bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lysosomal PPT1‐insufficiency is a common pathogenic link between INCL and JNCL
Authors: Appu Abhilash Puthuvelvippel , Bagh Maria , Sadhukhan Tamal , Mondal Avisek , Mukherjee Anil B. 
Published: April 2020

Neuronal Ceroid Lipofuscinoses (NCLs), also known as Batten disease, constitute a group of the most common inherited childhood neurodegenerative lysosomal storage disorders (LSDs). Even though mutations in >13 different genes cause various NCLs, at the cellular level, all the NCLs characteristically accumulate autofluorescent material (ceroid) in lysosomes and clinically, develop seizures, visual failure and a progressive decline in cognitive and motor functions. These findings suggest shared pathogenic mechanism(s) in these diseases. While inactivating mutations in the CLN1 gene, encoding lysosomal PPT1, cause infantile NCL (INCL), mutations in the CLN3 gene encoding a lysosomal membrane protein of unknown function underlie juvenile NCL (JNCL). PPT1 catalyzes depalmitoylation of S‐acylated proteins, required for their degradation by lysosomal hydrolases. Thus, PPT1‐deficiency causes lysosomal accumulation of S‐acylated proteins (ceroid) leading to INCL. The Cln1−/− and Cln3−/− mice are reliable animal models of INCL and JNCL, respectively. Here we report for the first time that Cln3‐protein requires S‐palmitoylation for its endosomal transport to the lysosomal membrane. Moreover, we found that in the brain of Cln3−/− mice, lysosomal PPT1‐protein as well as Ppt1‐enzymatic activity are significantly lower compared with lysosomes derived from their age‐ and sex‐matched WT counterparts. Unexpectedly, we found that the lysosomes from Cln3−/− mice accumulate significantly higher levels of S‐palmitoylated proteins, characteristically found in those from Cln1−/− mice. Previously, it has been reported that CLN3 mutations suppress the exit of cation‐independent mannose 6‐phosphate receptor (CI‐M6PR) from trans‐Golgi network (TGN), essential for its endosomal trafficking to the late endosome/lysosome. Since CI‐M6PR transports soluble proteins including Ppt1 to the lysosome, we hypothesized that Cln3 mutations may adversely affect the transport of PPT1 to the lysosome causing Ppt1‐insufficiency. Biochemical and confocal microscopic analyses of brain tissues from Cln3−/− mice revealed significantly higher levels of Ppt1 and CI‐M6PR in the Golgi compared to those of their WT counterparts. Collectively, our results demonstrate that lysosomal Ppt1‐insufficiency is a common pathogenic link between INCL and JNCL and suggest that a recently identified Ppt1‐mimetic, neuroprotective small molecule, NtBuHA, may have therapeutic implications for both INCL and JNCL.Support or Funding InformationIntramural Research Program of the Eunice Kennedy Shriver NICHD, NIH
Expression and Immunolocalization of Xenobiotic Transporters at the Human Blood‐Testis Barrier
Authors: Hau Raymond Kin , Wright Stephen H. , Cherrington Nathan J. 
Published: April 2020

The blood‐testis barrier (BTB) is formed by basal tight junctions between adjacent Sertoli cells of the seminiferous tubules and functions as a physical barrier to protect developing germ cells in the adluminal compartment from reproductive toxicants. Multiple xenobiotics are known to cross into the male genital tract (MGT) including male contraceptives, cancer chemotherapeutics, and environmental toxicants, although the mechanism(s) for their movement into the MGT is(are) largely unknown. Previous studies showed that several ATP‐binding cassette (ABC) and solute carrier (SLC) superfamily transporters are expressed in the testes and could potentially facilitate the disposition of xenobiotics into the MGT through concurrent uptake and efflux mechanisms. This study aimed to determine the expression and localization of xenobiotic transporters that may facilitate transepithelial transport across the human BTB. We used immunofluorescence staining to determine the cellular localization of these transporters in the several cell types found in human testes. OAT1, OAT2, OCTN1, and MRP3 were primarily localized to the basal membrane of Sertoli cells, whereas OCTN2, MRP6, and MRP7 were localized to both the basal membrane of Sertoli cells and the peritubular myoid cells that surround the basal membrane of Sertoli cells. Interestingly, CNT2 was localized to the lateral membranes of Sertoli cells and the peritubular myoid cells. OCT1, OCT2, and OCT3 were localized to the peritubular myoid cells and the Leydig cells, although there was moderate staining of spermatogenic cell membranes. However, definitive staining of CNT1, OAT3, MATE1, MATE2, OATP1A2, OATP1B1, OATP1B3, OATP1C1, OATP2A1, OATP2B1, OATP3A1, OATP4A1, and OATP6A1 was not observed due to non‐specific antibody staining or an absence of staining. The localization of CNT2, OAT1, OAT2, OCT1, OCT2, OCT3, OCTN1, OCTN2, MRP3, MRP6, and MRP7 suggests these transporters may contribute to the mechanism(s) by which xenobiotics can cross the BTB. Taken together, the localization of these pharmacologically relevant transporters provides insight into the selectivity of drug disposition across the BTB and may be useful in developing tools to understand and overcome the pharmacokinetic and pharmacodynamic difficulties presented by the BTB.Support or Funding Information5R01GM123643‐045R01GM129777‐02P30CA023074
Retinoic Acid Receptor α Signaling Protects Against Diet‐induced Hepatosteatosis and Obesity in Mice
Authors: Bawa Fathima Nafrisha Cassim , Xu Yanyong , Zhu Yingdong , Chen Shaoru , Gopoju Raja , Zhang Yanqiao 
Published: April 2020

