision medicine.

Process-Sensitive Naming: Trait Descriptors and the Shifting Semantics of Plant (Data) Science

This paper examines classification practices in the domain of plant data semantics, and particularly methods used to label plant traits to foster the collection, management, linkage and analysis of data about crops across locations—which crucially inform research and interventions on plants and agriculture. The efforts required to share data place in sharp relief the forms of diversity characterizing the systems used to capture the biological and environmental characteristics of plant variants: particularly the biological, cultural, scientific and semantic diversity affecting the identification and description of plant traits, the methods used to generate and process data, and the goals and skills of those with relevant expertise—including farmers and breeders. Through a study of the Crop Ontology (which explicitly recognizes and negotiates diversity) and its application to cassava breeding, I argue for a process-sensitive approach to the naming of plant traits that focuses on documenting environmental processes instead of biological products. I claim that this approach can foster reliable linkage and robust re-use of plant data, while at the same time facilitating dialogue between data scientists, plant researchers, breeders, and other relevant experts in ways that crucially inform agricultural interventions. I conclude that the study of data semantics and related descriptors constitutes a productive and underexplored way to think about the epistemic import of naming traits within plant science. The effort to articulate semantic differences among plant varieties and methods of data processing can generate newly inclusive ways to develop and communicate biological knowledge. In turn, such practices have the potential to defy existing understandings of systematisation and hierarchies of expertise in biology, thus bolstering the extent to which plant science can support biodiversity and sustainable agriculture.

Recent Advancements in Surface Modification, Characterization and Functionalization for Enhancing the Biocompatibility and Corrosion Resistance of Biomedical Implants

Metallic materials are among the most crucial engineering materials widely utilized as biomaterials owing to their significant thermal conductivity, mechanical characteristics, and biocompatibility. Although these metallic biomedical implants, such as stainless steel, gold, silver, dental amalgams, Co-Cr, and Ti alloys, are generally used for bone tissue regeneration and repairing bodily tissue, the need for innovative technologies is required owing to the sensitivity of medical applications and to avoid any potential harmful reactions, thereby improving the implant to bone integration and prohibiting infection lea by corrosion and excessive stress. Taking this into consideration, several research and developments in biomaterial surface modification are geared toward resolving these issues in bone-related medical therapies/implants offering a substantial influence on cell adherence, increasing the longevity of the implant and rejuvenation along with the expansion in cell and molecular biology expertise. The primary objective of this review is to reaffirm the significance of surface modification of biomedical implants by enlightening numerous significant physical surface modifications, including ultrasonic nanocrystal surface modification, thermal spraying, ion implantation, glow discharge plasma, electrophoretic deposition, and physical vapor deposition. Furthermore, we also focused on the characteristics of some commonly used biomedical alloys, such as stainless steel, Co-Cr, and Ti alloys.

Organoids as tools for fundamental discovery and translation-a Keystone Symposia report.

Complex three-dimensional in vitro organ-like models, or organoids, offer a unique biological tool with distinct advantages over two-dimensional cell culture systems, which can be too simplistic, and animal models, which can be too complex and may fail to recapitulate human physiology and pathology. Significant progress has been made in driving stem cells to differentiate into different organoid types, though several challenges remain. For example, many organoid models suffer from high heterogeneity, and it can be difficult to fully incorporate the complexity of in vivo tissue and organ development to faithfully reproduce human biology. Successfully addressing such limitations would increase the viability of organoids as models for drug development and preclinical testing. On April 3-6, 2022, experts in organoid development and biology convened at the Keystone Symposium "Organoids as Tools for Fundamental Discovery and Translation" to discuss recent advances and insights from this relatively new model system into human development and disease.

FluxomicsExplorer: Differential visual analysis of Flux Sampling based on Metabolomics

Flux Sampling (FS) is an unbiased constraint-based modeling method in systems biology that characterizes the admissible metabolic flux space of cells and organisms at a molecular level. It returns information about reaction flux ranges and their probability from a genome-scale metabolic model (GEM), which can be used to predict metabolic phenotypes. FS is particularly beneficial if a cellular objective cannot be easily identified. We present FluxomicsExplorer, a web-based visual analysis application that facilitates a data- and task-driven exploratory analysis of FS results. It was designed together with experts in medicine and systems biology to satisfy their requirements and offers multiple linked views that visualize the standardized scoring of flux differences, flux distributions, and the network of biological pathways to guide the analysis process. Instead of focusing on a subset of known influential reactions for the analysis, it allows users to assess the results as a whole concerning the underlying GEM. The visualizations guide the user to affected reactions, metabolites, and network areas by highlighting relevant changes between two conditions. These flux changes can then be analyzed in detail by visualizing the reaction flux distributions. We show FluxomicsExplorer’s utility using a case study of the red blood cell metabolism in different blood plasma environments.

