Hookworm-Derived Metabolites Suppress Pathology in a Mouse Model of Colitis and Inhibit Secretion of Key Inflammatory Cytokines in Primary Human Leukocytes.

Iatrogenic hookworm therapy shows promise for treating disorders that result from a dysregulated immune system, including inflammatory bowel disease (IBD). Using a murine model of trinitrobenzenesulfonic acid-induced colitis and human peripheral blood mononuclear cells, we demonstrated that low-molecular-weight metabolites derived from both somatic extracts (LMWM-SE) and excretory-secretory products (LMWM-ESP) of the hookworm, Ancylostoma caninum, display anti-inflammatory properties. Administration to mice of LMWM-ESP as well as sequentially extracted fractions of LMWM-SE using both methanol (SE-MeOH) and hexane-dichloromethane-acetonitrile (SE-HDA) resulted in significant protection against T cell-mediated immunopathology, clinical signs of colitis, and impaired histological colon architecture. To assess bioactivity in human cells, we stimulated primary human leukocytes with lipopolysaccharide in the presence of hookworm extracts and showed that SE-HDA suppressed ex vivo production of inflammatory cytokines. Gas chromatography-mass spectrometry (MS) and liquid chromatography-MS analyses revealed the presence of 46 polar metabolites, 22 fatty acids, and five short-chain fatty acids (SCFAs) in the LMWM-SE fraction and 29 polar metabolites, 13 fatty acids, and six SCFAs in the LMWM-ESP fraction. Several of these small metabolites, notably the SCFAs, have been previously reported to have anti-inflammatory properties in various disease settings, including IBD. This is the first report showing that hookworms secrete small molecules with both ex vivo and in vivo anti-inflammatory bioactivity, and this warrants further exploration as a novel approach to the development of anti-inflammatory drugs inspired by coevolution of gut-dwelling hookworms with their vertebrate hosts.

Revisiting the Ancylostoma Caninum Secretome Provides New Information on Hookworm-Host Interactions.

Hookworm infection is a major tropical parasitic disease affecting almost 500 million people worldwide. These soil-transmitted helminths can survive for many years in the intestine of the host, where they feed on blood, causing iron deficiency anemia and other complications. These parasites release a variety of molecules known as excretory/secretory products (ESPs) that are involved in many different biological processes that govern parasite survival. Using a combination of separation techniques such as SDS-PAGE and OFFGEL electrophoresis, in combination with state-of-the-art mass spectrometry we have reanalyzed the dog hookworm, Ancylostoma caninum, ESPs (AcESP). We identified 315 proteins present in the AcESP, compared with just 105 identified in previous studies. The most highly represented family of proteins is the SCP/TAPs (110 of the 315 proteins), and the most abundant constituents of AcESP are homologues of the tissue inhibitors of metalloproteases (TIMP) family. Interestingly, we identified new homologs of well-known vaccine candidates and immunomodulatory proteins. This study provides novel information about the proteins secreted by A. caninum, and constitutes a comprehensive dataset to study the proteins involved in host-hookworm interactions.

Incidentally Detected Lung Cancer in Persons Too Young or Too Old for Lung Cancer Screening in a Mississippi Delta Cohort.

