bluuebunny/crossref_metadata_2025 huggingface dataset:
Astaxanthin Mediates Mitochondrial Dynamics in Mice
Authors: Shen Xinchun , Lyu Yi , Wu Lei , Lin Dingbo
Published: November 2023

Objective

Alteration of mitochondrial dynamics closely links to cellular oxidative stress and inflammation, key risk factors for the development of chronic diseases including obesity and diabetes. Bioactive astaxanthin has been found effective to antioxidation. The purpose of this study is to explore the functional roles of astaxanthin in regulation of mitochondrial dynamics and the consequence in energy metabolism in mice.

Methods

Six‐week‐old male β, β‐carotene‐9′, 10′‐oxygenase 2 (BCO2) knockout (KO) and the isogenic wild‐type C57BL/6J(WT) mice were fed an astaxanthin supplementation diet (0.04% astaxanthin supplemented to the AIN93‐maintenance diet) or a control diet (AIN93‐maintenance) for 8 weeks, respectively. On the 8th week of the study, mice were subjected to a phenotyping test. At termination of the study, blood and liver tissues were collected. Plasma metabolic biomarkers were tested, including cholesterol, lipoproteins, triglycerides, nonesterified fatty acids (NEFA), and uric acid. Fresh hepatic mitochondria were isolated and subjected to mitochondrial respiratory function assays. Immunoblotting was performed to determine proteins expression.

Results

Significant astaxanthin accumulation occurred in BCO2 KO mice. Astaxanthin supplementation significantly decrease the energy expenditure in WT mice, which was abolished by ablation of BCO2, suggesting that proper astaxanthin metabolism is vital to the phenotype. In addition to the astaxanthin‐elevated plasma NEFA levels in both WT and BCO2 KO mice, disruption of astaxanthin metabolism resulted in increases in the plasma levels of C‐reactive protein, total cholesterol, HDL, LDL, triglycerides, and uric acids in BCO2 KO mice. Mitochondrial respiration activity analysis results revealed the increases in basal respiration rate and proton leak in liver of BCO2 KO mice fed astaxanthin, compared to the AIN93 maintenance diet, though no obvious change was found in hepatic mitochondrial respiration in WT mice after astaxanthin treatment. Furthermore, elevated expression of phosphorylation of unc‐51 like autophagy activating kinase 1(ULK1) on Ser757, and uncoupling protein 2, mitofusin 2, and decreased protein levels of p62 and ATG13 indicating potentiated mitophagy and fusion in liver of WT fed the astaxanthin diet.

Conclusions

Astaxanthin‐potentiated mitophagy and fusion would link to decreased energy expenditure in wild type mice. Disruption of astaxanthin metabolism could be associated with metabolic disorders, including inflammation and dyslipidemia in BCO2 knockout mice.
Cobalamin‐deficiency results in altered partitioning of mitochondrial one‐carbon units to formate
Authors: MacMillan Luke , Tingley Garrett , Brosnan Margaret E , Brosnan John T
Published: November 2023

Vitamin B12, a co‐factor for methionine synthase, plays an important role in folate‐dependent remethylation of homocysteine. We have shown that cobalamin deficiency results in a marked elevation in plasma formate.

De novo production of formate in rats was determined as described by Morrow et al (JBC 2015). Formate production from one‐carbon precursors by isolated rat liver mitochondria was measured by GC‐MS. 14CO2 production from 3‐[14C]‐serine by isolated rat liver mitochondria, expression of key genes (RT‐qPCR) and changes in protein expression (immunoblot) were also determined.

Vitamin B12‐deficient rats (B12‐def) showed a 55% increase in endogenous formate production, consistent with the elevated formate levels. Formate production by isolated mitochondria from serine, dimethylglycine, and sarcosine was increased by at least 60% in B12‐def animals while 14CO2 production from serine by these mitochondria was 26% lower compared to replete controls. The expression of Mthfd1l in B12‐def animals was unchanged but the expression of Aldh1l1 and Aldh1l2 was reduced by 40–60% of that in the livers of control animals.

We suggest that the partitioning of mitochondrial formyl‐THF between oxidation to CO2 (by ALDH1L2) and production of free formate (by MTHFD1L) is a key control point in one‐carbon metabolism. In B12‐def the decreased flux through ALDH1L2, in the face of a constant rate of 10‐formyl‐THF disposition can explain the increased production and increased concentration of formate. We speculate that the decreased hepatic SAM levels found in B12‐def animals may cause increased production of one‐carbon groups in the form of formate to replenish hepatic SAM. Since mitochondrial formate becomes available to the cytosol we suggest that this metabolic bifurcation may regulate the entry of formate into the cytosolic one‐carbon pool.

