Feeding Studies Research Articles

An Integrated Module Performs Selective 'Online' Epoxidation in the Biosynthesis of the Antibiotic Mupirocin.

The delineation of the complex biosynthesis of the potent antibiotic mupirocin, which consists of a mixture of pseudomonic acids (PAs) isolated from Pseudomonas fluorescens NCIMB 10586, presents significant challenges, and the timing and mechanisms of several key transformations remain elusive. Particularly intriguing are the steps that process the linear backbone from the initial polyketide assembly phase to generate the first cyclic intermediate PA-B. These include epoxidation as well as incorporation of the tetrahydropyran (THP) ring and fatty acid side chain required for biological activity. Herein, we show that the mini-module MmpE performs a rare online (ACP-substrate) epoxidation and is integrated ('in-cis') into the polyketide synthase via a docking domain. A linear polyketide fragment with six asymmetric centres was synthesised using a convergent approach and used to demonstrate substrate flux via an atypical KS0 and a previously unannotated ACP (MmpE_ACP). MmpE_ACP-bound synthetic substrates were critical in demonstrating successful epoxidation in vitro by the purified MmpE oxidoreductase domain. Alongside feeding studies, these results confirm the timing as well as chain length dependence of this selective epoxidation. These mechanistic studies pinpoint the location and nature of the polyketide substrate prior to the key formation of the THP ring and esterification that generate PA-B.

Polyketide Origin of the Indole Ring during the Biosynthesis of Indole Alkaloid Coprisidins.

Coprisidins, a new class of indole alkaloids, feature a 1,4-naphthoquinone moiety attached to C-3 of 2-oxindole. Heterologous expression of a type II polyketide synthase-containing gene cluster resulted in the generation of three coprisidins. Gene disruption and isotope feeding studies suggested that the 2-oxindole moieties of coprisidins originate from a tetracyclic aromatic polyketide through oxidative rearrangements. This represents a novel biosynthetic route for the synthesis of indole rings in nature.

8548 Breast Development in Infant Girls: Does Endogenous Estrogen Production During Minipuberty Play a Role?

Abstract Disclosure: M. Goldberg: None. S. Zhao: None. A. Kelly: None. V.A. Stallings: None. D.M. Umbach: None. W.J. Rogan: None. N. Shaw: None. D.P. Sandler: None. Background: Breast tissue, present at birth in most term infants, regresses during infancy but persists longer in girls than boys. In girls, this persistence is thought to be due in part to stimulation by estradiol (E2) produced during minipuberty, the postnatal activation of the hypothalamic-pituitary-gonadal axis. Empirical data are sparse, however. Because E2 concentrations in infant girls fluctuate, repeated E2 measures may be necessary to adequately characterize production across minipuberty and to detect effects on estrogen-sensitive breast tissue. Hypothesis: Higher average exposure to E2 during minipuberty is associated with larger breast bud diameter in late infancy in term females. Methods: We leveraged data from 136 girls from the Infant Feeding and Early Development (IFED) Study, a prospective cohort of reproductive development in term infants enrolled during 2010-2013 from the Philadelphia area. We measured E2 in serum samples collected at study visits every 2-4 weeks from age 2-32 weeks. We assessed breast bud diameter via ultrasound at the 32-week visit. For data analysis, we log2-transformed both E2 and breast bud diameter. For E2 concentrations below the limit of detection (2.99 pg/mL, 35% of samples), we applied multiple imputation. We first calculated the intraclass correlation coefficient (ICC) of repeated E2 to understand the relative magnitude of total variation from within subjects. We then calculated subject-specific mean of E2 across time and estimated associations with breast bud diameter at the 32-week visit using both age- and multivariable-adjusted linear regression models. We present results as percent change in breast bud diameter per doubling in average E2 concentration ([2β-1] x 100%). Results: Most infants were Black (71%) and formula-fed (76%). The median number of E2 measures per girl was 9 (range: 5-10). Most of the variation in E2 concentrations was due to within-individual variation (ICC=0.27). The geometric mean E2 concentration from 2-32 weeks of age was 5.1 pg/mL (95% confidence interval [CI]: 4.7-5.4). The median breast bud diameter at the 32-week visit was 13.7mm (range: 4.4-30.5). A doubling in average E2 concentration across minipuberty was associated with a 26% larger breast bud diameter (95% CI 10-43%). This association was similar after adjustment for feeding method, race, gestational age, neonatal weight, and breast bud diameter at the neonatal visit (percent change: 28%, 95% CI: 12-46%). Conclusions: Our results suggest that endogenous estrogen production in females during minipuberty stimulates the infant breast bud. More research is needed to investigate the potential biologic relevance of this hormonal stimulation of infant breast tissue to later breast development and risk of malignant and benign breast disorders. Presentation: 6/1/2024