INTRODUCTION: Lung cancer risk in screening age-ineligible persons with incidentally detected lung nodules is poorly characterized. We evaluated lung cancer risk in two age-ineligible Lung Nodule Program (LNP) cohorts. METHODS: Prospective observational study comparing 2-year cumulative lung cancer diagnosis risk, lung cancer characteristics, and overall survival between low-dose computed tomography (LDCT) screening participants aged 50 to 80 years and LNP participants aged 35 to younger than 50 years (young) and older than 80 years (elderly). RESULTS: From 2015 to 2022, lung cancer was diagnosed in 329 (3.43%), 39 (1.07%), and 172 (6.87%) LDCT, young, and elderly LNP patients, respectively. The 2-year cumulative incidence was 3.0% (95% confidence intervals [CI]: 2.6%-3.4%) versus 0.79% (CI: 0.54%-1.1%) versus 6.5% (CI: 5.5%-7.6%), respectively, but lung cancer diagnosis risk was similar between young LNP and Lung CT Screening Reporting and Data System (Lung-RADS) 1 (adjusted hazard ratio [aHR] = 0.88 [CI: 0.50-1.56]) and Lung-RADS 2 (aHR = 1.0 [0.58-1.72]). Elderly LNP risk was greater than Lung-RADS 3 (aHR = 2.34 [CI: 1.50-3.65]), but less than 4 (aHR = 0.28 [CI: 0.22-0.35]). Lung cancer was stage I or II in 62.92% of LDCT versus 33.33% of young (p = 0.0003) and 48.26% of elderly (p = 0.0004) LNP cohorts; 16.72%, 41.03%, and 29.65%, respectively, were diagnosed at stage IV. The aggregate 5-year overall survival rates were 57% (CI: 48-67), 55% (CI: 39-79), and 24% (CI: 15-40) (log-rank p < 0.0001). Results were similar after excluding persons with any history of cancer. CONCLUSIONS: LNP modestly benefited persons too young or old for screening. Differences in clinical characteristics and outcomes suggest differences in biological characteristics of lung cancer in these three patient cohorts.

Program-Based Lung Cancer Care: A Prospective Observational Tumor Registry Linkage Study.

Introduction: Low-dose computed tomography screening (LDCT) and lung nodule programs (LNP) promote early lung cancer detection, improve survival; Multidisciplinary Care Programs (MDC) promote guideline-concordant care. The impact of such program-based care on "real-world" lung cancer survival is unquantified. We evaluated outcomes of lung cancer care delivered through structured programs in a community health care system. Methods: We conducted a cohort study linking institutional prospective observational LDCT, LNP and MDC databases with Tumor Registry of Baptist Cancer Center facilities. We categorized all patients diagnosed with lung cancer between 2011 and 2021 into program-based care versus non-program-based care cohorts. We compared patient characteristics, stage distribution, treatment modalities, survival and mortality in each pathway of care. Results: Of 12,148 patients, 237, 1,165, 1,140 and 9,606 were diagnosed through the LDCT, LNP, MDC or no program, respectively; non-program-based care sequentially diminished from 96.3% to 66.5%, diagnosis through LDCT increased from 0.5% to 7.1%, LNP from 3.5% to 20.8%; and MDC alone decreased from a high of 12.8% in 2014 to 5.6% in 2021. Program-based care was associated with earlier stage (p < 0.001), higher surgical resection rates (p < 0.001), greater use of adjuvant therapy (p < 0.001), better aggregate and stage-stratified survival (p < 0.001), and lower all-cause and lung cancer-specific mortality (p < 0.001). Recipients of non-program-based care were considerably less likely to receive lung cancer treatment; results remained consistent when patients receiving no treatment were excluded. Conclusions: Program-based care was associated with substantially better survival. Increasing access to program-based care should be explored as a matter of urgent public policy.

Evaluation of Lung Cancer Risk Among Persons Undergoing Screening or Guideline-Concordant Monitoring of Lung Nodules in the Mississippi Delta.

