bluuebunny/crossref_metadata_2025 huggingface dataset:
Soluble Mediators Derived from Lactobacillus rhamnosus GG (LGG) Decrease Visceral Pain Hypersensitivity Induced by Early Life Stress
Authors: Neufeld Karen‐Anne McVey , O'Mahony Siobhain M , Waworuntu Rosaline V , Manurung Sarmauli , Gross Gabriele , Berg Brian M , Dinan Timothy G , Cryan John F
Published: November 2023

Maternal separation of rat pups is a robust and reliable model of early life stress that induces long‐term alterations to behavior and brain neurochemistry. These changes are particularly apparent with respect to the microbiota‐gut‐brain axis where probiotic feeding has been shown to ameliorate some of these stress‐induced alterations. It is possible that biologically relevant products of probiotic metabolism are involved in such effects, and therefore could deliver the required benefits without requiring the live organism to be present. In this study the effects of a specific preparation of soluble mediators from the probiotic Lactobacillus rhamnosus GG (LGG) were evaluated. Previously, immunomodulatory properties of this material have been demonstrated both in vitro and in vivo. Production conditions have been standardized with cultivation and downstream processing steps such as desalting, sterile filtration and lyophilization. The maternal separation protocol was conducted as described in O'Mahony et al., 2009. Rats were separated from their mothers for 3 hrs/day from postnatal day (PND) 2 to 12. Starting at weaning (PND 21), both non‐separated (NS) and maternally separated (MS) offspring were randomized into separate experimental groups and were provided drinking water with or without supplementation of LGG soluble mediators. Supplementation of the lyophilized materials was benchmarked to the number of LGG viable cells during cultivation to give a dose equivalent to a range from 1.3×108 to 4.3×108 viable LGG/animal/day throughout the study until PND 85. Visceral hypersensitivity was assessed with the colorectal distension test conducted at PND 79. There were no differences in body weight or water and food intake across groups. MS rats demonstrated visceral hypersensitivity to colorectal distension reflected by a lower pain threshold when compared to NS rats, which was ameliorated by the LGG soluble mediators (p=0.03). There were no differences in the number of observed pain behaviors between treatment groups. Interestingly, the increased pain threshold in rats that received LGG soluble mediators was not observed in animals that received unconditioned culture medium subjected to the same processing steps as the LGG soluble mediators. This indicated the presence of specific bioactives in the culture medium secreted by LGG during growth. In conclusion, administration of LGG soluble mediators in rats reversed the altered visceral pain sensitivity induced by MS. Further studies are needed to elucidate specific mechanism of actions related to gut and brain neurochemistry.

Support or Funding Information

Supported by Science Foundation Ireland and Mead Johnson Nutrition.
Inulin Effect on Weight Loss and Associated Parameters with The Development of Cardiovascular Disease in Obese Adults
Authors: Castro‐Sanchez Francisco Humberto , Ochoa‐Acosta Dora Alicia , Valenzuela‐Rubio Nancy Guadalupe , Dominguez‐Rodriguez Maribel , Fierros‐Valdez Jesus Antonio , Vergara‐Jimenez Marcela
Published: November 2023

Obesity is one of the most concerning public health problems in Mexico and it has being associated with the development of cardiovascular diseases (CVD). A healthy diet has demonstrated to improve weight loss and blood lipids but also the inclusion of functional foods could be used. Inulin, a non‐digestible carbohydrate, has been shown to have the ability to promote weight loss and improve the lipid profile associated with CVD.

The aim of the present study was to examine the effect of addition of inulin plus a moderate carbohydrate diet on weight loss and parameters associated with cardiovascular disease.

Sixteen obese dislipidemic subjects were recluted and distributed in a double‐blind, crossover trial for 18 weeks with two eight‐week treatment periods (9g of inulin/day or 9g of dextrose/day as placebo), separated by a two‐week washout period.

Anthropometric and lipid profile were recollected at the beginning and the end of each treatment. Both treatments significantly improve weight loss and others anthropometric parameters (BMI, Body fat and also waist and hip circumferences) without statistical difference between them. Inulin significantly increased HDL cholesterol (p<0.05) and tend to reduce triglycerides.

Non‐significant changes were observed on Total and LDL cholesterol. In conclusion, we can suggest that the addition to inulin and a moderate carbohydrate diet tend to reduce cardiovascular risk by increasing HDL cholesterol and reduction of triglycerides concentrations.
Effect of Konjac Noodles Intervention in Patients with Type 2 Diabetes
Authors: Li Sing‐Chung
Published: November 2023

