bluuebunny/crossref_metadata_2025 huggingface dataset:
Ringing the universe with cosmic emptiness: void properties through a combined analysis of stacked weak gravitational and Doppler lensing
Authors: Hossen Md Rasel , Ema Sonia Akter , Bolejko Krzysztof , Lewis Geraint F
Published: May 2022

An essential aspect of cosmic voids is that these underdense regions provide complementary information about the properties of our Universe. Unlike dense regions, voids are avoided by matter and are less contaminated by baryonic processes. The first step to understanding the properties of cosmic voids is to correctly infer their mass profiles. In the literature, various techniques have been implemented. In this paper, we review them and implement a new technique that is based on Doppler lensing. We use a relativistic N-body code, Gevolution, to generate cosmological mass perturbations and implement a three-dimensional (3D) ray-tracing technique that follows the evolution of ray-bundles. We focus on the various properties of cosmic voids (e.g. void size function, 2-point correlation function, and the density profile of voids), and compare the results with their universal trends. We show that when weak lensing is combined with the Doppler lensing we obtain even tighter constraints than weak lensing alone. We also obtain better agreement between density profiles within central parts of voids inferred from lensing and density profiles inferred from halo tracers. The implication of the result relevant to the ongoing and prospective low-redshift spectroscopic surveys is briefly discussed.
Multidisciplinary management of chronic refractory pain in autosomal dominant polycystic kidney disease
Authors: van Luijk Franka , Gansevoort Ron T , Blokzijl Hans , Groen Gerbrand J , de Haas Robbert J , Leliveld Anna M , Meijer Esther , Perdok Joke M , Stellema Ruud , Wolff Andreas P , Casteleijn Niek F
Published: May 2022

Background

Chronic pain is often difficult to manage in autosomal dominant polycystic kidney disease (ADPKD) patients and sometimes even leads to nephrectomy. We analyzed the long-term efficacy of our innovative multidisciplinary protocol to treat chronic refractory pain that aims to preserve kidney function by applying among other sequential nerve blocks.

Methods

Patients were eligible if pain was present ≥3 months with a score of ≥50 on a visual analog scale (VAS) of 100, was negatively affecting quality of life and if there had been insufficient response to previous therapies, including opioid treatment. Treatment options were, in order, analgesics, cyst aspiration and fenestration, nerve blocks and nephrectomy.

Results

A total of 101 patients were assessed in our clinic (mean age 50 ± 11 years, 65.3% females). Eight patients were treated with medication, 6 by cyst aspiration or fenestration, 63 by nerve blocks and 6 received surgery as the first treatment option. Overall, 76.9% experienced a positive effect on pain complaints shortly after treatment. The VAS score was reduced from 60/100 to 20/100 (P < 0.001) and patients decreased their number of nonopioid and opioid analgesics significantly (P < 0.001, P = 0.01, respectively). A substantial number of the patients (n = 51) needed additional treatment. At the end of follow-up in only 13 patients (12.9%) was surgical intervention necessary: 11 nephrectomies (of which 10 were in patients already on kidney function replacement treatment), 1 liver transplantation and 1 partial hepatectomy. After a median follow-up of 4.5 years (interquartile range 2.5–5.3), 69.0% of the patients still had fewer pain complaints.

Conclusions

These data indicate that our multidisciplinary treatment protocol appears effective in reducing pain in the majority of patients with chronic refractory pain, while postponing or even avoiding in most patients surgical interventions such as nephrectomy in most patients.
Transition in Thermal History and Recurring Burial-Exhumation Cycles along Colder Thermal Gradients at the Archaean-Proterozoic Boundary: New Insights from the Western Dharwar Craton, South India
Authors: Dasgupta Anamitra , Bhowmik Santanu Kumar , Dasgupta Somnath
Published: May 2022

The Hole Narsipur Greenstone Belt (HGB), in the Western Dharwar Craton, South India, preserves evidence for three cycles of metamorphism spanning the Archaean–Proterozoic transition. Changes in metamorphic style across the transition are investigated using integrated petrographic, mineral chemical, metamorphic evolutionary history, conventional geothermobarometry, and phase equilibria modeling studies of rocks of diverse bulk rock composition from the belt. These rocks record signatures of three overprinting metamorphic cycles (M2 → M3 → M4 in sequence), postdating a Mesoarchaean, M1 event in the craton. The M2 cycle represents characteristic LP–HT metamorphism along a counterclockwise (CCW) metamorphic P–T path, peak metamorphism at 4–5 kbar, 515–595°C, and high thermobaric ratios (T/P ~1050–1525°C/GPa) at the M2 peak. The intermediate M3 metamorphic cycle records a prograde burial of partially cooled M2 rocks to middle-lower crustal depths, peak metamorphism at P ~7.5–10.2 kbar, T ~630–655°C, and corresponding intermediate T/P ratios (T/P ~620–875°C/GPa) and a retrograde stage, marked by combined exhumation and cooling to P ~5.3–6.4 kbar, T ~530–575°C. The resultant P–T paths of evolution show variability of clockwise (CW) paths from classical hairpin type to two-stage prograde heating segments. During the M4 metamorphism (TMax ~630–640°C at ~8.9–10.3 kbar and with lower T/P ratios at ~610–720°C/GPa), the partially exhumed M3 crust is re-buried to lower crustal depths along an intermediate thermal gradient as during the M3 metamorphic cycle. We interpret the thermal transition, tectonic thickening along a cooler thermal gradient, repeated burial-exhumation cycles in the orogenic wedge, and tectonic mixing of the three metamorphic cycles in terms of a three-stage tectonic evolution of the HGB. These in a sequence are (a) the development of a hot and thinned crust (cf. M2 cycle), (b) the transition to a thickened crustal zone tectonic domain (cf. M3 cycle), with both the domains being part of the lithospheric peel-back driven convergence setting and (c) finally continental collision tectonics (cf. M4 cycle). The pulsating nature of orogenesis at the Archaean–Proterozoic boundary deduced here distinguishes the Western Dharwar Craton from other cratonic blocks globally at the same time frame.
Affordable Housing and City Welfare
Authors: Favilukis Jack , Mabille Pierre , Van Nieuwerburgh Stijn
Published: May 2022

