Microbial signatures and continuum in endometrial cancer and benign patients.

BACKGROUND: Endometrial cancer is a multifactorial disease with inflammatory, metabolic and potentially microbial cues involved in disease pathogenesis. The endometrial cancer microbiome has been poorly characterised so far and studies have often overestimated bacterial biomass due to lack of integration of appropriate contamination controls. There is also a scarcity of evidence on the functionality of microbial microenvironments in endometrial cancer. This work addresses that knowledge gap by interrogating the genuine, contamination-free microbial signatures in the female genital tract and rectum of women with endometrial cancer and the mechanistic role of microbiome on carcinogenic processes. RESULTS: Here we sampled different regions of the reproductive tract (vagina, cervix, endometrium, fallopian tubes and ovaries) and rectum of 61 patients (37 endometrial cancer; 24 benign controls). We performed 16S rRNA gene sequencing of the V1-V2 hypervariable regions and qPCR of the 16S rRNA gene to qualitatively and quantitatively assess microbial communities and used 3D benign and endometrial cancer organoids to evaluate the effect of microbial products of L. crispatus, which was found depleted in endometrial cancer patients following primary analysis, on endometrial cell proliferation and inflammation. We found that the upper genital tract of a subset of women with and without endometrial cancer harbour microbiota quantitatively and compositionally distinguishable from background contaminants. Endometrial cancer was associated with reduced cervicovaginal and rectal bacterial load together with depletion of Lactobacillus species relative abundance, including L. crispatus, increased bacterial diversity and enrichment of Porphyromonas, Prevotella, Peptoniphilus and Anaerococcus in the lower genital tract and endometrium. Treatment of benign and malignant endometrial organoids with L. crispatus conditioned media exerted an anti-proliferative effect at high concentrations but had minimal impact on cytokine and chemokine profiles. CONCLUSIONS: Our findings provide evidence that the upper female reproductive tract of some women contains detectable levels of bacteria, the composition of which is associated with endometrial cancer. Whether this is a cause or consequence of cancer pathophysiology and what is the functional significance of this finding remain to be elucidated to guide future screening tools and microbiome-based therapeutics. Video Abstract.

3D printed rectal swabs for assessing the gut microbiome, metabolome and inflammation.

Investigating the gut microbiome and metabolome frequently requires faecal samples, which can be difficult to obtain. Previous studies have shown that rectal swabs are comparable to faecal samples for analysing gut microbiota composition and key metabolites. In this study, 3D printed rectal swabs were compared with conventional flocked swabs and faecal samples, due to the potential advantages 3D printing as a technique offers for swab production and development. 16S rRNA gene sequencing, qPCR and metabolite profiling (using 1H-NMR spectroscopy) were performed on swab and faecal samples from healthy participants. Faecal calprotectin and total protein analysis were performed on samples from inflammatory bowel disease (IBD) patients. There were no significant differences between both swab types and faecal samples when assessing key measures of alpha and beta diversity, and differences in the abundance of major phyla. There was a strong correlation between both swab types and faecal samples for all combined metabolites detected by NMR. In IBD patients, there was no significant difference in faecal calprotectin and total protein levels between both swab types and faecal samples. These data lead us to conclude that 3D printed swabs are equivalent to flocked swabs for the analysis of the gut microbiome, metabolome and inflammation.

'Something that helped the whole picture': Experiences of parents offered rapid prenatal exome sequencing in routine clinical care in the English National Health Service.

OBJECTIVES: In October 2020, rapid prenatal exome sequencing (pES) was introduced into routine National Health Service (NHS) care in England. This study aimed to explore parent experiences and their information and support needs from the perspective of parents offered pES and of health professionals involved in its delivery. METHODS: In this qualitative study, semi-structured interviews were conducted with 42 women and 6 male partners and 63 fetal medicine and genetic health professionals. Interviews were transcribed verbatim and analysed using thematic analysis. RESULTS: Overall views about pES were positive and parents were grateful to be offered the test. Highlighted benefits of pES included the value of the additional information for pregnancy management and planning for future pregnancies. An anxious wait for results was common, often associated with the need to make decisions near to 24 weeks in pregnancy when there are legal restrictions for late termination. Descriptions of dealing with uncertainty were also common, even when results had been returned. Many parents described pES results as informing decision-making around whether or not to terminate pregnancy. Some professionals were concerned that a non-informative result could be overly reassuring and highlighted that careful counselling was needed to ensure parents have a good understanding of what the result means for their pregnancy. Emotional support from professionals was valued; however, some parents felt that post-test support was lacking. CONCLUSION: Parents and professionals welcomed the introduction of pES. Results inform parents' decision-making around the termination of pregnancy. When there are no diagnostic findings or uncertain findings from pES, personalised counselling that considers scans and other tests are crucial. Directing parents to reliable online sources of information and providing emotional support throughout could improve their experiences of care.

