bluuebunny/crossref_metadata_2025 huggingface dataset:
The role of sphingolipid metabolism in lifespan and healthspan extension in C. elegans
Authors: Radeny Joycelyn , Chan Jason
Published: June 2021

Multiple advancements in research have led to a better understanding of the aging process. However, many metabolic pathways that promote healthy aging are still unknown. Here, we investigate the bioactive sphingolipid metabolism pathway and its roles in regulating lifespan and healthspan, which is the average period of one's life in good health. Sphingolipids are involved in cellular signaling and known to promote cell proliferation, secretion, stress response and survival. The sphingolipid pathway is composed of sphingomyelin, ceramide, sphingosine and sphingosine‐1‐phosphate (S1P). Ceramide synthases (CER synthases) mediate the conversion of ceramide to sphingosine while sphingosine kinase (SPHK) converts sphingosine to S1P. Using C. elegans, we previously found that mutations in sphk‐1/SPHK and hyl‐2/CER synthase result in decreased lifespan, suggesting that sphingolipids play a role in lifespan extension. However, it is unknown whether the sphingolipid genes act in known cellular pathways that mediate aging. Therefore, we tested the genetic interactions between sphingolipid enzymes and known models of aging, including the insulin like signaling (ILS) pathway that has been linked to lifespan extension. For this, lifespan analyses were performed on daf‐16/FOXO, which has a shortened lifespan phenotype, coupled with knockdown of sphingolipid metabolism genes. DAF‐16 is a downstream transcription factor target of DAF‐2/insulin‐like growth factor receptor and has important roles in mediating the gene expression of stress response. We observed that daf‐16 is epistatic to sphingolipid mutants for lifespan phenotypes, as indicated by similar short lifespan phenotypes of double knockout of daf‐16;hyl‐2 and daf‐16;sphk‐1compared to mutants with a single knockout of daf‐16. Thus, sphingolipids may act upstream of DAF‐16 to mediate stress response genes and lifespan. Future experiments will analyze transcriptional reporters for DAF‐16, and known stress response genes SOD‐3/superoxide dismutase, to determine whether cellular stress response mechanisms occurs normally in hyl‐2 and sphk‐1mutants under induced oxidative stress. An analysis of stress response will further enhance our understanding of the interaction between sphingolipids and known pathways that promote lifespan and healthspan.

Support or Funding Information

Research reported in this publication was supported by the National Institute On Aging of the National Institutes of Health under Award Number R15AG052933.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anthropometry, Energy Metabolism and Nutritional Intake of Girls with Adolescent Idiopathic Scoliosis
Authors: Normand Emilie , Franco Anita , Parent Stefan , Moreau Alain , Marcil Valerie
Published: June 2021

Purpose of the study

Adolescent idiopathic scoliosis (AIS) is a tridimensional spine deformation of unknown cause that affects 2–3% of children and mostly girls. Studies have suggested that girls with AIS are taller and leaner than controls, but more studies are needed to confirm these results. While differences in energy metabolism and nutritional intake could explain these discrepancies, they have been poorly studied in AIS. This study aims to thoroughly characterize the anthropometric, metabolic and nutritional profiles of girls affected by AIS and to compare them to age‐matched healthy controls.

Methods

20 girls with AIS and 20 healthy controls are being enrolled. Anthropometric parameters (weight, standing and sitting height, body mass index (BMI), waist circumference and arm spam) are measured. Fasting glucose, insulin and lipid profile (total cholesterol, LDL‐cholesterol, HDL‐cholesterol, triglycerides and apolipoprotein B) are assessed. Bone mineral density and body composition are determined by osteodensitometry scan and questionnaires are administered to assess food intake and physical activity level.

Results

To this day, 17 AIS (14.9 ± 1.9 yrs, mean Cobb angle of 27.2 ± 10.0°) and 12 healthy girls (15.2 ± 1.6 yrs) have completed the study. Compared to controls, we observed in AIS girls tendencies for lower BMI (BMI‐for‐age z‐score: −0.50 ± 0.90 vs. 0.09 ± 1.20, P=0.3669), fat percentage (−15%, P=0.1213), waist circumference (−7%, P=0.0706) and bone mineral density (−8%, P=0.0217). There was no difference in fasting glucose (4.98 ± 0.27 vs. 4.84 ± 0.27 mmol/L, P=0.1852) and insulin (52.46 ± 19.87 vs. 54.12 ± 19.41 ρmol/L, P=0.8365) between groups. A tendency for a perturbed lipid profile was also found in AIS girls with higher triglycerides (18%, P=0.2866), LDL‐cholesterol (16.5 %, P=0.0306), apolipoprotein B (19%, P=0.0071) and total cholesterol (12%, P=0.0374). 47% of AIS girls had at least one abnormal lipid value. Calorie intake and physical activity levels were similar between groups. However, compared to controls, there was a tendency for higher caloric balance (consumption vs. needs) in AIS girls (+60.5 ± 417.7 vs. −223.4 ± 625.7 kcal, P=0.1706). AIS girls and controls had similar vitamin D (4.60 ± 2.30 vs. 5.40 ± 3.01 μg, P=0.4469) and calcium intake (954.78 ± 326.20 vs. 928.78 ± 385.12 mg, P=0.8509).

