Gene expression analysis of human induced pluripotent stem cells cryopreserved by vitrification using StemCell Keep.

In recent years, regenerative medicine research using human somatic and induced pluripotent stem cells has advanced considerably, promoting clinical applications. However, it is essential that these cells are cryopreserved safely and effectively. Most cryopreservation solution agents contain dimethyl sulfoxide (DMSO), which exhibits strong toxicity and can potentially promote cell differentiation. Hence, it is important to explore substitutes for DMSO in cryoprotectant solutions. One such alternative is StemCell Keep (SCK), a DMSO-free solution that has been reported to effectively cryopreserve human induced pluripotent stem cells (hiPS cells). To clarify the effect of cryopreservation agents on cells, DNA microarray analysis is useful, as it can identify a large number of gene expression differences in cryopreserved cells, as well as functional increases in gene groups. In this study, we performed gene expression analysis of SCK-cryopreserved hiPS cells using a DNA microarray gene chip. The hiPS cells vitrified with SCK or DMSO-based vitrification solutions were thawed and cultured on Matrigel under feeder-free conditions, and RNA was extracted for DNA microarray analysis. Genes obtained from DNA microarray data were classified by the keywords of Gene Ontology Biological Process Term, and their relationships were analyzed using DAVID or the GeneMANIA database. SCK-cryopreserved hiPS cells expressed several anti-apoptotic genes, as well as genes related to cell adhesion or proliferation at levels that were nearly equivalent to those of non-frozen hiPS cells. Gene enrichment analysis with selected genes of SCK-cryopreserved hiPS cells whose expression differences were superior to those of DAP-cryopreserved showed strong interactions of negative regulation of apoptotic process, cell adhesion and positive regulation of cell proliferation in DAVID analysis. We demonstrated that SCK successfully maintained the key functions of hiPS cells, including anti-apoptosis, cell adhesion, and cell proliferation, during cryopreservation.

D-karyo-A New Prenatal Rapid Screening Test Detecting Submicroscopic CNVs and Mosaicism.

Chromosomal microarray analysis (CMA), recently introduced following conventional cytogenetic technology, can detect submicroscopic copy-number variations (CNVs) in cases previously diagnosed as "cytogenetically benign". At present, rapid and accurate chromosomal analysis is required in prenatal diagnostics, but prenatal CMA is not widely used due to its high price and long turnaround time. We introduced a new prenatal screening method named digital karyotyping (D-karyo), which utilizes a preimplantation genetic test for the aneuploidy (PGT-A) platform. First, we conducted a preliminary experiment to compare the original PGT-A method to our modified method. Based on the preliminary results, we decided to implement the modified strategy without whole-genome amplification (WGA) and combined it with three analytical software packages. Next, we conducted a prospective study with 824 samples. According to the indication for invasive tests, the D-karyo positive rates were 2.5% and 5.0%, respectively, in the screening positive group with NT ≥ 3.5 mm and the group with fetal abnormalities by ultrasound. D-karyo is a breakthrough modality that can detect submicroscopic CNVs ≥ 1.0 Mb accurately in only 10.5 h for 24 samples at a low cost. Implementing D-karyo as a prenatal rapid screening test will reduce unnecessary CMA and achieve more accurate prenatal genetic testing than G-banding.

Increase in diastolic blood pressure induced by fragrance inhalation of grapefruit essential oil is positively correlated with muscle sympathetic nerve activity.

Fragrance inhalation of essential oils is widely used in aromatherapy, and it is known to affect blood pressure (BP) and heart rate (HR) via autonomic control of circulation. In this study, we aimed to test the hypothesis that the changes in hemodynamics with fragrance inhalation were observed along with changes in muscle sympathetic nerve activity (MSNA). In study 1, thirteen healthy men were exposed to fragrance stimulation of grapefruit essential oil for 10 min, and BP, HR, and MSNA were continuously measured. In study 2, another nine healthy men were exposed to the same fragrance stimulation; responses in BP and HR were continuously measured, and plasma noradrenaline and cortisol concentrations were determined. We found that diastolic BP increased significantly during fragrance inhalation, while the other variables remained unchanged in both studies. Although MSNA burst frequency, burst incidence, and total activity remained unchanged during fragrance inhalation, we found a significant linear correlation between changes in diastolic BP in the last 5 min of fragrance inhalation and changes in MSNA burst frequency. The plasma cortisol concentration decreased significantly at 10 min of fragrance inhalation, though the noradrenaline concentration remained unchanged. These results suggest, for the first time, that changes in BP with fragrance inhalation of essential oil are associated with changes in MSNA even with decreased stress hormone.