Colonisation of hospital surfaces from low- and middle-income countries by extended spectrum ß-lactamase- and carbapenemase-producing bacteria.

Hospital surfaces can harbour bacterial pathogens, which may disseminate and cause nosocomial infections, contributing towards mortality in low- and middle-income countries (LMICs). During the BARNARDS study, hospital surfaces from neonatal wards were sampled to assess the degree of environmental surface and patient care equipment colonisation by Gram-negative bacteria (GNB) carrying antibiotic resistance genes (ARGs). Here, we perform PCR screening for extended-spectrum ß-lactamases (blaCTX-M-15) and carbapenemases (blaNDM, blaOXA-48-like and blaKPC), MALDI-TOF MS identification of GNB carrying ARGs, and further analysis by whole genome sequencing of bacterial isolates. We determine presence of consistently dominant clones and their relatedness to strains causing neonatal sepsis. Higher prevalence of carbapenemases is observed in Pakistan, Bangladesh, and Ethiopia, compared to other countries, and are mostly found in surfaces near the sink drain. Klebsiella pneumoniae, Enterobacter hormaechei, Acinetobacter baumannii, Serratia marcescens and Leclercia adecarboxylata are dominant; ST15 K. pneumoniae is identified from the same ward on multiple occasions suggesting clonal persistence within the same environment, and is found to be identical to isolates causing neonatal sepsis in Pakistan over similar time periods. Our data suggests persistence of dominant clones across multiple time points, highlighting the need for assessment of Infection Prevention and Control guidelines.

Identification of a Sub-Clinical Salmonella spp. Infection in a Dairy Cow Using a Commercially Available Stool Storage Kit.

Stool sampling is a useful tool for diagnosing gastrointestinal disease in veterinary medicine. The sub-clinical disease burden of Salmonella spp. in cattle can become significant for farmers. However, current methods of faecal sampling in a rural setting for diagnosis are not consistently sufficient for the preservation of Salmonella spp. in faeces. This study evaluated the use of a commercial stool storage kit for bacterial preservation in cow faecal samples compared to unpreserved stools placed into refrigeration at different time-points. A stool sample was collected per-rectum from one apparently healthy Holstein-Freisen cow. The sample was weighed and aliquoted into two sterile Falcon tubes and into two commercial kit tubes. The aliquots were then placed into refrigeration at 4 °C at 0, 24, and 96 h after processing. One commercial kit tube was not aliquoted and remained at ambient temperature. After 2 weeks, DNA was extracted from the samples and analysed using endpoint PCR, revealing a sub-clinical infection with Salmonella spp. The bacterium was best preserved when the stool was stored in the commercial kit at ambient temperature and re-homogenised immediately prior to DNA extraction. The unpreserved stool did not maintain obvious levels of Salmonella spp. after 24 h at ambient temperature. This commercial kit should be considered for use in the diagnosis of salmonellosis in cattle.

I AM FAMILY: Understanding the adapted sport experiences of family members of military personnel with physical and psychological illnesses and injuries through the lens of the Invictus Games.

Family are influential actors in adapted sport participation. However, little is known about their experiences with adapted sport. The current study sought to explore the experiences of families in adapted sport through the context of the Invictus Games, an international adapted sport competition for military personnel with physical and psychological illnesses and injuries that is unique in its inclusion of family programming. Family members (n = 21; partners, parents, siblings, and children) of Invictus Games Toronto 2017 competitors participated in semi-structured interviews. Data were analyzed using reflexive thematic analysis. Three themes were identified: recognition of family experiences during service and after onset of disability; creating a sense of belonging; and improving family knowledge and perceptions. This study provides insight regarding how adapted sport events can support the well-being of both individuals with illnesses and injuries and their families.

P22-A142†Validation of an improved closed system for dispensing serum eye drops.