7814 Non-fasting and Fasting Mixed Meal Tolerance Tests Equally Capture the Triglyceride Response in Participants With and Without MASLD

Abstract Disclosure: L.A. Gonzalez Ramirez: None. Z. He: None. J.N. Binongo: None. H.E. Huneault: None. A. Ramirez Tovar: None. J.A. Welsh: None. M.B. Vos: Consulting Fee; Self; Boehringer Ingelheim, Prosciento, Target Real World Solutions, Intercept Pharmaceuticals, INC, Novo Nordisk. Grant Recipient; Self; Target Real World Solutions. Mixed meal tolerance tests (MMTTs) are functional assessments that may identify metabolic dysregulation earlier than fasting tests during the progression to metabolic dysfunction-associated steatotic liver disease (MASLD). Although standards to conduct MMTTs in adults are readily available, the knowledge gap for children is quite large. Furthermore, conducting this type of study in children is challenging because they are less tolerant of long fasting intervals, long periods of testing, and early morning visits that interrupt school attendance. The objective of this study was to assess if conducting an MMTT in a non-fasting state will equally identify variance in the metabolic response of triglycerides (TG) between healthy children and those living with MASLD when compared to a 12-hour fasting MMTT. We collected blood samples every two hours in our 2-day randomized crossover feeding study. During each visit, 9 children with MASLD and 10 healthy controls aged 10 to 18 y underwent 1 standardized MMTT after a 12-hour overnight fasting (breakfast), followed by 2 standardized MMTTs in a non-fasting state (lunch and dinner). Participants were randomized to receive a glucose or fructose beverage at visit 1 and the other at visit 2. For uniformity, we only used data from the glucose beverage visit. We found that TG levels were statistically higher in the MASLD participants when compared to the non-MASLD group at each time point (p= 0.01). The maximum TG metabolic response peak for each MMTT was higher and more delayed in the MASLD group compared to the non-MASLD group. Within groups, postprandial TG level patterns were not significantly different when comparing breakfast, lunch, and dinner. Although TG levels varied throughout the day, the pattern after every meal was similar. These findings suggest that it is possible to identify differences in the TG metabolic response between children living with MASLD and healthy participants in the postprandial state. The results from our study also support the use of non-fasting MMTTs when examining TG metabolism in children, though further research to validate these findings is needed. Presentation: 6/1/2024

Acceptability and tolerance study of a high protein semi-elemental feed containing phgg fibre

Acceptability and tolerance study of a high protein semi-elemental feed containing phgg fibre

Statement complementing the EFSA Scientific Opinion on application (EFSA-GMO-NL-2015-126) for authorisation of food and feed containing, consisting of and produced from genetically modified soybean MON 87705 × MON 87708 × MON 89788.