Nonalcoholic fatty liver disease (NAFLD) is often characterized by accumulation of lipids in the liver. It presents a pathological spectrum of changes from simple steatosis to steatohepatitis. It is also often associated with obesity and insulin resistance. Since liver is the main organ involved in retinoid signaling, the impairment of this pathway is shown to increase liver fibrosis with the activation of stellate cells which stores retinoids in liver. However, the role of the retinoid signaling in NAFLD and obesity is not fully understood. Some studies have shown that retinoic acid treatment can ameliorate insulin resistance and obesity. We hypothesize that Retinoic Acid Receptor α (RARα) in hepatocytes play a significant role in mediating retinoid signaling to protect against NAFLD and obesity. To address this hypothesis, we conducted in vivo studies using liver‐specific Rarα knockout mice. Rarα floxed mice were injected with AAV8‐TBG‐Cre or AAV8‐TBG‐Null to generate liver‐specific Rarα knockout mice (L‐Rarα−/−) mice and the control mice (Rarαfl/fl). These mice were gavaged with either vehicle or all‐trans retinoic acid (AtRA; 15mg/kg/day) along with high fat/high cholesterol/high fructose (HFCF) diet for 16 weeks. The data showed that AtRA treatment reduced body fat content and increased energy expenditure in the control mice but not in L‐Rarα−/− mice. In addition, AtRA reduced hepatic triglyceride (TG) levels in both the control mice and L‐Rarα−/− mice with a greater extent in reduction of hepatic TG levels in the control mice. Analysis of hepatic gene expression by qRT‐PCR showed that AtRA reduced hepatic levels of genes involved in lipogenesis and fatty acid uptake (Pparγ, Cd36 and L‐Fabp) or inflammation (Tnf‐α) in the control mice, but not in L‐RARα−/− mice. Similar observations were collected in a different study where control mice and L‐Rara−/− mice were fed an HFCF diet for 20 weeks, followed by 10 weeks of treatment with either vehicle or AtRA. Our data demonstrate that the retinoid signaling protects against the development of hepatosteatosis and obesity in a Rarα‐dependent manner.Support or Funding InformationSupport or funding information: R01DK118805 and R01HL103227
Brazilian Berry Extract, High‐Fat Diet and Prostate Cancer: In Vitro and In Vivo Evaluation of Inflammation and Extracellular Matrix
Authors: Cagnon Valéria H. , Lima Ellen Nogueira , Lamas Celina de Almeida , Baseggio Andressa Mara , Kido Larissa Akemi , Rosseto Isabela Urra , Santos Felipe R. , Montico Fabio , Maróstica Mario Roberto 
Published: April 2020

