bluuebunny/crossref_metadata_2025_split huggingface dataset:
Using 3D‐Printing to Evaluate Trabecular Bone Mechanical Properties
Authors: Barak Meir M , Black Arielle M 
Published: November 2023

IntroductionTrabecular bone structure is essential to the normal mechanical behavior of whole bones. Studies have shown that osteoporosis negatively affects trabecular bone structure, mostly by reducing trabecular number and decreasing the thickness of the remaining trabeculae. The end effect of these phenomena is an increase in fracture risk. A major difficulty in assessing and quantifying the effect of trabecular structural deterioration on the mechanical behavior of whole bones is that no two trabecular structures are identical. Thus, when we compare a group of healthy vs. osteoporotic samples we only get the general trend. 3D‐printing is a new technology that can assist in overcoming this issue. Here we present a preliminary study that compares a healthy 3D‐printed trabecular bone model with the same model after osteoporosis was simulated (i.e. trabecular number and thickness were reduced). Since the osteoporotic model is derived from the healthy one, it is possible for the first time to directly estimate (percentage wise) the decrease of tissue stiffness and strength as a result of trabecular bone structural deterioration.MethodsA cubical volume of interest (4.5×4.5×4.5 mm) was cropped and segmented from a micro‐CT scan of a chimpanzee's metacarpal head using Amira 6.0. To simulate the onset of osteoporosis (loss of about 10% of bone mass), a second model was created by raising the gray‐scale threshold during segmentation. Next, the “healthy” and “osteoporotic” models were printed in scale (1:1 to real life size) 30 times each, using a 3D‐printer (ProJet 1200). Finally, all samples were tested in compression until failure (Instron 5942) and trabecular bone tissue strength and stiffness was calculated.Results and discussionOur results demonstrate that the “osteoporotic” trabecular structure was significantly less strong and stiff than the original intact (healthy) structure (P<0.01). Structural strength decreased by 47%, from an average of 4.13±2.04 MPa to 2.19±0.77 MPa, while structural stiffness decreased by 35% from an average of 71.82±20.30 MPa to 45.36±12.73 MPa. Despite the fact that these 3D models are made of printing resin and not bone, they enable us to compare the exact same trabecular structure before and after the onset of trabecular tissue structural damage. Our results demonstrate that a relatively small decrease in bone volume (about 10% of bone mass) leads to a dramatic decrease in structural strength (47%) and stiffness (35%).ConclusionsThis study demonstrates that 3D‐printing is a valuable tool for estimating the mechanical properties of trabecular structures. In the future, 3D‐printing may help us to attain better personal fracture risk assessments and consequently improve life quality for people suffering from Osteopenia and osteoporosis.Support or Funding InformationThe study was supported by the Winthrop University Research Council Grant (SC15014).
Urinary proteome changes were detected earlier than serum biochemical parameters and histopathology changes in a rat thioacetamide‐induced hepatic fibrosis model
Authors: Zhang Fanshuang , Ni Yanying , Yuan Yuan , Gao Youhe 
Published: November 2023

Whereas plasma remain homeostatic, urine tends to accumulate and reflect early changes of the body. This property potentially makes urine a better biomarker source for diseases research. Liver fibrosis which results from depositing extracellular matrix (ECM) components is still a reversible pathological condition whereas cirrhosis as the end‐stage of liver fibrosis is irreversible. Consequently, noninvasive early biomarkers are desperately needed. In this study, thioacetamide (TAA) peritoneo‐injection rat model was used to identify differential urinary proteins in liver fibrosis process by using tandem mass tagging and two‐dimensional liquid chromatography tandem mass spectrometry. A total of 766 urinary proteins were identified, 143 and 118 of which were significantly changed in TAA 1‐week and 3‐week groups respectively, compared with the control group. Multiple reaction monitoring (MRM) targeted proteomics was used for further verification of differentially proteins. A total of 40 urinary proteins were statistically significantly changed (p<0.05) and had fold change more than 2. Those urinary protein changes were earlier than serum biochemical parameters aspartate aminotransferase (AST), alanine aminotransferase (ALT), albumin, total protein and histopathology, which have potential as early biomarkers of liver fibrosis.Support or Funding InformationThis work was supported by the National Key Research and Development Program of China (grant number 2016 YFC 1306300); the National Basic Research Program of China (grant number 2013CB530805); and funds from Beijing Normal University (grant number11100704, 10300‐310421102).
Absolute Quantification of RXRG isoforms
Authors: Skotarczak Teagan , Karnath Erin , Monin Lydia , Shimanovich Yana , Bistulfi Gaia 
Published: November 2023