Housing affordability is the main policy challenge for most large cities in the world. Zoning changes, rent control, housing vouchers, and tax credits are the main levers employed by policymakers. How effective are they at combatting the affordability crisis? We build a dynamic stochastic spatial equilibrium model to evaluate the effect of these policies on the well-being of its citizens. The model endogenizes house prices, rents, construction, labour supply, output, income, and wealth inequality, the location decisions of households within the city as well as inter-city migration. Its main novel features are risk, risk aversion, and incomplete risk-sharing. We calibrate the model to the New York metropolitan statistical area. Housing affordability policies carry substantial insurance value but affect aggregate housing and labour supply and cause misallocation in labour and housing markets. Housing affordability policies that enhance access to this insurance especially for the neediest households create substantial net welfare gains.
Racial Diversity and Racial Policy Preferences: The Great Migration and Civil Rights
Authors: Calderon Alvaro , Fouka Vasiliki , Tabellini Marco
Published: May 2022

Between 1940 and 1970, more than 4 million African Americans moved from the South to the North of the US, during the Second Great Migration. This same period witnessed the struggle and eventual success of the civil rights movement in ending institutionalized racial discrimination. This article shows that the Great Migration and support for civil rights are causally linked. Predicting Black inflows with a shift-share instrument, we find that the Great Migration raised support for the Democratic Party, increased Congress members’ propensity to promote civil rights legislation, and encouraged pro-civil rights activism outside the US South. We provide different pieces of evidence that support for civil rights was not confined to the Black electorate but was also shared by segments of the white population.
Private foundations in the Czech Republic: 10 years on the road
Authors: Ronovská Kateřina , Pihera Vlastimil
Published: May 2022

In the early years following the enactment of the “new” Czech Civil Code it was not clear whether the newly introduced conceptual approaches in the area of foundation law would prove successful. The “private element”, or the possibility to use these structures as a tool for managing (family) assets, has not only brought considerable dynamics to the foundation sphere, but also the need for a fundamental change in thinking.

In 2021, it is a reality that foundations (foundations and foundation funds), in addition to trust funds, are beginning to be used for “private” purposes.

The authors of this article aim to outline the basic contours of the development and the current state of affairs of private foundations and foundation funds in the Czech legal environment.
Second Year Medical Students Learning by Teaching Pregnant Women About Physiological Changes During Pregnancy
Authors: Shrestha Bipin K. , Thapa Bikalp , Shrestha Rashmi , Amatya Tara Man , Khatri Ratna
Published: May 2022

Classroom‐based lectures are commonly used for teaching Basic sciences in many medical schools. It is difficult to keep the students motivated to learn with only didactic lectures during the preclinical years. We need to constantly explore teaching methods to keep the learning enthusiasm consistent, improve understanding and enhance students’ performance. Studies are important to prove students' achievement will increase when active methodologies are properly applied, either in isolation or in conjunction with lectures. This study aimed to assess the performance and perceptions of Second‐year medical students who were put through an activity that included the involvement of students in educating pregnant women on physiological changes during pregnancy. At first, the students completed a pretest after the lectures on the physiology of pregnancy. Next, the students visited the Ante Natal Care Out Patient Department (ANC OPD) and educated the pregnant mothers using various teaching tools like flip charts, models, pamphlets and diagrams prepared by themselves. Then completed a posttest. In addition, feedback regarding this teaching‐learning approach was obtained from the students. The results showed that the mean (SD) scores after the activity were greater compared to before (MCQs, P‐Value 0.01, True/False questions 0.001 and combined total, <0.001). From the feedback, we found that 94.11% of the students improved their understanding of the physiology of pregnancy, 85.28% of students were positive about the motivation to learn the physiology of pregnancy and 94.2% of the students were eager to have similar activities in the future. Therefore, this active learning method not only improved understanding of the physiological concept but also increased the performance of the students and provided motivation to learn. Such learning strategies can be applied in other Biological and Biomedical sciences embracing relevant clinical scenarios.
Integrating Basic and Clinical Sciences for Medical Undergraduate Students to Enrich Learning
Authors: Peck Gina , Doroudi Majid , Wallace Clarissa , Almehthel Mohammed , Fejtek Monika
Published: May 2022

Background

Medical students come from increasingly diverse backgrounds; this diversity reflects the populations students will eventually serve and to which medical education adapts. In 2015, the MD Undergraduate Program at the University of British Columbia (UBC) implemented a spiral curriculum emphasizing the integration of basic and clinical sciences from the first year of medical education. For students who come from non‐science backgrounds, the immediate dive into both basic and clinical science can be overwhelming. Enrichment materials, such as interactive modules, provide alternative and additional means of understanding and applying curriculum content. The purpose of this project was to develop an interactive module to supplement the topic of adrenal dysfunction by presenting clinical scenarios in the context of adrenal gland physiology.