Support or Funding Information

Supported by CIHR, RDC, and Memorial University of Newfoundland
Food Insecurity, Glycemic Control, and Heart Rate Variability among Latinos with Type 2 Diabetes
Authors: Bermúdez‐Millán Angela , Pérez‐Escamilla Rafael , Lampert Rachel , Segura‐Pérez Sofia , Damio Grace , Chhabra Jyoti , Wagner Julie
Published: November 2023

Food insecurity disproportionately affects Latinos. Whether food insecurity as a stressor, is associated with decreased heart rate variability (HRV), a non‐invasive measure of autonomic status which predicts poor cardiovascular outcomes, is unknown. This study examined the relationship between food insecurity and HRV among U. S. Latinos with type 2 diabetes. Secondary data analysis of baseline data from the CALMS‐D stress management trial was conducted. Participants completed surveys of food insecurity, demographics, and socioeconomic status. Participants wore a 7‐lead, 3‐channel ambulatory ECG monitor (Holters) for 24 hours. HRV indices included standard deviation of the R‐R interval (SDNN), and log‐normalized very low frequency (lnVLF) 0.0033 to < 0.04 Hz; low frequency (lnLF) 0.04 to <0.15 Hz; and high frequency (lnHF) 0.15 to 0.40 Hz. ANOVA and ANCOVA were used for statistical analyses. Of the 94 participants, 46 reported diabetes‐related food insecurity and 48 reported diabetes‐related food security. Participants who were diabetes‐related food insecure (“In the past year, has it happened to you that you did not have enough money to buy the food you should eat for your diabetes?”) had lower SDNN, lnVLF, lnLF, and lnHF, all p<.05, even after adjusting for glycemic control and socioeconomic hardship. Food insecurity is a powerful and unique form of social adversity. Efforts to address food insecurity may benefit diabetes outcomes.

Support or Funding Information

This study was funded by the National Institute of Minority Health and Health Disparities (grant number 5 R01 MD005879 to J.W. and R.P.‐E., principal investigators) and by the American Diabetes Association (7‐13‐TS‐31 to J.W., principal investigator).
The prevalence status of overweight and obesity in Chinese teenagers aged 15–17
Authors: Fang Hongyun
Published: November 2023

Center for Disease Control and Prevention, Beijing, China, People's Republic of Objective To analyze the prevalence of overweight and obesity in Chinese teenagers aged 15–17. Methods A total of 8 333 subjects aged from 15 to 17 years old were retrieved from China National Nutrition and Health Surveillance 2010–2012 and enrolled in this study(4 173 people in urban, 4 160 people in rural areas). Overweight and obesity were defined according to the WHO recommended BMI for age and sex in 2007. Age‐standardized results were calculated incorporating a complex sample weighting using the population data from the sixth national census conducted by the National Bureau of Statistics in 2010. Results The prevalence of overweight and obesity in Chinese teenagers aged 15–17 were 8.6% and 2.8% respectively, the sum of the two is 11.4%. The prevalence of overweight and obesity in male were 9.6%, 4.0% respectively, while 7.5%, 1.5% in female. Conclusions 11.4% of Chinese teenagers aged 15–17 are overweight or obese. China currently has more than 1/10 of 15–17 year olds overweight or obese, nearly 6 million people. The prevalence of overweight and obesity of boys is higher than that of girls, with city boys as the main risk group.

Support or Funding Information

The major projects of health care reform‐Monitoring of nutrition and health status of Chinese Residents (2010–2012), supported by National Health and Family Planning Commission (formerly Ministry of Health)
Morphologic analysis of spinal cord ischemia ‐ reperfusion injury caused by aortic cross clamping in a rat model
Authors: Bobinac Dragica , Celic Tanja , Vukelic Iva
Published: November 2023

Background

Spinal cord ischemia and paralysis of lower extremities are devastating postoperative complications that accompany surgery of aortic aneurysm. Spinal cord ischemia ‐ reperfusion injury (I/R) results in overproduction of reactive species leading to tissue oxidative stress, which impact the neuronal network in spinal cord as well as supporting white matter cells. Here, we report the longitudinal analysis of activity and number of neurologic cells in the area of aortic clamping.

Methods

Transient aortic occlusion was produced in rats by cross – clamping of the abdominal aorta for 45 minutes. Animals were sacrificed at several time points after reperfusion; 1, 6 and 48h; to determine time correlated changes in activity and number of neurologic cells in spinal cord. Spinal cord tissue was analyzed by immunofluorescence using specific primary antibodies, glial fibrillary acid protein (GFAP) and NeuN. Also, we wondered whether survival of neurons was related to their expression of nuclear factor erythroid 2‐related factor 2 (Nrf2), an oxidative stress protective protein.

Results

In the present study, ischemia – reperfusion injury of spinal cord was manifested by moderate loss of neurons in dorsal horn after 6h of reperfusion, and significant loss of neurons in dorsal horn after 48h of reperfusion compared to sham animals. Neurons located ventrally and laterally indicated no change in number. Surviving neuron cells, howerver, showed expression of Nrf2 with its nuclear translocation escpecially pronounced in neurons located in ventral horn. Longitudinal analysis of neuronal number showed that loss of neurons in dorsal horn was limited to lumbar part of spinal cord. Analysis of GFAP positive cells revealed moderate increase in number 6h post reperfusion in lateral part of spinal cord, while 48h post reperfusion this increase was even more pronounced compared to sham animals. Increase of number of GFAP positive cells was followed with astrocyte activation in lateral part of spinal cord presented as increased GFAP positive area in the same part of spinal cord. Longitudinal analysis of spinal cord revealed that astrocyte activation is present even in thoracic parts of spinal cord, higher than segments which were targeted by aortal clamping.

Conclusion

Our data indicate that I/R spinal cord injury induces changes in number of GFAP and NeuN positive cells and these changes correlate to duration of reperfusion time. The surviving neurons express Nrf2 in their nucleus, which shows the involvement of Nrf2 pathway in neuron survival.

