Criteria for the selection, evaluation and application of traditional knowledge in contemporary health practice, education, research and policy: A systematic review.

BACKGROUND: Traditional and complementary medicine (T&CM) is highly utilised and draws on traditional knowledge (TK) as evidence, raising a need to explore how TK is currently used. OBJECTIVES: Examine criteria used to select, evaluate and apply TK in contemporary health contexts. METHODS: Systematic search utilising academic databases (AMED, CINAHL, MEDLINE, EMBASE, SSCI, ProQuest Dissertations Theses Global), Trip clinical database and Google search engine. Citations and reference lists of included articles were searched. Reported use of TK in contemporary settings was mapped against a modified 'Exploration-Preparation-Implementation-Sustainment' (EPIS) implementation framework. RESULTS: From the 54 included articles, EPIS mapping found TK is primarily used in the Exploration phase of implementation (n = 54), with little reporting on Preparation (n = 16), Implementation process (n = 6) or Sustainment (n = 4) of TK implementation. Criteria used in selection, evaluation and application of TK commonly involved validation with other scientific/traditional evidence sources, or assessment of factors influencing knowledge translation. DISCUSSION: One of the difficulties in validation of TK (as a co-opted treatment) against other evidence sources is comparing like with like as TK often takes a holistic approach. This complicates further planning and evaluation of implementation. CONCLUSION: This review identifies important criteria for evaluating current and potential contemporary use of TK, identifying gaps in research and practice for finding, appraising and applying relevant TK studies for clinical care.

Exploring Criteria for the Translation of Traditional Knowledge Within Contemporary Clinical Practice, Research, Policy, and Education: A Stakeholder Forum.

Background: The 2018 Declaration of Astana identifies traditional knowledge (TK) as one of the drivers for strengthening primary health care systems through the use of technology (traditional medicines) and knowledge and capacity building (traditional practitioners). While TK underpins both traditional practice and the use of traditional medicines, facilitating the use of TK in contemporary health care systems has been difficult to achieve. The aim of this study was to identify key factors related to the translation of TK into contemporary settings to help establish tools to support the knowledge translation process. Methods: This study used World Café methodology to collect the observations, ideas, and perspectives of experts who use TK in their practice. These experts (n = 9) were from a variety of contexts, including clinical practice, research, education, policy, and consumer advocacy, participated in the 1-day event. Data were collected into NVivo 12 software and analyzed using inductive-deductive thematic analysis. Results: Thematic analysis identified five themes: the need to define the elements required for critical evaluation of sources of TK as evidence, the importance of applying a tradition-centric lens when translating TK for contemporary use, the need to bridge gaps between TK and its contemporary applications, the value of critically evaluating the TK translation process itself, and the recognition of traditions as living systems. Taken together, the themes showed holistic interpretation of the translation process that incorporates critical analysis of the TK itself and accountable, transparent, and ethical processes of translation that consider safety, socioeconomical and intellectual property impacts of TK in contemporary use. Conclusions: Stakeholders identified TK as a valid and important source of evidence that should guide practice in a range of contemporary settings (e.g., policy and clinical practice), and outlined important consideration for critiquing, evaluating, communicating, and using TK within these settings.

Providing naturopathic care in the community: Perspectives of recent naturopathy graduates in Australia.

BACKGROUND AND PURPOSE: Naturopathy is a traditional medicine system originating from Europe and naturopathic practitioners provide care to 6.2% of Australians in a 12-month period. Australian naturopathic programs have undertaken a slow transition over the last 20 years from Advanced diploma to Bachelor degree as the minimum level of qualification for entry into the profession. This study aimed to understand and describe the experience of naturopathic graduates completing Bachelor degree and transitioning to provide naturopathic care in the community. MATERIALS AND METHODS: Qualitative semi-structured phone interviews were conducted with graduates of Bachelor's degree naturopathy programs, within five years of completing their studies. The data were analysed using Framework analysis methods. RESULTS: The analysis identified three related themes: (1) Love for looking after patients, but clinical practice is not easy, (2) Finding a place in the naturopathic profession and in the health system; and (3) Protecting the future of the profession and practice through registration. CONCLUSION: Graduates of Australian Bachelor's degree naturopathic programs face challenges as they attempt to find a place within their professional community. By identifying these challenges the profession's leaders may be able to develop initiatives to better support graduates and increase the success of new naturopaths.

The effects of kefir consumption on human health: a systematic review of randomized controlled trials.

