bluuebunny/crossref_metadata_2025 huggingface dataset:
The Effectiveness of the Hybrid Teaching Method in Introductory Biology at a PUI
Authors: Resendes Karen K , Mroz Kevin , Campbell Christina
Published: June 2021

Hybrid courses combine online education (internet resources including tutorials, videos, and activities) and traditional lecture‐style classes (direct teacher student interaction) into a single course. We are studying the effectiveness of the hybrid method at a small liberal arts college with class sizes of < 24 students. We compare the hybrid and traditional methods with the same set of students, by alternating between styles weekly. Hybrid weeks include online work (quizzes on text book readings and reinforcement activities) in addition to lecture and laboratory. We applied this approach to a broadly focused non‐majors biology course geared toward education majors (Bio101) and an introductory biology course for majors that covers topics in biochemistry, cell and molecular biology (Bio201). Retention of information was measured as student improvement between a pretest and midterms or final exam, comparing retention of information by topic and by Blooms Taxonomy. In Bio101 students showed a significant difference in their ability to answer questions that required them to analyze information presented in hybrid weeks over that of non‐hybrid weeks. Preliminary results from Bio201 show no significant difference in improvement between teaching styles based on content; however, we have observed a significant improvement in analyzing and applying questions from hybrid weeks over non‐hybrid weeks.
Using web‐based media to teach first year biology students about statistical significance
Authors: Krontiris‐Litowitz Johanna
Published: June 2021

The objective of this study was to determine if web‐based media could be used to teach first year biology students statistical significance analysis and to compare this form of instruction to traditional lecture based delivery. Two methods were used to present information about statistical significance and p values, a slide presentation and 2 YouTube videos. The slide presentation was delivered by the instructor during class. The YouTube videos were embedded in a class assignment where students were instructed to analyze data, interpret results and present data in a graph format. A pre‐course assessment of student knowledge showed that 19.9% of the class knew how to use p values and recognized their importance in evaluating data. Student understanding assessed after the slide presentation showed that 23% could use significance values to correctly interpret data in a journal article and 29 % could use significance to interpret experimental data in a class quiz (p>;.05, n=60). Student understanding was assessed again after students watched the YouTube videos and an improvement observed. At this time 63.6% of the class could use significance values to correctly interpret data in a journal article (p<.05, n=60). This study shows that web‐based media can be used to teach students analytical skills such as population testing, and that in some cases this option can be more effective that traditional lecture presentations.
Minority Travel Fellows Demonstrate Successful Career Development in Long‐Term Study
Authors: Bruthers C. Brooke , Frank Martin , Matyas Marsha Lakes
Published: June 2021

Participation in professional meetings builds critical presentation skills and professional networks; both are especially important for trainees from underrepresented minority groups. Travel fellowships, such as the APS/NIDDK Minority Travel Fellows (MTF) Program, seek to address this issue. In the current study, 124 past MTFs were surveyed to assess program impacts. The survey provided information on career development (education, training positions, papers published, grants submitted, etc.), networking (continued contact with other MTFs and mentors), and perceived program impacts. Results show that the vast majority (94%) of past MTFs have continued in their physiological studies. About half (47%) of respondents were employed physiologists, 25% were postdoctoral fellows, and 17% were graduate students. Past MTFs demonstrated varying levels of career development, with success in both publishing and research funding. The vast majority spent a significant part of their time engaged in physiology or physiology‐related research. The large majority of past MTFs pointed out strong benefits received from the program. Overall results suggest that the program has long‐term positive impacts for MTFs and provides a program model for other professional societies. This program and study were sponsored by a grant from the National Institute of Digestive, Diabetes, and Kidney Disease (NIDDK R13 DK39306).
Developing an interdisciplinary science community through curriculum materials and a summer science orientation camp
Authors: Kruchten Anne E.
Published: June 2021

Faculty in the biology, chemistry, and physics departments at Linfield College are developing an interdisciplinary STEM community through two new programs. First, we have created curriculum materials that can be used to engage students across the traditional introductory science courses in scientific discussions of related topics. We will describe how we have used these materials in an interdisciplinary science seminar and how we are beginning to use them as stand alone modules in existing introductory courses. Second, we have recently concluded our first annual iFOCUS camp for incoming students. iFOCUS (Interdisciplinary First‐Year Camp for Undergraduate Sciences) engaged ten new Linfield students in collaborative research projects with Linfield faculty for a week before classes began this fall. We will report on the successes of this program, including plans for the future development of interdisciplinary science at Linfield. Funding for this program was provided by Dr. Thomas Hellie, president of Linfield College.
Benefits of using a nationally normed biochemistry examination
Authors: Linenberger Kimberly , Holme Thomas A.
Published: June 2021

The American Chemical Society (ACS) Exams Institute produces nationally normed exams for most chemistry courses, including biochemistry. The ACS Biochemistry Exam is created by a committee of educators who teach a biochemistry course. Topics typically covered on the ACS Biochemistry exam include metabolism, genetics, biomolecules, and enzymes. The 2013 Biochemistry exam is designed for use in either one‐semester biochemistry courses or two‐semester sequences. It does this by having 40 core items (for a 1‐semester course) and an additional 20 advanced items for those who use the exam in a two‐semester sequence. Discussion will focus on the benefits of using the ACS Biochemistry Exam as a form of assessment in a biochemistry course.
Teaching Circulatory Responses to Exercise Using a Classic Paper by Grimby et al.
Authors: Tanaka Hirofumi
Published: June 2021

