bluuebunny/crossref_metadata_2025 huggingface dataset:
Desensitization of Basal Cell Carcinoma to The Anti‐Tumoral Effect of Vitamin D: Role of REDD1
Authors: Khasawneh Rawia , Unnikrishnan Archana , Mahmoud Bassel , Hamzavi Iltefat , Heydari Ahmad R
Published: November 2023

The relationship between vitamin D and non‐melanoma skin cancer is not clear. Epidemiological studies are inconsistent and inconclusive, and mechanistic studies are lacking. Vitamin D is a seco‐steroid hormone, whose nuclear receptor, vitamin D receptor (VDR), is a nuclear transcription factor. It has been suggested that vitamin D possesses anti‐proliferative and pro‐differentiation effects by negatively regulating important signaling pathways. In particular, the VDR target gene ‐ regulated in development and DNA damage response 1 (REDD1) ‐ inhibits the mechanistic target of rapamycin (mTOR) pathway, thus impacting cellular growth. In response to DNA damage, REDD1 transcription is up‐regulated by elevated levels of tumor protein 53 (p53), resulting in inhibition of mTOR pathway. Subsequently, reduction of p53 results in feedback inhibition at the translation level.

This case‐control study aimed to explore the impact of vitamin D status on the onset and progression and possibly treatment of basal cell carcinoma (BCC). Three tissue samples were collected from 20 BCC patients (Cancer, Proximal, and Distal), and 6 cancer‐free individuals from southeast Michigan. BCC and its feeding cells seemed to up‐regulate vitamin D activation enzymes, VDR, and its co‐activators, thus proposing higher local activity of vitamin D. Despite the significantly increased protein levels of REDD1 in the cancer tissue, our data showed that VRD's ability to down‐regulate mTOR pathway through REDD1 was diminished. Interestingly, Vitamin D negative regulation of Hedgehog‐Gli pathways was also lost in the cancer tissue. A gain of function mutation in the Kirsten Rat Sarcoma Viral Oncogene Homolog (KRAS), upstream of mTOR pathway, was detected in all BCC specimens. In conclusion, we propose that the anticipated role of vitamin D is not conserved in BCC tissue, possibly related to additional mutations.
Stress Level Related to Dietary Intake and Metabolic Syndrome in Middle Aged Women in Urban Area
Authors: Ko Hyunmi , Byeon Suji , Shin Yoonjin , Kang Bori , Doo Miae , Lee Seohyun , Kim Yangha
Published: November 2023

The aim of this study was to investigate dietary intake and metabolic syndrome risk based on the level of stress in middle‐aged woman in urban area. Subject were participants from the Health Examinees (HEXA) study of the Korean Genome and Epidemiology Study (KoGES). A total number of 81,639 middle‐aged women were classified into low stress group (n=47,395) and high stress group (n=342,44) according to their score of Psychosocial Well‐being Index‐Short Form (PWI‐SF). Data was collected using self‐administered questionnaires, anthropometric measurements and blood chemical analysis. High stress group showed significantly higher BMI, waist circumference, and hip circumference than low stress group. Moreover, subject with high stress showed higher fasting blood sugar and lower HbA1c and HDL‐cholesterol than low stress group. High stress group showed higher intake of carbohydrate and lower intake of the other nutrients than low stress group. In the high stress group, odds ratio of metabolic syndrome and related risk factors, such as abdominal obesity, elevated triglyceride, and elevated fasting blood sugar levels, were significantly increased than low stress group. Moreover, the higher carbohydrate intake showed the greater odds ratio of metabolic syndrome. The higher intake of protein, fat, retinol, vitamin E, and vitamin C showed the lower odds ratio of metabolic syndrome. Thus, the present study suggested that high stress might be associated with high intake of carbohydrates and low intake of the other nutrients including antioxidant vitamins, which could contribute to increased risk of metabolic syndrome in middle‐aged woman in urban area.
The Influence of Dietary Supplementation of Bacteriophages on Energy and Nutrient Digestibility and Intestinal Microbiota of Pigs
Authors: Jo Hyunwoong , Kim Eun Bae , Han Geon Goo , Kim Beob Gyun
Published: November 2023

The aim of this work was to measure the influence of supplemental bacteriophages (BP) on the microbiota in the small and large intestine and energy and nutrient digestibility in growing pigs. Twelve barrows with a mean body weight of 30 kg fitted with a T‐cannula at the end of ileum were individually housed in pens equipped with a feeder and a nipple drinker. A quadraplicated 3 × 2 incomplete Latin square design with 3 dietary treatments, 2 periods, and 12 animals was employed to obtain 8 observations for each treatment. The 3 experimental diets were a corn‐soybean meal‐based control diet with no antibiotic or BP, a diet supplemented with 0.1% antibiotic, and a diet supplemented with 0.2% BP. Fecal samples were collected on d 5, and ileal samples were collected on d 6 and 7 of each period. Bacterial genomic DNA were extracted from the fecal and ileal digesta samples and the V4 regions of 16S rRNA gene were amplified. Amplicons were sequenced on Illumina MiSeq, and microbial communities were analyzed by using QIIME. The ileal digestibility of crude protein tended to be less in the BP group compared with the control group (81.0% vs. 83.8%, P = 0.08), but the total tract digestibility of crude protein was not affected by supplemental antibiotic or BP. The ileal and total tract digestibility values of energy, dry matter, organic matter, neutral detergent fiber, and acid detergent fiber were unaffected by supplemental antibiotic or BP. At the phylum level, pigs fed the BP diet tended to have a greater proportion of Firmicutes (68.8% vs. 60.5%, P = 0.092) and a less proportion of Bacteroidetes (11.9% vs. 18.4%, P = 0.081) in the ileal digesta compared with those fed the control diet. At the genus level, pigs fed the BP diet tended to have a greater proportion of Lactobacillus (38.4% vs. 28.0%, P = 0.072) and a less proportion of Bacteroides (2.4% vs. 4.6%, P = 0.068) in the ileal digesta compared with the control group. In the fecal samples, dietary supplementation of antibiotic or BP resulted in a reduced proportion of Bifidobacterium compared with the control group (1.6% or 2.0% vs. 3.5%, P < 0.10). In conclusion, dietary supplementation of bacteriophages positively affected the microbiota of the small intestine without largely affecting energy and nutrient digestibility in growing pigs.

