Biomimetic Total Synthesis of (+)-Nocardioazine B and Analogs.

Nocardioazines A and B are prenylated, bioactive pyrroloindoline natural products, isolated from Nocardiopsis, with a desymmetrized cyclo-d-Trp-d-Trp DKP core. Based on our deeper biosynthetic understanding, a biomimetic total synthesis of (+)-nocardioazine B is accomplished in merely seven steps and 23.2% overall yield. This pathway accesses regio- and stereoselectively C3-isoprenylated analogs of (+)-nocardioazine B, using the same number of steps and in similar efficiency. The successful strategy mandated that the biomimetic C3-prenylation step be executed early. The use of an unprotected carboxylic acid of Trp led to high diastereoselectivity toward formation of key intermediates exo-12a, exo-12b, and exo-12c (>19:1). Evidence shows that N1-methylation causes the prenylation reaction to bifurcate away to result in a C2-normal-prenylated isomer. Nocardioazine A, possessing an isoprenoidal-epoxide bridge, inhibits P-glycoprotein (P-gp)-mediated membrane efflux, in multidrug-resistant mammalian colon cancer cells. As several P-gp inhibitors have failed due to their toxicity effects, endogenous amino-acid-derived noncytotoxic inhibitors (from the nocardioazine core) are worthy leads toward a rejuvenated strategy against resistant carcinomas. This total synthesis provides direct access to Trp-derived isoprenylated DKP natural products and their derivatives.

Prevalence of Triad-RED-S symptoms in high-level Kenyan male and female distance runners and corresponding control groups.

PURPOSE: This study examined and compared select Triad-RED-S components/risk factors in high-level Kenyan male and female distance runners to corresponding control groups; focusing on examining energy intake (EI), bone indices, and hormonal markers. METHODS: A cross-sectional, observational design was used in which Kenyan male and female (n = 30 and n = 26, respectively) middle- and long-distance runners and corresponding male and female control groups (n = 29 and n = 29, respectively) were examined. The participant's bone mineral density (BMD) at the lumbar spine, right femur, and total body were measured using a dual-energy X-ray absorptiometry analysis. Complete blood counts (CBC) were done on the whole blood specimens and hormonal measurements were performed on plasma specimens. In addition, athletes completed metabolic testing to determine maximal oxygen uptakes and 7-day dietary diaries. RESULTS: Overall daily EI across runners and controls within each sex were low, but not significantly different (p > 0.05). Prevalence of low BMD values (Z score < - 2.0) was comparable across groups in each sex (p > 0.05). CBC measures suggested that both runners and controls were healthy. Finally, slight hormonal differences between runners and their respective controls existed (p < 0.05), but were not clinically meaningful or observed in typical Triad-RED-S-related parameters. CONCLUSION: High-level Kenyan male and female runners had low daily EI, but no tendency toward a higher prevalence of low BMD, or Triad-RED-S-related hormonal abnormalities. The occurrence of low EI was not a major risk factor in our athletes; this calls into question whether the current criteria for Triad-RED-S are entirely applicable for athletes of African ethnicity.

Shorter Ground Contact Time and Better Running Economy: Evidence From Female Kenyan Runners.

ABSTRACT: Mooses, M, Haile, DW, Ojiambo, R, Sang, M, Mooses, K, Lane, AR, and Hackney, AC. Shorter ground contact time and better running economy: evidence from female Kenyan runners. J Strength Cond Res 35(2): 481-486, 2021-Previously, it has been concluded that the improvement in running economy (RE) might be considered as a key to the continued improvement in performance when no further increase in V̇o2max is observed. To date, RE has been extensively studied among male East African distance runners. By contrast, there is a paucity of data on the RE of female East African runners. A total of 10 female Kenyan runners performed 3 × 1,600-m steady-state run trials on a flat outdoor clay track (400-m lap) at the intensities that corresponded to their everyday training intensities for easy, moderate, and fast running. Running economy together with gait characteristics was determined. Subjects showed moderate to very good RE at the first (202 ± 26 ml·kg-1·km-1) and second (188 ± 12 ml·kg-1·km-1) run trials, respectively. Correlation analysis revealed significant relationship between ground contact time (GCT) and RE at the second run (r = 0.782; p = 0.022), which represented the intensity of anaerobic threshold. This study is the first to report the RE and gait characteristics of East African female athletes measured under everyday training settings. We provided the evidence that GCT is associated with the superior RE of the female Kenyan runners.

