bluuebunny/crossref_metadata_2025_split huggingface dataset:
TrmB an archael transcriptional regulator
Authors: Thomm Michael , Surma Melanie 
Published: June 2021

In Pyrococcus furiosus two binding protein–dependent, substrate–specific ABC transporters could be identified, one of it being a trehalose/maltose (TM)–transporter, which is responsible for the uptake of trehalose and maltose. By binding to the respective operator sites TrmB prevents the further binding of transcription factors and subsequent RNA polymerase recruitment, which leads to an inhibition of transcription. With the aid of in vitro transcription assays using purified components of P. furiosus it could be shown that TrmB–mediated inhibition of transcription on both promoters is released by specific carbohydrates with maltose and trehalose being inducers for the TM operon, and maltodextrins as well as sucrose releasing inhibition on the MD promoter.TrmBL1 is homologue to TrmB and is involved in the regulation of genes encoding glycolytic and gluconeogenetic enzymes, respectively, where it recognizes a conserved sequence motif called TGM. It inhibits transcription of glycolytic genes with maltose, maltotriose and fructose as inducers (3). On the other hand TrmBL1 activates the expression of gluconeogenetic genes as well. Since the TGM is not present in the promoter region of the TrmBL1 gene, the TGM is not essential for TrmBL1 target recognition (3).Both TrmB and TrmBL1 share the unique property of recognizing different DNA sequences on different promoters and showing distinctive, promoter–dependent sugar specificity.In vitro transcription assays, EMSA and DNaseI footprinting experiments proved that TrmB and TrmBL1 have at least two different HTH motifs and recognize their respective target promoters with different helices within their N–terminal DNA binding domains.
Overexpression of recombinant human procathepsin B in E. coli
Authors: Wen Lisa , Chmielowski Jennifer , McConnell Matthew , Hunter Jessica , Keefer Ryan , McConnell Rose 
Published: June 2021

Cathepsin B is a lysosomal cysteine proteinase that is implicated in the processes of tumor invasion, metastasis, and Alzheimer's disease. Development of specific inhibitors of cathepsin B could lead to the discovery of therapeutic agents for treatment of many types of carcinomas, as well as Alzheimer's disease. The objective of this research is to produce functional recombinant cathepsin B enzyme which will be used in screening of new cathepsin B inhibitors. Human procathepsin B cDNA has been successfully amplified by PCR and sub‐cloned into pET15b expression vector. Overexpression of the procathepsin B protein has been accomplished by IPTG induction. The ability to produce large quantities of procathepsin B enzyme will permit a drug discovery program for development of newer effective inhibitors of cathepsin B.Supported by National Cancer Institute at NIH (Grant No. 3R15CA086933‐04 and 3R15CA086933‐04A2S1)
Order in Disorder: Aggrecan CS Region Predicts a New Class of Protein Structure
Authors: Hardingham Timothy Edward , Jowitt Thomas , Murdoch Alan , Day Joanna , Berry Richard , Baldock Clair 
Published: June 2021

Aggrecan is a glycosylated multi‐domain protein. Secondary structure analysis of the extended CS sequence predicts no known folded motifs. It is assumed to be unstructured and its elongation caused by high glycosylation. We expressed a 31KDa CS‐peptide sequence from human aggrecan in E Coli and compared its unglycosylated structure with human aggrecan globular G3 domain.The CS‐peptide and globular G3 domain were analyzed by MALLS/SEC, SAXS, sedimentation velocity and CD (Table 1). The CS‐peptide was hydrodynamically much larger than G3 and its sedimentation coefficient 1.67S was much less than predicted (2.1S) for a random‐coil. This suggested that the CS‐peptide was very extended, and its sedimentation rate increased and CD spectrum changed in 6 M urea suggesting a significant collapse of secondary structure. Modelled as a segmented flexible chain, (Kratky/Porod and Holtzer) gave a segment length 3.3 nm, with 13 repeats and the calculated mass per unit length of 831 gmol‐1 nm‐1 was also much higher than an extended polypeptide chain (308 gmol‐1 nm‐1).