The high incidence of prostatic cancer (PCa) has encouraged studies in prostatic cellular biology all over the world. In addition, Brazilian berries, such as Myrciaria jaboticaba (Vell.) Berg, show a high polyphenol concentration in the peel, characterizing antioxidative and anti‐inflammatory properties. The aim herein was to analyze the anti‐inflammatory and remodeling tissue effects of the Jaboticaba peel extract (JPE), a patented product, in early prostatic adenocarcinoma development associated with high‐fat diet (HFD) intake, as well as to investigate the JPE role in human PCa and LNCaP cell lines. TRAMP mice from control groups received standard diet or HFD and the treated groups followed the same diet protocol plus JPE (5.8g/Kg/body weight during 5 days per week for 2 months). The dorsolateral prostate was collected for immunohistochemistry and Western Blotting analyses. PC‐3 and LNCaP cells were treated with different doses of JPE (125, 250, 500 ug/mL) and used for cellular viability assay and Western Blotting analyses. The results showed that the HFD intake led to an MMP‐9 increase and a Laminin protein level decrease, worsening the PCa profile in this transgenic model. Conversely, JPE led to decreased MMP‐9 and also TGF‐beta after HFD intake. Laminin level increased in both groups treated with JPE. Dose and time dependent JPE exposure reduced cellular viability, especially, in androgen‐responsive LNCaP cells. In addition, JPE induced NfkB and iNOS reduction not only in LNCaP but also in PC‐3 cell lines, suggesting that the growth inhibitory effect of JPE in PCa may be associated with its anti‐inflammatory role. To conclude, the tissue damage caused by HFD led to the deregulation of the extracellular matrix elements, which intensified cancer progression, compromising the epithelial‐stromal interaction in the dorsolateral prostate. However, JPE was beneficial for the dynamic interaction between the epithelium and stroma in vivo, and altered cancer cell growth by means of anti‐inflammatory pathway, pointing towards a possible adjuvant therapy for PCa delay, highlighting the natural compounds as an important strategy in prostatic disorders.Support or Funding InformationSao Paulo Research Foundation/FAPESP (2018 04579‐7). And Fund for Support to Teaching, Research and Outreach Activities FAEPEX/UNICAMP (2080/20)
Photoreceptor Cell Development Requires CDHR1a Protein Stability Controlled by Siah E3 Ubiquitin Ligase During Zebrafish Eye Development
Authors: Piedade Warlen Pereira , Titialii Kayla , Morris Ann , Famulski Jakub 
Published: April 2020

E3 ubiquitin ligases mediate orderly and precise targeting of protein degradation to maintain biological homeostasis and coordinate proper development. Our lab recently discovered that Siah family of E3 ligases plays a role in early ocular morphogenesis, in particular fusion of the optic fissure. Interestingly, Siah ligases are also expressed during photoreceptor development and are predicted to target CDHR1a, a cadherin superfamily of calcium‐dependent cell adhesion molecules and photoreceptor‐specific cadherin. Mutations in this cadherin are associated with inherited retinal dystrophies, such as cone‐rod dystrophy. Using whole mount in situ hybridization and Immuno‐histochemistry we detected Siah gene expression and protein localization in the outer nuclear layer (ONL) and in the retinal ganglion cell layer starting at 3 days‐post fertilization (dpf) until 7dpf. CDHR1a expression and protein localization overlaps with both Siah genes in the outer segment of the retina from 3 until 7dpf. We therefore hypothesized that Siah regulates CDHR1a during photoreceptor development. To test our hypothesis, I first confirmed siah‐mediated targeting of CDHR1a for degradation using a mammalian cell culture, co‐immunoprecipitation followed by western blotting analysis. Second, I created two transgenic zebrafish lines that express Siah1 or a dominant negative (Siah1DRING) under the control of the heat shock promoter. Using the heat shock line to overexpress Siah1 at 2dpf and 2.5dpf, we observed a decrease in the number of developing rods and cones in the retina at 3dpf. The number of retinal ganglion cells, amacrine, bipolar and horizontal cells, however, did not change. In addition, there was a significant reduction of proliferating (EdU+ and PH3+ cells in ONL) cells in the retina. Taken together, our results suggest that Siah ubiquitin ligases may control CDHR1a stability and therefore regulate the photoreceptor cell proliferation and differentiation.Support or Funding InformationThis work was supported by the Brazilian National Council for Scientific and Technological Development (CNPq) under grant number 202970/2014‐0.
Four‐week Ketogenic Diet Improves Aerobic Metabolic Capacity in Extensor Digitrum Longus Muscle of Sedentary Male Rats
Authors: Ogura Yuji , Yoshihara Toshinori , Kurosaka Mitsutoshi , Kakigi Ryo , Fujiwara Sei-etsu , Kakehashi Chiaki , Higashida Kazuhiko , Funabashi Toshiya 
Published: April 2020