Aerobic Exercise Training with Brisk Walking Increases Intestinal Bacteroides in Healthy Elderly Women.

This study examined the effect of an exercise intervention on the composition of the intestinal microbiota in healthy elderly women. Thirty-two sedentary women that were aged 65 years and older participated in a 12-week, non-randomized comparative trial. The subjects were allocated to two groups receiving different exercise interventions, trunk muscle training (TM), or aerobic exercise training (AE). AE included brisk walking, i.e., at an intensity of ≥ 3 metabolic equivalents (METs). The composition of the intestinal microbiota in fecal samples was determined before and after the training period. We also assessed the daily physical activity using an accelerometer, trunk muscle strength by the modified Kraus-Weber (K-W) test, and cardiorespiratory fitness by a 6-min. walk test (6MWT). K-W test scores and distance achieved during the 6MWT (6MWD) improved in both groups. The relative abundance of intestinal Bacteroides only significantly increased in the AE group, particularly in subjects showing increases in the time spent in brisk walking. Overall, the increases in intestinal Bacteroides following the exercise intervention were associated with increases in 6MWD. In conclusion, aerobic exercise training that targets an increase of the time spent in brisk walking may increase intestinal Bacteroides in association with improved cardiorespiratory fitness in healthy elderly women.

The effects of aging on the distribution of cerebral blood flow with postural changes and mild hyperthermia.

PURPOSE: Cerebral blood flow (CBF) would be impaired with dual stresses of heat and orthostatic changes, even if those stresses are mild, in the elderly with declined cardio- and cerebrovascular functions with aging. To test the hypothesis, we compared the response of blood flow in the internal carotid artery (ICA) and vertebral artery (VA) to dual stresses of heat and orthostatic changes between the elderly and young individuals. METHODS: Nine elderly and eight young healthy men (71.3 ± 3.0 and 23.3 ± 3.1 years, mean ± SD, respectively) underwent measurements of blood flow in the ICA, VA and external carotid artery (ECA) via ultrasonography. The measurements were obtained in sitting and supine positions under normothermic (NT) and mildly hyperthermic (HT) conditions (ambient temperature 28 °C). Esophageal temperatures increased from NT (36.4 ± 0.2 °C, mean ± SE) to HT (37.4 ± 0.2 °C) with lower legs immersion in 42 °C water. RESULTS: With heat stress, ECA blood flow increased in both postures in both age groups (effect of heat, p < 0.001), whereas ICA blood flow remained unchanged. With postural changes from supine to sitting, ECA blood flow remained unchanged whereas ICA blood flow decreased (effect of posture, p = 0.027) by 18% in NT in the young and by 20% in HT in the elderly. VA blood flow remained unchanged under both heat stress and postural changes. CONCLUSIONS: The CBF is impaired under dual stresses of heat and orthostatic changes in healthy aged individuals, even if the levels of the stresses are mild.

Inhibitory Effects of Intranasal Administration of Insulin on Fat Oxidation during Exercise Are Diminished in Young Overweight Individuals.

It remains unknown whether the high insulin (INS) levels in the brain affect fat oxidation during exercise. We examined the effects of the intranasal administration of INS, which increases the INS concentration in the cerebrospinal fluid when peripheral effects are lacking, on the maximum fat oxidation rate (maxFOR) and its intensity (FATmax) during exercise in 15 young normal-weight (N group) and eight young overweight (O group) individuals. On two separate days, either INS or placebo (PL) was randomly administered intranasally before a graded exercise test. Indirect calorimetry was used to assess maxFOR and FATmax during exercise. Blood INS and glucose levels did not change after INS administration. In the N group, maxFOR and FATmax were significantly smaller in the INS trial than in the PL trial. MaxFOR was significantly smaller in the O group than in the N group and was not influenced by INS administration. Exercise-induced elevation in blood epinephrine levels tended to be reduced by INS administration only in the N group. Intranasal INS administration reduces fat oxidation during exercise without any peripheral effects, possibly by suppressing sympathetic nerve activity. This inhibitory effect is diminished in overweight subjects, suggesting that cerebral insulin effects are attenuated in this population.

