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Transgenic expression of DwMYB2 impairs iron transport from root to shoot in Arabidopsis thaliana.

In plant, iron uptake and homeostasis are tightly regulated to ensure its absorption from soil and to avoid excess iron in the cell. Many genes involved in this process have been identified during past several years, but there are many problems remain unsolved in the genetic regulation of whole plant iron trafficking and allocation. MYB transcription factors contain tandem repeats of a approximately 50 amino acid DNA-binding motif (R) and are involved in the regulation of many aspects of plant development, hormone signaling and metabolism. Here, we report that the ectopic expression of orchid R2R3-MYB gene DwMYB2 in Arabidopsis thaliana confers the transgenic plants hypersensitivity to iron deficiency. In DwMYB2 transgenic plants, the iron content in root is two-fold higher compared to that in wild-type root, while the reverse is true in shoot. This imbalance of iron content in root and shoot suggested that the translocation of iron from root to shoot was affected by the expression of DwMYB2 in the transgenic plants. Consistently, gene chip and reverse transcription-polymerase chain reaction analysis revealed that the ferric-chelate reductase gene, AtFRO2, and the iron transporter gene, AtIRT1 and AtIRT2, are up-regulated by DwMYB2 expression, while other potential iron transporters such as AtIREG1, AtFRD3 and NRAMP1 are down-regulated. In addition, the expression of several putative peptide transporters and transcription factors are also altered in the 35S::DwMYB2 transgenic lines. These data provide us insight into the whole plant translocation of iron and identify candidate genes for iron homeostasis in plants despite the fact that a heterologous gene was expressed.

Amino Acid Sequence↗

The mismatch DNA repair heterodimer, hMSH2/6, regulates BLM helicase.

The human MSH2/6 complex is essential for mismatch recognition during the repair of replication errors. Although mismatch repair components have been implicated in DNA homologous recombination repair, the exact function of hMSH2/6 in this pathway is unclear. Here, we show that the recombinant hMSH2/6 protein complex stimulated the ability of the Bloom's syndrome gene product, BLM, to process Holliday junctions in vitro, an activity that could also be regulated by p53. Consistent with these observations, hMSH6 colocalized with BLM and phospho-ser15-p53 in hydroxyurea-induced RAD51 nuclear foci that may correspond to the sites of presumed stalled DNA replication forks and more likely the resultant DNA double-stranded breaks. In addition, we show that hMSH2 and hMSH6 coimmunoprecipitated with BLM, p53, and RAD51. Both the number of RAD51 foci and the amount of the BLM-p53-RAD51 complex are increased in hMSH2- or hMSH6-deficient cells. These data suggest that hMSH2/6 formed a complex with BLM-p53-RAD51 in response to the damaged DNA forks during double-stranded break repair.

Adenosine Triphosphatases↗

Glypican-3-deficient mice exhibit developmental overgrowth and some of the abnormalities typical of Simpson-Golabi-Behmel syndrome.

Glypicans are a family of heparan sulfate proteoglycans that are linked to the cell surface through a glycosyl-phosphatidylinositol anchor. One member of this family, glypican-3 (Gpc3), is mutated in patients with the Simpson-Golabi-Behmel syndrome (SGBS). These patients display pre- and postnatal overgrowth, and a varying range of dysmorphisms. The clinical features of SGBS are very similar to the more extensively studied Beckwith-Wiedemann syndrome (BWS). Since BWS has been associated with biallelic expression of insulin-like growth factor II (IGF-II), it has been proposed that GPC3 is a negative regulator of IGF-II. However, there is still no biochemical evidence indicating that GPC3 plays such a role.Here, we report that GPC3-deficient mice exhibit several of the clinical features observed in SGBS patients, including developmental overgrowth, perinatal death, cystic and dyplastic kidneys, and abnormal lung development. A proportion of the mutant mice also display mandibular hypoplasia and an imperforate vagina. In the particular case of the kidney, we demonstrate that there is an early and persistent developmental abnormality of the ureteric bud/collecting system due to increased proliferation of cells in this tissue element. The degree of developmental overgrowth of the GPC3-deficient mice is similar to that of mice deficient in IGF receptor type 2 (IGF2R), a well characterized negative regulator of IGF-II. Unlike the IGF2R-deficient mice, however, the levels of IGF-II in GPC3 knockouts are similar to those of the normal littermates.

