Miracle-dz/UniFAR-FTUs huggingface dataset:
Fukutin gene retrotransposal insertion in a non‐Japanese Fukuyama congenital muscular dystrophy (FCMD) patient

Fukuyama‐type congenital muscular dystrophy (FCMD) is an autosomal recessive disorder, characterized by severe muscular dystrophy associated with brain malformation. FCMD is the second most common form of muscular dystrophy and one of the most common autosomal recessive diseases among the Japanese population; however, no typical FCMD cases have been reported in any other population. In this study, we report on the first identification of a Chinese FCMD patient; our findings are supported by clinical, histological, and magnetic resonance imaging (MRI) evidence, as well as fukutin gene mutational analyses. The patient presented with neonatal hypotonia, seizures, and delayed motor and speech development. Additional testing revealed cerebral and cerebellar gyrus abnormalities with white matter signal intensity changes, elevated serum creatine kinase (CK) levels, and dystrophic skeletal muscle with α‐dystroglycan hypoglycosylation, and normal β‐dystroglycan and merosin expression. Genetic analysis of the fukutin gene showed one copy with a Japanese founder 3‐kilobase (kb) retrotransposal insertion in the 3′‐non‐coding region and the other copy with a known c.139C>T mutation. This is the first FCMD case reported in the Chinese population and the first case in which the 3‐kb insertion has been found outside of the Japanese population. This report emphasizes the importance of considering the fukutin founder mutation for diagnostic purposes outside of Japan. © 2009 Wiley‐Liss, Inc.
Screening of DNA methylation at the H19 promoter or the distal region of its ICR1 ensures efficient detection of chromosome 11p15 epimutations in Russell–Silver syndrome

Over a 10‐year period blood samples were collected from 57 individuals with growth restriction and RSS‐like features. Our goal was to identify epigenetic abnormalities in this cohort, including uniparental disomy of chromosome 7 (UPD7), methylation changes at chromosome11p15, as well as new epigenomic alterations. We evaluated the methylation status of 7 imprinting control regions on chromosomes 7, 11, 14, and 15. UPD7 and chromosome 7 structural abnormalities had been previously identified in five patients. Epigenetic alterations on chromosome 11p15 were identified in 11 patients. Of interest, in 3 of these 11 patients, the epigenetic alterations were limited to the H19 promoter and the distal region of its associated imprinting center, ICR1. In addition, in one patient, we detected methylation changes consistent with maternal UPD at all tested imprinted regions. This patient series suggests that epimutations on chromosome 11p15 can be most efficiently detected in RSS patients by screening for DNA methylation defects at the H19 promoter or the distal region of ICR. © 2009 Wiley‐Liss, Inc.
A Complex of Nuclear Factor I-X3 and STAT3 Regulates Astrocyte and Glioma Migration through the Secreted Glycoprotein YKL-40*

Nuclear factor I-X3 (NFI-X3) is a newly identified splice variant of NFI-X that regulates expression of several astrocyte-specific markers, such as glial fibrillary acidic protein. Here, we identified a set of genes regulated by NFI-X3 that includes a gene encoding a secreted glycoprotein YKL-40. Although YKL-40 expression is up-regulated in glioblastoma multiforme, its regulation and functions in nontransformed cells of the central nervous system are widely unexplored. We find that expression of YKL-40 is activated during brain development and also differentiation of neural progenitors into astrocytes in vitro. Furthermore, YKL-40 is a migration factor for primary astrocytes, and its expression is controlled by both NFI-X3 and STAT3, which are known regulators of gliogenesis. Knockdown of NFI-X3 and STAT3 significantly reduced YKL-40 expression in astrocytes, whereas overexpression of NFI-X3 dramatically enhanced YKL-40 expression in glioma cells. Activation of STAT3 by oncostatin M induced YKL-40 expression in astrocytes, whereas expression of a dominant-negative STAT3 had a suppressive effect. Mechanistically, NFI-X3 and STAT3 form a complex that binds to weak regulatory elements in the YKL-40 promoter and activates transcription. We propose that NFI-X3 and STAT3 control the migration of differentiating astrocytes as well as migration and invasion of glioma cells via regulating YKL-40 expression.
Relationship of impairment induced by intracellular S‐adenosylhomocysteine accumulation with DNA methylation in human umbilical vein endothelial cells treated with 3‐deazaadenosine