Correction: The expanding spectrum of diketopiperazine natural product biosynthetic pathways containing cyclodipeptide synthases.

Correction for 'The expanding spectrum of diketopiperazine natural product biosynthetic pathways containing cyclodipeptide synthases' by Paul Borgman et al., Org. Biomol. Chem., 2019, DOI: 10.1039/c8ob03063d.

The expanding spectrum of diketopiperazine natural product biosynthetic pathways containing cyclodipeptide synthases.

Microorganisms are remarkable chemists, with enzymes as their tools for executing multi-step syntheses to yield myriad natural products. Microbial synthetic aptitudes are illustrated by the structurally diverse 2,5-diketopiperazine (DKP) family of bioactive nonribosomal peptide natural products. Nonribosomal peptide synthetases (NRPSs) have long been recognized as catalysts for formation of DKP scaffolds from two amino acid substrates. Cyclodipeptide synthases (CDPSs) are more recently recognized catalysts of DKP assembly, employing two aminoacyl-tRNAs (aa-tRNAs) as substrates. CDPS-encoding genes are typically found in genomic neighbourhoods with genes encoding additional biosynthetic enzymes. These include oxidoreductases, cytochrome P450s, prenyltransferases, methyltransferases, and cyclases, which equip the DKP scaffold with groups that diversify chemical structures and confer biological activity. These tailoring enzymes have been characterized from nine CDPS-containing biosynthetic pathways to date, including four during the last year. In this review, we highlight these nine DKP pathways, emphasizing recently characterized tailoring reactions and connecting new developments to earlier findings. Featured pathways encompass a broad spectrum of chemistry, including the formation of challenging C-C and C-O bonds, regioselective methylation, a unique indole alkaloid DKP prenylation strategy, and unprecedented peptide-nucleobase bond formation. These CDPS-containing pathways also provide intriguing models of metabolic pathway evolution across related and divergent microorganisms, and open doors to synthetic biology approaches for generation of DKP combinatorial libraries. Further, bioinformatics analyses support that much unique genetically encoded DKP tailoring potential remains unexplored, suggesting opportunities for further expansion of Nature's biosynthetic spectrum. Together, recent studies of DKP pathways demonstrate the chemical ingenuity of microorganisms, highlight the wealth of unique enzymology provided by bacterial biosynthetic pathways, and suggest an abundance of untapped biosynthetic potential for future exploration.

Prevalence of Low Energy Availability in Competitively Trained Male Endurance Athletes.

Background and Objectives: Relative energy deficiency in sport (RED-S) has been introduced as a broad-spectrum syndrome leading to possible dysfunction in numerous physiological systems, driven primarily by low energy availability (EA). Research in females has identified specific EA cut-points indicative of risk level for developing physiological and performance disturbances. Cut-points in males have yet to be evaluated. This study examined the prevalence of low EA in competitive (non-elite), recreationally trained (CRT) male endurance athletes. Materials and Methods: Subjects were 108 CRT (38.6 ± 13.8 y; 12.2 ± 5.4 h/wk training) male endurance athletes (runners, cyclists, triathletes) who completed a descriptive survey online via Qualtrics® and returned 3 day diet and exercise training records. EA was calculated from returned surveys and training records. Resting metabolic rate (RMR) and lean body mass (LBM) were estimated from self-reported survey data. Prevalence of risk group was categorized based on the female cut-points: at risk (AR) ≤30 kcal/kg LBM, moderate risk (MR) = 30-45 kcal/kg LBM, or no risk (NR) ≥45 kcal/kg LBM. Results: In this sample, 47.2% (n = 51) were classified as AR, 33.3% (n = 36) as MR, and 19.4% (n = 21) as NR for low EA. Cyclists had lower EA (26.9 ± 17.4 kcal/kg LBM, n = 45) than runners (34.6 ± 13.3 kcal/kg LBM, n = 55, p = 0.016) and all other sport categories (39.5 ± 19.1 kcal/kg LBM, n = 8, p = 0.037). Conclusions: The findings indicate this sample had a high prevalence of risk for low EA, at 47.2%. Only 19.4% of participants were at no risk, meaning ~80% of participants were at some degree of risk of experiencing low EA. Cyclists were at greater risk in this cohort of low EA, although why this occurred was unclear and is in need of further investigation. Future research should address whether the current female cut-points for low EA are appropriate for use in male populations.