CONTEXT: Kefir, a traditional, fermented-milk beverage, has increasingly been promoted for various health benefits. The evidence from systematic reviews, however, is limited. OBJECTIVE: Evidence from randomized controlled trials testing oral consumption of fermented-milk kefir on any outcome of human health or disease. DATA SOURCES: A systematic search of 4 electronic databases (PubMed, Scopus, Allied and Complementary Medicine Database, and Cochrane Trials) from inception to July 31, 2021, was conducted. DATA EXTRACTION: Data extraction and risk-of-bias assessments were conducted by 2 reviewers independently. DATA ANALYSIS: A total of 18 publications reporting the results of 16 studies were included. Per the narrative analysis, fermented-milk kefir may have potential as a complementary therapy in reducing oral Streptococcus mutans, thereby reducing dental caries risk, and in Helicobacter pylori eradication therapy. Kefir may further aid treatment of adult dyslipidemia and hypertension, although evidence was very limited. Safety was only assessed in 5 of the 18 included publications, and 12 of the studies had an overall high risk for bias. CONCLUSION: Kefir is a dairy product with a unique microbiological profile that appears to be a safe for generally healthy populations to consume. However, efficacy and safety data from high-quality human trials are essential before any recommendations may be made for conditions of the oral and gastric microbiota and metabolic health. SYSTEMATIC REVIEW REGISTRATION: PROSPERO registration no. CRD42020211494.

Ayurvedic formulations: Potential COVID-19 therapeutics?

BACKGROUND: While Molnupiravir and Paxlovid have recently been approved for use in some countries, there are no widely available treatments for COVID-19, the disease caused by SARS-CoV-2 infection. Herbal extracts have been used to treat respiratory clinical indications by Ayurvedic medicine practitioners with minimal adverse reactions and intense research efforts are currently under way to develop some of these formulations for COVID-19 treatment. METHODS: Literature search for in silico, in vitro, in vivo, and clinical studies on the topic of Ayurvedic formulations for potential COVID-19 treatment, in order to present the current state of current knowledge by integrating information across all systems. RESULTS: The search yielded 20 peer reviewed articles on in silico studies examining the interaction of phytoconstituents of popular Ayurvedic formulations with SARS-CoV-2 components and its receptors; five articles on preclinical investigations of the ability of selected Ayurvedic formulations to inhibit functions of SARS-CoV-2 proteins; and 51 completed clinical trials on the efficacy of using Ayurvedic formulations for treatment of mild to moderate COVID-19. Clinical data was available from 17 of the 51 trials. There was a considerable overlap between formulations used in the in silico studies and the clinical trials. This finding was unexpected as there is no clearly stated alignment between studies and the traditional pathway to drug discovery- basic discovery leading to in vitro and in vivo proof of concept, followed by validation in clinical trials. This was further demonstrated in the majority of the in silico studies where focus was on potential antiviral mechanisms, while the clinical trials were focused on patient recovery using oral treatments. In all 17 clinical trials where data was available, Ayurvedic treatments lead to a shorter period to recovery in participants with COVID-19. CONCLUSION: The most commonly used Ayurvedic treatments for management of respiratory symptoms associated with SARS-CoV-2 infection appear to have prophylactic and/or therapeutic properties. It would be of particular interest to assess synergistic and concomitant systemic effects and antiviral activities of individual phytoconstituents and their combinations in the Ayurvedic treatments.

The effects of Hedera helix on viral respiratory infections in humans: A rapid review.

BRIEF OVERVIEW: Based on the evidence identified in this rapid review, Hedera helix preparations and herbal complex preparations including H. helix may be a therapeutic option for treating early symptoms of respiratory tract infections. The best effectiveness for H. helix preparations has been proven for coughing, as an expectorant and to reduce the frequency and intensity of cough. Only weak evidence was found for all other researched symptoms. Both adults and children tolerate H. helix well. Currently, there is insufficient evidence to recommend the use of this supplement in the treatment or prevention of COVID-19. However, the current evidence justifies further research to better understand its applicability in coronavirus infections. VERDIC: tCurrent evidence suggests H. helix may improve the frequency and intensity of cough associated with viral respiratory infection. The overall applicability of additional findings is limited by the poorly defined outcome measures employed. However, studies focused explicitly on expectoration did report an increased conversion from dry to productive cough, and an improvement in expectoration amount, consistency and colour. These effects may be explained by a related finding of reduced oropharyngeal congestion and improved inflammatory markers (erythrocyte sedimentation rate and c-reactive protein). A decrease in frequency of night cough and respiratory pain was also reported, as was improved sleep quality and reduced cough-related sleep disturbance.Some studies also measured general respiratory tract infection symptoms and identified clinical improvement or resolution of fever, fatigue, sore throat, sneezing, wheezing, nasal congestion, post-nasal drip and body-ache. A reduced need for antibiotic prescriptions was also identified. While not consistently reported, the majority of studies also found H. helix reduced the overall severity of viral bronchitis and related conditions. Tolerability was rated as between 'good' and 'high'. Adverse events were rare or non-existent in almost all studies, and those that were reported were defined as non-serious and not drug-related.