Conclusion

: Our pilot study revealed tendencies for different anthropometrical features, lipid profile and energy balance in AIS. Increasing the number of participants might confirm these results.

Support or Funding Information

Fondation Yves Cotrel‐Institut de France

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ishige okamurae Extract Reduces Obesity in High Fat Diet‐Induced Obese Mice
Authors: Seo Young‐Jin , Song Ji‐Hyeon , Chei Sungwoo , Lee Kippeum , Jin Heegu , Oh Jae‐Hoon , Oh Hyun‐Ji , Lee Boo‐Yong
Published: June 2021

Obesity is caused by an imbalance of energy intake and physical activity. A number of pharmaceutical agents exist to reduce body weight‐related symptoms of obese patients. Ishige okamurae is brown algae and has anti‐oxidant and anti‐inflammatory activities. However, it has not been reported that Ishige okamurae has anti‐obesity property. In our study, we identified that Ishige okamurae extract (IOE) suppresses high fat diet (HFD)‐induced obesity through modulation of adipogenesis and lipolysis. IOE fed mice decreased body and WAT weight. The expression of adipogenic factors such as CCAAT/enhancer binding protein alpha (C/EBPα) and peroxisome proliferator‐activated receptor gamma (PPARγ) were decreased by IOE. Additionally, IOE up‐regulated expression of lipolysis related enzymes such as adipose triglyceride lipase (ATGL) and hormone‐sensitive lipase (HSL). Therefore, our results suggested IOE suppresses adiposity through reduction of adipogenic and enhance of lipolytic‐related factors.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A mechanism for anesthetics through membrane lipids
Authors: Pavel Mahmud Arif , Petersen E. Nicholas Arif , Lerner Richard A. , Hansen Scott
Published: June 2021

Since their discovery more than 150 years ago, anesthetics were speculated to target the cellular membranes. Hitherto, there is no plausible mechanism that explains how anesthetics perturbation on membrane could to lead to an effect. Here we show that the volatile anesthetics chloroform and isoflurane activate TREK‐1 channels through disruption of ordered lipid domains (rafts). Employing super resolution microscopy, we found that raft disruption by anesthetics cause the raft‐localized enzyme phospholipase D2 (PLD2) to translocate and activate TREK‐1. A catalytically dead PLD2 significantly decrease the anesthetic‐specific TREK‐1 currents. Additionally, transfer of the PLD2 binding‐site to an anesthetic‐insensitive channel, TRAAK, results in anesthetic‐sensitive currents. The general anesthetics chloroform, isoflurane, diethyl ether, xenon, and propofol were all found to activate PLD2 in cellular membranes indicating a broadly applicable mechanism. We propose a two‐step model of anesthetic TREK‐1 activation. First, inhaled anesthetics disrupt lipid rafts. Second, translocation and PLD2‐dependent production of anionic lipid activate associated channels. This model suggests anesthetics could indirectly activate ion channel in contrast to direct binding models.

Support or Funding Information

This work was supported by a Director's New Innovator Award (1DP2NS087943‐01 to S.B.H.) from the NIH, a graduate fellowship from the Joseph B. Scheller & Rita P. Scheller Charitable Foundation to E.N.P. We are grateful to the Iris and Junming Le Foundation for funds to purchasea super‐resolution microscope, making this study possible.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The biological function of PIP2 clustering in enzyme activation and ion channel regulation
Authors: Hansen Scott B. , Pavel Mahmud Arif , Petersen E. Nicholas
Published: June 2021

The signaling lipid phosphatidylcholine 4,5 bisphosphate (PIP2) clusters with proteins in the plasma membrane away from saturated domains formed by GM1 lipids and cholesterol (GM1 domains or ‘lipid rafts’). The lipid metabolizing enzyme phospholipase D2 (PLD2) localizes to both lipid raft and PIP2 clusters creating a competition for localization. We show that disruption of GM1 domains by mechanical force or inhaled anesthetics causes PLD2 to translocate to PIP2 clusters where it activates twik related potassium (TREK‐1) channels giving rise to the channel's mechano and anesthetic sensitivity. We conclude cells have evolved to (i.) sense and respond to disruption of lipid order and (ii.) utilize lipid order to dynamically regulate enzymes and ion channels through substrate presentation.

