bluuebunny/crossref_metadata_2025_split huggingface dataset:
When Wolff's law isn't enough: training students to consider soft tissues in promoting skeletal health
Authors: Hamrick Mark 
Published: June 2021

The physiology of various organ systems has revealed extensive crosstalk among tissues such as adipose tissue, skeletal muscle, and bone. These findings have important implications for skeletal biology, particularly within the context of improving bone health during growth and with aging. The latter is especially significant for students in the health sciences, since the population over the age of 65 is the most rapidly growing demographic globally and in the United States. Mechanical loads emanating from body weight and muscle contraction are important for bone mass accrual early in life, but the response of bone to mechanical stimuli is markedly attenuated in adults. Though increased loading magnitude and frequency may have less dramatic anabolic effects on the adult skeleton than on young bones, inactivity and muscle disuse atrophy can contribute significantly to age‐related bone loss. This point is reinforced by data from clinical studies showing that older adults with low appendicular lean mass and reduced grip strength also have reduced cortical bone area and thickness. The highly integrated changes in skeletal muscle and bone observed in aging patient populations suggest that targeting either sarcopenia or osteoporosis alone may not be sufficient for effective fracture prevention. Thus, new strategies for improving both tissues simultaneously to prevent “osteosarcopenia” have been proposed. These recent advances underscore the importance of considering soft‐tissues for maintaining bone health and preventing falls and bone fractures.Support or Funding InformationNational Institute on Aging (AG 036675)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 Rationally Engineered Hyperactive Actin‐Resistant DNase1‐Fc Fusion Protein Ameliorates Autoimmune Glomerulonephritis
Authors: Austin Cary D. , Mouchess Maria , Suto Eric , Chan Sara , Dai Bingbing , Santell Lydia , Tam Christine , Hackney Jason A. , Cao Yi , Luan Peng , Darwish Martine , He Dongping , Haller Susan , Eastham‐Anderson Jefferey , Franke Yvonne , Lu Yanmei , Blanchette Craig , Lee Wyne P. , Lazarus Robert A. , Yi Tangsheng 
Published: June 2021

Loss of B cell tolerance and generation of autoreactive anti‐nuclear antibodies are hallmarks of systemic lupus erythematosus (SLE) and lupus nephritis. Lupus nephritis is characterized by glomerular and tubulointerstitial inflammation often initiated by the renal glomerular deposition of anti‐nuclear immune complexes which trigger subsequent activation of complement, macrophages/monocytes and other innate inflammatory cells. The mechanism of anti‐nuclear immunoglobulin accumulation and clearance in lupus nephritis pathogenesis remains largely uncharacterized. Here, we show that innate immune activation in the NZB/W F1 mouse model and in human lupus nephritis biopsies rapidly reduces DNase1 expression in renal cortex proximal tubular cells. To overcome the loss of endogenous DNase1, we treated lupus‐prone mice with a hyperactive actin resistant variant of DNase1 with improved catalytic activity against nucleic acid‐IgG immune complexes and acceptable in vivo pharmacokinetics. Hyperactive DNase1‐Fc fusion protein ameliorates nephritis in a murine model of lupus nephritis and reduces immune complex deposition/complement fixation. Taken together, our data suggest that the loss of renal DNase1 through TLR signaling or other innate immune activation impairs clearance of autoreactive anti‐nuclear immune complex deposits in the kidney to promote nephritis progression. Our findings provide a therapeutic rationale for using an engineered DNase1‐Fc as a potential therapeutic approach in lupus nephritis.Support or Funding InformationAll authors are current or previous employees of Genentech, Inc.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of gene augmentation strategy for the treatment of MAK associated Retinitis Pigmentosa
Authors: Gibson‐Corley Katherine N. , Cranston Cathryn , Anfinson Kristin , Kaalberg Emily , Han Ian , Slusarski Diane C. , Goeken Adam , Businga Thomas , Brown Hannah , Mullins Robert F. , Stone Edwin M. , Tucker Budd A. 
Published: June 2021

