bluuebunny/crossref_metadata_2025 huggingface dataset:
GRP78 is Not Expressed on the Surface of Triple Negative Human Breast Cancer Cells
Authors: Serrano‐Negron Jesus E , Romero Eva C , Sanchez Neysharie , Banerjee Aditi , Baksi Krishna , Banerjee Dipak K
Published: November 2023

GRP78, an Mr78 kDa glucose responsive protein is located in the lumen of the endoplasmic reticulum (ER). It functions as ER chaperone in translocating protein across the ER membrane that needs to be glycosylated at the asparagine residue present in the sequeon Asn‐X‐Ser/Thr (N‐linked glycosylation). This is an important biochemical event in protein folding. When N‐glycosylation is impaired with an N‐glycosylation inhibitor tunicamycin, angiogenesis, a hallmark for tumor progression and metastasis is inhibited. The result is regression of breast tumor. Reduced N‐glycan expression correlates with thinning and narrowing of the tumor microvessels. On the other hand, GRP78 expression in microvasculature and in tumor tissue is upregulated, concluding that GRP78 is a master regulator in ER stress induced unfolded protein response (upr)‐mediated apoptosis in tumor microvasculature (J. Biol. Chem. 286, 29127–29138, 2011; Pure Appl. Chem. 84, 1907–1918, 2012). This contradicts the current dogma that GRP78 expresses on the cell surface, interferes with the therapeutic approach(es) and becomes a tumor promoter. To evaluate the GRP78 localization in our study, we have used a triple negative (ER−/PR−/HER2−) human breast cancer cell line MDA‐MB‐231 cultured normally or without serum as well as before after inducing ER stress, and immunofluorescence microscopy. Unfixed cells were stained with Concanavalin A (Con A; Specificity = α‐D‐Mannose, α‐D‐Glucose, Branched mannose), Wheat germ agglutinin (WGA; Specificity = (GlcNAc‐β‐(1,4)‐GlcNAc)1‐4>β‐GlcNAc‐NeuAc) as well as with anti‐GRP78 antibody followed by their detection either with Rhodamin or Alexa–conjugated secondary antibody. The fluorescence images were captured in a Zeiss microscope with Axiocam camera. Con A and WGA staining have provided images of N‐glycans on the cell surface and supported the intactness of the cell membrane. On the other hand, GRP78 fluorescence was absent from the surface of the cancer cell. The results were the same whether the cells were cultured in the absence of serum and/or in the presence of 1μg/mL of tunicamycin. GRP78 fluorescence however was detectable in cells after either fixing them with ice‐cold methanol or permeabilizing them with digitonin. We, therefore, conclude that GRP78 is not expressed on the outer‐leaflet of the cell surface of the triple negative human breast cancer cells MDA‐MB‐231. But, its intracellular expression is anti‐tumorigenic.

Support or Funding Information

Supported in part by grants from NIH U54‐CA096297 and Susan G. Komen for Cure BCRT58206 (DKB) and NIH/NIMHD 2G12MD007583 (KB).
Plasma Kallistatin is Associated with Leukocyte Telomere Length in Young African Americans
Authors: Zhu Haidong , Chao Julie , Raed Anas , Huang Ying , Bhagatwala Jigar , Parikh Samip , Dong Yanbin
Published: November 2023

Objective

Large population studies demonstrate that African Americans versus their Caucasian counterparts have a steeper decline in leukocyte telomere lengths (LTL) with age. Telomere shortening plays an important role in cellular aging. Kallistatin, a novel tissue kallikrein inhibitor, has anti‐inflammatory and anti‐oxidant properties. Recently we have shown that kallistatin inhibited vascular injury and cell apoptosis in animal models and cultured cells. Thus we aimed to test our hypothesis that plasma kallistatin levels are positively associated with LTL in African Americans.

Methods

Leukocyte telomere lengths (the relative ratio of telomere repeat copy number (T) to single‐copy gene copy number (S)) were determined by a quantitative polymerase chain reaction method in 188 (60 males and 128 females) apparently healthy young African American subjects aged 13 to 49 years. Plasma kallistatin levels were measured by enzyme‐linked immunosorbent assay.

Results

Compared to males, females were older (24.0 ± 9.19 vs 17.9 ± 4.5 years, p<0.001), and had slightly shorter LTL (T/S ratio 1.00 ± 0.23 vs 1.09 ± 0.27, p= 0.03). In the whole sample, plasma kallistatin levels are positively and significantly associated with LTL (r=0.31, p<0.001). The significance persisted after adjusting for age, sex, and body mass index (r= 0.16, p=0.04).

Conclusion

This is the first study to examine the relationship between kallistatin and telomeres in humans. Kallistatin may have anti‐cellular aging effect, which may be attributed to its anti‐inflammatory and anti‐oxidant properties. These finding indicate that plasma kallistatin might serve as a biomarker for cellular aging in humans.

