Aryl amino acetamides prevent Plasmodium falciparum ring development via targeting the lipid-transfer protein PfSTART1.

With resistance to most antimalarials increasing, it is imperative that new drugs are developed. We previously identified an aryl acetamide compound, MMV006833 (M-833), that inhibited the ring-stage development of newly invaded merozoites. Here, we select parasites resistant to M-833 and identify mutations in the START lipid transfer protein (PF3D7_0104200, PfSTART1). Introducing PfSTART1 mutations into wildtype parasites reproduces resistance to M-833 as well as to more potent analogues. PfSTART1 binding to the analogues is validated using organic solvent-based Proteome Integral Solubility Alteration (Solvent PISA) assays. Imaging of invading merozoites shows the inhibitors prevent the development of ring-stage parasites potentially by inhibiting the expansion of the encasing parasitophorous vacuole membrane. The PfSTART1-targeting compounds also block transmission to mosquitoes and with multiple stages of the parasite's lifecycle being affected, PfSTART1 represents a drug target with a new mechanism of action.

Octave-spanning supercontinuum generation in a CMOS-compatible thin Si3N4 waveguide coated with highly nonlinear TeO2.

Supercontinuum generation (SCG) is an important nonlinear optical process enabling broadband light sources for many applications, for which silicon nitride (Si3N4) has emerged as a leading on-chip platform. To achieve suitable group velocity dispersion and high confinement for broadband SCG the Si3N4 waveguide layer used is typically thick (>∼700 nm), which can lead to high stress and cracks unless specialized processing steps are used. Here, we report on efficient octave-spanning SCG in a thinner moderate-confinement 400-nm Si3N4 platform using a highly nonlinear tellurium oxide (TeO2) coating. An octave supercontinuum spanning from 0.89 to 2.11 µm is achieved at a low peak power of 258 W using a 100-fs laser centered at 1565 nm. Our numerical simulations agree well with the experimental results giving a nonlinear parameter of 2.5 ± 0.5 W-1m-1, an increase by a factor of 2.5, when coating the Si3N4 waveguide with a TeO2 film. This work demonstrates highly efficient SCG via effective dispersion engineering and an enhanced nonlinearity in CMOS-compatible hybrid TeO2-Si3N4 waveguides and a promising route to monolithically integrated nonlinear, linear, and active functionalities on a single silicon photonic chip.

Initial Experiences with Amyloid-Related Imaging Abnormalities in Patients Receiving Aducanumab Following Accelerated Approval.

Aducanumab is the first FDA-approved amyloid-lowering immunotherapy for Alzheimer's disease. There is little real-world data to guide management of amyloid-related imaging abnormalities (ARIA), a potentially serious side-effect which requires surveillance with magnetic resonance imaging. We report our experiences in managing ARIA in patients receiving aducanumab at the Butler Hospital Memory and Aging Program during the year following FDA approval. We followed the Appropriate Use Recommendations for aducanumab to guide patient selection, detection, and management of ARIA (1). ARIA-E occurred in 6 out of 24 participants treated; all APOE-ε4 carriers. Treatment was discontinued in 4 cases of moderate-severe ARIA-E, temporarily held in 1 moderate case, and dosed through in 1 mild case (mean duration = 3 months, range, 1-6 months). No participants required hospitalization or high dose corticosteroids. Participants on anticoagulation were excluded and no macrohemorrhages occurred. These data support the measured approaches to treatment outlined in the Appropriate Use Recommendations.

Visual access to an outdoor range early in life, but not environmental complexity, increases meat chicken ranging behavior.

Not all chickens access an outdoor range when the opportunity is provided. This may be related to the abrupt change in environments from the stable rearing conditions to the complexity of the outdoor range. We aimed to prepare chickens to range by increasing the complexity of the indoor environment early in life with the intention to encourage range use. Mixed sex Cobb500 chickens were allocated to 1 of 3 treatment groups: visual access (VA) treatment provided VA to the outdoor range from day old via transparent pop-hole covers; environmental complexity (EC) treatment provided an artificial haybale, fan with streamers and a solid vertical barrier; Control treatment was a representative conventional environment. Chickens were given access to the outdoor range at 21 d of age. Behavior in the home pen was assessed in wk 1, 2 and 5 and individual ranging behavior was monitored through radio frequency identification (RFID) technology. The VA chickens were more active compared to EC (P = 0.006) and Control (P = 0.007) chickens and spent more time foraging than control chickens (P = 0.036) during the first week of life. More VA chickens accessed the range area compared to EC chickens (P = 0.015). VA chickens accessed the range sooner after they were first provided access and spent more time on the range than EC and control chickens (P < 0.001). Mortality was lower in the VA treatment compared to EC (P = 0.024) and control group (P = 0.002). There was evidence that VA chickens weighed less than Control and EC chickens, however results were inconsistent between age and sex. Hence, providing meat chickens with VA to an outdoor range early in life increased activity in early life, decreased latency to first access the range and increased time on the range and lowered mortality. Future work should aim to understand the mechanism behind these changes in behavior to develop recommendations for producers to implement in commercial conditions.