The aim of this study was to estimate the relationship of endothelial dysfunction induced by intracellular S‐adenosylhomocysteine (SAH) accumulation and DNA methylation in human umbilical vein endothelial cells (HUVEC). The isolated HUVEC were incubated with 3‐deazaadenosine (DZA) to induce experimental intracellular SAH accumulation. The impairment of HUVEC function was assessed by changes in morphology and proliferative ability. The expression of DNA methyltransferase‐1 (DNMT1) and the atherosclerosis related genes [oestrogen receptor‐alpha (ER‐α), extracellular superoxide dismutase (EC‐SOD) and monocyte chemoattractant protein‐1 (MCP‐1)] were analysed using quantitative real‐time PCR. Global DNA methylated status was measured using the cytosine extension assay. The methylated patterns of ER‐α, EC‐SOD and MCP‐1 genes were determined with methylation‐specific PCR. We found that DZA administration increased intracellular SAH levels progressively and simultaneously decreased Hcy content in medium. Moreover, the supplementation induced HUVEC apoptosis, inhibited proliferation ability and DNMT1 mRNA expression (P < 0.05) and furthermore reduced global DNA methylation status (P < 0.05). Correlation analysis showed the presence of a negative correlation between intracellular SAH concentration, proliferative ability, and expression of ER‐α, EC‐SOD, and DNMT1 (r = −0.89, −0.86, −0.92 and −0.88 respectively, P < 0.001); and a positive correlation with MCP‐1 expression and DNA [3H]‐dCTP incorporation (r = 0.89 and 0.93 respectively, P < 0.001). Our results showed that endothelial dysfunction induced by intracellular SAH accumulation is mediated by regulating the expression of atherosclerosis related genes in HUVEC, which is not related with gene promoter methylated patterns, but may be associated with altered global DNA hypomethylated status. These findings suggest that SAH can act as the potential molecular biological marker in the promotion of atherogenesis.
Universities and medical education in Nigeria

Formal attempts at Medical Education in Nigeria began in 1927 with the establishment of an institution in Lagos for training medical manpower to diploma level. They were trained to practice only in Nigeria. The program was not popular and was discontinued. Following the report of the Elliot Commissions on higher education in West Africa it was decided to establish the University of London College at Ibadan, with a Faculty of Medicine as one of the initial faculties. This was realized in 1948. The debate on what type of doctor to produce for Nigeria ended with the decision to produce high caliber doctors of the same standing as British trained doctors. In 1960 the Ashby Commission on Higher Education in Nigeria recommended the establishment of more training institutions, including those for medicine. This led to the establishment of the University of Lagos with the College of Medicine. The three initial regional governments all established their universities with medical faculties. Medical education has expanded rapidly with the expansion of universities, and we now have Federal and State governments as well as other organizations or private individuals owning universities with medical schools. Regulation of undergraduate medical education has continued to be under the dual oversight of the National Universities Commission and the Medical and Dental Council of Nigeria. The main problems of the medical schools have been the shortage of properly trained staff and poor facilities, curriculum stagnation and lack of modern teaching and assessment instruments. To tackle these problems training in educational methods should be mandatory for academic staff; there should be greater synergy between the NUC and MDCN, and curriculums should be reviewed to reflect modern trends.
Effects of Trypanosoma cruzi infection on myocardial morphology, single cardiomyocyte contractile function and exercise tolerance in rats