Abnormalities, Multiple↗

Cadherin-mediated adhesion is essential for myofibril continuity across the plasma membrane but not for assembly of the contractile apparatus.

The strong coordinated contraction of heart muscle is dependent on the correct alignment and connection of the myofibrils across the plasma membrane. Previous studies indicate that N-cadherin is involved in cardiac myocyte adhesion and myofibrillogenesis. To investigate whether N-cadherin is specifically required for normal myocyte structure and function, we cultured myocytes from wild-type, N-cadherin-null and mutant embryos expressing the epithelial cadherin E-cadherin. In contrast to previous studies in chicken using N-cadherin-perturbing antibodies, our in vitro studies with mouse cells demonstrate that N-cadherin is not required for myofibrillogenesis, but is critical for myofibril organization. That is, N-cadherin-deficient myocytes beat and myofibrils were well formed; however, alignment of the myofibrils through regions of cell-cell contact was lost, resulting in their random orientation. Gap junctions were perturbed in the N-cadherin-null myocytes. By contrast, focal contacts appeared normal in the mutant cells. Furthermore, E-cadherin restored normal cell morphology and behavior to the N-cadherin-deficient myocytes, including proper alignment of the myofibrils. We conclude that a different adhesive system, most probably integrin, is responsible for myofibrillogenesis in the N-cadherin-null myocytes.

Animals↗

Impaired cardiac contractile function in ventricular myocytes from leptin-deficient ob/ob obese mice.

The level of the obese gene product leptin is often positively correlated with body weight, supporting the notion that hyperleptinemia contributes to obesity-associated cardiac dysfunction. However, a link between leptin levels and cardiac function has not been elucidated. This study was designed to examine the role of leptin deficiency (resulting from a point mutation of the leptin gene) in cardiomyocyte contractile function. Mechanical properties and intracellular Ca(2+) transients were evaluated in ventricular myocytes from lean control and leptin-deficient ob/ob obese mice at 12 weeks of age. Cardiac ultrastructure was evaluated using transmission electron microscopy. ob/ob mice were overtly obese, hyperinsulinemic, hypertriglycemic, hypoleptinemic and euglycemic. Ultrastructural examination revealed swelling and disorganization of cristae in mitochondria from ob/ob mouse ventricular tissues. Cardiomyocytes from ob/ob mice displayed reduced expression of the leptin receptor Ob-R, larger cross-sectional area, decreased peak shortening and maximal velocity of shortening/relengthening, and prolonged relengthening but not shortening duration compared with lean counterparts. Consistent with mechanical characteristics, myocytes from ob/ob mice displayed reduced intracellular Ca(2+) release upon electrical stimulus associated with a slowed intracellular Ca(2+) decay rate. Interestingly, the contractile aberrations seen in ob/ob myocytes were significantly improved by in vitro leptin incubation. Contractile dysfunction was not seen in age- and gender-matched high fat-induced obese mice. These results suggested that leptin deficiency contributes to cardiac contractile dysfunction characterized by both systolic and diastolic dysfunction, impaired intracellular Ca(2+) hemostasis and ultrastructural derangement in ventricular myocytes.

Animals↗

[Effect of purified pre-beta 1 HDL on efflux of 3H-cholesterol loaded in human smooth muscle cells].