Non‐communicable diseases (NCDs), which include diabetes, cardiovascular diseases, chronic obstructive pulmonary diseases and cancers had become an emerging health problem in the world. According to the 2014 World Health Organization Global Status Report, NCDs caused 38 million (68%) of all deaths worldwide. Among NCDs, diabetes was also an important cause of 1.3 million deaths in 2010, twice as many as in 1990. Prevalence of diabetes in Taiwan had increased two fold over ten years. Approximately 90% of patients belong to type 2 diabetes around the world. Our data suggested konjac glucomannan noodles (KGN) as a staple food in a 4 weeks period intervention could be decreased in body weight, body mass index (BMI), and waist with statistical differences except glycosylated hemoglobin (HbA1c). The object of this study was to investigate whether prolong KGN intervention could improve more clinical biomarkers in type 2 diabetes patients. This study was a positive‐controlled, randomized, single‐blind, and parallel‐group clinical trial. All 42 patients were recruited from Saint Mary's Hospital Luodong, 4 patients were refused and 2 patients were excluded by clinical indicators. Matched 36 subjects were randomized to non‐KGN or KGN for a 12‐week period. Participants completed twice 24‐hour dietary recalls at baseline and again at 12‐week follow‐up. Compared between the non‐KGN group intervention, the changes of body weight (0.21%; p=0.766), BMI (0.11%; p=0.878), waist (0.41%; p=0.430) and HbA1c (−4.53%; p=0.128) were no statistical differences. However in KGN group, body weight (−1.04%, p=0.022), BMI (−1.11%, p=0.026), waist (−2.89%, p=0.003) and HbA1c (−6.63%, p=0.036) were significantly decreased. In conclusion, consumption konjac glucomannan noodles as a staple food may have clinical benefits associated with the achievement of body weight, BMI, waist and HbA1c in type 2 diabetes patients.
Oral administration of hydrolyzed red ginseng extract improves learning and memory capability in cognitive‐decline models
Authors: JU SUNGHEE , Jang JinHo , Kim MinSun , Jeong GaIn , Seo HyeLin , Woo YaeJin , Lee Seung‐Kwon , Oh Jisun , Kim Jong‐Sang
Published: November 2023

Korean red ginseng (Panax ginseng C.A. Meyer) contains various pharmacologically active constituents including ginsenosides. Recently we found that red ginseng extract has a strong antioxidant activity and that pectinase‐mediated hydrolysis augments its radical‐scavenging activity. In addition, the content of compound‐K, a bioavailable and bioactive ginsenoside, was significantly increased in the hydrolyzed red ginseng extract compared to the non‐hydrolyzed extract. We further investigated neuroprotective and cognitive‐enhancing effects of the hydrolyzed red ginseng extract in vitro and in vivo. To induce oxidative stress‐induced neurotoxicity, mouse hippocampal HT22 cells and C57BL/6 mice were exposed to an excess glutamate and D‐galactose, respectively. For behavioral assessments, the passive avoidance, Y‐maze, and Morris water maze tasks were performed following treatment with the extracts or compound‐K. Our results from the in vitro and in vivo tests demonstrate that (1) red ginseng extract (containing compound‐K) protects neuronal cells from oxidative damage through the induction of Nrf2 and antioxidant enzymes, (2) the learning and memory impairments induced by oxidative stress were alleviated by treatment with the hydrolyzed red ginseng extract, (3) the hippocampi (particularly cell counts and nuclear arrangement) of the stressed mice was histologically pathologic, which was rarely observable from the hippocampi of the extract‐treated mice. Our findings suggest that red ginseng extract hydrolyzed by pectinase may effectively alleviate oxidative stress‐mediated memory deficits possibly through Nrf2‐related detoxification.

Support or Funding Information

This work was supported by the Food Functionality Evaluation program funded by the Ministry of Agriculture, Food and Rural Affairs through Korea Food Research Institute.
Methylene Chloride Fraction of Salicornia herbacea L. Suppresses Glutamate‐induced Neuronal Cytotoxicity and Improves Galactose‐induced Learning and Memory Impairment
Authors: Kim MinSun , Kang SuJin , Jo GwangRae , Jang JinHo , Jeong GaIn , Ju SungHee , Seo HyeLin , Woo YaeJin , Oh Jisun , Kim Jong‐Sang
Published: November 2023

Salicornia herbacea L., also called as glasswort, is a halophyte growing in salt marshes and has been used as a seasoned vegetable and a traditional medicine for various disorders. It is a nutrient‐rich herb containing plenty of bioactive compounds such as selenium, flavonoids. In this study, we in vitro examined if ethanol extract from glasswort is neuroprotective under oxidative stress‐induced environment. Hippocampal HT22 cells were treated with 5 mM glutamate in the presence of ethanol extract from glasswort or its subfractions in five different solvents (hexane, methylene chloride, ethyl acetate, butanol, and water). Ethanol (EtOH) extract and methylene chloride‐soluble subfraction (MC fraction) of the herb were effective in protecting HT22 cells from glutamate‐induced toxicity. In addition, EtOH extract and MC fraction induced intracellular antioxidant enzyme NQO1's activity and inhibited ROS production. The levels of Nrf2 and its downstream antioxidant enzymes were significantly elevated by both EtOH extract and MC fraction. We further investigated whether MC fraction is protective against galactose‐induced cognitive decline in animal model. C57BL/6J mice were fed MC fraction, and intraperitoneally injected with D‐galactose on a daily‐basis to generate oxidative stress and cause learning and memory impairments. After 6 weeks, behavioral and biochemical tests were conducted. The group fed MC fraction decreased escape latency time, reduced DNA damage and lipid peroxidation, and enhanced NQO1 enzyme activity in tissues, compared to D‐galactose‐treated control group. Overall, these findings suggest that MC fraction of glasswort extract could be a promising agent for AD prevention.