Support or Funding Information

This work was supported by a Director's New Innovator Award (1DP2NS087943‐01 to S.B.H.) from the NIH, a graduate fellowship from the Joseph B. Scheller & Rita P. Scheller Charitable Foundation to E.N.P. We are grateful to the Iris and Junming Le Foundation for funds to purchase a super‐resolution microscope, making this study possible.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Glycosaminoglycan Recognition of Neutrophil‐Activating Chemokines
Authors: Nagarajan Balaji , Sepuru Krishna Mohan , Rajarathnam Krishna , Desai Umesh R
Published: June 2021

Neutrophil‐activating chemokines are known for their fundamental and diverse role in recruiting immune cells to the site of inflammation/infection. Extracellular glycosaminoglycans (GAGs), especially heparan sulfate, sequester chemokines and engineer a concentration gradient to regulate the immune cell trafficking. Not much is known about the structural and dynamical aspects of chemokine – GAG interactions. We have studied the interaction of several human and mouse chemokines, including CXCL1, CXCL2, and CXCL5 with heparin/heparan sulfate oligosaccharides using computational (modeling and dynamics) and biophysical (NMR & ITC) techniques. These studies have led to fine atomistic level understanding of GAG recognition of chemokines. First, our studies show that even though these chemokines are structurally homologous, they exhibit striking differences in their recognition of GAGs. Diverse binding geometries have been observed in which a GAG chain may bind each monomer present in a chemokine dimer or a GAG chain may bind two non‐overlapping sites by spanning across the dimer interface. Second, molecular dynamics simulations show large concerted motions of the Lys/Arg side chains bound to sulfate/carboxylates of GAGs, which imply translational motion of a GAG chain over the binding interface leading to considerable plasticity in binding. At the same time, a GAG chain may have restricted motion arising from specific interactions with a group of core amino acid residues. Third, a large number of hydrogen bond contributing residues occur in chemokines, e.g., in N‐loop, 40s turn, β3 strand and C‐helix, which provide strong/weak interactions to stabilize the complex. Interestingly, GAG recognition of chemokines is also brought about by water‐mediated interactions between amino acid side chains and sulfates/carboxylates, which introduce additional stability to GAG's dynamical motion. Fourth, relative importance of individual amino acid residues correlate well with experimental observations, thus implying that computational trajectories can be reliably used to understand GAG–chemokine systems. In addition, the computational trajectories can lead to identification of preferred chain length, mode of binding, and dynamical motions of GAGs in recognition of neutrophil‐activating chemokines. Our work contributes fundamental understanding on the structural biology of GAG–chemokine interactions.

Support or Funding Information

This work was supported in part by grants from the NIH including HL107152, HL090586 and HL128639

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Furin Inhibition as a Mechanism to Reduce Aberrant ENaC‐Mediated Sodium Transport and Rehydrate the Airways in Cystic Fibrosis Lung Disease
Authors: Douglas Lisa E.J. , Reihill James A. , Ho Melisa W.‐Y. , Axten Jeffrey M. , Martin S. Lorraine
Published: June 2021

The epithelial sodium channel (ENaC) is found in a number of tissues including lung epithelia and is activated through proteolytic cleavage of its α and γ subunits. Dysregulation of ENaC, secondary to the loss of functional CFTR in cystic fibrosis (CF), leads to increased Na+ absorption which contributes to airways dehydration, thickened mucus and impaired mucociliary clearance (MCC). Pro‐protein convertases such as furin are implicated in the activation of a distinct pool of ENaC as it passes through the biosynthetic pathway (1). Furin cleaves α‐ENaC at two sites releasing a small inhibitory peptide from the extracellular loop, partially activating the channel, and subsequently cleaves γ‐ENaC. These actions ‘prime’ the channel for cleavage at a secondary site on the γ subunit by cell‐surface channel activating proteases such as prostasin, trypsin and neutrophil elastase (2–4). Furin, therefore plays a key role in the proteolytic regulation of ENaC‐mediated sodium absorption and presents a promising therapeutic target for CF lung disease.

The aim of this first‐in‐kind study was to determine the effect of two chemically different, highly selective furin inhibitors (BOS‐981 and BOS‐318) on ENaC signaling and airway surface liquid (ASL) height, using primary human CF bronchial epithelial cells (HBEs).

Fully differentiated HBEs homozygous for F508‐del, grown at air‐liquid interface, were used to determine ENaC activity via equivalent current (Ieq) readings obtained using a 24‐well Transepithelial Current Clamp (TECC‐24) system. ASL height was measured using confocal microscopy.

We report BOS‐981 and BOS‐318 to be potent inhibitors of furin. Both compounds, applied basolaterally, significantly reduced ENaC‐mediated Na+ absorption as measured by an acute reduction in amiloride‐sensitive Ieq after immediate treatment, which was sustained after longer (48 hr) exposure. The compounds also protected ENaC against cell surface activation by neutrophil elastase and CF sputum supernatant. Furthermore, the reduction in ENaC activity after a 48 hr treatment correlated with an increase in ASL height.

These ground‐breaking results demonstrate that highly selective inhibition of furin by the novel inhibitors BOS‐981 and BOS‐318 presents a promising mechanism for the reduction of aberrant ENaC‐mediated Na+ absorption. This strategy has the potential to rehydrate the airways, restore effective MCC and reduce the devastating pulmonary decline observed in CF.

