bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expanding the Use of Tyrosine Phenol Lyase in the Enzymatic Synthesis of Unnatural Amino Acids
Authors: Munozvilla Jubilee H. , Loo Christian E. , Schwans Jason P. 
Published: April 2020

The use of enzymes in the synthesis of pharmaceuticals and biochemical tools offers an attractive approach to generating complex structures, as enzymatic reactions often lead to the formation of specific products in high yields. Tyrosine phenol lyase (TPL), an enzyme biologically involved in the degradation of tyrosine, has been used to synthesize a variety of substituted tyrosines including fluorotyrosines for biochemical studies. In addition to fluorotyrosines, expanding the scope of substrates to naphthol‐based compounds offers an attractive target for enzymatic synthesis of complex molecules, as substituted naphthols can be used as biochemical probes and as building blocks in potential pharmaceuticals. Using Citrobacter freundii TPL we enzymatically synthesized the naphthol‐analog of tyrosine, but the unnatural amino acid was generated in low yield. The simplest model for the low conversion is that the larger substrate is poorly accommodated by the enzyme active site and this poor binding leads to the low yield in the enzymatic reaction. To test this hypothesis, we mutated several bulky residues near the substrate‐binding site (M288, M397, F448) to open space for substrate binding and facilitate synthesis of the unnatural amino acid. A series of mutant plasmids were generated using site‐directed mutagenesis and sequenced to confirm the mutation. The wild type and mutant enzymes were recombinantly expressed in E. coli, purified by affinity and ion‐exchange chromatography, and purity was analyzed by gel electrophoresis. Enzyme activity with tyrosine in an NADH‐dependent coupled enzyme was first used to determine that the wild type and mutants are active, and initial results for the first mutants generated (M228S and M379A) indicate that the mutants are active. We are currently surveying reaction conditions using high‐performance liquid chromatography (HPLC) to evaluate the reactions of the TPL mutants with naphthols as substrates. HPLC is being used to determine the generation of product to calculate yields and provides a method to test if additional products are formed. While a goal of this study is aimed at the more efficient production of tyrosine derivatives, a larger goal is to better understand the features important for substrate binding in enzymatic reactions. This understanding may aid in the development of enzymes with a broad range of substrates efficiently converted to products in the biochemical generation of complex molecules.Support or Funding InformationFunding by NIH/NIGMS T34GM008074
Not all cartilages are the same! Differential effects in the cranial vs caudal skeleton after BMP inhibition in zebrafish
Authors: Franz-Odendaal Tamara Anne , Zinck Nicholas 
Published: April 2020

Several signaling factors are known to be involved in cartilage morphogenesis via the regulation of condensation size, shape, and location. Of these signaling factors, the bone morphogenetic proteins (BMPs) are known to be crucial in regulating the formation of condensations, cellular differentiation, and in the expression of the downstream transcription factors. Furthermore, interference with the BMP signaling pathway results in disrupted bone and cartilage development in various vertebrates. This study aims to elucidate the effects of BMP inhibition on cartilage morphology in the zebrafish. We utilise a pharmaceutical BMP pathway inhibitor at specific early time points of development. We focus on two cartilages, one cranial and one caudal, that develop concurrently but which have different developmental origins. Our data shows that the cranial cartilage, namely the scleral cartilage, is highly robust and unaffected by the BMP inhibitor. This is in stark contrast to the caudal cartilages, for example the hypurals, which are dramatically affected. These effects include fusion of elements, missing elements and altered morphology. Understanding these variable effects provides insight into the mechanisms that limit versus enable cartilages from altering their shape and/or size. The robustness of the cranial skeleton is particularly noteworthy given the multitude of disorders that affect this part of the skeleton.Support or Funding InformationFunding was provided by the Natural Science and Engineering Research Council of Canada
Etiology of disrupted cholesterol metabolism in the liver of young rats submitted to obesogenic diets
Authors: St-Pierre David H. , Saint-Amand Roxane , Ngo Sock Emilienne T. , Lavoie Jean-Marc 
Published: April 2020

