Metabolomic and biochemical changes in the plasma and liver of toxic milk mice model of Wilson disease.

Wilson disease (WD) is an inherited disorder characterized by abnormal copper metabolism with complex pathological features. Currently, this mechanism of copper overload-induced hepatic injury remains unclear. In this study, male toxic milk (TX) mice were selected as experimental subjects. Copper levels and biochemical indices were measured by atomic absorption spectroscopy (AAS) and kits. Liver tissue ultrastructure was observed by hematoxylin-eosin (H&E), sirius red staining and transmission electron microscopy. Plasma and liver metabolic profiles of TX mice were characterized by untargeted metabolomics. In addition, the expression of enzymes related to arachidonic acid metabolism in liver tissue was detected by Western blotting. The results showed the excessive copper content, concomitant oxidative stress, and hepatic tissue structural damage in TX mice. Seventy-eight metabolites were significantly different in WD, mainly involved in the metabolism of arachidonic acid, glycerophospholipids, sphingolipids, niacin and nicotinamide, and phenylalanine. Furthermore, the arachidonic acid metabolic pathway is an important pathway involved in WD metabolism. The level of arachidonic acid in the liver of TX mice was significantly lower (p < 0.01) compared to the control group. The expression of cytoplasmic phospholipase A2 (cPLA2) and arachidonic acid 12-lipoxygenase (ALOX12), related to the arachidonic acid metabolic pathway, was significantly different in the liver of TX mice (p < 0.01). Modulation of the arachidonic acid metabolic pathway could be a potential therapeutic strategy to alleviate WD symptoms.

Silicon Ether Ionizable Lipids Enable Potent mRNA Lipid Nanoparticles with Rapid Tissue Clearance.

The advent of mRNA for nucleic acid (NA) therapeutics has unlocked many diverse areas of research and clinical investigation. However, the shorter intracellular half-life of mRNA compared with other NAs may necessitate more frequent dosing regimens. Because lipid nanoparticles (LNPs) are the principal delivery system used for mRNA, this could lead to tolerability challenges associated with an accumulated lipid burden. This can be addressed by introducing enzymatically cleaved carboxylic esters into the hydrophobic domains of lipid components, notably, the ionizable lipid. However, enzymatic activity can vary significantly with age, disease state, and species, potentially limiting the application in humans. Here we report an alternative approach to ionizable lipid degradability that relies on nonenzymatic hydrolysis, leading to a controlled and highly efficient lipid clearance profile. We identify highly potent examples and demonstrate their exceptional tolerability in multiple preclinical species, including multidosing in nonhuman primates (NHP).

The dosing regimen for 17-hydroxyprogesterone caproate was suboptimal: lessons for future pharmacotherapy for pregnant women.