MobileOG-db: a Manually Curated Database of Protein Families Mediating the Life Cycle of Bacterial Mobile Genetic Elements.

ABSTRACTBacterial mobile genetic elements (MGEs) encode functional modules that perform both core and accessory functions for the element, the latter of which are often only transiently associated with the element. The presence of these accessory genes, which are often close homologs to primarily immobile genes, incur high rates of false positives and, therefore, limits the usability of these databases for MGE annotation. To overcome this limitation, we analyzed 10,776,849 protein sequences derived from eight MGE databases to compile a comprehensive set of 6,140 manually curated protein families that are linked to the “life cycle” (integration/excision, replication/recombination/repair, transfer, stability/transfer/defense, and phage-specific processes) of plasmids, phages, integrative, transposable, and conjugative elements. We overlay experimental information where available to create a tiered annotation scheme of high-quality annotations and annotations inferred exclusively through bioinformatic evidence. We additionally provide an MGE-class label for each entry (e.g., plasmid or integrative element), and assign to each entry a major and minor category. The resulting database, mobileOG-db (for mobile orthologous groups), comprises over 700,000 deduplicated sequences encompassing five major mobileOG categories and more than 50 minor categories, providing a structured language and interpretable basis for an array of MGE-centered analyses. mobileOG-db can be accessed at mobileogdb.flsi.cloud.vt.edu/, where users can select, refine, and analyze custom subsets of the dynamic mobilome.IMPORTANCE The analysis of bacterial mobile genetic elements (MGEs) in genomic data is a critical step toward profiling the root causes of antibiotic resistance, phenotypic or metabolic diversity, and the evolution of bacterial genera. Existing methods for MGE annotation pose high barriers of biological and computational expertise to properly harness. To bridge this gap, we systematically analyzed 10,776,849 proteins derived from eight databases of MGEs to identify 6,140 MGE protein families that can serve as candidate hallmarks, i.e., proteins that can be used as “signatures” of MGEs to aid annotation. The resulting resource, mobileOG-db, provides a multilevel classification scheme that encompasses plasmid, phage, integrative, and transposable element protein families categorized into five major mobileOG categories and more than 50 minor categories. mobileOG-db thus provides a rich resource for simple and intuitive element annotation that can be integrated seamlessly into existing MGE detection pipelines and colocalization analyses.

Learning from the COVID-19 Pandemic Crisis to Overcome the Global Environmental Crisis

The COVID-19 crisis and the environmental crisis share a range of similarities. Both crises take place on a global scale and affect all aspects of our lives. However, we humans respond differently to these challenges. Here, we compare and comment on characteristics of the COVID-19 crisis and the environmental crisis, explore how far these two crises are comparable, and what we can learn from actions that have been taken against the COVID-19 crisis. We discuss how human societies are affected by the respective crises, and analyze policy makers’ responses and offer pathways to better inform policy. We highlight the role of science, which significantly contributed to decision making throughout the COVID-19 crisis, but seems frequently underrepresented in the environmental crisis. We conclude that there are significant differences between the two crises in terms of perceptibility and thus communicability. While problems and solutions in the COVID-19 crisis are largely linearly correlated, the challenges of the environmental crisis are far more complex and decoupled, and thus appear much more complex and are often only perceived with difficulty by humans. Thus, tackling the environmental crisis is much more challenging than solving the COVID-19 crisis. To overcome the environmental crisis, purely technical approaches for combating symptoms are not sufficient. However, political interests are usually short-term, and do not correspond with the temporal and spatial scales of global change. There is an urgent need to improve institutionalized scientific advisory mechanisms and to empower global policy makers who are independent of local interest groups. Furthermore, we need the sound communication of complex interactions to the general public and the translation of scientific findings into action. One possibility to achieve this is to bring together natural scientists with expertise in biology, climate and geosciences and social scientists, psychologists, and, possibly, artists.

Essential Conditions for the Full Synergy of Probability of Occurrence Distributions.

In this contribution, we specify the conditions for assuring the validity of the synergy of the distribution of probabilities of occurrence. We also study the subsequent restriction on the maximal extension of the strict concavity region on the parameter space of Sharma–Mittal entropy measures, which has been derived in a previous paper in this journal. The present paper is then a necessary complement to that publication. Some applications of the techniques introduced here are applied to protein domain families (Pfam databases, versions 27.0 and 35.0). The results will show evidence of their usefulness for testing the classification work performed with methods of alignment that are used by expert biologists.