Support or Funding Information

Grant approved and financed by Faculty of Medicine University of Rijeka
Developmental Localization of the Kv1.3 Voltage‐gated Potassium Channel and Insulin Receptor in Hypothalamic Nuclei Governing Energy Homeostasis
Authors: Doczi Megan Anne , Yacawych Warren
Published: November 2023

The Kv1.3 voltage‐gated potassium channel is expressed in various cell types associated with metabolic function including skeletal muscle, adipocytes, and neurons. Pharmacological Kv1.3 blockade has been shown to profoundly affect the regulation of body weight and energy homeostasis, suggesting that this channel may play a key role in regulating metabolic processes. In addition, the insulin hormone can suppress Kv1.3 ionic current through the tyrosine kinase activity of the insulin receptor (IR), suggesting a link between insulin signaling and Kv1.3 channel activity. Recent evidence from our laboratory has identified Kv1.3 gene expression in the embryonic avian hypothalamus, a brain region critical for maintaining metabolic homeostasis, although the specific protein localization remains unclear. The purpose of this study was to identify the localization of Kv1.3 and IR in the developing embryonic avian hypothalamus to establish a future model for in ovo insulin delivery, ultimately determining whether insulin affects the expression and function of Kv1.3 during development. Western blot analysis was used to validate monoclonal antibody binding for both the Kv1.3 channel and IR in embryonic day 12 (E12) avian whole brain lysates (n = 3). In accordance with the observed insulin‐dependent current suppression of the channel, we tested the hypothesis that Kv1.3 is localized to hypothalamic nuclei governing food intake and energy expenditure. Immunohistochemical analysis of hypothalamic brain sections from E12 embryos indicate that IR protein is localized to the ventromedial hypothalamus and arcuate nucleus (n = 3), while Kv1.3 channels are expressed more widely throughout this brain region, including enhanced staining within the subventricular zone (n = 3). This study demonstrates the hypothalamic expression of both Kv1.3 and IR in the developing avian embryo and suggests a role for voltage‐gated ion channel regulation in the physiological patterning of embryonic hypothalamic circuits. Further understanding the localization and function of insulin‐sensitive Kv1.3 channels during hypothalamic embryogenesis may provide insight into the developmental mechanisms of metabolic disorders such as diabetes and obesity.

Support or Funding Information

Support made possible by the Vermont Genetics Network through Grant Number P20GM103449 from the INBRE Program of the National Institute of General Medical Sciences (NIGMS) and the National Center for Research Resources (NCRR), components of the National Institutes of Health (NIH).
Television Food and Beverage Marketing to Children in Costa Rica: Is a Regulatory Policy Needed?
Authors: Corrales Irina Zamora , Jensen Melissa L. , Kroker‐Lobos Fernanda , Vandevijvere Stefanie , Ramírez Manuel
Published: November 2023

Background

Childhood overweight and obesity is a global public health concern, affecting 21.4% and 20.8% of Costa Rican children (5–12 years of age) and adolescents (13–19 years of age), respectively. Marketing of unhealthy foods and beverages (i.e. those high in saturated fat, sugar and/or sodium) has been shown to influence children's food preferences and consumption.

Little is known, however, regarding the scope of the problem in Costa Rica. This study aimed to examine the extent and nature of television food advertisements which children are likely to be exposed to, as well as the nutrition profile of the foods and beverages advertised in Costa Rica.

Methods

Television data were recorded for four weekdays and four weekend days, between 6:00 am and midnight, for four channels (two national and two cable), that were selected based on audience measurement data and a clear focus on children (i.e. cartoon channels). We obtained a total sample of 288 hours of television broadcasting, during the months of September and November 2016.

Results

Across all channels, 8983 advertisements (ads) were identified, of which 20.75% were for foods and beverages (F&B). Cable channels showed a higher number of F&B advertisements during weekends compared to weekdays. On average, the two national channels presented 70.9 (SD=19.2) F&B ads/day on weekdays, and 29.8/day (SD=16.2) on weekends; whereas the two cable channels resulted in a mean 57.8 (SD=16.6) F&B ads/day on weekdays and 74.6 (SD= 31.8) F&B ads/day on weekends. A total of 224 F&B items were advertised, many of which generally considered high in fat, sugar and/or sodium: fast‐food (10.71%), candy and chocolates (9.38%), sugar sweetened beverages (5.36%), snack foods (4.91%%), canned and dehydrated meal helpers and soups (4.91%), processed meats (4.02%) and sauces and dressings (4.02%).

Conclusions

Children and adolescents living in Costa Rica are exposed to television marketing of unhealthy foods and beverages daily. Due to the link between food marketing, preferences and consumption, our findings help justify the need for regulatory actions by national authorities.

Support or Funding Information

This study was supported by a research training grant from IDRC‐CRDI and CIIPEC‐INCAP
Assessing The Impact of The Diet on Cardiometabolic Outcomes: Are Two Consecutive Measures Post‐Intervention Really Necessary?
Authors: Allaire Janie , Talbot Denis , Couture Patrick , Tchernof André , Jones Peter JH , Etherton Penny K , West Sheila , Connelly Philip W , Jenkins David JA , Lamarche Benoît
Published: November 2023

Background

Important day‐to‐day variations in the blood concentrations of risk factors may lessen our ability to detect significant effects of any given nutritional intervention on cardiometabolic risk. In theory, relying on several consecutive measurements of a biomarker outcome should decrease variability and hence increase statistical power. However, there is very little empirical evidence supporting such a theory in nutrition research. The purpose of this study was to examine how using the mean of two consecutive measures vs. only one measure post treatment influences the capacity to detect a change in often‐used cardiometabolic risk factors in response to nutritional interventions.