Support or Funding Information

This work was supported by funding from the MRC Proximity to Discovery: Industry Engagement Fund

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The selective BCL‐2 inhibitor ABT‐199 induces cell death via the intrinsic apoptotic pathway in canine B‐cell and T‐cell lymphoma cell lines
Authors: Meichner Kristina , Gogal Robert M.
Published: June 2021

The B cell lymphoma 2 (BCL‐2) family of proteins are critical regulators of apoptosis, acting as either activators or inhibitors of the intrinsic apoptotic pathway. Evasion of apoptosis by shifting the balance toward anti‐apoptotic proteins including BCL‐2 is an established mechanism of cancer cells. Lymphoma is a leading cause of cancer‐related deaths in dogs and there is evidence that BCL2 is overexpressed and can contribute to a therapy‐resistant phenotype. A selective BCL2 inhibitor, ABT‐199, shows clinical activity in human lymphoid malignancies but the effect in canine lymphoma is unknown. This study evaluated the effects of ABT‐199 on cell growth and different stages of apoptosis in two canine B‐cell lymphoma (CLBL‐1, 17–71) and three canine T‐cell lymphoma (OSW, Ema, Nody) cell lines. Cells were cultured for 72h in the presence of ABT‐199 at increasing concentrations to establish half maximal inhibitory concentrations (IC50). Nody (IC50: 0.49 μM), CLBL‐1 (2.72 μM), Ema (3.53 μM), and OSW (3.6 μM) were more sensitive to anti‐proliferative effects of ABT‐199 than 17–71 (12.93 μM). ABT‐199 rapidly induced key features of apoptosis including loss of mitochondrial membrane potential, increased caspase−3/−7 activity, and externalization of phosphatidylser‐ine. This effect was abrogated in cells co‐cultured with the pan‐caspase inhibitor Z‐VAD(OMe)‐FMK. The responses to ABT‐199 treatment are currently correlated with expression levels of BCL‐2 family members. These findings support that ABT‐199 induces cytotoxicity via induction of the intrinsic apoptotic pathway in canine lymphoma cell lines and that ABT‐199 shows promise to overcome drug resistance and to improve outcome in dogs with lymphoma.

Support or Funding Information

This study was supported by the University of Georgia VMES Research Grant 2926GR207002.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effects of Gossypol on Apoptosis‐Related Gene Expression in Racially Different Triple‐Negative Breast Cancer Cells
Authors: Messeha Samia , Mendonca Patricia , Soliman Karam F.A.
Published: June 2021

Apoptosis is a gene‐directed mechanism that regulates cell proliferation and maintains homeostasis. However, the uncontrolled apoptotic process can lead to several pathological conditions such as tumorigenesis, and cancer metastasis. In the current study, the apoptotic effect of the natural polyphenol compound gossypol (GOSS) was investigated in the triple negative breast cancer (TNBC) cells. The effect of GOSS was evaluated in two cell lines representing Caucasian Americans and African Americans: MDA‐MB‐231 (MM‐231) and MDA‐MB‐468 (MM‐468), respectively. In both cell lines, a similar response was noticed in both cytotoxicity and proliferation studies. However, MM‐468 cells were 3‐fold more sensitive to the apoptotic effect of the compound that was accompanied by a longer delay in the colony formation. Furthermore, GOSS was found to alter the expression of many apoptosis‐related genes in our mRNA expression investigation. The compound significantly upregulated GADD45A, TNFRSF9, and BNIP3 in MM‐231 cells. Similarly, GADD45A and BNIP3 were upregulated in MM‐468. A significant finding observed in this study is the profound 159‐fold increase in the TNF gene expression that was found in MM‐468 cells. Moreover, the apoptosis‐suppressor gene, BIRC5 was significantly repressed by more than 90% in both cell lines as well as DAPK1 in MM‐231 and TP73 in MM‐468 cells. In conclusion, the data obtained in this study can provide a more understanding of the GOSS‐induced apoptosis effect and enhance the importance of this polyphenol compound as targeted towards BC treatment, particularly in African American women.

Support or Funding Information

This research was supported by NIH NIMHD Grants G12 MD007582 and P20 MD 006738

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
"Incidence of Intracranial Pathologies in Acute Ocular Motor Mononeuropathies Suggests Preferred Practice Pattern"
Authors: DelGiodice Michael John
Published: June 2021

Imaging of the brain by magnetic resonance enables visualization of intracranial structures involved in the pathogenesis of acute, isolated ocular motor nerve palsies. However, a preferred practice pattern has yet to be determined. Meta‐analysis of prospective data, involving acute ocular motor mononeuropathy, identifies brain pathology with an incidence of 8.8%. Neuroimaging studies were associated with neoplasm, infarction, hemorrhage, aneurysm and demyelination. The recommendation is to obtain magnetic resonance imaging for acute ocular motor mononeuropathies, irrespective of neurologic isolation, diabetes and cardiovascular disease. These findings will be of value for clinicians seeking to adopt a preferred practice pattern.