Support or Funding Information

This work was supported by the Food Functionality Evaluation program funded by the Ministry of Agriculture, Food and Rural Affairs through Korea Food Research Institute, and by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (Project No. 2013R1A1A2013362).
Ascorbic Acid Content, Phenolic Compounds and Antioxidant Capacity of Brazilian Savannah Native Fruits
Authors: Naves Maria Margareth Veloso , Alves Aline Medeiros , Dias Tiago , Hassimotto Neusa Mariko Aymoto
Published: November 2023

The Brazilian Savannah (Cerrado biome) produces several native fruits, such as cagaita (Eugenia dysenterica DC), cerrado cashew (Anacardium othonianum Rizz.) and gabiroba (Campomanesia adamantium (Cambess.) O. Berg), which are widely used as food and in folk medicine. In the present study, these fruits were investigated as potential sources of antioxidant substances. The fruits were characterized concerning the total ascorbic acid content by liquid chromatograph equipped with diode‐array detector (HPLC‐DAD), total phenolic compounds by UV/visible spectrophotometry, antioxidant capacity by three different methods (2,2‐diphenyl‐1‐picrylhydrazyl [DPPH], Ferric Reducing Antioxidant Power [FRAP], and Oxygen Radical Absorbance Capacity [ORAC]), and the flavonoids composition by HPLC‐DAD. The gabiroba presented higher contents of ascorbic acid (614.9 mg.kg−1) and total phenolics (12,225.9 mg GAE.kg−1), and greater antioxidant capacity than cagaita and cerrado cashew. Nevertheless, the three fruits showed considerable contents of bioactive compounds in relation to other Brazilian native fruits. The correlation coefficients between total ascorbic acid or total phenolic contents and antioxidant capacity were high and significant (0.970 ≤ r ≤ 0.998; P < 0.05). Quercetin derivatives were found in cagaita (19.4 mg.kg−1) and myricetin derivatives in cerrado cashew fruit (12.0 mg.kg−1). The gabiroba showed relevant catechin content (233.2 mg.kg−1). Besides, peonidin 3‐glucoside was the main anthocyanin in cerrado cashew fruit, which flavonoids composition was first identified in this study. These Brazilian native fruits may be considered as important sources of natural antioxidants and potential alternatives of bioactive compounds for food science and nutrition. The gabiroba (C. adamantium) stands out from other fruits because of its relevant content of vitamin C and of phenolic compounds, particularly catechin, and because of its high antioxidant capacity.

Support or Funding Information

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq‐Brazil) for financial support (Proc. # 485565/2012‐9) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES‐Brazil) for scholarship support.
Elasticity and Vascular Endothelial Function in Patients with Aortic Valve Insufficiency
Authors: Silveira João Vicente , Katayama Keyla Yukari , Sangaleti Carine Teles , Ferreira Janaina , Hong Valeria Costa , Lopes Heno Ferreira , Consolim‐Colombo Fernanda Marciano
Published: November 2023

Backgroud

Aortic insufficiency (regurgitation) is a common valve disease in young and middle‐aged individuals, which requires from the cardiovascular system a continuous volume overload. Patients can remain asymptomatic for many years and subsequently they can present symptoms of congestive heart failure. The remodeling of the great vessels has been observed in patients with severe regurgitation. However, little is known about the changes in the elasticity of small arteries and endothelial function in a moderate condition, without clinical symptoms.

Aim

Our goal was to assess small artery elasticity and endothelial function by peripheral arterial tonometry. The hemodynamic profile was characterized by the continuously and noninvasively analyses of the blood pressure curves.

Methods

We evaluated 41 individuals of both gender, aged 18–55 years divided into two groups: control (n = 23 healthy) and aortic regurgitation (n = 18, classified as mild to moderate through Doppler echocardiography criteria, and asymptomatic). For the acquisition and analysis of hemodynamic variables: systolic blood pressure (SBP), diastolic (DBP), heart rate (HR), end‐systolic volume (ESV), cardiac output (CO) and total peripheral resistance (TPR), the monitor Finometer® and the BeatScope software was used. The vascular elasticity and endothelial function were assessed by the EndoPAT®. Statistical analysis was performed using Student's t test, unpaired, with p <0.05, the values are presented in mean ± SEM.

Results

Individuals with aortic regurgitation (48.5 ± 3.2 years; BMI 27.8 ± 1.4 kg/m2) compared to the control group (n = 23, 41.7 ± 1.8 years; BMI 26 4 ± 0.7 kg/m2) had the follows hemodynamic changes: increased SBP (138.6 ± 3.7 vs 126.7 ± 2.8 mmHg) ESV (151.7 ± 13.5 vs 83.9 ± 3, 0 ml) and CO (10.6 ± 1.0 vs 5.4 ± 0.2 l/min); and decreased DBP (64.7 ± 2.6 vs 75.3 ± 1.8 mmHg) and TPR (0.6 ± 0.1 vs 1.1 ± 0.1 UPR). Endothelial function was not different between groups. However, there was a significant increase in the elasticity of small arteries in the aortic regurgitation group in absolute values (20.2 ± 5.0 vs 5.7 ± 3.9%) and also for normalized values according to a HR of 75 bpm (16.9 ± 4,5 vs −2.5±3.9%).