Mass (Da)
Mass (Da) exptl
S20, W
f/f0
Rh (nm)
Rg (nm)

G3
25700
26800 ± 700
2.38 ± 0.14
1.31
2.57 ± 0.17
N/D

CS‐peptide
30827
28670 ± 1787
1.67 ± 0.11
2.01
3.99 ± 0.28
4.71

The CS‐peptide of aggrecan thus has an elongated and stiffened structure even when unglycosylated and it identifies a new class of semi‐ordered structure within disordered sequences. As 50% of all expressed protein sequences in the genome are predicted to be disordered these semi‐ordered sequences may be more commonly found than is currently recognized.
Characterization of fluorescent probe partitioning in giant unilamellar vesicles of “lipid raft” mixtures using confocal fluorescence microscopy
Authors: Sharom Frances J. , Juhasz Janos , Davis James H. 
Published: June 2021

Direct visualization of raft‐like liquid ordered (lo) domains requires fluorescence probes with known partitioning preference for a specific lipid phase. Therefore, a detailed understanding of the behaviour of commonly used fluorescent probes in defined lipid bilayer systems is needed before they can be used to draw conclusions about the physical state of the membrane. Using giant unilamellar vesicles composed of a ternary “lipid raft” mixture (DOPC/DPPC/cholesterol) for which the phase behaviour is known, we examined nine commonly used fluorescent probes using confocal fluorescence microscopy. The partitioning preference of each probe was assigned using either a well‐characterized ld phase marker, or by quantitation of the domain area fraction. Fluorescent molecules were examined individually, in pairs, and in threes; most of them partitioned into the ld phase, while two probes preferred the lo phase or gel phase. Interestingly, the partitioning of DiIC18 was influenced by Bodipy‐PC, and Lissamine rhodamine B‐DPPE affected the partitioning preference of other fluorescence probes when used with them. We compare and contrast the different fluorescence probes in terms of their partitioning preference, ability to detect phase separations, photostability, and any induced change in lipid miscibility transition temperature.Supported by the Natural Sciences and Engineering Research Council of Canada
The quantitative monitoring PtdIns(4,5)P2 dynamics in living cells using a specific molecular sensor
Authors: Yoon Youngdae , Lee Park Joo , Cho Wonhwa 
Published: June 2021

Phosphatidylinositol‐4,5‐bisphosphate (PtdIns(4,5)P2), which is a minor component of the plasma membrane, plays important roles in many cellular processes, including signal transduction, exocytosis, endocytosis, and ion channel activation. A method for quantitatively monitoring the PtdIns(4,5)P2 concentration and dynamics in living cells is lacking at present. We have developed a new method of quantifying the PtdIns(4,5)P2 concentration in living cells using a specific PtdIns(4,5)P2 sensor made of the engineered ENTH domain of epsin1 labeled with an environment‐sensitive fluorophore. The PtdIns(4,5)P2 sensor allows robust quantitative determination of spatiotemporal fluctuation of PtdIns(4,5)P2 in giant unilamellar vesicles and mammalian cells by ratiometric analysis. The results demonstrate the spatially heterogeneous distribution of PtdIns(4,5)P2 in the plasma membrane and provide new insight into the fluctuation and dynamics of cellular PtdIns(4,5)P2 in response to various stimuli. Furthermore, this strategy is generally applicable to in situ quantification of other membrane lipids.Supported by NIH GM 68849 and GM76581
Ca ‐ signaling activation by complex nucleotide substance (PriMed) in mouse myoblast cells c2c12
Authors: Orlova Elena V. , Orlova Valentina S. , Baeva Julia I. , Schemyatovsky Alexandr N. 
Published: June 2021