Following a request from the European Commission, the GMO Panel assessed additional information related to the application for authorisation of food and feed containing, consisting of and produced from genetically modified soybean MON × MON 87708 × MON 89788 (EFSA-GMO-NL-2015-126). The applicant conducted a 90-day feeding study on GM soybean MON 87705 and provided a proposal for post-market monitoring considering the altered fatty acid profile of GM soybean MON 87705 × MON 87708 × MON 89788, to fulfil the deficiencies identified by EFSA GMO Panel, addressing elements that remained inconclusive from a previous EFSA scientific opinion issued in 2020. The GMO Panel concludes that the 90-day feeding study on GM soybean MON 87705 is in line with the requirements of Regulation (EU) No 503/2013 and that no treatment-related adverse effects were observed in rats after feeding diets containing soybean MON 87705 meals at 30% or 15% for 90 days. The GMO Panel reiterates the recommendation for a PMM for food in accordance with Regulation (EC) No 1829/2003 and Regulation (EU) No 503/2013 and concludes that the proposal provided by the applicant is in line with the recommendations described for the PMM plan of soybean MON 87705 × MON 87708 × MON 89788 in the adopted scientific opinion. Taking into account the previous assessment and the new information, the GMO Panel concludes that soybean MON 87705 × MON 87708 × MON 89788, as assessed in the scientific opinion on application EFSA-GMO-NL-2015-126 and in the supplementary toxicity study, is as safe as its non-GM comparator and the non-GM reference varieties tested and does not represent a nutritional concern in humans and animals, within the scope of this application.

Structure and Biosynthesis of Perochalasins A-C, Open-Chain Merocytochalasans Produced by the Marine-Derived Fungus Peroneutypa sp. M16.

Novel open-chain merocytochalasans, perochalasins A-C (1-3), containing an unusual N-O six-membered heterocyclic moiety, were isolated from cultures of the marine-derived Peroneutypa sp. M16 fungus, along with cytochalasin Z27 (4), cytochalasin Z28 (5), [12]-cytochalasin (6), and phenochalasin B (7). The structures of compounds 1-3 were established by analysis of the spectroscopic data. Full genome sequencing of Peroneutypa sp. M16 enabled the identification of a cytochalasan biosynthetic gene cluster and a proposal for the biosynthetic assembly of perochalasins. The proposal is supported by the nonenzymatic conversion of phenochalasin B (7) into 1-3, based on isotope-labeled hydroxylamine (15NH2OH and ND2OD) feeding studies in vivo and in vitro. In contrast to other merocytochalasans, these are the first cytochalasans confirmed to arise via nucleophilic addition and at a distinct location from the reactive macrocycle olefin, potentially expanding further the range of merocytochalasans to be discovered or engineered. Cytochalasin Z27 (4) exhibited antiplasmodial activities in the low micromolar range against the chloroquine-sensitive Plasmodium falciparum 3D7 strain as well as against resistant strains of the parasite (Dd2, TM90C6B, and 3D7r_MMV848).

Metabolomic Changes Associated with AGXT2 Genotype Variants and Stone Formation in a Colony of Cats.

The objective of this study was to assess serum chemistries and metabolomic parameters in cats with genetic variants of the alanine-glyoxylate aminotransferase 2 (AGXT2) gene to determine abnormalities associated with urolith formation and better understand effective approaches for the treatment of cats with uroliths. AGXT2 genotypes of 445 cats in the colony at Hill's Pet Nutrition, Inc. (Topeka, KS, USA) were assessed in a genome-wide association study. Additionally, the serum chemistries and metabolic profiles of each cat were determined, along with their lifetime history of stone incidence. Factor analysis was used as a data-reduction method for metabolites in order to perform statistical hypothesis testing and to select significant metabolites from the more than 600 serum metabolites identified. Of the 82 cats forming stones in the colony (18.4%), the majority were calcium oxalate. Results showed that approximately one third of the cats with the AA variant of the AGXT2 gene have stones, that chronic kidney disease (CKD) is more common in cats with stones, and that having stones results in a shorter lifespan. A discriminant variable selection process was performed to determine the complete blood count, serum biochemistries, and serum metabolomic factors that best discriminated among the three genotypes (AA, AG, GG) and between cats forming stones and non-stone formers. Several of the highly ranked discriminating factors included metabolites related to decreased aminotransferase activity in cats with the AA variant of the AGXT2 gene. Another factor that ranked highly for discriminating between stone formers and non-stone formers contained lipid metabolites, consisting of multiple sphingomyelin species and cholesterol. These findings support the results of feeding studies in cats, whereby CKD cats fed food supplemented with betaine and prebiotics have experienced an increase in total body mass, reduced uremic toxins, and altered sphingomyelin concentrations.