The circulatory system must respond to the increased muscular demand for oxygen and nutrients during exercise. Here we describe our didactic approach to convey how these adjustments are achieved using the classic paper by Grimby and colleagues published in J Appl Physiol in 1966. This paper is an ideal vehicle to do so because of the comprehensive battery of tests and the integrative nature of the experiments. Following the discussion on the tight matching between metabolism (oxygen consumption) and systemic blood flow (cardiac output), factors acting to increase oxygen consumption during exercise will be illustrated using the Fick equation and the ohm's law. Because stroke volume was measured in both supine and sitting positions and heart volume was also obtained, the importance of preload or venous return on augmenting stroke volume is discussed. The students can also develop appreciation for the automated acquisition of hemodynamic measures in modern laboratory since each sample was analyzed using the time‐consuming traditional procedure (eg, Douglas bag combined with the Scholander techniques for oxygen consumption). Finally, as the data were obtained in middle‐aged and older well‐trained adults and the data on young adults previously reported are also included, declines in aerobic capacity with age and the mechanisms underlying such decline can be discussed using the Fick equation.
Regional variations in cartilage thickness of the radial head; implications for prosthesis design
Authors: Yeung Celine , Deluce Simon , Willing Ryan , Johnson Marjorie , King Graham J.W. , Athwal George S.
Published: June 2021

Background

Knowledge of potential variations in cartilage thickness is important in designing radial head implants.

Objective

To characterize the regional variations in radial head cartilage thickness.

Methods

Twenty‐seven cadaveric radii were dissected and scanned with computed tomography (CT) in neutral position. Three dimensional models were generated from CT scans and processed through a computer program by two independent observers. Cartilage thickness values were obtained at 41 predetermined landmarks located around the articular dish and side of the radial head.

Results

At the side of the radial head, cartilage in the posteromedial quadrant (0° to 90°) was significantly thicker than all other quadrants (p < 0.05). Cartilage thickness measurements within the articular dish were similar at a mean of 0.98 ± 0.13mm, but increased towards the rim of the radial head (p < 0.001). Regional variations of cartilage thickness within the rim circumference were also detected, with the thickest region located anteriorly (270°, p < 0.001) and thinnest region laterally (180°, p < 0.05).

Conclusions

Regional variations in cartilage thickness exist around the side of the radial head and within the articular dish. Such differences offer insight into the biomechanics of the elbow joint and may also affect overall radius length and dish depth, which may be clinically important for implant design.

Funding:

Departmental

Grant Funding Source: Departmental
Foraminal stenosis and anomalous venous drainage in complex craniosynostosis
Authors: Mussell Jason C , St. Hilaire Hugo , Sanchez Ruston
Published: June 2021

Craniosynostosis, the premature fusion of one or more cranial sutures, is a relatively common congenital defect, affecting ~1/2,500 live births. Craniosynostosis alters skull and brain development and may lead to facial deformities, raised intracranial pressure, cranial venous hypertension, airway obstruction, and exorbitism. Complex craniosynostosis, defined as the fusion of two or more sutures, occurs in 5 to 15% of all cases and is termed syndromic when associated with a pattern of extracranial deformations, typically involving a fibroblast growth factor receptor (FGFR) gene mutation.

Craniosynostosis requires operative intervention to remodel the skull and allow adequate brain development. It is vital during preoperative evaluation of children with multiple craniosynostosis to investigate the possibility of anomalous basicranial venous drainage. The detection of abnormally enlarged emissaries serving as major sources of intracranial venous drainage may counter indicate intervention, and it has been reported that disruption of such venous channels during surgery can lead to a fatal rise in intracranial pressure.

While venous anomalies are common in children with complex craniosynostosis, there are currently only a limited number of case reports that detail the specific routes of posterior dural sinus drainage in this population
Development of a viewing system for a whole tissue from multiple tissue blocks
Authors: Hashimoto Noriaki , Tanaka Toru , Snuderl Matija , Yagi Yukako
Published: June 2021

Viewing whole tissue allows us to recognize spatial perspective, which cannot be seen from a single glass slide and could leads to discovery of novel morphologic and growth patterns of human tumors. However, we cannot observe the whole histological image with large tissue organ. Whole Slide Imaging (WSI) can digitize the entire tissue on a glass slide. If we can use the digital images of multiple tissue blocks, it enables to view whole organ. In addition, this method can keep general versatility as conventional WSI scanners are used. Therefore, we propose a system to view a whole large tissue from multiple tissue blocks.

First, in this system, whole organ is sectioned into blocks and H&E slides are scanned by WSI scanners. Then, low‐resolution images are used to export location information (Fig. 1). In the viewer, higher magnification images of the region specified by a user are displayed on a monitor using exported information (Fig. 2).