Intranasal mast cell tumor in the dog: A case series.

The medical records of 4 dogs with histologically confirmed intranasal mast cell tumors (MCTs) were retrospectively evaluated to determine their biological behavior. Information on signalment, presenting clinical signs, tumor grade, treatment administered, and survival times was obtained from the medical record. All 4 patients had high grade tumors and received chemotherapy. Survival times ranged from 27 to 134 days. All 4 dogs showed signs of local or distant disease progression, suggestive of an aggressive behavior of intranasal MCTs.

Tumeur mastocytaire intranasale chez le chien : une série de cas. Les dossiers médicaux de quatrechiens qui avaient eu des tumeurs mastocytaires intranasales confirmées par histologie ont été rétrospectivement évalués afin de déterminer leur comportement biologique. Des renseignements sur le signalement, les signes cliniques de présentation, le grade de la tumeur, le traitement administré et les temps de survie ont été obtenus dans le dossier médical. Les quatre patients avaient des tumeurs de grade élevé et ont reçu de la chimiothérapie. Les temps de survie ont varié de 27 à 134 jours. Les quatre chiens ont manifesté des signes de progression locale ou distante de la maladie, suggérant un comportement agressif des tumeurs mastocytaires intranasales.(Traduit par Isabelle Vallières).

Cortisol and testosterone dynamics following exhaustive endurance exercise.

PURPOSE: Cortisol (C) and testosterone (T) are impacted significantly by prolonged endurance exercise with inverse responses. Increases in C are witnessed concurrently with decrements in T, possibly impacting recovery. This study was conducted to assess the dynamics of C and free T (fT) concentration and recovery time following an exhaustive endurance exercise session (EES). METHODS: 12 endurance-trained males (X ± SD: VO2max 66.3±4.8 ml/kg/min, age 22.8 ± 3.1 years, body fat 11.0 ± 1.4 %, training 7.1 ± 3.2 years) completed a treadmill EES at ventilatory threshold (74.7 ± 4.6 % of VO2max; 96.9 ± 10.8 min). Basal blood C and fT were collected at baseline: -48, -24 h, and immediately before (0 h) the EES as well as immediately (+0 h), +24 h, +48 h, and +72 h after the EES. Blood glucose (G) was measured to confirm no undue influence on C. Statistically data were analyzed with repeated measures ANOVA (LSD post hoc). RESULTS: C (nmol/L) increased significantly from -48 h (321 ± 59) to +0 h (701 ± 178) (p < 0.001), and displayed a baseline overshoot with +24 h (209 ± 67) being significantly lower than -48 and +0 h (p < 0.03). fT (pmol/L) decreased significantly from -48 h (161 ± 40) to +0 h (106 ± 38) (p < 0.01) and remained lower at +24 h (110 ± 33) and +48 h (129 ± 30) (p < 0.001). G remained stable throughout. A moderately negative correlation (r = -0.636, p < 0.026) was found between C and fT at +0 h. CONCLUSIONS: EES recovery may require 48 h for C and 72 h for fT to return to baseline values. Furthermore, C and fT were only correlated immediately post-exercise. Future research should perform more frequent measurements throughout time course.

Characterization of the nocardiopsin biosynthetic gene cluster reveals similarities to and differences from the rapamycin and FK-506 pathways.