OBJECTIVE: To compare the effects of different HDL subclasses on the efflux of 3H-cholesterol loaded human smooth muscle cells (SMCS). METHODS: 3H-cholesterol loaded human SMCS were incubated with purified pre-beta 1 HDL, alpha-HDL and apoE-deficient HDL3 respectively. The cholesterol effluxing capacity and incubating time of different HDL subclasses were detected, and were analysed by double-reciprocal mapping. RESULTS: It was found that the value of Emax of pre-beta 1 HDL is higher than that of alpha-HDL and apoE-deficient HDL3 (P < 0.01), and the value of Ke of pre-beta 1 HDL is lower than that of alpha-HDL and apoE-deficient HDL3 (P < 0.01). CONCLUSION: The results showed the effluxing capacity and efficiency of pre-beta 1 HDL were greater than those of alpha-HDL and apoE-deficient HDL3, thus suggesting that pre-beta 1 HDL is a more efficient acceptor of cell-derived 3H-cholesterol.

Aorta↗

[Black diaphragm intraocular lens for congenital aniridia or traumatic iris deficiency].

OBJECTIVE: To evaluate the efficacy of a black diaphragm intraocular lens (IOL) implantation for congenital aniridia or traumatic iris deficiency and to explore the mechanism of postoperative complications. METHODS: The therapeutic effects and postoperative complications of twenty-five cases (27 eyes) of black diaphragm intraocular lens implantation were analyzed. Ultrasound biomicroscope (UBM) and gonioscopy were used to observe the position of the IOL haptics and anterior chamber angle structure in patient who developed secondary glaucoma postoperatively. RESULTS: After 3-18 months of follow-up, photophobia was reduced or disappear. The best corrected visual acuity (BCVA) Postoperation was improved 2 lines in 20 eyes, no change or within 1 line in 5 eyes, decreased 2 lines or more in 2 eyes. The complications included secondary glaucoma (10 eyes, 37.0%), astigmatism (17 eyes, 63.0%), corneal decompensation (2 eyes, 7.4%), hyphema (2 eyes, 7.4%), vitreous hemorrhage (3 eyes, 11.1%), and retinal detachment (1 eye, 3.7%). Of the 20 haptics in 10 patients, 9 were adequately located in the sulcus region, the others were located in chamber angle, ciliary processes or posterior to the ciliary processes. The changes of chamber angle included angle recession, anterior adhesion of iris and angle closure. CONCLUSIONS: The black diaphragm intraocular lens implantation provides an effective method in the treatment of congenital aniridia or traumatic iris deficiency. Chamber angle impair is major cause of postoperative secondary glaucoma. Abnormality of lens haptics position, hemorrhage and persistent inflammation were also play a role in secondary glaucoma.

Adolescent↗

[Experimental study of the evaluation of efficacy of fluid replacement based on Parkland formula for burn shock].

In order to investigate the effect of fluid resuscitation of burn shock according to Parkland formula and to find out whether it leaves any damage due to hypoxia, 22 male mongrel dogs with 50% III degrees burn were divided randomly into two groups. Group 1 (14 dogs) received no fluid replacement, whereas in group 2 (8 dogs) Ringer's lactate solution was given according to Parkland formula after the injury. Swan-Ganz catheter was inserted to the pulmonary artery before burn. FAP, CVP, PAP, cardiac output and blood gas analysis were measured preburn and at 2, 6, 12, 18, 24, 36 and 48 hours postburn. The results demonstrated that group 1 dogs all died within 12 hours postburn and group 2 dogs all survived over the shock period. However, even in these survived dogs, markedly decreased cardiac output and oxygen consumption were observed over the first 18 hours postburn. This indicated that fluid replacement according to Parkland formula can not completely correct the inadequacy of tissue perfusion, and it may leave certain damage of oxygen deficiency. Therefore, further improvement in fluid resuscitation and combined treatment for early burn shock to raise cardiac output and improve tissue oxygen delivery may be needed.

Animals↗

Differential responses of two degradation domains of HIF-1alpha to hypoxia and iron deficiency.