In present paper PriMed was investigated as a potential therapeutic agent in stem cells therapy and particular metabolic intervention.Methodsc2c12 was cultured by typical experimental protocol. NO‐synthase in c2c12 was determined by EPR‐spectroscopy. For the fluorescence in c2c12 0.005% Pluronic was used.ResultsIt was shown that:
In c2c12 PriMed (0.005μg/μl) activated Ca2+‐dependent NOS in 47‐fold vs base level and in 4‐fold vs dinitrosyl iron complexes (DNIC) (1.8μM) and in more than 6‐fold vs ionomycin (20μM).
PriMed (0.005μg/μl) gave 1.5‐fold Ca2+ increase in cytoplasm vs DNIC(1.8μM) and 3‐fold vs base level in c2c12.
Thus, the below experiments suggest that there is a direct influence of PriMed on biochemical and metabolic pathways of myoblast cells.
Association of Angiotensin Converting Enzyme Insertion/Deletion and Angiotensinogen T235 Polymorphisms with Risk of Essential Hypertension
Authors: Hemimi Neveen Salah , Oda Naglaa Raafat , Hemedi Samira Ali 
Published: June 2021

Backgroundpolymorphism of renin angiotensin system has been proved to be implicated in the pathogenesis of several multigenetic diseases including hypertension.Aim of the workto investigate the linkage of I/D polymorphism of angiotensin converting enzyme (ACE) and Met to Thr variant at position 235 of angiotensinogen (AGT) to the incidence of essential hypertension.MethodSamples of blood were collected from 203 subjects with matched age and sex (110 hypertensive and 93 normal controls). The polymorphism of ACE and AGT were investigated by polymerase chain reaction techniques.ResultsAGT polymorphism showed significantly increased incidence of hypertension in TT genotype as compared to (MT& MM) genotypes (OR= 2.2, 95% CI= 1.0–5.1, P<0.05). Similarly the study of ACE polymorphism declared a minor risk for hypertension in DD compared to (DI& II) genotypes (OR= 1.2, 95% CI= 0.7–2.2). No significant association was found between DD genotype and other risk factors of hypertension, while there was association of TT genotype and increased serum creatinine and fasting blood sugar.ConclusionTT genotype of angiotensinogen polymorphism is a possible risk factors for hypertension.Source of Fund: Dubai Medical College
Alcohol Affects the Late Differentiation of Progenitor B cell
Authors: Ewing Jacquelyn Bowers , Chan Wendy , Wolcott Michael , Wang Hao 
Published: June 2021

BackgroundStudies showed in vitro alcohol exposure can affect the differentiation of progenitor B cell. This study investigates developmental stages on B cell differentiation and how alcohol affects these stages.MethodsOligoclonal‐neonatal‐progenitor (ONP) cells were cultured in vitro with/without the exposure of alcohol. Phenotype analysis was performed at various time‐points and the expression levels of transcription factors (TF) were also measured.ResultsONP cell differentiated into B220‐CD11b− and B220‐CD11b+ cells in 3 days. No statistically significant difference of B cell formation was found in both normal and alcohol groups when exposed to alcohol during the first 3 days. If alcohol was added at day 4 and exposed to the end of culture, few yielded B cells. The expression level of TFs was down regulated with 100mM alcohol.ConclusionsONP cells can differentiate into lymphoid or myeloid lineage but diverged at B220‐CD11b−/B220‐CD11b+ branch point. Alcohol affects the ONP cell differentiation into B lineage at a late stage after cell divergence into B220‐CD11b−. Alcohol down regulates TFs expression resulting in poor B cell differentiation.
Integrin α11 mediates CCN1‐induced focal adhesion and β‐catenin translocation in esophageal epithelial cells
Authors: Modak Cristina , Mouazzen Wasim , Nguyen Anthony , Pham Jennifer , Chai Jianyuan 
Published: June 2021