BACKGROUND: Makena (17-hydroxyprogesterone caproate) was approved by the United States Food and Drug Administration for the prevention of recurrent spontaneous preterm birth in 2011 under the accelerated approval pathway, but fundamental pharmacokinetic or pharmacodynamic (Phase 1 and Phase 2) studies were not performed. At the time, there were no dose-response or concentration-response data. The therapeutic concentration was not known. The lack of such data brings into question the dosing regimen for 17-hydroxyprogesterone caproate and if it was optimized. OBJECTIVE: The purpose of this study was to evaluate the dosing regimen for 17-hydroxyprogesterone by analyzing 3 data sets in which the 17-hydroxyprogesterone caproate pharmacology was evaluated, namely the Maternal-Fetal Medicine Omega 3 study, the Obstetric-Fetal Pharmacology Research Units study, and the Obstetrical-Fetal Pharmacology Research Centers study. If an inappropriate dosing regimen could be identified, such information could inform future studies of pharmacotherapy in pregnancy. STUDY DESIGN: Data from the Omega 3 study were used to determine if plasma concentration was related to spontaneous preterm birth risk and if a threshold concentration could be identified. Data from the Obstetric-Fetal Pharmacology Research Units study were used to determine the half-life of 17-hydroxyprogesterone caproate and to develop a model to simulate drug concentrations with various dosing regimens. Data from the Obstetrical-Fetal Pharmacology Research Centers study were used to determine the relationship between dose and safety outcomes. RESULTS: Analysis of the Omega 3 data set indicated that the risk for spontaneous preterm birth decreased as the log concentration of 17-hydroxyprogesterone caproate increased (odds ratio, 0.04; 95% confidence interval, 0.00-0.90). A steady state concentration of >9 ng/mL (equivalent to >8 ng/mL at 25-28 weeks) was associated with the lowest risk for spontaneous preterm birth (hazard ratio, 0.52; 95% confidence interval, 0.27-0.98; P=.04); this concentration was not achieved in 25% of subjects who received the 250 mg weekly dose. In the Obstetrical-Fetal Pharmacology Research Units study, the adjusted half-life (median and interquartile range) of 17-hydroxyprogesterone caproate was 14.0 (11.5-17.2) days. Simulations indicated that with the 250 mg weekly dose, >5 weekly injections were required to reach the 9 ng/mL target; however, those with the shortest half-life (corresponding to higher clearance), never reached the targeted 9 ng/mL concentration. In 75% of subjects, a loading dose of 500 mg weekly for 2 weeks followed by 250 mg weekly achieved and maintained the 9 ng/mL concentration within 2 weeks but in those 25% with the shortest half-life, concentrations exceeded the 9 ng/mL target for only 3 weeks. In the Obstetrical-Fetal Pharmacology Research Centers study, all 65 subjects who received a weekly dose of 500 mg exceeded the 9 ng/mL steady state. CONCLUSION: The dosing regimen for 17-hydroxyprogesterone caproate was inadequate. There is a significant inverse relationship between drug concentration and spontaneous preterm birth. The risk was lowest when the concentration exceeded 9 ng/mL, but 25% of women who received the 250 mg weekly dose never reached or maintained this concentration. The drug's long half-life necessitates a loading dose to achieve therapeutic concentrations rapidly. The omission of basic pharmacologic studies to determine the proper dosing may have compromised the effectiveness of 17-hydroxyprogesterone caproate. Future pharmacotherapy trials in pregnancy must first complete fundamental pharmacology studies.

Simplified processing and rapid quantification of buprenorphine, norbuprenorphine, and their conjugated metabolites in human plasma using UPLC-MS/MS: Assessment of buprenorphine exposure during opioid use disorder treatment.

Opioid use disorder (OUD) is a chronic neurobehavioral ailment and is prevalent in pregnancy. OUD is commonly treated with methadone or buprenorphine (BUP). Pregnancy is known to alter the pharmacokinetics of drugs and may lead to changes in drug exposure and response. A simple, specific, and sensitive analytical method for measuring the parent drug and its metabolites is valuable for assessing the impact of pregnancy on drug exposure. A new liquid chromatography-tandem mass spectrometric method that utilized a simple protein precipitation procedure for sample preparation and four deuterated internal standards for quantification was developed and validated for BUP and its major metabolites (norbuprenorphine [NBUP], buprenorphine-glucuronide [BUP-G], and norbuprenorphine-glucuronide [NBUP-G]) in human plasma. The standard curve was linear over the concentration range of 0.05-100 ng/mL for BUP and NBUP, and 0.1-200 ng/mL for BUP-G and NBUP-G. Intra- and inter-day bias and precision were within ±15% of nominal values for all the analytes. Quality controls assessed at four levels showed high recovery consistently for all the analytes with minimal matrix effect. Adequate analyte stability was observed at various laboratory conditions tested. Overall, the developed method is simple, sensitive, accurate and reproducible, and was successfully applied for the quantification of BUP and its metabolites in plasma samples collected from pregnant women in a clinical study assessing BUP exposure during OUD treatment.

Simultaneous quantitation of ketamine, norketamine and dehydronorketamine in human milk using a novel ultra high-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) assay.

There is a paucity of data on the transfer of ketamine from maternal blood into human milk. Quantification of ketamine in human milk provides information about the potential exposure of the infant to ketamine and its metabolites from the mother during lactation. A highly specific, reproducible, and sensitive UPLC-MS/MS based analytical method was developed and validated for the quantitation of ketamine and its metabolites (norketamine and dehydronorketamine) in human milk. Samples were subjected to a simple protein precipitation and ketamine-d4 and norketamine-d4 were used as internal standards. Separation of the analytes was achieved using an Acquity UPLC equipped with BEH RP18 1.7 µm, 2.1 × 100 mm column. Mass spectrometric analysis of the analyte ions was carried out using electrospray with positive ionization and multiple reaction monitoring mode. The assay was linear over a concentration range of 1-100 ng/mL for ketamine and norketamine, and 0.1-10 ng/mL for dehydronorketamine. Acceptable intra-day and inter-day accuracy and precision were observed for all the analytes. High recovery of the analytes and minimal matrix effect were observed. Stability of analytes was confirmed at the tested conditions. This assay was successfully used to measure analytes in human milk samples collected from lactating women enrolled in a clinical research study. This is the first validated method that simultaneously quantified ketamine and its metabolites in human milk.