Many genes are transcribed as multiple variants known as isoforms. Isoforms are transcripts from the same locus with either a different transcription start site or alternative splicing. Isoforms might be characterized by different functions and/or stability. Different isoforms of the same gene can vary in length from a few bases to several kilobases. In some cases, there is no region of the transcript specific to only one isoform, making quantification problematic. We compared the isoforms' sequences using bioinformatic tools freely available online. We utilized a one‐step real‐time RT‐PCR protocol, using primers specific to one or more isoforms, which included a T7 promoter and terminator sequences. After confirming amplicon identity by DNA sequencing, we built RNA standard curves. Analysis of the data shows acceptable specificity and efficiency for RXRG3 primers as well as primers picking up all three isoforms. We are currently in the process of assessing primers for RXRG1 and RXRG3.Support or Funding InformationThis work was supported by funding from D'Youville College, Buffalo NY
In Vitro Activation of Purified Aedes Aegypti Mosquito Early Trypsin (AaET) Wild‐type in Comparison to the Inactive AaET Mutant (Active Site Serine to Alanine)
Authors: Lucero Rachael M. , Rascon Alberto A. 
Published: November 2023

The female Aedes aegypti mosquito requires specific midgut proteases to degrade blood meals in a biphasic manner for completion of the gonotrophic cycle. Upon acquisition of multiple blood meals, the female anthrophophilic Aedes aegypti mosquito may become infected with several viruses, such as Dengue, Yellow fever, Chikugunya, and Zika viruses. Once infected, the mosquito can transmit these pathogens to its human hosts, making the mosquito an efficient biological vector. Current vector control strategies have not shown to be efficient, therefore studying and potentially inhibiting the required proteases for blood meal digestion could provide a new vector control strategy. This study is focusing on Aedes aegypti Early Trypsin (AaET), an early phase, trypsin‐like serine protease. This enzyme is one of four most abundant midgut proteases involved in blood meal digestion. The specific activity role of AaET wild‐type (WT) in blood meal digestion has yet to be fully elucidated. The enzyme has been solubly expressed using bacteria as a host, which has led to auto‐catalytic activity. Despite this, zymogen AaET (WT) has been successfully purified and activated using a specific buffer exchange approach that removes dithiothreitol (a reducing reagent that was added in order to inhibit auto degradation during the purification process). Proteolytic activity of zymogen and the active form of AaET (WT) were determined using a chromogenic trypsin substrate, BApNA, to compare to published results. To determine if AaET (WT) is truly autocatalytic, an AaET Mutant (MU) was constructed by mutating the Serine residue in the active site to an Alanine, and was subsequently purified to compare to the wild type form.Support or Funding InformationSC3 Research Grant from NIH
Funcionalized Gold Nanoparticles as Potential New Treatment for Acute Myeloid Leukemia
Authors: Gaiser Ann‐Kathrin , Hafner Susanne , Schmiech Michael , Syrovets Tatiana , Simmet Thomas 
Published: November 2023