Methods

We used data from two randomized double‐blind crossover trials that have assessed the impact of docosahexaenoic acid (DHA) on cardiometabolic risk factors, the first one in the context of a supplementation study and the second one in the context of fully‐controlled feeding conditions. We used equations and simulations to compare the effect of using the mean of two consecutive measures vs. only one measure post treatment on the capacity to detect a change in often‐used cardiometabolic risk factors in response to nutritional interventions.

Results

We found that the reduction in sample size or the gain in statistical power attributed to using the mean of two consecutive measures vs. a single measure is marginal for serum total cholesterol (C), non‐HDL‐C, HDL‐C, LDL‐C, apolipoprotein B100, triglycerides (TG) and C‐reactive protein, even when attempting to assess small effect sizes. For example, the difference in the required sample size to detect a medium effect size (e.g. 0.5) for the change in TG in response to supplemented DHA between using one vs. the mean of two consecutive TG measurements post treatment is 4 subjects (71 vs. 67 subjects respectively). For LDL‐C, this difference is 2 subjects (67 vs. 65 subjects).

Conclusions

These data indicate that the reduction in sample size and hence the gain in statistical power attributed to using the mean of two measures taken on consecutive days vs. a single measure is small and unjustified to detect a change in cardiometabolic risk factors in response to DHA in the context of crossover nutritional studies.

Support or Funding Information

JA is a recipient of PhD Scholarships from the Canadian Institutes of Health Research (CIHR) and Fonds de recherche du Québec – Santé (FRQ‐S).
Does the copulatory plug cause pseudopregnancy in female mice?
Authors: Lough‐Stevens Michael J , Dean Matthew D.
Published: November 2023

Female choice is the pre‐ and post‐copulatory responses of females to male phenotypes and behaviors. The neuroendocrine and anatomical responses of females to mating within the context of sexual selection is poorly characterized. One possible target of female choice is the copulatory plug, which is an epoxy formed of multiple seminal proteins post‐ejaculation in the vagina of many mammals. The protein TGM4 is an essential protein for forming the copulatory plug. TGM4 knockout male mice, although indistinguishable anatomically and behaviorally from wild type males, cannot form a copulatory plug, which makes them an ideal study system for understanding its function. Successful copulation and deposit of the copulatory plug is hypothesized to be required for successful implantation of zygotes in mice, a physiological state called pseudopregnancy. Females hours after receiving an ejaculation and copulatory plug from a male exhibit increased neuronal activity of the transcription factor c‐Fos in the ventromedial hypothalamus, a proposed site of sexual arousal in female rodents. Subsequently within days the uterine lining begins to thicken in anticipation of implantation of zygotes.

We hypothesize that pseudopregnancy may represent a previously unrecognized form of female choice. To characterize better the role of the copulatory plug in pseudopregnancy, we mated females against TGM4 wild type and knockout males. Pseudopregnancy in the brain was assessed through immunohistochemical assay of c‐Fos activity in the ventromedial hypothalamus. Pseudopregnancy in the uterus was assessed via non‐surgical injection of ConcavaninA beads, which mimic zygote size, into the uterus 2.5 days after copulation and examination of the uterus on day 4.5 for sites of implantation. Our study can perhaps offer insight into the neuroanatomical and uterine basis for female choice in female mice.

Support or Funding Information

Funded by the National Science Foundation (NSF).
Betulinic acid decreased de novo lipogenesis and lipid accumulation in differentiated 3T3‐L1 adipocytes
Authors: Kim Hyun Kyung , Go Gwang‐woong
Published: November 2023

Betulinic acid, derived from birch wood bark, is a naturally occurring pentacyclic triterpenic acid. Betulinic acid has diverse beneficial effects on cancers, inflammation, and obesity. Anti‐lipogenic effect of betulinic acid has recently been demonstrated; betulin interfered the maturation of SREBP1/2 in liver by inducing interaction of SCAP and insig. The action of betulinic acid in adipocyte, however, has not been investigated. It was hypothesized that betulinic acid would reduce de novo lipogenesis and/or adipogenesis in differentiated adipocytes. 3T3‐L1 was differentiated for 8 days, followed by betulinic acid treatment (1, 2, 5, and 10 μg/mL). Betulinic acid did not affect cell viability up to 10 μg/mL in differentiated adipocytes. Lipid accumulation assayed by oil red O staining was significantly decreased by 5 and 10 μg/mL of betulinic acid (p<0.01 and p<0.001). Intracellular triglyceride was likewise reduced by betulinic acid in a dose‐dependent manner (p<0.05). The level of critical lipogenic enzymes and its transcription factor were analyzed by using immunoblotting. Phosphorylation of acetyl CoA carboxylase was increased by betulinic acid in a dose‐dependent manner, resulting in diminished its activity. Fatty acid synthase was significantly decreased by 10 μg/mL of betulinic acid (p<0.05). Stearoyl CoA desaturase was clearly diminished by all concentration of betulinic acid (p<0.001). The matured SREBP1 (~68 kDa) was notably decreased by 10 μg/mL of betulinic acid (p<0.001). In conclusion, betulinic acid reduced de novo lipogenesis and lipid accumulation in differentiated 3T3‐L1 adipocytes. Therefore, betulinic acid, a natural triterpenic acid from birch bark, is a potent nutraceutical for anti‐obesity.
Attenuation of Obesity by Ornithine Enriched Blueberry Liquid Starter in 3T3‐L1 Preadipocyte and High‐Fat Diet Fed Obese Mouse Model
Authors: Oh Suk‐Heung , Kim Mina , Yu Ok‐Kyeong , Kim Su‐Gon , Cha Youn‐Soo
Published: November 2023