Support or Funding Information

USAT College of Medicine

Associated Eye Physicians

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evaluation of the oncolytic ability of Theiler's murine encephalomyelitis virus in glioblastoma cell lines GL26 and GL261
Authors: Grey Devon , Freundt Eric C.
Published: June 2021

Glioblastoma multiforme (GBM) is type of brain cancer for which there are no effective treatments. Oncolytic viruses, such as poliovirus, are currently being tested as potential therapeutics in GBM. We recently found that Theiler's murine encephalomyelitis virus (TMEV), a relative of poliovirus, specifically downregulates expression of four transcription factors, known as OLIG2, POU3F3, SOX2, and SALL2 that are required for proliferation of GBM cells. However, these studies were done in stem‐cell derived oligodendrocyte progenitor cells and it is not known if TMEV can disrupt expression of these genes in GBM cells. In this study, we have infected the glioma cell lines GL26 and GL261 with two strains of TMEV, known as DA and GDVII and evaluated infection efficiency by immunofluorescence microscopy. While glioblastoma cells were susceptible to both strains, DA infected with greater efficiency than the GDVII strain. These differences were not observed in a fibroblast cell line, BHK 21 cells, which were equivalently infected by both strains. Chimeric viruses were used to determine whether strain‐specific differences in infectivity were due to capsid proteins of viral nonstructural proteins. Additionally, we assessed the oncolytic capacity and the ability of TMEV to downregulate OLIG2 in GBM cells. These results suggest that TMEV serves as a model to understand how picornaviruses can limit GBM proliferation.

Support or Funding Information

DG was supported by the Office of Undergraduate Research and Inquiry Student Research Grant

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Physical Therapy and Regenerative Medicine: Current Knowledge and Future Prospects
Authors: Pounders Vance , McArthur Libby , Rinehart Julian , McQueen Ryan , Raynes Edilberto , Roca‐Menchavez Melinda
Published: June 2021

Disorders affecting the central nervous system (CNS) leave patients with life‐altering disabilities. Injuries that occur in the brain and spinal cord are difficult to treat due to the lack of neuroregeneration in the CNS. Research demonstrates that regenerative medicine and physical therapy techniques can provide structural and functional improvements; however, there is limited research evaluating the combined approach. Regenerative medicine can mitigate the anatomical and physiological destruction caused by a CNS injury; while, physical therapy techniques can help the body restore normal function. The purpose of the study was to evaluate the efficacy of combination therapy using regenerative medicine and physical therapy and its impact on therapeutic outcomes for patients with CNS disorders. Using PUBMed, Google Scholar, and other databases, a systematic review was conducted and ranked using the Oxford Levels of Evidence. Based on the gathered evidence, we surmised that combination of regenerative medicine and physical therapy approaches could facilitate treatment of CNS injuries and lesions.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Telling the Truth About Caner is a Challenge
Authors: Markovic Olivera , Markovic Nenad
Published: June 2021

Cancer is our neighbor and is a part of our society. It has been here from the beginning of history and is still present hiding the truth about itself and being untouchable in its malevolent version: the unstoppable growth of abnormal cells attacking the body until it succumbs to the aggression.

Removing all malignant cells from the body is a challenge because it is the only real cure of the affected person. But, it can be done only if the disease is detected in precancerous phase, cancer in situ, and invasive cancer in clinical stage 1A. All more advanced cancers, notwithstanding the therapy employed, have a much worse prognosis. Medicine is there only to postpone the ultimate fatal outcome.

But, this is the same with life itself. It is the blessing to have it, but the challenge to live it. Cancer is the curse of our life. Therefore we have to fight against any carcinogenic and cancer promoter factor in our environments, to search for cancer indicators via surveys among healthy peoples, to put under control the groups at risk, and to conduct regular screening for detection of early signs of cancer as diligently as it has been done in the case of cervical cancer.

In this fight, medicine has made many advantages but the cancer is still present. Only a small fraction, the cervical cancer natural history has been altered due to detection of an early suspect for cancer lesions and their removal. The success was achieved in the US where Pap test has helped American health care providers to reduce its mortality rate for 80% and to reverse the curves of cancer prevalence and mortality from increasing pattern to decreasing.

This success was achieved in a group of 50 million women at risk, where 10% were found as suspect, subject to further diagnosis, only 1 million was confirmed as carrying on cancer indicators, and 40,000 lesions removed. Still, due to errors in sampling specimens, diagnostic errors or ineffective therapies, 12,000 joined the prevalence and 4,000 died per annum.