Effects of 2-Year Cognitive⁻Motor Dual-Task Training on Cognitive Function and Motor Ability in Healthy Elderly People: A Pilot Study.

We aimed to examine the effect of 2-year cognitive⁻motor dual-task (DT) training on cognitive functions and motor ability of healthy elderly people without marked cognitive impairment. From the 25 participants of our 12-week DT trial conducted in 2014, we recruited 8 subjects who voluntarily participated in a new DT training program once a week for 2 years (exercise (EX) group). Their cognitive functions were evaluated by the Modified Mini-Mental State (3MS) examination and the Trail Making Test, and results were compared with those of the 11 subjects who discontinued the training and did not perform any types of exercise for 2 years (non-exercise (NO) group). Subjects in the NO group showed deterioration in the 3MS examination results, especially in the cognitive domain of attention. Meanwhile, participation in DT training maintained the scores in almost all domains of cognitive function, as well as the total 3MS scores. However, both groups had impaired quadriceps muscle strength and motor ability after the 2-year observation period. These results suggest that participating in exercise program comprising DT training for 2 years may be beneficial for maintaining the broad domains of cognitive function in healthy elderly people, although further verification is needed.

Effects of skin surface cooling before exercise on lactate accumulation in cool environment.

PURPOSE: We assessed whether plasma lactate accumulation increased and the lactate threshold (LT) declined when the skin temperature was lowered by whole body skin surface cooling before exercise in cool, but not temperate, conditions, and whether the lowered LT was associated with sympathetic activation or lowered plasma volume (PV) by cold-induced diuresis. METHODS: Ten healthy subjects performed a graded maximal cycling exercise after pre-conditioning under three different conditions for 60 min. Ambient temperature (using an artificial climatic chamber) and water temperature in a water-perfusion suit controlled at 25 and 34 °C in temperate-neutral (Temp-Neut); 25 and 10 °C in temperate-cool (Temp-Cool); and at 10 and 10 °C in cool-cool (Cool-Cool) conditions, respectively. Esophageal (Tes) and skin temperatures were measured; plasma lactate ([Lac]p) and noradrenaline concentrations ([Norad]p), and relative change in PV (%ΔPV) were determined before and after pre-conditioning and during exercise, and LT was determined. RESULTS: After pre-conditioning, Tes was not different among trials, whereas the mean skin temperature was lower in Cool-Cool and Temp-Cool than in Temp-Neut (P < 0.001). During exercise, [Lac]p and [Norad]p were higher (P = 0.009 and P < 0.001, respectively) and LT was lower (P = 0.013) in Cool-Cool than in the other trials. The %ΔPV was not different among trials. LT was correlated with [Norad]p during exercise (R = 0.50, P = 0.005). CONCLUSIONS: Whole body skin surface cooling before exercise increases lactate accumulation and decreases LT with sympathetic activation when exercise is performed in a cool, but not in a temperate, environment.

StemCell Keep™ Is Effective for Cryopreservation of Human Embryonic Stem Cells by Vitrification.

Safe and stable cryopreservation is critical for research involving human embryonic stem cells (hESCs). Dimethyl sulfoxide (DMSO) is a popular cryoprotective agent; however, its cytotoxicity cannot be ignored. Thus, there is a need for an alternate cryoprotectant. We reported previously that a novel cryopreservation reagent, StemCell Keep™ (SCK), was effective for cryopreserving human induced pluripotent stem cells (hiPSCs) by vitrification. Because hESCs and hiPSCs are not identical, the current study examined the use of SCK on hESCs. hESCs cryopreserved with SCK were thawed and cultured on SNL 76/7 cells, which were derived from a mouse fibroblast STO cell line transformed with neomycin resistance and murine LIF genes. After cryopreservation, cultured hESCs were assessed for their attachment ability and characterized by alkaline phosphatase (AP) and immunocytochemical (ICC) staining, fluorescence-activated cell sorting (FACS), reverse transcription polymerase chain reaction (RT-PCR), and karyotyping. The proliferation of SCK-cryopreserved hESCs cultured on SNL cells, or in feeder-free conditions, was higher than that of cells preserved in a solution of 2 M DMSO, 1 M acetamide, and 3 M propylene glycol (DAP). The cell number with SCK-cryopreserved hESCs was about twice that of hESCs cryopreserved in DAP. The pluripotency of SCK-cryopreserved hESCs was similar to that of DAP-cryopreserved hESCs based on AP staining. Data from ICC, FACS, and RT-PCR analyses showed that stem cell markers were continually expressed on SCK-cryopreserved hESCs. The teratoma assay showed that SCK-cryopreserved hESCs differentiated into three germ layers. Furthermore, SCK-cryopreserved hESCs had normal karyotypes. These data indicate that SCK was effective for cryopreservation of hESCs by vitrification.