Importance: Guideline-concordant management of lung nodules promotes early lung cancer diagnosis, but the lung cancer risk profile of persons with incidentally detected lung nodules differs from that of screening-eligible persons. Objective: To compare lung cancer diagnosis hazard between participants receiving low-dose computed tomography screening (LDCT cohort) and those in a lung nodule program (LNP cohort). Design, Setting, and Participants: This prospective cohort study included LDCT vs LNP enrollees from January 1, 2015, to December 31, 2021, who were seen in a community health care system. Participants were prospectively identified, data were abstracted from clinical records, and survival was updated at 6-month intervals. The LDCT cohort was stratified by Lung CT Screening Reporting and Data System as having no potentially malignant lesions (Lung-RADS 1-2 cohort) vs those with potentially malignant lesions (Lung-RADS 3-4 cohort), and the LNP cohort was stratified by smoking history into screening-eligible vs screening-ineligible groups. Participants with prior lung cancer, younger than 50 years or older than 80 years, and lacking a baseline Lung-RADS score (LDCT cohort only) were excluded. Participants were followed up to January 1, 2022. Main Outcomes and Measures: Comparative cumulative rates of lung cancer diagnosis and patient, nodule, and lung cancer characteristics between programs, using LDCT as a reference. Results: There were 6684 participants in the LDCT cohort (mean [SD] age, 65.05 [6.11] years; 3375 men [50.49%]; 5774 [86.39%] in the Lung-RADS 1-2 and 910 [13.61%] in the Lung-RADS 3-4 cohorts) and 12 645 in the LNP cohort (mean [SD] age, 65.42 [8.33] years; 6856 women [54.22%]; 2497 [19.75%] screening eligible and 10 148 [80.25%] screening ineligible). Black participants constituted 1244 (18.61%) of the LDCT cohort, 492 (19.70%) of the screening-eligible LNP cohort, and 2914 (28.72%) of the screening-ineligible LNP cohort (P < .001). The median lesion size was 4 (IQR, 2-6) mm for the LDCT cohort (3 [IQR, 2-4] mm for Lung-RADS 1-2 and 9 [IQR, 6-15] mm for Lung-RADS 3-4 cohorts), 9 (IQR, 6-16) mm for the screening-eligible LNP cohort, and 7 (IQR, 5-11) mm for the screening-ineligible LNP cohort. In the LDCT cohort, lung cancer was diagnosed in 80 participants (1.44%) in the Lung-RADS 1-2 cohort and 162 (17.80%) in the Lung-RADS 3-4 cohort; in the LNP cohort, it was diagnosed in 531 (21.27%) in the screening-eligible cohort and 447 (4.40%) in the screening-ineligible cohort. Compared with Lung-RADS 1-2, the fully adjusted hazard ratios (aHRs) were 16.2 (95% CI, 12.7-20.6) for the screening-eligible cohort and 3.8 (95% CI, 3.0-5.0) for the screening-ineligible cohort; compared with Lung-RADS 3-4, the aHRs were 1.2 (95% CI, 1.0-1.5) and 0.3 (95% CI, 0.2-0.4), respectively. The stage of lung cancer was I to II in 156 of 242 patients (64.46%) in the LDCT cohort, 276 of 531 (52.00%) in the screening-eligible LNP cohort, and 253 of 447 (56.60%) in the screening-ineligible LNP cohort. Conclusions and Relevance: In this cohort study, the cumulative lung cancer diagnosis hazard of screening-age persons enrolled in the LNP was higher than that in a screening cohort, irrespective of smoking history. The LNP provided access to early detection for a higher proportion of Black persons.

Potential Impact of Criteria Modifications on Race and Sex Disparities in Eligibility for Lung Cancer Screening.

INTRODUCTION: Low-dose computed tomography (LDCT) screening reduces lung cancer mortality, but current eligibility criteria underestimate risk in women and racial minorities. We evaluated the impact of screening criteria modifications on LDCT eligibility and lung cancer detection. METHODS: Using data from a Lung Nodule Program, we compared persons eligible for LDCT by the following: U.S. Preventive Services Task Force (USPSTF) 2013 criteria (55-80 y, ≥30 pack-years of smoking, and ≤15 y since cessation); USPSTF2021 criteria (50-80 y, ≥20 pack-years of smoking, and ≤15 y since cessation); quit duration expanded to less than or equal to 25 years (USPSTF2021-QD25); reducing the pack-years of smoking to more than or equal to 10 years (USPSTF2021-PY10); and both (USPSTF2021-QD25-PY10). We compare across groups using the chi-square test or analysis of variance. RESULTS: The 17,421 individuals analyzed were of 56% female sex, 69% white, 28% black; 13% met USPSTF2013 criteria; 17% USPSTF2021; 18% USPSTF2021-QD25; 19% USPSTF2021-PY10; and 21% USPSTF2021-QD25-PY10. Additional eligible individuals by USPSTF2021 (n = 682) and USPSTF2021-QD25-PY10 (n = 1402) were 27% and 29% black, both significantly higher than USPSTF2013 (17%, p < 0.0001). These additional eligible individuals were 55% (USPSTF2021) and 55% (USPSTF2021-QD25-PY10) of female sex, compared with 48% by USPSTF2013 (p < 0.05). Of 1243 persons (7.1%) with lung cancer, 22% were screening eligible by USPSTF13. USPSTF2021-QD25-PY10 increased the total number of persons with lung cancer by 37%. These additional individuals with lung cancer were of 57% female sex (versus 48% with USPSTF2013, p = 0.0476) and 24% black (versus 20% with USPSTF2013, p = 0.3367). CONCLUSIONS: Expansion of LDCT screening eligibility criteria to allow longer quit duration and fewer pack-years of exposure enriches the screening-eligible population for women and black persons.