The purpose of the study was to test the hypothesis that a short period (2 weeks) of a high fat (HFD) and a western (WD) diet leading to an increase in liver fat accumulation will disrupt key markers of liver cholesterol metabolism mainly targeting the LDL‐receptor (LDL‐R) pathway. Rats were submitted to either a standard (SD), HFD (60%) or a WD (40% fat + 35% sucrose) diet for 2 and 6 weeks. Body weight gain and body fat values were higher (p < 0.01) in rats fed the HFD and WD for 6 weeks although food intakes in kcal/d were not different for all groups throughout the experimental period. Liver fat accumulation was significantly (P < 0.01) higher in HFD and WD rats as compared to the SD diet for both feeding durations. On the other hand, liver fat accumulation was higher in WD rats (P < 0.05) than HFD animals after two weeks of treatment. In addition, WD animals displayed the highest plasma cholesterol levels. After 2 and 6 weeks of treatment, hepatic gene expression of LDL‐R, Pcsk9 and SREBP‐2 was lower (p < 0.01) in WD animals than in HFD and SD rats. In addition, mRNA levels of Lrp1 and Acat2 were lower (p < 0.01) in WD rats as compared to those fed with SD. Gene expression of HMGCoA‐r, the main enzyme involved in the regulation of cholesterol synthesis, was lower in HFD (2 weeks) and even more so in rats fed the WD diet (2 and 6 weeks). The WD was associated with a higher (p < 0.01) gene expression of markers involved in the mediation of de novo lipogenesis (ACC and ChREBP). It is concluded that a short‐term WD promptly increased fat accumulation while disrupting cholesterol metabolism in the liver. The present results provide new insights on the mechanisms through which westernized diets rapidly promote increases in plasma cholesterol levels.Support or Funding InformationNSERC, FRQS, CFI
Review of Riboflavin Transporter Deficiency Syndrome: Sameness in Genotype occurs with Variations in Phenotype.
Authors: Ukpong Christopher , Einstein George P , Tulp Orien Lee 
Published: April 2020

Riboflavin Transporter Deficiency Syndrome (RTD) is a rare childhood neurodegenerative disorder. RTD is acquired as an autosomal recessive pattern, which indicates that a child whose parents are carriers of RTD has a 25% chance of having the disease [homozygous for the RTD trait], a 50% chance of being a carrier [a heterozygous for the RTD trait], and a 25% chance of not having the disease [homozygous negative for the RTD trait]. RTD is a sensory motor neuron disease with symptoms such as auditory neuropathy, sensory ataxia, sensorineural deafness, and muscle weakness. These symptoms were first described in 1892 but diagnosis only became established in 2009 following the application of molecular biology investigation to discern the cause. Through exome sequencing and genetic testing, a mutation of the Solute Carrier 52A gene (SLC52A) was found to be implicated in RTD. There is no cure for RTD currently, but dietary Riboflavin supplementation has been shown to be a very effective treatment for most RTD patients. The current treatment is based on mg Riboflavin/kg of Body Weight/day. This approach fails to consider the role of other potential factors in the efficacy of the therapy, as well as in the variations and severity of the symptoms. This research uses case reports and meta‐analysis of available work reports to highlight the potential pathophysiological factors that may contribute to the variations, severity of signs and symptoms reported, and treatment responses in RTD affected patients. This report includes the molecular, cellular, and the phenotypic dynamics of RTD, and ends with a review of reported treatment responses. It is believed that such a hierarchical assessment will better equip the patients, their families, and the attending clinicians with the knowledge to assess and treat each RTD patient individually and with greater accuracy, success and outcomes than in the past.Support or Funding InformationThis research supported by Institutional Resources of the University of Science Arts and Technology, Montserrat, British West Indies and the Einstein Institute, USA.
Ring‐14 Chromosomal Syndrome: A Statistical Analysis and Comparison of Signs and Symptoms
Authors: Ivanoff Athena E. , Tulp Orien Lee , Einstein George P. 
Published: April 2020