PURPOSE: NHS Blood and Transplant Tissue and Eye Services provide a serum eye drop (SED) service to patients suffering from severe dry eye syndrome. Currently SED are dispensed using an automatic closed filling system (TF) manufactured by Meise Medizintechnik (Germany). An improved version (ATS) has recently been introduced by Meise, based on patient feedback on the TF system. ATS vials are easier to open, with a more secure, tamper evident closure and a better quality nozzle.To evaluate the suitability of ATS vials, a validation protocol, previously developed for TF vials, was repeated. It comprised assessment of their integrity following simulated storage and transport, and the stability and sterility of SED stored in them. METHOD: Firstly, a process simulation assessment was performed using bovine serum. Vials were filled, and frozen to -80oC. They were then removed from frozen storage and checked for damage, before being put into transport containers and shipped on a round-trip journey to simulate delivery to patients. On return the vials were thawed and the integrity of each vial checked visually and by application of a standard force.Subsequently a shelf-life study was carried out using three batches of human SED. The vials were initially frozen to -80oC, then stored for set time points of 1, 3, 6 and 12 months in a standard domestic freezer set at 20oC (to mimic a home freezer). At each time point, 10 vials were thawed and examined for integrity, and the sterility and stability of the contents. Stability was assessed by measuring serum albumin concentrations and sterility by testing for presence of microbial contamination, under aerobic and anaerobic conditions. RESULTS: No vial damage or leakage was found at any time point in the ATS vials. No microbial contamination was detected, and no change in albumin levels was detected in SED throughout the storage period. CONCLUSION: This study has demonstrated that the ATS vials are suitable for provision of SED for clinical use to patients. Feedback is now being gathered from a patient focus group relating to usability of the vials.

P10-A128†Trending of contamination rates across NHSBT eye banks.

INTRODUCTION: NHS Blood and Transplant Tissue and Eye Services (TES) is a human multi-tissue, tissue bank supplying tissue for transplant to surgeons throughout the UK. NHSBT has two Eye Banks.NHSBT investigated all our corneas discard due to contamination with the aim to review for any patterns. NHSBT Eye Banks performs initial Microbiology sampling on all Corneas in Corneas in Organ Culture Media at 7 Days. Corneas undergo a 2nd Microbiology sampling the day after the cornea is transferred into dextran median. MATERIALS AND METHODS: Any Microbiology positive media Identified pre-transplant are sent to NHSBT's Microbiology Reference Laboratory (MSL) for Identification. Any organisms which are identified post-dispatch are sent to a Referral Laboratory for rapid Identification and Sensitivity/Specificity Testing. FILTON EYE BANK: Contaminated Corneas in Organ Media: 2018- 28 (0.91%) 2019 -45 (1.10%), 2020- 27 (1.03%), 2021- 39 (1.41%), 2022- 43 (2.1%) (until 15/08/22)Most common Identified Organisms: C.Ablicans C. glabrata C.paraphilotisContaminated In Dextran Pre-Transplant: 2018- 4 (0.17%) 2019 -6 (0.18%), 2020- 9 (0.46%), 2021- 0 (0%), 2022- 3 (0.3%) (until 15/08/22). Most common Identified Organisms: Bacillus speciesContaminated in Dextran Post Transplant: 2018- 0 (0%) 2019 -8 (0.23%), 2020- 2(0.10%), 2021- 2 (0.08%), 2022- 1 (0.11%) (until 15/08/22). Most common Identified Organisms: Bacillus speciesDavid Lucas Eye Bank: Contaminated Corneas in Organ Media: 2020- 20(1.8%), 2021- 37(1.96%), 2022- 21(1.4%) (until 15/08/22). Most common Identified Organisms: C.Ablicans C. glabrata C.KefyrContaminated In Dextran Pre-Transplant: 2020- 6(0.8%), 2021- 2(0.14%), 2022- 1(0.08%) (until 15/08/22). Most common Identified Organisms: Bacillus speciesContaminated in Dextran Post Transplant: 2020- 2 (0.26%), 2021- 1 (0.07%), 2022- 2 (0.16%) (until 15/08/22). Most common Identified Organisms: Bacillus species DISCUSSION: Processes and facilities are of same standard between the two NHSBT Eye Banks and contamination rates are comparable. contamination is only identified in Approx1% of corneas processed. Corneas where growth is identified in Dextran is less than 1% of corneas Issued. Of the positive Microbiology samples identified post-Transplant, were mostly identified as Environmental Bacteria and had no patient impact on patient and assumed to have been contaminated by the operator.

A scoping review of military and Veteran families within international suicidality and suicide prevention research.

