bluuebunny/crossref_metadata_2025_split huggingface dataset:
MRI and DTI characterization of spinal cord severe contusion injury in the rat
Authors: Lim Wootaek , Baligand Celine , Keener Jonathon , Ye Fan , Vohra Ravneet S , Ruhella Arjun , Bose Prodip K , Walter Glenn A , Thompson Floyd J , Vandenborne Krista H 
Published: June 2021

Spinal cord injury (SCI) induces loss of structural integrity of spinal cord tissue, resulting in locomotor deficit and muscle wasting. Thus, it is important to accurately assess the severity of injury. In this study, we characterized the spinal cord changes using diffusion tensor imaging (DTI) and 3D magnetic resonance imaging (MRI) following severe contusion injury. Sixteen‐week old female Sprague‐Dawley rats were given severe contusion injury at the T8 level using a NYU impactor (10g, 50 mm weight drop) (SCI, n=5). 3 animals were used for control (CTR). Ex vivo 3D MRI and 2D DTI were performed 3 weeks post‐injury in a Bruker 750MHz magnet. Imaging parameters were: field of view of 4.5 × 4.5 mm2; matrix size of 128 × 128; in‐plane resolution of 35.16 × 35.16 μm2. Images were acquired over 1 cm of the length of the cord, including the epicenter. All procedures were approved by the University of Florida institutional animal care and use committee. In CTR, spinal cord cross‐sectional was 4.38 ± 0.21 mm2 on average. This value was decreased to 1.58 ± 0.09 mm2 at the epicenter in SCI 3 weeks post‐injury. Fractional anisotropy (FA) for CTR was 0.76 ± 0.01 in white matter and 0.51 ± 0.05 in gray matter with a mean FA of 0.72 ± 0.02. In SCI, mean FA was decreased to 0.40 ± 0.09 at the epicenter. These findings demonstrate that the injured cords show lower FA value with significant atrophy following severe contusion injury.Grant Funding Source : NIH Grant P01 HD059751
Z‐Bisdehydrodoisynolic Acid (Z‐BDDA), a putative estrogenic seco‐steroid, induces profound lordosis behavior in ovariectomized female rats
Authors: Viscomi Natalie S , Clough Rich W , Arbogast Lydia , Banz William J , Hou Yuqing , Meyers Cal 
Published: June 2021

Z‐Bisdehydrodoisynolic Acid (Z‐BDDA) has been described as highly estrogenic, and a selective estrogen receptor modulator, based exclusively on uterotrophic activity in female rats. Exploring possible neuroprotective effects of Z‐BDDA, we thought it important to determine whether Z‐BDDA has estrogenic effects in the brain. Accordingly, female Sprague‐Dawley rats were ovariectomized (ovx) under isofluorane anesthesia and assigned to various steroid treatment protocols intended to induce sexual receptivity. Z‐BDDA (300ug/rat or 30ug/rat) or Estradiol benzoate (10 ug/rat) was administered (sc) two weeks following ovx and some rats were also given Progesterone (1mg/rat) 48 hours later. Lordosis behavior was assessed by placement of female rats in an arena containing a sexually mature male rat. Mount attempts by the male and lordotic responses in the females were recorded. Results showed that female rats treated with either Z‐BDDA or Estrogen (followed or not by Progesterone) exhibited profound receptivity assessed by display of lordosis in response to male mount attempts. Thus, this study further establishes the estrogenicity of Z‐BDDA and demonstrates a positive feedback effect on the central nervous system that appears to parallel the peripheral (uterotrophic) effects of the compound. Supported by the SIU School of Medicine Center for Integrated Research in Cognitive and Neural Sciences.Grant Funding Source : None
Role of the Ccr4‐Not Complex in Histone Acetylation
Authors: Mehrotra Swati , Vancura Ales 
Published: June 2021

