Medium Chain Triglyceride Oil Research Articles

3D printed MCT oleogel as a co-delivery carrier for curcumin and resveratrol

Designing a suitable matrix to protect sensitive bioactive compounds is an important stage in nutraceutical development. In this study, emulsion templated medium-chain triglycerides (MCT) oleogel was developed as co-delivery carriers for synergistic nutraceuticals, curcumin, and resveratrol and to 3D print in customized shapes for personalized nutrition. To obtain the stable emulsion, gelatin and gellan gum were added such that their protein-polysaccharide interaction helps in the structuring of the oil phase. Increasing the amount of gellan gum had a positive effect on stabilizing the emulsion but became the critical parameter during 3D printing. Hence, gellan gum of 1.5% (w/v) and gelatin at 10% (w/v) of water were considered optimum to produce a stable 30% O/W emulsion for 3D printing. Upon analyzing the in-vitro digestion behavior of the oleogel, it was observed that the bioactives were protected under oral and gastric conditions and allowed intestinal targeted delivery. The total bioaccessible fraction increased up to 1.13-fold and 1.2-fold for curcumin and resveratrol respectively compared to control (MCT oil). The FFAs release profile also indicated that gelators play an important role in lipase activity. Also, the ex-vivo everted gut sac analysis showed enhanced permeation of about 1.83 times and 1.13 times for curcumin and resveratrol respectively. Thus, this study provides useful insights into the 3D printing of emulsion templated oleogel as personalized nutraceutical carriers.

Minimal Effects of Medium-Chain Triglyceride Supplementation on the Intestinal Microbiome Composition of Premature Infants: A Single-Center Pilot Study.

Compared to term infants, the microbiota of preterm infants is less diverse and often enriched for potential pathogens (e.g., members of the family Enterobacteriaceae). Additionally, antibiotics are frequently given to preterm infants, further destabilizing the microbiota and increasing the risk of fungal infections. In a previous communication, our group showed that supplementation of the premature infant diet with medium-chain triglyceride (MCT) oil reduced the fungal burden of Candida spp. in the gastrointestinal tract. The objective of this study was to determine whether MCT supplementation impacts the bacterial component of the microbiome. Pre-term infants (n = 17) receiving enteral feedings of either infant formula (n = 12) or human milk (n = 5) were randomized to MCT supplementation (n = 9) or no supplementation (n = 8). Fecal samples were taken at randomization and prior to MCT supplementation (Week 0), on days 5–7 (Week 1) and day 21 (Week 3). After DNA extraction from samples, the QIIME2 pipeline was utilized to measure community diversity and composition (genera and phyla). Our findings show that MCT supplementation did not significantly alter microbiota diversity or composition in the gastrointestinal tract. Importantly, there were no significant changes in the family Enterobacteriaceae, suggesting that MCT supplementation did not enrich for potential pathogens. MCT holds promise as a therapeutic intervention for reducing fungal colonization without significant impact on the bacterial composition of the host gastrointestinal tract.

Efficacy of supplemental MCT oil on seizure reduction of adult drug-resistant epilepsy – a single-center open-label pilot study

ABSTRACT Objective We set out to determine whether adding medium chain triglyceride (MCT) oil as a dietary supplement to standard diet in adult subjects with intractable epilepsy in a U.S. neurology clinical practice was associated with a reduction in number of seizures. We secondarily aimed to determine whether subjects experienced any side effects and whether there was a presence of urinary ketones while using MCT oil as a dietary supplement. Methods Adult patients with intractable epilepsy were recruited at standard of care clinical visits with their epileptologist. Once enrolled, subjects were instructed to supplement their diet with MCT oil as tolerated twice daily for three months (including a 1–2 week titration period, followed by a 1–2 week tapering off window) while keeping a seizure diary to record total number of seizures, presence of urinary ketones, and any side effects. Results Our data although limited by small sample size, shows that there is an estimated 42% reduction (p < 0.0001) in the rate of seizures. The MCT oil supplementation was well tolerated by most subjects except for minor GI side effects like nausea and loose stools. Most subjects developed ketones in their urine at some point during the trial. Conclusions MCT oil supplementation reduced seizure frequency in study participants. The reported side effects included mild nausea, stomachache, loose stools. A placebo-controlled study will be more informative.