Collaborative Active Learning Activities Promote Deep Learning in a Chemistry-Biochemistry Course.

Currently in higher education, there is a move towards providing more student-centred learning experiences, where students are actively involved in the learning process. To promote learner engagement and communication between peers, many educators utilise collaborative active learning activities. This study aimed to demonstrate that an active learning curriculum developed for a Chemistry-Biochemistry unit, allowed students to gain a deep understanding of the content, while developing key academic skills. In each face-to-face session of the Chemistry-Biochemistry unit, students participated in collaborative active learning activities including Participation+ and a variety of Padlet activities. The students were also challenged to develop their written communication skills, by taking part in a formative In-Class Writing Task. Survey results indicated that the active learning curriculum provided an engaging, interactive environment that was conducive to the students developing an understanding of the course's underlying concepts and developing key academic skills. The students communicated their deep understanding of the content verbally during active learning activities and in writing during the In-Class Writing Task, written assignment and final exam. Students who consistently communicated deep knowledge of the content during the In-Class Writing Task achieved high marks on the summative written assignment, final exam and unit total. This study clearly demonstrates that the active learning curriculum employed in the Chemistry-Biochemistry unit provided a collaborative and engaging learning environment, where many students developed a deep understanding of the content and acquired the skills to communicate their knowledge both orally and through written communication.

Classification of the human phox homology (PX) domains based on their phosphoinositide binding specificities.

Phox homology (PX) domains are membrane interacting domains that bind to phosphatidylinositol phospholipids or phosphoinositides, markers of organelle identity in the endocytic system. Although many PX domains bind the canonical endosome-enriched lipid PtdIns3P, others interact with alternative phosphoinositides, and a precise understanding of how these specificities arise has remained elusive. Here we systematically screen all human PX domains for their phospholipid preferences using liposome binding assays, biolayer interferometry and isothermal titration calorimetry. These analyses define four distinct classes of human PX domains that either bind specifically to PtdIns3P, non-specifically to various di- and tri-phosphorylated phosphoinositides, bind both PtdIns3P and other phosphoinositides, or associate with none of the lipids tested. A comprehensive evaluation of PX domain structures reveals two distinct binding sites that explain these specificities, providing a basis for defining and predicting the functional membrane interactions of the entire PX domain protein family.

Retromer's Role in Endosomal Trafficking and Impaired Function in Neurodegenerative Diseases.

The retromer complex is a highly conserved membrane trafficking assembly composed of three proteins - Vps26, Vps29 and Vps35 - that were identified over a decade ago in Saccharomyces cerevisiae (S. cerevisiae). Initially, mammalian retromer was shown to sort transmembrane proteins from the endosome to the trans-Golgi network (TGN), though recent work has identified a critical role for retromer in multiple trafficking pathways, including recycling to the plasma membrane and regulation of cell polarity. In recent years, genetic, cellular, pharmacological and animal model studies have identified retromer and its interacting proteins as being linked to familial forms of neurodegenerative diseases such as Alzheimer's (AD) and Parkinson's (PD). Here, this commentary will summarize recently identified point mutations in retromer linked to PD, and explore the molecular functions of retromer that may be relevant to disease progression.

Parkinson Disease-linked Vps35 R524W Mutation Impairs the Endosomal Association of Retromer and Induces α-Synuclein Aggregation.

