bluuebunny/crossref_metadata_2025 huggingface dataset:
The effectiveness of flaxseed (Linum usitatissimum L.) on the inflammatory response in patients with familial hypercholesterolemia receiving lipid apheresis‐preliminary results
Authors: Kanikowska Dominika , Rutkowski Rafał , Pawlaczyk Krzysztof , Sato Maki , Misian Monika , Bręborowicz Andrzej , Witowski Janusz
Published: June 2021

Familial hypercholesterolemia is associated with severe abnormalities including atherosclerosis, dyslipidemia, cardiovascular disease, oxidative stress, inflammation, and endothelium dysfunction. Flaxseed in the diet acts upon blood lipids, lowering cholesterol, and also has anti‐oxidative effects. These effects are beneficial for patients suffering from atherosclerosis, dyslipidemia and endothelium dysfunction.

The protocol consists of four stages: a run‐in phase of 10 weeks (patients on a standard diet) and an experimental stage ‐ flaxseed supplementation or placebo (whole‐wheat) supplementation, and wash out phase of 10 weeks. Samples of saliva and blood were collected after each phase of the trial, just before routine lipoprotein apheresis, which is usually once every two weeks. Saliva myeloperoxidase levels were analyzed by ELISA test, and flow cytometry was used for measurement of circulating microparticles. The study adhered to the Principles of the Declaration of Helsinki.

The concentration of salivary myeloperoxidase was lower after flaxseed supplementation.

The number of circulating microparticles: CD105 (endothelial‐derived), CD45 and CD41 (platelet‐derived)) was smaller after flaxseed supplementation compared with a run in phase

Conclusions

Our preliminary observations indicate that a diet containing flaxseed may improve endothelial cell function and thus contribute to the amelioration of unfavorable effects associated with hypercholesterolemia.

Support or Funding Information

There is no actual or potential conflict of interest in this presentation. The Nutricia Research Foundation Poland provided the research grant support.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Predicting Protein Glycation Rate and State: The Need for Models to Incorporate Additional Features
Authors: Turkette Thomas , Rhinesmith Tyler , Root‐Bernstein Robert
Published: June 2021

Glucotoxicity is considered to be one of the primary drivers in type 2 diabetic (T2D) pathophysiology. As such, understanding the mechanisms underlying glucotoxicity is imperative in the identification, treatment, and prevention of T2D. One contributor to glucotoxicity is protein glycation in which a reducing sugar spontaneously reacts with the ɛ‐amino group of lysine residues or an N‐terminal amino group to form a covalently bound adduct. Here we test two methods employed to predict whether a given protein sequence will undergo glycation and whether either method offers insight toward predicting the rate of glycation in vitro. The first method involves looking for sequences that are homologous to known glucose binding regions, while the second utilizes NetGlycate‐ a neural network trained to predict glycation based on a protein sequence. Both methods were used to predict potential glycation sites on the human insulin receptor and regions containing these sites were synthesized. These fragments were then incubated in high glucose for 1, 3, or 6 days. The extent of glycation was assessed using MALDI‐ToF mass spectrometry. There was some overlap in the residues predicted to undergo glycation, but both methods also predicted unique sites. Both methods generated a number of false positives, but sites predicted by both methods were generally more likely to undergo glycation. Additionally, several of the sites predicted that were unique to one method underwent glycation. The rate of glycation for a given site varies greatly regardless of the method employed to predict it and this holds true even when both methods predicted the same site. Our results suggest: 1) neither method is sufficient to independently predict protein glycation consistently and accurately, 2) a model that incorporates both evolutionary considerations in addition to protein sequence would be expected to better predict protein glycation, and 3) neither approach informs the rate of protein glycation, suggesting that some other factor(s) must also be at play.

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 New Player in Intra‐islet Communication: The Hyperglycemic Effects of Neuronostatin through Increased Glucagon Production and Reduced Insulin Release
Authors: Grote Stephen , Yosten Gina , Samson Willis
Published: June 2021

The tight regulation of blood glucose with insulin and antidiabetic agents greatly increases the risk and dangers of hypoglycemia in patients with Type 1 and Type 2 Diabetes. In addition, repeated bouts of hypoglycemia diminish the sympathetic arms of the counterregulatory response and blunt awareness of symptoms, creating a vicious cycle of unawareness and increasingly severe hypoglycemia. The only option in preventing or reversing hypoglycemia unawareness is careful avoidance of hypoglycemia through close monitoring– an often unobtainable goal. There is great need for other therapies and better understanding of diabetic hypoglycemia. We recently identified a novel peptide, neuronostatin (NST), encoded in the somatostatin preprohormone and found in all somatostatin‐positive tissues and cells. However, the activity of NST is functionally distinct from other preprosomatostatin products. Furthermore, it appears the physiologic stimuli of the peptides are different and that the peptides are sequestered in separate secretory vesicles. Our group identified the orphan GPCR, GPR107, as a cognate receptor of NST. Through an interaction with GPR107, NST enhanced proglucagon mRNA expression and glucagon release from isolated islets and immortalized alpha cells. NST also reduced beta cell release of insulin in response to high glucose conditions in isolated islets. Based off our findings, we hypothesized that NST is a pancreatic component of the counterregulatory response to hypoglycemia. In support of our hypothesis, an infusion of NST substantially increased plasma glucose levels over a 30‐minute time period in male rats. Additionally, NST delayed glucose clearance and diminished the insulin response to hyperglycemia in a glucose tolerance test. Fasting blood glucose raised plasma NST levels in rats, and exposure to low glucose stimulated NST release from human islets. Further experiments will confirm the role of NST in preventing or reversing hypoglycemia and determine how NST plays a role in the complex relationships of islet regulation and signaling. We propose that NST could represent a novel therapeutic target for the treatment and prevention of hypoglycemia in diabetes.