To show the validity of the proposed system, we developed the viewer and confirmed that a whole organ can be observed efficiently. This system can makes it easier to compare to other modality images such as gross pictures of a whole organ. Furthermore, if we expand this system to sections in depth, it would allow us to compare with MRI or other medical imaging techniques. This system would be helpful for clinicopathological‐imaging correlations and better understanding of pathological processes.
Improved color visualization of histological structures through multispectral image fusion
Authors: Bautista Pinky Abarca , Yagi Yukako
Published: June 2021

Effective visualization of the different tissue structures in a stained tissue section is important in pathology diagnosis. Different staining protocols are utilized to impress contrasting colors to tissue structures of interest. Popular to routine staining is the H&E stain, which highlights the difference between the nuclei and cytoplasm areas. In this work, we present a multispectral image fusion method which improves the delineation between tissue structures that originally display similar H&E staining patterns. Multispectral images of H&E stained tissue sections were captured at a spectral resolution of 5nm from 400nm–720nm. Multispectral image fusion is implemented using two sets of principal component (PC) vectors: (i) PC vectors derived from the spectral transmittance samples of the various tissue components; and (ii) the PC vectors calculated from the spectral residual‐errors acquired by the tissue components in the spectral reconstruction. From the result of the multispectral image fusion we can readily observe the difference between tissue structures which originally share similar H&E staining patterns.
Ontogeny of sphenoidal, occipital, and temporal bone morphology in humans
Authors: Dalal Deepal , Smith Heather F.
Published: June 2021

Previous research suggests that the basicranium ossifies early in ontogeny and reflects phylogeny, but the process by which the various basicranial subregions develop is currently unknown. The 3D basicranial landmarks on the sphenoid, temporal, and occipital of 266 subadults and adults in seven populations were collected and compared. The seven populations were found to be significantly different in basicranial morphology in specimens over 5 years old, and those differences significantly reflected their published molecular distances. The differences in morphology significantly reflected genetic distances in childhood, but not adulthood. The occipital bone showed significant differences in morphology in children, and significantly reflected genetic distances; however, this pattern did not continue into adulthood. Temporal bone age categories reflected genetic distances if the Egyptian population was excluded. In sphenoid, the differences among groups were uncorrelated with genetic distances. These results indicate that the morphology of the entire basicranium reflects molecular distances and that this pattern becomes established early in ontogeny. However, the three basicranial regions develop along differing ontogenetic trajectories. Greater landmark density is preferable when using basicranial morphology to reconstruct population history.

Grant Funding Source: Midwestern University
Translating Carpal Biomechanics to Triangular Fibrocartilage Complex Injury: Significance of Ulnar Variance in Predicting TFCC Lesions
Authors: Baral Sumit , Adar Tony , Classie Justin Anthony , Marquez Samuel
Published: June 2021

Ulnar variance (UV) describes the cranio‐caudal position of the distal ulna in relation to the distal radius at the wrist. Neutral ulnar variance (ulna and radius articulating at the carpus at exactly the same level) transmits 20% of the compressive load across the wrist to the distal ulna and 80% to the distal radius. A +2.5mm positive UV has been shown to increase the ulnocarpal load to 40%. The triangular fibrocartilage complex (TFCC) acts as a cushion for the ulnar carpus and transmits axial loads from the wrist to the ulna in a longitudinal direction across the ulnar column of the wrist. Consequently, a positive ulnar variance implicates a greater risk of TFCC injury. This study sought to calculate the prevalence of traumatic TFCC injury in those with a positive UV versus those with a negative UV. 20 cadaveric wrists aged 50 – 90 were assessed for TFCC damage via gross examination and classified according to Palmer's schema. A plain film PA radiograph of each wrist was then taken to measure the degree of UV. Results showed that all wrists had degenerative Class II lesions of the TFCC, while traumatic Class I lesions of the TFCC were found in 69% of wrists with positive UV as compared to 0% of wrists with negative UV (p<0.05). Our investigation prompts further research exploring a “dose‐response” relationship between UV and TFCC damage as well as between the degree of TFCC damage and clinical symptoms or diminished quality of life.

Grant Funding Source: SUNY Downstate College of Medicine Alumni Association
Development of an effective in vitro epithelial cell infection model to study Edwardsiella tarda infections
Authors: Tenkink Byron Joel , Siame Bupe Anthony , Leung Ka Yin , Guttman Julian Andrew
Published: June 2021

Edwardsiella tarda is an enteric pathogen that infects both warm‐water fish and humans. While it is known that E. tarda utilizes syringe‐like secretion systems to deliver bacterially‐derived pathogenic proteins into macrophages during infection, little is known about E. tarda pathogenesis in epithelial cells. To enable the investigation of E. tarda infection mechanisms, we hypothesized that an effective and reliable E. tarda infection model could be developed by utilizing epithelial cells that originate from organs that are targeted during in vivo infections. To develop this in vitro model, isolates of E. tarda from various locations around the world were used to infect 4 different epithelial cell lines derived from fish, using various bacterial loads and durations of the infections. We found that E. tarda infected 10–15% of BF‐2 (Bluegill fry caudal trunk cells) when used at a multiplicity of infection of 10 for three hours at 30°C. The E. tarda infection rate was measured by counting infected cells per slide after fixation and visualization by fluorescence microscopy. Our results constitute the most effective E. tarda infection model ever developed and provides us the foundation to elucidate the strategies E. tarda utilize to cause disease.