Dietary Flaxseed and Flaxseed Oil Differentially Modulate Aspects of the Microbiota Gut-Brain Axis Following an Acute Lipopolysaccharide Challenge in Male C57Bl/6 Mice.

The microbiota gut-brain axis (mGBA) is an important contributor to mental health and neurological and mood disorders. Lipopolysaccharides (LPS) are endotoxins that are components of Gram-negative bacteria cell walls and have been widely shown to induce both systemic and neuro-inflammation. Flaxseed (Linum usitatissimum) is an oilseed rich in fibre, n3-poly-unsaturated fatty acid (alpha-linolenic acid (ALA)), and lignan, secoisolariciresinol diglucoside, which all can induce beneficial effects across varying aspects of the mGBA. The objective of this study was to determine the potential for dietary supplementation with flaxseed or flaxseed oil to attenuate LPS-induced inflammation through modulation of the mGBA. In this study, 72 5-week-old male C57Bl/6 mice were fed one of three isocaloric diets for 3 weeks: (1) AIN-93G basal diet (BD), (2) BD + 10% flaxseed (FS), or (3) BD + 4% FS oil (FO). Mice were then injected with LPS (1 mg/kg i.p) or saline (n = 12/group) and samples were collected 24 h post-injection. Dietary supplementation with FS, but not FO, partially attenuated LPS-induced systemic (serum TNF-α and IL-10) and neuro-inflammation (hippocampal and/or medial prefrontal cortex IL-10, TNF-α, IL-1ß mRNA expression), but had no effect on sickness and nest-building behaviours. FS-fed mice had enhanced fecal microbial diversity with increased relative abundance of beneficial microbial groups (i.e., Lachnospiraceae, Bifidobacterium, Coriobacteriaceae), reduced Akkermansia muciniphila, and increased production of short-chain fatty acids (SCFAs), which may play a role in its anti-inflammatory response. Overall, this study highlights the potential for flaxseed to attenuate LPS-induced inflammation, in part through modulation of the intestinal microbiota, an effect which may not be solely driven by its ALA-rich oil component.

A Pyridyl-Furan Series Developed from the Open Global Health Library Block Red Blood Cell Invasion and Protein Trafficking in Plasmodium falciparum through Potential Inhibition of the Parasite's PI4KIIIB Enzyme.

With the resistance increasing to current antimalarial medicines, there is an urgent need to discover new drug targets and to develop new medicines against these targets. We therefore screened the Open Global Health Library of Merck KGaA, Darmstadt, Germany, of 250 compounds against the asexual blood stage of the deadliest malarial parasite Plasmodium falciparum, from which eight inhibitors with low micromolar potency were found. Due to its combined potencies against parasite growth and inhibition of red blood cell invasion, the pyridyl-furan compound OGHL250 was prioritized for further optimization. The potency of the series lead compound (WEHI-518) was improved 250-fold to low nanomolar levels against parasite blood-stage growth. Parasites selected for resistance to a related compound, MMV396797, were also resistant to WEHI-518 as well as KDU731, an inhibitor of the phosphatidylinositol kinase PfPI4KIIIB, suggesting that this kinase is the target of the pyridyl-furan series. Inhibition of PfPI4KIIIB blocks multiple stages of the parasite's life cycle and other potent inhibitors are currently under preclinical development. MMV396797-resistant parasites possess an E1316D mutation in PfPKI4IIIB that clusters with known resistance mutations of other inhibitors of the kinase. Building upon earlier studies that showed that PfPI4KIIIB inhibitors block the development of the invasive merozoite parasite stage, we show that members of the pyridyl-furan series also block invasion and/or the conversion of merozoites into ring-stage intracellular parasites through inhibition of protein secretion and export into red blood cells.

Hybrid silicon-tellurium-dioxide DBR resonators coated in PMMA for biological sensing.