Support or Funding Information

This work was completed with funding support from the NIH and NIDDK, and the Helmsley Charitable Trust.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Constructing glucagon biosensors as an alternative to conventional detection methods
Authors: Ennis Teresa Annmarie , Samson Willis K. , Yosten Gina L. C.
Published: June 2021

Glucagon is a counterregulatory hormone, produced by the pancreatic alpha cells, that is essential for protection against hypoglycemia. In patients with Type 1 Diabetes, failure of appropriate glucagon responses can lead to severe hypoglycemia, coma, and death. Studies into the regulation of glucagon secretion have been hindered by a lack of assays that exhibit adequate specificity and sensitivity. To overcome this limitation, we constructed glucagon biosensors expressing the glucagon receptor and the calcium sensor, GCamP6s. First, HEK‐293 cells were transfected with a GCGR (RFP‐Tagged) Human Glucagon Receptor vector, followed by transfection with the GCamP6s calcium reporter (GFP). After transfection, the cells were then passaged and maintained with media containing puromycin and G418 to isolate the cells that took up both vectors. Expression of the vectors could then be detected and monitored for changes stimulated by glucagon under different glucose conditions. After applying varied concentrations of glucagon, the GFP signal was detected using microscopy. The images produced clearly showed the receptor was visible in the membrane, and when glucagon was present the GCamP6s sensor was activated. In future studies, these glucagon‐sensing cells can be utilized to detect and monitor glucagon release from isolated islets, offering an alternative to conventional detection methods such as ELISA.

Support or Funding Information

NIH

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lactobacilli ‐induced Generation of Reactive Oxygen Species via Formyl Peptide Receptor‐1 (FPR1) Regulates Intestinal Motility in Mice
Authors: Chandrasekharan Bindu , Saeedi Bejan , Alam M Ashfaqul , Srinivasan Shanthi , Tansey Malu , Jones Rheinallt , Nusrat Asma , Neish Andrew S
Published: June 2021

Rationale

Various clinical conditions like diabetes and inflammatory diseases of the gut are associated with gastrointestinal (GI) motility disorders. Despite the increased consumption of beneficial commensal microbes marketed as ‘probiotics’, their mechanism of action on the enteric nervous system (ENS) is still unclear.

Aim

We studied the effects of the widely used probiotic and normal commensal Lactobacillus rhamnosus GG (LGG) on ENS and GI motility.

Methods

Conventional (Conv), formyl peptide receptor‐1 (FPR1 KO) and formyl peptide receptor‐2 (FPR2 KO) knockout mice were gavaged with hanks buffered salt solution (HBSS, negative control) or LGG. Mice were sacrificed after 2 h, and the jejunum was cryofixed and immunostained for phospho‐Erk (p44/42 MAPK, mitogen activated protein kinase). In a separate experiment, mice were intra peritoneally injected with a fluorescent hydro‐cy3 dye prior to HBSS/LGG gavage, and production of reactive oxygen species (ROS) was assessed in the longitudinal muscle myenteric plexus (LMMP) by confocal imaging after 2h. Ileum was assayed for gene expression of neuropeptides and Hand‐2 (transcription factor promoting enteric neuronal differentiation) by real time polymerase chain reaction (qRT‐PCR). Fluorescence in situ hybridization (FISH) was done on ileal cryosections from conventional mice using FPR1 and peripherin (pan neuronal marker) RNA probes. Conv mice were gavaged daily with HBSS or LGG for 2 weeks, and motility was assessed from stool frequency, total gastrointestinal transit time and ex vivo studies on isolated circular muscle strips by isometric muscle recording.

Results

LGG stimulated myenteric ROS production, enhanced Erk 1/2 phosphorylation and upregulated choline acetyl transferase (ChAT) neurons (P < 0.001) in Conv and FPR2 KO mice. These effects were abrogated in FPR1 KO mice, suggesting that LGG‐mediated signaling in the ENS is redox‐dependent and requires FPR1(P < 0.001), localized on enteric neurons by FISH assay. Functionally, LGG gavage for 2 weeks significantly improved stool frequency, reduced total GI transit time and enhanced ileal circular muscle strip contractions (P < 0.05).

Conclusions

Our study demonstrates for the first time, the presence of FPR1 on enteric neurons, and the LGG‐FPR1‐dependent redox‐signaling pathway that could be exploited to improve GI motility.