Grant Funding Source: Natural Sciences and Engineering Research Council
Francisella tularensis does not alter ubiquitylated or SUMOylated proteins profiles during epithelial cell infections
Authors: Chua Michael , Guttman Julian A.
Published: June 2021

Francisella tularensis are intracellular bacteria that cause tularaemia, an infection that can lead to 60% mortality rates if left untreated. Although these microbes infect both macrophages and epithelial cells, the epithelial cell infections are known to be crucial for the disease process. A variety of bacteria alter the normal processes of post‐translational protein modifications that occur in healthy epithelial cells. These include the processes of ubiquitylation and SUMOylation. To begin to determine the host cell alterations that occur during Francisella‐based epithelial cell infections, we hypothesized that ubiquitylation and/or SUMOylation alterations to host proteins would be altered during the infections. To test this hypothesis, we analyzed protein immunoblots of two cell lines, HeLa and COS‐7, that we infected with Francisella for 24 hours. Comparison of uninfected and infected samples showed no significant differences in ubiquitin, SUMO1 or SUMO2/3 modifications to any host cell proteins. Through our study, we concluded that Francisella does not alter the host ubiquitin or SUMO post‐translational protein modifications as part of their disease mechanism.

This study was funded by CIHR.

Grant Funding Source: Canadian Institutes of Health Research
Environmental Estrogens Decrease Agrin‐Induced Acetylcholine Receptor Clustering
Authors: Renteria Sarah M , Grow Wade A
Published: June 2021

In addition to reproductive tissues, estrogen hormones target skeletal muscle. Studies have linked the decline of estrogen during menopause to sarcopenia, a loss in muscle mass and strength with aging. Steroid hormones including estrogens alter gene transcription and affect cell function. Human exposure to environmental estrogens that can mimic endogenous estrogens is increasing. Our lab uses the C2C12 cell culture model to investigate skeletal muscle development, specifically the events of myotube formation and acetylcholine receptor (AChR) clustering. C2C12 myoblasts proliferate and fuse into myotubes in cell culture. Clusters of AChRs form spontaneously on myotubes with motor neuron derived agrin increasing the frequency of AChR clustering. Our objective was to assess if environmental estrogens affect agrin‐induced AChR clustering. We tested four environmental estrogens that represent a range of distributions and toxicities: DDT, methoxychlor, estrone, and 17â‐estradiol. Our results demonstrate that individually and in combination environmental estrogens can decrease agrin‐induced AChR clustering. We conclude that environmental estrogens may interfere with skeletal muscle development.

Grant Funding Source: Sarah M. Renteria was supported by the Midwestern University Biomedical Sciences Program. Wade A. Grow was supported by Midwestern University intramural funds.
A Unique Branching Pattern of Celiac Trunk. Its Clinical and Embryological significance
Authors: Valentin Natalia , Veras Wilson R , Duarte Jhonathan , Vilaró José
Published: June 2021

The celiac trunk is the first ventral branch of the abdominal aorta and is the major blood supply to the supracolic abdominal compartment. Usually, three branches arise from the celiac trunk to supply this region, the splenic, the common hepatic and the left gastric arteries. During a routine dissection in the anatomy laboratory an extraordinary anatomical variation of the celiac trunk and its branches is found. The present case correspond of a celiac trunk, which gave off six branches. It gave off three classical branches, that is left gastric, splenic and common hepatic arteries. Beside these three branches, the right and left inferior phrenic arteries, and the superior posterior pancreaticoduodenal artery arose from the celiac axis. These anatomical variations were accompanied by a right gastric artery arising from the grastroduodenal artery instead of the common hepatic artery as describe in textbooks.

During normal development, both dorsal aortas give rise to many ventral segmental (omphalomesenteric) arteries. The ventral segmental arteries regress shortly after fusion of dorsal aortas. The dorsal aorta gives off segmental branches to the digestive tube (ventral splanchnic arteries), the mesonepheric ridge (lateral splanchnic arteries), and intersegmental branches to the body wall (somatic arteries). Anatomical variations of the coeliac trunk are due to developmental changes in the ventral segmental arteries. These ventral segmental arteries supply the yolk sac, allantois and chorion. Three ventral segmental arteries remain as coeliac trunk, superior mesenteric artery and inferior mesenteric artery. Anatomical variations in the branching pattern of the celiac trunk are of considerable importance in liver transplants, laparoscopic surgery, invasive radiological interventions and penetrating injuries to the abdomen. Knowledge of variation found in the present case is very useful to kept in mind to avoid serious vascular complications.
The myosin motor Myo1c is required for VEGF‐induced VEGFR2 delivery to the cell surface and angiogenic signaling
Authors: Choudhury Amit
Published: June 2021

Vascular endothelial growth factor receptor‐2 (VEGFR2) is a receptor tyrosine kinase expressed in endothelial cells and regulates angiogenic signal transduction processes during physiological and pathological conditions. Intracellular vesicular transport of VEGFR2 along endocytic and secretory transport pathways regulates turnover of this receptor at the plasma membrane (PM). Motor proteins of the myosin family regulate short range cargo trafficking along actin filaments. In the current study using primary human endothelial cells, we demonstrate that Myo1c, an unconventional class I myosin, regulates the availability of VEGFR2 at the PM. Myo1c co‐localizes with VEGFR2 and caveolin‐1 (Cav1) at the PM. VEGFR2 recruitment, and co‐localization with Myo1c and Cav1 at the PM occurs in response to VEGF‐A stimulation. Depletion of Myo1c reduces total and cell surface VEGFR2 that could be rescued by overexpression of wild type form of Myo1c. VEGF‐induced VEGFR2 delivery to the PM requires Myo1c, as depletion of Myo1c targets the receptor to the lysosomes for degradation. Depletion of Myo1c reduces VEGFR2 phosphorylation at Y1175 and also reduced activation of ERK1/2 and c‐Src that leads to reduced cell proliferation and cell migration, respectively, in response to VEGF. This study is the first report showing Myo1c as an important player for VEGF‐induced VEGFR2 delivery to the PM and angiogenic signaling.
Overexpression of MnSOD provides protection against NRTI‐induced endothelial dysfunction in a drug‐dependent manner
Authors: Glover Mitzi , Hebert Valeria , Xue Stephen , Thibeaux Taylor , Zavecz James H. , Dugas Tammy R
Published: June 2021