We report on silicon waveguide distributed Bragg reflector (DBR) cavities hybridized with a tellurium dioxide (TeO2) cladding and coated in plasma functionalized poly (methyl methacrylate) (PMMA) for label free biological sensors. We describe the device structure and fabrication steps, including reactive sputtering of TeO2 and spin coating and plasma functionalization of PMMA on foundry processed Si chips, as well as the characterization of two DBR designs via thermal, water, and bovine serum albumin (BSA) protein sensing. Plasma treatment on the PMMA films was shown to decrease the water droplet contact angle from ∼70 to ∼35°, increasing hydrophilicity for liquid sensing, while adding functional groups on the surface of the sensors intended to assist with immobilization of BSA molecules. Thermal, water and protein sensing were demonstrated on two DBR designs, including waveguide-connected sidewall (SW) and waveguide-adjacent multi-piece (MP) gratings. Limits of detection of 60 and 300 × 10-4 RIU were measured via water sensing, and thermal sensitivities of 0.11 and 0.13 nm/°C were measured from 25-50 °C for SW and MP DBR cavities, respectively. Plasma treatment was shown to enable protein immobilization and sensing of BSA molecules at a concentration of 2 µg/mL diluted in phosphate buffered saline, demonstrating a ∼1.6 nm resonance shift and subsequent full recovery to baseline after stripping the proteins with sodium dodecyl sulfate for a MP DBR device. These results are a promising step towards active and laser-based sensors using rare-earth-doped TeO2 in silicon photonic circuits, which can be subsequently coated in PMMA and functionalized via plasma treatment for label free biological sensing.

Erbium-ytterbium co-doped aluminium oxide waveguide amplifiers fabricated by reactive co-sputtering and wet chemical etching.

We report on the fabrication and optical characterization of erbium-ytterbium co-doped aluminum oxide (Al2O3:Er3+:Yb3+) waveguides using low-cost, low-temperature deposition and etching steps. We deposited Al2O3:Er3+:Yb3+ films using reactive co-sputtering, with Er3+ and Yb3+ ion concentrations ranging from 1.4-1.6 × 1020 and 0.9-2.1 × 1020 ions/cm3, respectively. We etched ridge waveguides in 85% pure phosphoric acid at 60°C, allowing for structures with minimal polarization sensitivity and acceptable bend radius suitable for optical amplifiers and avoiding alternative etching chemistries which use hazardous gases. Scanning-electron-microscopy (SEM) and profilometry were used to assess the etch depth, sidewall roughness, and facet profile of the waveguides. The Al2O3:Er3+:Yb3+ films exhibit a background loss as low as 0.2 ± 0.1 dB/cm and the waveguide loss after structuring is determined to be 0.5 ± 0.3 dB/cm at 1640 nm. Internal net gain of 4.3 ± 0.9 dB is demonstrated at 1533 nm for a 3.0 cm long waveguide when pumped at 970 nm. The material system is promising moving forward for compact Er-Yb co-doped waveguide amplifiers and lasers on a low-cost silicon wafer-scale platform.

Leukocyte-dependent effects of platelet-rich plasma on cartilage loss and thermal hyperalgesia in a mouse model of post-traumatic osteoarthritis.

OBJECTIVE: Platelet-rich plasma (PRP) is an emerging therapeutic strategy for treatment of osteoarthritis (OA); however, there is a lack of preclinical and clinical evidence for its efficacy and its mechanism of action is unclear. In the current study, we utilized leukocyte poor-PRP (LP-PRP) and leukocyte rich-PRP (LR-PRP) to mimic clinical point of care formulations and assessed their potential to alter disease progression in a mouse model of post-traumatic OA. METHOD: Three-month-old wild-type male FVB/N mice received destabilization of the medial meniscus (DMM) surgery to induce OA. To assess the efficacy of LP-PRP and LR-PRP, mice were given intraarticular injections at 2-, 7- and 28-days post-surgery. Mice were then assessed at 5-, 9-, and 13-weeks post-surgery for changes in chronic pain using the hot plate nociceptive assay. At 14-weeks, OA pathogenesis was evaluated using histology and phase-contrast µCT. RESULTS: Treatment with LP-PRP and to a lesser extent LR-PRP preserved cartilage volume and surface area compared to phosphate-buffered saline (PBS) as measured by phase-contrast µCT. However, both treatments had higher Osteoarthritis Research Society International (OARSI) and synovitis scores compared to sham, and neither substantially improved scores compared to PBS controls. With respect to thermal hyperalgesia, PBS-treated mice displayed reduced latency to response compared to sham, and LR-PRP but not LP-PRP improved latency to response at 5-, 9- and 13-weeks post-surgery compared to PBS. CONCLUSION: The results of this study suggest that effects of PRP therapy on OA progression and disease-induced hyperalgesia may be leukocyte-dependent. And while LP-PRP and to a lesser extent LR-PRP protect from volume and surface loss, significant pathology is still seen within OA joints. Future work is needed to understand how the different components of PRP effect OA pathogenesis and pain, and how these could be modified to achieve greater therapeutic efficacy.