The rate of orthokeratology lens use and associated factors in 33,280 children and adolescents with myopia: a cross-sectional study from Shanghai.

OBJECTIVES: To investigate the rate of orthokeratology lens (ortho-k lens) use and its associated factors in children and adolescents with myopia. METHODS: Cross-sectional study. Children from 104 primary and middle schools in Shanghai were enrolled by cluster sampling. Ophthalmic examinations were conducted and information was obtained using questionnaires for associated factors analysis. RESULTS: A total of 72,920 children and adolescents were included, among which 32,259 were the potential population for ortho-k lens use. A total of 1021 participants used ortho-k lenses, equating to a use rate of 1.4% in the total population and 3.1% in the potential population. Age (OR 0.91, 95% CI: 0.88-0.95, p < 0.001), BMI (≥95th percentile: OR 0.48, 95% CI: 0.35-0.66, p < 0.001), age at initiation of refractive correction (≤12 years: OR 1.75, 95% CI: 1.31-2.33, p < 0.001), and parental myopia (either: OR 2.09, 95% CI: 1.58-2.75, p < 0.001; both: OR 3.94, 95% CI: 3.04-5.11, p < 0.001) were independently associated with ortho-k lens use. Of the ortho-k lenses users, 12.4% had a logMAR CVA of ≥0.3. A correction target (SE) of ≤-3.0 D (OR 2.05, 95% CI: 1.38-3.05, p < 0.001) and a sleeping duration of ≤6 h (OR 4.19, 95% CI: 2.03-8.64, p < 0.001) were factors independently associated with CVA ≥ 0.3. CONCLUSIONS: A certain proportion of children and adolescents in Shanghai chose to wear ortho-k lenses, related to the situation of parents and children themselves. Health education and follow-ups should be strengthened to ensure orthokeratology application quality.

Effect of formulation and route of administration on the distribution of 17-hydroxyprogesterone caproate in rats.

Weekly intramuscular (250 mg/week) or subcutaneous (275 mg/week) injections of 17-hydroxyprogesterone caproate (17-OHPC) is the only treatment option for the prevention of preterm birth in women with a prior history of preterm delivery.The objective of the current study was to determine the relative distribution of 17-OHPC in selected tissues in adult female SD rats after IM (oily formulation or solution), IV (solution), PO (solution), or intravaginal (suppository) administration.Plasma, uterus, adipose, and liver samples were collected at various times and analysed by LC-MS-MS.The highest concentrations of 17-OHPC were observed in the adipose tissue, after IM (oily formulation), and intravaginal administration.Substantial concentrations of 17-OHPC were also observed in the uterus after IM, intravaginal and IV administration.17-OHPC was not detected in the liver and in any of the tissues tested after PO administration.17-OHPC levels in plasma after intravaginal suppository administration were low despite substantial concentrations in the adipose and the uterus.The distribution of 17-OHPC depends on the formulation, the route of administration, and the sampling time.Low systemic concentrations and substantial distribution in the tissues of interest after intravaginal administration warrants future studies to evaluate the potential of the daily intravaginal route of administration of 17-OHPC.

Changes of corneal biomechanics in children using orthokeratology and their roles in predicting axial length progression-A prospective 2-year study.

