Neoantigen-targeted CD8+ T cell responses with PD-1 blockade therapy.

Neoantigens are peptides derived from non-synonymous mutations presented by human leukocyte antigens (HLAs), which are recognized by antitumour T cells1-14. The large HLA allele diversity and limiting clinical samples have restricted the study of the landscape of neoantigen-targeted T cell responses in patients over their treatment course. Here we applied recently developed technologies15-17 to capture neoantigen-specific T cells from blood and tumours from patients with metastatic melanoma with or without response to anti-programmed death receptor 1 (PD-1) immunotherapy. We generated personalized libraries of neoantigen-HLA capture reagents to single-cell isolate the T cells and clone their T cell receptors (neoTCRs). Multiple T cells with different neoTCR sequences (T cell clonotypes) recognized a limited number of mutations in samples from seven patients with long-lasting clinical responses. These neoTCR clonotypes were recurrently detected over time in the blood and tumour. Samples from four patients with no response to anti-PD-1 also demonstrated neoantigen-specific T cell responses in the blood and tumour to a restricted number of mutations with lower TCR polyclonality and were not recurrently detected in sequential samples. Reconstitution of the neoTCRs in donor T cells using non-viral CRISPR-Cas9 gene editing demonstrated specific recognition and cytotoxicity to patient-matched melanoma cell lines. Thus, effective anti-PD-1 immunotherapy is associated with the presence of polyclonal CD8+ T cells in the tumour and blood specific for a limited number of immunodominant mutations, which are recurrently recognized over time.

Predictors of complication after groin dissection: a single-centre experience.

BACKGROUND: Inguinal lymphadenectomy (ILND) has historically been associated with substantial morbidity. The objective of this study was to obtain contemporary ILND morbidity rates and to identify potentially preventable risk factors. METHODS: We carried out a retrospective review of medical records for all superficial, deep, and combination groin dissections performed at a single, high-volume academic centre between January 2007 and December 2020. We collected data points for patient, disease, and surgery characteristics, and cancer outcomes. The outcome of interest was any complication within 30 days of surgery. Complications included wound infection, wound necrosis or disruption, seroma, drainage procedure, hematoma, and lymphedema. We performed multivariate logistic regression using SAS version 9.4. RESULTS: We identified 139 patients having undergone 89 superficial, 12 deep, and 38 combined dissection types, respectively. Melanoma accounted for 84.9% of cases. Of these patients, 56.1% had an adverse postoperative event within 30 days. Increasing age (odds ratio [OR] 1.04, 95% confidence interval [CI] 1.01-1.07, p < 0.01) and number of positive lymph nodes harvested (OR 1.22, 95% CI 1.00-1.50, p = 0.05) were associated with more complications. Patients with deep dissection showed a lower likelihood of complications than those with superficial dissection (OR 0.15, 95% CI 0.03-0.84, p < 0.05). CONCLUSION: Complication rates after ILND remain high. We identified a number of risk factors, providing opportunities for better selection and prevention.

The Association between Prolonged Antibiotic Use in Culture Negative Infants and Length of Hospital Stay and Total Hospital Costs.

OBJECTIVE: This study aimed to determine if prolonged antibiotic use at birth in neonates with a negative blood culture increases the total cost of hospital stay. STUDY DESIGN: This was a retrospective study performed at a 60-bed level IV neonatal intensive care unit. Neonates born <30 weeks of gestation or <1,500 g between 2016 and 2018 who received antibiotics were included. A multivariate linear regression analysis was conducted to determine if clinical factors contributed to increased hospital cost or length of stay. RESULTS: In total, 190 patients met inclusion criteria with 94 infants in the prolonged antibiotic group and 96 in the control group. Prolonged antibiotic use was associated with an increase length of hospital stay of approximately 31.87 days, resulting in a $69,946 increase in total cost of hospitalization. CONCLUSION: Prolonged antibiotics in neonates with negative blood culture were associated with significantly longer hospital length of stay and increased total cost of hospitalization. KEY POINTS: · Prolonged antibiotic use at birth is associated with prolonged hospital stay.. · Prolonged antibiotic use at birth is associated with increased cost of hospitalization.. · Prolonged antibiotic use at birth is associated with increased days on total parenteral nutrition.. · Prolonged antibiotic use at birth is associated with increased subsequent courses of antibiotics..