PurposeThe gene male germ cell associated kinase (MAK) encodes a protein involved in regulation of photoreceptor cilia length. By combining next generation whole exome sequencing and induced pluripotent stem cell (iPSC) technology we found that an Alu repeat inserted in exon 9 of the MAK gene resulted in a loss of normal MAK transcript and development of human autosomal recessive retinitis pigmentosa (RP). Although a relatively rare cause of disease in the general population, the MAK variant is enriched in individuals of Ashkenazi Jewish descent. In this population, 1 in 55 individuals are carriers and 1/3 of all cases of recessive RP is caused by this gene. The purpose of this study was to determine if a viral gene augmentation strategy could be used to restore functional MAK protein and in turn would be a good candidate for treatment of early stage MAK associated RP.MethodsTo demonstrate the ability of the developed MAK gene transfer vector to restore normal transcript and protein, patient specific iPSC derived photoreceptor precursor cells were transduced and immunocytochemistry, rt‐PCR and western blot analysis were performed. To demonstrate transgene function, cilia length assays were performed using patient derived fibroblast cells in vitro and MAK knock out zebra fish in vivo. In addition, visual function testing was performed in the zebra fish. To demonstrate safety of cGMP generated clinical grade vector, local and systemic toxicity studies were performed via subretinal injection into wildtype rats.ResultsMAK mutant RP iPSC derived photoreceptor cells harboring the previously identified ALU insertion (Tucker et. al. 2011) were generated and transduced with viral vectors containing the retinal MAK transcript. At 1‐week post‐transduction normal retinal MAK transcript and protein could be detected via rt‐PCR and western blotting respectively. Using patient derived fibroblast cells and MAK knockdown zebra fish we demonstrate that over expression of retinal MAK transgene restored primary cilia length. In addition, the visual defect identified in MAK knockdown zebra fish was mitigated via treatment with retinal MAK transgene. Following subretinal delivery of clinical grade vector into wild type rats no evidence of local or systemic toxicity was detected at either 1 or 3 months' post‐treatment.ConclusionsWe have successfully developed a MAK gene replacement strategy that has been validated in human iPSC derived photoreceptor precursor cells in vitro and MAK knock down zebra fish and wildtype rats in vivo.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of CaS nanostructures in growth and proliferation of melanoma and benign skin cells in vitro
Authors: Pérez‐Caro Cristina Ivelisse , Narváez‐Ramos Michelle Angelie , Rodríguez‐Martínez Olga , Muñoz‐Muñoz Keyshla , Suarez‐Martínez Edu B , Diffoot‐Carlo Nanette , Castro‐Rosario Miguel 
Published: June 2021

One person dies of melanoma skin cancer every hour according to the American Cancer Society. The Skin Cancer Foundation highlights that melanoma is the most dangerous form of skin cancer. The standard treatment involves the surgical removal of the tumor with follow up treatments depending on the cancer stage. Once skin cancer melanoma metastasizes, it is very difficult to treat. We have extended our earlier work on the use of calcium sulfide nanostructures to skin cancer melanoma in vitro. We studied the effect of CaS nanostructures in the growth and proliferation of the melanoma ATCC CRL‐7637 skin cancer cells as well as the corresponding pre‐control ATCC CRL‐7636 benign cells and control ATCC CRL‐2106. The results were consistent with our previous studies on mammary adenocarcinoma and lung cancer cells where the malign cell growth and proliferation were limited by the addition of CaS nanostructures with no effect on the growth and proliferation of benign cells over a period of 0, 24, 48 and 72 hours. Work in progress addresses the effect of these nanostructures in the cell life cycle.Support or Funding InformationFinancial support from the NIH RISE‐E‐BASE under grant number 1 R25 GM127191‐01 is gratefully acknowledged.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparison of Different Methodologies and Cryostat versus Paraffin Sections for Chromogenic Immunohistochemistry
Authors: Hira Vashendriya , de Jong Annique Loncq , Ferro Klea , Khurshed Mohammed , Molenaar Remco , Van Noorden Cornelis J.F. 
Published: June 2021

Immunohistochemistry (IHC) specifically localizes proteins in cells and tissues, but methodologies vary widely. Therefore, we performed a methodological IHC optimization and validation study. First, we compared advantages and disadvantages of cryostat sections versus paraffin sections. Second, we compared and optimized antigen retrieval in paraffin sections using citrate buffer and Tris/EDTA buffer. Third, aminoethyl carbazole (AEC) and 3,3′‐diaminobenzidine (DAB) were tested as horseradish peroxidase (HRP) substrates to obtain a water‐insoluble coloured end product to visualize antigens. Fourth, secondary antibodies conjugated with either mono‐HRP or poly‐HRP were compared. The study was performed using serial sections of human tonsil. IHC was performed with primary antibodies against endothelial cell marker CD31, smooth muscle actin (SMA), chemokine stromal‐derived factor‐1α (SDF‐1α) and its receptor C‐X‐C receptor type 4 (CXCR4), macrophage marker CD68 and proliferation marker Ki67. DAB rather than AEC, and cryostat sections rather than paraffin sections gave optimum staining at highest primary antibody dilutions, whereas tissue morphology in paraffin sections was superior. Loss of antigenicity in paraffin sections by formaldehyde fixation, heat and/or masking of epitopes was counteracted by antigen retrieval but not for all antigens. Two out of six antigens (CD31 and CD68) could not be retrieved irrespective time and type of retrieval. Tris‐EDTA was superior to citrate buffer for antigen retrieval. The use of mono‐HRP or poly‐HRP depended on the affinity of the primary antibody for its antigen. We conclude that IHC methodology optimization and validation are crucial steps for each antibody and each research question.Support or Funding InformationKWF; UVA 2014‐6839This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Systemic Inflammatory Activation of Astrocytes in Postnatal Development
Authors: Alves Michele Joana , Blackbourn Jessica , Melo e Silva Talita , Alzate Diego , Jones Mikayla , Otero Jose , Czeiler Catherine 
Published: June 2021