Nucleoside reverse transcriptase inhibitors (NRTI) currently used in HIV therapy have been linked to cardiovascular toxicity. Our prior studies have demonstrated alterations in mitochondrial function and upregulation of mitophagy in endothelial cells after acute treatment, with parameters returning to normal by 24 h. Treatment with coenzyme Q10 prevented this toxicity, suggesting a role for antioxidant supplementation. The current study utilized mice overexpressing MnSOD to determine the protective effects of this mitochondrial antioxidant enzyme. Measurements of plasma vasodilators, vasoconstrictors, and markers of oxidative stress, along with vessel reactivity experiments were used to assess protective effects. Results indicate that although wild‐type mice treated with either AZT or 3TC exhibited reduced vessel reactivity, overexpression of MnSOD only attenuated 3TC‐induced effects. Plasma markers of vascular function either correlated with this finding, or demonstrated MnSOD protection for both NRTI. These findings suggest differential mechanisms of toxicity of the 2 drugs. Supported by R01‐HL08242–0251.

Grant Funding Source: R01‐HL08242–0251
Age‐Dependent Myocardial Sheet Dysfunction from 3 to 16 months in Duchenne Muscular Dystrophy Mice (mdx) Defined by Diffusion Tensor Magnetic Resonance Imaging (DTI)
Authors: Cheng Ya‐Jian , Chen Junjie , Caruthers Shelton D. , Wickline Samuel A.
Published: June 2021

INTRO

Cardiomyocytes are organized in laminar structures (aka, sheets) that contribute to wall thickening/thinning. Diffusion tensor MRI (DTI) measures the sheet reorientation function in intact hearts. We have reported that hearts of 16‐mo old mdx mice exhibit abnormal Ca transients & Ca‐dependent sheet dysfunction at the base but not apex. Because Ca mishandling in isolated mdx cardiomyocytes is age dependent, we hypothesized that sheet dysfunction in intact mdx hearts may also be age dependent.

METHODS

Excised hearts from 3 & 16 mo old mdx and wildtype were prepared for Langendorff perfusion, and sequentially arrested in diastole then systole for DTI (Varian 11.7 T). The diastolic arrest was achieved with either normal [Ca] (1.2 mM, noted as NC) or low [Ca] (0.078 mM, noted as LC) cardioplegic solution to test the Ca dependency. The systolic arrest was achieved with barium.

RESULTS

Systolic |β| (defined below) was similar between WT and mdx, whereas diastolic |β| in mdx‐NC was lower for age matched WT‐NC at the base but not apex, indicating a regional failure to fully relax. Low [Ca] (mdx‐LC) perfusion restores the diastolic |β| to normal values at both ages. The Δ|β| (i.e., |β| of each group ‐ |β| of age matched WT‐NC) is 50% larger in 16 mo than for 3 mo old mice, indicating that Ca dependent sheet dysfunction is age dependent and regionally manifest despite the ubiquitous dystrophin deficiency.
Correlation between the expressions of the natriuretic peptide system and morphological changes in mouse heart during prenatal development
Authors: Li Terry Yantian , Tse M. Yat , Ventura Nicole M. , Croy B. Anne , Reifel Conrad W. , Pang Stephen C.
Published: June 2021

Natriuretic peptides, most noticeably ANP and BNP are synthesized by heart and play an important role in blood pressure regulation and fluid homeostasis. These hormones express during early fetal development and have a different expression pattern than that in adults. In addition, ANP is known to serve as a growth modulator in the adult heart. In this study we sought to determine the expression of ANP and BNP during fetal development in order to correlate them with morphological changes in the heart during this critical period of cardiac growth. The expression of ANP and BNP were studied using real‐time quantitative PCR (qPCR) at daily intervals from 12 to 19 gestational days (GD), as well as in neonatal and adult hearts. The expression of GATA4, a transcriptional factor of ANP and BNP, was also measured. In another series of mice, hearts were dissected for LM morphological analysis. Results showed that peak ANP expression was observed on 15GD; whereas expression levels of GATA4 and BNP were significantly increased after 12GD. Morphological analysis demonstrated that there was an increase in cardiomyocyte size between 14–15GD, which was coincided with the peak of ANP expression. We suggest that ANP may play a role in modulating cellular hypertrophy in the heart during prenatal development.