Thermal sensation during mild hyperthermia is modulated by acute postural change in humans.

Thermal sensation represents the primary stimulus for behavioral and autonomic thermoregulation. We assessed whether the sensation of skin and core temperatures for the driving force of behavioral thermoregulation was modified by postural change from the supine (Sup) to sitting (Sit) during mild hyperthermia. Seventeen healthy young men underwent measurements of noticeable increase and decrease (±0.1 °C/s) of skin temperature (thresholds of warm and cold sensation on the skin, 6.25 cm2 of area) at the forearm and chest and of the whole-body warm sensation in the Sup and Sit during normothermia (NT; esophageal temperature (Tes), ∼36.6 °C) and mild hyperthermia (HT; Tes, ∼37.2 °C; lower legs immersion in 42 °C of water). The threshold for cold sensation on the skin at chest was lower during HT than NT in the Sit (P < 0.05) but not in Sup, and at the forearm was lower during HT than NT in the Sup and further in Sit (both, P < 0.05), with interactive effects of temperature (NT vs. HT) × posture (Sup vs. Sit) (chest, P = 0.08; forearm, P < 0.05). The threshold for warm sensation on the skin at both sites remained unchanged with changes in body posture or temperature. The whole-body warm sensation was higher during HT than NT in both postures and higher in the Sit than Sup during both NT and HT (all, P < 0.05). Thus, thermal sensation during mild hyperthermia is modulated by postural change from supine to sitting to sense lesser cold on the skin and more whole-body warmth.

Forced respiration during the deeper water immersion causes the greater inspiratory muscle fatigue in healthy young men.

[Purpose] The purpose of the present study was to evaluate the effect of water immersion at different water depths on respiratory function and the effect of inspiratory load breathing (ILB) during water immersion at different water depths on respiratory muscle strength evaluated by maximum inspiratory and expiratory pressures (PImax and PEmax, respectively). [Subjects] Eight healthy men participated randomly in three trials. [Methods] All sessions were conducted with the participants in a sitting position immersed in a water bath. We evaluated respiratory function, PImax and PEmax during submersion at three different levels of water depth (umbilicus; 4th-rib; or clavicle, CL) and after subsequent 15-min ILB. [Results] Decreases in vital capacity and expiratory reserve volume from baseline by water immersion were significantly greater in the CL trial than those in the other trials. In the CL trial, PImax was immediately reduced after ILB compared to that at baseline, and the reduction was significantly greater than those in the other trials. PEmax was not affected by ILB in any of the trials. [Conclusion] Forced respiration during deeper water immersion caused greater inspiratory muscle fatigue in healthy young men.

Lower thermal sensation in normothermic and mildly hyperthermic older adults.

PURPOSE: It is important to know how thermal sensation is affected by normal aging under conditions that elevate core body temperature for the prevention of heat-related illness in older people. We assessed whether thermal sensation under conditions of normothermia (NT) and mild hyperthermia (HT) is lowered in older adults. METHODS: Seventeen younger (23 ±  3 years) and 12 older (71 ±  3 years) healthy men underwent measurements of the cold and warmth detection thresholds ( ± 0.1 °C/s) of their chest and forearm skin, and whole body warmth perception under NT (esophageal temperature, T es, ~36.5 °C) and HT (T es, ~37.3 °C; lower legs immersed in 42 °C water) conditions. RESULTS: Warmth detection threshold at the forearm was increased in older compared with younger participants under both NT (P = 0.006) and HT (P = 0.004) conditions. In contrast, cold detection threshold at the forearm was decreased in older compared with younger participants under NT (P = 0.001) but not HT (P = 0.16). Mild hyperthermia decreased cold detection threshold at forearm in younger participants (P = 0.001) only. There were no effects of age and condition on warmth and cold detection thresholds at chest. Whole body warmth perception increased during HT compared with NT in both groups (both, P < 0.001), and older participants had lower values than the younger group under NT (P = 0.001) and HT (P = 0.051). CONCLUSIONS: Skin warmth detection thresholds at forearm and whole body warmth perception under NT and HT and skin cold detection thresholds at forearm under NT deteriorated with aging.