The primary objective of this study was to assess ornithine enriched blueberry liquid starter (OB) potential in inhibiting the adipogenesis process in 3T3‐L1 preadipocytes and attenuating obesity related parameters in high‐fat diet induced obese mice. OB did not show any cytotoxic effect, however, treatment with OB resulted in reduction in triglyceride accumulation compared to the blueberry liquid (BB) and non‐treatment control (C). Four‐week‐old C57BL/6J mice were fed with control diets containing 10% kcal from fat (CD) or high‐fat diets containing 60% kcal from fat (HD). Treatment groups were given high fat diet; the samples OB, BB, ornithine (OR) were administrated orally. Compare to the HD control group, the animals given OB showed significant reduction in body weight gain. Furthermore, obesity related parameters like serum lipid profiles were attenuated in the OB treated group. Overall, these results demonstrated the potential of OB in making processed food products which could be capable of attenuating obesity.

Support or Funding Information

This research was financially supported by the Ministry of Trade, Industry, and Energy (MOTIE), Korea, under the “Regional Specialized Industry Development Program” supervised by the Korea Institute for Advancement of Technology (KIAT).
Tributyltin exposure alters post‐embryonic growth and intestinal microbiota assembly in zebrafish
Authors: de la Torre Canny Sol Gomez , Mueller Olaf , Rawls John Franklin
Published: November 2023

Obesity is a ubiquitous public health problem that warrants the study of environmental factors that, by regulating energy balance, promote the storage of excess energy in adipose tissue. Two important environmental factors that promote adipose tissue size are microbial communities residing in the intestine, and environmental chemicals that may act as obesogens. We developed an in vivo model to test the obesogenic effect of chronic tributyltin (TBT) exposure in post‐embryonic zebrafish and to study the effect that this chemical has on intestinal microbiota. Zebrafish develop multiple adipose tissue depots, which consist of white adipocytes that display morphological and molecular homologies with those of mammals. Using fluorescent lipophilic dyes to measure adipose tissue in vivo, we found that microbiota colonization, as in mammals, promotes adipose tissue accumulation in zebrafish. Also, we show that 21‐day exposure to TBT at a nominal dose of 1 ug/l was sufficient to increase the growth of adipose tissue in zebrafish compared to control animals. Unexpectedly, TBT exposure also inhibited somatic growth as early as 14 days post‐exposure and at a dose as low as 0.1 ug/l. High‐throughput sequencing of 16S rDNA revealed specific effects of TBT on intestinal bacterial communities by 14 days post‐exposure, such as an increased Chitinibacter spp. abundance. Together, our results indicate that TBT exposure promotes adipose tissue deposition even in the absence of a high fat diet, and affects other distinct aspects of post‐embryonic growth and intestinal microbiota assembly. This work provides the first comprehensive analysis of the effect of a chemical obesogen on zebrafish somatic growth, adipose tissue accumulation, and intestinal microbiota composition, providing a critical frame of reference for future studies to test for mechanistic interactions between environmental toxins, microbiota, and animal hosts.

Support or Funding Information

NIH grants R21‐ES023369 and R24‐OD016761
Alleviation of paraquat‐induced oxidative stress and pulmonary fibrosis by betaine in rats
Authors: Na Jong Deok , Choi Yeo Jin , Kim Young Chul
Published: November 2023

Paraquat (PQ), a widely used herbicide, is highly toxic to both plants and animals. A toxic dose of PQ can result in severe lung injury in which oxidative stress is suggested to play a major role. Betaine is a methyl donor utilized in remethylation of homocysteine to methionine, which is mediated by betaine‐homocysteine methyltransferase (BHMT) mostly localized in the liver. However, our recent study revealed that the metabolism of sulfur‐containing substances in extrahepatic tissues was also influenced significantly by betaine intake, probably via a change in supply of sulfur‐containing amino acids in blood. In this study, we determined the effect of betaine against PQ‐induced oxidative stress and pulmonary fibrosis in rats. Rats were instilled with PQ (0.3 mg/kg) intratracheally. Betaine was provided in drinking water (1 %) from 2 weeks before the PQ challenge. In 2 weeks after PQ instillation, 4‐hydroxyproline levels in the lung and oxidative DNA damage measured by the Comet assay were increased significantly. Similar results were shown in histopathological assessment of the lung. Betaine supplementation effectively inhibited the fibrogenic changes in the rats treated with PQ. PQ instillation decreased methionine levels in the lung, but betaine supplementation elevated methionine and S‐adenosylmethione (SAM) levels significantly. Putrescine and spermidine levels were also elevated by betaine supplementation. On day 4 after PQ instillation, the metabolomics of sulfur‐containing substances was changed markedly, but glutathione (GSH) and its metabolic substrates, including methionine, SAM and cysteine, were all elevated in the lung by betaine supplementation. Taken together, the results suggest that betaine may protect the lung from PQ‐induced oxidative stress and pulmonary fibrosis most probably via enhancement of antioxidant capacity and polyamine synthesis.