The parent(s) and/or guardian(s) of Ring‐14 patients all signed appropriate consent form(s) prior to the study.OBJECTIVEThe purpose of this extensive study was to investigate the manifestations and challenges faced by subjects suffering from the rare, genetic condition known as Ring‐14 Chromosomal Syndrome, caused by an abnormal ring formation of chromosome fourteen (14), by comparing signs and symptoms exhibited, including, but not limited to the following: phenotypic similarities; physical appearances and properties; abnormal oral manifestations (such as delayed development and eruption of teeth, excessive swelling of the oral mucosa (periodontitis and gingivitis), and presence of any atypical oral lesions and /or sores), maxillofacial characteristics; specific types of seizures most commonly experienced; onset, duration and frequency of seizures; most common race(s) affected; comprehension capabilities (such as the ability to speak, read, or write, and IQ/intelligence quotient), gender; age, presence of lymphedema; mobility characteristics; and existence and range of decreased muscle tone. The parent(s) and/or guardian(s) of Ring‐14 patients all signed appropriate consent form(s) prior to the study, which was also approved by the Human Subjects Committee of the University. The information gathered and analyzed from this study is intended to raise an awareness of this extremely rare, yet highly complex genetic condition and allow health professionals to gain familiarity with the signs and symptoms exhibited by these patients to encourage genetic testing early on, upon suspicion of it's potential presence; thus, limiting the occurrence of a late, or misdiagnosis. In addition to educating health professionals and those in training to appropriately examine these patients, as many of them are prone to seizures, behavioral outbursts, and are unable to speak; and finally, to challenge the current belief that this syndrome is limited to pediatric patients, by documenting and introducing current statistics of those affected by Ring‐14 Syndrome, as several cases remain diagnosed, yet undocumented in the current medical literature.Support or Funding InformationSupport provided by the University of Science Arts and Technology, Montserrat, British West Indies.
ECM1: A CTGF Binding Protein Inhibiting Ductular Reaction and Liver Fibrosis
Authors: Pi Liya , Ran Gai 
Published: April 2020

IntroductionExtracellular matrix protein 1 (ECM1) has inhibitory roles against transforming growth factor (TGF)β activation and liver fibrosis. We have identified ECM1 as binding protein for connective tissue growth factor (CTGF) in yeast two‐hybrid analysis. This study aims to determine whether ECM1 binding inhibits CTGF‐mediated ductular reaction and liver fibrosis.MethodsMice were fed with 3,5‐diethoxycarbonyl‐1,4‐dihydrocollidine (DDC) to induce ductular reaction and liver fibrosis. ECM1 was overexpressed in adenovirus and intravenously injected into animals before DDC feeding. ECM1 interaction with CTGF, and their effects on liver injury after DDC feeding were analyzed.ResultsECM1 downregulation was correlated with CTGF upregulation during DDC‐induced liver injury. ECM1 overexpression after DDC feeding caused low levels of EpCAM and Sox9 mRNAs, small areas of EpCAM+ cells, and few proliferating ductular epithelial cells in comparison to controls that received GFP adenovirus. Weak fibrogenesis was also observed in the Ecm1 overexpressing livers, as evidenced by low levels of fibrosis‐related genes, small fibrotic areas in staining with α smooth muscle actin antibody and Sirius red as well as low collagen production determined by hepatic hydroxyproline content. Finally, we found that ECM1 could interact with CTGF and integrin αvβ6 leading to low TGFβ activation in vitro and in vivo.ConclusionThese observations indicate that ECM1 is a novel binding protein for CTGF. Its overexpression significantly reduces DDC‐induced ductular reaction and fibrogenic reaction, supporting anti‐fibrotic potentials of this molecule for the treatment of chronic liver disease.Support or Funding InformationThis study is supported by National Institutes of Health NIAAA KO1AA024174 grant and Children Miracle Research Foundation grant awarded to L Pi.
Antihyperglycemic Potential of Butterfly Pea Leaf (Clitoria ternatea) Extract in Alloxan‐Induced ICR Mice
Authors: Malbas Fedelino Fernandez , Mondoy Melissa Undan , Trigo Blyth Barroga , Geronimo Josefina Garcia 
Published: April 2020