Oil-Based Delivery Control Release System Targeted to the Later Part of the Gastrointestinal Tract-A Mechanistic Study.

Oil-based drug delivery systems have been studied in different aspects. The present study proposes a new application for an oil-based delivery system, focusing on controlled release until the drug reaches the later part of the small intestine. Bulk surfactants and interfacial surfactants were added into the oil formulation to provide a better mechanistic understating of the lipolysis. Validation of the modified in vitro method shows the overall conversion from medium-chain triglyceride oil (MCT oil) to free fatty acids (FFA) of 100 ± 4% in five replicates. This fully converted level and high reproducibility are fundamental for the following investigations where any retarding effect can be distinguished from the experimental errors. The results show that viscosity and thermodynamic activity have limited retardation. Furthermore, the former may change the kinetics of lipolysis, while the latter changes the equilibrium level. The gel-forming retarder (ethylcellulose) displayed a strong effect. Whereas the lipolysis was significantly retarded (>50%) when the retarders altered the interfacial composition (poloxamer 407), degradable interfacial surfactants did not have the same effect. However, surface-active, lipolysis-resistant retarders with a high CMC did not show a retarding effect.

Antiseizure Effects of Cannabidiol Leading to Increased Peroxisome Proliferator-Activated Receptor Gamma Levels in the Hippocampal CA3 Subfield of Epileptic Rats.

We evaluated the effects of cannabidiol (CBD) on seizures and peroxisome proliferator-activated receptor gamma (PPARγ) levels in an animal model of temporal lobe epilepsy (TLE). Adult male Sprague-Dawley rats were continuously monitored by video-electrocorticography up to 10 weeks after an intraperitoneal kainic acid (15 mg/kg) injection. Sixty-seven days after the induction of status epilepticus and the appearance of spontaneous recurrent seizures in all rats, CBD was dissolved in medium-chain triglyceride (MCT) oil and administered subcutaneously at 120 mg/kg (n = 10) or 12 mg/kg (n = 10), twice a day for three days. Similarly, the vehicle was administered to ten epileptic rats. Brain levels of PPARγ immunoreactivity were compared to those of six healthy controls. CBD at 120 mg/kg abolished the seizures in 50% of rats (p = 0.033 vs. pre-treatment, Fisher’s exact test) and reduced total seizure duration (p < 0.05, Tukey Test) and occurrence (p < 0.05). PPARγ levels increased with CBD in the hippocampal CA1 subfield and subiculum (p < 0.05 vs. controls, Holm–Šidák test), but only the highest dose increased the immunoreactivity in the hippocampal CA3 subfield (p < 0.001), perirhinal cortex, and amygdala (p < 0.05). Overall, these results suggest that the antiseizure effects of CBD are associated with upregulation of PPARγ in the hippocampal CA3 region.

Examining the Effect of Consuming C8 Medium-Chain Triglyceride Oil for 14 Days on Markers of NLRP3 Activation in Healthy Humans

Chronic, low-grade inflammation is associated with the development of numerous diseases and is mediated in part by overactivation of the NLRP3 inflammasome. The ketone body beta-hydroxybutyrate (βHB) suppresses the NLRP3 inflammasome and alters intracellular signalling pathways in vitro and in animal models; however, this effect has not yet been shown in vivo in humans. The purpose of this single-arm pilot trial was to determine if consuming 15 mL of C8 medium-chain triglyceride (trioctanoin; MCT) oil, which induces mild elevation of βHB, twice daily (30 mL total) for 14 days would suppress markers of NLRP3 inflammasome activation in young, healthy humans while following their habitual diet. Consuming a single dose of 15 mL of C8 MCT oil significantly raised blood βHB from fasting at 60 minutes and 120 minutes post ingestion (both P < 0.05). However, consumption of C8 MCT oil for 14 days did not impact markers of monocyte NLRP3 inflammasome activation compared to baseline. Specifically, caspase-1 activation and secretion of its downstream product interleukin (IL)-1β were unchanged following 14 days of C8 MCT oil supplementation when measured in unstimulated and LPS-stimulated whole blood cultures (all P > 0.05). Acetylation of histone H3 on the lysine residue 9 was unchanged (P < 0.05) and acetylation of lysine residue 14 was decreased (P < 0.05) following 14 days of supplementation. Thus, adding twice daily C8 MCT oil supplementation to the habitual diet of young, healthy humans does not appear to suppress NLRP3 inflammasome activation.