The Ccr4‐Not complex is a global regulator of gene expression that affects expression both positively and negatively. The complex, conserved from yeast to human, is composed of at least nine subunits. The Ccr4 subunit functions in the control of mRNA deadenylation and transcriptional initiation/elongation. The NOT proteins are involved in transcriptional repression by restricting access of TFIID to promoters. The Ccr4‐Not complex also plays a key role in sensing nutrients and stress. Our earlier results implicated Ccr4‐Not complex in regulation of global untargeted histone acetylation. This notion is also supported by the synthetic negative genetic interactions between the ccr4Δ and not4Δ mutations and mutations in SAGA and NuA4, the main histone acetyltransferases in yeast. The histone hypoacetylation phenotype of ccr4Δ and not4Δ mutants can be partially suppressed by attenuated expression of acetyl‐CoA carboxylase ACC1. This result suggests that ccr4Δ and not4Δ mutants have altered regulation of intermediary metabolism, reduced level of nucleocytosolic acetyl‐CoA, and consequently display histone hypoacetylation. Together, our data highlight the connection between intermediary metabolism and histone acetylation and indicate that the nucleocytosolic level of acetyl‐CoA is an important determinant of histone acetylation.This work was funded by NIH.
Changes in genes methylation of the TLR pathway in livers of mice fed DDC
Authors: Oliva Joan , French Samuel W. 
Published: June 2021

BackgroundMallory‐Denk body (MDB) formation is a component of alcoholic and non alcoholic hepatitis. Proteins of the TLR pathway were shown to be involved in the formation of MDBs, in mice fed DDC. TLR genes are upregulated and SAMe supplementation prevents this up regulation and prevented the formation of MDBs.MethodDNA of livers from control mice, from mice fed DDC 10 weeks, refed 1 week with DDC and with DDC+SAMe were extracted and used to study the methylation pattern of genes involves in the TLR pathway. A PCR array was used to analyze it.ResultsUsing PCR array for the mouse TLR pathway (24 genes) were found whose the expression of IL12A was regulated by the methylation of its gene. DDC treatment reduced the methylation of the IL12A gene expression, with a similar level when DDC was refed. However, in the presence of SAMe, the intermediate level of methylation of IL12A was up regulated and the methylation of the promoter decreased compared to DDC refeeding or DDC 10 weeks.ConclusionIL12a is known to induce the production of IFNg by NK and LT. We showed in previous publication that IFNg is one of the major cytokines involved in the induction of MDB formation. The low expression of IL12A associated with the intermediate methylation of its promoter could explain one step in the mechanism which leads to the formation of MDBs. Supported by NIAAA PA50‐011999.
Towards predicting the behavior of E. coli LacI and FadR transcription factors to variations in their operator sequences
Authors: Maity Tuhin Subhra , Jha Ramesh , Strauss Charlie , Dunbar John , Kuske Cheryl 
Published: June 2021

Transcription factors (TFs) typically activate or repress gene expression by binding to specific DNA sequences in the genome called operators. To deeply understand this process, it is necessary to catalog the behavior of transcription factors to variations of their operator sites. This offers the potential for recognition of alternative binding sites, and, synthetically, offers a prescription for tuning binding affinity. Ultimately, one hopes to acquire a predictive understanding of the sequence‐behavior relationship and use that knowledge for metabolic engineering.In this study, we evaluated the effect of operator DNA sequence changes to the transcriptional repression properties of two E. coli TFs, LacI and FadR. We started with a known DNA operator sequence of the E. coli LacI TF and discovered the preferred FadR binding DNA sequence using a protein‐binding microarray experiment, and then randomly mutated a small subset of the nucleotides in their operator sites. We cataloged the effect of sequence change to the transcriptional repression/induction properties by the two TFs using an in vivo GFP reporter assay. For both the TFs, we observed a wide range of interesting and characteristically distinct repression and induction behaviors, and some of them exhibited a strong correlation with a specific nucleotide at a specific position of the operator DNAs. We also performed Rosetta modeling calculations to estimate relative TF‐binding energies of our operator variants from the existing 3D structures of the TF‐DNA complexes. Our results suggest that 3D structure calculations can infer the behavior of a useful subset of the sequence variations, but is not as yet sufficient to completely explain the behavior of all sequence variations.
Human Cosmc and T‐synthase Genes Are Transcriptionally Regulated by SP1/SP3 Transcription Factors
Authors: Ju Tongzhong , Mi Rongjuan , Wang Yingchun , Ding Xiaokun , Song Lina , van Die Irma , Chapman Arlene B. , Cummings Richard D. 
Published: June 2021