Conclusion

Young adults with asymptomatic aortic regurgitation had a higher elasticity of the small arteries and a preserved endothelial function.
Animal Models of Heart Failure: the Forgotten Animal Model
Authors: Chandrasekera Charukeshi
Published: November 2023

Heart failure remains a leading cause of death and an economic burden worldwide. The research community continues to tackle this epidemic by dedicating extensive research efforts to delineate heart failure mechanisms and to develop novel therapeutics. Animal models form the cornerstone of these basic and translational research efforts; however, despite decades of extensive research, human heart failure remains poorly understood and only limited therapeutic measures are available to human patients. While animal models in species ranging from fruit flies to primates have considerably advanced our understanding of animal cardiovascular physiology and pathophysiology, research success from these models have not proportionally translated into therapeutic success for human patients. A growing body of human‐specific mechanistic and clinical data indicate that the translational discrepancy is primarily due to the species barrier: from gene expression to excitation‐contraction coupling to drug responses, animal models do not accurately replicate human heart failure causes, natural history, manifestations, complications, and responses to treatments. Immutable interspecies differences occur at every level of biological activity and are further confounded by biological variability (age, sex, and strain), and even differences in housing and husbandry practices as well as experimental conditions. Given the disappointing history of animal research translation for heart failure, the paucity of definitive treatments, and the emergence of heart failure as a dominant cause of morbidity and mortality in the U.S. and across the world, the need to move from animal models to human‐specific research seems evident. In fact, other researchers in the field are urging the heart failure community to “modify and enhance the currently dominant translational paradigm and provide new important directions of research.” Here we present evidence indicating why it is imperative for the heart failure research community to devise a plan to incorporate Homo sapiens as the quintessential animal model. We provide specific examples of how genetic, biochemical, physiological, anatomical, and technical limitations inherent in animal models can be overcome with similar studies in humans and present how an integrative human‐based research framework incorporating every level of human biological complexity – from nucleic acid to whole organism level and beyond – can serve as the primary platform from which basic science discoveries are launched and more effective therapeutics are tested. If the ultimate goal of the research community is to advance preventive and reparative approaches for human heart failure, we need to transition from the quest to generate “better” animal models to the prioritized, widespread use of human‐based, human‐relevant research strategies.

Support or Funding Information

Author Institution
Cardiac compensation in ErbB2‐deficient zebrafish embryos
Authors: Fleming Nicole D , Samsa Leigh Ann , Liu Jiandong
Published: November 2023

During heart development, the heart chambers undergo dramatic changes in chamber topology, most notably the formation of luminal, muscular protrusions called trabeculae. While altered trabeculation is associated with many congenital heart diseases, complete failure to form trabeculae results in embryonic lethality across different species. We have previously reported that ErbB2 signaling is required for cardiac trabeculation. Although erbb2 mutant hearts do not exhibit any functional defects initially, these hearts develop progressive cardiac dysfunction with reduced fractional shortening, and eventually fail. However, the mechanism underlying this functional deterioration in the absence of trabecular formation is not known. Using zebrafish as a model, we found that inhibition of ErbB2 activity pharmacologically and genetically to block cardiac trabeculation resulted in an increase in ventricular cardiomyocyte cross‐sectional area and myofibril size. This cardiac phenotype is reminiscent of hypertrophic growth of an adult heart subjected to mechanical overload. Consistently, we found that the expression of nppa, a hypertrophic marker gene, was elevated in erbb2 mutants compared to controls. In addition, inhibition of Target of Rapamycin (TOR) signaling by rapamycin treatment suppressed erbb2 mutant hypertrophic growth phenotype, suggesting that hypertrophy of erbb2 mutant hearts occurs in an mTOR‐dependent manner. Taken together, we propose that trabeculae may play a role in optimizing chamber topology for efficient cardiac function.

Support or Funding Information

National Science Foundation Graduate Research Fellowship (NSF‐GRF): DGE‐1144081
Phospholamban is localized in autophagosome in dilated cardiomyopathy mouse model, TgPLNR9C mice
Authors: Yokoe Shunichi , MacLennan David H. , Asahi Michio
Published: November 2023

An Arg9 to Cys point mutation in phospholamban (PLN) is the leading cause of dilated cardiomyopathy (DCM) in both humans and in transgenic mice (TgPLNR9C mice). Our earlier study of PLN showed that impaired autophagic flux in HECT domain and Ankyrin repeat‐containing E3 ubiquitin protein ligase 1 (HACE1)‐null mouse hearts was associated with increased PLN protein levels. Here, we show that HACE1 expression was decreased in 8‐, 16‐weeks old TgPLNR9C mouse hearts, and PLN was co‐immunoprecipitated with the autophagic adapter protein, p62, and confocal immunofluorescent microscopy revealed intracellular co‐localization of PLN with p62. At the same time, electron microscopy showed increased autophagosome in TgPLNR9C mouse hearts. These results suggest that PLN is degraded by autophagy, and an impairment of the autophagy pathway promotes PLN accumulation in TgPLNR9C mouse hearts. Furthermore, we also observed decreased expression of the von Hippel‐Lindau (VHL), another E3 ubiquitin ligase in TgPLNR9C mouse hearts. Taken together, we propose that diminished expression of E3 ubiquitin ligases, such as HACE1 and VHL could be related to the progression to DCM in TgPLNR9C mice.
The effect of Body Mass Index on Pain Levels of Patients with Fibromyalgia
Authors: Torres‐Gutiérrez César J. , Merriwether Ericka , Rakel Barbara , Dailey Dana , Muenters Li Alemo , Abdelhamid Ramy , Darghosian Leon , Vance Carol , Crofford Leslie , Sluka Kathleen
Published: November 2023