Impact of medium-chain triglycerides on gait performance and brain metabolic network in healthy older adults: a double-blind, randomized controlled study.

Nutritional supplementation with medium-chain triglycerides (MCTs) has the potential to increase memory function in elderly patients with frailty and dementia. Our aim was to investigate the effects of MCT on cognitive and gait functions and their relationships with focal brain metabolism and functional connectivity even in healthy older adults. Participants were blindly randomized and allocated to two groups: 18g/day of MCT oil and matching placebo formula (control) administered as a jelly stick (6g/pack, ingested three times a day). Gait analysis during the 6-m walk test, cognition, brain focal glucose metabolism quantified by 18F-fluorodeocyglucose positron emission tomography, and magnetic resonance imaging-based functional connectivity were assessed before and after a 3-month intervention. Sixty-three healthy, normal adults (females and males) were included. Compared with the control group, the MCT group showed better balance ability, as represented by the lower Lissajous index (23.1 ± 14.4 vs. 31.3 ± 18.9; P < 0.01), although no time × group interaction was observed in cognitive and other gait parameters. Moreover, MCT led to suppressed glucose metabolism in the right sensorimotor cortex compared with the control (P < 0.001), which was related to improved balance (r = 0.37; P = 0.04) along with increased functional connectivity from the ipsilateral cerebellar hemisphere. In conclusion, a 3-month MCT supplementation improves walking balance by suppressing glucose metabolism, which suggests the involvement of the cerebro-cerebellar network. This may reflect, at least in part, the inverse reaction of the ketogenic switch as a beneficial effect of long-term MCT dietary treatment.

Stability Testing of Sirolimus in Medium-Chain Triglyceride Oil Using High-Performance Liquid Chromatography.

Recently, a premature neonate, born at 26.5 weeks, was treated with sirolimus for kaposiform hemangioendothelioma with Kasabach-Merritt phenomenon at the University of New Mexico Children's Hospital. Because of the lipophilic properties of the drug and an inability to draw the correct dose needed for the neonate with the standard 1 mg/mL concentration, sirolimus was diluted to 0.01 mg/mL (10 mg/L) in medium-chain triglyceride (MCT) oil. The objective of this study was to evaluate the stability of sirolimus diluted in MCT oil. Commercially available sirolimus oral solution was diluted with MCT oil from 1 mg/mL to 10 mg/L. The diluted samples were prepared by measuring 0.1 mL of the commercial product in an oral syringe and mixing with 9.9 mL of MCT oil. The 3 diluted samples were placed in amber glass vials, stored at 10°C, and analyzed over 14 days. The initial concentration of the 3 samples ranged from 8.5 to 10.8 mg/L sirolimus, or 85% to 108% of the target value. No significant differences were seen between average concentrations on the days tested. However, the average drug concentration fell to approximately 90% of the theoretical 10 mg/L target concentration between days 7 and 10. This study shows that sirolimus oral solution diluted with MCT oil to a final concentration of 10 mg/L was stable for at least 7 days when stored at 10°C, suggesting that the beyond-use date should be 7 days.

Improving curcumin retention in oil-in-water emulsions coated by chitosan and their disperse stability exposed to thermal treatments