Methods Used

Based on curriculum content, an interactive module combining the physiology of the adrenal gland with two different clinical scenarios on adrenal dysfunction was developed. This module was made available to all undergraduates of the UBC MD Program, but was specifically designed for second year students during their week‐long topic on adrenal dysfunction. At the end of the module, an optional survey was provided to assess the module’s design and content and to determine if similar modules would be desired.

Summary of Results

Upon completion of the module, all participants rated their knowledge of the adrenal cortex as Strong (66%) or Very Strong (33%), while knowledge of the two clinical scenarios was split evenly among students into Very Strong, Strong, and Average. 75% of students wished to see similar modules for curricular content, and all students felt that the module was an effective resource for the week’s content.

In terms of design, 66% of participants found the study resources most useful, and all participants felt that the cases demonstrated clinical presentations and management better than average: Extremely Well (66%) and Very Well (33%) in terms of clinical presentation; and Extremely Well (33%) and Very Well (66%) in terms of management. Finally, 66% of students felt that the module did Extremely Well in terms of the integration of basic and clinical science while 33% felt it did Moderately Well.

Conclusion

An interactive module overall improved students’ subjective knowledge of the basic and clinical science of adrenal dysfunction. The majority of students were most appreciative of study resources and were in favour of similar enrichment modules for other curricular content.
Differential Expression of Nuclear Body Component, Zc3h8, Disrupts DNA Double Strand Break Repair
Authors: Schmidt John A. , Sato Rhiann
Published: May 2022

The processes of DNA damage recognition and repair are paramount to maintaining genome integrity in the cell. Failure to do so results in genetic instability and the accumulation of mutations. Double strand breaks have the greatest potential to introduce mutations because rearrangements, non‐homologous end joining, deletions, or insertions may result. Unrepaired or incorrectly repaired DNA double strand breaks can lead to cell death, apoptosis, or disease such as cancer. We previously demonstrated that the C3H1 zinc finger‐containing protein Zc3h8 (also called Fliz1) promotes oncogenic phenotypes when overexpressed in cells, while oncogenic phenotypes are suppressed when expression is reduced. In mice, Zc3h8 expression affected tumorigenesis and tumor size in vivo. Higher expression of Zc3h8 led to more frequent tumors and larger tumors in mice. In humans Zc3h8 is overexpressed in 2‐6% of cancers and is a statistically significant prognostic indicator for poor rates of survival. The Zc3h8 protein has been localized to nuclear bodies including promyelocytic leukemia (PML) and Cajal Bodies where key cellular processes including transcription, translation, post‐translational modifications, and RNA processing are regulated. PML Bodies, in particular, have a role in DNA damage response and repair since P53 is located in PML Bodies along with DAXX, ATRX, and CK2. Since Zc3h8 co‐localized with DNA damage response and repair proteins in PML bodies and promotes oncogenic properties in cells, we investigated if Zc3h8 expression alters how cells respond to DNA damage such as double strand breaks and how this damage is mitigated in cells. Differential expression of Zc3h8 affected mouse mammary tumor cell’s ability to repair double strand breaks caused by the topoisomerase II inhibitor etoposide as indicated by the presence of γH2AX, a marker for unrepaired double strand breaks. Using a Comet assay, we found extensive DNA fragmentation in cells with elevated Zc3h8 expression despite high rates of proliferation and migration. Furthermore, Zc3h8 increases cellular tolerance for DNA damage and alters apoptosis. These phenotypes promote further mutation and may indicate a molecular cause for highly aggressive and progressive cancers with elevated Zc3h8 expression. These conclusions suggest a DNA damage repair and response pathway that is altered by Zc3h8 expression levels can affect multiple aspects of cellular behavior and promote oncogenic phenotypes.
Nanometers matter in immune evasion mediated by the cellular glycocalyx
Authors: Paszek Matthew J.
Published: May 2022