HIF-1alpha is a transcription factor involved in the cellular adaptation to either hypoxia or iron deficiency. In the presence of oxygen and iron, proline residues in two degradation domains are modified by HIF-1-prolyl hydroxylases (PHDs), resulting in ubiquitination and degradation of HIF-1alpha. Since both molecular oxygen and iron are elements required for this hydroxylation process, HIF-1alpha might be unmodified and stable in conditions lacking oxygen or iron. If so, two degradation domains may respond to hypoxia and iron-depletion in the same way. In this study, however, we found two degradation domains to differentially regulate the stability of HIF-1alpha. The C-terminal domain responded to both hypoxia and iron-depletion, but the N-terminal domain to only iron-depletion. The deletion or point-mutation of the C-terminal domain blunted the hypoxic induction of HIF-1alpha. However, PHD-silencing siRNAs revealed that two degradation domains were not regulated by different types of PHDs. Both domains were regulated mainly by PHD2. The further mutational analysis demonstrated that the ARD1-acetylated motif near the C-terminal degradation domain (CDD) modulates the oxygen-dependent regulation of HIF-1alpha. The oxygen-dependent HIF-1alpha regulation requiring both proline hydroxylation and lysine acetylation may be more complicated than the iron-dependent regulation requiring only proline hydroxylation.

Cell Line↗

Alpha1-antitrypsin and neutrophil elastase imbalance and lung cancer risk.

OBJECTIVE: Imbalance between alpha(1)-antitrypsin and neutrophil elastase is an underlying cause of lung tissue damage that may create a favorable host environment for carcinogenesis. We conducted a case-control study to investigate whether genetic variations indicative of alpha(1)-antitrypsin deficiency (A1ATD) or an excess of neutrophil elastase modify lung cancer risk DESIGN: The case patients were 305 consecutively identified primary lung cancer patients, and the control subjects were 338 community residents. Protease inhibitor-1 (PI1), encoding alpha(1)-antitrypsin, was typed by an isoelectric focusing assay. Neutrophil elastase-2 (ELA2), encoding neutrophil elastase, was typed by two single-nucleotide polymorphism sites. Multivariable logistic regression models tested the independent and interactive effects of PI1, ELA2, tobacco smoke exposure, COPD, and family history of lung cancer RESULTS: Sex and ethnicity were comparable between case patients and control subjects, but case patients were more likely to be smokers, and to have a history of COPD, environmental tobacco smoke exposure, and a positive family history of lung cancer. Haplotype analysis indicated an overall strong association between the two ELA2 markers and lung cancer risk. Our best-fitting model showed significant and independent effects of the PI1-deficient allele (odds ratio [OR], 2.0; 95% confidence interval [CI], 1.4 to 3.0) and the ELA2 T-G haplotype (OR, 4.1; 95% CI, 1.9 to 8.9) on lung cancer risk, and an increased risk (OR, 2.6; 95% CI, 2.4 to 2.8) for individuals carrying both a PI1-deficient allele and a G-G haplotype CONCLUSIONS: Genotypes indicative of A1ATD and/or an excess of neutrophil elastase are significantly associated with lung cancer risk. Our findings may provide opportunities to better understand the mechanisms of lung cancer development and risk reduction.

Aged↗

[A description on the Chinese national nutrition and health survey in 2002].

OBJECTIVE: To describe the status of nutrition and health related indices in the Chinese population. METHODS: A stratified multi-stage cluster sampling was used from 31 provinces, autonomous regions, and municipalities. The survey was done in 2002, including data gathered from questionnaires, interviews, physical examinations, measurement of biochemical indices, and dietary investigation. RESULTS: Cereals accounted for 48.5% of all the sources of energy in urban and 61.4% in rural populations. Daily mean percentages of calories for total fat were 35.0% in urban and 27.5% in rural areas. The prevalence rates of stunting and underweight were 14.3% and 7.8% respectively in young children under 5-year-old. The prevalence of vitamin A deficiency was 9.3% in Chinese children aged 3-12 years old. The total prevalence of anemia was 15.2% in general population of all ages. The prevalence of anemia in young adults was significantly higher in women than in men. The total prevalence rates of overweight and obesity were 17.6% and 5.6%, respectively. The prevalence rates of hypertension, diabetes, hypercholesteremia, hypertriglyceridemia, or low serum high density lipoprotein cholesterol were 18.8%, 2.6%, 2.9%, 11.9%, 7.4% respectively in Chinese adults aged 18 and over. The rates of awareness, treatment, and under control among hypertensives were 30.2%, 24.7%, and 25.0%, respectively. Significant regional and age differences were revealed in the dietary habit and the prevalence of various diseases. The prevalence of diseases associated with malnutrition were higher in rural than in urban areas. In contrast, the prevalence of conditions associated with overconsumption and inappropriate dietary patterns were higher in urban than in rural populations. CONCLUSION: Chinese people were currently suffering from both problems on nutrition related issues and burdens of diseases which were characterized in nutrient deficiencies and overconsumption, malnutrition and noncommunicable conditions associated with overconsumption and inappropriate diet. The difference of nutrition and health status between rural and urban people was also seen.