Macrolide-pipecolate natural products, such as rapamycin (1) and FK-506 (2), are renowned modulators of FK506-binding proteins (FKBPs). The nocardiopsins, from Nocardiopsis sp. CMB-M0232, are the newest members of this structural class. Here, the biosynthetic pathway for nocardiopsins A-D (4-7) is revealed by cloning, sequencing, and bioinformatic analyses of the nsn gene cluster. In vitro evaluation of recombinant NsnL revealed that this lysine cyclodeaminase catalyzes the conversion of L-lysine into the L-pipecolic acid incorporated into 4 and 5. Bioinformatic analyses supported the conjecture that a linear nocardiopsin precursor is equipped with the hydroxy group required for macrolide closure in a previously unobserved manner by employing a P450 epoxidase (NsnF) and limonene epoxide hydrolase homologue (NsnG). The nsn cluster also encodes candidates for tetrahydrofuran group biosynthesis. The nocardiopsin pathway provides opportunities for engineering of FKBP-binding metabolites and for probing new enzymology in nature's polyketide tailoring arsenal.

Correction: Synergism between genome sequencing, tandem mass spectrometry and bio-inspired synthesis reveals insights into nocardioazine B biogenesis.

Correction for 'Synergism between genome sequencing, tandem mass spectrometry and bio-inspired synthesis reveals insights into nocardioazine B biogenesis' by Norah Alqahtani et al., Org. Biomol. Chem., 2015, 13, 7177-7192.

Synergism between genome sequencing, tandem mass spectrometry and bio-inspired synthesis reveals insights into nocardioazine B biogenesis.

Marine actinomycete-derived natural products continue to inspire chemical and biological investigations. Nocardioazines A and B (3 and 4), from Nocardiopsis sp. CMB-M0232, are structurally unique alkaloids featuring a 2,5-diketopiperazine (DKP) core functionalized with indole C3-prenyl as well as indole C3- and N-methyl groups. The logic of their assembly remains cryptic. Bioinformatics analyses of the Nocardiopsis sp. CMB-M0232 draft genome afforded the noz cluster, split across two regions of the genome, and encoding putative open reading frames with roles in nocardioazine biosynthesis, including cyclodipeptide synthase (CDPS), prenyltransferase, methyltransferase, and cytochrome P450 homologs. Heterologous expression of a twelve gene contig from the noz cluster in Streptomyces coelicolor resulted in accumulation of cyclo-l-Trp-l-Trp DKP (5). This experimentally connected the noz cluster to indole alkaloid natural product biosynthesis. Results from bioinformatics analyses of the noz pathway along with challenges in actinomycete genetics prompted us to use asymmetric synthesis and mass spectrometry to determine biosynthetic intermediates in the noz pathway. The structures of hypothesized biosynthetic intermediates 5 and 12-17 were firmly established through chemical synthesis. LC-MS and MS-MS comparison of these synthetic compounds with metabolites present in chemical extracts from Nocardiopsis sp. CMB-M0232 revealed which of these hypothesized intermediates were relevant in the nocardioazine biosynthetic pathway. This established the early and mid-stages of the biosynthetic pathway, demonstrating that Nocardiopsis performs indole C3-methylation prior to indole C3-normal prenylation and indole N1'-methylation in nocardioazine B assembly. These results highlight the utility of merging bioinformatics analyses, asymmetric synthetic approaches, and mass spectrometric metabolite profiling in probing natural product biosynthesis.

Pre-exercise participation cardiovascular screening in a heterogeneous cohort of adult cancer patients.

BACKGROUND: The purpose of this study was to investigate the extent of pre-exercise participation ("preparticipation") health screening in a heterogeneous cohort of adult cancer patients. METHODS: Patients (n = 413) with histologically confirmed solid or hematologic malignancy were categorized into preparticipation health screening risk stratification based on American College Sports Medicine (ACSM) recommendations. Risk of an exercise-related event was evaluated during a symptom-limited cardiopulmonary exercise test (CPET) with 12-lead electrocardiography (ECG). RESULTS: Participant risk was categorized as low risk (n = 59, 14%), moderate risk (n = 217, 53%), and high risk (n = 137, 33%). Mean peak oxygen consumption was 21.7 ± 6.7 mL/kg(-1) per minute(-1) or 19.5 ± 21.7% below age- and sex-predicted sedentary values. No major serious adverse events or fatal events were observed during CPET procedures. A total of 31 positive ECG tests were observed, for an event rate of 8%. ACSM risk stratification did not predict the risk of a positive test. Age, statin use, antiplatelet therapy use, cardiovascular disease, prior treatment with anthracycline or radiation therapy, and being sedentary were predictors of a positive test (all p < .10). CONCLUSION: The patient risk-stratification profile strongly suggests that the use of formalized preparticipation health screening is required in all oncology scenarios; however, risk of an exercise-induced event is low, suggesting that the use of exercise testing is not required for pre-exercise clearance in the majority of patients.