Cancer cells construct a glycocalyx with biochemical and physical attributes that protect against immune surveillance. While it has been proposed that the glycocalyx may form a nanoscale‐thick barrier against immune cell attack, testing this possibility has proven difficult due to the challenges associated with imaging the ultrathin glycocalyx structure, particularly in live cells. In this talk, I will discuss how an imaging technique called Scanning Angle Interference Microscopy (SAIM) can be applied to accurately measure the nanoscale thickness of the cellular glycocalyx. Using SAIM and glycoengineering strategies, we reveal how the surface density, glycosylation, and crosslinking of cancer‐associated mucins contribute to the nanoscale material thickness of the glycocalyx, and further analyze the effect of the glycocalyx thickness on resistance to effector cell attack. Natural Killer (NK) cell‐mediated cytotoxicity exhibits a near perfect inverse correlation with the glycocalyx thickness of target cells regardless of the specific glycan structures present. Changes in glycocalyx thickness as small as 10 nanometers can significantly alter susceptibility to immune cell attack. Finally, I will discuss strategies for overcoming the glycocalyx barrier through cellular engineering of immune cells. These strategies include the surface display of glycocalyx‐editing enzymes on the NK surface for improved penetration of the glycocalyx barrier.
The Vitamin K2Synthetic Enzyme UBIAD1 Moonlights as a Key Regulator of Cholesterol Synthesis
Authors: DeBose‐Boyd Russell A. , Jo Youngah , Schumacher Marc M.
Published: May 2022

UbiA prenyltransferase domain‐containing protein‐1 (UBIAD1) utilizes geranylgeranyl pyrophosphate (GGpp) to synthesize the vitamin K2 subtype menaquinone‐4 (MK‐4). Mutations in UBIAD1 cause Schnyder corneal dystrophy (SCD), which is characterized by corneal opacification owing to over‐accumulation of cholesterol. Our studies disclosed a key role for UBIAD1 in regulating endoplasmic reticulum (ER)‐localized HMG CoA reductase, the rate‐limiting enzyme in synthesis of cholesterol and nonsterol isoprenoids including GGpp. Feedback control of reductase involves sterol‐induced ubiquitination, an obligatory reaction for its ER‐associated degradation (ERAD) that is augmented by GGpp. Sterols also cause UBIAD1 to bind reductase, which inhibits ERAD and allows continued synthesis of nonsterol isoprenoids in sterol‐replete cells. GGpp triggers release of reductase from UBIAD1, enhancing ERAD and stimulating translocation of UBIAD1 to Golgi. SCD‐associated UBIAD1 resists GGpp‐induced release from reductase and becomes sequestered in ER to inhibit ERAD.

Gene knockout studies in mice were attempted to explore the in vivo function of UBIAD1; however, homozygous germ‐line deletion of Ubiad1 caused embryonic lethality. We generated homozygous deletion of Ubiad1 in knock‐in mice expressing ubiquitination‐resistant HMGCR, implying embryonic lethality results from enhanced ERAD of HMGCR. The study of Ubiad1‐deficient mice offers the opportunity to determine the physiological significance of UBIAD1‐mediated synthesis of MK‐4.
Re‐Framing Reality: Framework Analysis as a Step‐Wise Approach to Thematic Analysis
Authors: Lazarus Michelle
Published: May 2022

Similar to quantitative methods, there are many methods researchers can use for analysing qualitative research. This talk will introduce Framework Analysis, a systematic qualitative method often used in healthcare education research. Framework Analysis can be used in both cross‐sectional, and longitudinal qualitative research, and consists of the following five steps: Data familiarization; thematic framework identification; indexing; charting; mapping and interpretation. Participants in this session will be introduced to qualitative language (and related meanings), as well as opportunities to practice the practical processing of each step. Each participant will be supported to engage in the Framework analysis steps by working through relatable examples alongside their audience peers. This session will be highly interactive. By the end of this session, participants will be more familiar with Framework analysis language and process, and are likely to have met a new colleague through this activity.
Native American and First Nation Perspectives on the Use of Human Remains in Education & Proximity to Death
Authors: Reynolds Amberly M. , Pennefather‐O’Brien Elizabeth
Published: May 2022

Cultural attitudes regarding interactions with the deceased, the dead and contact with the lifeless human body vary throughout the world, including among Native American, First Nation, Metίs, and Inuit groups in North America. From remains that have been obtained without consent to working with cadaveric specimens, this talk addresses the beliefs and concerns of indigenous peoples regarding the use of human remains as educational specimens and interaction with the dead. Both speaker perspectives as well as preliminary results from research will address the diverse attitudes towards the dead, treatment of the dead, use of human remains for teaching and research purposes. We will also acknowledge the principle of informed consent in a colonial world. Anatomical education is often reliant on the gracious donation of humans but what are we to do if they were not obtained with consent as are many of the indigenous specimens still maintained at universities?

Working with human remains can also be of concern for indigenous students with respect to their cultural beliefs. This talk will address the importance and considerations needed for indigenous students while providing the audience with a basic understanding of some of those cultural beliefs. It is not only important to have respect for the human remains but also for the students who encounter them. Anatomical educators must have an awareness of cultural and religious beliefs that are at odds with working with human remains or being in the presence of a lifeless body, so that we are prepared to provide inclusive learning environments for our students. We hope to provide an inclusive discussion on Native American and First Nation perspectives on the use of human remains in education and the proximity to death.
Fascia in Myofascial Pain Syndrome
Authors: Stecco Antonio
Published: May 2022