The present study was conducted to evaluate the antihyperglycemic potential of the hydromethanolic extract of Butterfly pea leaf in alloxan‐induced ICR mice.Thirty female ICR mice were grouped into six: normal, diabetic, positive control groups and three experimental groups treated with varying dosages of Butterfly pea leaf extract (100 mg/kg. b.w., 250 mg./kg b.w., and 500 mg/lg b.w.). All groups, except the normal control received alloxan monohydrate (200mg/kg b.w.) to induce diabetes. ICR mice were gavaged with their respective treatment’s once daily for 14 days and blood samples were collected weekly for the determination of blood glucose levels. On the 14th day, ICR mice were sacrificed and pancreases were extracted for histopathological examination.The administration of Butterfly pea leaf extract on alloxan‐induced ICR mice elicited a blood glucose lowering activity throughout the course of the experiment. The 500 mg/kg dose of the extract exhibited the highest percent blood glucose reduction at the two time intervals (69.79% and 81.92%). The three varying dosages of the extract showed statistically no difference (P>0.05) when compared with the positive control group/ Histopathological examination of pancreas reported no significant difference (P>0.05) in the pancreatic damage between the Butterfly pea leaves treated groups and the diabetic control group, thus may suggest a different mechanism of action responsible for the antihyperglycemic activity of Butterfly pea leaves . The above results may propose that leaf extract is a potential candidate as an antihyperglycemic agent.Support or Funding InformationResearch Institute for Tropical Medicine Veterinary Research Department, Alabang Muntinlupa City, Philippines; De La Salle University Laboratory Animal Research Center, Dasmarinas Campus, Philippines and Department of Education, Quezon City, Philippines
High Throughput siRNA Screening for Sulfur Mustard‐Induced Corneal Epithelial Cell Injury
Authors: Gomez Amber , Koenig Jeffrey , Ruff Albert 
Published: April 2020

Chemical injuries to the eyes are true ocular emergencies due to the potential to inflict significant tissue damage that can lead to vision impairment or eventual loss of vision. Sulfur mustard (HD) is a chemical warfare agent that was originally used in World War I and has been used as recently as 2016 in the Syrian conflict. As with many potential chemical threats, no specific treatment for HD ocular exposure is currently available. We previously utilized a high throughput small inhibitory RNA (siRNA) screening model to identify potential therapeutic targets for two different toxicant‐induced ocular injuries: chloropicrin and hydrogen fluoride. Several targets for these injuries are now being evaluated in mouse models of injury. We proposed utilizing the same high throughput screening (HTS) method to identify potential therapeutic targets involved in the response to HD‐induced ocular injury. The model utilized SV40 large T antigen immortalized human corneal epithelial cells (SV40‐HCEC). Selected endpoints were cell viability and IL‐8 expression, as targets of interest should mediate cell health and/or the inflammatory response. Primary screen results were used to down‐select 300 targets from the original 3,027. Deconvolution of these 300 targets narrowed the target list down to 25, which are currently undergoing in vitro validation. Various pathways are implicated by the current target list, including ubiquitination, transcription regulation, signal transduction, and membrane transport. We anticipate that approximately ten validated targets will be transitioned to study in a mouse model to further determine potential therapeutic efficacy for HD‐induced ocular injury. Although work for HD is still in progress, we have compared the target lists that entered in vitro validation studies for all three toxicants evaluated thus far. Only one target was found in common: NR3C1, a glucocorticoid receptor. Additionally, there were two targets in common between chloropicrin and hydrogen fluoride: KMT5A and RELA. These findings show little similarity in the early molecular responses to these toxicants and support the hypothesis that specific therapeutics will be needed to treat different chemical‐related injuries.Support or Funding InformationDisclaimer: The views expressed in this abstract are those of the authors and do not reflect the official policy of the Department of Army, Department of Defense, or the U.S. Government.Funding Sources: This research was supported in part by an appointment to the Postgraduate Research Participation Program at the U.S. Army Medical Research Institute of Chemical Defense administered by the Oak Ridge Institute for Science and Education through an interagency agreement between the U.S. Department of Energy and U.S. Army Medical Research and Materiel Command. Support is also provided by a CounterACT inter‐agency agreement between the NIH/NINDS (Y1‐O6‐9613‐01) and USAMRICD (A120‐B.P2009‐2).
Effects of Personalized Precise Exercise Training with Holistic Integrative Protocol (PPETHIP) on Patients with Chronic Heart Failure: Summary Report of Controlled Clinical Trial
Authors: Sun Xingguo 
Published: April 2020