PURPOSE: To determine how orthokeratology (ortho-k) affects corneal biomechanical properties in myopia control and whether corneal biomechanical parameters can predict clinical efficacy of ortho-k. METHODS: A total of 125 children 7-15 years of age using ortho-k lenses were followed in this clinical practice and data of their right eyes were analysed. Corneal biomechanical parameters and most ocular biometry were measured at baseline, 1 week, and at 1, 3, 6, 12, 18 and 24 months. Axial length (AL) was collected every 6 months after baseline measurements. RESULTS: During the 2-year follow up, nine corneal biomechanical parameters, including deformation amplitude maximum (DA), varied between baseline and 1 week (p < 0.05) and stabilized during the rest of wearing period (p > 0.05). The mean AL increased from 25.02 ± 0.84 mm to 25.38 ± 0.81 mm and baseline DA strongly correlated with AL progression (Pearson r = 0.37). In the multiple regression models, baseline age, AL and DA were the independent factors for AL progression (R2 : 0.7849, 0.2180 in low and moderate myopes). The area under the receiver operating characteristic curves using the three variables for predicting excessive AL progression (>0.35 mm during 2 years) in low and moderate myopes was 0.902 and 0.698. CONCLUSIONS: Corneal biomechanics firstly fluctuated before becoming stable with long-term ortho-k use. Corneal biomechanics was associated with AL progression in children wearing ortho-k lenses. DA combined with age and AL at baseline could predict AL progression in low myopes using ortho-k.

Protective effect of curcumin on hepatolenticular degeneration through copper excretion and inhibition of ferroptosis.

BACKGROUND: Hepatolenticular degeneration (HLD) is an autosomal recessive disorder concerning copper metabolism. Copper overload is also accompanied by iron overload in HLD patients, which can lead to ferroptosis. Curcumin, the active component in turmeric, has the potential to inhibit ferroptosis. PURPOSE: The current study proposed a systematic investigation of the protective effects of curcumin against HLD and the underlying mechanisms. METHODS: The protective effect of curcumin on toxic milk (TX) mice was studied. Liver tissue was observed via hematoxylin-eosin (H&E) staining and the ultrastructure of the liver tissue was observed through transmission electron microscopy. Copper levels in the tissues, serum, and metabolites were measured by atomic absorption spectrometry (AAS). In addition, serum and liver indicators were evaluated. In cellular experiments, the effect of curcumin on the viability of rat normal liver cells (BRL-3A) was determined via the 3-[4,5-dimethylthiazol-2-yl)]-2,5-diphenyltetrazolium bromide (MTT) assay. Cell and mitochondrial morphology were observed in curcumin-mediated HLD model cells. The intracellular copper ion fluorescence intensity was observed via fluorescence microscopy, and intracellular copper iron content was detected using AAS. Further, oxidative stress indicators were evaluated. Cellular reactive oxygen species (ROS) and cellular mitochondrial membrane potential were examined via flow cytometry. Furthermore, the expression levels of nuclear factor erythroid-2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and glutathione peroxidase 4 (GPX4) were determined via western blotting (WB). RESULTS: The histopathology of the liver confirmed the hepatoprotective effects of curcumin. Curcumin improved copper metabolism in TX mice. Both serum liver enzyme markers and antioxidant enzyme levels indicated the protective effect of curcumin against HLD-related liver injury. The MTT assay results showed that curcumin was protective against excess copper-induced injury. Curcumin improved the morphology of HLD model cells and their mitochondrial morphology. The Cu2+ fluorescent probe and the AAS results indicated that curcumin reduced Cu2+ content in HLD hepatocytes. In addition, curcumin improved oxidative stress levels and prevented the decline of mitochondrial membrane potential in HLD model cells. The ferroptosis inducer Erastin reversed these effects of curcumin. WB revealed that curcumin promoted Nrf2, HO-1, and GPX4 protein expression in HLD model cells, and the Nrf2 inhibitor ML385 reversed the effects of curcumin. CONCLUSION: Curcumin demonstrates a protective role by expelling copper and inhibiting ferroptosis, activating the Nrf2/HO-1/GPX4 signaling pathway in HLD.

Assessment of glomerular filtration and tubular secretion in baboons with life-supporting pig kidney grafts.

With pig kidney xenotransplantation nearing clinical reality, it is imperative to measure pig kidney function in the graft recipients. Our aims were (i) to compare inulin clearance after a short intravenous (IV) bolus with steady-state inulin IV infusion, (ii) to use this method to measure the glomerular filtration rate (GFR), and (iii) to determine the tubular secretory function using cefoxitin in a pig-to-baboon renal transplant model. A short IV infusion of inulin and cefoxitin were followed by a maintenance IV infusion of inulin over 5 h in seven healthy baboons, three healthy pigs, and five baboons after bilateral native nephrectomy and intra-abdominal pig renal transplantation. Blood and urine samples were collected. Serum and urinary inulin and serum cefoxitin concentrations measured by validated assays were used to calculate GFR and renal secretion. GFR calculated were similar by both methods. The body weight normalized total body clearance of inulin was similar in pigs and baboons despite differences in absolute clearances. Pig kidney transplanted into baboons provided similar clearance in baboons when normalized to baboon body weight and sustained filtration and secretory functions. The study documented that pig kidneys support the physiologic needs of baboons and are likely to support human recipients as well.