Treadmill walking in water induces greater respiratory muscle fatigue than treadmill walking on land in healthy young men.

The purpose of the present study was to investigate the effect of walking in water on respiratory muscle fatigue compared with that of walking on land at the same exercise intensity. Ten healthy males participated in 40-min treadmill walking trials on land and in water at an intensity of 60% of peak oxygen consumption. Respiratory function and respiratory muscle strength were evaluated before and after walking trials. Inspiratory muscle strength and forced expiratory volume in 1 s were significantly decreased immediately after walking in water, and expiratory muscle strength was significantly decreased immediately and 5 min after walking in water compared with the baseline. The decreases of inspiratory and expiratory muscle strength were significantly greater compared with that after walking on land. In conclusion, greater inspiratory and expiratory muscle fatigue was induced by walking in water than by walking on land at the same exercise intensity in healthy young men.

The effect of cognitive-motor dual-task training on cognitive function and plasma amyloid ß peptide 42/40 ratio in healthy elderly persons: a randomized controlled trial.

BACKGROUND: Physical activity reduces the incidence and progression of cognitive impairment. Cognitive-motor dual-task training, which requires dividing attention between cognitive tasks and exercise, may improve various cognitive domains; therefore, we examined the effect of dual-task training on the executive functions and on plasma amyloid ß peptide (Aß) 42/40 ratio, a potent biomarker of Alzheimer's disease, in healthy elderly people. METHODS: Twenty-seven sedentary elderly people participated in a 12-week randomized, controlled trial. The subjects assigned to the dual-task training (DT) group underwent a specific cognitive-motor dual-task training, and then the clinical outcomes, including cognitive functions by the Modified Mini-Mental State (3MS) examination and the Trail-Making Test (TMT), and the plasma Aß 42/40 ratio following the intervention were compared with those of the control single-task training (ST) group by unpaired t-test. RESULTS: Among 27 participants, 25 completed the study. The total scores in the 3MS examination as well as the muscular strength of quadriceps were equally improved in both groups after the training. The specific cognitive domains, "registration & recall", "attention", "verbal fluency & understanding", and "visuospatial skills" were significantly improved only in the DT group. Higher scores in "attention", "verbal fluency & understanding", and "similarities" were found in the DT group than in the ST group at post-intervention. The absolute changes in the total (8.5 ± 1.6 vs 2.4 ± 0.9, p = 0.004, 95 % confidence interval (CI) 0.75-3.39) and in the scores of "attention" (1.9 ± 0.5 vs -0.2 ± 0.4, p = 0.004, 95 % CI 2.25-9.98) were greater in the DT group than in the ST group. We found no changes in the TMT results in either group. Plasma Aß 42/40 ratio decreased in both groups following the training (ST group: 0.63 ± 0.13 to 0.16 ± 0.03, p = 0.001; DT group: 0.60 ± 0.12 to 0.25 ± 0.06, p = 0.044), although the pre- and post-intervention values were not different between the groups for either measure. CONCLUSIONS: Cognitive-motor dual-task training was more beneficial than single-task training alone in improving broader domains of cognitive functions of elderly persons, and the improvement was not directly due to modulating Aß metabolism.

Sterol-mediated regulation of human lipin 1 gene expression in hepatoblastoma cells.

Lipin 1 plays a crucial role in lipid metabolism in adipose tissue, skeletal muscle, and liver. Its physiological role involves two cellular functions: regulation of phosphatidate phosphatase activity and regulation of fatty acid oxidation. In this study, we have demonstrated that lipin 1 gene (LPIN1) expression is regulated by cellular sterols, which are key regulators of lipid metabolism. We have also characterized the sterol-response element and nuclear factor Y-binding sites in the human LPIN1 promoter. Using a luciferase assay, electrophoretic mobility shift assay, and chromatin immunoprecipitation assay, we demonstrated that these elements are responsible for the transcription of LPIN1 gene, mediated by SREBP-1 (sterol regulatory element-binding protein 1) and nuclear factor Y. Furthermore, we investigated whether lipin 1 is involved in lipogenesis by transfection of LPIN1 small interfering RNA. We infer that sterol-mediated regulation of lipin 1 gene transcription modulates triglyceride accumulation. This modulation involves changes in the activity of phosphatidate phosphatase.