Ketogenic diet is an extremely high‐fat diet with low or absence of carbohydrate in the total nutrient. Thus, this diet causes ketone body production from the liver and shifts energy sources from carbohydrate to fat. Whereas previous studies have suggested that ketogenic diet ameliorates brain disorder such as drug‐resistant epilepsy, its influence on peripheral tissue is unclear. Skeletal muscle is an important organ for locomotion and metabolism. Recent studies have suggested that the ketogenic diet changes the energy metabolic properties in the skeletal muscle as well; however, the results are contradictory. Moreover, it remains unknown whether the ketogenic diet affects the muscle contractile function. Therefore, this study aimed to clarify the effect of ketogenic diet on muscle performance and biochemical adaptation in rat skeletal muscle. This study was approved by Institute for Animal Experimentation at St. Marianna University School of Medicine. Male Wistar rats (8 weeks of age) were assigned to either control group (CON) or ketogenic diet group (KD). CON and KD were fed for 4 weeks either control diet (10% fat, 10% protein, 80% carbohydrate) or ketogenic diet (90% fat, 10% protein, 0% carbohydrate), respectively. Body mass was recorded once a week. After the 4‐week feeding period, the animals were anesthetized, and the blood sample was collected to analyze blood ketone concentration. The right extensor digitrum longus (EDL) muscle was used to evaluate the twitch force, tetanic force, and fatigue ex vivo. The left EDL was homogenized and used to analyze the myosin heavy chain composition, the protein expression of metabolic enzymes, and the citrate synthase activity. Body mass was lower in KD than CON at third and fourth week of the feeding period. Blood ketone concentration in KD was significantly higher than CON. There was no statistical difference in the twitch and tetanic forces as well as muscle fatigue between CON and KD. However, the citrate synthase activity in KD was significantly higher than CON. The protein expression of citrate synthase, succinate dehydrogenase, cytochrome c oxidase subunit 4, and 3‐hydroxyacyl‐CoA dehydrogenase in KD was significantly higher than CON. Moreover, there was a shift of myosin heavy chain from type IIb to IIx in KD. Therefore, we conclude that 4‐week ketogenic diet improves skeletal muscle aerobic capacity without obstructing muscle contractile function in the sedentary male rats.Support or Funding InformationThis work was supported by MEXT/JSPS KAKENHI Grant Numbers JP24650334, JP18K17767.
The Application of 3D Printing and Nutrient/Biomaterial Microhabitats for In Situ Enrichment of Microbial Cultures
Authors: Andersen Morten , Moreira-Grez Benjamin , Andersen Magdalena Julia , Whiteley Andrew 
Published: April 2020

The objective of this study was to develop a new tool for selectively capturing new microorganisms from the environment. High‐throughput environmental sequencing has revealed a diversity much greater than that which we have brought into culture. Even so, the microorganisms that have been successfully cultured have yielded a wealth of pharmaceuticals, enzymes and living cultures for medical, agricultural and industrial use. Bringing new microorganisms into culture thus has great value.A MicroCaptureArray for high‐throughput in situ enrichment of new species from the environment using metabolic cues was created using 3D printing. The system is based on repurposing biomaterials developed for medical implants and controlled release devices
[1],
[2]. The MicroCaptureArray was tested in a terrestrial (soil) and an aquatic (pond) environment for 9 and 7 days, respectively. DNA was then isolated from the array and subjected to 16S sequencing with Qiime2 data analysis.The MicroCaptureArray attracted, captured and enriched different species, many of which were undetectable in the background community using 16S sequencing. The different metabolic cues resulted in the enrichment of different microorganisms. The system also enriched potentially new species, in several samples more then 25% of the captured microorganisms were unassignable beyond the Kingdom of Bacteria or the Phylum of Proteobacteria, in constrast, unassignable bacteria accounted for less than 0.3% of the background samples.In conclusion, the MicroCaptureArray shows that metabolic cues may be used for in situ baiting and enrichment culture of bacteria. The tool may allow researchers to detect rare microorganisms that are present but undetectable in the environment that may nonetheless play a major role when certain nutrients are introduced. It may also allow selective attraction and enrichment of environmental bacteria that are capable of metabolizing particular substances. This may allow targeted discovery of entirely new species with desirable properties.Support or Funding InformationThe project was supported by the Villum Foundation (grant number 17627)
Tabata style functional exercise increases resting and postprandial fat oxidation but does not reduce triglyceride concentrations
Authors: Pearson Regis , Green Edward S. , Olenick Alyssa A. , Jenkins Nathan T. 
Published: April 2020