Grant Funding Source: Garfield Kelly Foundation at Queen's University, HSFC and CFI
SIRT2 Overexpression in Hepatocellular Carcinoma Mediates Epithelial to Mesenchymal Transition via Akt/GSK‐3β/β‐catenin Signaling.
Authors: Ko Ben Chi‐Bun , Chen Juan , Chen Anthony , Lai Paul B.S. , Huang Ailong
Published: June 2021

The yeast silent information regulator 2 (SIR2) is a histone deacetylase that utilizes NAD+ as a cofactor for its functions. In mammals, SIR2 is represented by seven homologues (SIRT1–7). We found that Sirtuin 2 (SIRT2) contributes to cell motility and invasiveness of HCC. SIRT2 is upregulated in HCC cell lines and in a subset of human HCC tissues (23/45). Up‐regulations of SIRT2 in primary HCC tumors were significantly correlated with the presence of microscopic vascular invasion (p=0.001), more advanced tumor stage (p=0.004) and shorter overall survival (p=0.0499). Functional studies by shRNA‐mediated suppression of SIRT2 expression in HCC cell lines revealed significant inhibition of motility and invasiveness. Depletion of SIRT2 also led to the regression of epithelial‐mesenchymal transition (EMT) phenotypes, whereas the ectopic expression of SIRT2 in the immortalized hepatocyte cell line L02 enhanced the expression of EMT markers, and promoted cell motility and invasiveness. Mechanistic studies revealed that SIRT2 regulates the deacetylation and activation of Akt, which subsequently impinges on GSK‐3β/β‐catenin signaling pathway to regulate EMT. Concordantly, we showed that there is a positive correlation between the level of SIRT2 and β‐catenin in human HCC. Our findings uncover a novel role of SIRT2 in HCC metastasis, and provide a rationale to explore the use of sirtuin inhibitors in HCC therapy.
Spontaneous salivary gland carcinomas in GON4l deficient mice
Authors: Gibson‐Corley Katherine Nicole , Simons Andrean L. , Lu Ping , Robinson Robert A. , Meyerholz David K. , Colgan John D.
Published: June 2021

We have described a Gon4l mutant mouse in which B cell development is impaired. When aged, these mice spontaneously develop salivary gland tumors at a rate of about 25%. These tumors were associated with the submandibular salivary gland and to a lesser extent with the sublingual and parotid glands. Histologically these masses had central cavitations containing necrotic cellular debris. Tumor cells varied from spindle to epithelial in shape with fewer cells exhibiting basaloid and clear cell differentiation. Tumor cells had a high mitotic rate, pleomorphism and invasion into adjacent salivary glands. These masses show striking similarity to human malignant salivary gland neoplasms which have myoepithelial and basal cell differentiation. Immunohistochemical staining revealed diffuse immunoreactivity for pancytokeratin (AE1/AE3), p63 and phosphorylated and total epidermal growth factor receptor (EGFR). CK5/6 immunostaining was noted in the majority of tumor cells, although was not evident in areas of marked pleomorphism. Alpha‐smooth muscle actin, S100, c‐kit and glial fibrillary acid protein were all immunonegative. Based on the morphological and immunohistochemical findings, we diagnosed these tumors as carcinomas with myoepithelial and basaloid cell differentiation. This novel murine model may be a valuable tool for studying human salivary gland neoplasia and EGFR regulation. This work was supported in part by NIH Grant R01 AI093737.
Radiosensitivity of Human Inducible Pluripotential Stem Cells (iPSCs)
Authors: Greenberger Joel S. , Houghton Frank D. , Zhang Xichen , Nimgaonkar Vishwajit L. , D'Aiuto Leonardo , Epperly Michael W.
Published: June 2021

Inducible human pluripotent stem cells, derived from fibroblasts, have been purposed for repair of tissue damage in various organs. iPSCs have displayed robust potential for providing reagent stem cells for tissue repair and regeneration. We evaluated the radiobiologic properties of a parent human fibroblast cell line and the iPSC derivative from plasmid transfection with OCT4, SOX2, NONOG and LIN28. We determined the radiosensitivity of iPSC cells compared to parental fibroblasts, and derivitive neural rosettes, prepared by growth of the iPSCs in 1% N2 supplement and bFGF. Cells of each cell type were irradiated to doses ranging from 0 to 2.5 Gy and the percent of apoptotic cells determined 24 or 48 hr after irradiation. The iPSC cells and rosettes were radiosensitive compared to fibroblasts: 43.6 ± 8.0 and 35.1 ± 3.9% apoptotic cells, respectively, compared to 1.9 ± 0.5% of fibroblasts cell at 24hrs after 2.5 Gy (p = 0.0261 and 0.0146, respectively). By 48 hr after irradiation, after significant cell loss, remaining iPSCs showed 18.0 ± 4.1 apoptosis compared to 7.1 ± 3.9% for fibroblasts and 30.9 ± 1.7% for rosettes. Five days after 2.5 Gy irradiation iPSCs showed a 98.5% decrease in cell number compared to 78% for fibroblasts (p = 0.0174). Elucidation of the mechanism of radiosensitivity of iPSCs may reveal a specific role for expression of one or more of the 4 transgene products in the parental fibroblast cell line.