The aim of this study was to investigate the effects of Trypanosoma cruzi (T. cruzi) infection on myocardial morphology, single cardiomyocyte contractile function and exercise tolerance in rats. Adult Wistar rats were randomized into control (n = 14) and infected (n = 14) groups. Infected animals were inoculated with T. cruzi Y strain (300,000 trypomastigotes/50 g body weight). After 9 weeks, the animals were subjected to a treadmill running protocol. Then, the right atrium (RA) and left ventricle (LV) were removed for morphological and cell contractile evaluation. The infected animals exhibited a significant reduction in distance travelled, total time to fatigue and workload. In addition, these animals had hypertrophy, increased myocardial cellularity, and an increase in the proportion of collagen and blood vessels. RA and LV myocytes from infected animals showed marked contractile dysfunction under basal conditions and a reduced contractile response to β‐adrenergic stimulation. The workload of infected animals was correlated closely with the amplitude of cell shortening of RA and LV myocytes. T. cruzi infection influenced the myocardial morphology and the mechanical properties of RA and LV single myocytes negatively and reduced exercise tolerance. Single cardiomyocyte contractile dysfunction could constitute an additional mechanism of cardiac impairment and reduced exercise tolerance in this infection.
Transforming growth factor‐β enhances invasion and metastasis in Ras‐transfected human malignant epidermal keratinocytes

Transforming growth factor‐β (TGF‐β) is known to act as a tumour suppressor early in carcinogenesis, but then switches to a pro‐metastatic factor in some late stage cancers. However, the actions of TGF‐β are context dependent, and it is currently unclear how TGF‐β influences the progression of human squamous cell carcinoma (SCC). This study examined the effect of overexpression of TGF‐β1 or TGF‐β2 in Ras‐transfected human malignant epidermal keratinocytes that represent the early stages of human SCC. In vitro, the proliferation of cells overexpressing TGF‐β1 or TGF‐β2 was inhibited by exogenous TGF‐β1; cells overexpressing TGF‐β1 also grew more slowly than controls, but the growth rate of TGF‐β2 overexpressing cells was unaltered. However, cells that overexpressed either TGF‐β1 or TGF‐β2 were markedly more invasive than controls in an organotypic model of SCC. The proliferation of the invading TGF‐β1 overexpressing cells in the organotypic assays was higher than controls. Similarly, tumours formed by the TGF‐β1 overexpressing cells following transplantation to athymic mice were larger than tumours formed by control cells and proliferated at a higher rate. Our results demonstrate that elevated expression of either TGF‐β1 or TGF‐β2 in cells that represent the early stages in the development of human SCC results in a more aggressive phenotype.
HIV-1 Tat Protein Promotes Neuronal Dysfunction through Disruption of MicroRNAs*

Over the last decade, small noncoding RNA molecules such as microRNAs (miRNAs) have emerged as critical regulators in the expression and function of eukaryotic genomes. It has been suggested that viral infections and neurological disease outcome may also be shaped by the influence of small RNAs. This has prompted us to suggest that HIV infection alters the endogenous miRNA expression patterns, thereby contributing to neuronal deregulation and AIDS dementia. Therefore, using primary cultures and neuronal cell lines, we examined the impact of a viral protein (HIV-1 Tat) on the expression of miRNAs due to its characteristic features such as release from the infected cells and taken up by noninfected cells. Using microRNA array assay, we demonstrated that Tat deregulates the levels of several miRNAs. Interestingly, miR-34a was among the most highly induced miRNAs in Tat-treated neurons. Tat also decreases the levels of miR-34a target genes such as CREB protein as shown by real time PCR. The effect of Tat was neutralized in the presence of anti-miR-34a. Using in situ hybridization assay, we found that the levels of miR-34a increase in Tat transgenic mice when compared with the parental mice. Therefore, we conclude that deregulation of neuronal functions by HIV-1 Tat protein is miRNA-dependent.
Oral Streptococcus species in pre-term and full-term children - a longitudinal study.

BACKGROUND Despite high clinical significance, the microbiology of the dental biofilm in young children remains poorly understood. AIM The aim of this longitudinal study was to investigate five Streptoccocus species commonly found in the oral biofilm of children, namely Streptococcus mutans, Streptococcus sobrinus, Streptococcus mitis, Streptococcus sanguinis, and Streptococcus salivarius to determine their relative numbers in caries-free pre-term children, and age-matched full-term controls. DESIGN Plaque and saliva samples were obtained from 15 pre-term children and 15 age-matched controls at ages 3, 6, 12, 18, and 24 months. A quantitative real-time PCR technique was used to determine the numbers of five species of Streptococcus using probes and primers specific for each bacterial species. RESULTS All species of Streptococcus generally increased from ages 3 to 24 months. The relative ratios of the bacteria remained fairly constant at all ages studied (P > 0.1). There were no significant differences in numbers of all Streptococcus species between pre-term children and full-term controls at all the ages investigated between. CONCLUSION The results show that the relative numbers of S. mutans, S. sobrinus, S. mitis, S. sanguinis, and S. salivarius remain relatively constant from 3 to 24 months of age in caries-free pre- and full-term children.
Characterization of enamel in primary teeth by optical coherence tomography for assessment of dental caries.