Family economic pressure and early adolescents' prosocial behavior: The importance of considering the types of prosocial behavior.

BACKGROUND: Prior research findings are mixed regarding whether prosocial behavior is positively or negatively related to socioeconomic status and its correlates, such as economic pressure. This may be due to the lack of considerations for the type of prosocial behavior. AIMS: In this study, we aimed to examine how six types of prosocial behavior (i.e., public, anonymous, compliant, emotional, dire, and altruistic) are related to economic pressure among early adolescents. We hypothesized that family economic pressure would be associated with each type of prosocial behavior in differing ways. MATERIALS & METHODS: Participants were 11-14 years old (N = 143, Mage = 12.2 years, SDage = 0.87, 63 boys, 1 trans-identified boy, 55 girls), early adolescents and their parents. Among them, 54.6% were non-Hispanic/Latinx (NH/L) White, 23.8% were NH/L Black, 11.2% were NH/L Asian, 2.1% were NH/L Multiracial, and 8.4% were Hispanic/Latinx. Parents reported family economic pressure and adolescents' six types of prosocial behavior. RESULTS: Path analysis revealed that economic pressure was negatively associated with emotional and dire prosocial behavior over and above age, gender, and race/ethnicity. Family economic pressure was unrelated to public, anonymous, compliant, and altruistic prosocial behavior. DISCUSSION: These findings show some support for the Family Stress Model, such that economic stress might hinder youth's prosocial development. At the same time, youth may have similar levels of certain types of prosocial behavior regardless of their family's economic pressure. CONCLUSION: This research provided insight into the complex relation between economic pressure and youth's prosocial behavior which varies depending on the type of behavior.

Loss of CHCHD2 and CHCHD10 activates OMA1 peptidase to disrupt mitochondrial cristae phenocopying patient mutations.

Dominant mutations in the mitochondrial paralogs coiled-helix-coiled-helix (CHCHD) domain 2 (C2) and CHCHD10 (C10) were recently identified as causing Parkinson's disease and amyotrophic lateral sclerosis/frontotemporal dementia/myopathy, respectively. The mechanism by which they disrupt mitochondrial cristae, however, has been uncertain. Using the first C2/C10 double knockout (DKO) mice, we report that C10 pathogenesis and the normal function of C2/C10 are intimately linked. Similar to patients with C10 mutations, we found that C2/C10 DKO mice have disrupted mitochondrial cristae, because of cleavage of the mitochondrial-shaping protein long form of OPA1 (L-OPA1) by the stress-induced peptidase OMA1. OMA1 was found to be activated similarly in affected tissues of mutant C10 knock-in (KI) mice, demonstrating that L-OPA1 cleavage is a novel mechanism for cristae abnormalities because of both C10 mutation and C2/C10 loss. Using OMA1 activation as a functional assay, we found that C2 and C10 are partially functionally redundant, and some but not all disease-causing mutations have retained activity. Finally, C2/C10 DKO mice partially phenocopied mutant C10 KI mice with the development of cardiomyopathy and activation of the integrated mitochondrial integrated stress response in affected tissues, tying mutant C10 pathogenesis to C2/C10 function.

A prospective observational study evaluating the use of remote patient monitoring in ED discharged COVID-19 patients in NYC.

OBJECTIVES: We investigated whether continuous remote patient monitoring (RPM) could significantly reduce return Emergency Department (ED) revisits among coronavirus disease 2019 (COVID-19) patients discharged from the emergency Department. MATERIALS AND METHODS: A prospective observational study was conducted from a total of 2833 COVID-19 diagnosed patients who presented to the Montefiore Medical Center ED between September 2020-March 2021. Study patients were remotely monitored through a digital platform that was supervised 24/7 by licensed healthcare professionals. Age and time-period matched controls were randomly sampled through retrospective review. The primary outcome was ED revisit rates among the two groups. RESULTS: In our study, 150 patients enrolled in the RPM program and 150 controls were sampled for a total of 300 patients. Overall, 59.1% of the patients identified as Hispanic/Latino. The RPM group had higher body mass index (BMI) (29 (25-35) vs. 27 (25-31) p-value 0.020) and rates of hypertension (50.7% (76) vs. 35.8% (54) p-value 0.009). There were no statistically significant differences in rates of ED revisit between the RPM group (8% (12)) and control group (9.3% (14)) (OR: 0.863; 95% CI:0.413-1. 803; p- 0.695). DISCUSSION AND CONCLUSION: Our study explored the impact of continuous monitoring versus intermittent monitoring for reducing ED revisits in a largely underrepresented population of the Bronx. Our study demonstrated that continuous remote patient monitoring showed no significant difference in preventing ED revisits compared to non-standardized intermittent monitoring. However, potential other acute care settings where RPM may be useful for identifying high-risk patients for early interventions warrant further study.