Endosomal sorting is a highly orchestrated cellular process. Retromer is a heterotrimeric complex that associates with endosomal membranes and facilitates the retrograde sorting of multiple receptors, including the cation-independent mannose 6-phosphate receptor for lysosomal enzymes. The cycling of retromer on and off the endosomal membrane is regulated by a network of retromer-interacting proteins. Here, we find that Parkinson disease-associated Vps35 variant, R524W, but not P316S, is a loss-of-function mutation as marked by a reduced association with this regulatory network and dysregulation of endosomal receptor sorting. Expression of Vps35 R524W-containing retromer results in the accumulation of intracellular α-synuclein-positive aggregates, a hallmark of Parkinson disease. Overall, the Vps35 R524W-containing retromer has a decreased endosomal association, which can be partially rescued by R55, a small molecule previously shown to stabilize the retromer complex, supporting the potential for future targeting of the retromer complex in the treatment of Parkinson disease.

Functional characterization of retromer in GLUT4 storage vesicle formation and adipocyte differentiation.

Insulin-stimulated translocation of glucose transporter 4 (GLUT4) storage vesicles (GSVs), the specialized intracellular compartments within mature adipocytes, to the plasma membrane (PM) is a fundamental cellular process for maintaining glucose homeostasis. Using 2 independent adipocyte cell line models, human primary Simpson-Golabi-Behmel syndrome and mouse 3T3-L1 fibroblast cell lines, we demonstrate that the endosome-associated protein-sorting complex retromer colocalizes with GLUT4 on the GSVs by confocal microscopy in mature adipocytes. By use of both confocal microscopy and differential ultracentrifugation techniques, retromer is redistributed to the PM of mature adipocytes upon insulin stimulation. Furthermore, stable knockdown of the retromer subunit-vacuolar protein-sorting 35, or the retromer-associated protein sorting nexin 27, by lentivirus-delivered small hairpin RNA impaired the adipogenesis process when compared to nonsilence control. The knockdown of retromer decreased peroxisome proliferator activated receptor γ expression during differentiation, generating adipocytes with decreased levels of GSVs, lipid droplet accumulation, and insulin-stimulated glucose uptake. In conclusion, our study demonstrates a role for retromer in the GSV formation and adipogenesis.

Vps26B-retromer negatively regulates plasma membrane resensitization of PAR-2.

Retromer is a trimeric complex composed of Vps26, Vps29, and Vps35 and has been shown to be involved in trafficking and sorting of transmembrane proteins within the endosome. The Vps26 paralog, Vps26B, defines a distinct retromer complex (Vps26B-retromer) in vivo and in vitro. Although endosomally associated, Vps26B-retromer does not bind the established retromer transmembrane cargo protein, cation-independent mannose 6-phosphate receptor (CI-M6PR), indicating it has a distinct role to retromer containing the Vps26A paralog. In the present study we use the previously established Vps26B-expressing HEK293 cell model to address the role of Vps26B-retromer in trafficking of the protease activated G-protein coupled receptor PAR-2 to the plasma membrane. In these cells there is no apparent defect in the initial activation of the receptor, as evidenced by release of intracellular calcium, ERK1/2 signaling and endocytosis of activated receptor PAR-2 into degradative organelles. However, we observe a significant delay in plasma membrane repopulation of the protease activated G protein-coupled receptor PAR-2 following stimulation, resulting in a defect in PAR-2 activation after resensitization. Here we propose that PAR-2 plasma membrane repopulation is regulated by Vps26B-retromer, describing a potential novel role for this complex.

Soluble NSF attachment protein receptor molecular mimicry by a Legionella pneumophila Dot/Icm effector.

Upon infection, Legionella pneumophila uses the Dot/Icm type IV secretion system to translocate effector proteins from the Legionella-containing vacuole (LCV) into the host cell cytoplasm. The effectors target a wide array of host cellular processes that aid LCV biogenesis, including the manipulation of membrane trafficking. In this study, we used a hidden Markov model screen to identify two novel, non-eukaryotic soluble NSF attachment protein receptor (SNARE) homologs: the bacterial Legionella SNARE effector A (LseA) and viral SNARE homolog A proteins. We characterized LseA as a Dot/Icm effector of L. pneumophila, which has close homology to the Qc-SNARE subfamily. The lseA gene was present in multiple sequenced L. pneumophila strains including Corby and was well distributed among L. pneumophila clinical and environmental isolates. Employing a variety of biochemical, cell biological and microbiological techniques, we found that farnesylated LseA localized to membranes associated with the Golgi complex in mammalian cells and LseA interacted with a subset of Qa-, Qb- and R-SNAREs in host cells. Our results suggested that LseA acts as a SNARE protein and has the potential to regulate or mediate membrane fusion events in Golgi-associated pathways.