We evaluated the effect of a high fat meal with and without prior high intensity functional exercise executed in a Tabata style interval pattern on resting and postprandial substrate oxidation, to include blood glucose and triglyceride concentrations in humans. Eleven healthy males completed two trials [Tabata exercise (TE) and non‐exercise control (CON)] in random order separated by 7 days. A two‐day protocol was used in which TE or CON was performed on the first day and a high fat meal was administered ~13 hrs later on the following morning. Power output from the TE session was quantified using a kinematic approach by calculating external work performed per unit of time for each of the 4 exercises (rowing, dumbbell thrusters, kettlebell swings, and burpees). For the meal challenge, respiratory gases and blood samples were taken fasted and at 1, 2, and 3 hrs postprandial. Fat oxidation was significantly higher after TE compared to CON at each time point (P < 0.05). Total fat oxidized, total fat oxidation AUC, and total postprandial fat oxidized all significantly increased after TE (P < 0.05). Carbohydrate oxidation was significantly lower after TE compared to CON at 1 hr postprandial (P < 0.05). Total carbohydrate oxidized, total carbohydrate oxidation AUC, and total postprandial carbohydrate oxidized all significantly decreased after TE (P < 0.05). There were no significant effects of TE on fasting or postprandial glucose or triglyceride concentrations. Functional exercises performed in a high intensity TE pattern enhance fasting and postprandial fat oxidation on the following day with minimal influence on blood triglycerides or glucose levels.Support or Funding InformationR.C.P. was support by the University of Georgia Graduate School Georgia Research Education Award Traineeship Follows Program
Quantifying Transcription Dynamics of CUP1 gene of S. cerevisiae at the Single‐Molecule Level
Authors: Mehta Gunjan D. , Ball David , Karpova Tatiana 
Published: April 2020

Transcription factors (TFs) show rapid and reversible binding to gene promoters to activate transcription in live cells. Transcription often occurs in sporadic bursts, however it is yet poorly understood how the dynamics binding of TFs is coordinated with gene bursting. Gene promoters are generally occupied by the nucleosomes and it is known that the presence of a nucleosome at the gene promoter reduces the accessibility for TF binding. Additionally, nucleosomes are known to evict the TFs from the gene promoters. Hence, chromatin remodeling plays an instrumental role in modulating TF binding. Due to advancements in the Single‐Molecule Imaging approaches, quantification of the transcription parameters such as initiation rate, transcript production time, burst frequency, burst duration and burst size has become achievable. We developed and employed several Single‐Molecule approaches (Single‐Molecule Tracking (SMT), live imaging of CUP1 transcription site (TS) by PP7 stem‐loop approach and smFISH modeling of CUP1 transcription) to quantify the transcription dynamics of the CUP1 gene of S. cerevisiae. In cell population, we observe slow sequential cycles of CUP1 transcription in conditions of the prolonged heavy metal stress. By applying gene bursting model, we extract the dynamics of the transcription parameters within the slow cycle from live TS and smFISH data and correlate these parameters with the dynamics of TF and POLII. The chromatin remodeler RSC speeds up the search process of the TF Ace1p for its Response Elements (RE) at the CUP1 promoter. RSC chromatin remodeler also affects the transcription of CUP1. We investigate the effects of RSC remodeler on the slow cycle of CUP1. Modulation of burst frequency yields more effective and productive response than the modulation of burst amplitude. We are interested in cyclical expression of the genes in stress response.Support or Funding InformationThis work was supported by Intramural Research Program of the National Institutes of Health (NCI, CCR).
Effects of [6]‐Gingerol on β‐amyloid‐induced Cognitive Impairment in Mice
Authors: Bae Hui Jin , Mun Seul Ah , Jang Jung-Hee , Park Gyu Hwan 
Published: April 2020

Alzheimer’s disease (AD) is a neurodegenerative disease characterized by disruption of cholinergic neurons and it is likely that promoted inflammation and oxidation is involved in pathogenesis of AD. Consequently, it is worthwhile developing effective phytochemical agents which can reinforce the cholinergic transmission and anti‐inflammatory defense capacity for the prevention and/or treatment of AD. In this study, we tested whether [6]‐gingerol, an active constituent of ginger could reduce β‐amyloid (Aβ)‐induced cognitive impairment in mice. Administration of [6]‐gingerol p.o. (25 mg/kg) significantly alleviated Aβ‐induced cognitive dysfunction as evaluated by Y‐maze and passive avoidance tests. To further elucidate the mechanisms underlying the effects of [6]‐gingerol, the possible involvement of inflammatory mediators were examined. Treatment of [6]‐gingerol reduced the expression of inflammatory enzymes and pro‐inflammatory cytokines such as COX2, IL‐1β, TNFα and iNOS. In addition, [6]‐gingerol decreased the phosphorylation of p66‐Shc which had been increased by Aβ. Taken together these findings suggest that [6]‐gingerol might potentiate anti‐inflammatory cellular defense system and have therapeutic potentials in the management of cognitive impairment in AD.Support or Funding InformationThis work was supported by the National Research Foundation of Korea (NFR) grant funded by the Korea government (MSIT)