CHCHD2 accumulates in distressed mitochondria and facilitates oligomerization of CHCHD10.

Mutations in paralogous mitochondrial proteins CHCHD2 and CHCHD10 cause autosomal dominant Parkinson Disease (PD) and Amyotrophic Lateral Sclerosis/Frontotemporal Dementia (ALS/FTD), respectively. Using newly generated CHCHD2, CHCHD10 and CHCHD2/10 double knockout cell lines, we find that the proteins are partially functionally redundant, similarly distributed throughout the mitochondrial cristae, and form heterodimers. Unexpectedly, we also find that CHCHD2/CHCHD10 heterodimerization increases in response to mitochondrial stress. This increase is driven by differences in the proteins' stability and mutual affinity: CHCHD2 is preferentially stabilized by loss of mitochondrial membrane potential, and CHCHD10 oligomerization depends on CHCHD2 expression. Exploiting the dependence of CHCHD10 oligomerization on CHCHD2, we developed a heterodimer incorporation assay and demonstrate that CHCHD2 and CHCHD10 with disease-causing mutations readily form heterodimers. As we also find that both proteins are highly expressed in human Substantia nigra and cortical pyramidal neurons, mutant CHCHD2 and CHCHD10 may directly interact with their wild-type paralogs in the context of PD and ALS/FTD pathogenesis. Together, these findings demonstrate that differences in the stability and mutual affinity of CHCHD2 and CHCHD10 regulate their heterodimerization in response to mitochondrial distress, revealing an unanticipated link between PD and ALS/FTD pathogenesis.

Epidemiology of Admissions From the Emergency Department Among Febrile Infants Younger Than 90 Days in the United States, 2002 to 2012.

BACKGROUND AND OBJECTIVES: Febrile infants younger than 90 days are often subjected to invasive diagnostic evaluation and hospitalization to monitor for serious bacterial infection (SBI) despite recent changes in SBI risk and advances in diagnostics. Nationwide provider practices, over time, are unknown for this group. Our objective was to determine the likelihood of admission and associated complications of care for such infants, particularly those at lowest risk for SBI. METHODS: This is a retrospective cohort study including multisite emergency department (ED) visits using administrative data from the Pediatric Emergency Care Applied Research Network Core Data Project. We examined uncomplicated febrile infants younger than 90 days seen in the ED from 2002 to 2012 who did not have an explicit infectious diagnosis, critical illness, or chronic condition. RESULTS: We identified 38,224 infants, among whom, 11,600 (31%) were admitted. We observed a 3% increase in ED admission every year during the study period (P < 0.05). Age less than 30 days and black race were independent predictors of admission (P < 0.01). We identified 227 complications of care in the ED. Among those with a specified cause, 49% were attributed to antibiotic use or procedures related to the diagnosis and empiric treatment of suspected infection. CONCLUSIONS: Despite the decreasing risk of SBI and diagnostic advances during the study period, the odds of hospital admission increased for febrile infants at low risk for sepsis. We also provide first data on therapeutic and diagnostic test-related adverse events in this setting. These data provide further justification for a consensus guideline on management of such infants.

Palifermin for prevention of oral mucositis in allogeneic hematopoietic stem cell transplantation: a single-institution retrospective evaluation.

PURPOSE: The purpose of this study is to assess the impact of palifermin on oral mucositis (OM) and its sequelae in patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) who were conditioned with fractionated total body irradiation (FTBI) and etoposide. METHODS: This retrospective chart review study compared the effect of palifermin on the development of OM in patients who received this agent during an allo-HSCT (n = 99) to those who did not (n = 30). The primary end points were severity and duration of OM. Secondary end points included requirements for opioids, total parenteral nutrition (TPN), and intensive oral care; incidence of infection; length of hospital stay; and overall survival. RESULTS: There was no significant difference in the incidence of all grades of OM, but incidence of severe OM was decreased in palifermin-exposed patients (34 vs 80 %, p < 0.0001). In patients who developed OM (all grades), the median duration of OM was shorter in palifermin-exposed patients (13 vs 18 days, p = 0.0001); there was no difference in the median duration of severe OM. Patients who received palifermin used less opioids and required a shorter duration of intensive oral care. There was no difference in duration of TPN, incidence of infection, length of hospital stay, and overall survival. CONCLUSIONS: Our findings demonstrated a significant benefit with the use of palifermin for allo-HSCT recipients who were conditioned with FTBI and etoposide. Palifermin can potentially improve quality of life for this patient population and reduce complications and resources used during the transplant process. A randomized clinical trial is required to confirm these results.