Support or Funding Information

This work was supported by CTC Bio Incorporation and National Research Foundation of Korea.
Association Between Toenail Magnesium and Type 2 Diabetes in Chinese Adults
Authors: Zhang Jiguo , Ding Gangqiang , Wang Huijun , Wang Zhihong , Su Chang , Zhang Ji , Zhang Bing
Published: November 2023

Previous studies have showed an inverse association between magnesium level and type 2 diabetes, but the results are inconsistent, and the evidence relates only to dietary and serum magnesium. Moreover, it is not certain how these findings are applicable to Asian people. This study was designed to examine the association between toenail magnesium and type 2 diabetes in Chinese adults. The sample was 5,683 adults 18 years or older from the 2009 China Health and Nutrition Survey. We used hemoglobin A1c or greater than 6.5% as the diagnostic criterion for type 2 diabetes. Inductively coupled plasma–mass spectrometry determined toenail magnesium. Mean toenail magnesium in participants with and without type 2 diabetes was 263.0 ± 170.9 and 282.3 ± 191.9 micrograms per gram, respectively. The multivariable‐adjusted odds ratio for type 2 diabetes comparing the highest to the lowest quartile of toenail magnesium was 0.72 (95% confidence interval, 0.52–0.99). We found a statistically significant interaction between toenail magnesium and geographic region on the prevalence of type 2 diabetes (P for interaction = 0.0086). Our findings suggest that toenail magnesium is inversely associated with the prevalence of type 2 diabetes. Promoting intake of magnesium‐rich foods may bring considerable benefits for prevention of type 2 diabetes, especially in those at high risk.

Support or Funding Information

This research uses data from the CHNS. The CHNS receives financial support from the Carolina Population Center (5 R24 HD050924), University of North Carolina at Chapel Hill; the National Institutes of Health (NIH) (R01‐HD30880, DK056350, and R01‐HD38700); and the Fogarty International Center, NIH (5D43TW007709 and 5D43TW009077) for data collection and analysis of files from 1989 to 2011 and future surveys; and the China‐Japan Friendship Hospital, Ministry of Health, for CHNS 2009.
Nutriet Composition of Complementary Food Formulated From Millet, Maize, Plantain and Soybean
Authors: Adeoso Abiodun Olaitan , Oyebamiji Olufunke Layo
Published: November 2023

Formulation of complementary diets is an alternative to ready‐made canned foods from local available staples to cut cost and improve nutritional status of the Nigerian child. Dried ground flours from plantain and processed soybean were mixed with either roasted ground millet or maize flours to formulate two Blends‐A (maize only) and B‐(millet only) both at ratio 10: 30: 60. Another blend mixed at ratio 10: 30:30:30 named C from the four composite flours were also prepared. The blends were prepared into porridges for sensory evaluation thereafter. The crude fibre contents of processed soya bean, millet, maize and plantain were 4.75%, 8.5%, 6.0%, and 8.75% respectively. Ash content was 2.37%, 1.13%, 0.80% and 2.5% respectively following the sequence above. Crude protein in soybean was highest with 9.50%, millet 8.55%, maize 8.50% and plantain 4.50%. Crude fat was highest in soybean with 25.13%, 7.0% in millet, 7.5% in plantain and lowest in maize 5.75%. Six of the ten students who evaluated the porridges made from the three blends found blend B most acceptable. This they attributed to the malty taste of blend B which has a higher volume of millet because of its unusual taste, thickness and color. The blends can therefore be used as complementary foods to improve the nutritional status of children and as a substitute for common traditional pap made solely from maize.

INTRODUCTION

Protein‐Energy‐Malnutrition is one of the major threat to child development globally. In Nigeria, according to the 10 years retrospective review carried at the University of Nigeria Teaching Hospital Enugu, 1 out of every 35 children suffer from malnutrition. This unfortunate trend in a land flowing with milk and honey like ours is due to ignorance mainly as so many mothers believe that only “canned” foods can do the infant good. Although many works has been done on complementary foods, but the general acceptability is low till date

OBJECTIVE

The quest to use locally produced staples in the production of complementary foods that are cheaper than the commercially available ones and at the same time nutritious gave birth to this work.

COMPOSITE FLOUR FORMULATION

Three blends of composite were formulated as follows.

Plantain, soybean and maize (10:30:60). (Blend A)

Plantain, soybean and millet (10:30:60). (Blend B)

Plantain, soybean and maize and millet (10:30:30:30). (Blend C)
Ensuring All Infants Have Access to Human Milk: A Systematic Approach for Establishing Sustainable and Integrated Human Milk Banking Programs
Authors: Amundson Kimberly , Sachdeva Ruchika , Quynh Nga Nguyen , Muiruri Juliana , Israel‐Ballard Kiersten
Published: November 2023

Background

Human milk is a pillar for infant survival, and when mother's own milk is unavailable or insufficient for low‐birth, preterm infants donor human milk processed and screened by a human milk bank (HMB) is the next best option; yet guidance for the introduction of HMBs as a critical component within breastfeeding promotion programming is especially limited for low and middle income countries.