Utility of 3-dimensional echocardiography, global longitudinal strain, and exercise stress echocardiography to detect cardiac dysfunction in breast cancer patients treated with doxorubicin-containing adjuvant therapy.

Conventional resting left ventricular ejection fraction (LVEF) assessments have limitations for detecting doxorubicin (DOX)-related cardiac dysfunction. Novel resting echocardiographic parameters, including 3-dimensional echocardiography (3DE) and global longitudinal strain (GLS), have potential for early identification of chemotherapy-related myocardial injury. Exercise "stress" is an established method to uncover impairments in cardiac function but has received limited attention in the adult oncology setting. We evaluated the utility of an integrated approach using 3DE, GLS, and exercise stress echocardiography for detecting subclinical cardiac dysfunction in early breast cancer patients treated with DOX-containing chemotherapy. Fifty-seven asymptomatic women with early breast cancer (mean 26 ± 22 months post-chemotherapy) and 20 sex-matched controls were studied. Resting left ventricular (LV) function was assessed by LVEF using 2-dimensional echocardiography (2DE) and 3DE and by GLS using 2-dimensional speckle-tracking echocardiography (2D-STE). After resting assessments, subjects completed cardiopulmonary exercise testing with stress 2DE. Resting LVEF was lower in patients than controls by 3DE (55 ± 4 vs. 59 ± 5 %; p = 0.005) but not 2DE (56 ± 4 vs. 58 ± 3 %; p = 0.169). 10 of 51 (20 %) patients had GLS greater than or equal to -17 %, which was below the calculated lower limit of normal (control mean 2SD); this patient subgroup had a mean 20 % impairment in GLS (-16.1 ± 0.9 vs. -20.1 ± 1.5 %; p < 0.001), despite similar LVEF by 2DE and 3DE compared to controls (p > 0.05). Cardiopulmonary function (VO2peak) was 20 % lower in patients than controls (p < 0.001). Exercise stress 2DE assessments of stroke volume (61 ± 11 vs. 69 ± 15 ml; p = 0.018) and cardiac index (2.3 ± 0.9 vs. 3.1 ± 0.8 l min(-1) m(-2) mean increase; p = 0.003) were lower in patients than controls. Post-exercise increase in cardiac index predicted VO2peak (r = 0.429, p = 0.001). Resting 3DE, GLS, and exercise stress 2DE detect subclinical cardiac dysfunction not apparent with resting 2DE in post-DOX breast cancer patients.

Safety and efficacy of aerobic training in operable breast cancer patients receiving neoadjuvant chemotherapy: a phase II randomized trial.

BACKGROUND: To evaluate the safety and efficacy of moderate-to-high intensity aerobic training in breast cancer patients receiving neoadjuvant chemotherapy. METHODS: Twenty patients with stage IIB-IIIC operable breast cancer were randomly assigned to receive doxorubicin plus cyclophosphamide (AC) or AC in combination with aerobic training (AC + AET) (n = 10/group) for 12 weeks. The AC+ AET group performed three supervised aerobic cycle ergometry sessions per week at 60%-100% of exercise capacity (VO2peak). Safety outcomes included exercise testing as well as treatment- and exercise training-related adverse events (AEs), whereas efficacy outcomes included cardiopulmonary function and patient-reported outcomes (PROs) as measured by a cardiopulmonary exercise test (CPET) and Functional Assessment of Cancer Therapy-Breast (FACT-B) scale. RESULTS: Twelve non-significant ECG abnormalities and three non-life threatening events occurred during CPET procedures. One AE was reported during aerobic training. There were no significant between group differences for clinician-documented events (e.g. pain, nausea) or hematological parameters (p's > 0.05). Attendance and adherence rates to aerobic training were 82% and 66%, respectively. Intention-to-treat analysis indicated that VO2peak increased by 2.6 ± 3.5 ml/kg/min (+ 13.3%) in the AC + AET group and decreased by 1.5 ± 2.2 ml/kg/min (-8.6%) in the AC group (between group difference, p = 0.001). FACT-B increased 11.1 points in the AC + AET group compared to a 1.5 point decrease in the AC group (between group difference, p = 0.685). CONCLUSION: Moderate-to-high intensity aerobic training when conducted with one-on-one supervision is a safe adjunct therapy associated with improvements in cardiopulmonary function and select PROs during neoadjuvant chemotherapy.