Support or Funding Information

We acknowledge support from the U.S. National Institutes of Health grant AI64462 (A.S.N.) and DK089763 (A.N. and A.S.N.)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Chrysin on intestinal inflammation and gut microbiota in lipopolysaccharides‐induced mice
Authors: Wu Wen‐Tzu , Wang Zhi‐Hong
Published: June 2021

Chrysin is a natural flavonoid with a variety of biological activities such as antioxidant, antivirus, antitumor, antidiabetic and so on. However, the potential protective effects of chrysin, against lipopolysaccharides (LPS)‐induced intestinal inflammation and its associated alteration of gut microbiota have not been studied. In the present study, male C57BL6/J mice were randomly divided into three groups and fed either normal chow diet (CON and LPS groups) or that supplemented with 1% (wt/wt) chrysin (CHY group) for 28 days followed by a single intraperitoneal injection of LPS (10 mg/kg BW). Control mice were given normal saline and all animals were kept for 72 hours before sacrifice. The results showed that chrysin attenuated histological injury of intestine and the production of pro‐inflammatory cytokines, TNFα, IL‐6 and IL‐1β. When compared to the LPS group revealed at genus level, CHY increased the proportion of Alloprevotella and Lachnospiraceae NK4A136, and decreased Escherichia_Shigella and Akkermansia. Furthermore, LPS group showed lower species richness of gut microbiota compared to the control animals, and the reduction was reversed by the addition of chrysin. In conclusion, chrysin can attenuate the LPS‐induced intestinal damage in mice. This protective effect appears to be mediated, in part, through beneficial changes to the microbiota.

Support or Funding Information

This study was supported by the National Science Council Grant 105‐2320‐B‐468‐001 Taipei, Taiwan.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Posttreatment Thymoquinone Can Reverse High‐Dose, Atorvastatin‐Induced Renal Injury in Rats
Authors: Hassan Sherif Sabry , Youakim Magdy Fouad , Aboulhoda Basma Emad
Published: June 2021

Background

Atorvastatin (ATO) is an HMG‐CoA reductase inhibitor used to lower blood cholesterol, but it causes renal injury in high doses, including the kidney. The active ingredient of Nigella sativa, Thymoquinone (TQ), has antioxidant, antiapoptotic, and antiinflammatory properties.

Aim of study

The aim was to investigate whether posttreatment TQ could reverse ATO‐induced renal injury, and the possible mechanism of action by which TQ produces such effect.

Methods

Forty adult male Sprague Dawley rats were divided into 4 groups: control, TQ‐treated, ATO‐treated, and combined ATO/TQ‐treated. Rats were euthanized by cervical dislocation. Blood samples and kidneys were tested for kidney functions, oxidative stress, and apoptosis markers and for histopathological examination. Data was expressed as mean ± standard deviation (SD) and categorical variables are presented as percentage. One‐way analysis of variance (ANOVA) and post hoc tests (Bonferroni test) were used for the comparison of mean values between groups. P values of less than 0.01 were assumed to be significant (P<0.01). Data analysis was done using Study data were analyzed using SPSS.

Results

The ATO‐treated group showed an increase in kidney functions, decreases in the oxidative stress markers, reduced glutathione and catalase, and increases in the protein carbonylation, lipid peroxidation, and caspase 3 activity. The ATO/TQ‐treated group appeared to improve these pathological changes. Morphometric analyses of the histopathological and ultrastructural examination supported these results.

Conclusion

Posttreatment TQ seemed to ameliorate oxidative stress‐induced renal injury produced by high‐dose ATO, suggesting a potential clinical application to the administration of high‐dose ATO, in patients with renal insufficiency.

Support or Funding Information

Self‐Funded

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Visualizing Cellular Adaptations during Ozone‐induced Lung Inflammation
Authors: Aulakh Gurpreet K , Singh Jaswant , Snead Elizabeth
Published: June 2021

Ozone (O3) is a prevalent environmental pollutant with well‐documented acute and chronic health effects. Neutrophils are known as the prime orchestrators of lung immune surveillance. Although bacterial infections are known to elicit a robust lung neutrophil and alveolar macrophage response, it is not known why and how neutrophils are activated in response to sterile lung injury. Acute exposures to 2 ppm O3 induce lung macrophage and neutrophil extravasation and airway hyper‐reactivity at 24 h, however, these concentrations are manifold higher than ambient O3 concentrations, thus precluding any understanding of the initial events. Our pilot experiments established ozone's dose and time dependent killing of alveolar macrophages and concomitant recruitment of leukocytes. Our data indicated that 50 ppb of O3 induces cell death in half of the lung alveolar macrophage population. Thereafter, we exposed C57BL6 mice to either filtered air or 50 ppb of O3 for 2 h to understand the kinetics of murine ozone induced acute lung injury. We report acute vascular neutrophil concentration and activation at as early as 2 h post exposure which continues up to 24 h after ozone exposure. Immunofluorescent staining revealed CD11b positive lung alveolar macrophage and neutrophil recruitment. In addition to conventional bronchoalveolar lavage, lung vascular perfusate and peripheral blood analysis, we present novel application of simple, yet powerful, configuration of a fixed‐stage wide‐field upright fluorescent intravital microscopic set‐up to investigate ozone's oxidative lung damage. We observed extensive alveolar actin filament disorganization (Fig. 1), an initial drop (at 2 h post‐O3 exposure) followed by restoration (18 h post‐O3 exposure) of alveolar reactive oxygen species (ROS) (Fig. 2) as well as mitochondrial potential (Fig. 3) in live animals. Important findings from these preliminary experiments include CD11b predominant phenotypes of pulmonary leukocytes, alveolar necrosis, actin disorganization, acute alterations in alveolar ROS production as well as mitochondrial potential. Thus, we present evidence of systemic as well as lung toxicity at 40 fold lower O3 concentrations. The findings are important in establishing a sensitive model of ozone induced acute lung injury.