Objective

To ensure sustainability by systematically strengthening systems for establishing HMB programs as an integrated approach so all infants have access to quality and safe human milk.

Methods

We developed a systematic approach for the introduction of integrated human milk banking globally, specifically in low and middle income settings. The first step involves establishing ownership through conducting formative assessments, establishing learning exchange opportunities with other HMB programs globally, hosting stakeholder meetings, and developing country‐specific strategy and guidance integrated into maternal and newborn care practices. The second step involves operationalizing an adapted system through a pilot introduction of an integrated HMB with quality control systems, and breastfeeding promotion and communications activities, all tailored for the local context. The third step involves evaluation of impact and cost of the implemented model for informing future implementation at scale. At each of these steps, engagement of local policy makers is essential for ensuring ownership and effective adoption into the health system.

Results

By building local technical capacity, operationalizing HMBs locally, and evaluating systems, local ownership is ensured for the prioritization of human milk for all infants. This methodology for strengthening HMB systems has been implemented in South Africa, and is ongoing in India, Vietnam, and Kenya. In each setting local governments have demonstrated ownership, working to adopt HMBs into the health system.

Conclusions

Where the burden of preterm and low‐birthweight are most prevalent, as is the case in low and middle‐income countries, integrated HMB systems, supported by the government, are essential for ensuring that all infants receive human milk. Further guidance and tools adaptable to setting‐specific needs is needed for implementation, integration, and regulation, and ultimate scale‐up of HMB programs globally to ensure that all infants have access to life‐saving human milk.

Support or Funding Information

The Bill and Melinda Gates Foundation financially supported the human milk banking implementation work in South Africa and India. Department for International Development, United Kingdom, through the County Innovation Challenge Fund, is financially supporting the human milk banking work in Kenya. The Margaret A. Cargill Foundation is financially supporting the human milk banking work in Vietnam and India.
Rationale and design of theory‐based nutritional strategy to promote healthy dietary habits in severe obese adolescents: Study protocol for ICAAN (Intervention for childhood and adolescent obesity via activity and nutrition) project
Authors: Kim Ji Eun , Jang Han Byul , Lee Hye‐Ja , Park Sang Ick , Kim YoonMyung , Seo Young‐gyun , Park Kyung‐Hee , Lim Hyunjung
Published: November 2023

Background

Behavioral modification on healthy eating and lifestyle is required to improve metabolically healthy status in severe obese child and adolescents. We aimed to develop tailored nutritional education programs to find barriers on dietary and lifestyle for severe obese adolescents and their caregivers.

Methods

Focused on theoretical framework approaches are effective to change behavior and achieve goals. To develop four monthly contents, social cognitive theory and transtheoretical model were applied to this program. All sessions were designed to give nutritional knowledge and practical mission in a real life for individual goal setting and self‐control. In the 1st and 2nd sessions, my meal planning and portion control were emphasized based on five food groups for diet and health outcome expectations. The 3rd and 4th sessions were build up skills to choose healthy snacks and cook with fruit and vegetables. 103 participants aged 8 to 16 years will be participated in a 16 week intervention program. During the each sessions, the participants and their family will be interviewed by nutritional expert for further tailored nutritional education and counselling.

Results

Each 20‐min individual face to face nutritional education and counselling will be offered every month for the participants and their parents; monthly phone contact and txt message for social support and online food recipe to emphasize having breakfast and vegetable and fruit consumption. Following as the monthly contents, individual dietary goal setting will be attained by every participants with their family. Promoting healthy eating practices resources; an educational book, practice posters, food diaries and a water bottle will be provided to all intervention participants to motivate healthy lifestyle behaviors.

Discussion

This nutritional intervention program might be assist to implement further researches in severe obese adolescents' population.

Support or Funding Information

Preparation of this article was supported by Grant 2015‐ER6401‐00 from the Division of Metabolic disease, Center for Biomedical science, Korea National Institute of Health.
Effect of Cirsium setidens Nakai extract on high‐fat diet‐induced obesity in C57BL/6J mice
Authors: Cho Bong‐Yeon , Lee Jin‐Ha , Kim Jae‐Min , Choi Sun‐Il , Jung Tae‐Dong , Choi Seung‐Hyun , Ra Moon‐Jin , Kim Sun‐Young , Kang Il‐Jun , Han Kyoung Chan , Lee Ok‐Hwan
Published: November 2023

Recently, a number of anti‐obesity drugs have been developed and used to treat obese patients through regulate appetite, fat absorption and fat oxidation. However, some drugs have undesirable side effects such as cardiac arrhythmia, constipation steatorrhea. Therefore, the demand for more effective and safe new anti‐obesity functional materials is still growing. Cirsium setidens, a wild perennial herb, has been widely used in korean medicine for treatment of hemorrhage, hypertension, and diabetes. In this study, we investigated the beneficial effect of Cirsium setidens Nakai extract (CNE) on high‐fat diet (HFD)‐induced obesity in C57BL/6J mice. C57BL/6J mice were fed a high‐fat diet to induce obesity and were orally administered CNE (25, 50, 100, 200 mg/kg) or Garcinia cambogia extract (GCE) (positive control, 100 mg/kg) for 8 weeks. CNE‐treated groups were reduced body weight and adipose tissue weight and improved serum lipid profiles through downregulation of PPARγ, C/EBPα, FABP4, SREBP‐1c and FAS and upregulation of adiponectin and CPT‐1 in obese C57BL/6J mice fed a high‐fat diet. These results suggested that CNE may have an anti‐obesity effect through the regulation of transcription factors related to adipogenesis, lipogenesis, and lipolysis.
Energy intake fluctuations during Muslim holiday season
Authors: Tutumlu Hazal , Goktas Zeynep
Published: November 2023