Front squat data reproducibility collected with a triple-axis accelerometer.

The purpose of our study was to assess data reproducibility from 2 consecutive front squat workouts, spaced 1 week apart, performed by American college football players (n = 18) as they prepared for their competitive season. For each workout, our methods entailed the performance of 3-6 front squat repetitions per set at 55, 65, and 75% of subject's 1 repetition maximum (1RM) load. In addition, a fourth set was done at a heavier load, with a resistance equal to 80 and 83% of their 1RM values, for the first and second workouts, respectively. A triple-axis accelerometer was affixed to a barbell to quantify exercise performance. Per load, the accelerometer measures peak values for the following indices: force, velocity, and power. To assess data reproducibility, inter-workout comparisons were made for 12 performance indices with 4 statistical test-retest measures: intraclass correlation coefficients, coefficients of variation (CVs), and the SEM expressed in both absolute and relative terms. Current results show that the majority of performance indices exceeded intraclass correlation (0.75-0.80) and CV (10-15%) values previously deemed as acceptable levels of data reproducibility. The 2 indices with the greatest variability were power and velocity values obtained at 55% of the 1RM load; thus, it was concluded that higher movement rates at the lightest load were the most difficult aspect of front squat performance to repeat successfully over time. Our practical applications imply lighter loads, with inherently higher rates of barbell movement, yield lower data reproducibility values.

Anthropometry as a predictor of vertical jump heights derived from an instrumented platform.

The current study purpose examined the vertical height-anthropometry relationship with jump data obtained from an instrumented platform. Our methods required college-aged (n = 177) subjects to make 3 visits to our laboratory to measure the following anthropometric variables: height, body mass, upper arm length (UAL), lower arm length, upper leg length, and lower leg length. Per jump, maximum height was measured in 3 ways: from the subjects' takeoff, hang times, and as they landed on the platform. Standard multivariate regression assessed how well anthropometry predicted the criterion variance per gender (men, women, pooled) and jump height method (takeoff, hang time, landing) combination. Z-scores indicated that small amounts of the total data were outliers. The results showed that the majority of outliers were from jump heights calculated as women landed on the platform. With the genders pooled, anthropometry predicted a significant (p < 0.05) amount of variance from jump heights calculated from both takeoff and hang time. The anthropometry-vertical jump relationship was not significant from heights calculated as subjects landed on the platform, likely due to the female outliers. Yet anthropometric data of men did predict a significant amount of variance from heights calculated when they landed on the platform; univariate correlations of men's data revealed that UAL was the best predictor. It was concluded that the large sample of men's data led to greater data heterogeneity and a higher univariate correlation. Because of our sample size and data heterogeneity, practical applications suggest that coaches may find our results best predict performance for a variety of college-aged athletes and vertical jump enthusiasts.

Data reliability from an instrumented vertical jump platform.

