bluuebunny/crossref_metadata_2025_split huggingface dataset:
Application of unknowns in organismal biology laboratories to encourage inquiry‐based learning
Authors: Guenther Merrilee , Bennett Kyle , Arriola Paul 
Published: November 2023

The study of anatomy and morphology is often mistakenly seen as an exercise in memorization. Rather, morphological study is a unique opportunity for students to build their observational skill set. Observational skills are an important complement to experimental skills and crucial to students considering clinical professions. The Elmhurst Biology Department recognized the need to change the culture of certain lab experiences in order to achieve this goal and reinforce the mission of the department.The Elmhurst College Biology Department is currently utilizing a tool in the laboratory that applies concept‐based learning to building observational skills in the context of organismal and morphological education. The use of unknowns is a standard in many experimental exercises in science labs. The introduction of specimen‐based unknowns in organismal courses has encouraged the development of critical thinking skills as well as improved the mastery of anatomical content. This approach has been applied to courses across the biology curriculum, including nursing anatomy and physiology (BIO 107/108 Human Anatomy and Physiology), general biology for majors (BIO 201 General Biology II), and upper level major biology electives (BIO 330 Comparative Chordate Anatomy, BIO 430 Advanced Human Anatomy). This method has been applied in individual laboratory sessions and has also served as the basis for a semester long dissection project in the Comparative Chordate Anatomy course.The initial impact of this strategy has been apparent in terms of student engagement. Qualitatively, it has been observed that students are utilizing the complete lab period and increasing the time spent observing each specimen. The strategy appears to encourage more proactive, inquiry‐based learning by the students. By applying this approach in introductory courses, it has the potential to impact behavior in upper level courses.Preliminary attempts to assess this laboratory method were carried out in Human Anatomy and Physiology I (BIO 107) and II (BIO 108), introductory courses aimed at nursing majors, non‐biology majors pursuing health professions, and biology majors. Experimental sections were given anatomical unknowns, while the control groups were not. To control for the impact of the amount of time spent studying on student performance, all groups were required to remain in lab for the same period of time. The use of unknowns in the experimental sections was replaced with a brief instructor led review of the material in the control sections, providing a contrast of active and passive learning approaches. On lab practical exams, the experimental group outperformed the control group throughout the semester.
Arl4A interacts with Robo1 to promote cell migration via up‐regulating Cdc42 activation
Authors: Lee Fang‐Jen Scott , Chiang Tsai‐Shin , Chen Chieh‐Hsin , Tsai Meng‐Chen , Jang Li‐Ting 
Published: November 2023

Cell migration is a highly regulated event that is initiated by protrusion of the cell membrane and actin reorganization. Robo1, a single‐pass transmembrane receptor, is crucial for neuronal guidance and cell migration via activating Cdc42 GTPase. ADP‐ribosylation factor (Arf)‐like 4A (Arl4A), one of Arf small GTPases, functions in cell morphology, cell migration, and actin cytoskeleton remodelling; however, the molecular mechanisms for Arl4A in cell migration is not clear. Here, we report that Arl4A binding to Robo1 modulates cell migration via promoting Cdc42 activation. We found that Arl4A interacts with Robo1 in a GTP‐dependent manner and residual 1394‐1399 of Robo1 is both required and sufficient for this interaction. Arl4A‐Robo1 interaction is essential for Arl4A‐induced cell migration and Cdc42 activation. We also showed that Arl4A binding to Robo1 decreases the association of a Cdc42‐GAP srGAP1 to Robo1. Furthermore, Slit2/Robo1 binding decreases Arl4A‐Robo1 interaction in vivo. Thus, our study reveals a novel mechanism that Arl4A participates in Slit2/Robo1 signaling on modulating cell motility via up‐regulating Cdc42 activation.Support or Funding InformationNHRI‐EX106‐10601B1
Using 3D Paper Model Constructions and Wikki Stix&[trade] to Teach the Intricacies of the Pterygopalatine Fossa
Authors: Hatcher April , Naze Garrett , Hazzard Matt , MacPherson Brian 
Published: November 2023