Active transport of brilliant blue FCF across the Drosophila midgut and Malpighian tubule epithelia.

Under conditions of stress, many animals suffer from epithelial barrier disruption that can cause molecules to leak down their concentration gradients, potentially causing a loss of organismal homeostasis, further injury or death. Drosophila is a common insect model, used to study barrier disruption related to aging, traumatic injury, or environmental stress. Net leak of a non-toxic dye (Brilliant blue FCF) from the gut lumen to the hemolymph is often used to identify barrier failure under these conditions, but Drosophila are capable of actively transporting structurally-similar compounds. Here, we examined whether cold stress (like other stresses) causes Brilliant blue FCF (BB-FCF) to appear in the hemolymph of flies fed the dye, and if so whether Drosophila are capable of clearing this dye from their body following chilling. Using in situ midgut leak and transport assays as well as Ramsay assays of Malpighian tubule transport, we tested whether these ionoregulatory epithelia can actively transport BB-FCF. In doing so, we found that the Drosophila midgut and Malpighian tubules can mobilize BB-FCF via an active transcellular pathway, suggesting that elevated concentrations of the dye in the hemolymph may occur from increased paracellular permeability, reduced transcellular clearance, or both. SUMMARY STATEMENT: Drosophila are able to actively secrete Brilliant blue FCF, a commonly used marker of barrier dysfunction.

Flock use of the range is associated with the use of different components of a multi-tier aviary system in commercial free-range laying hens.

1. The objective of this study was to investigate the association of using a multi-tier aviary system and access to range on flock uniformity in free-range laying hens, and to determine whether the extent of range use or flock uniformity can be predicted from the use of different levels of the aviary system.2. A total of 13,716 Lohmann Brown hens from five commercial free-range flocks housed in identical houses on the same farm were individually weighed at 16 weeks of age and allocated to five replicate areas within each house. Hen movement in the multi-tier aviary system and on the range was individually monitored using radio frequency identification (RFID). All hens had access to the range from 18 to 22 weeks of age and were exposed to the same management conditions.3. Whilst only one flock significantly changed its flock uniformity with time, they differed from each other in uniformity and body weight (P = 0.001).4. Hens spent most of their available time on the lower aviary tier (7.29 ± 0.029 h/hen/day) and on the upper aviary tier (4.29 ± 0.024 h/hen/day) while the least amount of time was spent on the range and in the nest boxes (0.93 ± 0.005 h/hen/day and 1.48 ± 0.007 h, respectively, P = 0.001).5. Range use was negatively correlated (r = -0.30) to the time spent on the upper aviary tier and positively correlated (r = 0.46) to the time spent on the lower aviary tier (P = 0.001). Bivariate analysis revealed that range and upper aviary resp. lower aviary tier usage had a significant curvilinear association.6. In conclusion, the study showed that range use was associated to the time hens spent on the different tiers of the aviary system. Flock uniformity varied between flocks but was not associated with either range and aviary system usage.

Low-loss TeO2-coated Si3N4 waveguides for application in photonic integrated circuits.

We report on high-quality tellurium oxide waveguides integrated on a low-loss silicon nitride wafer-scale platform. The waveguides consist of silicon nitride strip features, which are fabricated using a standard foundry process and a tellurium oxide coating layer that is deposited in a single post-processing step. We show that by adjusting the Si3N4 strip height and width and TeO2 layer thickness, a small mode area, small bend radius and high optical intensity overlap with the TeO2 can be obtained. We investigate transmission at 635, 980, 1310, 1550 and 2000 nm wavelengths in paperclip waveguide structures and obtain low propagation losses down to 0.6 dB/cm at 2000 nm. These results illustrate the potential for compact linear, nonlinear and active tellurite glass devices in silicon nitride photonic integrated circuits operating from the visible to mid-infrared.

A Tellurium Oxide Microcavity Resonator Sensor Integrated On-Chip with a Silicon Waveguide.