Lipoprotein profiles in children with two common cholesteryl ester transfer protein gene mutations, D442G and I14A, during the first year of life.

BACKGROUND: Hyperalphalipoproteinemia is associated with cholesteryl ester transfer protein (CETP) deficiency in adults but has unclear associations in children. METHODS: We measured lipoproteins in 19 heterozygotes (D442G, n=17; I14A, n=2), one D442G/I14A compound heterozygote, 13 non-affected siblings, and 30 healthy controls at birth, 3-4 months, and 12 months. RESULTS: CETP mass was 32-70% lower in heterozygotes than in controls throughout the year. Low-density lipoprotein-cholesterol (LDL-C) was lower in heterozygotes than in controls by 30, 20, and 15% at birth, 3-4 months, and 12 months, respectively. High-density lipoprotein-cholesterol (HDL-C) was similar among the groups at birth, but was 10% higher in heterozygotes compared with controls at 3-4 and 12 months. ApoE-rich HDL-C was similar between the two groups at birth, but was 50% higher in heterozygotes than in controls at 3-4 and 12 months. These lipoprotein profile characteristics were prominent in the compound heterozygote but were not found in non-affected siblings. In heterozygotes, CETP mass correlated positively with LDL-C but negatively with HDL-C at 3-4 and 12 months. CONCLUSION: CETP is a determinant for LDL-C and HDL-C in CETP-deficient individuals in the first year of life.

Effects of bezafibrate on dyslipidemia with cholestasis in children with familial intrahepatic cholestasis-1 deficiency manifesting progressive familial intrahepatic cholestasis.

No appropriate pharmaceutical therapy has been established for dyslipidemia with cholestasis in progressive familial intrahepatic cholestasis (PFIC)-1. We evaluated the efficacy of bezafibrate in PFIC-1. We monitored the clinical presentation and lipoprotein metabolism of 3 patients, aged 3, 4, and 8 years, with FIC1 deficiency, manifesting PFIC-1, over 12 months of bezafibrate therapy. Pruritus was substantially alleviated in the 3 patients after initiation of bezafibrate. Cholestasis was alleviated in 2 of them. Serum high-density lipoprotein cholesterol and low-density lipoprotein cholesterol increased 1.6- to 2.0-fold and 1.1- to 1.2-fold, respectively; but the values remained low and normal, respectively. Serum lipoprotein X, which was at normal levels before treatment, was elevated to levels above the upper limit of the reference range. High serum triglyceride levels decreased by 15% to 30%, to normal levels, after treatment initiation. The activities of lipoprotein lipase and hepatic triglyceride lipase were increased, but those of high-density lipoprotein regulators remained unchanged. Liver expression of multidrug resistance protein-3, which regulates lipoprotein X synthesis, was enhanced by bezafibrate therapy. Bezafibrate treatment favorably affected pruritus, dyslipidemia, and cholestasis in PFIC-1.

Fluctuation of lipoprotein metabolism linked with bile acid-activated liver nuclear receptors in Alagille syndrome.

Alagille syndrome (AGS) is a rare hereditary disorder exhibiting fluctuating cholestasis and dyslipidemia. Farnesoid X receptor (FXR) and liver X receptor (LXR) are hepatic nuclear receptors that regulate bile acid and lipoprotein metabolism. To investigate whether cholestasis is related to dyslipidemia and hepatic nuclear receptor expression in AGS patients, we determined the blood levels of total bile acid (TBA) and lipoprotein parameters, and examined hepatic nuclear receptor expression in three AGS children and their three incomplete AGS parents repeatedly over several years. In the AGS children, TBA level showed significant positive correlations with low-density lipoprotein-cholesterol, apolipoprotein E (apoE)-rich high-density lipoprotein-cholesterol (HDL-C), apoA-I, apoE, and cholesteryl ester transfer protein (CETP) concentrations, but negative correlation with apoE-poor HDL-C concentration. Western blot analysis of liver biopsy specimens revealed that FXR and LXR expression increased in parallel with TBA level. CETP- and ATP-binding cassette transporter A1 expression also increased with TBA level, while scavenger receptor class B type-I expression showed the opposite response. However, apoA-I expression was similar to the control level at any TBA level. In the incomplete AGS parents, TBA and lipoprotein parameters showed little fluctuation. In summary, cholestasis is closely related to dyslipidemia and hepatic nuclear receptor expression in AGS patients.