Worldwide, this concept failed. The global outreach is below 20% of women at risk, and curves of cervical cancer prevalence and mortality are still on rise. What can be done? It was our humble challenge for more than 30 years of research. Finally, a light started to appear in the tunnel of cervical cancer destiny. It is the change of the strategy from the fight against cervical cancer to the fight to reverse the Cervical Cancer Mortality curve in a definite population and for a definite time. The new strategy needs new tools and services. All of this is described in our book, What Every Women Should Know about Cervical cancer, Second Edition (Springer, 2017)

Parts were discussed at FASEB meetings in 2016 and 2017. This is a continuation of publishing the results of our research.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
How the DAG generation, transient shuttling of effector molecules and assembly of a signalosome at immunological synapse can regulate the early T cell response and L‐selectin shedding?
Authors: Aslam Naveed , Alvi Farah
Published: June 2021

Tunability of T cell activation profile could be key in orchestrating the optimal immune system response in pathologies like autoimmune, inflammation, allergies and some cancers. During early T cell activation events the fundamental proteolysis process of L‐selectin shedding is critically linked to the precision and specificity of immune response. Considerable evidence links the DAG homeostasis at immunological synapse to L‐selectin shedding and early T cell activation profile through DAG‐PKCα‐DGKζ‐Ras/ERK axis. Evidence also suggest that generation and consumption of this lipid at T cell‐APC contact zone must be exquisitely regulated due to its critical role in T cell polarization and activation. Here, we explore the question that how during the correct activation of antigen‐specific immune response DAG limits its own persistence? Understanding how the DAG homeostasis can be maintained could be significant for establishing a link between TCR triggering and Ras/ERK activation. Here, we report that a net negative feedback loop which is modulated by second messenger itself is responsible for maintaining the homeostasis and hence the precision and specificity of immune response. Here, we show that a local signalosome and functional sub‐signaling loop is formed between the components of DAG signaling which in turn, ensure the precision in degree of proteolysis of surface molecules such as L‐selectins. The coordinated translocation to I.S. in response to antigen recognition and subsequent activation of PKCα and DGKζ regulated the amplitude and duration of persistence of DAG levels. Our results propose that during the normal physiological conditions this sub‐signaling loop between DAG effector molecules is responsible for tightly maintaining the second messenger homeostasis however, under the pathological conditions this loop may become dysfunctional

Support or Funding Information

Private Funding

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cadmium‐induced Macrophage LDL and Oxidized LDL Internalization
Authors: Kumar Wasan Mani , Ribeiro Marta , Balla Andre
Published: June 2021

Environmental cadmium exposure has been established as a promotor of vascular inflammation and risk factor for atherosclerosis. Inflammation causes histone release, which was shown to increase macrophage low‐density lipoprotein (LDL) phagocytosis, which initiates foam cell formation, a crucial step in atherosclerosis. The link between histones and macrophages suggests cadmium may increase atherogenesis by upregulating scavenger receptors, which function to bind LDL and Oxidized LDL (OxLDL).

RAW 264.7 (murine) macrophages were incubated in a solution of DMEM (Dulbecco's Modified Eagle Medium) and treated with 2 μg LDL, 2 μg histones, and 10 nM, 50 nM, and 200 nM CdCl2 concentrations with varying incubation times (24 hr, 48 hr, and 72 hr). Cells were stained using Oil Red O (lipid sensitive dye) and imaged using a microplate reader at 500 nm. LDL was oxidized by incubation in 10 μM CuSO4 and centrifugal filtration in a PD‐10 spin column. OxLDL was diluted to 2 μg/ml and applied to macrophage cells with the aforementioned conditions, excluding histones. Oil Red O staining and absorbance measurement were performed with similar time constraints

In vivo studies for measurement of serum LOX‐1, CD‐36 and SR‐A I/II were also performed. LOX‐1, CD‐36, and SR‐A I/II are the main macrophage and endothelial receptors for oxidized LDL. ICR male Mice were exposed to cadmium through drinking water for 8 weeks. Mice were sacrificed, and serum was extracted for assessment of scavenger receptor expression by ELISA assay.

Oil Red O staining of RAW 264.7 cells incubated with 2 μg LDL, 2 μg histones and 10 nM, 50 nM, and 200 nM CdCl2 concentrations showed significant dose‐dependent and time‐dependent (24, 48, and 72 hr.) increases in lipid absorbance. Those at the highest CdCl2 concentrations showed the largest increase in LDL uptake, further augmented by histones. Similar results were found in macrophage cells incubated with 2 μg OxLDL. The serum ELISA provided inconclusive results to LOX‐1 serum concentrations.