Surface functionalization of nanoparticles is often used to target specific cell types or cellular organelles. We have previously found that positively charged nanoparticles manufactured from biologically inert polystyrene are highly cytotoxic towards acute myeloid leukemia cell lines while primary macrophages of healthy donors are resistant.Acute myeloid leukemia (AML) is a malignancy of the myeloid lineage of the hematopoietic system and, to date, the gold standard treatment combines two chemotherapeutic drugs ‐ cytarabine and daunorubicin. The prognosis and treatment outcome of AML strongly depends on the age of the patients. While patients younger than 65 years show a 5‐year overall survival rate of around 50%, older patients show a very poor 5‐year survival of only 4%. Hence, there is still a need for new agents with higher specificity towards AML cells and lower general cytotoxicity.In the current study we produced and characterized new amino‐functionalized gold nanoparticles and investigated their potential to target specifically leukemia cells, in vitro and in vivo.The amino‐functionalized gold nanoparticles (AuNP+) exhibit a positive surface charge, and, like the polystyrene particles, proved to be extremely cytotoxic towards AML cells.The type of cell death was identified to be apoptosis. Further analysis of the underlying mechanism revealed an early loss of mitochondrial functions, including inhibition of mitochondrial respiration and mitochondrial membrane permeabilization. Consequently, this led to release of cytochrome C into the cytosol and caspase activation.In contrast to positively charged AuNP+, non‐functionalized (AuNP) or carboxy‐functionalized gold nanoparticles with a negative surface charge (AuNP‐) did not show any cytotoxic effect. This differential toxicity of the gold nanoparticles might be due to higher uptake of positively charged AuNP+ particles by AML cells, as detected by transmission electron microscopy.The positively charged particles also induced cell death in primary human leukemia cells and reduced their colony‐forming potential, while healthy hematopoietic cells were not affected by treatment with AuNP+.Overall, we have found so far that our positively charged gold nanoparticles induce apoptosis and exhibit selective cytotoxicity towards AML cells. Further we aim to understand the cause of higher sensitivity of AML cells and to investigate the efficacy of AML treatment by AuNP+ particles in a mouse model engrafted with patient‐derived primary AML cells.Support or Funding InformationVolkswagenStiftung
Values and Perceptions of the Traditional Gross Anatomy Courses at Tufts University School of Medicine
Authors: Jarrah Andrew 
Published: November 2023

Human Gross Anatomy has customarily been taught as a didactic course supplemented with practical laboratory instruction. However, given strong directives from the Liaison Committee on Medical Education and other governing bodies, along with advances in educational technology tools, many medical curricula have forgone this format in favor of group‐based learning modalities such as the flipped classroom and Team‐based problem solving. While studies suggest that collaborative learning and technology assisted teaching may improve student performance, other data suggest that didactic formats are actually the preferred learning mode. Tufts University has maintained a traditional lecture and gross lab format with nearly 140 hours of instruction spread over five months of the first year curriculum. The longitudinal nature of the course has allowed for a paced, deep learning of Anatomy in stark contrast with shorter courses which emphasize broad or conceptual learnings. Our preliminary report, which draws on polling results and conversations with the first year class suggests that Gross Anatomy, and the complementing Head & Neck Anatomy course, are one of the most valued components of the curricula at Tufts, not only for their subject matter, but for the breadth and depth as well. Students found that the hands‐on nature of the lab work not only allowed for detailed learning of anatomical complexities, but it also vested a strong respect for the sensitivity of the profession. The pre‐lab lectures were deemed an appropriate primer for all the dissections throughout the course. The current format was found to allow for a mastery of the material due to the comprehensive nature of the didactics along with practical experience in the gross lab. The continued success of this course signals a place for traditional Anatomy instruction in a landscape which continues to evolve towards alternative learning approaches. While controversies still exist, the traditional didactic has proven its ability to deliver deep‐level learning in a format led by experts. Further studies will be needed to show whether technology or peer based learning tools are able to deliver similar results.Support or Funding InformationThis work was made possible by the American Academy of Anatomists and Tufts University School of Medicine.
Effects of Charged Resveratrol Derivatives on Ca2+ Homeostasis in Human Cancer Cells
Authors: Peterson Joshua Allen , Hastings Jordan Parker , Kenealey Jason Donald 
Published: November 2023

Resveratrol (RES), a naturally occurring polyphenol found in grapes, peanuts, and berries, selectively facilitates apoptosis in cancer cells. The apoptotic activity of RES in tumor cells is, in part, due to a large, sustained increase in cytosolic calcium ion concentration ([Ca2+]i). Via live cell fluorescence imaging, RES was previously shown to inhibit the plasma membrane Ca2+‐ATPase (PMCA) as a potential part of the apoptotic mechanism in MDA‐MB‐231 human breast cancer cells. The RES‐mediated modulation of [Ca2+]i in cancer cells may also impact other pumps and channels associated with [Ca2+]i homeostasis. The mechanism of action by which RES induces PMCA inhibition is currently unknown. We recently synthesized several charged RES derivatives that allowed us to more thoroughly study the interaction between RES, PMCA and calcium signaling. The membrane‐impermeable, charged RES derivatives are unlikely to be metabolized into a membrane‐permeable RES derivative due to the lack of membrane‐bound esterases in the cancer cell line that we studied. Utilizing live cell microscopy, we were able to determine the extracellular effects of charged RES derivatives on calcium signaling and PMCA inhibition in cancer cells.Support or Funding InformationThis research was funded by a BYU Life Sciences Startup Grant and a BYU Office of Research and Creative Activities (ORCA) Undergraduate Research Grant.
Use of Novel Drug Carrier System to Selectively Deliver Rolipram to the Liver for the Treatment of Alcoholic Liver Disease
Authors: Rodriguez‐Alvarez Walter , Zhang JingWen , Perez‐Abadia Gustavo , McClain Craig , Barve Shirish , Maldonado Claudio , Gobejishvili Leila 
Published: November 2023