Calculated in situ tear oxygen tension under hybrid contact lenses.

OBJECTIVE: We calculated corneal surface oxygen tension under hybrid contact lenses (CLs) by extending existing models of CL oxygen diffusion to the circumstances of hybrid CL designs with tear vaults. METHODS: Theoretical oxygen tensions at the corneal surface, if tear mixing and exchange are excluded, are calculated for hybrid CL (modern, high oxygen-permeable rigid center) designs with a single chamber corneal model using a computer software spreadsheet. Several specific in vivo instances of hybrid CL tear vaults are measured by both slitlamp biomicroscopy and optical coherence tomography (OCT). RESULTS: No significant difference was found between central thickness of hybrid CL rigid portions measured by Vigor gauge and OCT. The amount of central tear vault thickness was found to be significantly greater, however, when interpolated from biomicroscope images (500-1,554 µm, simulating a poorly fit lens) than from either OCT images (0-91 µm) or direct measurement through OCT software (0-96 µm). Using all measurements, excluding any potential contributions from tear exchange or mixing, we predict corneal surface tear pO2 values under several hybrid CLs to range from zero under a lens with an excessive vault to 112 mm Hg under a lens with a minimal vault. CONCLUSIONS: Tear vaults beneath the gas permeable portion of the hybrid CLs of 100 µm or less resulted in acceptable corneal surface oxygen values. Where tear vaults are greater than about 100 µm, predicted anterior corneal surface pO2 values are less likely to be adequate for optimal corneal physiology. Any significant tear exchange, however, should increase these values. Although measuring tear thickness, we noted that the photographic biomicroscope and the OCT methods provided inconsistent results, and thus limited the reliability of our primary result.

Clinical Indications for Rapid Sequence MRI in Pediatric Neurosurgical Patients and the Limitations and Barriers to Implementation.

Rapid and fast magnetic resonance imaging (MRI) protocols have become increasingly popular for pediatric neurosurgical patients as they are a great way to reduce ionizing radiation and sedation. While their popularity has increased, there are hurdles to overcome when transitioning to using them clinically, such as cost, staffing training, and motion artifact. Through this paper, we developed a protocol for clinical applications where rapid MRI can be a substitute or adjuvant in diagnostic workup. Further, we outline the relevant literature for the use of RS-MRI for the spine, TBI, and hydrocephalus pathologies while expanding upon the limitations and logistical barriers when transitioning to their use, a few of which are discussed above. Through this, we conclude that RS-MRI can be used diagnostically for spinal pathologies such as syrinx and hydrocephalus. Further, its lack of sensitivity for TBI findings makes rapid sequence magnetic resonance imaging (RS-MRI) a strong adjuvant with other advanced imaging or computed tomography (CT) for traumatic brain injury (TBI) pathologies.

The relationship between smoking and periodontal disease. Review of literature and case report.

Cigarette smoking has been associated with tooth loss from periodontal disease for a long time. Smoking cessation has been shown to reverse these effects. The purpose of this paper is to review the current literature regarding the possible mechanisms for destruction of the periodontium caused by smoking and to present a protocol for the implementation of a smoking cessation program at New York University College of Dentistry.

Atypical Systemic Neonatal Herpes Diagnosed Incidentally in Twin Infants.

Neonatal herpes simplex virus (HSV) infections are serious infections that usually occur in the first few weeks of life. Infants generally present with mucocutaneous lesions, central nervous system infection, and/or systemic disease. In this case report, we describe a set of twins that had unexpected presentations of neonatal HSV. Twin A was diagnosed incidentally on routine eye exam, and Twin B was diagnosed only because his twin was found to be infected; both infants were still hospitalized and were beyond 1 month of age. These twins exhibited atypical manifestations that diverge from the 3 main categories of neonatal HSV and expand our understanding of the spectrum of disease.