Fibromyalgia (FM) is a chronic pain syndrome characterized by widespread pain, depression, and mood and sleep disturbances. There is currently no consensus on the pathogenesis of FM. Overweight women are 60–70% more likely to develop fibromyalgia and experience more severe pain symptoms than normal women. Adipose cells release pro‐inflammatory cytokines (IL‐6, TNF‐α, Leptin) that are linked to pain pathogenesis. This study aims to determine the relationship between weight or body mass index (BMI) and pain in fibromyalgia. A sample of 93 women was recruited for participation in a set of tests which classify the pain felt by the participants. Pearson Product Moment Correlation (r) analyses were done between BMI and the pain levels. One‐way analysis of variance (ANOVA) was used to determine group differences in pain measures. For a sub‐analysis consisting of 27 patients, blood samples were acquired from the participants. The Peripheral Blood Mononuclear Cell layer (PBMC) was isolated. Monocyte phenotype was determined using fluorescence‐activated cell sorting (FACS) and a Pearson Product Moment Correlation (r) analysis was done. We found that there is no relationship between BMI, Pain levels and Monocyte phenotype.

Support or Funding Information

This study was supported in part by the National Institutes of Health (NIAMS) AR063381 and AR063381S1, and the Carver College of Medicine Institutional funds. We thank the FAST study team for their assistance with scheduling and data collection.
Multi Quantum Biophotonic Energy's Ability to Reduce Fear Response and Anxiety in Williams Syndrome (case study)
Authors: Cohen Jay , Einstein George , Tulp Orion L
Published: November 2023

Due to the deletion of multiple genes on the long arm of chromosome 7, individuals with Williams Syndrome often elicit a hyperactive response to nonsocial frightening stimuli. The objective of this research was to determine if Multi Quantum Intravenous UVC/UVA light energy and subsequent NIR were effective in reducing the anxieties and fear responses associated with these stimuli. Our study involved a 17 year‐old female with Williams Syndrome who expressed distress when exposed to the sounds of laughter. We determined the particular sounds and associated frequencies that evoked anxiety and a fear response with this individual. These sounds were recorded and when reproduced elicited a consistent and predictable hyperactive response from this individual. Cognitive Behavioral Therapy, CBT, alone did not significantly reduce the hyperactive response recorded.

This individual received Multi Quantum Intravenous UVC/UVA energy followed by Quantum NIR. The amount of energy induced was calculated for a 36.28Kg individual.

This treatment was administered once daily over a 4 day period. The individual was then exposed to the same recordings that previously elicited a hyperactive response. Subsequent responses were noted to be diminished and less consistent for several days following the Multi Quantum Biophotonics treatment. (MBPT). Observational data from this study suggested that MBPT can help modulate the limbic’ system's fear response that is initiated in individuals with Williams Syndrome.

Further investigational research involving EEG studies combined with the Avtandil Anuashvili method identifying specific brain functions with Williams Syndrome individuals is indicated.
How the γPKC activity is regulated in neurodegenerative spinocerebellar ataxia type 14 (SCA14)
Authors: Aslam Naveed , Alvi Farah
Published: November 2023

The neurodegenerative spinocerebellar ataxia type 14 (SCA14) is linked with slowly progressive cerebellar ataxia accompanied by abnormal eye movements and slow slurred speech. The ataxic symptoms are due to loss of purkinje cells leading to cerebellar neurodegeneration. Recent observations directly link γPKC with SCA14 and its gene PRKCG is now widely recognized as a disease causing gene. γPKC is specifically expressed in brain and is highly expressed in purkinje cells. The SCA14 disease is characterized by degeneration of purkinje cells. Experimental observations indicate that γPKC can modulate the morphology and behavior of purkinje cells. Both positive and negative regulation of γPKC can be linked with changes in size and complexity of dendritic arbor in purkinje cells. Here, we investigate the question that how the activity of γPKC is regulated in purkinje cells of cerebellum? The growth of dendritic structure is strongly inhibited by γPKC activation. Here, we examine a proposal that γPKC activity in purkinje cells can be regulated through a DAG‐mediated molecular loop between γPKC and DGKγ. Our results show that γPKC activity in purkinje cells is regulated through a net negative feedback loop. Our results imply that on purinergic receptor stimulation the γPKC is activated through a sequence of DAG‐mediated molecular events. However, the γPKC restricts its own activation through a net negative feedback loop. Through our approach we are able to clarify the mechanism behind the spatiotemporal interaction of DAG effector molecules i.e. γPKC and DGKγ.