Comparative study of mash vs pellet feed on growth performance and feed utilization by goat

This study was conducted in Bharatpur-11, Chitwan, Nepal, to evaluate the effects of two feed forms (mash and pellet) on the growth performance and feed utilization in goats. Twenty Boer cross female goats, averaging five months in age, were assigned to either mash or pellet feed treatments, with 10 goats in each group, for a 90-day period, with two weeks of adaptation. The experimental animals were given concentrate feed at 1% of their body weight daily, along with ad libitum access to Napier grass and clean drinking water. Both feed types had a dry matter content of 90% and a crude protein level of 17%. Key parameters assessed included feed and forage intake, body weight changes, and feed wastage. Goats on pelleted feed showed a significantly higher average daily weight gain of 62.00 g/day compared to 26.11 g/day for those on mash feed. Final body weights were also greater in the pellet-fed group (28.88 kg) compared to the mash-fed group (25.04 kg). Additionally, pelleted feed led to higher intake of concentrate and forages and significantly reduced feed wastage, with pellet feed waste being 1.22 g/day versus 27.65 g/day for mash feed. These findings indicate that pelleted feed enhances growth performance, feed intake, and feed efficiency in goats. Therefore, this study indicated that pelleted feed is a more effective option for enhancing goat productivity and reducing feed wastage.

Preparation of umami peptides from chicken breast by ultrasound-assisted gradient dilution feeding substrate and study of their formation mechanism

Gradient dilution feeding substrate (GDFS) is regarded as an effective method for the production of active peptides, as it compensates for substrate limitations and minimizes substrate contamination of the ultrafiltration membrane. This study further investigated the impact of ultrasound-assisted gradient dilution feeding substrate (UA-GDFS) on the peptide yield and the intensity of umami. The chicken breast was used as ingredients. The umami compounds in the hydrolysate were determined using an automatic amino acid analyzer and HPLC. The hydrolysate's umami intensity was determined by electronic tongue in all modes. These findings indicated that UA-GDFS enhanced the umami intensity and peptide yield. Hydrolysate from UA-GDFS with molecular weight below 1 kDa was analyzed by nano-scale liquid chromatography-tandem mass spectrometry (Nano-HPLC-MS/MS). Electronic tongue analysis showed that GPSGPVG and VDGPSGK had umami taste and could enhance the umami taste of monosodium glutamate (MSG) solution. Asn68 and His145 were important sites on T1R3 for umami peptide binding.

PSV-29 In vitro digestibility and in situ degradability of lobster and mixed crab shell waste material