BackgroundCosmc is the specific molecular chaperone for core 1 β3‐galatosyltransferase (T‐synthase), which is the key enzyme for synthesizing the core 1 structure in mucin type O‐glycans. Interestingly, human Cosmc and T‐synthase are ubiquitously and coordinately expressed but vary by cell and tissue type. Genetic mutations in Cosmc have been identified in several human diseases. Recent studies have also shown that both Cosmc and T‐synthase are transcriptionally suppressed in the B cells from some patients with IgA nephropathy. Yet, how these two genes are regulated is unknown.Study ObjectiveIdentify and characterize the promoters for human Cosmc and T‐synthase.MethodsLuciferase reporter assay and chromatin immunoprecipitation (ChIP) analysis.Resultsthe proximal GC‐rich region (CpG island) at the 5′‐flank of human Cosmc (cCpG‐II) which contains two Specificity Protein 1 and 3 (SP1/3) binding sequences has the promoter activity. Similarly, the tCpG in human T‐synase which also contains two SP1/3 sites is the promoter for T‐synthase. Site direct mutagenesis demonstrated that both SP1/3 sequences for Cosmc and T‐synthase are essential to their promoter activity. Furthermore, ChIP analyses confirm that the putative SP1/3 sequence in the promoter associates with the SP1/3 for both Cosmc and T‐synthase.ConclusionHuman Cosmc and T‐synthase are transcriptionally active at a basal level by SP1/SP3.
Proposal for international nutrition criteria for school lunches for children aged 5–18
Authors: van Elk Kathelijn Neeltje , Eilander Ans , Roodenburg Annet JC 
Published: June 2021

The nutritional status of children aged 5–18 is often suboptimal and across the globe, school lunches have an important contribution to children's daily diet. Therefore, providing a nutritious school lunch can likely have a significant impact on children's health. However, for the development of nutritious meals, international science‐based nutrition criteria are currently lacking for both primary and secondary schools.AimTo develop international criteria for the nutritional composition of school lunches.MethodsFAO/WHO recommendations for total daily diets were translated into nutrient criteria for school lunches. Specific criteria were developed for total energy, proteins, total fat, PUFA, SAFA, trans fatty acids, carbohydrates, added sugars, and sodium, as well as for the critical micronutrients folate, zinc, iron, iodine, vitamin A, and vitamin D, which are often suboptimal in children's diets.ResultsCriteria were defined so that a lunch meal delivers 30% of the daily energy intake and 50% of the micronutrient recommendations for children aged 5–18. Tolerable upper intake levels were taken into account.ConclusionWhile criteria were developed globally, implementation will differ between countries based on differences in dietary habits, affordability and local food composition. Also, feasibility of developing recipes based on the criteria needs to be further explored.
Biomarker in human breast cancer cells
Authors: Schmidt Jane , Schmidt Jane 
Published: June 2021

Multiplex measurement of biomarkers in human breast cancer cell culture supernatesJane Schmidt, Ph.D, Michael Hall, Richard Fuerstenberg. R&D Systems, Inc. Minneapolis, Minn.The mixture of biomarkers that is produced in the tumor microenvironment has an important role in cancer pathogenesis. For example, cytokines that are released in response to infection, inflammation and immunity can function to either promote or inhibit tumor development and progression. The multiplex immunoassay has become an efficient tool both for rapid screening and more in‐depth analysis of cancer cell supernates, while requiring only a single, small sample volume. In this study, the VersaMAP™ Development System was used to rapidly and simultaneously screen for human biomarkers in supernates from six human breast cancer cell lines. Fifteen positive biomarkers were identified and biomarker profiles were established for each of the cell lines. The VersaMAP Development System, developed by R&D Systems,Inc., is a customized assay system designed for the simultaneous measurement of multiple biomarkers from a single 50 μL sample. It offers the ability to measure up to 10 biomarkers chosen by the user from a continuously expanding library of 39 available analytes. The assay is available in a custom premixed form, giving the user total flexibility to choose their multiplex panel. The assay system is a bead‐based multiplex immunoassay designed for use with a Luminex® or Bio‐Rad® Bio‐Plex® analyzer.
Influence of physical exercise on cardiac structure and function of spontaneously hypertensive rats
Authors: Pagan Luana Urbano , Damatto Ricardo Luis , Cezar Marcelo Diarcadia Mariano , Gimenes Rodrigo , de Campos Dijon Henrique Salomé , Lima Aline Regina Ruiz , Bonomo Camila , Guizoni Daniele Mendes , Okoshi Marina Politi , Okoshi Katashi 
Published: June 2021