Support or Funding Information

BioSystOmics
The Antioxidant Effects of Thymoquinone in Lipopolysaccharide (LPS)/Interferon Gamma (IFNg) Activated BV2 Murine Microglia Cells
Authors: Cobourne‐Duval Makini K , Taka Equar , Soliman Karam F.
Published: November 2023

Chronic activation of microglia, the resident macrophages of the central nervous system (CNS), is implicated in the pathogenesis and progression of neurodegenerative disorders such as Alzheimer's disease. Disproportionate microglial activation drives a vicious self‐perpetuating neurodegenerative cycle via the excessive release of reactive oxygen species and pro‐inflammatory cytokines in the system. We explored the antioxidant effects of thymoquinone (TQ), a major bioactive component of Nigella sativa seed oil, in microglial activation. We hypothesized that the antioxidant properties of TQ are mediated in part by reducing the expression of proteins involved in microglia‐derived oxidative stress and modulating the activity of antioxidant enzymes. In this study, BV2 cells were stimulated with E. coli lipopolysaccharide (LPS) & interferon gamma (IFNg) in the presence or absence of TQ. To profile the effect of TQ on the global protein expression of BV2 microglial cells, two‐dimensional (2D)‐gel electrophoresis with MALDI‐TOF/TOF (matrix‐assisted laser desorption/ionization – time of flight) analysis was employed. Additionally, we assessed TQ's effect on microglial production of superoxide and nitric oxide, superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) enzyme activities and as well as the ratios of oxidized:reduced glutathione (GSSG:GSH) indicative of overall oxidative stress levels. The results obtained indicate that TQ caused changes in the expression of 60 kDa heat shock protein in LPS/IFNg stimulated BV2 cells as well as significantly (p<0.001) decreased the production of superoxide and nitric oxide and significantly (p<0.0001) decreased the levels of oxidative stress. TQ also modulated the activities of the antioxidant enzymes demonstrating increased antioxidant capacity. These findings suggest that TQ may be useful in delaying the onset or progression of neurodegenerative disorders involving chronic activation of microglia and the resultant excessive oxidative stress in the CNS. Thus, intervention of microglial activation process could be a promising therapeutic target for the treatment of a number of neurodegenerative conditions.

Support or Funding Information

NIH Grants : NIMHD G12 MD007582 and P20 MD006738
FACT: Functional Analysis Clustering Toolbox for visualizing disease progression
Authors: Yang Ruoting , Watson Daniel , Hammamieh Rasha , Jett Marti
Published: November 2023

In recent years, many studies collected temporal gene expression profiles based on DNA microarray technology for predicting the disease progression or host response to an external stress. Monitoring the functional changes, time dependency and noise is crucial to be able to dissect gene patterns. The popular computational packages for time‐series clustering usually combine several traditional clustering methods; however, these methods are not designed for time‐series and neglect the temporal information. This approach is precise and easy to apply, but it is sensitive to noise. As a result, we developed a novel computational tool ‐ FACT (Functional Analysis Clustering Toolbox) ‐ to identify the major gene patterns, visualize and compare pathways, and track functional progression. FACT can dissect spike patterns over time, provide the underlying biological functions for these spikes, and link them into a network.

To illustrate the effectiveness of FACT, we applied it to eight Prion disease datasets of global gene expression in the brains of eight distinct mouse strain ‐ prion strain combinations throughout the progression of the disease. We identified the major response timeline for the prion infection for each mouse‐prion strain. Several important pathways, such as the complement pathway, showing strain‐ and time‐ specific changes, have been teased out and compared to the original paper. We identified the major responsive pathways; the complement pathway was upregulated after week 12, and the response of the complement pathway was negatively correlated with active prion levels, consistent with the incubation time.

In conclusion, FACT provides a new rich, robust resource for diagnostic biomarker and therapeutic target discovery. This application to prion datasets illustrates its powerful capacity for dissecting time‐ and strain‐specific responsive genes and pathways and uncovering both the common functional progression and the unique functions for certain strains.

Support or Funding Information

DISCLAIMERS: Research was conducted in compliance with the Animal Welfare Act and all other Federal Requirements. The views expressed are those of the authors and do not constitute endorsement by the U.S. Army.
BAC‐based Molecular Cytogenetics: from Blood to BAC in Just a Week
Authors: Weier H.‐Ulli G , Weier Jingly F , Zeng Hui , Lu Chung‐Me , Wang Mei
Published: November 2023

For over two decades, our laboratories have been involved in translational research by designing nucleic acid probes and assays for karyotype analysis of mammalian cells and tissues. Due to its clinical significance, most probes and assays were developed for the analysis of human specimens ranging from oocyte and sperm to zygotes and in vitro fertilization embryos, fetal and extra‐embryonic tissues to somatic cells from childhood and adulthood tumors. The goals of our probe development efforts aimed at assays that provide the highest possible specificity, outstanding signal‐to‐noise ratios and excellent reproducibility, while minimizing the cost of multiplexed assays. Initial developments targeted gender determination of preimplantation embryos and/or the detection of numerical or structural chromosome abnormalities using single cell, multi‐probe/‐color fluorescence in situ hybridi‐zation (FISH). Many of the early probes based on chromosome‐ or locus‐specific DNA fragments, oligonucleotides, PCR products and plasmid clones are now commercially available. However, when multiplexed assays interrogate multiple loci, the cost per assay or test is often beyond the capabilities of individual labs. In our recent developments, we took advantage of resources generated in the Intl. Human Genome Project, such as large DNA insert bacterial artificial chromosome (BAC) libraries and validated physical maps to develop breakpoint‐specific probes or chromosome enumerator DNA probes. Using BAC's as starting material, we are now able to generate specific DNA probes of unsurpassed hybridization quality for very affordable, complex but easy‐to‐apply assays in just a matter of days.
THE PROTEIN KINASE Cβ INHIBITOR, ENZASTAURIN, DECREASES AMPHETAMINE‐MEDIATED BEHAVIORS IN RATS
Authors: Altshuler Rachel , Gnegy Margaret , Jutkiewicz Emily
Published: November 2023