Content Quality of YouTube Videos About Metastatic Breast Cancer in Young Women: Systematic Assessment.

BACKGROUND: Young women with metastatic breast cancer (MBC) are part of a digitally connected generation yet are underserved in terms of information needs. YouTube is widely used to find and identify health information. The accessibility of health-related content on social media together with the rare and marginalized experiences of young women with MBC and the digital media practices of younger generations imply a considerable likelihood that young women with MBC will seek information and community on the internet. OBJECTIVE: This study aims to assess the content quality of MBC YouTube videos and to identify themes in the experiences of young women with MBC based on YouTube videos. METHODS: A systematic assessment of MBC YouTube videos using the search term "metastatic breast cancer young" was conducted in August 2021. The search was performed in an incognito browser and with no associated YouTube or Google account. Search results were placed in order from most to least views. Title, date uploaded, length, poster identity, number of likes, and number of comments were collected. Understandability and actionability were assessed using the Patient Education Materials Assessment Tool (PEMAT); information reliability and quality were assessed with DISCERN. Themes, sponsorships, and health care professionals' and patients' narratives were reported. RESULTS: A total of 101 videos were identified. Of these, 78.2% (n=79) included sponsorships. The mean PEMAT scores were 78.8% (SD 15.3%) and 43.1% (SD 45.2%) for understandability and actionability, respectively. The mean DISCERN score was 2.44 (SD 0.7) out of 5. Identified themes included treatment (n=67, 66.3%), family relationship (n=46, 45.5%), and motherhood (n=38, 37.6%). CONCLUSIONS: YouTube videos about young women with MBC are highly understandable but demonstrate moderate rates of actionability, with low reliability and quality scores. Many have a commercial bias. While web-based materials have limitations, their potential to provide patient support is not fully developed. By acknowledging their patients' media habits, health care professionals can further develop a trusting bond with their patients, provide a space for open and honest discussions of web-based materials, and avoid any potential instances of confusion caused by misleading, inaccurate, or false web-based materials.

The Use of Web-Based Patient Reviews to Assess Medical Oncologists' Competency: Mixed Methods Sequential Explanatory Study.

BACKGROUND: Patients increasingly use web-based evaluation tools to assess their physicians, health care teams, and overall medical experience. OBJECTIVE: This study aimed to evaluate the extent to which the standardized physician competencies of the CanMEDS Framework are present in web-based patient reviews (WPRs) and to identify patients' perception of important physician qualities in the context of quality cancer care. METHODS: The WPRs of all university-affiliated medical oncologists in midsized cities with medical schools in the province of Ontario (Canada) were collected. Two reviewers (1 communication studies researcher and 1 health care professional) independently assessed the WPRs according to the CanMEDS Framework and identified common themes. Comment scores were then evaluated to identify κ agreement rates between the reviewers, and a descriptive quantitative analysis of the cohort was completed. Following the quantitative analysis, an inductive thematic analysis was performed. RESULTS: This study identified 49 actively practicing university-affiliated medical oncologists in midsized urban areas in Ontario. A total of 473 WPRs reviewing these 49 physicians were identified. Among the CanMEDS competencies, those defining the roles of medical experts, communicators, and professionals were the most prevalent (303/473, 64%; 182/473, 38%; and 129/473, 27%, respectively). Common themes in WPRs include medical skill and knowledge, interpersonal skills, and answering questions (from the patient to the physician). Detailed WPRs tend to include the following elements: experience and connection; discussion and evaluation of the physician's knowledge, professionalism, interpersonal skills, and punctuality; in positive reviews, the expression of feelings of gratitude and a recommendation; and in negative reviews, discouragement from seeking the physician's care. Patients' perception of medical skills is less specific than their perception of interpersonal qualities, although medical skills are the most commented-on element of care in WPRs. Patients' perception of interpersonal skills (listening, compassion, and overall caring demeanor) and other experiential phenomena, such as feeling rushed during appointments, is often specific and detailed. Details about a physician's interpersonal skills or "bedside manner" are highly perceived, valued, and shareable in an WPR context. A small number of WPRs reflected a distinction between the value of medical skills and that of interpersonal skills. The authors of these WPRs claimed that for them, a physician's medical skills and competence are more important than their interpersonal skills. CONCLUSIONS: CanMEDS roles and competencies that are explicitly patient facing (ie, those directly experienced by patients in their interactions with physicians and through the care that physicians provide) are the most likely to be present and reported on in WPRs. The findings demonstrate the opportunity to learn from WPRs, not simply to discern physicians' popularity but to grasp what patients may expect from their physicians. In this context, WPRs can represent a method for the measurement and assessment of patient-facing physician competency.