Our data indicates cadmium modulates macrophage function to promote the uptake of LDL, histone‐LDL and OxLDL. We are currently studying the in vitro expression of major scavenger receptors, including LOX‐1, CD‐36 and SR‐A I/II that may provide insight into the mechanism of action for cadmium adverse effects on macrophages.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dose‐specific morphological changes in the Sertoli cell of the adult male Japanese quails testes exposed to di(n‐butyl)phthalate DBP prepubertally
Authors: Bello Umar M. , Aire Tom A. , Imam Jibrin , Abdulazeez Jimoh , Igbokwe Casmir O.
Published: June 2021

Di(n‐butyl) phthalates (DBP) are endocrine‐disrupting chemicals (EDCs) implicated in a number of reproductive disorders in humans and rodents. Previously, we have shown that DBP effects is anti‐androgenic and causing alterations in seminiferous tubular histology and impairment of Leydig cell steroidogenesis in the avian model. However, the ultrastructural changes on the Sertoli cell remain unclear. The present study was designed to assess, morphologically, the dose‐dependent toxic effect of pre‐pubertal exposure of the plasticizer‐DBP, on the histology and ultrastructure of the Sertoli cell. DBP was administered to 10‐week old male quail birds by oral gavage, with a control group given corn oil vehicle(1mL/kg) only, while the other five experimental groups were fed (1, 10, 50, 200– and 400mg/kgbwt/d DBP(dissolved in corn oil) , for a period of 30days. Testes were fixed in Bouin's liquid or in a mixture of 2.5% glutaraldehyde and 2% formaldehyde for analysis under light and electron microscopes(TEM), respectively. Histologically, the basal Sertoli cell cytoplasm in high DBP dosage levels (200mg and 400mg/kgbwt/d), increasingly displayed aggregations of lipid droplets, but only a few in the apical Sertoli cell cytoplasm. In addition, there was marked vacuolations in the lining germinal epithelium. Ultrastructurally, there were notable, dose‐dependent changes seen in the Sertoli cell in DBP‐treated birds, with low dosage groups displaying no obvious structural changes, while the DBP high‐dose resulted in a remarkable accumulation of numerous, scattered or usually, closely packed lipid droplets, found predominantly in the basal part of the Sertoli cell cytoplasm. In addition, there were several mis‐shaped or swollen mitochondria with electron lucent areas, indicative of early cellular degeneration. These degenerating mitochondria were characterized by swelling and deformation of the tubular cristae, resulting in a “moth‐eaten” appearance. These findings showed that high dose of DBP (200 and 400mg/kgbwt/d) administered daily for 30days significantly resulted in degenerative characteristics, such as the increase in lipid droplets, intra‐epithelial vacuolations and mitochondrial damage in the Sertoli cell. Thus, this study has provided a clear evidence that alterations in Sertoli cell morphology and function were involved in spermatogenic failure induced by DBP, in this specie.

Support or Funding Information

The research was supported by Ahmadu Bello University Board of Research and grants from the South African Veterinary/Novartis Wildlife Foundation.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Repurposing neuroleptic drug for brain tumor therapy
Authors: Kaushik Itishree , Srivastava Sanjay
Published: June 2021

Brain tumor is considered as one of the most aggressive and incurable form of cancer. Majority of the brain tumors have a median survival rate of only 12%. Even though advanced treatments such as surgical removal, chemotherapy and radiotherapy are available, brain tumor persists to be lethal. Temozolomide (TMZ) is the main therapy for brain tumor but resistance to TMZ is the major cause of treatment failure. This resistance in tumor cells has frequently been linked to the expression of O6‐methylguanine‐DNA methyltransferase (MGMT). Pimozide (PMZ) is a neuroleptic drug used for the treatment of schizophrenia and chronic psychosis. In our study, we observed that pimozide significantly reduced the viability of U87MG, U251MG, DAOY and GBM28 cell lines with an IC50 ranging from 11μM to 20μM after 24 h of treatment. Pimozide induced apoptosis in these cell lines as evaluated by FITC/Annexin assay and further validated by the cleavage of caspase 3 as well as PARP. Western blot analysis resulted in the concentration dependent decrease in the expression of STAT3 signaling mainly targeting the anti‐apoptotic markers BCl‐XL, BCl‐2 and Survivin etc in U87MG, U251MG, DAOY and GBM28 brain cancer cell lines. PMZ alone suppressed the growth of U87MG cells with an IC50 of 10μM, after 96 hours of treatment. On the other hand, treatment of U87MG cells with 750 μM of TMZ in combination with 10μM PMZ for 96 hours increased the killing of U87MG cells by 50% as compared to 10% when treated with TMZ. In addition, PMZ treatment reduced the survival of TMZ resistant U251R cells with an IC50 ranging from 15–20 μM after 24 h of treatment. Our results further demonstrated that oral administration of PMZ (25mg/kg) significantly inhibits the growth of brain tumors in an intracranial tumor model without any apparent signs of toxicity. It is important to note that PMZ is an FDA approved drug with no considerable toxicity. Overall, this study depicts that PMZ is a potential candidate to target brain tumors.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Case‐Based Discussion to Review Infectious Disease Therapeutics in a Graduate Physician Assistant Studies Program
Authors: Guillory Ashley Nicole
Published: June 2021

Infectious disease therapeutics have historically been a difficult topic for the physician assistant students at University of Texas Medical Branch. Average test scores on the exam covering these topics are typically 5–10 points below the class average of other exams. In order to improve students' test scores and understanding of this material, a case‐based review session was added at the end of the infectious disease module.