PurposeThe objective of this study was to create various model representations of the pterygopalatine fossa. This small, deep area within the facial skeleton is difficult to conceptualize as the various openings into and out of the fossa are not clearly visible from inspection of the skull alone. The traditional method for teaching this region to the dental students at our university has been to use a two‐dimensional representation wire‐diagram that emphasizes the various fiber types that are transmitted through the fossa.MethodsUsing materials easily accessible to educators, such as paper and craft supplies, a team of anatomists, clinicians, and medical illustrators collaborated to design 3D models representing the complex region of the pterygopalatine fossa.ResultsTwo 3D models were created. Each used Wikki Stix, wax‐infused threads of various colors, to represent the sensory and autonomic fibers in the fossa. One kinesthetic model is a stylized inverted‐cone representation of the fossa that is pre‐labeled. The openings in the model were cut using a Cricut Explore One™ Machine. First‐year dental students were instructed how to arrange the Wikki‐Stix through the foramina of the fossa and how to fold the three‐dimensional cone model to incorporate these fibers. A second 3D kinesthetic model for teaching was developed alongside the simple model. The more complex 3D paper model used intervals of coronal, transverse, and sagittal sections through the skull to create the corresponding regions of communication with the pterygopalatine fossa, such as the cranial, oral, and nasal cavities and the orbit. These planes were then assembled into a free‐standing product. This model included black and white shadings of landmark structures such as the hard palate for the greater and lesser palatine nerves and the nasal cavity for the sphenopalatine (nasopalatine) nerves. This model was used to emphasize the spatial relationships of the nerves to approximate spaces within the cranium and skeleton. Dental students also crafted the Wikki‐Stix in context with the detailed 3D paper model to arrange the fibers in their target areas.ConclusionsIt is possible to produce 3D models of complex anatomical structures using basic materials. These models may be easily adopted by fellow educators as complex and expensive 3D printing tools are not required. Future research is needed to determine if these 3D models improve student outcomes compared to traditional teaching methods.
Heterogeneity Pattern of Apoptosis Mediators in Cervical Carcinoma Cells
Authors: García‐Aguiar Israel , Del Moral‐Hernández Oscar , Martínez‐Castillo Macario , Bonilla‐Moreno Raúl , Villegas‐Sepúlveda Nicolás 
Published: November 2023

Alternative splicing is one important regulatory posttranscriptional mechanism for control of gene expression. It produces several mature messenger RNAs (mRNAs) from single immature pre‐messenger RNA (pre‐mRNA) by using alternatively donor or acceptor splicing sites. Viruses use this posttranscriptional mechanism to increase their coding capacity, because they usually contain small genomes. The infection with the Human Papillomavirus (HPV), mainly with the types 16 (HPV‐16) and 18 (HPV‐18), is etiologically related to cervical cancer. The HPV transcripts are bicistronic or polycistronic and contain suboptimal splicing sites, which favor alternative splicing in its intron‐1; it is located in the E6/E7 oncogenes which are responsible for the transformation of host epithelial cells. Interestingly, alternatively spliced forms of the E6/E7 oncogenes are heterogeneously expressed in tumor cells and the mechanism producing the heterogeneity is still unknown. In our laboratory, it has been observed that the E6/E7 heterogeneous profile might result of differential use of acceptor sites in different cellular contexts, for example, cervical carcinoma cells with differential expression of splicing factors. Cancer cells are heterogeneous and show differential expression profiles in some metabolic markers. This fact raises the question whether the E6/E7 splicing profile heterogeneity is produced by the same mechanisms which produce tumor cell heterogeneity. Besides, some proteins involved in apoptosis are also regulated by alternative splicing; this mechanism produces alternative proteins with altered subcellular localization, as well as antagonist or dominant‐negative functions, which affect the apoptotic cell signaling and produces the phenotype of apoptosis‐resistant cancer cells. Thus, our goal was to investigate the variation in the expression profile of apoptotic mediators among cervical carcinoma cell lines. In this work we investigate the expression of alternative splicing forms of proteins involved in apoptosis, by using RT‐PCR and western blot assays in four carcinoma cell lines: C33‐A (HPV‐), CaSki (HPV‐16), HeLa (HPV‐18) and SiHa (HPV‐16). Our results revealed that some of the alternative splicing forms of the apoptotic markers were expressed differentially in both, HPV negative or positive cervical carcinoma cells; the differentially expressed alternative transcripts not presented exact correlation with the level of proteins that they encode; all these results suggested that the expression heterogeneity was produced by a complex mechanism involving several steps of post‐transcriptional control.Support or Funding InformationThis work was supported in part by funds allocated to NVS by CINVESTAV and by CONACYT
Regulation of MAP kinase signaling by c‐Myc mediates the apoptotic response to vinca alkaloids
Authors: Gristock Rachel A , Wales Christina TK , Jacobs Aaron T 
Published: November 2023