A Vertec jump measurement and training system measures vertical jump heights but not additional variables that would reveal how the performance was achieved. Technology advances to equipment now include additional variables that elucidate how jump performance is achieved. However, acceptance of new jump-related equipment is predicated on the reliability of the vertical heights it measures in relation to those assessed by the Vertec. Thus, our study compared vertical jump height reliability data from a newly created instrumented platform to those concurrently derived from the Vertec. Methods required subjects (n = 105) to perform 2 jump trials separated by at least 2 days of rest. Trials began with a warm-up, followed by 3 to 5 maximal-effort jumps. The Vertec was placed directly over the platform so, as jumps occurred, subjects took off and landed on the instrumented device. At the jump apex subjects contacted the highest Vertec slapstick possible to assess maximum height attained. Four height measurements were derived from each jump: 3 platform-based calculations (from subject's take-off, hang time, and landing) and 1 Vertec. The platform-based calculations were compared to Vertec data to assess the reliability of the instrumented device. Intraclass correlation coefficient (0.90), coefficient of variation (17.3%), standard error of measurement (0.9 cm), and smallest real difference (3.7 cm) results showed heights calculated from platform take-offs were most reliable to Vertec values. It was concluded take-off from the platform yielded jump heights that are a viable alternative to those derived from the Vertec. Practical applications suggest coaches may use the platform to derive reliable vertical jump data in addition to other variables to better understand the performance of their athletes.

Data reproducibility from an inertial kinetic exercise machine.

An inertial kinetic exercise (IKE, Oconomowoc, WI, USA) machine was created that permits high-velocity workouts suited for injury rehabilitation and speed development in athletes. In addition, its design does not entail the movement of resistance against Earth's gravitational pull. Thus, the design and operation associated with the IKE may be incorporated into novel environments such as space flight. Yet before it can be used for such reasons, IKE data reproducibility must first be ascertained, which serves as the present study's purpose. Methods to assess data reproducibility required subjects (n = 42) to perform 2 IKE elbow flexor trials, each comprising two 60-second sets separated by a 90-second rest. Preceded by a familiarization session on the IKE, subjects were instructed to perform high-speed elbow flexor trial repetitions in good form. Instrumentation entailed an accelerometer and a computer interfaced with software. Maximum and average forces (MF, AF), as well as work (W), were examined for reproducibility with several test-retest measures. Results showed high reproducibility, yet intertrial variability was greater due in part to the length of time between exercise sessions. It was concluded that MF reproducibility was inherently less partly due to the nature of the variable, which entailed a single instantaneous expression over each 60-second set. In contrast, AF and W denoted a cumulative, and thus better overall, representation of total exercise performance. Based on our high reproducibility values, present IKE instrumentation methods appear to be appropriate. Practical applications suggest that coaches and therapists may want to employ an IKE as instrumented in the present study to derive reproducible performance data on people who may use the device under their supervision.

beta(2)-Adrenergic agonist administration and strength training.

beta(2)-Adrenergic agonists (beta(2)AA) produce myriad effects throughout the human body. Prescribed concurrently with theophylline for the bronchodilatory effects they offer in the treatment of asthma and chronic obstructive pulmonary disease, beta(2)AA actions include many beneficial and adverse changes when administered to animals at supraphysiological doses. Beneficial changes include improved musculoskeletal health and function, which can be maintained because adverse changes are reduced if oral beta(2)AA are given at therapeutic dosages in humans with concurrent resistive exercise administration. Combined oral beta(2)AA-resistive exercise treatments have been shown to produce gains in musculoskeletal health and function in numerous healthy and disuse atrophy human models. The mechanism(s) by which beta(2)AA exert their effects are a function of multiple factors, not the least of which includes the type of model receiving the drug treatment. Combined oral beta(2)AA-resistive exercise treatments in humans showed that adverse effects were greatly reduced when prudent and safer drug administration practices were employed (eg, screening subjects for tolerance before drug treatment). Results from human research trials suggest that administration of the combined treatment improves musculoskeletal function and performance with minimal health risk if proper precautions are followed. A related issue is administration of the combined treatment as an ergogenic aid to athletic performance. Given the results presented in this article, physicians should be wary of potential drug abuse and administer beta(2)AA only under appropriate circumstances when such a treatment is warranted.

Of dogs and hookworms: man's best friend and his parasites as a model for translational biomedical research.