Supported by NIAID /NIH grant U191A168021–01 from NIAID
Comparison of 3D Reconstructive Technologies used for Morphometric Research and the Translation of Knowledge using a Decision Matrix
Authors: Martin Charys M , Roach Victoria A , Nguyen Ngan , Rice Charles L , Wilson Timothy D
Published: June 2021

The use of three‐dimensional (3D) models for education and measurement has become more prevalent, which has resulted in an increase in 3D reconstructive software programs. These programs differ in reconstruction concepts, operating system requirements, user features, cost, and no one program has emerged as the standard. The purpose of the current study was to conduct a systematic comparison of three widely available 3D reconstructive software programs, Amira, OsiriX, and Mimics, with respect to the software's ability to be used in morphometric research and education to translate morphological knowledge. Cost, system requirements, and inherent features of each program are compared. A novel concept selection tool, a decision matrix, was employed to objectify comparisons of usability of the interface, quality of the output, and efficiency of the tools. Findings indicate that Mimics was the best‐suited program for construction of 3D anatomical models and morphometric analysis, but for creating a learning tool the results were less clear. OsiriX was very user‐friendly; however, it had limited capabilities. Conversely, although Amira had endless potential and could create complex dynamic videos, it had a challenging interface. These results provide a resource for morphometric researchers and educators to assist the selection of appropriate reconstruction programs when starting a new 3D modelling project.

Grant Funding Source: Departmental
Anatomical validation of diffusion tensor imaging (DTI)
Authors: Liu Robert Ming , Hageman Nathan , Yang Gloria , Cheng Nathan , Chan Kelly , Liu Erica , Ortiz Jake , Honarpisheh Helen , Wong Anita , Stark M. Elena , Dong Hongwei , Vinters Harry , Toga Arthur , Wisco Jonathan
Published: June 2021

The Magnetic Resonance Image (MRI) based Diffusion Tensor Imaging (DTI) technique reveals brain white matter architecture in vivo by measuring the direction of the diffusion of water molecules along the anisotropic fiber bundles. Validation of DTI at present is limited to qualitative comparisons of previously known connections in the common normal human brain. Anatomical validation of DTI through histological analysis ensures that each image is representative of the inherent white matter architecture, and improves algorithms that create a tractographic image from the tensor signals in each voxel of the image. Human brain samples were imaged at 3 Tesla (T) (2.5 mm voxels) followed by sectioning and histologic processing of the tissue. Results comparing the histologic data to the imaging show good correspondence in white matter areas between both fiber integrity and axonal orientation. Vector coordinates drawn from the tissue image match the principal fiber direction derived from the probability density function of the corresponding image voxel. We anticipate that this will provide a valuable resource for the validation of DTI methods.

Grant Funding Source: NIH/NIA 1 R21 AG037843‐02, NIH/NCRR P41 RR013642‐12S1, Translational Research Fund (UCLA), and the McGinty Family Foundation
Effects of heroin dependence on the expression of DBH and CCK in the VTA and NAc regions of rat brain
Authors: Li Yi‐xin , Liang Wen‐mei
Published: June 2021

To investigate the effect of the heroin addicts on rat's DβH‐, CCK‐immunoreactive cells in the ventral tegmental area(VTA) and nucleus accumbens(NAc) regions, and exploring the effect of heroin dependence on regulation of dopamine release and self‐protective mechanism in brain. Adulthood 55 male SD rats were randomly divided into normal control group(NCG), saline control group(SCG) and the heroin‐dependent group(HDG). Heroin‐dependent rat models were set up by the way of subcutaneous injection of heroin and brain tissues are excised on the 10th, 17th, 24th, 31th, and 38th days after models are set up. Immunohistochemical staining was performed to test the expression of DβH and CCK in the VTA and NAc regions. The expression of DβH and CCK was observed as brown yellow particle in the cytoplasm of neurons in the VTA and NAc regions. The expression of DβH‐ immunoreactive cells is down‐regulated, while the expression of CCK‐immunoreactive cells is up‐regulated in the VTA and NAc regions from HDG rats than that from SCG and NCG rats at different time points. The results suggest that the secretion of DβH is inhibited during heroin dependence, and it might be one reason causing dopamine increase in VTA and NAc. The increased secretion of CCK in VTA and NAc might be self‐protective mechanism in brain.

Grant sponsor(s): NSFC(Natural Science Foundation of China)

Grant number(s):81060110

Grant Funding Source: Natural Science Foundation of China
Histological validation of iron as an imaging biomarker for amyloid beta and tau depositions in Alzheimer's Disease
Authors: Salin Ashley , Hansen Megan Salin , Barzee Brigham , Stone Megan , Bridgewater James , Kavafyan Talar , Steed Kevin , Stark Elena , Dong Hongwei , Toga Arthur , Vinters Harry , Wisco Jonathan
Published: June 2021

At present, no one can diagnose a patient with Alzheimer's disease (AD) until postmortem examination. Analyzing spatial overlap of non‐heme iron, tau protein, and amyloid beta (Aβ) depositions in consecutive tissue slices of the hippocampus of human subjects would ultimately facilitate characterization and detection of diseases, such as AD, through MRI. Iron is readily visible in T2* MRI, and has been shown to be spatially correlated with Aβ plaques. Neurofibrillary tangles of tau protein are also associated with AD. In this study, we verified iron deposition to pathology of the hippocampus. We used Olympus OlyVIA software to capture virtual microscopic images, and Adobe Photoshop to register serial stained sections of iron, tau, and Aβ. Our initial results indicate spatial similarities between all three components. We plan to perform this entire procedure for the hippocampus in hopes of providing useful diagnostic research relative to AD. Pathological validation through these images will clarify the configuration of these probable biomarkers and their specificity to AD and other neurodegenerative diseases.