The impacts of suicidality on families are well known, which is particularly relevant in at-risk populations, such as active duty military personnel and Veteran communities. This scoping review describes how military and Veteran families have been conceptualized within suicide prevention research. A systematic, multi-database search was conducted, and 4,835 studies were screened. All included studies underwent quality assessment. Bibliographic, participant, methodological, and family-relevant data was extracted and descriptively analyzed into Factors, Actors, and Impacts. In total, 51 studies (2007 - 2021) were included. Most studies focused on suicidality rather than suicide prevention. Factor studies described family constructs as a suicidality risk or protective factor for military personnel or Veterans. Actor studies described families' roles or responsibilities to act in relation to the suicidality of military personnel or Veterans. Impacts studies described the impacts of suicidality on military and Veteran family members. The search was limited to English language studies. There were few studies on suicide prevention interventions for or including military and Veteran family members. Family was typically considered peripheral to the military personnel or Veteran experiencing suicidality. However, there was also emerging evidence of suicidality and its consequences in military-connected family members.

Gut microbiota fermentation profiles of pre-digested mycoprotein (Quorn) using faecal batch cultures in vitro: a preliminary study.

High-fibre diets are beneficial for many health outcomes via a wide range of mechanisms including gut microbiota fermentation-derived short-chain fatty acid (SCFAs) production. Mycoprotein (marketed as Quorn) is a food high in fibre (>6 g/100 g wet weight (ww)) and protein (13 g/100 g ww) which has been shown to have positive effects on glycemic control and appetite in humans. Nevertheless, the mechanisms underpinning this are poorly understood. Here, we investigate the changes in gut microbiota α- and ß-diversity, pH and SCFAs production in faecal batch cultures supplemented with pre-digested mycoprotein (Quorn), soy, chicken and control (unsupplemented) using eight fresh stools from healthy donors. The results showed that pre-digested mycoprotein did not alter pH (p = .896), α- or ß-diversity of the gut microbiota when compared to the control, soy, and chicken. Nevertheless, chicken led to a significant increase in total SCFAs post-24 h vs. control (+57.07 mmol/L, p = .01). In particular, propionate increased when compared to soy (+19.59 mmol/L, p = .03) and the control (+23.19 mmol/L, p < .01). No other differences in SCFAs were detected. In conclusion, pre-digested mycoprotein was not fermented in vitro by healthy gut microbiota in the settings of this experiment.

Pathobionts in the tumour microbiota predict survival following resection for colorectal cancer.

BACKGROUND AND AIMS: The gut microbiota is implicated in the pathogenesis of colorectal cancer (CRC). We aimed to map the CRC mucosal microbiota and metabolome and define the influence of the tumoral microbiota on oncological outcomes. METHODS: A multicentre, prospective observational study was conducted of CRC patients undergoing primary surgical resection in the UK (n = 74) and Czech Republic (n = 61). Analysis was performed using metataxonomics, ultra-performance liquid chromatography-mass spectrometry (UPLC-MS), targeted bacterial qPCR and tumour exome sequencing. Hierarchical clustering accounting for clinical and oncological covariates was performed to identify clusters of bacteria and metabolites linked to CRC. Cox proportional hazards regression was used to ascertain clusters associated with disease-free survival over median follow-up of 50 months. RESULTS: Thirteen mucosal microbiota clusters were identified, of which five were significantly different between tumour and paired normal mucosa. Cluster 7, containing the pathobionts Fusobacterium nucleatum and Granulicatella adiacens, was strongly associated with CRC (PFDR = 0.0002). Additionally, tumoral dominance of cluster 7 independently predicted favourable disease-free survival (adjusted p = 0.031). Cluster 1, containing Faecalibacterium prausnitzii and Ruminococcus gnavus, was negatively associated with cancer (PFDR = 0.0009), and abundance was independently predictive of worse disease-free survival (adjusted p = 0.0009). UPLC-MS analysis revealed two major metabolic (Met) clusters. Met 1, composed of medium chain (MCFA), long-chain (LCFA) and very long-chain (VLCFA) fatty acid species, ceramides and lysophospholipids, was negatively associated with CRC (PFDR = 2.61 × 10-11); Met 2, composed of phosphatidylcholine species, nucleosides and amino acids, was strongly associated with CRC (PFDR = 1.30 × 10-12), but metabolite clusters were not associated with disease-free survival (p = 0.358). An association was identified between Met 1 and DNA mismatch-repair deficiency (p = 0.005). FBXW7 mutations were only found in cancers predominant in microbiota cluster 7. CONCLUSIONS: Networks of pathobionts in the tumour mucosal niche are associated with tumour mutation and metabolic subtypes and predict favourable outcome following CRC resection. Video Abstract.