BACKGROUND Caries is a disease that affects both primary and permanent dentitions, therefore new methods of caries diagnosis need to be tested on primary teeth as well as on permanent teeth. AIM This study reports the application of optical coherence tomography (OCT) to characterize sound dental structure and detect natural caries of human primary teeth. DESIGN Six primary teeth were sectioned into thin slices ( approximately 1.5 mm), and analysed perpendicular to the enamel surface by two home-made OCT systems operating around 1280 and 840 nm. The generated images were compared with histology as the gold standard. RESULTS We demonstrated the efficacy of the OCT technique to measure the depth of the enamel layer, whose result was statistically compared with histology (P = 0.823; P > 0.05) with good agreement. We also determined, through measurement of contrast values, an increase in backscattered intensity of the order of two to three times between sound and caries regions. CONCLUSIONS We employed OCT generated images to characterize the enamel layer. The technique showed great potential to be used on paediatric dentistry clinical on early caries detection with no pain, as it is a noninvasive method.
Folding Delay and Structural Perturbations Caused by Type IV Collagen Natural Interruptions and Nearby Gly Missense Mutations*

Background: Nonfibrillar collagens contain natural interruptions in the (Gly-Xaa-Yaa)n sequence pattern. Results: An interruption within a recombinant triple helix led to delayed folding, an effect magnified by a nearby Gly missense mutation. Conclusion: The length and relative positions of interruptions influence triple-helix folding. Significance: Consequences of natural interruptions can be compared with pathogenic mutations to clarify the basis of collagen disorders. The standard collagen triple helix requires Gly as every third residue in the amino acid sequence, yet all nonfibrillar collagens contain sites where this repeating pattern is interrupted. To explore the effects of such natural interruptions on the triple helix, a 4- or 15-residue sequence from human basement membrane type IV collagen was introduced between (Gly-Xaa-Yaa)n domains within a recombinant bacterial collagen. The interruptions had little effect on melting temperature, consistent with the high thermal stability reported for nonfibrillar collagens. Although the 4-residue interruption cannot be accommodated within a standard triple helix, trypsin and thermolysin resistance indicated a tightly packed structure. Central residues of the 15-residue interruption were protease-susceptible, whereas residues near the (Gly-Xaa-Yaa)n boundary were resistant, supporting a transition from an alternate conformation to a well packed triple helix. Both interruptions led to a delay in triple-helix folding, with the 15-residue interruption causing slower folding than the 4-residue interruption. These results suggest that propagation through interruptions represents a slow folding step. To clarify the relation between natural interruptions and pathological mutations, a Gly to Ser missense mutation was placed three triplets away from the 4-residue interruption. As a result of this mutation, the 4-residue interruption and nearby triple helix became susceptible to protease digestion, and an additional folding delay was observed. Because Gly missense mutations that cause disease are often located near natural interruptions, structural and folding perturbations arising from such proximity could be a factor in collagen genetic diseases.
X‐linked hereditary hemihypotrophy hemiparesis hemiathetosis

Hereditary hemihypotrophy hemiparesis hemiathetosis syndrome (HHHH; OMIM 306960) was first reported in a family with congenital left hemiplegia in two males and moderately affected females. We describe a family with three males demonstrating congenital right hemiplegia with porencephalic lesions of the left internal capsule and putamen, or the periventricular white matter just above the internal capsule. Clinical findings within each family are limited to the same side, though sidedness differed between families. Both pedigrees were most consistent with X‐linked inheritance. Genome‐wide linkage analysis in our family further supports a locus for HHHH on chromosome X. © 2010 Wiley‐Liss, Inc.
Position Paper on Critical Care Pharmacy Services