Adolescent↗

The clinicopathological correlation of epithelial subtyping in thymoma: a study of 112 consecutive cases.

The clinicopathological features of 112 thymomas collected from the surgical pathological files of Taipei Veterans General Hospital from 1961 to 1991 were investigated to determinate the clinical efficacy of epithelial subtyping. All thymomas were categorized based on the Müller-Hermelink system into three subtypes: cortical thymoma, mixed thymoma, and medullary thymoma. The former was further subclassified into organoid thymoma, conventional cortical thymoma, and well differentiated thymic carcinoma (WDTC) according to the systems of Pescarmona and Kirchner. The association of each subtype with sex, age at diagnosis, clinical stage, presence of myasthenia gravis, and length of survival was studied. As classified by the Müller-Hermelink system, the cortical thymomas as a whole tended to occur in younger patients and were more frequently associated with myasthenia gravis than the medullary thymomas. The cortical thymomas also showed a propensity to be invasive in nature, whereas the medullary thymomas generally behaved as benign tumors. Further subclassification of cortical thymomas into organoid thymoma, conventional thymoma, and WDTC did not provide more information about clinical behavior. By Kaplan-Meier's actuarial survival analyses none of the epithelial subtypes displayed a statistically significant influence on prognosis. It is concluded that staging remains the most important factor affecting the patient's outcome. Because of the existence of many intermediate forms and the deficiency of clinical relevance, the subclassification of cortical thymomas should be interpreted as a morphological continuum rather than as distinct histological variants.

Adolescent↗

Developmental expression of PAM (protein associated with MYC) in the rodent brain.

Recently, human protein associated with MYC, PAM, has been cloned and characterized as a large protein that interacts with the transcriptional-activating domain of Myc. The regional expression pattern of PAM in brains has not been yet been defined. Expression patterns of PAM in both rat and mouse brains were examined by using in situ hybridization. Here, we demonstrate that PAM mRNA is highly expressed in specific anatomical regions including hippocampus, dentate gyrus and cerebellum. In these areas, PAM mRNA is restricted to pyramidal cells of hippocampus and granule cells of dentate gyrus and cerebellum. During development, PAM mRNA expression is differentially regulated. It is turned on after birth and up-regulated during the first postnatal 2 weeks. Thereafter, PAM mRNA expression remains elevated into adulthood. The regional distribution of PAM in brain is similar to that observed for several adenylyl cyclase isoforms such as type I isoform. However, no obvious alterations of PAM mRNA expression are detected in brains of mice deficient in type I or type 8 or type 1 and type 8 isoforms of adenylyl cyclase. Thus, adenylyl cyclase does not appear to alter the expression of PAM.

Adaptor Proteins, Signal Transducing↗

In vitro and in vivo studies of MutS, MutL and MutH mutants: correlation of mismatch repair and DNA recombination.