BackgroundCCN1 is a matricellular protein that plays a critical role in several normal and pathological processes by signaling through different integrin receptors in a cell‐type specific manner. Clinical studies have suggested CCN1 as a putative marker for malignant progression towards esophageal adenocarcinoma. However, little is known about its signaling mechanisms in this tissue. The aim of this study was to determine the integrin receptor that mediates CCN1 actions in esophageal epithelial cells.MethodsEsophageal epithelial cells were treated with recombinant CCN1 protein. Its effects on integrins were examined by microarray, real‐time PCR, immunoprecipitation, immunoblotting, and immunofluorescence microscopy.ResultsOut of all the known α‐ and β‐integrins, ITGA11 and ITGB5 were the only ones that responded to CCN1 treatment. Co‐immunoprecipitation assays demonstrated an association between integrin α11 and β5 in the presence of CCN1. Blocking either integrin subunit prevented CCN1‐induced focal adhesion and β‐catenin nuclear translocation. Taken together, these results provide evidence that integrin α11 can form a new partnership with β5 and that together they serve as a new receptor to mediate CCN1 actions in esophageal epithelial cells.This study was supported by the Department of Veterans Affairs of the United States and the American Heart Association.
PACAP induces axonal elongation in cultured monkey trigeminal ganglion cells
Authors: Nakajima Emi , Walkup Ryan D. , Shearer Thomas R. , Azuma Mitsuyoshi 
Published: June 2021

LASIK refractive surgery is widely accepted for correcting myopia. However, flap creation during LASIK surgery leads to complications including dry eye, caused by degeneration of sensory nerves. Our previous studies in rabbits showed that PACAP accelerated the extension of neuronal processes and recovered corneal sensitivity. The purposes of the present study were to 1) establish a cell culture system for monkey trigeminal nerve (TgN), whose fibers innervate cornea, and 2) determine if PACAP induces sprouting and elongation of axons. Expression of mRNA and cellular distribution for the PACAP receptor, PAC1, were determined by qPCR and immunostaining. Axonal length in cultured TgN ganglion cells was evaluated by staining with antibody for neurofilament. Our results showed that dissociated, cultured TgN ganglion cells contained neuronal and Schwann cells. The bioactive N‐terminal peptide PACAP27 significantly induced axonal elongation which was inhibited by the PAC1 antagonist PACAP 6–27. Downstream inhibitors of the PAC1 signaling pathway, AC and PKC, also inhibited axonal elongation. Together with positive PAC1 staining in neuronal cells, our data suggested involvement of PAC1 activation in axonal elongation. PACAP might be a candidate drug for treatment of complications after LASIK.Dr. Shearer receives consulting fees from, and Drs. Azuma and Nakajima are employees of, Senju Pharmaceutical Co. Ltd.
Effect of helicobacter pylori infection on iron absorption in asymptomatic adults consuming iron and zinc fortified wheat
Authors: de Romaña Daniel Lopez , Diazgranados Doricela , Pizarro Fernando , Ruz Manuel , Brunser Oscar 
Published: June 2021

IntroductionMicronutrient deficiencies and Helicobacter pylori (HP) infection are prevalent and co‐exist in several countries.ObjectiveThe objective of the study was to determine the effect of HP on iron absorption in asymptomatic adults consuming iron and zinc fortified wheat.Materials/MethodsHP was assessed using the C13‐urea breath test. On day 1 subjects received for breakfast bread fortified, per Kg flour, with either 55 mg ferrous sulfate and 60 mg zinc sulfate, labeled with 59Fe, as sulfate, or same amounts of ferrous fumarate and zinc oxide, labeled with 55Fe, as fumarate. On day 3 they received the other type of bread. On day 17 blood (20 ml) was drawn to assess circulating radioactivity. Iron absorption was determined using Eakins and Brown's double isotopic technique.Results/FindingsPrevalence of HP was 78%. Fourteen HP negative and 16 HP positive individuals completed absorption studies. There were no significant differences in the mean iron absorption of either ferrous sulfate or ferrous fumarate by infection status.ConclusionHP has no effect on iron absorption in asymptomatic adults consuming iron and zinc fortified wheat.Support by FONDECYT grant 1080032
Hemoglobin regeneration efficiency method revisited: an alternative bioassay for iron bioavailability
Authors: Yokoi Katsuhiko , Konomi Aki , Otagi Miki 
Published: June 2021