We report on thermal and evanescent field sensing from a tellurium oxide optical microcavity resonator on a silicon photonics platform. The on-chip resonator structure is fabricated using silicon-photonics-compatible processing steps and consists of a silicon-on-insulator waveguide next to a circular trench that is coated in a tellurium oxide film. We characterize the device's sensitivity by both changing the temperature and coating water over the chip and measuring the corresponding shift in the cavity resonance wavelength for different tellurium oxide film thicknesses. We obtain a thermal sensitivity of up to 47 pm/°C and a limit of detection of 2.2 × 10-3 RIU for a device with an evanescent field sensitivity of 10.6 nm/RIU. These results demonstrate a promising approach to integrating tellurium oxide and other novel microcavity materials into silicon microphotonic circuits for new sensing applications.

The need for an alternative method to determine intravascular volumes.

It is well described that numerous environmental factors, including exercise, modulate plasma volume (PV). These modulations prove problematic when a number of haematological markers are measured as a concentration in blood plasma. A primary example is haemoglobin concentration ([Hb]), a marker of erythropoiesis commonly used within medicine and also used to detect blood doping. Natural changes in PV can confound [Hb] values when a volume change is detected rather than a true change in haemoglobin mass (Hbmass) (e.g. volume expansion resulting in a [Hb] decrease and pseudo-anemia vs. Hbmass decline resulting in anaemia). Currently, there is no simple solution to correct for PV shifts, and this has proven problematic when monitoring volumetric health markers in clinical and anti-doping settings. This narrative review explores the influence that PV shifts have on volumetric biomarkers, such as [Hb]. The progressive expansion in PV observed during multi-day endurance events will be summarised, and the observed impact PV variance has on concentration-based markers will be quantified. From this, the need for alternative methods to correct [Hb] for volume fluctuations is highlighted. Available methods for calculating intravascular volumes are then discussed, with a focus on a recently developed approach using a panel of 'volume descriptive' biomarkers from a standard blood test. Finally, the practical applications of this novel PV blood test within both anti-doping and clinical settings will be examined.

The effects of nasal lavage with betamethasone cream post-endoscopic sinus surgery: clinical trial.

BACKGROUND: Steroid nasal irrigation for chronic rhinosinusitis patients following endoscopic sinus surgery reduces symptom recurrence. There are minimal safety data to recommend this treatment. This study evaluated the safety of betamethasone nasal irrigation by measuring its impact on endogenous cortisol levels. METHODS: Participants performed daily betamethasone nasal irrigation for six weeks. The impact on pre- and post-intervention serum and 24-hour urinary free cortisol was assessed. Efficacy was evaluated using the 22-item Sino-Nasal Outcome Test. RESULTS: Thirty participants completed the study (16 females and 14 males; mean age = 53.9 ± 15.6 years). Serum cortisol levels were unchanged (p = 0.28). However, 24-hour urinary free cortisol levels decreased (47.5 vs 41.5 nmol per 24 hours; p = 0.025). Sino-Nasal Outcome Test scores improved (41.13 ± 21.94 vs 23.4 ± 18.17; p < 0.001). The minimal clinical important difference was reached in 63 per cent of participants. CONCLUSION: Daily betamethasone nasal irrigation is an efficacious treatment modality not associated with changes in morning serum cortisol levels. The changes in 24-hour urinary free cortisol levels are considered clinically negligible. Hence, continued use of betamethasone nasal irrigation remains a viable and safe treatment option for chronic rhinosinusitis patients following functional endoscopic sinus surgery.

Lubricity of gold nanocrystals on graphene measured using quartz crystal microbalance.

In order to test recently predicted ballistic nanofriction (ultra-low drag and enhanced lubricity) of gold nanocrystals on graphite at high surface speeds, we use the quartz microbalance technique to measure the impact of deposition of gold nanocrystals on graphene. We analyze our measurements of changes in frequency and dissipation induced by nanocrystals using a framework developed for friction of adatoms on various surfaces. We find the lubricity of gold nanocrystals on graphene to be even higher than that predicted for the ballistic nanofriction, confirming the enhanced lubricity predicted at high surface speeds. Our complementary molecular dynamics simulations indicate that such high lubricity is due to the interaction strength between gold nanocrystals and graphene being lower than previously assumed for gold nanocrystals and graphite.

Formal evaluation of PYNK: Breast Cancer Program for Young Women-the patient perspective.