Abstract The objective of this research was to determine the in situ and in vitro degradability of lobster shell waste meal (LM) and crab shell waste meal (CM). Lobster and crab harvest are major industries within the New England states and the Maritime provinces. Harvest of the American lobster amounts to 54,643 tons caught and processed in 2020 in the New England states. The Atlantic Red Crab annual quota is around 1,814 tons. The Jonah Crab amounts to 5,725 tons. Due to the significant harvest, there is a resulting increase in byproducts and wastes at processing facilities providing potential feedstuffs for cattle. For both experiments, LM and CM were compared with soybean (SBM) and blood meals (BM). In experiment 1, 0.5 g of each meal were added to an artificial rumen incubator (Daisy, Ankom technologies) and DM degradability over 48 h was measured. In experiment 2, ruminally-cannulated Holstein cows (n = 4) in a 4 × 4 Latin square, had 5 g of sample were placed in Dacron bags in the dorsal rumen. Bags were in duplicate and removed at 2, 4, 8,12, 24, and 48 h time points. Eight bags were removed at 12 h for use in the in vitro intestinal degradability measurement using the modified three-step technique. Contrasts were: SBM vs. BM+LM+CM (SBM vs others), BM vs. LM+CM, and LM vs. CM. In experiment 2, 0 h bags served as a determination of fraction A, while the 48 h bags served as fraction C. Fraction B = 100% - (fraction A + fraction C). Degradation rate (kd) was calculated by converting % CP degraded from 2 to 24 h to Ln and then calculating the slope. In experiment 1, DM degradation was 45.8, 43.8, 76.4 and 100.1 % for SBM, BM, LM, and CM respectively; SBM vs others (P < 0.01), BM vs. LM + CM (P < 0.01), and no difference between LM vs. CM. Fraction A (CP) was 9.9, 8.1, 4.2, and 4.8% respectively; with SBM vs. others (P < 0.03), BM vs. LM+CM (P < 0.01), and no difference between LM vs CM. Fraction B (CP) was 76.2, 62.2, 28.9, and 42.6% respectively; with SBM vs. others (P < 0.01), BM vs. LM + CM (P < 0.01), and LM vs. CM (P < 0.10). Fraction C (CP) was 4.41, 30.8, 46.8, and 37.5% respectively; with SBM vs. others (P < 0.01), BM vs. LM + CM (P < 0.03), and LM vs. CM (P < 0.08). Intestinal CP digestibility was 56.2, 64.7, -5.83, and 10.5% respectively, with SBM vs others (P < 0.01), BM vs LM+CM (P < 0.01), and no difference between LM and CM. Results of these studies indicated that LM and CM may have some benefits in cattle diets, but feeding studies should be conducted.

PSIX-13 Methionine requirements across the life cycle of working Labrador Retrievers using the indicator amino acid oxidation (IAAO) technique

Abstract Methionine is a limiting amino acid in canine diets, but few requirement studies have been conducted across life stages in large-breed dogs. The objective of this study was to determine the minimum requirement (MR) for methionine (Met) in Labrador retrievers at 4 life stages using the indicator amino acid (AA) oxidation (IAAO) technique. In total, 24 dogs were used: 6 puppies aged 10 to 14 wk, 6 puppies aged 14 wk to 9 mo, 6 adults aged 2 to 5 yr, and 6 seniors aged over 8 yr in feeding studies to evaluate the changing requirements of methionine (Met) as canines age. After 2-d adaptation to Met-adequate basal diet (Met = 0.56% dry matter), dogs underwent individual IAAO studies. In brief, all dogs were randomly fed one of six test diets with varying levels of Met ranging from deficient to sufficient (final Met content in experimental diets was 0.1982, 0.2386, 0.279, 0.49, 0.525, 0.56% dry matter, with indispensable amino acids formulated at 1.6x NRC values. Test diet was divided into 13 equal meals; at the 5th meal, a tracer amino acid was supplied (a bolus L-[1-13C]phenylalanine based on body weight was first given, followed by [1-13C]Phe doses every 30 min spanning a 4-h period), and breath samples were collected via respiration mask every 30 min. Total production of 13CO2 during isotopic steady state was determined by enrichment of 13CO2 in breath samples and total CO2 production measured via indirect calorimetry and isotope ratio mass spectrometry (IRMS). Results for IRMS were converted to atom percent excess (APE) and analyzed using segmented linear regression. The Four Rivers mean and population requirements for Met were estimated to be 0.84 ± 0.05 g/1000 kcal (mean ± 2SD) for puppies 10 to 14 wk, 0.78 ± 0.16 g/1,000kcal (mean ± 2SD) for puppies 14 wk to -9 mo, 0.68 ± 0.19 g/1,000kcal (mean ± 2SD) for adults 2 to 5 yr, and 0.62 ± 0.17 g/1,000kcal (mean ± 2SD) for seniors. This research provides important information to assist in accurate formulation of diets that contain appropriate methionine for large breed dogs at all life stages.