Systemic inflammation during the postnatal period triggers neuroinflammation which can have a multitude of deleterious effects on the newborn, including increased susceptibility to hypoxia. Astrocytes in the brainstem are capable of sensing oxygen levels and modulating the respiratory response accordingly. Astrocytes are also crucial players in neuroinflammation by regulating immune responses in the central nervous system. Upon activation, astrocytes promote the release of cytokines, which trigger immune cell infiltration and contribute to the exacerbation of existing inflammation. Paradoxically, astrocytes can also exert neuroprotective effects during activation. Therefore, it is important to understand the mechanism whereby astrocytes become activated and the factors affecting astrocyte behavior during inflammation. One of the hallmarks of astrocyte activation is a change in morphology and an increase in migratory behavior. It is unclear how this global change in morphology and migratory behavior affects the astrocyte's ability to sense and react to changing oxygen levels. In this study, we investigate the effects of systemic inflammation on the morphological/migratory pattern and oxygen sensing capacity of astrocyte populations derived from the hindbrain of 5‐day‐old mice. Results of this study increase our current understanding of astrocyte functional and morphological responses to an inflammatory milieu.Support or Funding InformationThis work was supported by NIH grant R01HL132355.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Measurement of sedation in individual Drosophila
Authors: Clowes Katherine Rose , Petersen E. Nicholas , Hansen Scott 
Published: June 2021

Though widely used, the mechanisms of sedation from compounds like ethanol and volatile anesthetics are not well understood. Drosophila melanogasterhas been identified as an ideal model organism for studying phenotypic responses to ethanol and other anesthetics due to its low cost, similar phenotypic response, and many available genetic tools. Previously, measuring the effects of anesthetics on Drosophila has required labor‐intensive methods often using large cohorts of flies, making them cumbersome and inefficient. Here, we describe the setup and experimental protocol for Volatile Anesthetic Position Recording (VAPR), a high‐throughput technique to measure the effect of ethanol vapor and other volatile drugs on Drosophila behavior. VAPR utilizes flow meters to adjust the relative concentration of aerosolized drugs which are then passed through chambers containing individual flies. Automated video tracking records the position and movement of the flies over time, which we analyze to quantify hyperactivity, sedation, and other treatment related behavioral changes. Using VAPR, we found flies lacking the enzyme phospholipase D (PLD) are insensitive to ethanol sedation but appear unaffected in their sensitivity to the anesthetics chloroform and isoflurane.Support or Funding InformationNational Science Foundation (NSF) Summer Undergraduate Research Fellowship (SURF) grant 1359369.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
APP21 transgenic rats develop age‐dependent cognitive impairment and microglia accumulation within white matter tracts
Authors: Liu Qingfan , Weishaupt Nina , Shin Sheojung , Singh Ramandeep , Agca Yuksel , Agca Cansu , Hachinski Vladimir , Whitehead Shawn 
Published: June 2021

BackgroundMost of the animal models commonly used for preclinical research into Alzheimer's disease (AD) largely fail to address the pathophysiology, including the impact of known risk factors, of the widely diagnosed sporadic form of the disease. Here, we use a transgenic rat (APP21) that does not develop AD‐like pathology spontaneously with age, but does develop pathology following vascular stress. To further the potential of this novel rat model as a much‐needed pre‐clinical animal model of sporadic AD, we characterize APP21 transgenic rats behaviorally and histologically up to 19 months of age.MethodsThe open field test was used as a measure of activity; and the Morris water maze was used to assess learning, memory, and strategy shift. Neuronal loss and microglia activation were also assessed throughout the brain.ResultsAPP21 transgenic rats showed deficits in working memory from an early age, yet memory recall performance after 24 and 72 h was equal to that of wildtype rats and did not deteriorate with age. A deficit in strategy shift was observed at 19 months of age in APP21 transgenic rats compared to Fischer wildtype rats. Histologically, APP21 transgenic rats demonstrated accelerated white matter inflammation compared to wildtype rats, but interestingly no differences in neuron loss were observed.ConclusionsThe combined presence of white matter pathology and executive function deficits mirrored what is often found in patients with mild cognitive impairment or early dementia, and suggests that this rat model will be useful for translationally meaningful studies into the development and prevention of sporadic AD. The presence of widespread white matter inflammation as the only observed pathological correlate for cognitive deficits raises new questions as to the role of neuroinflammation in cognitive decline.Support or Funding InformationWe appreciate funding support from the Ontario Mental Health Foundation, and funding from the Canadian Institutes for Health Research, Canadian Consortium on Neurodegeneration in Aging and Canadian Foundation of Innovation.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Research on the standardized training mode of clinical medical talents based on clinical pathway
Authors: Duan Hubin , Hao Chunyan , Wang Suping , Zhang Xuefeng 
Published: June 2021