The gut microbiota and metabolome are associated with diminished COVID-19 vaccine-induced antibody responses in immunosuppressed inflammatory bowel disease patients.

BACKGROUND: Patients with inflammatory bowel disease (IBD) treated with anti-TNF therapy exhibit attenuated humoral immune responses to vaccination against SARS-CoV-2. The gut microbiota and its functional metabolic output, which are perturbed in IBD, play an important role in shaping host immune responses. We explored whether the gut microbiota and metabolome could explain variation in anti-SARS-CoV-2 vaccination responses in immunosuppressed IBD patients. METHODS: Faecal and serum samples were prospectively collected from infliximab-treated patients with IBD in the CLARITY-IBD study undergoing vaccination against SARS-CoV-2. Antibody responses were measured following two doses of either ChAdOx1 nCoV-19 or BNT162b2 vaccine. Patients were classified as having responses above or below the geometric mean of the wider CLARITY-IBD cohort. 16S rRNA gene amplicon sequencing, nuclear magnetic resonance (NMR) spectroscopy and bile acid profiling with ultra-high-performance liquid chromatography mass spectrometry (UHPLC-MS) were performed on faecal samples. Univariate, multivariable and correlation analyses were performed to determine gut microbial and metabolomic predictors of response to vaccination. FINDINGS: Forty-three infliximab-treated patients with IBD were recruited (30 Crohn's disease, 12 ulcerative colitis, 1 IBD-unclassified; 26 with concomitant thiopurine therapy). Eight patients had evidence of prior SARS-CoV-2 infection. Seventeen patients (39.5%) had a serological response below the geometric mean. Gut microbiota diversity was lower in below average responders (p = 0.037). Bilophila abundance was associated with better serological response, while Streptococcus was associated with poorer response. The faecal metabolome was distinct between above and below average responders (OPLS-DA R2X 0.25, R2Y 0.26, Q2 0.15; CV-ANOVA p = 0.038). Trimethylamine, isobutyrate and omega-muricholic acid were associated with better response, while succinate, phenylalanine, taurolithocholate and taurodeoxycholate were associated with poorer response. INTERPRETATION: Our data suggest that there is an association between the gut microbiota and variable serological response to vaccination against SARS-CoV-2 in immunocompromised patients. Microbial metabolites including trimethylamine may be important in mitigating anti-TNF-induced attenuation of the immune response. FUNDING: JLA is the recipient of an NIHR Academic Clinical Lectureship (CL-2019-21-502), funded by Imperial College London and The Joyce and Norman Freed Charitable Trust. BHM is the recipient of an NIHR Academic Clinical Lectureship (CL-2019-21-002). The Division of Digestive Diseases at Imperial College London receives financial and infrastructure support from the NIHR Imperial Biomedical Research Centre (BRC) based at Imperial College Healthcare NHS Trust and Imperial College London. Metabolomics studies were performed at the MRC-NIHR National Phenome Centre at Imperial College London; this work was supported by the Medical Research Council (MRC), the National Institute of Health Research (NIHR) (grant number MC_PC_12025) and infrastructure support was provided by the NIHR Imperial Biomedical Research Centre (BRC). The NIHR Exeter Clinical Research Facility is a partnership between the University of Exeter Medical School College of Medicine and Health, and Royal Devon and Exeter NHS Foundation Trust. This project is supported by the National Institute for Health Research (NIHR) Exeter Clinical Research Facility. The views expressed are those of the authors and not necessarily those of the NIHR or the UK Department of Health and Social Care.

Gut Microbiota Modulation of Efficacy and Toxicity of Cancer Chemotherapy and Immunotherapy.