Grant Funding Source: NIH/NIA 1 R21 AG037843‐02, NIH/NCRR P41 RR013642‐12S1, Translational Research Fund (UCLA), and the McGinty Family Foundation
Neurotensinergic periaqueductal gray‐rostral ventromedial medulla‐spinal dorsal horn pathway
Authors: Li Yunqing
Published: June 2021

Aim

To investigate the neurotensin (NT)‐containing periaqueductal gray (PAG)‐rostral ventromedial medulla (RVM)‐spinal dorsal horn (SDH) descending pathway, which has been shown functionally antinociceptive in nature.

Methods

Fluoro‐gold (FG) or horseradish peroxidase (HRP) retrograde and biotinylated dextran amine (BDA) anterograde tracing methods and immunohistochemistry combined with tracing methods were used in the rat.

Results

After FG and BDA was injected into SDH or PAG, respectively, and combined with immunohistochemistry for NT and NT receptor 1 (NTR1), we found that NTR1‐immunoreactive (IR) neurons in the RVM received NT‐IR projections from PAG and also sent projections to the SDH. Then under electron microscope, in the RVM, we further found: (a) NT‐IR terminals made synapses on HRP‐labeled dendrites showing NTR1‐immunoreactivities; (b) BDA‐labeled terminals made synapses on HRP‐labeled dendrites exhibiting NTR1‐immunoreactivities; (c) BDA‐labeled terminals containing NT made synapses on HRP‐labeled dendrites.

Conclusions

PAG projecting axon terminals contain NT and form asymmetric synapses on NTR1‐IR neurons in the RVM which sent descending fibers to the SDH. They might be involved in the NT‐induced analgesia.

This project was supported by grants from the National Natural Science Foundation of China (Nos. 30771133, 30971123, 31010103909).
Ameliorative role of Alpha lipoic acid (ALA) in sodium arsenite (NaAsO2) induced neurotoxicity in developing rat cerebellum
Authors: Dhar Pushpa , Kaushal Parul , Mehra Raj D
Published: June 2021

The present study was designed to evaluate the effects of ALA on arsenic (As) induced alterations in developing rat cerebellum. Pregnant Wistar rats, obtained from Experimental Animal Facility (EAF) after ethical clearance from Institutional Ethical Committee of AIIMS were maintained on a 12 hr light‐dark cycle with ad libitum access to food and water. The day of birth of pups was designated as postnatal day (PND) 0. Mother reared pups were divided into control (normal and sham) and experimental groups receiving NaAsO2 alone or along with ALA from PND 1 to 21. Behavioral parameters (tail hung, negative geotaxis and water maze) were carried out on PND 21. Animals were sacrificed on PND 22 and the fresh cerebella were processed for estimation of antioxidant glutathione (GSH) and Lipid peroxidation product malondialdehyde (MDA) levels. Cerebella from perfusion fixed animals were processed for paraffin embedding and Cresyl Violet stained sections were observed for morphological features. Immunohistochemical expression of synaptophysin (Syp) and Myelin basic protein (MBP) was studied in cryocut sections. The observations of present study revealed alteration in cerebellar morphology and behavioral dysfunction in rat pups exposed to NaAsO2.Co‐administration of ALA resulted in partial reversal of As induced decline in the behavioral parameters, decreased GSH levels, elevated MDA levels, disruption of cerebellar cytoarchitecture and decrease in Syp and MBP expression suggesting thereby protective role of ALA in NaAsO2 induced neurotoxicity.

DST INSPIRE Fellowship, Govt. of India (110053) to Parul Kaushal is acknowledged.
Taurine Protection against Endoplasmic Reticulum Stress in an Animal Stroke Model of Cerebral Artery Occlusion and Stroke Related conditions in Primary Neuronal Cell Culture
Authors: MohammadGharibani Payam , Modi Jigar , Menzi Janet , Prentice Howard , Wu Jang Yen
Published: June 2021

Taurine has been shown to provide protection against neurological diseases, such as Huntington's and stroke. We have shown that taurine can provide effective protection against Endoplasmic Reticulum (ER) stress induced by excitotoxicity or oxidative stress in PC12 cell line or primary neuronal cell cultures. In this study, we employed hypoxia/reoxygenation conditions for primary cortical neuronal cell cultures as well as the model of rat focal Middle Cerebral Artery Occlusion (MCAO). Using taurine greatly increased cell viability in primary neuronal cell culture and decreased the infarct area of sections at 6mm from 47.42±9.86% to 26.76 ± 6.91% in the MCAO model. Furthermore levels of the ER stress protein markers GRP78, Caspase12, CHOP and p‐IRE‐1 which were markedly increased in both the in vitro and in vivo models, significantly declined after taurine administration, suggesting that taurine may exert neuroprotection functions in both models. Moreover, taurine could downregulate the ratio of cleaved ATF6 and full length ATF6 in both models. In the animal model of stroke, taurine induced an upregulation of the Bcl‐2/Bax ratio and downregulation of Caspase‐3 protein activity indicating that it attenuates apoptosis in the core of the ischemic infarct. Our results show not only taurine elicits neuroprotection through the activation of the ATF6 and the IRE1 pathways, but also it can reduce apoptosis in these models

Grant Funding Source: 09KW‐11, Department of Health, State of Florida