Support or Funding Information

Fedoruk Centre and Innovation Saskatchewan.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Externally‐Applied Talc Containing Baby Powder Particles Can Enter the Female Reproductive Tract, Become Lodged In Pelvic Tissues, and Be Found In Ovarian Cancer Resections
Authors: Johnson Kurt E. , Popratiloff Anastas , Godleski John J
Published: June 2021

We used differential interference contrast (DIC), polarized light microscopy (PLM), and scanning electron microscopy with energy dispersive X‐ray analysis (SEM/EDS) to study both control talc‐containing baby powder (TCBP), a magnesium silicate, as well as specimens obtained from TCBP‐exposed human patients with ovarian cancer. Control TCBP specimens consist of a mixture of large and small birefringent particles (1–65 μm diameter), most of which are approximately isodiametric with aspect ratios averaging 1.3, but a substantial fraction of particles are elongated, with aspect ratios averaging 11 (Figures 1 and 2, note scale bar in Figure 1 applies as well to Figure 2). Detailed analysis of 200 randomly chosen particles from TCBP reveal three main classes: the most common class, (small aspect ratio and small area); and, two other uncommon classes; one with a small aspect ratio and large area; and, the other with a small area but large aspect ratio (Table 1).

In the SEM/EDS, both isodiametric and elongated particles show oxygen, magnesium, and silicon signals of talc (Figures 3 and 4). In biopsies from TCBP‐exposed patients with ovarian cancer, a subset of particles in malignant tissues studied with PLM revealed birefringent particles, commonly associated with macrophages (Figure 5). When paraffin‐embedded blocks of these tissues were examined with SEM/EDS, some particles gave nontalc signals, but some gave signals with peaks for oxygen, magnesium, and silicon in ratios indicative of talc (Figures 6 and 7). These results show that particles of TCBP, when applied externally, can make their way through the female reproductive tract and become embedded in malignant ovarian tissue. In addition, we found that most of the talc particles in malignant ovarian tissue were small, with diameters in the 4 μm range, interestingly, the most frequent class of particles in control TCBP.

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 World of the Vascular Surgeon: Knowledge of Anatomical Landmarks for Successful Surgical Outcomes
Authors: Sajan Abin , Torre Barrett , DiRaimo Robert , Caceres Noel , Marquez Samuel
Published: June 2021

To prevent strokes in at‐risk patients, the vascular surgeon may choose from a variety of procedures to counter the accumulation of atheromatous plaque in the carotid arteries: open surgery in the neck with carotid endarterectomy (CEA), carotid stenting with less invasive endovascular techniques, and the more recently developed trans‐carotid artery revascularization (TCAR). The height of the carotid bifurcation (HCB) is an important landmark for the surgeon because it aids in determining the specific surgical approach used, while seeking to minimize the risk to cranial nerves and other vital structures. The HCB is classically defined in relation to the cervical vertebral levels, hyoid bone, thyroid cartilage, and the angle of the mandible. Substantial variability exists across population groups generating extensive research to identify the precise vertebral levels linked to HCB with respect to ethnic and gender differences with a focus on left/right asymmetries. Although well reported, the relationship between cervical vertebral levels and the HCB is of little use during operative procedures because the patient's cervical spine is not readily accessible in the surgical field. Thus, we attempt to elucidate the HCB using anatomical landmarks available while operating. Anatomical dissections were performed on the right and left sides of the neck in five cadavers and morphometric measures of surgical landmarks were obtained. The following ratios were calculated: medial border of the clavicle (MBC) to HCB/MBC to mastoid process (MP), MBC to superior thyroid artery (STA) origin/MBC to MP, and MBC to facial vein (FV) origin/MBC to MP. In addition to the first reported measurements of the HCB from these accessible landmarks, the study found there are statistical significant differences between the right and left sides of the neck among and between individuals. The MBC to bifurcation, MBC to STA origin, and the MBC to FV origin ratio was greater on the left side with 0.74 vs 0.64 (p= .008), 0.70 vs 0.64 (p= .02), and 0.69 vs 0.61 (p=.004), respectively. Our preliminary study addresses the lack of surgically relevant anatomical measures surrounding the carotid region and takes a step toward establishing useful anatomical ratios that may contribute to successful surgical outcomes.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Weight losing on gastric restriction in an animal model
Authors: Akamatsu Flávia Emi , Itezerote Ana Maria , Saleh Samir , Hojaij Flávio , Andrade Mauro , Martinez Carlos Augusto Real , Jacomo Alfredo Luiz
Published: June 2021

Background

Obesity and its related comorbidities have detrimental effects for the affected individual and pose a major challenge on public health systems worldwide. The number of non‐surgical options to treat obesity are limited and the long‐term success of dietary or life style interventions is minimal Bariatric surgery leads to a long‐term reduction in body weight and in obesity‐related morbidity and is currently the only treatment modality with a proven mortality benefit. In this regard, research with animal models has significantly helped to elucidate some of the potential mechanisms underlying bariatric surgery. There is no information about gastric restriction with phytobezoar in animal model and the weight loss.