Myofascial pain syndrome (MPS) is described as the muscle, sensory, motor, and autonomic nervous system symptoms caused by stimulation of myofascial trigger points (MTP), but the participation of fascia in this syndrome has often been neglected. This presentation will explore the important contribution of structural and functional alterations of the deep fascia to the pathophysiology and clinical manifestation of MPS. The interaction of the three dimensional fascial system, with the dynamic nature of sensitization, will be presented in the context of the pathophysiology of MTP and the expression of quantitative, reproducible physical findings in the clinical manifestation of chronic myofascial pain. Emphasis will be given to recent developments in the biology of fascia, and in particular, its hyaluronan (HA)‐rich matrix content that provide a viscoelastic property possible cause of MPS. The presentation will translate these findings to highlight novel clinical directions in the diagnosis and non‐pharmaceutical management of MPS.
Ca2+‐activated Sphingomyelin Scrambling and Turnover Mediate ESCRT‐independent Lysosomal Repair
Authors: Holthuis Joost , Niekamp Patrick
Published: May 2022

Lysosomes are vital organelles vulnerable to injuries from diverse materials. Failure to repair or sequester damaged lysosomes poses a threat to cell viability. Here we report that cells exploit a sphingomyelin‐based lysosomal repair pathway that operates independently of ESCRT to reverse potentially lethal membrane damage. Various conditions perturbing organelle integrity trigger a rapid calcium‐activated scrambling and cytosolic exposure of sphingomyelin. Subsequent metabolic conversion of sphingomyelin by neutral sphingomyelinases on the cytosolic surface of injured lysosomes promotes their repair, also when ESCRT function is compromised. Conversely, blocking turnover of cytosolic sphingomyelin renders cells more sensitive to lysosome‐damaging drugs. Our data indicate that calcium‐activated scramblases, sphingomyelin, and neutral sphingomyelinases are core components of a previously unrecognized membrane restoration pathway by which cells preserve the functional integrity of lysosomes.
The combination of NRF1 and Nrf2 activators in myoblasts stimulate mechanisms of proteostasis without changes in mitochondrial respiration
Authors: Walsh Maureen A. , Zhang Qian , Musci Robert V. , Hamilton Karyn L.
Published: May 2022

Persistent oxidative stress contributes to hallmarks of aging, including impaired proteostasis and mitochondrial dysfunction, while acute oxidative challenges resolved swiftly contribute to beneficial adaptations. Adaptive homeostasis is where acute exposures to sub‐toxic stimuli kindle transient expansion of responses necessary to reestablish homeostasis. Elucidating mechanisms underlying adaptive homeostasis will provide novel targets for healthspan extension. Nrf2 is a key regulator of cytoprotective gene transcription for redox homeostasis; NRF1 is a transcription factor that regulates expression of genes necessary for mitochondrial function. Regulation of both is compromised with advancing age. We hypothesized that NRF1 (NRF1a) and Nrf2 (Nrf2a) activators might improve adaptive homeostasis in response to an oxidative challenge by promoting mitochondrial proteome maintenance and function in C2C12 myoblasts. Using deuterium stable isotope tracing, we assessed protein synthesis over a 16‐hr treatment with a CON (DMSO), NRF1a, Nrf2a, or both, with and without a hydrogen peroxide (H2O2) stress. We assessed mitochondrial function using high‐resolution respirometry. Co‐treatment of NRF1a and Nrf2a under H2O2 stress favored proteostatic maintenance (p<0.05) through maintained protein synthesis (p>0.05), but a decrease in cell proliferation (p<0.05). H2O2 stress significantly decreased mitochondrial maximal respiration, but this decrease was not rescued by NRF1a/Nrf2a co‐treatment indicating the same ATP availability. Interestingly, there were no differences in submaximal ADP stimulated respiration or ADP sensitivity between CON, H2O2, or the NRF1a/Nrf2a co‐treatment (p>0.05). Further, there were no differences in endogenous redox capacity or mitochondrial protein content between treatment groups. These results suggest that simultaneously targeting NRF1 and Nrf2 may be a viable approach for reestablishing mitochondrial protein homeostasis following a stress, but that this adaptation may not improve respiratory capacity.
A novel NSC small molecule inhibitor inhibits proliferation of triple‐negative breast cancer cells through metabolic reprograming
Authors: Yousefi Hassan , Lauterboeck Lothar , Okpechi Samuel , Zabaleta Jovanny , Guidry Jessie , Yang Qinglin , Alahari Suresh
Published: May 2022

Background

Treatment of patients with triple‐negative breast cancer (TNBC) has been challenging due to the absence of well‐defined molecular targets and high invasive and proliferative capacities of these cells. Current treatments against TNBC have shown minimal activity due to the high recurrence rate in the patients. There is, therefore, a pressing need for novel and efficacious therapies against TNBC. Here, we found a novel small molecule inhibitor with potent anti‐tumor activity against TNBC cells.

Methods

Anti‐proliferative effects of NSC small molecule inhibitor were determined using 2D and 3D culture cell proliferation assays. Using proteomics, NGS and enrichment analysis, we globally investigated top regulatory pathways affected by this compound in TNBC cells. Subsequently, we validated the proteomics and NGS analysis data using seahorse and enzymatic assays. Finally, we showed the inhibitor anti‐tumor effects in vivo. Data were evaluated in triplicate against untreated control cells. Data were graphed and analyzed using GraphPad Prism Software 8.0 using one‐way ANOVA and the unpaired two‐tailed Student’s t‐test.