Barriers and Facilitators to Ophthalmology Visit Adherence in an Urban Hospital Setting.

Purpose: To explore barriers and facilitators to completing scheduled outpatient appointments at an urban academic hospital-based ophthalmology department. Methods: Potential participants were stratified by neighborhood Social Vulnerability Index (SVI) (range, 0-1.0, higher scores indicate greater vulnerability), and semistructured interviews were conducted with individuals 18 years and older with an SVI of greater than 0.61 (n = 17) and providers delivering care in the General Eye Clinic of the University of Illinois Chicago (n = 8). Qualitative analysis informed by human-centered design methods was conducted to classify barriers and facilitators into three domains of the Consolidated Framework for Implementation Research: outer setting, inner setting, and characteristics of individuals. Results: There were four main themes-transportation, time burden, social support, and economic situation-all of which were within the outer setting of the Consolidated Framework for Implementation Research; transportation was most salient. Although providers perceived health literacy as a barrier affecting motivation, patients expressed a high motivation to attend visits and felt well-educated about their condition. Conclusions: A lack of resources outside of the health system presents significant barriers for patients from neighborhoods with high SVI. Future efforts to improve adherence should focus on resource-related interventions in the outer setting. Improving access to eye care will require community-level interventions, particularly transportation. Translational Relevance: Understanding the barriers and facilitators within the Consolidated Framework for Implementation Research provides useful guidance for future interventions, specifically to focus future efforts to improve adherence on resource-related interventions.

Collagen Matrices Mediate Glioma Cell Migration Induced by an Electrical Signal.

Glioma cells produce an increased amount of collagen compared with normal astrocytes. The increasing amount of collagen in the extracellular matrix (ECM) modulates the matrix structure and the mechanical properties of the microenvironment, thereby regulating tumor cell invasion. Although the regulation of tumor cell invasion mainly relies on cell-ECM interaction, the electrotaxis of tumor cells has attracted great research interest. The growth of glioma cells in a three-dimensional (3D) collagen hydrogel creates a relevant tumor physiological condition for the study of tumor cell invasion. In this study, we tested the migration of human glioma cells, fetal astrocytes, and adult astrocytes in a 3D collagen matrix with different collagen concentrations. We report that all three types of cells demonstrated higher motility in a low concentration of collagen hydrogel (3 mg/mL and 5 mg/mL) than in a high concentration of collagen hydrogel (10 mg/mL). We further show that human glioma cells grown in collagen hydrogels responded to direct current electric field (dcEF) stimulation and migrated to the anodal pole. The tumor cells altered their morphology in the gels to adapt to the anodal migration. The directedness of anodal migration shows a field strength-dependent response. EF stimulation increased the migration speed of tumor cells. This study implicates the potential role of an dcEF in glioma invasion and as a target of treatment.

The Role of Acetate Kinase in the Human Parasite Entamoeba histolytica.

The human parasite Entamoeba histolytica, which causes approximately 100 million cases of amoebic dysentery each year, relies on glycolysis as the major source of ATP production from glucose as it lacks a citric acid cycle and oxidative phosphorylation. Ethanol and acetate, the two major glycolytic end products for E. histolytica, are produced at a ratio of 2:1 under anaerobic conditions, creating an imbalance between NADH production and utilization. In this study we investigated the role of acetate kinase (ACK) in acetate production during glycolysis in E. histolytica metabolism. Analysis of intracellular and extracellular metabolites demonstrated that acetate levels were unaffected in an ACK RNAi cell line, but acetyl-CoA levels and the NAD+/NADH ratio were significantly elevated. Moreover, we demonstrated that glyceraldehyde 3-phosphate dehydrogenase catalyzes the ACK-dependent conversion of acetaldehyde to acetyl phosphate in E. histolytica. We propose that ACK is not a major contributor to acetate production, but instead provides a mechanism for maintaining the NAD+/NADH balance during ethanol production in the extended glycolytic pathway.