Sing Chung Li

Almond consumption improved glycemic control and lipid profiles in patients with type 2 diabetes mellitus.

Almond consumption is associated with ameliorations in obesity, hyperlipidemia, hypertension, and hyperglycemia. The hypothesis of this 12-week randomized crossover clinical trial was that almond consumption would improve glycemic control and decrease the risk for cardiovascular disease in 20 Chinese patients with type 2 diabetes mellitus (T2DM) (9 male, 11 female; 58 years old; body mass index, 26 kg/m²) with mild hyperlipidemia. After a 2-week run-in period, patients were assigned to either a control National Cholesterol Education Program step II diet (control diet) or an almond diet for 4 weeks, with a 2-week washout period between alternative diets. Almonds were added to the control diet to replace 20% of total daily calorie intake. Addition of approximately 60 g almonds per day increased dietary intakes of fiber, magnesium, polyunsaturated fatty acid, monounsaturated fatty acid, and vitamin E. Body fat determined with bioelectrical impedance analysis was significantly lower in patients consuming almonds (almonds vs control: 29.6% vs 30.4%). The almond diet enhanced plasma α-tocopherol level by a median 26.8% (95% confidence intervals, 15.1-36.6) compared with control diet. Furthermore, almond intake decreased total cholesterol, low-density lipoprotein cholesterol, and the ratio of low-density lipoprotein cholesterol to high-density lipoprotein cholesterol by 6.0% (1.6-9.4), 11.6% (2.8-19.1), and 9.7% (0.3-20.9), respectively. Plasma apolipoprotein (apo) B levels, apo B/apo A-1 ratio, and nonesterified fatty acid also decreased significantly by 15.6% (5.1-25.4), 17.4% (2.8-19.9), and 5.5% (3.0-14.4), respectively. Compared with subjects in the control diet, those in the almond diet had 4.1% (0.9-12.5), 0.8% (0.4-6.3), and 9.2% (4.4-13.2) lower levels of fasting insulin, fasting glucose, and homeostasis model assessment of insulin resistance index, respectively. Our results suggested that incorporation of almonds into a healthy diet has beneficial effects on adiposity, glycemic control, and the lipid profile, thereby potentially decreasing the risk for cardiovascular disease in patients with type 2 diabetes mellitus.

ENU mutagenesis identifies mice with mitochondrial branched-chain aminotransferase deficiency resembling human maple syrup urine disease

Tandem mass spectrometry was applied to detect derangements in the pathways of amino acid and fatty acid metabolism in N-ethyl-N-nitrosourea-treated (ENU-treated) mice. We identified mice with marked elevation of blood branched-chain amino acids (BCAAs), ketoaciduria, and clinical features resembling human maple syrup urine disease (MSUD), a severe genetic metabolic disorder caused by the deficiency of branched-chain alpha-keto acid dehydrogenase (BCKD) complex. However, the BCKD genes and enzyme activity were normal. Sequencing of branched-chain aminotransferase genes (Bcat) showed no mutation in the cytoplasmic isoform (Bcat-1) but revealed a homozygous splice site mutation in the mitochondrial isoform (Bcat-2). The mutation caused a deletion of exon 2, a marked decrease in Bcat-2 mRNA, and a deficiency in both BCAT-2 protein and its enzyme activity. Affected mice responded to a BCAA-restricted diet with amelioration of the clinical symptoms and normalization of the amino acid pattern. We conclude that BCAT-2 deficiency in the mouse can cause a disease that mimics human MSUD. These mice provide an important animal model for study of BCAA metabolism and its toxicity. Metabolomics-guided screening, coupled with ENU mutagenesis, is a powerful approach in uncovering novel enzyme deficiencies and recognizing important pathways of genetic metabolic disorders.

Effects of adlay bran and its ethanolic extract and residue on preneoplastic lesions of the colon in rats

BACKGROUND Adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) is a cereal crop used in traditional Chinese medicine and as a nutritious food. Epidemiologists have suspected that the low cancer rates in southeastern China might be related to adlay. Previous studies have shown that adlay has anti-tumour and anti-inflammatory activity. This study investigated the effect of adlay bran and its fractions on chemically induced colon carcinogenesis in rats. RESULTS Adlay bran and its ethanolic extract and residue significantly reduced the number of preneoplastic aberrant crypt foci (ACF) and modified their mucin composition. The inhibitory effect of adlay bran ethanolic extract on ACF showed a dose dependence. Adlay bran and its ethanolic extract suppressed small ACF (one, two or three crypts) and ACF in the distal colon, while the residue suppressed large ACF (four or more crypts). CONCLUSION These findings suggest the possibility that adlay bran and its ethanolic extract and residue inhibit colonic preneoplastic lesions in an early stage. Adlay and its fractions may have the potential to be developed as chemopreventive cereal products.