Amphetamines elicit their motivating effects by increasing extracellular dopamine levels in the brain. Extracellular dopamine levels are regulated by the dopamine transporter (DAT), a transmembrane protein that transports dopamine from the extracellular space into the cell. Amphetamines are taken up by DAT and reverse the transporter to release dopamine into the synapse. One protein that has been shown to be important for the mechanism of amphetamine's action is protein kinase C β (PKCβ). The inhibition of PKCβ reduces amphetamine‐stimulated dopamine efflux through DAT in vitro and in vivo while having no effect on dopamine uptake. While the effects of PKC inhibitors on amphetamine‐stimulated dopamine efflux have been characterized, it is important to understand the effects of PKCβ inhibition on amphetamine‐stimulated behaviors. The goal of this study was to determine whether inhibition of PKCβ will decrease amphetamine‐stimulated locomotion and amphetamine self‐administration. For all experiments, the selective PKCβ inhibitor enzastaurin was administered to male Sprague‐Dawley rats by intracerebroventricular (i.c.v.) injection 18 hours prior to evaluating amphetamine‐mediated behaviors. Locomotor activity was measured in infrared beam break boxes following administration of a single dose (1 mg/kg) or cumulative doses (0.32–5.6 mg/kg) of amphetamine. In self‐administration studies under a fixed ratio 5 (FR5) schedule of reinforcement, rats earned infusions of amphetamine (0.032 mg/kg/infusion) or sucrose pellets in 60‐ or 20‐min daily sessions, respectively. Pretreatment with enzastaurin reduced amphetamine‐stimulated locomotion at lower amphetamine doses (0.32 and 1 mg/kg), but this effect was surmounted by larger doses (≥ 3.2 mg/kg) of amphetamine, demonstrating a rightward shift in the amphetamine dose effect curve for locomotion. In amphetamine self‐administration studies, enzastaurin decreased the number of amphetamine infusions by over 60%. This effect was specific for amphetamine because enzastaurin did not alter the number of sucrose pellets earned. This study demonstrated that PKCβ inhibition attenuated amphetamine mediated behaviors, likely in a surmountable fashion. PKCβ inhibitors may be useful therapeutic targets for decreasing amphetamine use and abuse without altering natural rewards. Future work will investigate the mechanisms by which PKCβ inhibition alters amphetamine‐stimulated behaviors.

Support or Funding Information

Funded by DA11697 and T32‐GM007767.
Antagonism of the Rate‐Decreasing Effects of Abused Psychostimulants and Bath Salt Component Mephedrone in Rats
Authors: Saber Iman A. , Walker Ellen A.
Published: November 2023

To compare the potencies to decrease ongoing behavior among the monoamine alkaloid derivatives of the amphetamine class and the synthetic cathinone mephedrone, 12 food‐restricted male Sprague‐Dawley rats were trained to respond on a fixed ratio 5 schedule of reinforcement for banana‐flavored sucrose pellets. Dose‐response curves for the behavioral effects of d‐amphetamine, methamphetamine, 3,4‐methylenedioxy‐methamphetamine (MDMA), and mephedrone were determined in a multiple‐trial, cumulative dosing procedure. In addition, the rate‐decreasing effects of these psychomotor stimulants were examined in combination with dopamine (DA) antagonists to assess the role of subtypes of DA receptors in the rate‐decreasing effects of the psychostimulants.

When multiple doses of DA1 antagonist SCH23390 were combined with a moderate dose of methamphetamine (1.0 mg/kg), an antagonism was observed. However, SCH23390 also appeared to have dose‐dependent, rate‐decreasing properties of its own potentially explaining the observed combined rate‐decreasing effects with the lower and higher doses of the stimulants.

The DA2/3 antagonist, sulpiride, slightly attenuated the effects of low and moderate doses of both d‐amphetamine and methamphetamine. When combined with MDMA and mephedrone, attenuation occurred at low and moderate doses, respectfully.

These results support previous work demonstrating the similarities and differences between these commonly abused CNS drugs in respects to dopamine activation. Furthermore, this dopaminergic comparison may apply to novel substituted amphetamines and provide insight for mechanisms underlying the pharmacological effects of novel cathinone derivatives such as mephedrone.
Effects of Intermittent or Ad Libitum Consumption of High‐Fat Diet on the Reinforcing Effects of Cocaine in Male Rats
Authors: Collins Gregory T , France Charles P
Published: November 2023