Biology Students Association as the base for establishing future experts and cave fauna exploration

Biology Students Association – BIUS (BIUS) from Zagreb, Croatia is a non-government and non-profit association founded in 1999 by biology students from the Faculty of Science, University of Zagreb. Aiming to develop future experts in different fields of biology, BIUS is composed of groups that specialize in various subject areas. Besides holding diverse science lectures, workshops, and field trips, the main BIUS' activities are annual student research camps with the participation of up to 130 members. An integral part of BIUS from the very beginning is the Biospeleology group (Group). During these first 23 years of the Group's existence, many members discovered their interest in cave biology through field research, cabinet work, and educational workshops. The Group organized two student research camps and participated in 16 others. In addition, the Group organized seven independent projects. Throughout all of these projects, cave fauna was collected from over 150 caves and pits at different localities of Croatian karst. The Group's long-term existence is facilitated by the guidance of mentors from the Croatian Biospeleological Society. Altogether, the Group provides numerous skills and valuable opportunities to students and is important for developing future scientific experts in cave biology in Croatia. So far, 12 Group members have written their master thesis and four have their doctoral thesis in the field of cave biology. Former members currently work at universities, research institutes, government, and civil sector, or as entrepreneurs on topics closely related to cave biology and karst research.

The Core Human Microbiome: Does It Exist and How Can We Find It? A Critical Review of the Concept.

The core microbiome, which refers to a set of consistent microbial features across populations, is of major interest in microbiome research and has been addressed by numerous studies. Understanding the core microbiome can help identify elements that lead to dysbiosis, and lead to treatments for microbiome-related health states. However, defining the core microbiome is a complex task at several levels. In this review, we consider the current state of core human microbiome research. We consider the knowledge that has been gained, the factors limiting our ability to achieve a reliable description of the core human microbiome, and the fields most likely to improve that ability. DNA sequencing technologies and the methods for analyzing metagenomics and amplicon data will most likely facilitate higher accuracy and resolution in describing the microbiome. However, more effort should be invested in characterizing the microbiome’s interactions with its human host, including the immune system and nutrition. Other components of this holobiontic system should also be emphasized, such as fungi, protists, lower eukaryotes, viruses, and phages. Most importantly, a collaborative effort of experts in microbiology, nutrition, immunology, medicine, systems biology, bioinformatics, and machine learning is probably required to identify the traits of the core human microbiome.

Functionality and Clinical Effects of Anti-Cov2 Vaccines (Aka Mrna) And Integration on Mitochondrial DNA

1. This issue would require many pages, but it is briefly described here for easier understanding 2. The writer refers to the Molecular Biology Experts and to reading the copious literature on the proteins/ enzymes found in mRNA vaccines using the BLASTP approach (Appendix 1) 3. The reference literature, apart from being written in English, is also worded in an extremely technical style 4. They are given a specific functional sequence, aimed at the incorporation and expression of vaccine mRNA into the genome of the mitochondria (mtDNA) 5. The vaccine mRNA does not refer to the segment relating to the Spike protein, but to the entire viral genome 6. Therefore, the Ribosomes are not the final destination of the vaccine mRNA; instead, the Mitochondria are actually the seat of a DNA, accommodating the SARS-Cov2 genome in the same way as it does in the nuclear genome. 7. Both the Mitochondria and Ribosomes are located in the Ergastoplasm and are separated from the cytosol by an oxidation/reduction barrier 'in mV' (= one thousand times) three times higher than that of the cytosol. 8. Article from Science: Oxidized Redox State of Glutothiane in the Endoplasmic reticulum - 11 September 1992 volume 257, page 1496:15026 9. Thus only an mRNA that exits from both the nuclear and mitochondrial genomes has the enzyme support that allows it to reach the ribosomes, overcoming the resistance of the oxidation/reduction barrier 10. If this were not the case, RNA viruses would not lengthen the pathway, integrating before the genome and then reaching the ribosomes again as mRNA (it must be said that, in an RNA virus, the genomic RNA is the same as the RNA coming out of the nucleus), but would immediately aim to reach the ribosomes, as RNA vaccines are said to do. 11. Nano technologies are said to be the architects of this miraculous and rapid rush to the ribosomes of vaccine mRNA 12. These nano-technologies are nothing more than a sequence of proteins/enzymes - undeclared - which are now described here in their function in the main patterns 13. a) ryanodine receptorby managing the Ca++ ion flux channels, it first depolarises the mitochondrial membrane, which is the first step for mRNA entry and then it lowers the oxidative/reductive level of the ergastoplasm, activating an osmotic "Δ" (= delta-difference) around the mitochondria, releasing the protonated ions (=H + ) present in the virtual space of the two sheets forming the mitochondrial membrane into the area. 14. b) ABC transporter - ATP - binding protein By binding to ATP, it allows vaccine RNA to enter the mitochondria 15. c) Epical complex lysine methyltran spherase By providing mobility for the entering mRNA, it allows to reach the stretch of mitochondrial DNA most suitable for integration 16. d) DDLS - Typ - integrase/transpasase It simultaneously does the work performed before by reverse transcriptase and integrase, and then inserts a viral genome RNA, similar to the genomic RNA in RNA viruses, into the mitochondrial genome. 17. e) Pullulanase The hydrolysis of the (1 → 6) X - D glucoside bonds of mitochondrial DNA paves the way for the action of the