Glycogen storage disease type IV: novel mutations and molecular characterization of a heterogeneous disorder

Glycogen storage disease type IV (GSD IV; Andersen disease) is caused by a deficiency of glycogen branching enzyme (GBE), leading to excessive deposition of structurally abnormal, amylopectin-like glycogen in affected tissues. The accumulated glycogen lacks multiple branch points and thus has longer outer branches and poor solubility, causing irreversible tissue and organ damage. Although classic GSD IV presents with early onset of hepatosplenomegaly with progressive liver cirrhosis, GSD IV exhibits extensive clinical heterogeneity with respect to age at onset and variability in pattern and extent of organ and tissue involvement. With the advent of cloning and determination of the genomic structure of the human GBE gene (GBE1), molecular analysis and characterization of underlying disease-causing mutations is now possible. A variety of disease-causing mutations have been identified in the GBE1 gene in GSD IV patients, many of whom presented with diverse clinical phenotypes. Detailed biochemical and genetic analyses of three unrelated patients suspected to have GSD IV are presented here. Two novel missense mutations (p.Met495Thr and p.Pro552Leu) and a novel 1-bp deletion mutation (c.1999delA) were identified. A variety of mutations in GBE1 have been previously reported, including missense and nonsense mutations, nucleotide deletions and insertions, and donor and acceptor splice-site mutations. Mutation analysis is useful in confirming the diagnosis of GSD IV—especially when higher residual GBE enzyme activity levels are seen and enzyme analysis is not definitive—and allows for further determination of potential genotype/phenotype correlations in this disease.

Non‐lethal congenital hypotonia due to glycogen storage disease type IV

Glycogen storage disease type IV (GSD‐IV) is an autosomal recessive genetic disorder due to a deficiency in the activity of the glycogen branching enzyme (GBE). A deficiency in GBE activity results in the accumulation of glycogen with fewer branching points and long, unbranched outer chains. The disorder results in a variable phenotype, including musculoskeletal, cardiac, neurological, and hepatic involvement, alone or in continuum, which can be identified at any stage of life. The classic form of GSD‐IV is a hepatic presentation, which presents in the first 18 months of life with failure to thrive, hepatomegaly, and cirrhosis that progresses to liver failure, resulting in death by age 5 years. A severe congenital musculoskeletal phenotype with death in the neonatal period has also been described. We report an unusual case of congenital musculoskeletal presentation of GSD‐IV with stable congenital hypotonia, gross motor delay, and severe fibro‐fatty replacement of the musculature, but no hepatic or cardiac involvement. Molecular analysis revealed two novel missense mutations with amino acid changes in the GBE gene (Q236H and R262C), which may account for the mild phenotype.

Constituents in purple sweet potato leaves inhibit in vitro angiogenesis with opposite effects ex vivo

OBJECTIVE The aim of this study was to investigate the in vitro effect of polyphenols in purple sweet potato leaves (PSPLs) on angiogenesis in human umbilical vascular endothelial cells (HUVECs). The ex vivo effect was test in human serum collected from the subjects who consumed 200 g of PSPL in a low polyphenol diet versus a low polyphenol diet. METHODS Methanolic extract from PSPLs and human sera from subjects were treated with HUVECs and the effects of cell proliferation, migration, tube formation, and matrix metalloproteinase activity were investigated. RESULTS The PSPL polyphenols at 0.2 to 0.6 mM gallic acid equivalents inhibited proliferation, migration, and tube formation of vascular endothelial growth factor-treated HUVECs. Further, the activity of secreted matrix metalloproteinase-2 was decreased by at least 13.8%. However, 5% PSPL serum increased migration and tube formation of HUVECs by 110% and 56.9%, respectively, compared with serum from subjects on the low polyphenol diet. Further, the activity of matrix metalloproteinase-9 was increased by 128% in the PSPL serum. CONCLUSION These results suggest that PSPL polyphenols inhibited in vitro angiogenesis, but PSPL constituents might shift serum biochemistries to be more proangiogenic.

Riboflavin protects mice against liposaccharide-induced shock through expression of heat shock protein 25.