Results

In this report, we showed that a novel NSC small molecule inhibitor reduced the proliferation of TNBC cells. Proteomic analysis confirmed a significant metabolic reprograming including suppression of glycolysis and oxidative phosphorylation after treatment. Furthermore, we found that treatment with NSC small molecule inhibitor impaired glycolysis and oxidative phosphorylation via modulating the activity of metabolic regulator enzymes.

Conclusions

Altogether, our data indicate that NSC small molecule inhibitor may exhibit anti‐tumor activity in TNBC cells and provide a rationale for further investigation of the potential of NSC small molecule inhibitor as an attractive therapeutic drug for TNBC. Further in‐depth investigations are required to evaluate the exact targets and mechanism of this potent small molecule inhibitor, and its anti‐neoplasm effects on TNBC in vivo.
Novel Chemical Probes targeting the Polyamine Regulatory Circuit
Authors: Tulluri Vennela , Suckiel Michaela , Acevedo Luis , Lyness Conor , Browne Amanda C. , Nemmara Venkatesh
Published: May 2022

Polyamines are vital for cellular life. Polyamines such as putrescine, spermidine, and spermine regulate the essential cellular process, including gene expression and cell proliferation. Given its cellular role, polyamine concentration in cells is tightly controlled. However, aberrant polyamine metabolism is associated with numerous diseases, including cancer. Growing evidence suggests that elevated polyamine levels are the absolute requirement for tumor growth and progression. The concentrations of cellular polyamines are controlled by the polyamine regulatory circuit comprising three different proteins: Ornithine Decarboxylase (ODC), Ornithine Decarboxylase Antizyme (OAZ), and Antizyme Inhibitor (AZIN). While ODC is directly involved in polyamine biosynthesis, OAZ and AZIN regulate the ODC activity via protein‐protein interactions. The dysregulation of ODC, OAZ, and AZIN leads to elevated polyamines in numerous pathologies, making them attractive targets to control polyamine levels. Besides regulating polyamine synthesis, OAZ is believed to modulate polyamine transport via multiple pathways. However, the precise mechanism of OAZ‐mediated polyamine regulation outside of the polyamine biosynthetic pathway remains elusive. We hypothesize that OAZ sequestration results in increased polyamine uptake. To provide evidence to our hypothesis, we generated covalent OAZ‐mimetic inactivators targeting both ODC and AZIN. Using biochemical experiments, we demonstrate that these OAZ‐mimetics inhibit the activity of ODC and prevent the sequestration of OAZ by AZIN. Using Mass spectrometric experiments, we determine the covalent binding of OAZ‐mimetics to both ODC and AZIN. We employ a novel gel assay to assess the rate of covalent modification of ODC and AZIN. Finally, using computational modeling, we determine the mode of binding of OAZ‐mimetics to both ODC and AZIN to provide insights into inhibition mechanisms.
Use of DNA Barcoding to Identify Species of Raw Ready‐To‐Eat Seafood Products Sold in Orange County, CA
Authors: Tabb Amanda , Kitch Courtney , Marquis Grace , Hellberg Rosalee
Published: May 2022

DNA barcoding is widely used to identify commercial fish species sold in the U.S and internationally. This method is based on DNA sequencing of a standardized region of DNA from a sample. Sequences can then be analyzed against the Barcode of Life Data system (BOLD) for species identification. Additional analysis of samples by a method called DNA mini‐barcoding can be used for samples that cannot be correctly identified to the species level by the traditional DNA barcoding method. While relatively high levels of seafood mislabeling have been reported for sushi dishes, few studies have investigated mislabeling of other raw, ready‐to‐eat (RTE) seafood such as poke and ceviche. The objective of this study was to evaluate the presence of species mislabeling in raw, RTE seafood products in Orange County, CA. DNA barcoding and mini‐barcoding were used to investigate 70 samples of sushi, ceviche, and poke collected from various locations in Orange County. All samples underwent DNA barcoding, followed by species identification using the BOLD database. Six samples underwent further analysis with mini‐barcoding. The FDA seafood list was used to identify common and acceptable market names. The results showed that 13 samples were substituted on the basis of species, and 20 samples did not have an acceptable market name. The overall mislabeling rate for all samples combined was 52.3% (33/63). These results indicate a need for greater scrutiny of labeling practices for raw, RTE seafood sold in Orange County, California.
Auto‐inhibitory interactions of Sec7, master regulator of the Golgi complex
Authors: Brownfield Bryce A. , Fromme J. C.
Published: May 2022