c‐Myc is a regulator of gene expression with dichotomous roles in cancer growth and viability. It is an oncogene that promotes cellular proliferation and is often over‐expressed in cancer. But c‐myc also sensitizes cancer cells to the actions of certain chemotherapeutics, including doxorubicin, cisplatin, and vinca alkaloids, among others, and is a potentially useful marker for predicting clinical responses to drug therapy. Our goal was to investigate the significance of c‐myc in mediating the apoptotic response to vinca alkaloids, and to determine the signaling pathways that relate c‐myc expression to the regulation of apoptosis. Experiments were performed in the colorectal cancer cell lines RKO and HCT‐116. Cells were transfected either with a negative control or c‐myc siRNA, then treated with vinca alkaloids (vinblastine, vincristine). Treatment with vinca alkaloids induced apoptosis in the range of 5–10 nM, as evidenced by caspase 3, 9 and PARP cleavage, and by fluorogenic assay of caspase‐3/7 activity. Vinca alkaloid treatment also caused robust JNK‐1/2 phosphorylation, which has been previously reported to promote apoptosis to these agents. Our data show that c‐myc is critical for JNK‐1/2 activation. Silencing c‐myc inhibits the phosphorylation of JNK‐1/2 on Thr183/Tyr185 in vinca alkaloid‐treated cells, and causes a corresponding reduction in drug‐induced apoptosis. The importance of JNK‐1/2 signaling was confirmed with the biochemical inhibitor SP600125, which attenuated apoptosis and increased the IC50 of vinca alkaloids by more than 20‐fold. Conversely to its effects on JNK‐1/2, silencing c‐myc enhanced the phosphorylation of ERK‐1/2, which promotes cell survival and resistance to apoptosis. Accordingly, inhibition of ERK‐1/2 with U0126 lowered the IC50 and restored apoptosis in c‐myc silenced cells. We therefore propose a model in which c‐myc facilitates JNK‐1/2 activation and suppresses ERK‐1/2, and demonstrate that when c‐myc expression is reduced, resistance to vinca alkaloid‐initiated apoptosis is acquired.Support or Funding InformationGrant support: NIH Grant P20GM103466
Analysis of the Expression of E6^E7, a Short Splicing Variant of HPV‐16‐E6 in the HPV Negative Cervical Carcinoma Cells C33‐A
Authors: Vaisman Carolina Eugenia , Bonilla‐Moreno Raúl , Villegas‐Sepúlveda Nicolás 
Published: November 2023

Human Papillomavirus (HPV) type 16 (HPV‐16) is the most important of the high risk HPV (hrHPVs) implicated in cervical cancer development. It is found in more than 60% of the cervical tumors. The E6/E7 oncogenes are responsible for the transformation of the epithelial cells; they targeted multiple proteins in the host cells, including p53 and other apoptosis mediators. The E6/E7 transcripts of the hrHPVs are first produced as bicistronic or polycistronic messages and then alternatively spliced. The alternative E6/E7 transcripts are expressed heterogeneously in the tumor cells and the impact of this differential splicing profile is not yet deeply investigated. Four transcripts are produced from the unspliced HPV16 E6/E7 pre‐mRNA: the unspliced E6/E7 RNA itself, which is the only source of the E6 oncoprotein, the E6*I/E7 and E6*II/E bicistronic transcripts, encoding two short forms of E6 transcript (E6*I and E6*II, respectively) and the E7 oncoprotein, lastly, a minor monocistronic transcript, namely E6^E7 which is formed by an in‐frame fusion of the 5′ end terminus of E6 plus the 3′ terminus of the E7; it potentially produces a hybrid protein from both the E6 and E7 oncoproteins, such fused protein could affect target of both the E6 and E7 oncoproteins. The E6^E7 transcript is also heterogeneously expressed in tumor cells but its function is poorly understood.We aim to understood the function of the HPV16 E6^E7 protein and its role in tumor cell behavior.We transfected stably HPV‐negative C33‐A cell line with a construct containing the monocistronic E6^E7 cDNA fused to the green fluorescent protein (GFP). Several stably transfected clones that expressed the fused protein differentially were selected, by using cell sorting. Proteins extracts were obtained from these clones and used for western blot analysis to investigated apoptosis effectors and members of the inhibitor apoptosis proteins (IAPs).The results showed that the transfected clones expressing different levels of the E6^E7 fused protein displayed similar levels of the IAP proteins analyzed so far. These results suggest that there is no relation between the expression level of GFP reporter gen in E6^E7 transfected C33A cells and IAPs. Conversely, the expression of the constructs containing the E6, E6*1 or E6*II cDNAs fused to the GFP protein produced an altered expression of the same IAP proteins.Support or Funding InformationCONACYT, CINVESTAV
BET Inhibition Suppresses Gastric Cancer Cell Proliferation by Inducing Cell Senescence
Authors: Dong Xingchen 
Published: November 2023