Riboflavin (vitamin B2) is a water-soluble vitamin essential for normal cellular functions, growth and development. This study aimed to investigate the effects of vitamin B2 on the survival rate, and expressions of tissue heat shock protein 25 (HSP25) and heat shock factor 1 (HSF1) in mice undergoing lipopolysaccharide (LPS) induced shock. Mice were assigned to four groups, saline vehicle, LPS, LPS plus low dose of vitamin B2 (LB2) and LPS plus high dose of vitamin B2 (HB2). Vitamin B2 (1 and 10mg/kg BW) was administered intraperitoneally at 2 and 0 h before the i.p. administration of LPS. At the end of the experiment, the survival rate monitored was 10, 20, 60, and 100% for LPS, LB2, HB2, and saline mice, respectively. HSP25 expressions in the heart and lung were significantly enhanced in a time-dependent manner in the HB2 mice as compared to the saline mice (p < 0.05), but not altered in the LB2 mice. In the HB2 mice, plasma riboflavin concentrations reached 300 nM at 6h post LPS and returned to the 0 h level at 72 h. The results showed that high dose of riboflavin could decrease LPS-induced mortality through an increased expression of HSP25.

Association of the congenital neuromuscular form of glycogen storage disease type IV with a large deletion and recurrent frameshift mutation.

Anderson disease, also known as glycogen storage disease type IV (MIM 232500), is a rare autosomal recessive disorder caused by a deficiency of glycogen branching enzyme. Glycogen storage disease type IV has a broad clinical spectrum ranging from a perinatal lethal form to a nonprogressive later-onset disease in adults. Here, we report 2 unrelated infants who were born small for their gestational age and who had profound hypotonia at birth and thus needed mechanical ventilation. Both of these patients shared the same frameshift mutation (c.288delA, pGly97GlufsX46) in the GBE1 gene. In addition, both of these patients were found to have 2 different large deletions in the GBE1 gene; exon 7 and exons 2 to 7, respectively, on the other alleles. This case report also highlights the need for a more comprehensive search for large deletion mutations associated with glycogen storage disease type IV, especially if routine GBE1 gene sequencing results are equivocal.

Chicken model of steroid-induced bone marrow adipogenesis using proteome analysis: a preliminary study.

BackgroundSteroid-induced adipogenesis increases fat-cell volume and pressure in bone marrow. This may be a contributing factor in some forms of osteonecrosis. In this observational study, we aimed to determine the protein expression relating to steroid-induced adipogenesis of femoral bone marrow with use of a chicken model. We compared the histologic features of the femoral marrow of eight methylprednisolone (MP)-treated chickens with those of three control chickens and assessed differential proteins with 2-dimensional gel electrophoresis and differential proteins were identified by MALDI-TOF MS.ResultsOne MP-induced chicken died of overdose anesthesia. Methylprednisolone-induced proliferation of adipose tissue and new bone formation were found on histologic examination. In our study, 13 proteins in the control and MP-induced groups were differently expressed and nine protein spots showed marked threefold downregulation after 19 weeks of MP treatment. These were serum amyloid P-component precursor, zinc finger protein 28, endothelial zinc finger protein 71, T-box transcription factor 3, cyclin-dependent kinase inhibitor 1, myosin 1D, dimethylaniline monooxygenase, and two uncharacterized proteins.ConclusionsProteomic profiling can be a useful dynamic approach for detecting protein expression in MP-induced adipogenesis of the femur in chickens.

A mutation in cartilage oligomeric matrix protein (COMP) causes early-onset osteoarthritis in a large kindred study.

We performed a genome‐wide linkage analysis to identify susceptibility loci in a large six‐generation extended family previously reported with early‐onset osteoarthritis (OA) DNA sequencing was performed to investigate involvement of the COMP (Cartilage oligomeric matrix protein) gene in this family. The region covering D19S884, D19S226, and D19S414 on chromosome 19p following genome‐wide scan from 70 individuals of this kindred showed significant linkage, with a maximum point LOD (logarithm of the odds ratio) score of 2.51 at D19S226. Direct sequencing of the COMP gene, the most plausible candidate gene in the region, identified a c.2152C>T substitution in exon 18 which resulted in a substitution of tryptophan for arginine at position 718 located in the C terminal globular domain of the gene product. A total of 26 individuals were identified with this mutation of which 21 affected individuals had the mutation, and the other five younger individuals (18.6 ± 11.3 years of age) carried the mutation without symptoms. The results indicate that COMP is the disease susceptibility gene and the c.2152C>T mutation in exon 18 could cause early‐onset OA phenotypes in this kindred, which is compatible with a previous report that this mutation also causes a mild form of multiple epiphyseal dysplasia (MED).