Introduction

Recent studies demonstrated that majority of annual weight gain occurs during short periods of time such as holidays. Assessing the critical time periods that predominantly contribute to annual weight gain could provide essential information to fight against obesity. There are some studies evaluating the Christian holiday season but there is not enough data for showing the effect of Muslim holiday season on body weight.

Objective

The purpose of this study to assess the energy intake fluctuations during Muslim holiday season of sedentary and physically active adults.

Methods

A total of 63 adults aged between 19 to 64 years participated in the study. Subjects were evaluated 5 times at following periods; before Ramadan (May 2016), during Ramadan, during Eid Al Fitr, after Ramadan and during Eid Al Adha (September 2016). Anthropometric measurements were collected using bioelectrical impedance analysis. During these measurements subjects were instructed to be fasted for at least 4 hours prior to testing. Nutritional evaluation carried out using 3‐day food records. Twenty‐four hour physical activity records are also collected to assess physical activity level. Statistical significance was set at p<0.05.

Results

A total of 39 female and 24 male subjects participated in the study. 47.6% of the subjects were physically active and carrying out regular exercise at least 3 days in a week whereas 52.4% of the subjects were sedentary. For the sedentary group, energy intake levels showed a significant increase from before Ramadan to during Ramadan (p < 0.05), whereas physical active group showed no change during this time period. Both groups showed an increase in energy intake during Eid Al Fitr and both groups showed a decrease in energy intake after Ramadan (p < 0.05). This decrease was followed by a significant energy intake increase during Eid Al Adha for both groups (p < 0.05). Energy intake levels were significantly increased between the baseline visit and the last visit for both groups (p < 0.001).

Conclusion

Both sedentary and physically active adults seem to increase energy intake during Muslim holiday season.
Spontaneous differentiation and organotypic tissue formation in 2D and 3D cultures of bovine mammary ductal stem‐like cells isolated directly from fresh milk samples
Authors: Lyden Timothy , Kehoe Sylvia , Plautz Emilee , Archambault Megan
Published: November 2023

Following up on literature reports of the harvest and culture of stem cells from normal human breast milk, our group at the UWRF Tissue and Cellular innovation Center has focused an ongoing study on extending those earlier reports using bovine samples. And further, to apply the resulting monolayers to 3D hanging drop (HD) spheroid cultures in order to explore the formation of organotypic artificial ductal tissues. To date, we have successfully harvested, cultured and analyzed more than 8 samples from each of two cows in our University Farm herd. Samples were harvested for culture and flow cytometric analysis. During culture, cell morphology and behaviors were observed and documented and samples prepared for further labeling studies to confirm our flow cytometric data. Analysis of 2D cultures demonstrates that a distinctive stem‐like cell population was indeed isolated from each sample, although distinct differences in absolute cell yields were seen between cows and between multiple samples from the same cow. Isolated stem‐like cells were shown to have an overall dendritic appearance and displayed significant motility until arrival in the vicinity of differentiated epithelial cells, at which point these cells rapidly settled and laid down in proximity to the epithelial cells. Over the following 24 hours these stem‐like cells divided and underwent differentiation to increase the adjacent patches of epithelial tissue. Over a two month culture period, all samples retained a significant population of these motile, dendritic cells, suggesting that they actively divide in the culture conditions provided. Subsequent mammary ductal differentiation was observed in all the 2D monolayers formed. These included development of distinct regions containing lactocytes and evidence of milk fat generation, with layers of fat droplets floating in all wells with monolayers. Current efforts are now focused on the generation of hanging drop artificial tissues from our existing monolayer cultures which contain a combination of stem‐like and differentiated cells. Additional flow cytometry studies will also be performed on these monolayers to confirm the distribution of cells loaded into each HD well.
Targeting the Protein‐Protein Interaction between Gαh and PLCδ1 Prevents Metastatic Progression in Triple‐Negative Breast Cancer
Authors: Lin Yuan‐Feng , Huang Shang‐Pen , Chen Chi‐Long
Published: November 2023

The distant metastasis of triple‐negative breast cancer (TNBC) to other organs, e.g. lung, has been correlated with poor survival rates in breast cancer patients. Therefore, the identification of useful drug‐targets to prevent metastasis or even inhibit tumor growth of TNBC is urgently needed. Here we showed that the increased level of cytosol, not extracellular, Gah is a poor prognostic marker in breast cancer patients and correlates with the metastatic evolution in TNBC cells. Moreover, the G protein, not transamidating, activity of Gαh was appeared to be critical for promoting the metastatic potentials of TNBC cells. We further showed that the protein‐protein interaction (PPI) between Gαh and PLCδ1 plays a pivotal role in regulating cellular invasion ability of TNBC cells and the signature of combining high Gαh/PLCδ1 levels refers to a worse prognosis in clinical patients with breast cancer and correlates with lymph node metastasis of ER(−) breast cancer. Our data revealed that blocking the PPI of Gαh/PLCδ1 complex by the synthetically myristoylated PLCδ1 peptide corresponding to the Gαh‐binding interface appears to significantly suppress cellular invasiveness in vitro and inhibit in vivo lung‐metastatic colonies of TNBC cells. This report establishes Gαh/PLCδ1 as a poor prognosis factor in breast cancer patients and provides a therapeutic value of targeting the PPI of Gαh/PLCδ1 complex for combating the metastatic progression of TNBCs.