Purpose

Measure the weight loss until 10 days after surgical gastrostomy and compare to sham and normal group

Methods

Experimental model of gastric restriction was devised using rats. The animals were submitted to surgical gastrostomy and a cylindrical loofah is inserted into the stomach. We studied 30 adult male Wistar rats divided in three groups: the stomach reduction group (R10), the sham group (S10), which underwent the same procedure except for the loofah insertion and the control group (C10). Animals were fed and kept in separate cages. They were weighed every other day until being sacrificed on day 10. The results will always be described by the mean value with respective standard deviation. Comparison of multiple groups was performed using analysis of variance (ANOVA) followed by Bonferroni post hoc tests. All tests were applied with 5% significance level.

Results

The weight decreased significantly 9% in the first three days and then increases 6.2% until the weighing of third day and keep until the tenth.

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 Visfatin on Metabolic Profiles of an Animal Model of Type 2 Diabetes
Authors: Adeghate Ernest , AlShamsi Thuraya , Amin Amr , Hameed Rashed
Published: June 2021

Novel adipocytokines including visfatin present differentially in obese patients and are suggested to influence the development of type 2 diabetes in the background of obesity by modifying how insulin performs its function. Visfatin, in contrast to adiponectin was considered a harmful adipocytokine. This study examines the effect of visfatin administration on body weight, blood glucose, glucose tolerance and islet morphology in Goto‐Kakizaki (GK) rats, an animal model of type 2 diabetes mellitus. Visfatin (4 ng/Kg body weight) was given intraperitoneally to adult GK rats for 8 weeks. Control rats received an equal amount of the vehicle. Body weight and glucose level were measured every 2 weeks. Glucose tolerance test was performed at the end of the experiment. Immunofluorescence staining was performed to identify the pattern of distribution of visfatin in pancreatic islet cells of GK rats treated with or without visfatin. Co‐localization study was performed to determine whether visfatin resides with either, insulin, glucagon, somatostatin or pancreatic polypeptide (PP) in the islets of Langerhans. Immunofluorescence study shows that many islet cells in pancreatic islets of Langerhans of GK rats contain visfatin, where it co‐localizes mainly with insulin. The number of visfatin and insulin‐containing cells in pancreatic islets of GK rats treated with visfatin increased significantly (p < 0.05). Although most of the visfatin‐positive cells contain insulin, some islet cells located in the periphery of pancreatic islets also contain visfatin. Treatment with visfatin did not significant increase weight gain in GK rats compared to non‐treated control. Glucose handling was markedly (p < 0.05) improved in GK rats treated with visfatin. In conclusion, visfatin may play a role in the regulation of glucose metabolism.

Support or Funding Information

This work was supported by UAE University Research Grant # 02‐13‐11/08.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
VERANY: Cephalopods of the Mediterranean Sea: (1847)–1851, Genova (France)
Authors: LE FLOCH‐PRIGENT Patrice , NGUYEN Juliette , GILLOT Jean‐Bernard
Published: June 2021

The Alain Brieux's Bookstore owned in 2018 a rare, original monography written in Nice about the cephalopods of the mediterranean sea. It was published in Genova by the “Imprimerie des sourds‐muets”. The exact title was “Cephalopodes de la Méditerranée. Mollusques méditerranéens observés, décrits et chromolithographiés d'après le vivant. Première partie”. The book was an in‐quarto (in‐4). There were 132 pages and 43 plates. The drawings and prints were the work of the author. Jean‐Baptiste VERANY (1800–1865) discovered several species of cephalopods. The logigopsis (chirotheutis) veranyi holds his name. He collected a collection of natural history by his own since 1820. He performed a scientific cruise in South America on the Euridice, a royal sardinian vessel and after worked in Genova. He gave his collection in 1846 to the town of Nice (south coast of 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.
Topographic Anatomic Mapping of the Superior Cervical Ganglion with Novel Optical Clearing Method (LIMPID)
Authors: Thiyagarajah Nishanth H , Liu YeHe Hsee , Jenkins Michael Hsee , Lewis Stephen J , Hsieh Yee‐Hsee
Published: June 2021

BACKGROUND/AIM

The Superior Cervical Ganglion (SCG) contains cell bodies that project to various structures within the head and neck, including the nasopharynx, larynx, and submandibular glands. With the emergence of electrical stimulation of peripheral nerves as a treatment for disease pathologies, an emphasis must be placed on the anatomic mapping of end‐point structures. Topographic location within the ganglion as well as post ganglionic projections from the SCG are not well defined as of yet. The 2 main postganglionic sympathetic branches that exit the SCG are the Internal Carotid Nerve (ICN) and the External Carotid Nerve (ECN). The main focus of this work is to utilize a novel optical clearing method to create a topographic map of the SCG, as well as identify the anatomic pathways that these cell bodies travel to their end‐point structures.