Iron (Fe) deficiency is a worldwide problem, and a search for foods containing bioavailable Fe is an effective measure. The slope‐ratio method using graded levels of dietary Fe vs hemoglobin (Hb) are commonly used for the estimation of the bioavailability of Fe in foods, but it requires a considerable number of rats and do not give a value for individuals. The objective was a comparison between the Hb regeneration efficiency (HRE) method and the slope‐ratio method.HRE methodTwenty‐one F344/N male weanling rats were fed a Fe‐deprived diet for 4 weeks to deplete body Fe stores. Then, three groups of seven rats each were repleted for 20 days using diets fortified with ferrous sulfate, ferric citrate, or cocoa powder (van Houten brand) each provided 20 mg Fe/kg diet. The diets were based on the AIN‐93G formula. HREs were 0.733, 0.350 and 0.336 for ferrous sulfate, ferric citrate and cocoa powder, respectively. The HRE of Fe in cocoa powder was 96% that of ferric citrate.Slope‐ratio methodAnother 32 weanling rats were depleted in Fe stores for 4 weeks and then 8 groups of four rats each were repleted for 25 days using graded levels of dietary ferric citrate or cocoa powder. The slope for cocoa powder was 99% that of ferric citrate.In summary, the HRE method gave a similar outcome to the slope‐ratio method. The bioavailability of Fe in cocoa was significantly less than that of ferrous sulfate and was similar to that of ferric citrate.
Association of cognitive impairment and zinc intake in HIV+ adults
Authors: Mandal Priyanka , Baum Marianna K , Sales Sabrina , Li Yinghui , Jayaweera Dushyantha T , Page John B , Campa Adriana 
Published: June 2021

BackgroundThe central nervous system contains 10% of all body zinc. Zinc deficiency, which is prevalent in HIV‐infection, has been associated with cognitive impairment. This study examined the risk factors associated with impaired cognitive performance in HIV infection.MethodsAfter consenting 231 HIV‐infected adults, a 30 point‐questionnaire assessed cognitive function by testing immediate memory, problem solving ability and quality of life. Those with <26 points were considered cognitively impaired. Blood was drawn for plasma levels of zinc and HIV disease parameters. Quality of life, alcohol and drug use, and 24‐hour dietary recall questionnaires were administered.ResultsThere were 54 cases (23.3%) with cognitive impairment. When compared to those without impairment, cases had significantly poorer dietary zinc intake (8.1±SD5.7 vs. 10.4±SD7.8 mg, p=0.027) and were significantly older (44.8±SD6.3 vs. 42±SD7.1 years, p=0.0088). Alcohol and illicit drug use and frequency of use, plasma levels of zinc, CD4 count<200 cell/mm3, gender, and quality of life scores were not significantly different between groups.ConclusionLower zinc intake is a risk factor for impaired cognitive performance in this cohort of HIV+ adults. Prospective studies are needed to examine whether zinc supplementation might alleviate cognitive impairment in HIV+ adults with zinc deficiency.Funded by NIDA
Lycopene suppressed leptin‐mediated malignant progression in human colon cancer cells
Authors: Lin Meng‐Chiu , Tang Feng‐Yao 
Published: June 2021