Support or Funding Information

This study was supported Ministry of Science and Technology (MOST 104‐2320‐B‐038‐061‐MY3) and Taipei Medical University (TMU103‐AE1‐B13) to Dr. Yuan‐Feng Lin.
NOS1 and SOC1 work as a switch of macrophage functional phenotypes
Authors: Blajszczak Chuck , He Chenxia , Gantner Benjamin N , Bonini Marcelo
Published: November 2023

Nitric oxide synthases are enzymes involved in the production of nitric oxide (NO) from L‐arginine. NO is an important molecule with critical roles in signaling including inflammatory signaling. We found that differently than other isoforms, NOS1 (neuronal NOS or nNOS) is unique in the sense it promotes inflammation. We have uncovered that NOS1 is actually essential to start and amplify inflammation. This is because we demonstrated that in contrast to the enhanced susceptibility of NOS2−/− and NOS3−/− mice to inflammatory stimuli (i.e. LPS), NOS1−/− mice are, in fact, resistant both in terms of lethality and tissue injury. We also show that NOS1 deficiency not only attenuates TLR4‐stimulated pro‐inflammatory cytokine production but it also switches cytokine profiles from pro‐ to anti‐inflammation in macrophages. We found that this is because NOS1 inhibits SOCS1 which normally degrades p65 inhibiting the master regulator of inflammation NF‐κB (p65/p50 heterodimer). Hence, the suppression of NOS1 activates SOCS1 leading to the degradation of NFkB p65/p50 (inflammatory configuration) and the accumulation of p50/p50 (anti‐inflammatory configuration). Moreover we show here that the transplant of NOS1−/− macrophages which assume an anti‐inflammatory phenotype corrects exaggerated inflammation in vivo. Mechanisms will be discussed.

Support or Funding Information

NA
Exploiting ROS‐Generating Mechanisms for IFN‐g Therapy of Renal Cell Carcinoma
Authors: Shahrokh Soroush , Makhov Peter B
Published: November 2023

Background

Renal Cell Carcinoma (RCC) is the most lethal urologic cancer and accounts for 2%–3% of adult malignancies. While significant advances in its treatment have been achieved in recent decades, metastatic RCC remains an essentially incurable disease. IFN‐g therapy initially showed promise in treating metastatic RCC, but the interest in its use subsided due to the resistance developed by the RCC cells against IFN‐g treatment and a lack of understanding about its mechanism of action. One major factor responsible for RCC resistance to IFN‐g treatment is Nuclear Factor Kappa B (NF‐kB), which has been shown to be activated by IFN‐g treatment. Therefore, the need for the use of safe NF‐κB inhibitors is deemed necessary along with the use of IFN‐g.

Purpose

To explore the anti‐tumor efficacy of IFN‐γ in combination with Piperlongumine (PL), a potent natural NF‐kB inhibitor, and elucidate the mechanism by which PL sensitizes RCC cells to IFN‐g mediated apoptosis.

Methods

To measure NF‐kB activity, we used Luciferase Reporter Assay. To measure gene expression of TNF‐Related Apoptosis Inducing Ligand (TRAIL) and Tumor Necrosis Factor‐a induced by IFN‐g and/or PL treatment, we used quantitative reverse transcription polymerase chain reaction (qRT‐PCR). To measure the level of cell apoptosis induced by IFN‐g and/or PL, we used Terminal deoxynucleotidyl transferase nick end labeling (TUNEL) assay. To determine the level of protein expression as the result of IFN‐g and/or PL treatment, we used Western Blotting.

Results

We found that IFN‐g mediates apoptosis through up‐regulating the expression of TRAIL and TNF‐a death ligands. Additionally, we found that PL mediates the suppression of NF‐kB activation though an elevation of the Reactive Oxygen Species (ROS) levels in RCC cells.

Conclusions

Our results indicate that PL limits RCC resistance to IFN‐g treatment, potentially opening a new therapeutic avenue for the treatment of metastatic RCC.
Differential Effects of sEH Inhibitors on Vascular Smooth Muscle Cell Proliferation and Migration
Authors: Kim Hyo Seon , Kang Keon Wook
Published: November 2023

Epoxyeicosatrienoic acid (EET), an arachidonic acid metabolite, is known to have anti‐inflammatory and cardioprotective effects. EET is metabolized by soluble epoxide hydrolase (sEH) into dihydroxyeicosatrienoic acid (DHET). Neointimal hyperplasia results from atypical proliferation and migration of vascular smooth muscle cells (VSMCs), which leads to vascular diseases including atherosclerosis and restenosis. Our study aim is to assess the potential role of sEH inhibitors on proliferation and migration of VSMCs.

sEH inhibitor AUDA (12‐[[(tricyclo[3.3.1.13,7]dec‐1‐ylamino)carbonyl]amino]‐dodecanoic acid) and CDU (NCND, N‐Cyclohexyl‐N′‐dodecylurea) inhibited platelet‐derived growth factor (PDGF)‐induced proliferation of rat VSMCs. Nrf2 activation and subsequent heme‐oxygenase‐1 (HO‐1) induction are reported to attenuate neointima formation. AUDA decreased Keap1 expression while increasing Nrf2 nuclear translocation in VSMCs. Also, in higher concentrations (above 10 μg/mL) AUDA induced HO‐1 expression. Our lab previously reported peptidyl/prolyl isomerase Pin1 upregulation in neointima lesions of the femoral artery. Pin1 overexpression inhibited HO‐1 expression as well as Nrf2/anti‐oxidant response element (ARE) pathway in VSMCs. In accordance with it, AUDA concentration‐dependently inhibited PDGF‐induced Pin1 expression. However, exogenous EET had little effect on HO‐1 and Pin1 expression or VSMC proliferation. Thus, sEH inhibitors regulate VSMC proliferation by HO‐1 induction and Pin1 suppression, which is possibly irrelevant to EET stabilization.