The aim of this study was to evaluate the influence of physical exercise (PE) on cardiac structure and function of spontaneously hypertensive rats (SHR) before clinical evidence of heart failure (HF).Methods16 month old SHR were divided into two groups: sedentary (SED, n=14) and exercised (EX, n=14). Physical training was performed 5 times/week for 16 weeks in a treadmill. Arterial pressure (AP) was measured by tail‐cuff method. Echocardiogram (Echo) was performed to evaluate in vivo cardiac structures and left ventricular (LV) function. In vitro myocardial function was analyzed in LV papillary muscle preparations. Statistics: Studentˈs t test (p<0.05).ResultsAP did not differ between groups (SED: 212±22; EX: 225±20 mmHg). Echo did not show differences in cardiac structures or LV systolic function. Late diastolic mitral annular velocity (Tissue Doppler) was lower in EX group (3.3±0.7 vs 4.2±1.1 cm/s). The in vitro study showed increased contractile function in EX group (developed tension: 7.6±1.8 vs 6.2±1.1 g/mm2; +dT/dt: 67±18 vs 53±10 g/mm2/s).ConclusionPE started before clinical evidence of HF is beneficial on LV diastolic and myocardial contractile functions in SHR. Support: CAPES, CNPq.
The prostate in DMBA administration – Morphometric evaluation
Authors: Barandas Gustavo Teixeira , Pereira Filipe , Costa Eduardo , Farinha Rodrigo , Cabrita Silverio 
Published: June 2021

7,12‐Dimethylbenz(a)antracene (DMBA) is a xenobiotic found in cigarette smoke and exhaust gases of combustion engines. It is an aromatic polycyclic hydrocarbon, metabolized by P450 enzymes producing reactive metabolites which form covalent adducts with DNA and the formation within DNA of depurinated abasic sites. The aim of this study is the evaluation of DMBA action on prostate by morphometric and histopathological analysis.Ten Wistar male rats, 8 weeks old, were submitted to DMBA administration, 20 mg/Kg/day, three times a week. The control group was kept without manipulation. All the animals were killed after two weeks, and a complete necropsy was performed. The prostate fragments were stained by H&E and the pieces lined by epithelia were measured using the software Image J. The frequencies distribution of the control group were achieved and considered as a reference value, for test group comparison. No statistic significance was found. The histology evaluation of the prostate of test group show no morphologic lesions.These results allow us to suggest that the acute intoxication with DMBA, as in this model, has no morphologic translation.
Highly palatable food normalizes the HPA axis activity and modulates anxiety‐like behaviors in rats that experienced neonatal maternal separation
Authors: Jahng Jeong Won , Kim Jin Young , Park Eunyoung , Lee Joo Young , Lee Jong-Ho 
Published: June 2021

In this study, we have examined the effect of highly palatable food (HPF) on the psycho‐emotional behaviors of rats that experienced neonatal maternal separation. Male SD pups were separated from dam for 3 h daily during PND 2–14 (MS) or left undisturbed (NH). Half of NH and MS pups had free choices of cookie (HPF) and chow from PND 28, the rest half chow only, and were subjected to behavioral tests during young adulthood. Plasma corticosterone levels were analyzed by radioimmunoassay, and tissue contents of ΔFosB and pCREB in the nucleus accumbens by western blot. Caudal grooming was reduced in MS‐control and NH‐HPF compared with NH‐control. HPF suppressed defecation activity in MS, but not in NH. HPF increased anxiety‐like behaviors during elevated plus maze test and depression‐like behaviors during swim test in NH, and these behaviors were not further affected by HPF in MS. Stress‐induced corticosterone increase was blunted by MS experience, and HPF appeared to normalize it. pCREB in the nucleus accumbens was increased, and ΔFosB decreased, by MS experience, and HPF normalized them. Results demonstrate that long‐term HPF may normalize the HPA axis activity and modulate anxiety‐like behaviors in MS rats, likely in relation with neural plasticity in the nucleus accumbens.Supported by the 21st Century Frontier Research Program (2009K001269) and the NRF (2010‐0003642) funded by MOEST
Protease‐activated receptor 1 expressed in the human corpus cavernosum induces phosphorylation of endothelial nitric oxide synthase at threonine 495
Authors: Mantey Jane Ama , Motley-Johnson Evangeline 
Published: June 2021