The bromodomain and extra‐C terminal domain (BET) proteins are important in mediating cell cycle control, inflammatory cytokine production and developmental programs. BET proteins bind to acetylated histone or non‐histone proteins and serve as transcriptional co‐regulators of gene expression. Small molecules targeting bromodomains of BET proteins, BET inhibitors (BETi), possess strong anti‐tumor activities and have emerged as potential therapeutics in cancer and inflammatory diseases. However, BETi target all the BET family proteins, including Brd2, Brd3, Brd4 and Brdt. The contribution of each BET protein and the detailed mechanism of BETi‐mediated inhibition of cancer cell proliferation remain to be determined.In this study, we demonstrate that BETi JQ1 suppresses the tumorigenicity of gastric cancer cells by targeting Brd4‐mediated microRNA expression. JQ1 inhibits gastric cancer cell proliferation, migration, and invasion by inducing cell senescence. Depletion of Brd4 but not other BET proteins recapitulates JQ1‐induced cell senescence. Furthermore, we demonstrate that knockdown of Brd4 or treatment of cells with JQ1 increase p21 level through down‐regulation of miR‐106b‐5p. Overexpression of miR‐106b‐5p prevents JQ1‐induced p21 expression and cell senescence, whereas miR‐106b‐5p inhibition up‐regulates p21 and induces cell senescence. Finally, we show that JQ1 abolishes the binding of Brd4 to the regulatory element of miR‐106b‐5p to inhibit the expression miR‐106b‐5p.Our results identify a novel function of Brd4 in cancer cell proliferation by modulating cell senescence via miR‐106b‐5p/p21 axis and provide new insights into using BETi as a potential anti‐cancer drug.
Targeting potassium channels in cancer: A novel strategy for therapeutic intervention
Authors: Gentile Saverio 
Published: November 2023

Breast cancer (BC) is the second leading cause of cancer‐related death in woman worldwide and accounts for more than 40.000 lives every year in the USA due to disease progression. Although the considerable success rate achieved in treating primary tumors, treatment of metastatic BC (mBC) have been severely hindered by the difficulty in targeting proteins governing important steps of the metastatic cascade, limited drug availability, side effects and drug resistance in BC patients. Therefore, finding novel therapeutic agents targeting specific molecular pathways involved in mBC are urgently needed.During our investigation to identify new and critical targets in BC, we found a novel role for the potassium ion channel Kv11.1 in BC cells and xenograft tumors. Our preliminary experiments suggest that stimulation of Kv11.1 activity with the small molecule NS1643 induces a strong inhibition of tumor growth and metastasis of BC. However, NS1643 did not produce any effects in non‐transformed breast cells and was well tolerate in animal suggesting that NS1643 molecule could be considered efficacious and safe anticancer drug.Failing of anticancer therapies is mostly due to our limited understanding of biochemical signaling in cancer cells. In this work we describe the biochemical signaling linking activation of Kv11.1 potassium channel to the tumor suppressor mechanism senescence which include a unique calcium‐dependent and p53‐independent upregulation of tumor‐suppressor p21 and novel degradation mechanism of tumor markers such as cyclin E2.Interestingly, investigating on the possible mechanism linking Kv11.1 stimulation to inhibition of metastasis revealed that Kv11.1 agonists strongly inhibit cancer cell motility. Our experiments revealed that NS1643 strongly down‐regulated epithelial to mesenchymal transition (EMT). Although it has been well established that EMT is a process by which cells gain migratory properties and strongly contributes to metastasis, still very little is known about the biomolecular mechanism that regulates EMT.We describe for the first time the mechanism linking activity of Kv11.1 potassium channel to suppression of EMT markers and increased cell‐cell contact.Current therapeutic approaches against metastatic BC can aim only to ameliorate some of the symptoms and their clinical efficacy is strongly limited by the severity of their side effects. We think that this study offers an unprecedented opportunity to design a novel therapeutic approach against mBC in which Kv11.1 activators can be used to suppress cancer cell growth and dissemination.
Inhibition of Human Prostate Cancer Growth by Mesenchymal Stem Cells Delivering MiR‐16
Authors: Jones Eyone , Mazirka Pavel , McNurlan Margaret A , Brink Peter , Caso Giuseppe 
Published: November 2023