Unsaturated, Trialkyl Ionizable Lipids are Versatile Lipid-Nanoparticle Components for Therapeutic and Vaccine Applications.

Lipid nanoparticles (LNPs) have proven a successful platform for the delivery of nucleic acid (NA)-based therapeutics and vaccines, with the ionizable lipid component playing a key role in modulating potency and tolerability. Here, a library of 16 novel ionizable lipids is screened hypothesizing that short, branched trialkyl hydrophobic domains can improve LNP fusogenicity or endosomal escape, and potency. LNPs formulated with the top-performing trialkyl lipid (Lipid 10) encapsulating transthyretin siRNA elicit significantly greater gene silencing and are better tolerated than those with the benchmark Onpattro lipid DLin-MC3-DMA. Lipid 10 also demonstrates superior liver delivery of mRNA when compared to other literature ionizable lipids, is well tolerated, and successfully repeat-doses in nonhuman primates. In a prime-boost hemagglutinin rodent vaccine model, intramuscular administration of Lipid-10 LNP elicits comparable or better antibody titers to the SM-102 and ALC-0315 lipid compositions used in the U.S. Food and Drug Administration approved mRNA COVID vaccines. These data suggest that Lipid 10 is a particularly versatile ionizable lipid, well-suited for both systemic therapeutic and intramuscular vaccine applications and able to successfully deliver diverse NA payloads.

Accommodation and vergence function in children using atropine combined with orthokeratology.

PURPOSE: This study aimed to evaluate binocular vision in terms of vergence and accommodative measurements in children treated with 0.01% atropine combined with orthokeratology (OK). METHODS: This was a prospective and randomized controlled clinical trial involving participants aged 8 to 12 years, with a spherical equivalent (SE) ranging from - 1.00 to - 6.00D. Participants were randomly divided into four groups: 1) a combination group using 0.01% atropine solution and OK lens; 2) an OK group using placebo solution and OK lens; 3) an atropine group using 0.01% atropine solution and wearing spectacles; and 4) a control group using placebo solution and wearing spectacles. Binocular vision was determined at baseline and at 3-month visits, with evaluations including horizontal phoria, fusional vergence, the accommodative convergence/accommodation (AC/A) ratio, accommodative lag, and accommodative amplitude (AA). The Wilcoxon signed-rank test was used to compare the changes in binocular vision in each group, and the Kruskal-Wallis test was used for comparisons of four groups. RESULTS: Sixty-two participants completed the study. There was no significant difference in baseline refraction, accommodation or vergence measurements among the groups (all P > 0.05). Three months later, the accommodative lag significantly decreased in the OK group (P = 0.002) but remained unchanged in the other three groups (all P > 0.05). In addition, binocular accommodative facilities and positive relative accommodations increased in the combination and OK groups (both P < 0.05) but remained unchanged in the atropine and control groups (both P > 0.05). Only the participants with esophoria in the OK group had a significant decrease in esophoria (P = 0.008). Moreover, the changes in fusional vergence and AC/A did not significantly differ between the four groups (all P > 0.05). CONCLUSION: Accommodative measurements changed similarly in the groups treated with OK. Changes in vergence measurements after treatment with 0.01% atropine were not significant.

Effect of atropine, orthokeratology and combined treatments for myopia control: a 2-year stratified randomised clinical trial.