Curcumin-loaded (6% medium-chain triglyceride oil in water) emulsions were coated with chitosan and then treated with different thermal processes, including LTLT (low-temperature, long-time), HTST (high-temperature, short-time), and autoclave treatments. The mean size and zeta-potential of the emulsions linearly increased with increasing chitosan concentrations. All emulsions maintained their initial mean size after LTLT and HTST, but autoclaving increased the emulsion sizes. The size increment diminished with increasing chitosan concentrations. Moreover, 26.1% of the curcumin in non-coated emulsions was degraded during the autoclaving, whereas 83.1% of the curcumin was retained in emulsions coated with 1.0% chitosan. Thus, the emulsions coated with 1% chitosan are applicable to various thermal processing including LTLT, HTST, and autoclaving. All emulsions showed Newtonian fluid behavior, but viscosity increased with increasing chitosan concentrations. Therefore, the emulsion coated with a high chitosan concentration (more than 0.5%) showed enhanced storage stability in turbidimetric analysis despite its high polydispersity index. • Curcumin loaded emulsions were coated with different concentrations of chitosan. • Curcumin retention in emulsions was determined after thermal processing. • Chitosan coating protected the curcumin encapsulated in emulsions against heat. • Chitosan coated emulsions exhibited Newtonian fluid behavior. • Chitosan coating prevented creaming formation of emulsions by destabilization.

CHARACTERISTICS AND STABILITY OF IBUPROFEN MICROEMULSION PER INJECTION USING A COMBINATION OF MEDIUM-CHAIN TRIGLYCERIDE OIL, EXTRA VIRGIN OLIVE OIL, AND VIRGIN COCONUT OIL AS THE OIL PHASE

Injectable ibuprofen has been formulated as nanoemulsion, prodrug microemulsion, and freeze-dried. However, its application is limited because of solubility, high dosing, and complex formulation. Microemulsion ibuprofen injection is an alternative dosage form that increases solubility and bioavailability. The study aimed to formulate an ibuprofen microemulsion injection that can fulfill the prerequisites for injection preparations with a simple method and simple reconstitution before use. All combinations of the oil and aqueous phases were formed using the self-emulsification technique. The optimum stability was estimated on particle size, polydispersity index (PDI), and zeta potential at 5°C, 25°C, and 40°C for 120 days. Based on the minimum amount of Tween 80, drug loading, and the highest transmittance value, the optimal microemulsion formula was obtained at a concentration of 2.8% (1:1 combination of olive oil and medium-chain triglyceride oil), 11.2% Tween 80, 2.8% propylene glycol, and 83.2% water for injection. The optimal formula has a droplet size of 16.7 ± 11.2 nm, a zeta potential of 0.8 mV, and a PDI of 0.196. The formula was stable at 45°C storage for up to 14 days, at 25°C, and at 5°C, steady at 120 days based on droplet size diameter of 21.27 ± 0.00 nm, zeta potential -1.3 ± 0.10 mV, and PDI 0.38 ± 0.02. The optimum formula by simple reconstitution without additional isotonization has fulfilled the requirements for injection preparation. It was concluded that the ibuprofen microemulsion with a combination of medium-chain triglyceride oil and olive oil could form a stable microemulsion and has the potential to be developed as a pharmaceutical preparation in the form of injectable ibuprofen.

Medium chain triglycerides (MCT): State-of-the-art on chemistry, synthesis, health benefits and applications in food industry.

Medium chain triglycerides (MCT) are esters of fatty acids with 6 to 12 carbon atom chains. Naturally, they occur in various sources; their composition and bioactivity are source and extraction process-linked. The molecular size of MCT oil permits unique metabolic pathways and energy production rates, making MCT oil a high-value functional food. This review details the common sources of MCT oil, presenting critical information on the various approaches for MCT oil extraction or synthesis. Apart from conventional techniques, non-thermal processing methods that show promising prospects are analyzed. The biological effects of MCT oil are summarized, and the range of need-driven modification approaches are elaborated. A section is devoted to highlighting the recent trends in the application of MCT oil for food, nutraceuticals, and allied applications. While much is debated about the role of MCT oil in human health and wellness, there is limited information on daily requirements, impact on specific population groups, and effects of long-term consumption. Nonetheless, several studies have been conducted and continue to identify the most effective methods for MCT oil extraction, processing, handling, and storage. A knowledge gap exists and future research must focus on technology packages for scalability and sustainability.