I6mA-Caps: a CapsuleNet-based framework for identifying DNA N6-methyladenine sites.

DNA N6-methyladenine (6mA) has been demonstrated to have an essential function in epigenetic modification in eukaryotic species in recent research. 6mA has been linked to various biological processes. It's critical to create a new algorithm that can rapidly and reliably detect 6mA sites in genomes to investigate their biological roles. The identification of 6mA marks in the genome is the first and most important step in understanding the underlying molecular processes, as well as their regulatory functions. In this article, we proposed a novel computational tool called i6mA-Caps which CapsuleNet based a framework for identifying the DNA N6-methyladenine sites. The proposed framework uses a single encoding scheme for numerical representation of the DNA sequence. The numerical data is then used by the set of convolution layers to extract low-level features. These features are then used by the capsule network to extract intermediate-level and later high-level features to classify the 6mA sites. The proposed network is evaluated on three datasets belonging to three genomes which are Rosaceae, Rice and Arabidopsis thaliana. Proposed method has attained an accuracy of 96.71%, 94% and 86.83% for independent Rosaceae dataset, Rice dataset and A.thaliana dataset respectively. The proposed framework has exhibited improved results when compared with the existing top-of-the-line methods. A user-friendly web-server is made available for the biological experts which can be accessed at: http://nsclbio.jbnu.ac.kr/tools/i6mA-Caps/. Supplementary data are available at Bioinformatics online.

P-025 Freezing of the semen sample by the patient at home is clinically effective

Abstract Study question To evaluate the efficacy of the method of freezing the seminal sample at home by the patient himself. Summary answer The freezing of the seminal sample at home is an effective method that brings the clinic closer to the patient, eliminating travel and stress. What is known already Seminal sample freezing for reproductive purpose is indicated in different situations: preservation of fertility, before a vasectomy, in patients with dysfunctions and problems obtaining a sample, and for those patients who cannot be present at the time of the oocyte retrieval. In many of these cases, the patient is under pressure to obtain the sample, in a clinical environment and at an exact time. Many experience this as a stressful situation, which in some cases forces them to travel if they live far from the clinic. Study design, size, duration In 2017, the home freezing protocol was validated with volunteers who obtained a semen sample at the clinic. After verifying that the quality of the sample was the same as if it were frozen by an expert biologist, we began offering this technique to our patients in 2019. We compared the results of the DO cycles with a control group to determine the efficacy of the protocol. Participants/materials, setting, methods Until December 2021, a total of 18 patients have used the Freezing Kit at home, 14 were residents in other EU countries, and 4 residents in Spain, but far from the clinic. As a control group, we used 608 oocyte donation cycles with a frozen sample. Patients who decided to freeze their sample at home receive a kit consisting of a travel tank with nitrogen vapors, a box with all the necessary material, and instructions. Main results and the role of chance Regarding the results of the 18 patients who have used the kit to freeze samples for their oocyte donation cycles, they have a mean of 44±7.1 years, and the samples had a post-thaw concentration of 48±30.5 M /ml with a mean mobility of 21.18±14.6%. No statistically significant differences were found in the fertilization rate compared to the control group (76.4±15.1% vs 77.8±18.3%; p = 0.485), blastocyst rate (61%±10 .5% vs 56.1±16.4%; p = 0.387) nor clinical pregnancy rate (62.8% vs 47.6%; p = 0.251). The fact of having a live birth from a sample frozen by the patient himself has just demonstrated the safety and usefulness of this novel freezing protocol. Limitations, reasons for caution The number of patients who have undergone this process is limited due to the nature of the treatment. Wider implications of the findings These results demonstrate the clinical utility of this freezing method for certain patients, where travel to the clinic may be inconvenient. Demonstrating that it is an effective method of cryopreservation of the seminal sample that brings the clinic closer to the patient, eliminating trips and stress. Trial registration number not applicable