Structural basis for different phosphoinositide specificities of the PX domains of sorting nexins regulating G-protein signaling.

Sorting nexins (SNXs) or phox homology (PX) domain containing proteins are central regulators of cell trafficking and signaling. A subfamily of PX domain proteins possesses two unique PX-associated domains, as well as a regulator of G protein-coupled receptor signaling (RGS) domain that attenuates Gαs-coupled G protein-coupled receptor signaling. Here we delineate the structural organization of these RGS-PX proteins, revealing a protein family with a modular architecture that is conserved in all eukaryotes. The one exception to this is mammalian SNX19, which lacks the typical RGS structure but preserves all other domains. The PX domain is a sensor of membrane phosphoinositide lipids and we find that specific sequence alterations in the PX domains of the mammalian RGS-PX proteins, SNX13, SNX14, SNX19, and SNX25, confer differential phosphoinositide binding preferences. Although SNX13 and SNX19 PX domains bind the early endosomal lipid phosphatidylinositol 3-phosphate, SNX14 shows no membrane binding at all. Crystal structures of the SNX19 and SNX14 PX domains reveal key differences, with alterations in SNX14 leading to closure of the binding pocket to prevent phosphoinositide association. Our findings suggest a role for alternative membrane interactions in spatial control of RGS-PX proteins in cell signaling and trafficking.

The Vps35 D620N mutation linked to Parkinson's disease disrupts the cargo sorting function of retromer.

The retromer is a trimeric cargo-recognition protein complex composed of Vps26, Vps29 and Vps35 associated with protein trafficking within endosomes. Recently, a pathogenic point mutation within the Vps35 subunit (D620N) was linked to the manifestation of Parkinson's disease (PD). Here, we investigated details underlying the molecular mechanism by which the D620N mutation in Vps35 modulates retromer function, including examination of retromer's subcellular localization and its capacity to sort cargo. We show that expression of the PD-linked Vps35 D620N mutant redistributes retromer-positive endosomes to a perinuclear subcellular localization and that these endosomes are enlarged in both model cell lines and fibroblasts isolated from a PD patient. Vps35 D620N is correctly folded and binds Vps29 and Vps26A with the same affinity as wild-type Vps35. While PD-linked point mutant Vps35 D620N interacts with the cation-independent mannose-6-phosphate receptor (CI-M6PR), a known retromer cargo, we find that its expression disrupts the trafficking of cathepsin D, a CI-M6PR ligand and protease responsible for degradation of α-synuclein, a causative agent of PD. In summary, we find that the expression of Vps35 D620N leads to endosomal alterations and trafficking defects that may partly explain its action in PD.

A set of vertically integrated inquiry-based practical curricula that develop scientific thinking skills for large cohorts of undergraduate students.

Science graduates require critical thinking skills to deal with the complex problems they will face in their 21st century workplaces. Inquiry-based curricula can provide students with the opportunities to develop such critical thinking skills; however, evidence suggests that an inappropriate level of autonomy provided to underprepared students may not only be daunting to students but also detrimental to their learning. After a major review of the Bachelor of Science, we developed, implemented, and evaluated a series of three vertically integrated courses with inquiry-style laboratory practicals for early-stage undergraduate students in biomedical science. These practical curricula were designed so that students would work with increasing autonomy and ownership of their research projects to develop increasingly advanced scientific thinking and communication skills. Students undertaking the first iteration of these three vertically integrated courses reported learning gains in course content as well as skills in scientific writing, hypothesis construction, experimental design, data analysis, and interpreting results. Students also demonstrated increasing skills in both hypothesis formulation and communication of findings as a result of participating in the inquiry-based curricula and completing the associated practical assessment tasks. Here, we report the specific aspects of the curricula that students reported as having the greatest impact on their learning and the particular elements of hypothesis formulation and communication of findings that were more challenging for students to master. These findings provide important implications for science educators concerned with designing curricula to promote scientific thinking and communication skills alongside content acquisition.

Structural basis for endosomal trafficking of diverse transmembrane cargos by PX-FERM proteins.