We present evidence that the dog hookworm (Ancylostoma caninum) is underutilised in the study of host-parasite interactions, particularly as a proxy for the human-hookworm relationship. The inability to passage hookworms through all life stages in vitro means that adult stage hookworms have to be harvested from the gut of their definitive hosts for ex vivo research. This makes study of the human-hookworm interface difficult for technical and ethical reasons. The historical association of humans, dogs and hookworms presents a unique triad of positive evolutionary pressure to drive the A. caninum-canine interaction to reflect that of the human-hookworm relationship. Here we discuss A. caninum as a proxy for human hookworm infection and situate this hookworm model within the current research agenda, including the various 'omics' applications and the search for next generation biologics to treat a plethora of human diseases. Historically, the dog hookworm has been well described on a physiological and biochemical level, with an increasing understanding of its role as a human zoonosis. With its similarity to human hookworm, the recent publications of hookworm genomes and other omics databases, as well as the ready availability of these parasites for ex vivo culture, the dog hookworm presents itself as a valuable tool for discovery and translational research.

A medicinal plant compound, capnoidine, prevents the onset of inflammation in a mouse model of colitis.

ETHNOPHARMACOLOGICAL RELEVANCE: The traditional uses of Corydalis dubia, Ajania nubigena and Pleurospermum amabile in the Bhutanese traditional medicine for treating disorders related to inflammatory conditions and the in vitro anti-inflammatory activity of their crude extracts inspired the isolation and the investigation of anticolitic properties of four pure compounds. MATERIALS AND METHODS: Three medicinal plants were collected from Himalayan Mountains of Bhutan. Capnoidine and scoulerine were isolated from C. dubia, linalool oxide acetate from A. nubigena and isomyristicin from P. amabile using natural product isolation protocols. Four compounds were investigated for their anti-inflammatory activities against IBD-colitis using chemically induced (TNBS) mice model of colitis. Capnoidine conferred the best preliminary protection against TNBS-induced colitis in mice and we have conducted in-depth pharmacological investigation of this compound including clinical symptoms, pathological signs, cytokine profiles, histological structure and inflammasomes using relevant bioassay protocols. RESULTS: Capnoidine-treated mice had significantly: a) improved clinical symptoms (body weight loss, mobility, piloerection and faecal consistency); b) reduced colon pathology (adhesion, oedema, ulceration, and colon length); c) altered inflammatory cytokines profiles within the colons; d) reduced levels of p-IκB-α (Ser32) and p-NF-κB p65 (Ser536) and e) reduced histological inflammation in the colon when compared with mice administered TNBS only. CONCLUSION: Capnoidine presents as a potential new anti-inflammatory drug lead candidate for diseases where current standard-of-care often fails and is associated with major side effects. It also validates the traditional uses of C. dubia against inflammatory conditions and underlines the value of pursuing bioactive compounds derived from traditionally used ethnobotanical medicines.

Diterpenoid alkaloids of Aconitum laciniatum and mitigation of inflammation by 14-O-acetylneoline in a murine model of ulcerative colitis.

Aconitum laciniatum is used in Bhutanese traditional medicine for treating various chronic infections and inflammatory conditions. We carried out in-depth isolation and characterization of the phytochemicals from the root component and determined the anti-inflammatory effects of the isolated compounds against chemically-induced colitis in mice. Five diterpenoid alkaloids - pseudaconitine, 14-veratroylpseudaconine, 14-O-acetylneoline, neoline, and senbusine A - were isolated from A. laciniatum for the first time. Two of the alkaloids were tested for anti-inflammatory properties in the TNBS-induced colitis model in mice. Various parameters were measured to assess pathology including weight loss, clinical and macroscopic scores, histological structure and IFN-γ production in the gut. Of the two alkaloids tested, 14-O-acetylneoline showed significant protection against different parameters of colitic inflammation. Compared to control mice that received TNBS alone, mice treated with 14-O-acetylneoline experienced significantly less weight loss and had significantly lower clinical scores, macroscopic pathology and grades of histological inflammation. Moreover, colonic IFN-γ mRNA levels were significantly reduced in mice that received 14-O-acetylneoline compared to control mice that received TNBS alone. This alkaloid is now considered a novel anti-colitis drug lead compound.