On the other hand, AUDA increased PDGF‐induced migration of VSMCs. Cyclooxygenase‐2 (COX‐2) metabolizes arachidonic acid to thromboxanes and prostaglandins, some of which are known to promote VSMC migration. sEH inhibitors AUDA and CDU increased PDGF‐induced COX‐2 expression in a concentration‐dependent manner. Further, AUDA increased PDGF‐induced prostaglandin E2 (PGE2) secretion, which was attenuated by co‐treatment with COX‐2 selective inhibitor celecoxib. Also, celecoxib significantly inhibited the enhancement of PDGF‐induced VSMC migration by AUDA. However, celecoxib did not alter the effect of AUDA on PDGF‐induced proliferation, indicating COX‐2 specifically induces VSMC migration. When treated with AUDA, PGE2 receptor EP antagonists AH6809 and L‐161,982 did not affect PDGF‐induced VSMC migration, while thromboxane A2 (TXA2) receptor TP antagonist ICI192,605 and PGI2 receptor IP antagonist CAY10441 significantly suppressed it. However, CAY10441 reduced PDGF‐induced VSMC migration even without AUDA, implying its effect might be nonspecific. Therefore, enhanced PDGF‐induced migration by AUDA may be attributable to COX‐2‐meidated TXA2 formation.

In conclusion, sEH inhibitors have differential effects on PDGF‐induced VSMC proliferation and migration; inhibition of PDGF‐induced VSMC proliferation via Pin1 inhibition and potentiation of PDGF‐induced VSMC migration via COX‐2 induction and subsequent TXA2 production.
The Atypical Antidepressants Mianserin and Mirtazapine Inhibit TNF‐α‐Induced Apoptosis via LPA1 in Mouse HT22 Hippocampal Cells.
Authors: Olianas Maria C , Dedoni Simona , Onali Pierluigi
Published: November 2023

Several studies have proposed that pro‐inflammatory cytokines, such as TNF‐α, contribute to the pathophysiology of major depression through different mode of actions, including direct impairment of hippocampal neural progenitor cell development and survival. However, little is known on the mechanisms by which drugs effective in treating major depression may counteract the deleterious effect of pro‐inflammatory cytokines on neural cells. We have recently reported that different antidepressants activate the lysophosphatidic acid receptor LPA1 to induce growth factor receptor transactivation, cell proliferation and neuroprotection [1,2]. In the present study we show that in mouse HT22 hippocampal cells, which display a neuroblast‐like phenotype, the atypical antidepressants mianserin and mirtazapine inhibit the apoptotic cell death induced by TNF‐α and that this protective action involves the activation of LPA1. Thus, prolonged exposure of HT22 cells to TNF‐α increased caspases 8, 7 and 3 activities, proteolytic cleavage of poly ADP‐ribose polymerase and DNA fragmentation. Co‐treatment with antidepressants markedly attenuated the activation of the apoptotic cascade induced by TNF‐α. Blockade of the LPA1 receptor with the selective antagonists AM966 and Ro6842262 antagonized the anti‐apoptotic activity of the antidepressants. Analysis of intracellular signalling showed that through activation of LPA1 antidepressants transactivated the fibroblast growth factor receptor and triggered the phosphorylation of ERK1/2 and CREB, two key regulators of neural survival and differentiation. Collectively, the data indicate that in HT22 hippocampal cells LPA1 is a critical mediator of the neuroprotective effects of mianserin and mirtazapine against the pro‐apoptotic action exerted by TNF‐α.

Support or Funding Information

Supported by University of Cagliari. Italy
Acetate, the Metabolite of Ethanol, Increases Cytosolic Calcium and mRNA Expression Levels of EGR1 and TNFα in Dopaminergic Like PC12 Cells
Authors: Behnke Jessica E. , Chapp Andrew D. , Driscoll Kyle M. , Shan Zhiying , Chen Qinghui
Published: November 2023

We have previously reported that acetate activates NMDAR in vivo and in vitro, increases pro‐inflammatory cytokine mRNA expression, and cytosolic calcium levels, all of which may contribute to neurotoxicity. Neural growth factor (NGF) derived PC12 cells are a reasonable model used in substances of abuse and neurodegeneration studies as they are primarily dopaminergic in nature. Neurotransmitter release is governed in part from rises in cytosolic calcium levels which facilitate loading of synaptic vesicles and exocytosis from the synaptic cleft. We hypothesized that acetate may have similar effects in NGF derived PC12 cells which may facilitate increased dopamine release contributing to alcohol dependence. NGF derived PC12 cells exposed to (2 mM) acetate had a significant (p<0.05) increase in cytosolic calcium relative to baseline, measured in real‐time with a fluorescent calcium probe (Fluo‐4AM). Additionally, acetate (2 mM) significantly (p<0.05) upregulated mRNA expression levels of a calcium dependent protein, early growth response 1 (EGR1) at 15 and 30 minutes post acetate treatment, 5 and 6 fold respectively relative to control. EGR1 is an early response protein implicated to alterations in synaptic plasticity and also the immune response. Furthermore, 30 minute treatment with acetate also significantly (p<0.05) upregulated mRNA expression of tumor necrosis factor alpha (TNFα), 2 fold compared to control. This data suggests that physiological circulating levels of acetate (2 mM) following ethanol consumption and metabolism lead to increased cytosolic calcium levels, and mRNA expression levels of EGR1 and TNFα in dopaminergic like PC12 cells. These in combination may contribute to the development of alcohol dependence and alcohol associated neurotoxicity.