PURPOSE: To investigate the 2-year efficacy of atropine, orthokeratology (ortho-k) and combined treatment on myopia. To explore the factors influencing the efficacy. METHODS: An age-stratified randomised controlled trial. Children (n=164) aged 8-12 years with spherical equivalent refraction of -1.00 to -6.00 D were stratified into two age subgroups and randomly assigned to receive placebo drops+spectacles (control), 0.01% atropine+spectacles (atropine), ortho-k+placebo (ortho-k) or combined treatment. Axial length was measured at baseline and visits at 6, 12, 18 and 24 months. The primary analysis was done following the criteria of intention to treat, which included all randomised subjects. RESULTS: All interventions can significantly reduce axial elongation at all visits (all p<0.05). Overall, the 2-year axial elongation was significantly reduced in combined treatment than in monotherapies (all p<0.05). After stratification by age, in the subgroup aged 8-10, the difference between combined treatment and ortho-k became insignificant (p=0.106), while in the subgroup aged 10-12, the difference between combined treatment and atropine became insignificant (p=0.121). A significant age-dependent effect existed in the ortho-k group versus the control group (p for interaction=0.013), and a significant age-dependent effect existed in the ortho-k group versus the atropine group (p for interaction=0.035), which indicated that ortho-k can achieve better efficacy in younger children. CONCLUSIONS: Atropine combined with ortho-k treatment can improve the efficacy of myopia control compared with monotherapy in children aged 8-12. Younger children might benefit more from ortho-k. TRIAL REGISTRATION NUMBER: ChiCTR1800015541.

Variability of Accommodative Microfluctuations in Myopic and Emmetropic Juveniles during Sustained near Work.

Near work has been considered to be a potential risk factor for the onset of myopia, but with inadequate evidence. Chinese adolescents use digital devices for near work, such as study and entertainment purposes, especially during the COVID-19 pandemic. In this study, we investigated the influence of prolonged periods of near work on accommodative response, accommodative microfluctuations (AMFs), and pupil diameter between juvenile subjects of myopia and emmetropia. Sixty juveniles (30 myopes and 30 emmetropes) were recruited for the study. Participants were instructed to play a video game on a tablet PC at a distance of 33.3 cm for 40 min. Accommodative response and pupil diameter were measured with an open-field infrared refractometer in High-speed mode. Parameters of the subjects were measured once every 10 min, and analyzed by one-way repeated measure ANOVA for variation tendency. There were no significant differences between emmetropia and myopia groups with respect to age and sex (p > 0.05). The low-frequency component (LFC) of myopia gradually increased with time, reached a peak at 30 min, and then declined (p = 0.043). The high-frequency component (HFC) of myopia also reached a peak at 30 min (p = 0.036). Nevertheless, there was no significant difference in the LFC (p = 0.171) or HFC (p = 0.278) of the emmetropia group at each time point. There was no significant difference in the mean and standard deviation of the accommodative response and pupil diameter both in emmetropic and myopic juveniles. Compared with juvenile emmetropes, myopes exhibit an unstable tendency in their accommodation system for prolonged near work at a certain time point. Accommodative microfluctuations may be a sensitive, objective indicator of fatigue under sustained near work in juvenile myopes.

Protective Mechanism of Gandou Decoction in a Copper-Laden Hepatolenticular Degeneration Model: In Vitro Pharmacology and Cell Metabolomics.

Gandou decoction (GDD) is a classic prescription for the treatment of hepatolenticular degeneration (HLD) in China; however, the liver-protecting mechanism of this prescription needs further evaluation. In the present study, we explored the protective mechanisms of GDD in a copper-laden HLD model using integrated pharmacology and cellular metabolomics in vitro. The results revealed that GDD could significantly promote copper excretion in copper-laden HLD model cells and improve the ultrastructural changes in hepatocytes. In addition, GDD could decrease the extent of lipid peroxidation, levels of reactive oxygen species, and the release rate of lactate dehydrogenase while increasing the activity of superoxide dismutase and the ratio of glutathione to oxidized glutathione in the copper-laden HLD model cells. On conducting statistical analysis of significant metabolic changes, 47 biomarkers and 30 related metabolic pathways were screened as pharmacological reactions induced by GDD in HLD model cells. d-glutamate and d-glutamine metabolic pathways showed the highest importance and significance among the 30 metabolic pathways, and the differential expression levels of the glutamine synthetase (GS) and the renal type and liver type GLS (GLS1 and GLS2) proteins were verified by Western blotting. Collectively, our data established the underlying mechanism of GDD therapy, such as the promotion of copper excretion and improvement in oxidative stress by regulating the expressions of GS, GLS1, and GLS2 protein to protect hepatocytes from injury.

Liver and urine metabolomics reveal the protective effect of Gandou decoction in copper-laden Hepatolenticular degeneration model rats.