Work done with phosphodiesterase (PDE) inhibitors has demonstrated their beneficial effects in experimental liver injury. Our group has shown that upregulation of hepatic PDE4 plays a pathogenic role in the development of alcoholic hepatic steatosis as well as cholestatic liver injury in animal models. Due to the potent anti‐inflammatory action of PDE4 inhibitors, two PDE4‐specific inhibitors have been recently approved by FDA for COPD and psoriatic arthritis, however both drugs have CNS associated side effects. Based on our studies demonstrating that hepatic PDE4 upregulation contributes to liver injury, we aimed to develop a liposomal‐based drug delivery system to target the PDE4 inhibitor (rolipram) to the liver, in order to achieve effective therapeutic levels while limiting its access to the CNS and preventing side effects. We demonstrated that our specially formulated and sized fusogenic lipid vesicles (FLVs) begin to selectively accumulate in the liver 10 min after administration. Moreover, when we used FLVs as carrier for rolipram (FLVs‐rolipram), endotoxin‐inducible PDE4 activity was inhibited in the liver without any effect on brain PDE4 activity. Since endotoxemia and increased PDE4 expression plays a critical role in the development of experimental alcoholic liver disease, we then tested the effect of FLVs‐rolipram in a mouse model of alcoholic hepatitis. C57Bl/6 mice were fed the Lieber‐DeCarli liquid diet containing 5% ethanol for 10 days. On day 11, they were gavaged (binged) with 31.5% (vol/vol) ethanol, which is similar to the binge drinking pattern in many alcoholic hepatitis patients. FLVs‐rolipram was administered 12 h before alcohol gavage. Control mice, on an isocaloric liquid control diet, were gavaged with maltose dextrin instead of alcohol. The mice were sacrificed 6 h after alcohol binge. Assays for liver enzymes, liver caspase 3, TUNEL, chloroacetate esterase (CAE), Oil red O staining were performed to document liver injury, steatosis and inflammation. Our results show that FLVs‐rolipram treatment markedly attenuated alcohol induced lipid accumulation in the liver and decreased plasma endotoxin levels. Moreover, hepatic apoptosis and injury was significantly lower in mice treated with FLVs‐rolipram as indicated by decreased hepatic caspase‐3 activity, TUNEL, CAE staining and AST levels. These results demonstrate that the targeted delivery of rolipram to the liver by using our novel FLV drug delivery system, achieves effective therapeutic levels in the liver, while limiting its access to the CNS and preventing side effects.Support or Funding InformationThe work was supported by following grants from NIH: R43AA021331, R44AA021331, R21AA022189, U01AA021901 and P50AA024337
Targeting the transcriptional kinases CDK12 and CDK13 in breast and ovarian cancer
Authors: Hamman Kristin , Bradley Michael , Marineau Jason , Choi Yoon , Malojcic Goran , Orlando David , Ren Yixuan , Ke Nan , Hu Shanhu , Olson Eric , Fritz Christian , Roberts Christopher 
Published: November 2023