In eukaryotes, protein and lipid secretory trafficking is orchestrated by the Golgi apparatus where proteins are modified and sorted in a cascade of functionally distinct compartments. The composition of these compartments is maintained primarily by vesicular traffic facilitated by Arf and Rab GTPases. These GTPases are regulated by a GTP/GDP cycle facilitated by Guanine Nucleotide Exchange factors (GEFs) and GTPase Activating Proteins (GAPs) to induce GDP/GTP exchange and GTP hydrolysis respectively. Vesicular trafficking to and from the final compartment, the trans‐Golgi network, maintains membrane flux throughout the cell with cargo recycling and secretory pathways. Arf1 and its close paralogs are responsible for virtually all vesicle biogenesis events at the Golgi complex. In budding yeast, Arf1 is activated at the trans‐Golgi network by the Arf GEF Sec7, the homolog of the human ARFGEF/BIG proteins. Previous work has determined that Sec7 is regulated by autoinhibition, positive feedback, and GTPase crosstalk, yet the mechanistic basis for these regulatory features is unknown. To understand how Sec7 is regulated, I have determined the structure of the flexible Sec7 dimer by cryoEM at 4.9A resolution. Focused refinements on individual monomer subunits improved resolution to 3.8A, sufficient for molecular model building. This structure reveals the mechanism for Sec7 autoinhibition: in the cytosol, the GEF domain interacts with the HDS domains in a manner that prevents binding to the Arf1 substrate. We also observe that the dimerization region imparts a twist between the two monomers, apparently preventing both catalytic subunits from interacting with the membrane surface simultaneously. Functional studies are currently underway to determine the physiological relevance of our structural findings.
Agomelatine Ameliorates Obesity‐Induced Kidney Injury through the Inhibition of Renal Fibrosis and Improvement of Impaired Autophagy
Authors: Promsan Sasivimon , Phengpol Nichakorn , Sutthasupha Prempree , Lungkaphin Anusorn
Published: May 2022

Introduction

Chronic kidney disease is the crucial complication that could occur in obesity. Prolong high‐fat diet consumption can stimulate reactive oxygen species (ROS) and contribute to kidney injury through various pathways. Autophagy plays an important role for maintaining intracellular homeostasis. ROS could induce impaired autophagy, subsequently led to kidney damage. Agomelatine (AGOM), a melatonin analogue, is indicated for the treatment of major depressive disorders. The antioxidant and anti‐inflammation effects have been reported in AGOM treatment in chronic mild stress‐induced depression. Previous study found that AGOM inhibited apoptosis and ER stress in the kidney under high‐fat diet condition in rats. However, the effect of AGOM supplementation on renal fibrosis and impaired autophagy in obese condition has never been elucidated.

Aims

To evaluate the renoprotective effect of AGOM on kidney function in high‐fat diet‐induced kidney injury, as well as to explore the underlying mechanisms involving autophagy signaling pathway.

Materials and methods

Male Wistar rats were received normal diet (ND) or high‐fat diet (HF) for 16 weeks. Then, the HF rats were separated into 4 subgroups including (1) HF; (2) agomelatine20 (AGOM20); (3) AGOM40, the rats were received AGOM at the dose of 20 and 40 mg/kg/day, respectively; and (4) N‐acetylcysteine (NAC), the rats were received NAC at the dose of 100 mg/kg/day by oral gavage for 4 weeks. After 4 weeks of treatment, all rats were sacrificed. Blood, urine and kidney tissues were collected for further investigations.

Results

The results demonstrated that HF rats developed kidney dysfunction as shown by the raising in serum creatinine, creatinine clearance along with the changes of glomerular structure and the elevating of positive area of α‐SMA, which is renal fibrosis indicator. These alterations were reversed by AGOM and NAC treatment. Moreover, HF rats showed renal oxidative stress as indicated by the increasing in PKCα and NOX4 expression and decreasing in antioxidant enzyme, GCLC. AGOM and NAC administrations reduced oxidative stress and restored antioxidant enzyme. Furthermore, high‐fat diet consumption markedly reduced AMPK, along with the elevation of mTOR expression. Additionally, we found that HF rats aggravated impaired autophagy which could induce oxidative stress, fibrosis and finally led to kidney cell death. AGOM administration restored autophagy process as shown by the increasing in Beclin‐1, LC3B and Atg5 protein. These results demonstrated that AGOM improved kidney injury through the regulation of oxidative stress, fibrosis and autophagy process.

Conclusions

These findings indicated that AGOM prevented kidney injury under obese condition via the inhibition of oxidative stress and fibrosis and improved impaired autophagy via regulating AMPK‐mTOR‐autophagy signaling pathways.
Development and Characterization of Fluorescent Chemical Tools to Study Human Carboxylesterase 2 (CES2)
Authors: Karns Carolyn J. , Flaig Luke H. , Singh Anchal , Gao Mingze , Beck Michael W.
Published: May 2022

The two major human carboxylesterases (CESs), CES1 and CES2 play a key role in the hydrolysis of xenobiotic esters including aspirin, methylphenidate (Ritalin), and the illicit narcotics cocaine and heroin. CES‐mediated metabolism of ester containing drugs can result in their activation or deactivation and enhance clearance. Despite the established role of CESs in drug metabolism, few techniques exist to study their activity in live cells. Fluorescent chemical tools have been developed to be able to expand the methods used to study CES activity, however, many existing fluorescent chemical tools are not sufficiently characterized to be able to confidently study CES activity in live cells. This is particularly true for CES2 where substrate specificity can overlap with other cellular esterases. To address this issue, we report the design and synthesis of new fluorescent chemical tools to study CES2. We subsequently evaluated their specificity for CES2 over CES1 in vitro and in live cells using small molecule inhibitors and shRNA. Overall, our studies resulted in a full characterization of fluorescent chemical tools for studying human CESs and will lead to improved methods for studying the role of CESs in human health and drug metabolism.
Optimizing Co‐Expression of Human Circadian Protein Complex CLOCK/BMAL1
Authors: Adame Sophia , Sanchez Sebastian , Lopez Obed , Madariaga Alberto , Moreno Brenda , Xian Yuejiao , Xiao Chuan
Published: May 2022