METHODS

Male adult Sprague Dawley rodents (n=18) were anesthetized with Sevofluorane. These rats were separated into 3 equal groups; (1) Tract tracer injections with Hamilton micro syringe (1 μl DiI (8.3mg/ml)) in submandibular gland, (1 μl DiO (8.3mg/ml)) in nasopharynx, and (0.25–0.5 μl DiD (8.3mg/ml)) in larynx; (2) Unilateral surgical transection of the left ECN with bilateral SMG injections of 1μl DiI (8.3mg/ml) (3) Unilateral surgical transection of the left ICN with bilateral nasopharynx injections of 1μl DiO (8.3mg/ml). The SCG was harvested 7 days later through whole body perfusion/fixation with 1X Phosphate buffered saline (PBS)/4% Paraformaldehyde. SCG was placed in a clearing solution for 3‐D topographic mapping for visualization method called lipid‐preserving index matching for prolonged imaging depth (LIMPID). Imaging was performed using a Leica TCS SP8 gated STED.

RESULTS

Our results show distinct locations of cell populations for SMG, larynx, and nasopharynx respectively. In addition, the anatomical connection from SCG to SMG was ablated almost entirely or entirely following surgical ECN transection (>95% of ganglion cell population absent between transected and control lateralities in all experiments). Transection of the ICN nerve was also shown to entirely prevent the appearance of positive cells projecting to the nasopharynx unilaterally.

CONCLUSION

Our findings represent a novel topographic mapping of the SCG, and identify the postganglionic branches where cell bodies project to the nasopharynx and submandibular glands specifically. As bioelectronics moves forward, these anatomic mappings will be essential in the identification of intervention points for optimal therapy.

Support or Funding Information

Supported by NIH1OT2OD023860‐01

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Human Submandibular Salivary Gland Morphology Associated to Transcription and Growth Factors from Fetus to Aging
Authors: Gomes Tamires Albuquerque , Araújo Jose Roberto , Arias Vitor , Neves Adriana Costa , Silva Marcelo Cavenaghi Pereira
Published: June 2021

Saliva carries out several functions in the digestive system like the protection of the oral cavity and the beginning of carbohydrate digestion. This liquid is mainly produced by the submandibular gland as it is responsible for 60% of total saliva flow. This gland is characterized as acinar tubule composed, and during the fetal period, it develops in five stages, which are molded through high proliferative indices of the components. However, the aging process changes the morphology and function of this gland.

Transcription factors such as Nanog and Signal transducer and activator of transcription 3 (Stat‐3) regulate the process of proliferation, ckit and musashi‐1 induce cell proliferation while epidermal growth factor (EGF) is secreted by the submandibular gland and it has combined action to the Stat‐3. Considering the importance of these transcription factors and the role of EGF, the present study evaluated immunohistochemical expression of EGF, Nanog, Ckit, Musashi‐1 and Stat‐3 in submandibular glands of human fetuses between 12 and 18 gestational weeks, in adults (40 to 55 years old) and senile (over 85 years old). It was also evaluated quantitatively the morphological changes in components during development using Image J and statistical softwares. This study was approved by the Research Ethical Committee of UNIFESP, research number 618131.

The present study showed significant expression of all the transcription factors in the acinar system as in the ductal system during the stages of development. Also showed differences in the number of components between the adult and senile glands. Considering these results, we reinforce that the morphological changes observed in the submandibular gland is in consonance with the levels of the studied markers and the proliferation pattern during aging.

Support or Funding Information

This study was financed in part by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior ‐ Brasil (CAPES) Finance code 001 and also by FAPESP process number 2015/03967‐5.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modeling the Development of Cleft Lip and Palate in Variable Clefting Mouse Strains
Authors: Green Rebecca M. , Leach Courtney L. , Guo Sihan , Diewert Virginia , Marcucio Ralph S. , Hallgrimsson Benedikt
Published: June 2021

Epigenetic modifications in A‐strain mice can lead to cleft lip and palate (CLP). All A‐strain mice have a retrotransposon present near the Wnt9b gene, yet, two A‐strain mice cleft at very different rates. A/WySn clefts at 25% while A/J clefts at 5%, yet each line isogenic. The difference in clefting rates is thought to be due to changes in methylation of the retrotransposon. This variation in the penetrance of clefting among strains makes this an interesting model for human orofacial clefting as CLP is also variably penetrant in humans. Here, we test the hypothesis that these mice have subtle variations in 3D cell proliferation and apoptosis that cause variation in the shape of the facial tissues during crucial steps of facial formation, leading to CLP. Further, we predict these changes in proliferation, apoptosis and morphology correlate with Wnt9b levels. Genome changes between related A‐strain mice with different CLP levels also suggest that other loci may be involved in the variable clefting present in these mice. Combined, these results imply that variation in gene expression within a genotype leads to variation in cellular dynamics and the resulting morphogenesis, producing CLP leading to cleft development in a subset of A‐strain mice.