We have previously shown that miR‐16 has suppressive effects on the growth of human prostate cancer cells (PC3). However, one of the limitation for the use of inhibitory microRNAs (miRs) or small interfering RNAs (siRNA) for cancer therapy is effective delivery to cancer cells in vivo. The aim of this study was to evaluate the use of human mesenchymal stem cells (hMSC) as a potential in vivo delivery system for miRs to cancer cells.Human xenograft tumors were established by s.c. injection of 1 × 106 human prostate cancer cells (PC3) in immunocompromised nude mice. When tumors reached a measureable size, mice were randomized to receive hMSC previously transfected with miR‐16 (n=10, Treated) or saline (n=10, Control). 1 × 106 hMSC or saline were injected intratumorally. Tumor growth was assessed over a period of 3 weeks.Tumors in the group treated with hMSC transfected with miR‐16 showed a slower growth rate. After 3 weeks the average tumor volume was 40% lower in the treated group compared to controls (1.2 ± 0.22 vs. 2.04 ± 0.46 cm3). The increment in tumor size over baseline (pre‐treatment) values was 5.0 fold in the treated and 8.4 fold in the control group (P<0.05). Tumor weight was also significantly lower in treated animals (0.82 ± 0.15 vs. 1.2 ± 0.16 g, P < 0.05).In summary, treatment of established xenograft tumors derived from human prostate cancer cells (PC3) with hMSC previously transfected with miR‐16 significantly slowed tumor growth. The results confirmed that miR‐16 can suppress tumor growth and indicate that hMSC could represent a suitable system for in vivo delivery.Support or Funding InformationSupported by the Stony Brook Foundation TRO‐Catacosinos Cancer Translational Researcher Award, and by the Dept. of Surgery, Stony Brook University Medical Center (Small Grant Program)
Subpopulations of Dopamine Transporters Show Enrichment in Phosphorylation or Palmitoylation
Authors: Storandt Michael H , Hovde Moriah J , Stanislowski Daniel J , Vaughan Roxanne A , Foster James D 
Published: November 2023

The dopamine transporter (DAT) is a key regulator of duration and intensity of dopaminergic signaling through its uptake of dopamine from the extraneuronal space, and DAT dysregulation has been hypothesized to play a role in dopaminergic diseases such as autism, attention deficit hyperactivity disorder, and drug addiction. DAT consists of 12 transmembrane spanning domains that form the substrate permeation pathway, and cytoplasmic N‐ and C‐termini that contain sites for regulatory post‐translational modifications. Phosphorylation of the N‐terminus occurs on Ser7 and Thr53, which leads to decreased transport Vmax, and S‐palmitoylation occurs on Cys580 at the membrane‐cytoplasm interface of the most C‐terminal transmembrane helix, which leads to increased transport Vmax. We previously showed that palmitoylation and phosphorylation occur in a reciprocal manner, with high phosphorylation promoting reduced palmitoylation and visa versa, consistent with their opposing functional outcomes, but it has not been established if these modifications can coexist on a single molecule of DAT or if they are mutually exclusive. To address this issue, we isolated palmitoylated DAT by acyl‐biotin exchange (ABE) and neutravidin affinity chromatography, followed by phospho‐specific immunodetection of Thr53 phosphorylation. Our results indicate that only a small percentage of total DATs are simultaneously phosphorylated and palmitoylated, indicating that the modifications are predominantly mutually exclusive. We also found that S7A DATs display increased palmitoylation, consistent with previous findings, but also show a trend towards increased Thr53 phosphorylation, indicating complex interplay between all three of these modifications. These results support the presence of communication between subregions of the N‐terminus as well as between N‐ and C‐termini, and highlight the mechanistic integration of these post‐translational modifications in the establishment of dopamine reuptake capacity.Support or Funding InformationNational Institutes of Health grants DA 031991 (JDF), DA13147 and 5P20‐104360 (RAV), P30‐GM103329 (UND), P20‐GM12345 (UND)