Drug abuse and obesity are serious public health problems. Dopamine systems play a central role in mediating the reinforcing effects of drugs and food, and the prolonged use of drugs or consumption of foods high in fat is known to alter the function and/or sensitivity of neurotransmitter systems, including dopamine. The current studies evaluated the impact of ad libitum or intermittent (1‐hr per day; Mon, Wed, Fri) consumption of the high‐fat diet on the acquisition of cocaine self‐administration during late adolescence (PND 52–62), relative to rats that consumed a standard diet (n=10–12 male rats per condition). During adulthood (PND ≥70), dose‐response curves for cocaine self‐administration were obtained under both fixed ratio (FR) 5 and progressive ratio (PR) schedules of reinforcement. Although the rate of acquisition was comparable across the three dietary conditions, high levels of responding (≥65 infusions of 0.032 mg/kg cocaine per session) were observed more frequently (85%) in rats with intermittent access to the high‐fat food compared with rats eating only the standard (60%) or high‐fat diets (20%). Similarly, when dose‐response curves for cocaine self‐administration were evaluated under an FR 5 schedule of reinforcement, intermittent consumption of the high‐fat diet shifted the dose‐response curve leftward and/or upward relative to the dose‐response curve obtained in rats that consumed the standard diet; ad libitum consumption of the high‐fat diet did not alter the cocaine dose‐response curve. Although dose‐response curves obtained when cocaine was self‐administered under a PR schedule did not differ among the three groups of rats, a subset of rats from the intermittent high‐fat condition completed significantly more ratios than all other rats. In addition to self‐administering more cocaine under the FR 5 schedule and completing larger ratios to obtain cocaine under the PR schedule, this subset of rats also consistently consumed more high‐fat food during the 1‐hr access period than other rats from the intermittent high‐fat condition. Although it is unclear what factors account for these differential levels/patterns for consumption, the current results provide additional evidence that the abuse‐related effects of drugs, such as cocaine, can be influenced by dietary factors and suggest that patterns of food consumption (i.e., intermittent versus ad libitum) might have a greater impact than the type of foods that are consumed on vulnerability to drug abuse.

Support or Funding Information

CPF is supported by a NIDA Senior Scientist Award (K05 DA017918).
Characterization of Mephedrone and its Bioactive Metabolites
Authors: Mayer Felix P , Partilla John S , Wimmer Laurin , Seddik Amir , Burchardt Nadine , Schmid Diethart , Bulling Simon , Ecker Gerhard F , Mihovilovic Marko D , Baumann Michael H , Sitte Harald H
Published: November 2023

The psychostimulant 4‐methyl‐N‐methylcathinone (mephedrone, MEPH) is a major representative of former legal highs, which were sold as “bath salts”. Despite the ban of MEPH in 2011, the drug is still widely abused for recreational purposes and as doping agent. MEPH manipulates monoaminergic neurotransmission in an amphetamine‐like fashion. Therefore, MEPH acts as a substrate of the high‐affinity monoamine transporters (MATs) for dopamine (DAT), serotonin (SERT) and norepinephrine (NET) and triggers transporter‐mediated release of neurotransmitters. Interestingly, drug abuser reports state that the action of MEPH resembles the action of 3,4‐methylenedioxymethamphetamine (“ecstasy”, MDMA). However, MEPH acts as non‐selective substrate at MATs whereas MDMA clearly favors SERT over DAT and NET. We sought to test the phase‐1 metabolites of MEPH for their activities at MATs; which may be responsible for the observed impact on serotonergic neurotransmission.

Radiotracer‐flux experiments were applied to determine the interactions between MATs and the tested metabolites in vitro and ex vivo. Single‐cell FRET measurements were performed to monitor test‐drug induced conformational changes in MATs in real‐time. Moreover, we assessed locomotion and stereotypy in vivo in rats and determined the extracellular concentrations of dopamine and serotonin by microdialysis in nucleus accumbens.

The phase 1 metabolites were found to inhibit uptake mediated by MATs with appreciable affinities in the low micromolar range and induced efflux in a dose‐dependent manner. Real‐time FRET measurements revealed an amphetamine‐like mode of action at MATs. Consistent with in vitro findings, the test drugs mimicked the parental drug in vivo with pronounced effects on extracellular serotonin.

The fact that MDMA‐like actions have been attributed to MEPH appeared to be inconsistent with its pharmacological profile. Our data unraveled the mode of action of MEPH and its phase‐1 metabolites on a molecular level and indicate that the metabolites essentially contribute to the action of MEPH.

Support or Funding Information

Austrian Research Fund/FWF grants F3506 and W1232 to HHS Intramural Research Program of the NIDA, NIH, grant DA000523‐07 to MHB FWF‐DK MolTag, FWF W1232 to MDMAustrian Research Fund/FWF grants F3502 and W1232 to GFE FPM is a recipient of a DOC‐fellowship of the Austrian Academy of Sciences
Behavioral and Pharmacological Effects of Taurine in Mice
Authors: Brown Taylor , McAvoy Kenneth , Susino John , Hindieh Jennifer , Hoefer William , Rossi Grace C
Published: November 2023

Taurine is a prominent and naturally occurring organic acid which has found its way into the diet of millions of people. Taurine is found in massive quantities within the energy drinks that have become increasingly popular in the last few decades. Though previous research has deemed taurine safe to consume, little is known about the effects of taurine in high doses or chronic doses. This study investigates the effect that various doses of taurine have on CD‐1 mice at both acute and chronic conditions. The motor coordination and activity levels of mice were observed for 20, 50, 100, and 200mg doses, which represented the acute condition, and also were observed after three weeks of ad libitum drinking of 1% or 5% taurine solution, which represented the chronic condition. The results showed that taurine has dose‐dependent effects that differ between levels of exposure. These findings indicate that acute doses of taurine may be beneficial in increasing bursts of movements, but repeated exposure to taurine may have adverse effects on coordination, overall movement time, and stereotypical movements.