Objective. To identify and describe the scope of practice that characterizes the critical care pharmacist and critical care pharmacy services. Specifically, the goals were to define the level of clinical practice and specialized skills characterizing the critical care pharmacist as clinician, educator, researcher, and manager; and to recommend fundamental, desirable, and optimal pharmacy services and personnel requirements for the provision of pharmaceutical care to critically ill patients. Hospitals having comprehensive resources as well as those with more limited resources were considered.
LRP130 Protein Remodels Mitochondria and Stimulates Fatty Acid Oxidation*

Background: Impaired oxidative phosphorylation (OXPHOS) is implicated in several metabolic disorders. Hence, molecules that stimulate OXPHOS may prove beneficial. Results: LRP130, a protein involved in Leigh syndrome, activates mitochondrial transcription, which stimulates OXPHOS and oxidative metabolism in cells and mouse liver. Conclusion: By activating mitochondrial transcription, LRP130 remodels mitochondria resulting in denser cristae. Significance: An activator of OXPHOS, LRP130 may mitigate certain metabolic disorders. Impaired oxidative phosphorylation (OXPHOS) is implicated in several metabolic disorders. Even though mitochondrial DNA encodes several subunits critical for OXPHOS, the metabolic consequence of activating mitochondrial transcription remains unclear. We show here that LRP130, a protein involved in Leigh syndrome, increases hepatic β-fatty acid oxidation. Using convergent genetic and biochemical approaches, we demonstrate LRP130 complexes with the mitochondrial RNA polymerase to activate mitochondrial transcription. Activation of mitochondrial transcription is associated with increased OXPHOS activity, increased supercomplexes, and denser cristae, independent of mitochondrial biogenesis. Consistent with increased oxidative phosphorylation, ATP levels are increased in both cells and mouse liver, whereas coupled respiration is increased in cells. We propose activation of mitochondrial transcription remodels mitochondria and enhances oxidative metabolism.
Myocardin-related Transcription Factor-A Complexes Activate Type I Collagen Expression in Lung Fibroblasts*

Background: Myofibroblasts are responsible for excessive collagen expression during scarring and fibrosis. Results: Myocardin-related transcription factor-A (MRTF-A) potentiates collagen overproduction through transcription complexes containing Sp1 and SRF on noncanonical DNA elements. Conclusion: MRTF-A promotes myofibroblast differentiation and collagen production through diverse molecular mechanisms. Significance: MRTF-A may be a molecular target in developing strategies to impede fibrosis. Pulmonary fibrosis is characterized by the excessive deposition of a collagen-rich extracellular matrix. The accumulation of collagen within the lung interstitium leads to impaired respiratory function. Furthermore, smooth muscle actin-positive myofibroblasts within the fibrotic lung contribute to disease progression. Because collagen and smooth muscle cell α-actin are coordinately expressed in the setting of fibrosis, the hypothesis was tested that specific transcriptional regulators of the myocardin family might also regulate collagen gene expression in myofibroblasts. Myocardin-related transcription factors (MRTFs), through their interaction with the serum-response factor (SRF) on CArG box regulatory elements (CC(A/T)6GG), are important regulators of myofibroblast differentiation. MRTF-A transactivated type I collagen gene reporters as much as 100-fold in lung myofibroblasts. Loss of functional MRTF-A using either a dominant negative MRTF-A isoform, shRNA targeting MRTF-A, or genetic deletion of MRTF-A in lung fibroblasts significantly disrupted type I collagen synthesis relative to controls. Analysis of the COL1A2 proximal promoter revealed a noncanonical CArG box (CCAAACTTGG), flanked by several Sp1 sites important for MRTF-A activation. Chromatin immunoprecipitation experiments confirmed the co-localization of MRTF-A, SRF, and Sp1 bound to the same region of the COL1A2 promoter. Mutagenesis of either the noncanonical CArG box or the Sp1 sites significantly disrupted MRTF-A activation of COL1A2. Together, our findings show that MRTF-A is an important regulator of collagen synthesis in lung fibroblasts and exhibits a dependence on both SRF and Sp1 function to enhance collagen expression.