BACKGROUNDThe effects of Personalized Precise Exercise Training with Holistic Integrative Protocol (PPET‐HIP) in patients with chronic heart failure (CHF) were less reported previously, we hypothesized that PPET‐HIP would improve exercise capacity, facilitate to manage co‐morbidities and decrease the average re‐hospitalization rate(ARR) among CHF patients.METHODS30 CHF patients were recruited and randomly divided into Control group (n=15) and PPET‐HIP group (n=15).Before and after 12‐week intervention, general evaluations (blood pressure, glucose, and lipid profiles), cardiopulmonary exercise testing (CPET), echocardiogram, 6‐min walking distance (MWD), quality of life (QoL) score, and the adjustment of drugs treated co‐morbidities of CHF patients in 2 groups were recorded respectively. The ARR of CHF patients in 3 years before and after intervention was analyzed respectively.RESULTSAfter 12‐week PPET‐HIP, the parameters including CPET parameters, left ventricular ejection fraction, 6MWD and QoL in PPET‐HIP group were improved significantly than those in control group or those before PPET‐HIP (P<0.05). The levels of blood pressure, glucose and lipid in PPET‐HIP group maintained stable after the dosages of drugs treated co‐morbidities were adjusted. Surprisingly, a few patients stopped taking their some drugs treated co‐morbidities. The ARR of cardiovascular‐related events in PPET‐HIP group was decreased obviously than that in control group (P<0.05).CONCLUSIONSBased on Holistic Integrative Physiology and Medicine, PPET‐HIP can improve exercise capacity and QoL, and can facilitate to manage the co‐morbidities of CHF, and to reduce ARR.Support or Funding InformationNational Natural Science Foundation of China (81470204); Fuwai Hospital, National Cardiovascular Institute of Chinese Academy of Medical Sciences (2012‐YJR02); Research on Clinical Characteristics of the Capital (Z141107002514084); Research and Outcome Promotion of Clinical Characteristics in the Capital (Project No. Z161100000516127)
PECAM‐1 and VEGF‐Dependent Permeability
Authors: Abraham Valsamma , Zoga Kristi , Desai Mitan , Massa Alyssa , DeLisser Horace 
Published: April 2020

VEGF has long been recognized as a potent permeability factor and PECAM‐1 has been implicated in endothelial barrier function. However, the role of PECAM‐1 in VEGF‐mediated permeability is unclear. We observed in studies of wild type and PECAM‐1‐null mice that the loss of PECAM‐1 was not associated with an increase in the baseline permeability of non‐stimulated vessels. Consistent with this was the finding that the junctional architecture of quiescent vessels was preserved in the PECAM‐1‐deficient mice. In contrast, VEGF‐dependent permeability was significantly impaired in the skin of PECAM‐1‐null mice compared to wild type animals, while the dermal responses to two other factors (PAF and histamine) were preserved in the mutant mice. In line with these in vivo findings, in vitro permeability induced by VEGF (but not that of PAF) was reduced in PECAM‐1‐null endothelial cells (ECs). Further, VEGF receptor phosphorylation and downstream activation of PI3 kinase, Akt and eNOS following VEGF stimulation were blunted in PECAM‐1‐null ECs. Together these data implicate the involvement of PECAM‐1 in VEGF‐mediated permeability.Support or Funding InformationCompetitive Pilot Project Fund, VA Health Network VISN4