ObjectiveTo study the effect of clinical pathway on the ability of standardized treatment of patients for medical students.Methods450 undergraduates were divided into group A and group B, Group A was the traditional practice group, group B was the clinical pathway teaching group. 225 students in each group. group A students according to the traditional practice teaching method, finished the ward round and case discussion with their teachers, consulted the attending physician's advice for their patients. On that basis, tried to give the simulation of the doctor's advice after choosing the same disease but different patients, recording their history, according to their own understanding; Group B students studied the clinical pathway in advance, based on the method of Group A, followed the clinical pathway, combined with their own understanding, tried to give the simulation of the doctor's advice. Two high qualificated experts inspected the completeness and correctness of the simulation of the doctor's advice giving by group A and group B students. At the same time , students' confidence in independent treatment of same disease were assessed by themselves.Resultsthe integrity of students' medical advice in group A and group B was 85.68±3.10 vs 92.05±2.96‐P<0.01; The correctness was 86.55±3.88 vs 95.59±4.31‐P<0.01; The degree of confidence in the treatment of the doctor's order was 75.19±6.90 vs 90.45±5.65‐P<0.01.ConclusionThe students in the clinical pathway‐teaching group are better than the traditional teaching group students in the confidence, the integrity and the correctness of the doctor's orders.Support or Funding InformationThis project is supported by the teaching reform program of shanxi higher education (No. 2013‐ 033), The project of "131" leading talents of shanxi higher education, the education reform project of postgraduate students in Shanxi Province (No. 2017‐027), the education reform project of shanxi medical university, the National Natural Science Foundation Youth Fund (30600637). China Postdoctral Foundation(2014M561207)‐Shanxi Scholarship Council of ChinaThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ozone‐Induced Systemic Metabolic Alterations are Regulated by Circulating Stress Hormones in Rats
Authors: Snow Samantha J , Henriquez Andres R , House John S , Schladweiler Mette C , Miller Colette N , Kodavanti Urmila P 
Published: June 2021

Several recent studies from our lab and others have demonstrated widespread metabolic effects following an acute ozone exposure. Although these effects are similar to those induced by the fight‐or‐flight response involving adrenal‐derived stress hormones, little data are available linking these metabolic changes to activation of neuroendocrine pathways. In this study, we used surgical and pharmacological interventions to understand the influence of adrenal‐derived stress hormones in mediating ozone‐induced metabolic alterations. Male Wistar‐Kyoto (WKY) rats underwent bilateral total adrenalectomy (ADREX) or sham surgery prior to their exposure to filtered air or ozone (0.8 ppm), 4h/day for 1 day. In the sham animals, ozone exposure induced hyperglycemia and glucose intolerance as well as increased levels of circulating total cholesterol, free fatty acids (FFA), and leptin. ADREX and the resultant depletion of adrenal‐derived stress hormones, adrenaline and corticosterone, significantly attenuated these ozone‐induced metabolic effects. Furthermore, ozone‐induced global transcriptional changes in the liver from sham and ADREX rats were assessed by total mRNA sequencing. The upstream regulator assessment within Ingenuity Pathway Analysis revealed that ozone‐induced changes in gene expression were similar to those induced by corticosterone, methylprednisolone, and forskolin, suggesting glucocorticoids and adrenergic mechanisms significantly contributed to the ozone‐induced changes in the liver. These same regulators were not predicted in the ADREX animals exposed to ozone. In a follow up study, we utilized a non‐specific beta‐adrenergic receptor antagonist (propranolol) and/or glucocorticoid receptor antagonist (mifepristone) to understand the specific contribution of these stress hormones in mediating metabolic effects following ozone exposure. Ozone‐induced hyperglycemia, glucose intolerance, and increased FFA levels were significantly attenuated only by the combination of antagonists, suggesting that both adrenergic and glucocorticoid pathways were involved in the regulation of these processes. However, increases in total cholesterol and leptin levels following ozone exposure were blocked by mifepristone alone and in combination with propranolol, suggesting these ozone effects are glucocorticoid‐dependent. Collectively, these data suggest adrenal‐derived stress hormones play a significant role in mediating ozone‐induced systemic metabolic effects. (Does not reflect the US EPA policy)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.