PSLBI-17 Total sulfur amino acid requirements across the canine life cycle in Labrador Retrievers using the indicator amino acid oxidation (IAAO) technique

Abstract Methionine and cysteine are the principal sulfur-containing amino acids that are incorporated into proteins, although homocysteine and taurine also have important physiological roles in canines. As limited studies have been conducted for total sulfur-containing amino acids (TSAA) requirements across the canine life cycle, our objective was to determine the minimum requirements (MR) for TSAA at 4 life stages using the indicator amino acid oxidation (IAAO) technique. Labrador Retrievers (n = 24) were used in feeding studies to evaluate the changing requirements of TSAA as dogs mature: 6 puppies aged 10 to 14 wk, 6 puppies >14 wk to 9 mos, 6 adults 2 to 5 yr (yr), and 6 seniors > 8yr. Following a 2-d adaptation to a TSAA-adequate basal diet (TSAA = 1.12% dry matter), dogs underwent individual IAAO studies. In brief, all dogs were randomly fed one of six test diets with varying levels of TSAA ranging from deficient to sufficient (final TSAA content in experimental diets were 0.41, 0.46, 0.51, 0.98, 1.05, and 1.12% dry matter, with indispensable amino acids formulated at 1.6x NRC values). The test diet was divided into 13 equal meals; at the 5th meal, a tracer amino acid was included (a bolus L-[1-13C] phenylalanine based on body weight was first given, followed by [1-13C] Phe doses every 30 spanning 4 h). Breath samples were collected via a respiration mask every 30 min. Total CO2 production was determined via indirect calorimetry and production of 13CO2 at each dose was measured by enrichment of 13C in breath samples via isotope ratio mass spectrometry (IRMS). Results for IRMS data were converted to atom percent excess (APE) and analyzed using segmented linear regression. The Four Rivers mean and population requirements for TSAA were estimated to be 1.55 ± 0.069 g/1,000 kcal (mean ± 2 SD) for puppies 10 to 14 wk, 1.53 ± 0.21 g/1,000 kcal (mean ± 2 SD) for puppies 14 wk to 9 mo, 1.4 ± 0.30 g/1000 kcal (mean ± 2 SD) for adults 2 to 5 yr, and 1.27 ± 0.23 g/1,000 kcal (mean ± 2 SD) for seniors. These results may contribute to the formulation of diets that meet the age-appropriate requirements of TSAA for large-breed dogs at all life stages.

Effect of β-Alanine Metabolite on Gut Integrity and Immunity in Commercial Broiler Chickens Infected with Eimeria maxima.

(1) Background: In a metabolomics analysis conducted to investigate the mechanisms behind the growth-promoting effects of probiotics in broilers, β-alanine was found to be significantly elevated. This led to the hypothesis that β-alanine could also contribute to growth-promoting effects in infected broilers. (2) Methods: An in vitro culture system was developed to assess β-alanine's impact on proinflammatory cytokine response in chicken macrophage cells, gut integrity in chicken intestinal epithelial cells, and muscle differentiation in quail muscle cells and primary chicken embryonic muscle cells. In vivo animal feeding studies were then conducted to investigate the effects of dietary β-alanine on various disease parameters in Eimeria maxima-infected broiler chickens. (3) Results: In vitro, β-alanine treatment significantly decreased the gene expression of cytokines in chicken macrophage cells and increased occuldin expression in chicken intestinal epithelial cells. Dietary β-alanine increased the body weight of chickens following Eimeria maxima infection in the H-ALA group. Dietary β-alanine also suppressed cytokines and increased JAM-2 and occludin expression in the H-ALA group compared to the infected group without β-alanine supplementation. (4) Conclusions: These results strongly support the positive effects of dietary β-alanine on intestinal immune responses and gut barrier function in broiler chickens infected with Eimeria maxima.

Trajectory of Infant Problematic Feeding Symptoms: Study Protocol.