CDK12 and CDK13 regulate expression of large transcripts requiring substantial processing to produce mature mRNA. This transcriptional regulation includes coordinated phosphorylation of specific repeats within the C‐terminal domain of RNA polymerase II and association with RNA processing factors (Chila, 2016). RNAi knockdown of CDK12 in cell culture decreases expression of DNA damage response genes, including BRCA1 and ATR, while enhancing sensitivity to DNA damaging agents (Blazek, 2011; Liang, 2015). Recently THZ531, a selective covalent inhibitor of CDK12 and CDK13, was shown to decrease expression of DNA damage response genes in cell culture (Zhang, 2016). Here we present further studies with THZ531 to guide our discovery program toward molecules suitable for clinical development and to explore mechanistic rationales for combining a CDK12/13 inhibitor with PARP inhibitors or DNA damaging agents for difficult‐to‐treat cancers such as high‐grade serous ovarian cancer and triple‐negative breast cancer.Using THZ531 as a benchmark, we developed assays capable of discriminating sub‐nM inhibitors, including quantifying time‐dependent covalent inhibition and cell‐based CDK occupancy. Since CDK7, like CDK12 and CDK13, contains a cysteine residue proximal to the kinase active site, these approaches are critical to understand covalent inhibitor selectivity. Furthermore, we performed kinome paneling studies to better understand selectivity of this scaffold in support of our ongoing efforts to optimize CDK12/13 potency and selectivity.To pharmacologically investigate the previously reported effects of CDK12 RNAi, growth inhibition of a panel of ovarian and breast cancer cell lines was assessed following treatment with THZ531 (OVA EC50 = 50–200 nM (n=6); BRCA EC50 <50 nM (n=4)). Expression profiling revealed that THZ531 treatment resulted in different sets of genes being affected than was observed following treatment with inhibitors targeting CDK7, CDK9 or BET‐bromodomain proteins. Additionally, CDK12/13, CDK7, and CDK9 inhibitors were profiled in a broad cell line panel (n>400) to reveal relationships between inhibitor sensitivity, mutation status, gene expression, and potential oncology indications that may be addressed by these different mechanisms. Finally THZ531 was synergistic with both PARP inhibitors and DNA damaging agents in ovarian and breast cancer cell lines. These data highlight cancer indications and combinations that may be particularly amenable to treatment with CDK12/13 inhibitors.While the pharmacokinetic properties of THZ531 preclude adequate target engagement in tumor tissue at tolerated doses in mouse model systems, our ongoing medicinal chemistry program is progressing to identify and optimize CDK12/13 inhibitors suitable for clinical evaluation.
The UDP‐glucuronosyltransferase (UGT) 2A1308Arg variant isoform: Its role in modulating wild‐type UGT2A1 activity and its association with lung cancer risk
Authors: Sutliff Aimee Kristina , Bushey Ryan , Ashmore Joseph H. , Lazarus Philip 
Published: November 2023

The UDP‐glucuronosyltransferase (UGT) gene family of enzymes are important in the detoxification of a variety of endogenous and exogenous compounds including hormones, bile acids, drugs and carcinogens. UGT2A1 is expressed in a variety of aerodigestive tract tissues and exhibits glucuronidation activity against polycyclic aromatic hydrocarbons (PAHs). The polymorphic UGT2A1308Arg isoform, encoded by the UGT2A1*3 allele, was previously demonstrated to exhibit no activity against a panel of tobacco carcinogens including a variety of PAHs. In the current study, the effects of UGT2A1308Arg on wild‐type UGT2A1308Gly activity and its potential role in lung cancer risk were investigated. A significant decrease (p<0.005) in UGT2A1 activity was observed against multiple PAH substrates following the co‐expression of UGT2A1308Gly and UGT2A1308Arg in the HEK293 cell line. Co‐immunoprecipitation experiments indicated that negative regulation of wild‐type UGT2A1 activity was due to protein‐protein interactions between the UGT2A1308Gly and UGT2A1308Arg isomers. In a panel of 117 human lung microsomal specimens, the glucuronidation of 3‐hydroxybenzo[a]pyrene (3‐OH‐BaP) was significantly (p<0.05) decreased in specimens from subjects with UGT2A1 (*1/*3) heterozygous alleles versus homozygous UGT2A1*1. No 3‐OH‐BaP glucuronide formation was observed in specimens from subjects with the UGT2A1 (*3/*3) genotype. In case‐control studies, the UGT2A1308Arg variant was found to be significantly associated with increased risk for non‐small cell lung carcinoma (OR=1.4; 95% CI=1.02–1.90) when comparing UGT2A1 (*1/*1) versus [(*1/*3) + (*3/*3)] subjects, a pattern that was strongest in women (OR=1.7; 95% CI=1.06–2.53). The negative modulation of wild‐type UGT2A1 activity against PAHs by the UGT2A1308Arg variant may provide a novel explanation for the variation in enzyme activity observed between individuals and may have important implications in terms of the role of the UGT2A1308Arg variant on tobacco‐related cancer risk.Support or Funding Information

National Institutes of Health, National Institutes of Environmental Health Sciences [Grant R01‐ES025460]
Health Sciences and Services Authority of Spokane, WA [Grant WSU002292]