Hepatolenticular degeneration (HLD) is an inherited disorder associated with human copper metabolism. Gandou decoction (GDD), a traditional Chinese medicinal formula, has been used as a therapeutic agent for the treatment of HLD in China for decades. Recent pharmacological evaluation in our laboratory has demonstrated that GDD exerts positive and beneficial effects on HLD model rats. However, its underlying therapeutic mechanisms are not yet well understood. To explore the potential therapeutic effects of GDD against HLD, liver and urine metabolomics approach combined with histopathological examination were performed to reveal the underlying mechanisms. Changes in metabolic profiles were estimated by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) coupled with multivariate statistical analyses. The results indicated that GDD could significantly improve liver pathological variations. Moreover, 19 and 11 significantly altered metabolites were found in the liver and urine between the normal and model groups, respectively. After GDD treatment, the levels of all these disordered metabolites showed different degrees of improvement compared with the model group, including lysoPC(18:2), lysoPE(20:2/0:0), PC(18:1/14:1), alpha-linolenic acid, sphinganine, taurochenodesoxycholic acid, tetracosahexaenoic acid, 13-OxoODE, and 13-L-hydroperoxyl inoleic acid. Metabolic pathway enrichment suggested that lipid and oxidative stress metabolism were the two main pathways that participated in copper-laden rat models with GDD administration. This work indicates that GDD could achieve a therapeutic effect on HLD by ameliorating the associated metabolic disturbances.

A pilot study of oral treprostinil pharmacogenomics and treatment persistence in patients with pulmonary arterial hypertension.

BACKGROUND AND AIMS: Treprostinil is a prostacyclin analog used to treat pulmonary arterial hypertension. Dosing is empiric and based on tolerability. Adverse effects are common and can affect treatment persistence. Pharmacogenomic variants that may affect treprostinil metabolism and transport have not been well-characterized. We aimed to investigate the pharmacogenomic sources of variability in treatment persistence and dosing. METHODS: Patients were prospectively recruited from an IRB approved biobank registry at a single pulmonary hypertension center. A cohort of patients who received oral treprostinil were screened for participation. Pharmacogenomic analysis was for variants in CYP2C8, CYP2C9, and ABCC4. A retrospective review was conducted for demographics, clinical status, dosing, and response. Fisher's exact test was used for categorical data and Kruskal-Wallis test or Wilcoxon rank sum were used for continuous data. RESULTS: A total of 15 patients received oral treprostinil and were consented. Their median age was 53 years, 73% were female, and 93% were White. The median total daily dose was 22.5 mg (13.5, 41) at last clinical observation. 40% of patients discontinued treatment with a majority due to adverse effects. Approximately 27% of patients had a loss-of-function variant in CYP2C8 (*1/*3 or *1/*4), whereas 47% of patients had a loss-of-function variant in CYP2C9 (*1/*2, *1/*3, or *2/*2). Minor allele frequencies for ABCC4 (rs1751034 and rs3742106) were 0.17 and 0.43, respectively. Survival analysis showed that increased CYP2C9 activity score was associated with decreased risk for treatment discontinuation [hazard ratio (HR): 0.13; 95% confidence interval (CI): 0.02, 0.91; p = 0.04]. Genetic variants were not significantly associated with dosing. CONCLUSION: Genetic variants responsible for the metabolism and transport of oral treprostinil were common. Increased CYP2C9 activity score was associated with decreased risk for treatment discontinuation. However, dosing was not associated with genetic variants in metabolizing enzymes for treprostinil. Our findings suggest significant variability in treatment persistence to oral treprostinil, with pharmacogenomics being a potentially important contributor.The reviews of this paper are available via the supplemental material section.

Development and validation of a prediction model for axial length elongation in myopic children treated with overnight orthokeratology.

PURPOSE: To develop and validate a standardized prediction model aiming at 1-year axial length elongation and to guide the orthokeratology lens practice. METHODS: This retrospective study was based on medical records of myopic children treated with orthokeratology. Individuals aged 8-15 years (n = 1261) were included and divided into the primary cohort (n = 757) and validation cohort (n = 504). Feature selection was primarily performed to sort out influential predictors by high-throughput extraction. Then, the prediction model was developed using multivariable linear regression analysis completed by backward stepwise selection. Finally, the validation of the prediction model was performed by evaluation metrics (mean-square error, root-mean-square error, mean absolute error and R ad 2 ). RESULTS: No significant difference was found between primary and validation cohort (all p > 0.05). After the feature selection, the crude model was adjusted by demographic information in multivariable linear regression analysis, and five final predictors were identified (all p < 0.01). The interaction effect of age with 1-month change of zone-3 mm flat K was detected (p < 0.01); hence, two final prediction models were developed based on two age subgroups. The validation proved an acceptable performance. CONCLUSION: An effective multivariable prediction model aiming at 1-year axial length elongation was developed and validated. It can potentially help clinicians to predict orthokeratology efficacy and make valid adjustments. The influential variables revealed in this model can also provide designers directions to optimize the design of lens to improve the efficacy of myopia control.