PURPOSE: The aim of the present study was to assess patient satisfaction with pynk: Breast Cancer Program for Young Women so as to determine how the program might be improved and to provide feedback to donors. METHODS: All pynk patients who had consented to have their information entered in our database and who supplied us with their e-mail address were invited to complete a 58-item online questionnaire consisting of multiple choice and open-ended questions. Domains included demographics, provision of written and spoken information, support, infertility risk, research awareness, attitudes toward discharge, and general feedback. RESULTS: Of 120 pynk patients approached, 61 (51%) participated. More than 90% were satisfied or very satisfied with the timing, usefulness, and clarity of spoken and written information given, and 69% found the service and support provided by the nurse navigator to be the most helpful component of the program. Of those who had received systemic therapy, 93% recalled a health care provider initiating a discussion of the risk of treatment-related infertility, and 67% were referred to a fertility clinic. On the negative side, 11%-27% were unaware of various services provided by pynk, and 11% were unaware of pynk's ongoing research. One third of patients were unhappy or ambivalent about the prospect of discharge from the program. CONCLUSIONS: Patient satisfaction with this novel program for young women with breast cancer is high. This study highlights the critical role that the nurse navigator plays in patient support and dissemination of information. In contrast to other reported surveys of young cancer patients, pynk patients are routinely given the opportunity to undergo fertility preservation.

Neurocognitive Effects Following Cranial Irradiation for Brain Metastases.

About 90% of patients with brain metastases have impaired neurocognitive function at diagnosis and up to two-thirds will show further declines within 2-6 months of whole brain radiotherapy. Distinguishing treatment effects from progressive disease can be challenging because the prognosis remains poor in many patients. Omitting whole brain radiotherapy after local therapy in good prognosis patients improves verbal memory at 4 months, but the effect of higher intracranial recurrence and salvage therapy rates on neurocognitive function beyond this time point is unknown. Hippocampal-sparing whole brain radiotherapy and postoperative stereotactic radiosurgery are investigational techniques intended to reduce toxicity. Here we describe the changes that can occur and review technological, pharmacological and practical approaches used to mitigate their effect in clinical practice.

Effect of sodium phosphate supplementation on repeated high-intensity cycling efforts.

Limited research has investigated how sodium phosphate supplementation affects exercise performance typical of athletic competition and whether any effects linger in the short term. This study examined the effect of sodium phosphate supplementation on a cycling protocol consisting of repeated-sprint (4 sets of 6 × 15 s) and time-trial (2 × 5 min) efforts on day 1 and 4 post-loading. Trained male cyclists (VO(2peak) 5.3 L · min⁻¹) were randomised to 6 days of sodium phosphate supplementation (50 mg · kg·fat-free-mass⁻¹ · day⁻¹; n = 7) or placebo (n = 10). Performance was assessed at baseline and 1 and 4 days post-supplementation on an air-braked cycle ergometer. Compared with baseline, the sodium phosphate group recorded significantly improved (P < 0.05) work and mean power output values in both the sprint (baseline, 259 kJ/719 W; day 1, 271 kJ/754 W; day 4, 271 kJ/753 W) and time-trial (baseline, 225 kJ/374 W; day 1, 235 kJ/398 W; day 4, 236 kJ/393 W) aspects of the performance test post-loading. In the placebo group, no differences (P > 0.05) in total work or power output were noted in response to supplementation. In summary, sodium phosphate supplementation improved repeated-sprint and time-trial cycling efforts both 1 and 4 days post-loading in trained cyclists.

Argininosuccinate lyase in enterocytes protects from development of necrotizing enterocolitis.

Necrotizing enterocolitis (NEC), the most common neonatal gastrointestinal emergency, results in significant mortality and morbidity, yet its pathogenesis remains unclear. Argininosuccinate lyase (ASL) is the only enzyme in mammals that is capable of synthesizing arginine. Arginine has several homeostatic roles in the gut and its deficiency has been associated with NEC. Because enterocytes are the primary sites of arginine synthesis in neonatal mammals, we evaluated the consequences of disruption of arginine synthesis in the enterocytes on the pathogenesis of NEC. We devised a novel approach to study the role of enterocyte-derived ASL in NEC by generating and characterizing a mouse model with enterocyte-specific deletion of Asl (Asl(flox/flox); VillinCre(tg/+), or CKO). We hypothesized that the presence of ASL in a cell-specific manner in the enterocytes is protective in the pathogenesis of NEC. Loss of ASL in enterocytes resulted in an increased incidence of NEC that was associated with a proinflammatory state and increased enterocyte apoptosis. Knockdown of ASL in intestinal epithelial cell lines resulted in decreased migration in response to lipopolysaccharide. Our results show that enterocyte-derived ASL has a protective role in NEC.