The review session began with a short lecture (~30 minutes) reviewing high yield concepts as preparatory material. The lecture slides were made available to students 2 days prior to the review session. Students were divided into small groups and assigned 1–2 short cases covering the different classes of infectious disease therapeutics. Each case included questions related to choosing the most appropriate treatment, the pharmacological characteristics of the treatment, and alternatives due to patient characteristics such as renal failure, pregnancy, and hepatic insufficiency. Students worked through cases in their small groups then reconvened to present their case and answers to the entire class. A post‐session survey was given to determine both student satisfaction with the session as well as the impact of the session on their knowledge and understanding of the material. Effectiveness was assessed by the test scores on the exam for the infectious disease module.

Prior to the review session, 82% of survey respondents indicated that they lacked confidence in their knowledge of infectious disease topics. Following the review session, only 23% of respondents indicated that they still lacked confidence in their knowledge of these topics. 97% of survey respondents indicated that the review session was beneficial with the majority of respondents indicating that it was very beneficial. The survey also included a free response question for students to offer feedback on the session and ways to improve the session. Themes that were discovered in these answers included: 1) students felt that they should have studied more before attending the session, 2) have the cases instructor led rather than the majority of the discussion happen in the individual groups, and 3) incorporate the case studies into the lecture portion. Overall, the response to the review session was largely positive and the feedback will be useful for improving the review session next year. This format will be useful in developing review sessions for other pharmacology topics that the students struggle with.

Support or Funding Information

This study was not supported by a grant.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
“Memory clinics:” A complement to teaching and learning in pharmacology
Authors: Silva Ricardo Alfonso Pena , Larrarte Maria Alejandra
Published: June 2021

Pharmacology is considered one of the most difficult subjects by medical students. One of the reasons that explains the difficulty is the need to learn multiple names of medications that are not familiar to the medical student. To support the learning of pharmacology, we have developed "memory clinics" to offer a metacognitive space that allows the student to reflect on their study methods and fears when learning pharmacology.

"Memory clinics" are a voluntary activity in which medical pharmacology students can participate. The activity takes place in several stages during a one‐hour block. 1) Two short study sessions (individually, individually or in groups) to memorize biomedical information related to pharmacology. 2) retention of information is evaluated in a short activity in which the students solve a crossword puzzle. At the beginning, in‐between the sub‐stages and at the end of the activity the students answer some open questions in which they reflect about confidence in their mastery of the concepts and time devoted to study and answer questions. At the end of the activity, the correct answers to the questions are given and each student evaluates his/her own performance.

The piloting of innovation shows that students are very pleased with the activity and recognize the importance of having a space to reflect on their learning processes. At the beginning, only three "memory clinics" were planned, but students have requested that more activities of this type be included, because they feel that innovation has favored their learning process.

Support or Funding Information

College of Medicine, Universidad de los Andes

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Clinical Pharmacology Learning Tools for the Tech Savvy in a Physician Assistant (MPA) Program
Authors: Ram Arun M , Rajasekaran Senthilkumar
Published: June 2021

Students of our MPA program are expected to master a vast amount of information in a short 28‐ month curriculum leading up to their clinical practice. In addition, students find learning clinical pharmacology difficult as it involves memorizing hundreds if not thousands of medications, their mechanism, interactions, and adverse effects. In order to maximize learning, we delivered a Clinical Pharmacology (CP) course in a flipped classroom model using technology1.

CP I and II are two 3‐credit courses taught over 4‐month each with two 2‐hour didactic lectures every week. In its place, we introduced short 15 – 20 minute videos recorded using Explain Everything or Camtasia software and posted them a couple of days prior to in‐class sessions2. In addition, a complete slide‐deck and a study guide with open‐ended questions designed to reinforce the clinically relevant facts and concepts in each class were also posted. Students listened to the recording, used the slide‐deck and study guide to master the concepts. An anonymous survey at the end of the course collected the feedback on this new model of course content delivery and their learning experience3.

More than 55 students participated in the survey. An overwhelming 82% of the respondents wanted the course to be delivered based on this model in the future. About 75% agreed that the new model allowed them more time to study other subjects. Another 76% felt the new model helped them feel prepared for the exams. Although we had reduced the student‐faculty contact time by 50%, about 70% of the survey respondents felt the new model provided them adequate faculty exposure and interaction.

Unlike reported by some studies that the students prefer only about half of the class sessions to be “flipped” 4; our MPA students wanted us to follow this new model for the entire course. Following our success with this new model, other Course Directors have started introducing flipped classes in their courses.

Based on the favorable feedback we received from the students and performance index on their course assessments, we recommend utilizing this model of active learning in your health sciences education programs.

Support or Funding Information

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.