Characterization of an orphan diterpenoid biosynthetic operon from Salinispora arenicola.

While more commonly associated with plants than microbes, diterpenoid natural products have been reported to have profound effects in marine microbe-microbe interactions. Intriguingly, the genome of the marine bacterium Salinispora arenicola CNS-205 contains a putative diterpenoid biosynthetic operon, terp1. Here recombinant expression studies are reported, indicating that this three-gene operon leads to the production of isopimara-8,15-dien-19-ol (4). Although 4 is not observed in pure cultures of S. arenicola, it is plausible that the terp1 operon is only expressed under certain physiologically relevant conditions such as in the presence of other marine organisms.

Driver rehabilitation programs: defining program models, services, and expertise.

This paper highlights the critical need for a diverse span of services targeted at older drivers that is clear and understandable to health care professionals, service providers, and consumers. The paper describes how a panel of expert driver rehabilitation specialists and researchers on older drivers affirmed consensus statements addressing the need for clarification of terms and services. It also presents a new document that describes a spectrum of driver services from education to specific driver rehabilitation services. The document will provide consumers, referral sources, payers, and stakeholders invested in older drivers' services, with the information to refer the right people to the right service at the right time.

Influence of energy availability on metabolic hormonal profiles in east African female and male distance runners.

BACKGROUND: Energy availability (EA) and relative energy deficiency in sport (RED-S) are understudied in East African endurance athletes, both females (F) and males (M). This study assessed the metabolic hormonal profiles of such athletes relative to their EA status. METHODS: Forty athletes (F=16, M=24) had their EA status, training, maximal oxygen uptake, and resting blood samples assessed using standard research practices. Subjects were stratified into two groups, high EA (HiEA) and low EA (LoEA) based on combined median value. RESULTS: Cortisol (P=0.034) and insulin (P=0.044) were significantly elevated in the LoEA group, while growth hormone (P=0.045) was significantly suppressed; and, prolactin (P=0.078) trended towards suppression, respectively compared to the HiEA group. All other hormonal comparison were non-significant. CONCLUSIONS: Metabolic hormonal profiles of female and male African distance runners are affected by their EA status. Aspects of these alterations agree in part with published findings based upon White populations, although some differences exist and need further investigation.

Structures and comparative characterization of biosynthetic gene clusters for cyanosporasides, enediyne-derived natural products from marine actinomycetes.

Cyanosporasides are marine bacterial natural products containing a chlorinated cyclopenta[a]indene core of suspected enediyne polyketide biosynthetic origin. Herein, we report the isolation and characterization of novel cyanosporasides C-F (3-6) from the marine actinomycetes Salinispora pacifica CNS-143 and Streptomyces sp. CNT-179, highlighted by the unprecedented C-2' N-acetylcysteamine functionalized hexose group of 6. Cloning, sequencing, and mutagenesis of homologous ~50 kb cyanosporaside biosynthetic gene clusters from both bacteria afforded the first genetic evidence supporting cyanosporaside's enediyne, and thereby p-benzyne biradical, biosynthetic origin and revealed the molecular basis for nitrile and glycosyl functionalization. This study provides new opportunities for bioengineering of enediyne derivatives and expands the structural diversity afforded by enediyne gene clusters.