Reduced apolipoprotein E-rich high-density lipoprotein level at birth is restored to the normal range in patients with familial hypercholesterolemia in the first year of life.

BACKGROUND: High-density lipoprotein (HDL) consists of apolipoprotein E (apoE)-rich and apoE-poor HDL particles. ApoE-rich HDL level is high at birth but decreases after birth with reciprocal elevation in low-density lipoprotein (LDL)-cholesterol. OBJECTIVES: The objective of the study was to clarify whether apoE-rich HDL decreases after birth in children with familial hypercholesterolemia (FH), a disorder caused by impaired LDL clearance. METHODS: We measured apoE-rich HDL-cholesterol and LDL-cholesterol during the first year of life in 10 FH children (one homozygote and nine heterozygotes), 12 non-FH siblings, and 75 healthy controls. RESULTS: At birth, apoE-rich HDL-cholesterol was undetectable in a homozygous FH child and lower in heterozygous FH children than non-FH siblings and controls (4+/-2 vs. 12+/-4 and 11+/-4 mg/dl, P<0.001). At 3-4 months, apoE-rich HDL-cholesterol increased in homozygous and heterozygous FH children and decreased in non-FH siblings and controls. At 12 months, apoE-rich HDL-cholesterol levels were similar among these four groups (6-7 mg/dl). In contrast, LDL-cholesterol concentration was always twice as high in heterozygous FH children as non-FH siblings and controls (at birth, 50+/-15 vs. 25+/-7 and 25+/-5 mg/dl, P<0.001; at 3-4 months of age, 159+/-29 vs. 71+/-16 and 73+/-15 mg/dl, P<0.001; at 12 months of age, 156+/-29 vs. 75+/-18 and 76+/-17 mg/dl, P<0.001). CONCLUSION: ApoE-rich HDL level is low at birth in FH children and increases to the normal level in the first year of life, opposite to the change in normal children.

Depletion of high-density lipoprotein and appearance of triglyceride-rich low-density lipoprotein in a Japanese patient with FIC1 deficiency manifesting benign recurrent intrahepatic cholestasis.

OBJECTIVE: Lipoprotein metabolism in FIC1 deficiency due to ATP8B1 mutations has never been studied sufficiently. This study was performed to investigate the detailed lipoprotein metabolism in benign recurrent intrahepatic cholestasis (BRIC) caused by FIC1 deficiency. PATIENTS AND METHODS: Lipoprotein profile and major lipoprotein regulators such as lecithin:cholesterol acyltransferase (LCAT), hepatic triglyceride lipase (HTGL), lipoprotein lipase, and cholesteryl ester transfer protein in a Japanese patient with BRIC were serially examined during a bout of cholestasis. Liver expression of farnesoid X receptor (FXR), which suppresses high-density lipoprotein (HDL) generation, was also examined. RESULTS: Hypercholesterolemia and lipoprotein X accumulation were never observed throughout this study. When the cholestasis was severe, triglyceride-rich low-density lipoprotein (LDL) accounted for most of the plasma lipoproteins whereas HDL was hardly detectable. Concurrently, activities of all regulators were decreased, together with decreases of the serum parameter for liver protein synthesis. In particular, suppressions of LCAT and HTGL activities were severe and greatly contributed to the appearance of triglyceride-rich LDL. As the cholestasis improved, this LDL gradually transformed into normal LDL with the recoveries of LCAT and HTGL activities. The activities of all regulators for the last 1 to 2 months were normal but HDL remained depleted. His liver showed low FXR expression compared with control livers. CONCLUSIONS: The present study showed an appearance of triglyceride-rich LDL due to suppressions of LCAT and HTGL activities and a depletion of HDL that is not able to be explained by lipoprotein regulators or FXR in our patient.