Circadian rhythm is the internal clock in the body that cycles in approximately 24 hours. It controls many physiological changes such as the sleep‐wake cycle and the body‐temperature fluctuation. In humans, every single cell has its own circadian rhythm that is synchronized by a master clock in the brain. At the molecular level, circadian rhythms of individual cells are controlled by the transcriptional‐translational feedback loop (TTFL). This loop is initialized by the heterodimerization of circadian locomotor outputs cycle kaput (CLOCK) and brain muscle ARNT‐like 1 (BMAL1). The long‐term goal of the project is to study the structure and the function of the full‐length human CLOCK/BMAL1 complex. Human CLOCK and BMAL1 genes have been cloned into a bacterial expression system. In order to reach the long‐term goal, it is necessary to optimize the condition of expression and purification of the CLOCK/BMAL1 complex to a high quality and homogeneity. In this study, concentration of the inducer, induction time, cell density, and lysis methods have been optimized through multiple series of expression trials. Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS‐PAGE) and Western Blotting have been used to assess the quantity and quality of CLOCK/BMAL1 expression. Protein expressed using the optimized conditions has been subjected to purification experiments. Expression and purification of the CLOCK/BMAL1 proteins will build a solid foundation for the structural and functional studies, which will deepen our understanding of the molecular mechanism of circadian rhythm that will facilitate the design of therapeutic reagents to treat circadian related diseases such as sleep‐wake disorder and jet lags.
Hypoglycemia: Control of counter‐regulatory hormone secretion by the brain
Authors: Verberne Anthony J.
Published: May 2022

Hypoglycaemia is a common side‐effect of the treatment of type 1 diabetes with insulin. The Diabetes Control and Complications Trial indicated that, while intensive insulin therapy reduced the incidence and severity of diabetic complications such as retinopathy, neuropathy and nephropathy, the incidence of severe hypoglycaemia was increased 2‐3 fold. In non‐diabetic individuals, hypoglycaemia triggers the glucose counter‐regulatory response. This response consists of inhibition of insulin secretion and activation of glucagon secretion. Prolonged hypoglycaemia also evokes adrenaline secretion and less importantly, growth hormone and cortisol secretion. However, in individuals that have type 1 diabetes, the glucagon response to hypoglycaemia is lost. In these patients, the pancreatic α‐cells continue to synthesise glucagon since glucagon secretagogues such as arginine produce a robust glucagon secretory response. Our research has examined the hypothalamic and medullary neurocircuits that mediate adrenaline and glucagon secretion under conditions of neuroglucoprivation or hypoglycaemia. These studies have shown that the response to neuroglucoprivation involves specialised sympathetic premotor neurons in the rostral ventrolateral medulla (RVLM) that mediate adrenaline secretion. These neurons are readily distinguishable from sympathetic vasomotorneurons also located in the RVLM. In addition, we have described the properties of perifornical hypothalamic (PeH) orexinergic (Ox) neurons that are activated by insulin‐induced hypoglycaemia (IHH) or neuroglucoprivation. PeH neurons project to many different targets including the RVLM and the dorsal motor nucleus of the vagus (DMV). Ox2‐R in the RVLM mediate the adrenaline secretory response. PeH neurons also mediate the glucagon response to IHH since the release of glucagon in response to IHH is diminished in Ataxin3 rats which have an age‐related loss of PeH orexin neurons.
Investigations into Multiple Acyl‐CoA Assimilation Pathways in a Single Organism
Authors: Rost‐Nasshan Aerin , Miller Stephanie , Than Naing Yadanar , Carter Michael
Published: May 2022

By 2050 there could be around 12 billion tons of plastic in landfills and the natural environment, which could be avoided by using a more environmentally friendly alternative: biologically produced plastic, or bioplastic. Polyhydroxybutyrate (PHB) is a bioplastic whose precursor is an intermediate in the ethylmalonyl‐CoA pathway (EMCP), a pathway for acetyl‐CoA assimilation in bacteria. The genes that encode the enzymes for two different acetyl‐CoA assimilation pathways, the glyoxylate bypass (GB) and the EMCP, have been observed in two PHB‐producing bacteria: Rhodobacter capsulatus and Paracoccus denitrificans. It was previously observed that P. denitrificans asynchronously uses both pathways when grown with acetate. We produced R. capsulatus fusion strains by fusing the gene for a fluorescent protein, mCherry, to the gene of a key enzyme of the EMCP or GB and observed the fluorescence during growth with acetate. Tracking fluorescence allowed us to track the accumulation of each key enzyme, which indirectly indicates the activity of the pathway. We observed that unlike P. denitrificans, the GB in R. capsulatus is not essential to growth with acetate. Understanding the use/occurrence of the EMCP in bacteria is a step to being able to manipulate the EMCP to produce commercially relevant levels of bioplastic and related products.