Infants with developmental risk factors are more likely to have feeding problems and develop chronic feeding disorders. Early detection and understanding of the progression of problematic feeding and its relationship with a child's biological functioning and the family feeding environment will enhance effective symptom management and development of interventions to prevent pediatric feeding disorders. The New Through Two (NewThru2) feeding study protocol is described. Study aims are to (a) characterize symptoms of problematic feeding and trajectories of symptoms from pre-discharge from neonatal intensive care through age 24 months; (b) determine the relationship of child biological function at discharge with symptom characteristics and trajectories; (c) describe the child's feeding environment and its relationship to symptoms from discharge through 24 months; and (d) determine the relationship between problematic feeding symptoms and growth and developmental outcomes. NewThru2 is a prospective, longitudinal, mixed method study following over 200 infants who received care in a neonatal intensive care unit and were identified as at risk for compromised development. The study follows enrolled infants through 24 months of age. Symptoms of problematic feeding are measured pre-discharge by clinical observation and medical record review and post-discharge by parent report. Biological function is measured by medical history and cardiorespiratory and autonomic nervous system function during feeding prior to discharge. Child feeding environment is measured by strategies parents use to manage feeding, the effect of feeding on the parent and family, and the use of feeding services. A subset of parents is interviewed to achieve a contextual understanding of the family feeding environment. Child outcome measures include parent-reported feeding skills and clinician-reported growth and neurodevelopment. The results of this study will improve understanding of pediatric feeding disorders during a time of development sensitive to adequate nutrition and with infants at risk for developmental delays or impairments.

Literature Review: Pengaruh PMT Pangan Lokal Pada Balita sebagai Upaya Pencegahan Stunting

The high prevalence of stunting is a global problem of great concern. Efforts to reduce stunting are to provide additional food to children locally to improve their nutritional status. The purpose of making this literature review is to find out the effectiveness, influence, success of the program, and efforts to provide additional food to stunted toddlers. This literature review uses several sources, namely Sinta accredited journals related to supplementary feeding which can be searched using google scholar and the SINTA database. Based on the findings of 16 studies of supplementary feeding made from local food in stunted toddlers, there was an increase in body weight and height when supplementary feeding was given for at least 30 days. Tempeh, rebon shrimp, and moringa are local foods that are easily available and relatively cheap. They also have high nutritional value. After being given tempe, rebon shrimp, and moringa, as local foods, the toddler's height increased significantly.

Rapid Screening of Colorado Potato Beetle Resistance Derived from Solanum okadae

Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say) is a major insect pest of potato and development of resistant varieties is part of a strategy for management. Wild relatives of potato are resources for genetic improvement through breeding. Interspecies crosses to introgress CPB resistance will be facilitated with rapid and inexpensive selection methods. Solanum okadae is a novel source of feeding deterrence against the beetle and was associated with production of a naturally occurring leaf-specific lactone-containing metabolite in the foliage. The Baljet assay has been used for decades in pharmaceuticals for rapid screening of lactone-containing compounds. A modified Baljet assay was developed for potato foliar tissue to rapidly screen for lactone-containing compounds in plants from the field, greenhouse, and laboratory. Herein we report the screening of potato foliage for CPB resistance with a Baljet assay validated by CPB larval feeding studies. Foliage from wild accessions of S. okadae were tested using the Baljet assay and results showed that production of the leaf-specific lactones has a large range of variation in the species. In addition, tubers tested using the Baljet assay had negligible levels of lactone-containing metabolites, confirming leaf-specific production of lactones. This inexpensive method using leaf disk screening will allow potato breeders to quickly select for potential CPB resistant germplasms and advance the breeding of sustainable crops.

Statistical approaches for component-wise censored composite endpoints.