Colorectal cancer is one of the leading causes of cancer death in western countries. Previous studies indicate that leptin induces cellular proliferation through multiple signalling pathways in colon cancer cells. Recent studies suggested that loss of E‐cadherin adherent molecule has been associated with epithelial–mesenchymal transition (EMT) and the malignant progression of human colon cancer cells. Our previous studies indicated that lycopene could exhibit anti‐cancer effects on human colon cancer cells. However, the inhibitory effect of lycopene on leptin‐mediated malignant progression of human colon cancer has not been demonstrated well yet. The present study demonstrated that leptin dose‐dependently (0, 0.0125, 0.0625, 0.25μM) induced cellular migration and invasion through up‐regulation of matrix metalloproteinase (MMP)‐7 as well as suppression of E‐cadherin in human colon cancer HT‐29 cells. The molecular mechanisms of actions were through the modulation of MAPK/JNK and JAK/STAT‐3 signaling pathways. Furthermore, our results demonstrated that lycopene (2 μM) effectively inhibited leptin‐mediated cellular migration through up‐regulation of E‐cadherin molecule and modulation of JNK signaling pathway in human colon cancer HT‐29 cells. Take together; our results suggest that lycopene could act as an effective chemo‐preventive agent against the leptin‐ mediated malignant progression in human colon cancer cells.Grant Funding Source: National Science Council grant under agreement No. 97‐2320‐B‐039‐043‐MY3
Dietary inhibitors of monoamine oxidase A: quercetin could be a naturally‐occurring antidepressant
Authors: Ramsay Rona Ruth , Dixon Clarke Sarah E. 
Published: June 2021

Health supplements are frequently sought to alleviate low mood. The pharmacokinetics of components of tobacco and on flavonoids were examined to see whether a normal diet provides compounds active as monoamine oxidase inhibitors in sufficient quantities to affect mood. A literature search identified dietary monoamine oxidase inhibitors, their levels in foods, the pharmacokinetics of absorption, and the distribution and tissue levels observed. Estimated daily intake and expected tissue concentrations were compared with the efficacies of the compounds as inhibitors of monoamine oxidases from the literature and, for monoamine oxidase A, assayed spectrophotometrically.Norharman, harman and quercetin can be present in a healthy diet at up to 0.15 mg norharman, 0.11 mg harman and 54.2 mg quercetin per day. The pharmacokinetics and tissue levels for these compounds were consistent with significant levels reaching neuronal monoamine oxidase from the diet or smoking. Although diverse methods in the pharmacokinetic studies make the comparisons qualitative, quercetin was the highest in the sample diet and was competitively inhibited monoamine oxidase A. Quercetin in the diet could be a natural antidepressant compound.Supported by a Biotechnology and Biological Sciences Research Council Vacation Bursary.
Anti‐cancer effect of Korean radish(Raphanus sativus L.) leaf extract through induction of apoptosis and down‐regulate Erb B2 signaling and Akt pathway in human breast MDA‐MB‐231 cell lines
Authors: Chang Moon‐Jeong , Kim Boram , Lee Hye‐Sung , Kim Woo‐Kyoung , Cho Kangjin 
Published: June 2021

The vegetative portion of radish have not been characterized in terms of its biological activity. In vitro anti‐cancer effect of aqueous extract from Korean radish leaf (EKRL) was evaluated in human breast MDA‐MB‐231 cell lines. EKRL preferentially inhibited the growth of cell lines in a dose‐dependent manner (p<0.05). In addition, EKRL treated MDA‐MB‐231 decreased expression of anti‐apoptotic Bcl‐2 protein and unchanged expression of pro‐apoptotic Bax protein, eventually leading a decrease in the Bcl‐2/Bax ratio. Phosphorylated Akt levels markedly reduced in cells treated with 100ug/L EKRL and total Akt levels decreased at 300ug/L. Furthermore, a dose‐dependent decrease in Erb B2 and Erb B3 levels was detected in EKRL treated cells. These results demonstrated that EKRL inhibits the proliferation of MDA‐MB231 through induction of apoptosis and down‐regulate Erb B2 signaling and the Akt pathway, making EKRL a potent candidate as new‐anti cancer food components.(This research was supported by Korea Rural Development Adminstration, project #2009AFT143783066)