Accumulating evidence supports not only the functional role of the gut microbiome in cancer development and progression but also its role in defining the efficacy and toxicity of chemotherapeutic agents (5-fluorouracil, cyclophosphamide, irinotecan, oxaliplatin, gemcitabine, methotrexate) and immunotherapeutic compounds (anti-programmed death-ligand 1/anti-programmed cell death protein 1 and anti-cytotoxic T-lymphocyte-associated antigen 4). This evidence is supported in numerous in vitro, animal, and clinical studies that highlight the importance of microbial mechanisms in defining therapeutic responses. The microbiome therefore shapes oncologic outcomes and is now being leveraged for the development of novel personalized therapeutic approaches in cancer treatment. However, if the microbiome is to be successfully translated into next-generation oncologic treatments, a new multimodal model of the oncomicrobiome must be conceptualized that incorporates gut microbial cometabolism of pharmacologic agents into cancer care. The objective of this review is therefore to outline the current knowledge of oncologic pharmacomicrobiomics and to describe how the multiparametric functions of the gut microbiome influence treatment response across cancer types. The secondary objective is to propose innovative approaches for modulating the gut microbiome in clinical environments that improve therapy efficacy and diminish toxic effects derived from antineoplastic agents for patient benefit.

Utility of head computed tomography for older adults with suspected delirium in the emergency department: A retrospective observational study.

BACKGROUND: Delirium in older people is associated with significant morbidity and mortality and has life-threatening etiologies making prompt recognition essential. Computed tomography of the head (CT-head) may have a role in determining the cause of delirium; however, inpatient studies suggest it is overused. There is a paucity of emergency department (ED)-based research surrounding the use of CT-head in delirium. This study aims to describe the utility of CT-head in older patients presenting to the ED with symptoms of delirium. METHODS: We conducted a retrospective chart review of patients 65 years and older with symptoms of delirium who visited local EDs over a 3.5-year period (2016-2020). We compared patients who did and did not receive CT-head. Our primary objective was to determine the proportion of acute findings in patients who received CT-head. Our secondary objectives were to describe the proportions of patients who did and did not receive CT-head in terms of their demographics, presenting symptoms, disposition, and indications for and results of CT-head scans. Chi-square tests were utilized for comparisons. RESULTS: A total of 630 encounters were identified through database searching; 526 met inclusion criteria. Thirty-four were excluded for presenting directly to consultants, leaving 492 included encounters. Of those who received a CT-head (n = 279), 13 (4.7%) had acute findings. Of the encounters with acute findings, four (30.77%) had focal neurological deficits (FND), and two (15.38%) had Glasgow Coma Scale (GCS) score < 14. Patients without CT-head (n = 213) were more likely to be discharged (p < 0.01) and less likely to have a FND (p < 0.01). CONCLUSIONS: CT-head is ordered for over half of older ED patients with symptoms of delirium despite infrequent acute findings. Acute findings typically occur in the context of symptoms suggestive of intracranial abnormalities such as FND or GCS < 14. This suggests physicians should be more selective when ordering CT-heads in patients with symptoms of delirium.

Kinematic Analysis of Sequential Partial-Midfoot Arthrodesis in Simulated Gait Cadaver Model.

BACKGROUND: Primary tarsometatarsal (TMT) arthrodesis is gaining popularity in the surgical treatment of Lisfranc injuries. However, few studies have evaluated biomechanical effects of TMT arthrodesis. The purpose of this study was to compare the kinematics of joints adjacent to the midfoot during simulations of stance before and after sequential arthrodesis of the first, second, and third TMT joints. METHODS: Ten midtibia cadaveric specimens were loaded on a 6-degree-of-freedom robotic gait simulator. Motion capture cameras were used to collect joint kinematics throughout simulations of the stance phase. Simulations were performed for the intact and sequential arthrodesis conditions of the first, second, and third TMT joints. The sagittal, coronal, and transverse plane rotational kinematics of the intact condition were compared to kinematics after each sequential arthrodesis condition. RESULTS: Sequential arthrodesis of the first and second TMT joints had no significant effect on ankle, subtalar, talonavicular, and first metatarsophalangeal joint motion during simulated stance when compared to the intact condition. In contrast, inclusion of the third TMT joint into the sequential arthrodesis significantly increased subtalar inversion (P = .032) in late stance and increased range of motion values in the ankle and subtalar joints by 2.1 degrees (P = .009) and 2.8 degrees (P = .014), respectively. CONCLUSION: Sequential primary arthrodesis induced changes to ankle and adjacent joint kinematics during stance phase simulations, although not until the third TMT joint was included into the primary arthrodesis. The significant changes to kinematics due to arthrodesis of the first, second, and third TMT joints were small. CLINICAL RELEVANCE: The minimal changes in sagittal, coronal, and transverse plane rotational kinematics support the positive clinical outcomes reported in the literature for primary partial arthrodesis of Lisfranc injuries. The inclusion of the third TMT joint should be done judiciously.