Support or Funding Information

(Funding support: AHA 16PRE27780121, Andrew Chapp, R15‐HL129213 Zhiying Shan; R15‐HL122952, Qinghui Chen).
Intermittent High Fat Diet Exposure Attenuates Acquisition and Maintenance of Alcohol Drinking in Rats
Authors: Vithlani Nisha , Smith Anna M , Dickenson Kanhiya , Davis Jon F , Sirohi Sunil
Published: November 2023

Alcohol use disorder (AUD) poses a significant socio‐economic burden to modern society and is a chronic relapsing disorder. Chronic alcohol consumption not only impairs the emotional states but also induces malnutrition. Neuroendocrine peptides, which are known to be regulators of nutritional status, can impact the reinforcing properties of alcohol. Therefore, altered neuropeptides as a result of a nutritionally impaired state may impact alcohol consumption in alcoholics. However, nutritional and dietary alternatives are often overlooked in the management of AUD. We have recently reported that six weeks of intermittent exposure to a nutritionally complete high‐fat diet (NCHFD) attenuates alcohol drinking in rats, which has important clinical implications in AUD treatment. At present, it is unclear if NCHFD pre‐exposure duration less than 6 weeks would be effective enough to impact alcohol drinking. In the present study, we assessed the impact of two weeks of intermittent NCHFD exposure on alcohol drinking in male, Long Evans rats. The rats (n=6/group) were grouped based on their body weight, food and water intake and were given intermittent access to NCHFD (24 hrs, twice a week) or normal chow (controls) for two weeks. All rats had ad libitum access to chow and water always and their daily food intake was recorded. Following two weeks of the intermittent NCHFD regimen, 20% ethanol drinking was evaluated using two‐bottle choice alcohol drinking model on the days following NCHFD exposure. A pattern of caloric overconsumption and under consumption was seen in the NCHFD exposed rats on NCHFD and non‐NCHFD days, respectively. Alcohol intake was significantly reduced in the NCHFD group of rats compared to the chow controls. Furthermore, not only preference and acquisition but also maintenance of alcohol intake was attenuated following intermittent NCHFD exposure. No between‐group differences in water or total fluid intakes were observed, suggesting that intermittent NCHFD exposure selectively impacted alcohol intake. As we begin to understand the impact of altering nutritional status on alcohol drinking behavior, the present data provide fundamental insights into the critical components for evaluating such interventions in the management of AUD.

Support or Funding Information

This publication was made possible by funding, in part, from the NIMHD‐RCMI grant number 5G12MD007595 from the National Institute on Minority Health and Health Disparities and the NIGMS‐BUILD grant number 8UL1GM118967 to SS. We thank the XULA vivarium support, especially Ms. Nannette Simoneaux.
Novel MicroScale Solutions for Biophysical Characterization in Drug Discovery and Development
Authors: Heffern Charles
Published: November 2023

Here we present 2 unique and proprietary technologies for biophysical characterization of biomolecules to facilitate the drug discovery and drug development process:

MicroScale Thermophoresis (MST) monitors the movement of molecules through μm‐sized temperature gradients to quantify interactions of any kind of molecule. Recent results from automated high‐throughput primary and secondary screening campaigns by MST will be presented. In a collaboration project with Sanofi R&D, MST identified several hits which were not detected by orthogonal techniques. In addition, MST detected secondary effects such as protein aggregation and denaturation, and could thus be used to reliably rule‐out false positives and measurement artifacts.

nanoDSF technology is specifically tailored for thermal and chemical unfolding studies. By monitoring fluorescence shifts of the amino acid tryptophan or tyrosine, unfolding profiles of multidomain proteins, transmembrane proteins or low‐abundant drug targets can be obtained in an entirely label‐free approach. Here, we present buffer and detergent screening campaigns for membrane proteins as well as for antibodies and antibody‐drug conjugates, highlighting the sensitivity and resolution of this method.

In addition, the fully‐automated protein stability platform using the novel Prometheus NT.Plex in combination with NT.Robotic Autosampler enables high‐throughput measurements as required in formulation development. Thermal and chemical unfolding data correlate with long‐term turbidity and monomer content over time, showing that the Prometheus NT.Plex can be used to rapidly predict the long‐term stability of biologics within 1 day.
Functional Improvement of Dystrophic Muscle via let7 Site Blocking Oligonucleotide Mediated Utrophin Upregulation
Authors: Mishra Manoj K , Loro Emanuele , Wilton Steve D , Khurana Tejvir S
Published: November 2023

Duchenne's muscular dystrophy (DMD) is a fatal genetic disease caused by the absence of dystrophin. Utrophin is the autosomal homolog of dystrophin and when overexpressed can compensate for dystrophin and rescue the dystrophic phenotype of the mdx mouse model of DMD. Utrophin is subject to miRNA mediated repression in muscle by several miRNAs including the let‐7c miRNA. Blocking the let‐7c‐utrophin interaction would be predicted to ‘repress the repression’ and increase utrophin expression. We developed and tested the ability of 2′‐O‐Methyl phosphorothioate (2OMePS)‐based utrophin 3′UTR let‐7c site blocking oligonucleotides (SBOs) to upregulate utrophin and improve the dystrophic phenotype in vivo. Blockade of the let‐7c‐utrophin interaction using bi‐weekly intraperitoneal injections of let7c SBOs for 1 month in young mdx mice led to increased utrophin expression along with improved muscle morphology, decreased muscle fibrosis and increased muscle specific strength. The functional improvement of dystrophic muscle achieved by using 2OMePS‐let7c SBOs suggests a novel, utrophin upregulation based therapeutic strategy for DMD.