Triheptanoin (Dojolvi)

&#x0D; CADTH recommends that Dojolvi should be reimbursed by public drug plans as a source of calories and fatty acids for the treatment of adult and pediatric patients with a long-chain fatty acid oxidation disorders (LC-FAOD) if certain conditions are met.&#x0D; Dojolvi should only be covered to treat patients with and without a confirmed LC-FAOD diagnosis if they present with acute life-threatening symptoms that are consistent with those of LC-FAOD. Dojolvi should only be covered in patients who need a different treatment than over-the-counter even-chain medium-chain triglyceride (MCT) oil.&#x0D; Dojolvi should only be reimbursed if prescribed by clinicians experienced in the management of LC-FAOD and if the price of Dojolvi is reduced. Reimbursement of Dojolvi should be renewed every 12 months if individual patient treatment goals are being met.&#x0D;

Chemical Emissions From Heated Vitamin E Acetate-Insights to Respiratory Risks From Electronic Cigarette Liquid Oil Diluents Used in the Aerosolization of Δ9-THC-Containing Products.

As of February 18, 2020, the e-cigarette, or vaping, product use associated lung injury (EVALI) outbreak caused the hospitalization of a total of 2,807 patients and claimed 68 lives in the United States. Though investigations have reported a strong association with vitamin E acetate (VEA), evidence from reported EVALI cases is not sufficient to rule out the contribution of other chemicals of concern, including chemicals in either THC or non-THC products. This study characterized chemicals evolved when diluent oils were heated to temperatures that mimic e-cigarette, or vaping, products (EVPs) to investigate production of potentially toxic chemicals that might have caused lung injury. VEA, vitamin E, coconut, and medium chain triglyceride (MCT) oil were each diluted with ethanol and then tested for constituents and impurities using a gas chromatograph mass spectrometer (GC/MS). Undiluted oils were heated at 25°C (control), 150°C, and 250°C in an inert chamber to mimic a range of temperatures indicative of aerosolization from EVPs. Volatilized chemicals were collected using thermal desorption tubes, analyzed using a GC/MS, and identified. Presence of identified chemicals was confirmed using retention time and ion spectra matching with analytic standards. Direct analysis of oils, as received, revealed that VEA and vitamin E were the main constituents of their oils, and coconut and MCT oils were nearly identical having two main constituents: glycerol tricaprylate and 2-(decanoyloxy) propane-1,3-diyl dioctanoate. More chemicals were measured and with greater intensities when diluent oils were heated at 250°C compared to 150°C and 25°C. Vitamin E and coconut/MCT oils produced different chemical emissions. The presence of some identified chemicals is of potential health consequence because many are known respiratory irritants and acute respiratory toxins. Exposure to a mixture of hazardous chemicals may be relevant to the development or exacerbation of EVALI, especially when in concert with physical damage caused by lung deposition of aerosols produced by aerosolizing diluent oils.

Size effects of discoidal PLGA nanoconstructs in Pickering emulsion stabilization

AbstractSolid particle stabilized emulsions, using unique shape defined particles, are receiving increasing research interest due to ease of formulation and interesting physiochemical characteristics. There is, however, a need to systematically investigate the effect of anisotropic discoidal microparticles, realized with top‐down fabrication approaches, in emulsion stabilization. Here, the effect of poly(d,l‐lactide‐co‐glycolide) (PLGA) discoidal polymeric nanoconstruct (DPN) size on the formation and stability of oil‐in‐water emulsions is studied. Particles with a diameter of 1, 2, and 5 μm are fabricated with a lithographic templating technique, and used to stabilize medium chain triglyceride (MCT) oil emulsions. Three phase contact angles decreased from 85° ± 7° to 68° ± 12° moving from 1 to 5 μm DPN stabilized emulsions, showing a particle “hydrophilicity” increase with size. Microscopy imaging showed that the mean droplet diameter and dispersity increased with particle size, and that DPNs were present at the oil–water interface. DPN based emulsions were stable for about 24 h or less in the case of 1 and 2 μm DPNs. Emulsion stability was shorter than 12 h in case of 5 μm DPNs. Finally, calculations of DPN detachment free energies ΔGdw and excess surface coverages Cexcess demonstrated that, despite the significantly high adhesion energy of the discoidal DPN, emulsion stability was mostly affected by gravitational forces for DPN sizes above 2 μm. The use of PLGA and MCT oil in this study is relevant for future use of Pickering emulsions in pharmaceutical and drug delivery applications.