Support or Funding Information

Supported by National Research Foundation (NRF) grants (No. 2014‐R1A2A1A11052967 and No. 2009‐0083533) funded by the Ministry of Education, Science and Technology (MEST), Korea.
Apoptosis‐Inducing Herbs for Prostate Cancer Prevention and Treatment
Authors: Meng Jing , Hawthorne Benjamin , Lund Kaleb
Published: November 2023

Background

Prostate cancer is the most common non‐skin cancer in men. The conventional treatment regimen for prostate cancer is chemical castration, which has adverse effects, and tumors often recur following treatment. Therefore, developing an alternative therapeutic plan for prostate cancer is very much desirable. The purpose of this study is to identify and validate natural herbs that can induce apoptosis of prostate cancer cells.

Methods

Saw Palmetto (Serenoa repens, I), Sanqi (Panax notoginsengI), and Ban Lan Gen (Isatis tinctoria) are extracted in alcohol at various concentrations. We performed HPLC to characterize these herbs, and then treated LNCaP prostate cancer cells with the ethanol extracts of these herbs. The viability of the treated and control cells was assessed by MTT Cell Proliferation Assay and flow cytometry. Real‐Time Quantitative PCR was performed to determine the expression levels of apoptotic genes.

Results

We show here that Saw Palmetto (Serenoa repens, I), Sanqi (Panax notoginsengI), and Ban Lan Gen (Isatis tinctoria) have the corresponding standard HPLC molecular profiles. They decreased the cell viability and enhanced apoptosis of LNCaP cells based upon MTT assay and flow cytometric analysis. Our preliminary quantitative RT‐PCR data also showed the elevated the expression levels of apoptotic genes in LNCaP cells treated by the herb extracts.

Discussion

The data show that cell proliferation of prostate cancer cells is reduced by treatment in vitro of the herbs studied. Despite the limitations of this in vitro study, these data provide exciting evidence for the anti‐cancer properties of these herbs and could contribute to new treatments for prostate cancer.

Support or Funding Information

This work was funded by Grant 4 and 13 from Bastyr University.
The Role of Aminoguanidine on the Epididymis of Diabetic Rats: A Histological Study
Authors: Othman Manal Ahmed
Published: November 2023

Background

Diabetes mellitus (DM) is a degenerative disease with alteration in carbohydrate homeostasis that harmfully affect the male reproductive function. One of the diabetic complications is the production of advanced glycation end products (AGEs) that generate reactive oxygen intermediates. Consequently this might be deleterious to reproductive organs and lead to the development of spermatogenic dysfunctions. Aminoguanidine, is an AGEs inhibitor and therefore it might ameliorate the diabetic complications. The epidydimis in an important structure in the male reproductive system, where maturation and storage of spermatozoa take place. The aim of this work was to demonstrate the effect of aminoguanidine treatment on the epididymis of adult diabetic male rats.

Materials and Methods

A total number of 18 male Sprague Dawley (SD) rats (average weight 200 gm) were divided into three groups of 6 animals each. Group I: was the control group. Group II: included the diabetic rats which were injected a single dose of 80 mg/kg (IP) streptozotocin (STZ) in sodium citrate. Group III: included rats with STZ‐induced DM (as in group II) that were orally administered aminoguanidine through a gastric tube at a dose of 4 mg/kg/day for 3 months from the third day after the induction of DM until the time of sacrifice. After sacrificing the animals, the epididymis was obtained from the three assigned groups and processed for light microscopic evaluation.

Results

In the diabetic animals, the epidydimal tubules showed variable degenerative changes in the form of decrease sperms in the tubules, with condensed nuclei in some epithelial cells. A notable finding was the presence of variable amount of white blood cells inside the lumen of the tubules which might indicate an inflammatory process. There were irregular increase in PAS staining as well as disrupted collagen content. On the other hand, diabetic rats treated with aminoguanidine showed improvement in their epididymal structure.

Conclusion

Diabetes mellitus resulted in degenerative changes in the epididymis of SD rats. These changes were enhanced by the administration of aminoguanidine which may be useful in the treatment of diabetic reproductive complications.
Dysregulated Inflammation in Response to Hypoxia‐Confounded Injury
Authors: Mayeux Jacques , Adesina Sherry , Caesar Christa , Naylor Petrina Barnett , Woodard‐Grice Alencia
Published: November 2023

Traumatic limb injury can cause extensive blood vessel damage that impairs oxygen and nutrient delivery to nearby tissues, resulting in a hypoxic state. The normal course of tissue repair involves a proper balance of pro‐ and anti‐inflammatory signaling; this balance is essential for tissue recovery as it aids in the destruction, containment or removal of pathogens and facilitates the repair of damaged tissue. However, in periods of extended hypoxia‐confounded injury, inflammation may become dysregulated. The mechanisms by which extended hypoxia may influence inflammatory cell activation during the acute recovery period are unknown. We therefore sought to create an in vitro wound model of sustained hypoxia to investigate inflammatory cell activation during this acute recovery period. We hypothesized that sustained hypoxia would result in greater immune cell activation over 72 hours compared to normoxia.

We completed several experiments utilizing human monocytes (U937 cells), human dermal fibroblasts (HDFs), and human umbilical vascular endothelial cells (HUVECs) exposed to normoxia (~21% O2) or hypoxia (~1% O2) for 24, 48, or 72 hours. We sought to assess the impact of sustained hypoxia on cell proliferation (scratch wound assay) and immune cell activation (macrophage adhesion; transmigration of macrophages; expression of cell surface markers CD11b and CD14) at each of the time points listed above.