Protease‐activated receptor 1 (PAR1) can induce phosphorylation of endothelial nitric oxide synthase (eNOS) at site threonine 495 to inhibit production of nitric oxide. The objective of this project is to elucidate whether PAR1 is a potential therapeutic target for improving availability of nitric oxide in patients with erectile dysfunction and related vascular diseases. PAR1 has not been shown to be expressed in the penile endothelium, however. I hypothesize that PAR1 is expressed in the endothelium of the corpora cavernosa of the penis and, once activated, leads to significant phosphorylation of eNOS‐Thr495. Human corpus cavernosum endothelial cells (HCCECs) were lysed and probed for PAR1 expression using western blot and immunocytochemistry. The cells were then stimulated with thrombin or TFLLR, a PAR1 agonist, and phosphorylation of eNOS was examined by western blot and enzyme‐linked immunosorbent assay. Expression of PAR1 was detected in the HCCECs. Treatment with thrombin and TFLLR induced concentration and time dependent phosphorylation of eNOS‐Thr495. This study shows that PAR1 is expressed in the penile endothelium. Future studies include analyzing any differences in expression or functionality of PAR1 in HCCECs from males with erectile dysfunction.This project was supported, in part, by National Institute of General Medical Sciences (NIGMS) Grant 5SC3GM086336‐02.
Novel mechanisms to ATP‐dependent glucose uptake in skeletal muscle cells
Authors: Osorio-Fuentealba Cesar , Contreras-Ferrat Ariel , Klip Amira , Jaimovich Enrique 
Published: June 2021

Contraction and insulin stimulate glucose influx into muscle, and contraction further sensitizes muscle to insulin. Insulin signals via α,β class I phosphatidylinositol 3‐kinase (PI3Kα,β) to Akt, but contraction signals remain elusive. Interestingly, ATP is released by contracting muscle, and it was recently reported that extracellular ATP can act auto/paracrinely on muscle cells. Our aim was to investigate if ATP induces glucose uptake in muscle cells and promotes insulin action.Rat primary myotubes transiently transfected with GLUT4myc and L6 myoblasts stably expressing GLUT4myc were exposed to 9s 6V electric stimulation (ES); insulin (100 nM, 5 min) or ATP (100 uM, 5min). Uptake of the fluorescent hexose 2NBDG was assayed in real time. Surface GLUT4 was measured by anti‐myc antibodies in fixed, unpermeabilizad cells. Phosphorylation of Akt and its target AS160 was detected with site‐specific antibodies.ES, insulin and ATP each stimulated glucose uptake, GLUT4 translocation, Akt and AS160 phosphorylation. ATP added prior to insulin increased glucose uptake beyond insulin's effect. Inhibitors of Akt (AktVIII), PI3Kα–γ (LY294002), PI3Kγ (AS605240) or dominant‐negative PI3Kγ and Akt inhibited the ATP‐effect. In sum, ES or ATP promote glucose uptake via GLUT4, and ATP adds to insulin action. These effects may be mediated by PI3‐Kγ and Akt and contribute to glucose uptake during muscle activity.Support: FONDAP 15010006, AT‐24100067, CIHR‐MT12601.
The activity of some enzymes in the frog's liver (Rana ridibunda Pall.) in the different natural conditions of thermal water regime
Authors: Bislimi Kemajl Rexhep , Gashi Agim , Bojniku Hazbije , Grapci Linda , Halili Jeton , Mazreku Ilir , Bajraktari Ismet 
Published: June 2021

During the period of March–July 2011, the activity of aspartate aminotranspherase (AST), alanine aminotranspherase (ALT), lactate dehidrogenase (LDH) and alkaline phosphatase (ALP) enzymes was measured in the liver of the Rana ridibunda Pall. natural frog population.Altogether 52 frogs were collected for the research purposes in the two rivers of a different water temperature regime respectively, the thermal water spring of Banja of Malishevë (24.7°C – 29.0°C) and river Tërdec (7.9°C – 19.7°C).There were not recorded great oscillations of AST and ALT activity in the liver of frogs from the thermal water, and ALT activity from the river Tërdec as well during the period of investigation. At the same time in the liver of frogs from the river Tërdec, the lower activity of AST has been recorded in March (12.7±2.6 ), whereas a higher activity in May (41.4±8.4 ), and in July (46.6± 7.6). The distinctive characteristic of this research is that only in March, the activity of AST in the frogs liver from the river Tërdec is significantly at lower level compared with values of frogs from the thermal water.Despite AST, in the liver of the frogs of two localities there were not recorded large oscillations of the ALT, LDH and ALP activity values during the entire period of research.According to the research results we could conclude that the water temperature (the cold stress), the seasonal changes of the environmental temperature, the nutrition, and the metabolic activity of the frogs, affects the activity of AST.