Transit of proteins through the endosomal organelle following endocytosis is critical for regulating the homeostasis of cell-surface proteins and controlling signal transduction pathways. However, the mechanisms that control these membrane-transport processes are poorly understood. The Phox-homology (PX) domain-containing proteins sorting nexin (SNX) 17, SNX27, and SNX31 have emerged recently as key regulators of endosomal recycling and bind conserved Asn-Pro-Xaa-Tyr-sorting signals in transmembrane cargos via an atypical band, 4.1/ezrin/radixin/moesin (FERM) domain. Here we present the crystal structure of the SNX17 FERM domain bound to the sorting motif of the P-selectin adhesion protein, revealing both the architecture of the atypical FERM domain and the molecular basis for recognition of these essential sorting sequences. We further show that the PX-FERM proteins share a promiscuous ability to bind a wide array of putative cargo molecules, including receptor tyrosine kinases, and propose a model for their coordinated molecular interactions with membrane, cargo, and regulatory proteins.

Vps26A and Vps26B subunits define distinct retromer complexes.

The trimeric Vps29-Vps35-Vps26 sub-complex of retromer mediates retrograde transport of transmembrane proteins from endosomes to the trans-Golgi network. Our group has recently identified a Vps26 paralogue, Vps26B, which is able to suppress the expression of Vps26A when exogenously expressed in mammalian cells and defines a distinct retromer complex (Vps26B-retromer) in vivo and in vitro. In this study, we use HEK293 cells stably expressing either Vps26A-myc or Vps26B-myc to address the role of retromer cargo transport and subcellular localization of the two core retromer complexes as defined by the two mammalian-specific Vps26 paralogues. Vps26B-retromer, like Vps26A-retromer, associates with TBC1D5 and GOLPH3. In contrast, no interaction between Vps26B-retromer and cation-independent mannose 6-phosphate receptor (CI-M6PR) was detected, leading to a degradation of this receptor and an increase in cathepsin D secretion. Colocalization of Vps26 paralogues with different endosomally located Rab proteins shows prolonged association of Vps26B-retromer with maturing endosomes relative to Vps26A-retromer. Interestingly, the cycling of CI-M6PR is restored upon deletion of the variable Vps26B C-terminal region indicating that this region is directly responsible for the differential function of the two paralogues. In summary, we show that the two distinct retromer complexes defined by different Vps26 paralogues are not functionally equivalent and that the Vps26B C-terminal region can control cargo selection of the Vps26B-retromer.

Phox homology band 4.1/ezrin/radixin/moesin-like proteins function as molecular scaffolds that interact with cargo receptors and Ras GTPases.

Following endocytosis, the fates of receptors, channels, and other transmembrane proteins are decided via specific endosomal sorting pathways, including recycling to the cell surface for continued activity. Two distinct phox-homology (PX)-domain-containing proteins, sorting nexin (SNX) 17 and SNX27, are critical regulators of recycling from endosomes to the cell surface. In this study we demonstrate that SNX17, SNX27, and SNX31 all possess a novel 4.1/ezrin/radixin/moesin (FERM)-like domain. SNX17 has been shown to bind to Asn-Pro-Xaa-Tyr (NPxY) sequences in the cytoplasmic tails of cargo such as LDL receptors and the amyloid precursor protein, and we find that both SNX17 and SNX27 display similar affinities for NPxY sorting motifs, suggesting conserved functions in endosomal recycling. Furthermore, we show for the first time that all three proteins are able to bind the Ras GTPase through their FERM-like domains. These interactions place the PX-FERM-like proteins at a hub of endosomal sorting and signaling processes. Studies of the SNX17 PX domain coupled with cellular localization experiments reveal the mechanistic basis for endosomal localization of the PX-FERM-like proteins, and structures of SNX17 and SNX27 determined by small angle X-ray scattering show that they adopt non-self-assembling, modular structures in solution. In summary, this work defines a novel family of proteins that participate in a network of interactions that will impact on both endosomal protein trafficking and compartment specific Ras signaling cascades.

Characterisation of oxidized recombinant human galectin-1.

Oxidized human galectin-1 plays a role in the immune response to injured axons. Over-expression of galectin-1 by cancer, in combination with cancer associated oxidative stress suggests oxidized human galectin-1 may also play a role(s) in tumourigenesis. Here we generate milligram quantities of oxidized human galectin-1 and undertake biophysical characterization. The protein adopts a number of different states. Two separable oxidized forms are identified that exist as largely mono-disperse solutions at higher milligram/ml concentrations. The presence of disulphide bonds is confirmed for these two protein forms, as is their change in overall shape and loss of lectin activity. Our studies lead to production of a particular mono-disperse oxidized human galectin-1 species that is anticipated most optimal for investigations requiring milligram/ml concentrations such as X-ray crystallography.