We found that hypoxia impaired endothelial cell proliferation in a scratch wound assay, decreasing their ability to cover the gap area after 72 hours. We additionally found that macrophage transmigration was increased in response to hypoxia and that this result was correlated with increased immune cell proliferation as evidenced by increased cell surface expression of CD11b and CD14. We believe these results will provide valuable insight into the effect of sustained hypoxia on inflammatory signaling, and perhaps more importantly, the time course of signaling changes following hypoxia‐confounded injury.
12‐HETE Signaling Through the G Protein‐Coupled Receptor 31 (GPR31) Regulates Pancreas Development and Disease
Authors: Anderson Ryan Mallory , Hernandez‐Perez Marimar , Tersey Sarah , Samala Niharika , Maier Bernhard , Mirmira Raghavendra G.
Published: November 2023

Inflammation is essential for pathogen defense and tissue repair, but drives many pathologies when dysregulated. For instance, inflammation plays a critical role in pancreatic β cell dysfunction, and is a hallmark of both type 1 and type 2 diabetes. Thus, the molecular pathways initiating and escalating islet inflammation are prime targets for novel diabetes therapeutics. One such inflammatory mediator, 12‐hydroxyeicosatetraenoic acid (12‐HETE), is an oxygenated derivative of arachidonic acid generated by 12‐Lipoxygenase (12‐LO). Pancreatic deletion of 12‐LO protects obese high fat diet‐fed mice from glucose intolerance, and improves insulin secretion in cytokine‐treated islets in a 12‐HETE‐dependent manner. In vitro assays have shown that 12‐HETE can ligate the G protein coupled receptor GPR31, but little is known about the activation of this pathway or its role in islet biology; we hypothesized that the pancreatic actions of 12‐HETE are mediated in part by GPR31. Accordingly, when the beta cell line βTC3 was treated with 12‐HETE, we measured an induction of ER stress markers and suppression of oxidative stress pathway markers; these effects were blocked by shRNA knockdown of Gpr31, suggesting that 12‐HETE/GPR31 signaling is a node of convergence for stress pathways during inflammatory response in mouse β cells. To further clarify the in vivo roles of this pathway, we generated loss‐of function zebrafish models for both 12‐lipoxygenase (alox12) and gpr31 using morpholinos and CRISPR/Cas9. Loss of alox12 diminished 12‐HETE production and unexpectedly impaired pancreatic growth. Similarly, the loss of gpr31 resulted in a severe reduction of pancreas tissues via impaired differentiation. We propose that 12‐HETE signaling via GPR31 is an important paracrine and/or autocrine pathway involved in diabetes pathogenesis that is also essential for pancreas formation during development.

Support or Funding Information

This work is supported by NIDDK grant R01 DK105588 02 to RM
Endomucin is necessary for pathologic retinal neovascularization in vivo
Authors: D'Amore Patricia A. , Park‐Windhol Cindy A. , Saint‐Geniez Magali
Published: November 2023

Many ocular pathologies, including wet age‐related macular degeneration (AMD) and the retinopathy of prematurity, are characterized by abnormal neovascularization. Endomucin‐1 (EMCN), is an endothelial cell‐specific, sialic‐rich glycoprotein. We have shown that EMCN is necessary for VEGF‐mediated proliferation, migration and tube formation by human retinal endothelial cells in vitro and for normal retinal vessel development in vivo. Motivated by these finding, we examined the role of EMCN in the formation of pathological vessels using murine oxygen‐induced retinopathy (OIR) and the laser‐induced choroidal neovascularization (CNV) models. Intravitreal delivery of siEMCN to P12 mice following exposure to hyperoxia led to a significant reduction in avascular area (14.2±2.6% vs 22.6±4.7%, P<0.05) and neovascularization (2.6± 0.3% vs 3.5% ± 0.5%, P<0.05) in mice with EMCN knockdown at P17 compared to siCtrl mice. In the laser‐induced model of CNV, comparison of fluorescein angiograms at day 14 indicated that the CNV lesions in siEMCN‐treated mice were smaller and significantly less leaky than in mice (32.2±1.4% vs. 40.6±4.1%, P<0.05). Our data indicate EMCN reduction/neutralization may be a potential therapy for treating pathologic ocular neovascularization.

Support or Funding Information

This work was supported by EY5318 and EY26539 (PD'A), a Knight's Templar fellowship (CPW) and P30EY003790.
Can Molecular Hippocampal Alterations Explain Behavioral Differences in Prenatally Stressed Rats?
Authors: Woolard Emily Alice , Miller Colette , McDaniel Katherine , Beasley Tracey , Oshiro Wendy , Moser Virginia
Published: November 2023