Pickering emulsions stabilized by biodegradable dextran-based nanoparticles featuring enzyme responsiveness and co-encapsulation of actives

In this study, Pickering emulsions of dodecane and medium chain triglyceride (MCT) oils were stabilized by simply alkylated-dextran nanoparticles. Our findings show that very little of these bio-friendly nanoparticles is necessary to stabilize Pickering emulsions while providing a high time stability (more than a year at 37 °C). As dextran is known to be cleavable by dextranase enzyme, hydrolysis of the nanoparticles in the presence of dextranase could be achieved. This allowed performing on-demand destabilization of Pickering emulsions. Furthermore, two different fluorescent probes were loaded into the stabilizing particles and the oil droplets respectively, providing a proof of concept for co-encapsulation of actives in advanced delivery applications. Additionally, to a conventional fluorescence probe, quinine, an antimalarial drug was also encapsulated into the nanoparticles.

Cyclodextrin-based Pickering nanoemulsions containing amphotericin B: Part I. evaluation of oil/cyclodextrin and amphotericin B/cyclodextrin inclusion complexes

Prior to developing cyclodextrin (CD) stabilized Pickering nanoemulsions (NEs) containing amphotericin B (AmB) for fungal keratitis therapy, the AmB solubility and the effect of CDs on complex formation of medium chain triglyceride (MCT) oils and AmB was investigated. The phase-solubility profiles of AmB in various aqueous CD solutions were determined. The AmB solubilization in oils and surfactants was also examined. Two CDs, γ-cyclodextrin (γCD) and 2-hydroxypropyl-γCD (HPγCD), along with MCT oil and lecithin were chosen because of their high solubilization ability for AmB. The solution- and solid-state characterizations of their inclusion complexes were investigated. Proton nuclear magnetic resonance studies demonstrated the formation of MCT oil/α-cyclodextrin (αCD), AmB/γCD, and AmB/HPγCD complexes. Some interactions between AmB and CDs were observed by solid-state characterizations using Fourier-transform infrared (FT-IR) spectroscopy, powder X-ray diffractometry and differential scanning calorimetry. The interfacial tension at the oil-water interface, the analysis of dissolved CD content in saturated MCT oil, and FT-IR confirmed the affinity of αCD for MCT oil. Scanning electron microscopy revealed that the MCT oil/αCD complexes can transformed into the precipitated solid complexes at high αCD concentration. Thus, the MCT oil/αCD complexes will be used as solid particles to stabilize Pickering NEs for further studies.

Use of medium chain triglyceride (MCT) oil in subjects with Alzheimer's disease: A randomized, double-blind, placebo-controlled, crossover study, with an open-label extension.

IntroductionCerebral glucose and insulin metabolism is impaired in Alzheimer's disease (AD). Ketones provide alternative energy. Will medium chain triglyceride (MCT) oil, a nutritional source of ketones, impact cognition in AD?MethodsThis was a 6‐month randomized, double‐blind, placebo‐controlled, crossover study, with 6‐month open‐label extension in probable AD subjects, on stable medications. MCT dose was 42 g/day, or maximum tolerated. Cognition was assessed with Mini‐Mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), and Cognigram®.ResultsTwenty subjects, average age 72.6 years, 45% women, 70% university educated had baseline MMSE 22.6/30 (10–29); MoCA 15.6/30 (4–27); baseline Cognigram® Part 1: 65–106, Part 2: 48–107. Average MCT oil consumption was 1.8 tablespoons/day (25.2 g, 234 kcal). Eighty percent remained stable or improved. Longer MCT exposure and age > 73, resulted in higher final MMSE (P < .001) and Cognigram® 1 scores.DiscussionThis is the longest duration MCT AD study to date. Eighty percent had stabilization or improvement in cognition, and better response with 9‐month continual MCT oil.