Short-term effects of atropine combined with orthokeratology (ACO) on choroidal thickness.

PURPOSE: To analyse the one-month change in subfoveal choroidal thickness (SFChT) of myopic children treated with 0.01 % atropine, orthokeratology (OK), or their combination. METHODS: This is a prospective, randomized controlled trial. One hundred fifty-four children aged between 8 and 12 years with a spherical equivalent (SE) of -1.00 to -6.00 diopters were enrolled. Subjects were randomly assigned to receive 0.01 % atropine and orthokeratology (ACO, n = 39), 0.01 % atropine and single vision glasses (atropine, n = 42), orthokeratology and placebo (OK, n = 36), or placebo and single vision glasses (control, n = 37). SFChT was assessed using optical coherence tomography (OCT). Ocular parameters, including axial length (AL), were measured using a Lenstar LS 900. RESULTS: SFChT significantly increased in the ACO (14.12 ±â€¯12.88 µm, p < 0.001), OK (9.43 ±â€¯9.14 µm, p < 0.001) and atropine (5.49 ±â€¯9.38 µm, p < 0.001) groups, while it significantly decreased in the control group (-4.81 ±â€¯9.93 µm, p = 0.006). The one-month change in SFChT was significantly different between the control and treatment groups (p < 0.001). The results of pairwise comparisons among the treatment groups showed that the magnitude of the SFChT change was larger in the ACO group than in the atropine group (p = 0.002). The changes in the ACO and OK groups were not significantly different (p = 0.326). CONCLUSION: The combination of OK and atropine induced a greater increase in SFChT than monotherapy with atropine, which might indicate a better treatment effect for childhood myopia control.

Pharmacokinetic Characterization and External Evaluation of a Quantitative Framework of Sublingual Buprenorphine in Patients with an Opioid Disorder in Puerto Rico.

BACKGROUND: The aim of this analysis was to characterize the pharmacokinetics (PK) of sublingual buprenorphine (BUP) and its metabolites (buprenorphine glucuronide; BUP-g, norbuprenorphine; Nor-BUP, and norbuprenorphine glucuronide; Nor-BUP-g) in opioid use disorder (OUD) patients in Puerto Rico (PR) as a first step of evidence-based BUP dosing strategies in this population. METHODS: BUP and metabolites concentrations were measured from 0 to 8 h after the administration of sublingual buprenorphine/naloxone films in 12 stable OUD subjects. RESULTS: PK non-compartmental characteristics showed considerable variability in parameters between the subjects over the 8-h sampling time (tmax = 1.5 ± 0.7 h, Co = 1.6 ± 1.4 ng/mL, Cmax= 7.1 ± 6 ng/mL, and AUC0-8h = 26.8 ± 17.8 h·ng/mL). Subjects had a significantly higher tendency towards CYP-mediated N-demethylation, with the AUC0-8h ratios of the molar concentrations of [Nor-BUP + Nor-BUP-g] to BUP being (3.4 ± 1.9) significantly higher compared with BUP-g to BUP (0.19 ± 0.2). A two-compartment population-PK model with linear absorption (ka = 2.54 h-1), distribution (k12= 2.34 h-1, k14 = 1.29 h-1), metabolism (k24 = 1.28 × 10-1 h-1, k23 = 6.43 × 10-2 h-1, k35 = 1.23 × 10-1 h-1, k45 = 8.73 × 10-1 h-1), and elimination (k30 = 3.81 × 10-3 h-1, k50 = 1.27 × 10-1 h-1) adequately described the time-course of BUP and its metabolites, which has been externally validated using published data. CONCLUSIONS: Although limited in sampling time and number of recruited subjects, this study presents specific BUP PK characteristics that evidenced the need for additional PK studies and subsequent modeling of the data for the development of evidence-based dosing approaches in Puerto Rico.