We have used the recently determined crystal structures of Escherichia coli (E. coli) MutS, MutL and MutH to guide construction of 47 amino-acid substitutions in these proteins and analyzed their behavior in mismatch repair and recombination in vitro and in vivo. We find that the active site of the MutH endonuclease is composed of regions from two separate structural domains and that the C-terminal 5 residues of MutH influence both DNA binding and cleavage. We also find that the non-specific DNA-binding activity of MutL is required for mismatch repair and probably functions after strand cleavage by MutH. Alteration of residues in either the mismatch recognition domain, the ATPase active site, or the domain interfaces linking the two activities can diminish the differential binding of MutS to homoduplex versus heteroduplex and results in the loss of mismatch-specific MutH activation. Finally, every mutation that abolishes mismatch repair is deficient in blocking homeologous recombination, suggesting that mismatch repair and prevention of homeologous recombination use the same MutS-MutL complexes for signaling in E. coli.

Adenosine Triphosphatases↗

Propionyl coenzyme A carboxylase deficiency presenting as non-ketotic hyperglycinaemia.

A 4-month-old girl presented with myoclonic seizures and an electroencephalogram showing hypsarrhythmia. Hyperglycinuria and a cerebrospinal fluid to plasma glycine ratio of 0.2 suggested the diagnosis of non-ketotic hyperglycinaemia. Propionic acid and methyl citric acid were present in the urine, and propionyl coenzyme A carboxylase was deficient in leucocytes and fibroblasts. The ketotic and non-ketotic hyperglycinaemias cannot be differentiated by CSF: plasma glycine ratios.

Carboxy-Lyases↗

Repeated alpha-galactosylceramide administration results in expansion of NK T cells and alleviates inflammatory dermatitis in MRL-lpr/lpr mice.

NK T (NKT) cells expressing the invariant Valpha14-Jalpha18 TCR alpha-chain recognize glycolipid Ags such as alpha-galactosylceramide (alpha-GalCer) presented by the MHC class I-like molecule CD1d. Upon activation by alpha-GalCer, invariant NKT cells secrete multiple cytokines and confer protection in certain immune-mediated disorders. Here we have investigated the role of NKT cells in the development of inflammatory dermatitis in MRL-lpr/lpr mice, which shares features with lupus in humans. Our results show that the numbers Sand functions of NKT (TCRbeta(+)CD1d/alpha-GalCer tetramer(+)) cells, particularly of the NK1.1(-) subset, are reduced in MRL-lpr/lpr mice compared with MRL-fas/fas and/or nonautoimmune C3H/Hej and BALB/c mice. Repeated treatments with alpha-GalCer result in the expansion of NKT cells and alleviate dermatitis in MRL-lpr/lpr mice. Our results indicate that NKT cell deficiency can be corrected by repeated alpha-GalCer treatment and that NKT cells may play a protective role in inflammatory dermatitis of lupus-prone mice.

Animals↗

Clinical features of type 1 diabetic children at initial diagnosis.

Clinical and laboratory data of 165 newly diagnosed diabetic children at Chang Gung Children's Hospital in Taiwan from 1993 to 2003 were analyzed. The age distribution was categorized as 0-1 years: 19 (11.5%), 2-4 years: 39 (23.6%), 5-9 years: 61 (37%), and 10-16 years: 46 (27.9%); the incidence showed spring/winter peak and a mid-summer trough but not statistically significant (chi2= 1.86, p=0.61). The most common symptoms on presentation were polydipsia, weight loss, polyuria, and fatigue. Very young diabetic children had a higher incidence of having dyspnea at presentation (p=0.029) and had significantly lower levels of C-peptide (p=0.0086) and HbA1c (p=0.0025). The prevalences of GAD antibody and insulin autoantibodies were 67% and 49%, respectively. Children with severe diabetic ketoacidosis (DKA) were younger at diabetes onset (p=0.36, not to the statistical significance), had a higher incidence of preceding febrile illnesses (p=0.05), lower T4 and T3 levels (p=0.0008, 0.02), and higher white blood cell counts (p=0.03) than the mild group. In conclusion, diabetes in very young children is not rare and is more associated with more severe insulin deficiency. Young age at diabetes onset and preceding febrile illness may provoke severe DKA. High alertness is necessary for prompt diagnosis and management.

Blood Glucose↗