STORAGE STUDIES OF FUNCTIONAL YOGURT DRINK MADE WITH MEDIUM CHAIN TRIGLYCERIDE OIL AND ITS QUALITY ATTRIBUTES

Medium chain triglyceride oil based functional yogurt drink was prepared from skim milk with the addition of MCT at two different variations (0.5 and 1.0%) and transferred to a sterilized glass bottle with skimmed milk at (43 °C). The pH and TSS values of yogurt decreased over the storage period with a simultaneous increase in bacterial count. The sample with concentration of 1 % of MCT treated at 43 °C showed the best results compared to control samples with an acceptable microbial load. The highest lightness index was found in MY2 with 82.30 and lowest was in MY1 with 80.5 at the 1st day of the storage. At the end of the storage period (21st day), decreased lightness (L*) was observed in MY1 followed by control sample. TA was significantly increased in MYC than in MCT samples with value of 1.11±0.11% and 1.10±0.10% respectively. Acceptability index of sensory attributes of developed yogurt drink of 1 % MCT oil incorporated shows the best overall acceptability compared to other samples. Hence it is a potential replacement to the probiotic drinks with added health benefits.

Hemostatic rebalance in neonatal intrahepatic cholestasis with citrin deficiency.

Neonatal intrahepatic cholestasis with citrin deficiency (NICCD) results in coagulopathy due to decreased levels of vitamin (V)K-dependent clotting factors, similar to biliary atresia (BA). However, the involvement of VK-independent coagulant and anticoagulant factor(s) remains unknown. We examined relationships between coagulant and anticoagulant potential before and after nutritional treatment in NICCD. Three cases (aged 12, 21, and 45 days) with NICCD-associated coagulopathy were evaluated with standard coagulation/anticoagulation tests and comprehensive coagulation assays, rotational thromboelastometry, and protein C/protein S (PC/PS) pathway function assay (ThromboPath® ), before and after nutritional treatment. In all cases, activated partial thromboplastin time and prothrombin time were significantly prolonged, which is associated with very low levels of VK-independent fibrinogen and antithrombin. The initiation of nutritional treatment of medium-chain triglycerides oil improved these levels within the normal range, although low levels of other clotting factors were modestly increased. Whole blood- rotational thromboelastometry analysis revealed near-normal coagulation potential, even before treatment, comparable to healthy adults, and supportive of their non-bleeding symptoms. The introduction of nutritional treatment had further improved comprehensive coagulation potential. The global PC/PS-pathway function assay demonstrated the absence of the features of this function associated with the pathogenesis of NICCD. Compared to BA, the plasma levels of fibrinogen and antithrombin in all cases were markedly low, whilst those after treatment improved, especially to similar level of BA. Neonatal intrahepatic cholestasis with citrin deficiency has the characteristic of rebalancing hemostatic mechanisms associated with coagulant and anticoagulant potential involving low levels of fibrinogen and antithrombin, suggesting a pathophysiological coagulopathy distinct from BA.

Tobacco Hornworm as an Insect Model System for Cannabinoid Pre-clinical Studies.

With increased attention on cannabinoids in medicine, several mammalian model organisms have been used to elucidate their unknown pharmaceutical functions. However, many difficulties remain in mammalian research, which necessitates the development of non-mammalian model organisms for cannabinoid research. The authors suggest the tobacco hornworm Manduca sexta as a novel insect model system. This protocol provides information on preparing the artificial diet with varying amounts of cannabidiol (CBD), setting up a cultivation environment, and monitoring their physiological and behavioral changes in response to CBD treatment. Briefly, upon receiving hornworm eggs, the eggs were allowed 1-3 days at 25 °C on a 12:12 light-dark cycle to hatch before being randomly distributed into control (wheat germ-based artificial diet; AD), vehicle (AD + 0.1% medium-chain triglyceride oil; MCT oil) and treatment groups (AD + 0.1% MCT + 1 mM or 2 mM of CBD). Once the media was prepared, 1st instar larvae were individually placed in a 50 mL test tube with a wooden skewer stick, and then the test tube was covered with a cheesecloth. Measurements were taken in 2-day intervals for physiological and behavioral responses to the CBD administration. This simple cultivation procedure allows researchers to test large specimens in a given experiment. Additionally, the relatively short life cycles enable researchers to study the impact of cannabinoid treatments over multiple generations of a homogenous population, allowing for data to support an experimental design in higher mammalian model organisms.