Support or Funding Information

NIH NIDCR R01 DE019638 to RS and BH. CIHR Foundation Grant to BH and RS. CIHR Postdoctoral Fellowship to RG.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interactions Between Channel Function, Cellular Differentiation, and Morphology in the Mandibles of Two Connexin‐43 Mutant Mouse Models
Authors: Moore Alyssa C , Jewlal Elizabeth , Barr Kevin , Laird Dale W , Willmore Katherine E
Published: June 2021

Background

Mutations in the gene encoding connexin‐43 (Cx43) can result in the human disease oculodentodigital dysplasia (ODDD), which has a strong craniofacial phenotype, including microphthalmia, a narrowed nose, and an increased incidence of cleft palate. The literature on the human condition of ODDD documents a mandibular phenotype including macrognathia and widened alveolar ridges; however, the mandibular phenotype has yet to be studied in detail in mouse models of ODDD. As well, a stepwise link between the amount of functional Cx43 protein, cellular differentiation, and phenotype has been found in heterozygous and homozygous Cx43 knockout mice, but this stepwise progression in Cx43 functionality, cellular changes, and phenotypic changes has yet to be investigated in mouse models of ODDD. Therefore, the purpose of this project is to determine the relationship between Cx43 function, celular differentiation, and mandibular morphology.

Methods

We used two mouse models with mutations in the gene encoding for Cx43 which have 20% channel functionality and a severe ODDD phenotype (Cx43G60S/+) or 50% channel functionality and a moderate ODDD phenotype (Cx43I130T/+), respectively. Homologous landmark data were collected from microCT images of newborn hemimandibles and analyzed with geometric morphometric analyses to quantify and visualize mandibular shape changes between mutants and wildtype littermates.

General mandibular histomorphology will be qualitatively assessed using Safranin‐O/Fast Green staining and compared among genotypes. We will also quantify expression of chondrocyte, osteoblast, osteoclast, and osteocyte differentiation markers in these mutant mice and their wildtype littermates using qRT‐PCR. With these procedures, we hope to gain an understanding of the relationship between Cx43 function, cellular differentiation, and mandibular morphology.

Preliminary Results

Cx43G60S/+ mice display significant alterations to mandibular shape and size when compared to their wildtype littermates. The changes to shape and size cluster around the incisor alveolar process, coronoid process, angular process, condylar process, and mental and mandibular foramina in Cx43G60S/+ mice. Potential drivers for these size and shape changes include delayed ossification and the interaction of different modes of ossification in secondary cartilage.

Significance & Future Directions

Our preliminary work confirms that altered Cx43 function results in altered mandibular morphology. Ongoing studies are focused on determining if cellular differentiation is altered in the bone and cartilage of our mutant mandibles and if these cellular changes can explain the phenotypic changes observed.

Support or Funding Information

This study was funded by NSERC.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Status of MMP 9 and TIMP 1 in Pregnant women with Preeclampsia in Indian Population
Authors: ARORA PALLAVI , MOCHAN SANKAT , GUPTA SUNIL KUMAR , SAXENA SHOBHIT , RANI NEERJA , PALLAVI SK , SHARMA ARUNDHATI , LUTHRA KALPANA , DWIVEDI SADANAND , BHATLA NEERJA , DHINGRA RENU
Published: June 2021

Introduction

Matrix metalloproteinases (MMPs) specifically MMP 9 is a key regulator of vascular and uterine spiral artery remodelling and its activity is controlled at multiple levels, including gene transcription, activation of its latent forms and endogenous inhibition by tissue inhibitors of metalloproteinases [TIMPs (specifically TIMP 1)]. Alteration in MMP 9 and TIMP 1 expression may contribute to uterine and vascular dysfunction leading to adverse pregnancy outcomes such as Preeclampsia (PE) and FGR. Therefore, in the present study, we sought to analyse the levels of this enzyme (MMP 9) and its regulator (TIMP 1) in the patients of PE.

Methods

The blood samples from the enrolled pregnant women [Total: 60, Preeclamptic: 30, Normotensive, Non‐proteinuric (Controls): 30)] were used for determining the mRNA expression of MMP 9 and TIMP 1 using qRT‐PCR. The sera of the above subjects were used for measuring MMP 9 and TIMP 1 protein levels using commercially available ELISA kits (R and D system). However, 30 caesarean delivered placentae (15 PE and 15 normotensive) were used to analyse the mRNA expression of MMP 9 and TIMP 1 using qRT‐PCR. Also, the MMP 9 and TIMP1 protein expression was seen by immunohistochemistry (IHC) and immunofluorescence (IF) staining. Zymography was performed for MMP 9 characterization and TIMP 1 expression was evaluated using western blot in these placentae.