Support or Funding Information

Acknowledgements: MDA, Shire Plc.
Pharmacokinetics of Cefquinome in Healthy and Pasteurella multocida‐Infected Rabbits
Authors: Elazab Sara T , Schrunk Dwayne , Griffith Ronald W , Ensley Steve M , Dell'Anna Giuseppe , Mullin Kathleen , Elsayed Mohamed G , Amer Magdy S , El‐Nabtity Sameh M , Hsu Walter H
Published: November 2023

The pharmacokinetics of cefquinome were studied in healthy and Pasteurella multocida‐infected rabbits after a single intramuscular injection at 2 mg/kg. 12 female New Zealand white rabbits (2.0–2.5 kg) were used; 6 of them served as controls, and the other 6 were infected; the experiments were done 1–2 days after Pasteurella nasal inoculation when rabbits showed signs of infection. Plasma concentrations of cefquinome were determined using high‐performance liquid chromatography. The values of elimination half‐life (T1/2 λz), area under the curve (AUC), area under the first moment curve (AUMC), and mean residence time were significantly lower in infected rabbits (0.47 h, 4.54 h*μg/mL, 3.63 h*h*μg/mL and 0.8 h) than healthy rabbits (0.72 h, 9.11 h*μg/mL, 9.85 h*h*μg/mL and 1.1 h), whereas total body clearance was significantly higher in infected than healthy rabbits. Therefore, P. multocida infection caused significant changes in some of the pharmacokinetic parameters of cefquinome in rabbits.

Support or Funding Information

Egyptian Ministry of Higher Education

Iowa State University supplied the tools and support to carry out the experiments.
Lindane (Gamma‐Hexachlorocyclohexane) Exposure Impairs [Ca2+]i‐Mediated Vascular Reactivity
Authors: Yakubu Momoh A , Abraham Nina S , Bessac Bret F
Published: November 2023

Gama hexacholorocyclohexane (γHCH; lindane), Gammallin is an organochlorine chemical that has been used as an agricultural insecticide as well as a pharmaceutical treatment for lice and scabies. It is reported to be neurotoxic, interfering with GABA neurotransmitter functions and other biological functions from brain to endocrine and cardiovascular systems. Despite being a banned persistent organic pollutant, it's still in use and can leach and contaminate our foods and water. We have investigated the effects and the mechanism(s) of lindane‐induced vascular dysfunctions in rat aorta. Rats treated with low dose of single pesticide Lindane (100 or 300 μmol/kg;1/5LD50; SP) or multiple pesticides (MP) (aldrin, endosulfan, lindane, 4,4‐DDT, and endrin: 1/100, 1/50, and 1/25 of the LD50, MP) orally or dermal application for 2 or 4 weeks showed significantly attenuated vascular relaxation (57 ± 5% to 36 ± 0.8%; ACh 10−3 M), MP attenuated relaxation (57 ± 5% to 19.8 ±5.8% and 19.0 ±4.0% respectively for 1/100 and 1/25 LD50). Paradoxically, 4 weeks SP enhanced relaxation to ACh (60.5 ± 4.6% to 83.5 ± 8.8%). In aorta from dermal group (SP), relaxation to ACh was significantly attenuated (59.4 ± 5.0 to 23.7 ± 6.0%). Vasoconstriction to PE (10−8 ‐10−3 M) were significantly attenuated by 4 weeks' treatment with oral SP or dermal MP reducing the contraction (0.64±0.06 to 0.15 ± 0.02gm or 0.49±0.01 gm; 10−3 M), but not for 2 weeks oral MP for PE‐induced contraction. Further investigation into the mechanism behind the observed vascular dysfunction reveals that in CHO cell line stably expressing M1‐ACh receptors, Lindane diminishes carbachol (10−8 to 10−3 M)‐induced increases in [Ca2+]i mediated by M1‐ACh receptor. Lindane diminishes Carbachol effects only at Carbachol's maximum activation and these results indicate that the vascular dysfunction by the pesticides could be due to disruption of intracellular Ca2+ regulation. Thus, 24‐hour Lindane exposure facilitates IP3‐mediated [Ca2+]i signaling. Month‐long Lindane exposure induces vascular dysfunction possibly via disruption of IP3‐mediated in [Ca2+]i signaling leading to dysregulated vasoresponsiveness to hormones/neurotransmitters contributing to cardiovascular dysfunctions.

Support or Funding Information

This research was supported by the National Heart, Lung, and Blood Institute (Grants HL03674 and HL070669) and by the use of TSU Research infrastructure support provided by grants G12RR003045 and CO6RR012537 awarded by the National Center for Research Resources, National Institutes of Health (NIH). The G12 program is now a part of the National Institute on Minority Health and Health Disparities (NIMHD) and the C06 program is in the Office of Research Infrastructure Programs in the Office of the Director, NIH.