Studies in both humans and animals have shown that prenatal stress (PNS) can alter cognitive function and other neurological behaviors in adult offspring. One possible underlying mechanism for this may lie with alterations in hippocampal gene expression. The present study examined genotypical outcomes in adult male and female offspring of rats exposed to variable stress during pregnancy. Dams (n=12–13/treatment) were subjected to several non‐chemical stressors including intermittent noise, light, crowding, restraint, and altered circadian lighting, from gestational day (GD) 13 to 20. Tail blood was drawn on GD 12, 16 and 20 to verify a stress response. Corticosterone levels were not different between the stressed and non‐stressed dams on GD 12, but was significantly increased in stressed dams on GD 16 and 20 compared to controls. Dams gave birth on GD 22 (postnatal day or PND 0). Several behavioral tests were used to assess the cognitive and behavioral phenotype of the offspring from PND 49 through 86, including the Morris water maze and novel object recognition task. Stressed offspring showed increased reversal path length on the Morris water maze. This result indicates changes in the behavioral performance of offspring. On PND 91–92, offspring were euthanized and hippocampal tissue was collected. Genotypic outcomes of PNS were assessed by quantifying hippocampal mRNA expression of five select genes thought to be implicated in these behaviors. The select genes were Brain‐Derived Neurotrophic Factor (Bdnf), Tropomyosin Receptor Kinase 2 (Trk2), DNA Methyltransferase (Dnmt) 1, Dnmt3a, and Dnmt3b, with Hypoxanthine Guanine Phosphoribosyltransferase (Hprt) used as an endogenous control. PNS female offspring had significantly lower Dnmt3b expression than controls (p=0.02). A trend showed PNS female offspring had reduced Dnmt3a levels compared to controls, suggesting additional alterations in basal levels of hippocampal mRNA expression (p=0.06). These preliminary findings support a role for hippocampal gene expression in long‐term changes in offspring of prenatally stressed dams.

This is an abstract of a proposed presentation and does not reflect US EPA policy.
The Changes of the Learning and Memory Ability after Stimulation by STN‐DBS in Parkinson's Disease Rats
Authors: Hao Chunyan , Duan Hubin , Li Hao , Wang suping , Zhang Ce , Pan Lijuan
Published: November 2023

Objective

To investigate the changes of the learning and memory ability after stimulated by subthalamic nucleus deep brain stimulation (STN‐DBS) in Parkinson's Disease (PD) rats.

Methods

55 healthy male SD rats were randomly divided into normal control group (n=10), PD group (n=15), sham stimulation group (n=15) and stimulation group (n=15). Stimulation and sham stimulation groups were planted in unilateral STN‐DBS. STN‐DBS in stimulation group was powered on and was stimulated 7 consecutive days at the regular time, while Sham stimulation group complied with the same program except power on the STN‐DBS. The apomorphine rotation was observed at pre‐stimulation and post‐stimulation. The latency to find the platform, swimming speed, swimming distance and crossing times were detected with Morris water maze.

Results

The symptoms of PD rats were improved significantly after stimulated by STN‐DBS. Compared with PD model group and sham stimulation model group, not only escape latency of rats in the DBS stimulated model group was decreased (P < 0.01), but also swimming speed (P < 0.05)and swimming distance (P < 0.05) were descend. The crossing times、 swimming distance and swimming time in which the platform located of PD model group(p < 0.05) and sham stimulation model group(p < 0.05) were definitely lesser than stimulation model group.

Conclusion

STN‐DBS stimulation not only can improve the ability of cognition, but also delay the condition worsen..

Support or Funding Information

This study was supported by the National Natural Science Foundation Youth (Fund No.30600637), China Postdoctral Foundation (Fund No.2014M561207), Shanxi Scholarship Council of China(Fund No. 2011‐096 and Key projects No.4), The science and technology research projects of health department of Shanxi province (FundNo. 201301068), Shanxi Province Basic Research Program Youth Science and Technology Research Fund (Fund No.201601D011119).,
3D Human Platelet Lysate Gel Model: LPS and Poly (I:C) Increase Invasion of Human Umbilical Cord Mesenchymal Stem Cells
Authors: Jirakittisonthon Thitikan , Jimenez Jake , Abello Javier , Cromer Adrienne , Sun Susan , Weiss Mark
Published: November 2023

Mesenchymal Stromal Cells (MSCs) can promote wound healing and have ability to migrate into injured tissues. The in vitro methods commonly used to track the migration of cells are the scratch assay and the transwell migration assay. Here, the objectives are to develop a novel would model in 3 dimensions and to observe invasion responses of MSCs after stimulation by LPS or Poly(I:C). A 3D wound model was created by culturing MSCs suspended with in a platelet lysate gel. Next MSC growth and migration was evaluated by stimulation by LPS or Poly(I:C). Both LPS‐stimulated, a gram negative bacterial cell wall component, and Poly (I:C)‐stimulation, a representative of viral infection, produced more MSCs growth and invasion than control (3D gel alone). This study demonstrates that MSCs can grow and migrate within a novel 3D wound model and that MSCs respond to LPS and Poly I:C with enhanced growth and migration. This data suggests that MSCs respond with more rapid wound healing response following exposure to stimuli associated with infection.
Progressive aging changes in the optical properties of Rat brain
Authors: Shi Lingyan , Sriramoju Vidyasagar , Kaleem Asma , Dudley Raymond , Rodriguez‐Contreras Adrian , Ostos‐Valverde Aline , Alfano Robert R
Published: November 2023

The objective of this study to investigate how molecular changes caused by aging effects the absorption and fluorescence of cerebral cortex during the postnatal development in the rat. The aging dependent changes in the absorption and fluorescence properties of native chromophores in rat cerebral cortex were studied at postnatally from neonatal stage to adulthood at days 5,10, 15, and 45. The primary chromophores in brain are responsible for specific absorption and/or fluorescence emission are collagen, tryptophan, NADH, flavins and water. The evolution in the relative concentrations of these key chromophores in the rat brains progressively with increasing age is presented. Understanding changes in the optical properties with age will help in developing better understanding the underlying development of molecular components in the brain and for imaging strategies for methods like optical tomography and multiphoton brain imaging.