Composite endpoints defined as the time to the earliest of two or more events are often used as primary endpoints in clinical trials. Component-wise censoring arises when different components of the composite endpoint are censored differently. We focus on a composite of death and a non-fatal event where death time is right censored and the non-fatal event time is interval censored because the event can only be detected during study visits. Such data are most often analysed using methods for right censored data, treating the time the non-fatal event was first detected as the time it occurred. This can lead to bias, particularly when the time between assessments is long. We describe several approaches for estimating the event-free survival curve and the effect of treatment on event-free survival via the hazard ratio that are specifically designed to handle component-wise censoring. We apply the methods to a randomized study of breastfeeding versus formula feeding for infants of mothers infected with human immunodeficiency virus.

An Integrated Module Performs Selective ‘On‐Line’ Epoxidation in the Biosynthesis of the Antibiotic Mupirocin.

The delineation of the complex biosynthesis of the potent antibiotic mupirocin, which consists of a mixture of pseudomonic acids (PAs) isolated from Pseudomonas fluorescens NCIMB 10586, presents significant challenges and the timing and mechanisms of several key transformations remain elusive. Particularly intriguing are the steps that process the linear backbone from the initial polyketide assembly phase to generate the first cyclic intermediate PA‐B. These include epoxidation as well as incorporation of the tetrahydropyran (THP) ring and fatty acid sidechain required for biological activity. Here, we show that the mini‐module MmpE performs a rare online (ACP‐substrate) epoxidation and is integrated (‘in‐cis’) into the polyketide synthase via a docking domain. A linear polyketide fragment with 6 asymmetric centres was synthesised using a convergent approach and used to demonstrate substrate flux via an atypical KS0 and a previously unannotated ACP (MmpE_ACP). MmpE_ACP‐bound synthetic substrates were critical in demonstrating successful epoxidation in vitro by the purified MmpE oxidoreductase domain. Alongside feeding studies, these results confirm the timing as well as chain length dependence of this selective epoxidation. These mechanistic studies pinpoint the location and nature of the polyketide substrate prior to the key formation of the THP ring and esterification that generate PA‐B.

Feeding rats with used cooking oil elevates malondialdehyde, TNF-α, and creatinine compared to tempe fried with used oil.

In vivo studies on the hazards of deep-fried foods were commonly done by feeding used-or heated-cooking oil to rats. The aim of this study was to determine the effect of feeding tempe deep-fried in palm, olive, and coconut oils and the used frying oil on the blood biochemical profile of laboratory rats. An in vivo randomized control group study with pre-test and post-test was conducted. This study included healthy male Sprague-Dawley rats aged 2-3 months and weighing 100-200 grams. After acclimatization, the rats were randomly assigned to seven groups, which were: (1) regular diet (control diet); (2) diet of tempe deep-fried in 5× used palm oil (Tempe-in-used-Po); (3) diet of tempe deep-fried in 5× used coconut oil (Tempe-in-used-Co); (4) diet of tempe deep-fried in 5× used olive oil (Tempe-in-used-Oo); (5) diet of 5× used palm oil (Used-Po); (6) diet of 5× used coconut oil (Used-Co); and (7) diet of 5× used olive oil (Used-Oo). Each rat received 15 grams of a treatment diet daily and blood samples were collected after four weeks for a complete blood count and serum biochemistry analysis. The results showed that the final body weight and the weight gain of Tempe-in-used-Po, Tempe-in-used-Co, Tempe-in-used-Oo group, and Used-Po groups increased significantly compared to the control, Used-Co, and Used-Oo groups. However, there was a significant increase in serum tumor necrosis factor-alpha (TNF-α) in the Used-Co and Used-Oo groups (p<0.05), suggesting the used oil's detrimental effect. The Used-Co and Used-Oo were the only two groups whose creatinine increased significantly (p<0.05). Subsequently, only the Used-Oo group had a significantly increased malondialdehyde (MDA) level compared to all groups (p<0.05). These results prove that the effect of feeding fried food differs from used oils. Feeding used oil did not reflect the consumption of fried foods as part of the whole diet and generally resulted in more harmful effects. This is the first study to report an in vivo rat feeding study of deep-fried tempe and the used oil as part of the diet.