Results

MMP 9 mRNA expression reduced in both plasma (2.66 folds) and placentae (6.66 folds) of PE patients as compared to controls. However, TIMP 1 mRNA expression was elevated in both plasma (2.23 folds) and placentae (8.32 folds) of PE patients as compared to controls. ELISA results demonstrated decrease in MMP 9 (p=0.0526; Wilcoxon matched‐pairs signed rank test) and increase in TIMP 1 (p=0.9133; Wilcoxon matched‐pairs signed rank test) protein levels in PE patients as compared to healthy controls. Immunolocalisation revealed stronger expression of MMP 9 in placental syncytiotrophoblasts and stroma in control placentae as compared to PE placentae. However, TIMP 1 intensity reduced in placental villi and decidual cells in the placentae from normotensive non‐proteinuric pregnant women. MMP 9 activity (zymography) was also found to be less and TIMP‐1 (western blot) upregulated in PE placentae as compared to control placentae.

Conclusion

Understanding the role of MMP 9 and TIMP 1 in uteroplacental and vascular remodelling and function could help design new approaches for prediction and management of preeclampsia.

Ethical Clearance: Ref. No. IECPG‐101/21.03.2018

Support or Funding Information

F.No.IR‐516/2018/RS

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Maternal Alcohol Exposure Augments Fetal Skeletal Anomalies in Diabetic Mouse Pregnancy
Authors: Rengasamy Padmanabhan , Shafiulla Mohammed
Published: June 2021

Introduction

There is a several‐fold increase in birth defects in diabetic pregnancy. Alcohol is an established‐teratogen in humans. Both alcohol and diabetes mellitus (DM) are independent risk factors for congenital anomalies. However, their teratogenic mechanisms are unknown. Diabetic women also might consume alcohol during pregnancy.

Aim

Our aim was to determine the effects of potential interaction between maternal alcohol and maternal DM during gestation on fetal development.

Methods

We made pregnant mice diabetic by a single dose (IP) of the diabetogenic compound streptozotocin (STZ) administered on gestation day (GD) 2. Controls received a proportionate volume of vehicle (citrate buffer). Groups of DM and non‐DM animals received ethanol (IP) in single doses of 0.03 ml/g body weight (25% v/v ethanol in normal saline) on GD 7 or 8. One group of DM animals had a daily dose of 6–8 IU of insulin subcutaneously. We collected the fetuses on GD 18, fixed in 95% ethanol and stained their skeletons with Alizarin red‐S and Alcian blue and observed for skeletal anomalies.

Results

Both ethanol and DM independently induced a low incidence of skeletal defects in fetuses. Ethanol injection of DM mice on GD 7 resulted in a moderate accentuation of DM‐related skeletal defects. Treatment of DM animals with ethanol on GD 8 resulted in severe skeletal defects, which affected regions of the axial and appendicular skeletons differentially. The combination treatment affected most markedly skeletal ossification in about 60% of animals. This treatment also caused an augmented frequency of skeletal variations (e.g. supernumerary ribs and sternal anomalies) and poor ossification of the bones of fore‐, and hind paws.

Discussion and Conclusion

Gestation day 8 was particularly susceptible to augmentation of skeletal defects with ethanol treatment. These data also suggest that alcohol‐diabetes interaction affects the ossification processes of mouse embryos resulting in extensive skeletal variations and malformations.

Support or Funding Information

Supported by (1) College of Medicine and Health Sciences, UAE University, Al Ain, UAE and (2) School of Medicine, University of Texas Rio Grande Valley, Edinburg, TX

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exploring Sources of Variation in Inter‐observer Reliability Scoring of Facial Expressions Using the ChimpFACS
Authors: Molina Alexandra , Florkiewicz Brittany , Cartmill Erica
Published: June 2021

The Facial Action Coding System (FACS) is currently the most popular systematic and standardized method of studying facial expressions in mammals. Since its original release for humans, the FACS has expanded to include primates such as chimpanzees. The FACS trains its users to analyze facial expressions by identifying individual muscle movements (called action units or AUs) that are combined to create facial expressions (or AU combinations). All researchers must pass a species‐specific certification test and are then considered reliable coders. After certification, interobserver reliability is rarely performed. However, reliability may be affected by variables like video length, apex identification, AU combination composition, and sociality effects. We analyzed the effects of these variables on reliability and propose that researchers should predetermine action units of interest for apex choice to reduce variation between observers. Two ChimpFACS certified researchers independently coded facial expressions from 30 hours of video of 17 chimpanzees (Pan troglodytes) housed at the Los Angeles Zoo. Coding reliability was then evaluated using Wexler's ratio (the method of calculating FACS inter‐observer reliability on the certification test). Preliminary results indicate that AU combination composition and sociality effects could have significant effects on coding reliability. We argue that researchers should predetermine which AUs to code as notable to facial expressions of interest and only code one individual during a time of high sociality to improve reliability in studies involving FACS coding. Employing these additional steps would improve replicability in facial expression studies, thereby strengthening research on the evolution of facial communication in primates.

Support or Funding Information

This research was supported by the National Institute of General Medical Sciences of the National Institute of Health under award number R25GM055052 awarded to T. Hasson. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. This project could also not have occurred without the support of the Los Angeles Zoo.

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