Support or Funding Information

HZ and YD are supported by NHLBI PO1 HL69999 and AHA 14GRANT20480211.
Adipose Depot‐Specific Differences in Transcriptome and MicroRNA Expression in High Fat Diet Induced Obese Mice
Authors: Wijayatunga Nadeeja N , Pahlavani Mandana , Kottapalli Rao , Dawson John A , Ramalingam Latha , Gunaratne Preethi H , Coarfa Cristian , Rajapakshe Kimal , Kalupahana Nishan S , Moustaid‐Moussa Naima
Published: November 2023

Obesity is characterized by white adipose tissue (WAT) expansion which contributes to chronic diseases such as diabetes and cardiovascular disease. Two main types of adipose tissue are WAT (subcutaneous, SAT and visceral adipose tissue, VAT) and brown adipose tissue (BAT). These exhibit anatomical, physiological and metabolic differences that contributes to energy balance, metabolic regulation and immunity. Our aim was to compare transcriptomic and microRNA (miRNA) signatures of SAT, VAT and BAT to gain mechanistic insight into the role of these depots in metabolic disorders. We performed RNA‐sequencing and miRNA profiling of SAT, VAT and BAT depots from diet‐induced obese C57BL/6J male mice fed high fat diets (45% kcal fat). We used a transcriptomic approach to dissect adipose depot‐specific genetic and functional signatures in obesity. RNA sequencing was performed using Illumina Hi‐Seq and differential gene expression was studied by pair wise comparisons of fat depots. Functional and pathway analyses performed using Ingenuity® Pathway Analysis software revealed inflammatory pathways such as cytokine and integrin signaling increased along with immune‐related processes in VAT compared to BAT. In contrast, pathways involved in fatty acid oxidation, apoptosis and energy expenditure (peroxisome proliferator activated receptor (Ppar)/retinoid X receptor (Rxr‐α)) were higher in BAT than in VAT. X‐box binding protein‐1 (Xbp1), an unfolded protein response (UPR) mediator in endoplasmic reticulum (ER) stress, was a common upstream regulator that was consistently activated in both VAT and SAT in comparison to BAT. Xbp‐1 targets include ER related genes such as glucose‐regulated protein 78 and 94 (Grp78, Grp94), calreticulin, protein disulfide isomerase family A 3 and 6 (Pdia3 and Pdia6) all of which showed higher expression in VAT than BAT. Interestingly, miR‐30c‐2‐3p which causes downregulation of Xbp1 was lower in VAT than in BAT, in agreement with higher Xbp1 levels in VAT compared to BAT. Furthermore, other miRNA‐mRNA pairs involved in UPR such as miR‐455‐calreticulin were also identified. Additionally, miRNAs miR‐221‐3p and miR‐222‐3p, related to higher inflammation and ER stress were activated in VAT compared to BAT, suggesting more inflammation and ER stress in VAT compared to BAT as expected. In summary, high fat diets differentially regulate specific miRNAs and gene expression in different adipose tissue depots with significant induction of ER stress and inflammation in VAT. Currently, studies are ongoing to validate these findings which would provide insight into the contribution of different fat depots to various metabolic pathways and will potentially impact various diseases associated with obesity.

Support or Funding Information

Startup funds from TTU
Location of Iron and Ferritin in the Midgut of the yellow fever mosquito, Aedes aegypti
Authors: Love Maria B , Geiser Dawn L , Winzerling Joy J
Published: November 2023

Dengue and Yellow Fever are caused by viruses transmitted by mosquitoes that each year infect 390 million and 200,000 people, respectively, and cause more than 2 million deaths. For mosquito eggs to develop and produce viable offspring, iron is required as an essential nutrient from the blood meal. Our previous research has shown iron from the blood meal is absorbed and transported by ferritin from the midgut to the ovaries in Aedes aegypti (yellow fever mosquito). Thus, understanding iron accumulation and utilization in the mosquito midgut is important to finding potential strategies to interfere with mosquito fecundity, decrease mosquito populations, and reduce transmission rates of vector‐borne diseases. However, it is unknown where in the midgut iron accumulates after a blood meal. To demonstrate this, Ae. aegypti midguts were dissected from sugar fed, 24 h post‐blood meal (PBM), and 72 h PBM animals to measure tissue iron accumulation after a blood meal by iron inductively coupled plasma mass spectrometry (ICP‐MS). Dissected midguts were also stained with Prussian Blue to visualize ferric iron deposition. Our results indicate that iron is present in the midgut prior to a blood meal, accumulates at 24 h PBM, and decreases by 72 h PBM, and the deposition of ferric iron is observed throughout the tissue before and after a blood meal. Further, immunohistochemistry studies of the midguts show three different subunits of the iron storage and transport protein, ferritin. Coalescence of ferritin and iron in mosquito midguts confirms the role of ferritin in this tissue for export of iron to other tissues, including the ovaries for oogenesis and ovarian homeostasis.

Support or Funding Information

This research was supported by funds from the Arizona Agricultural Experiment Station, the College of Agriculture and Life Sciences, the Undergraduate Biology Research Program, and the Honors College at the University of Arizona.
Blueberry supplementation attenuates oxidative stress within the monocytes and modulates immune cell levels in adults with Metabolic Syndrome
Authors: Nair Anand R , Mariappan Nithya , Stull April J , Francis Joseph
Published: November 2023

Background and rationale

Blueberries (BB) have been shown to have extensive health benefits in animals, including improvement of mitochondrial integrity, inflammation and redox status. In addition, we have found that BB improves insulin sensitivity in obese, pre‐diabetic humans and rodents, and decreases oxidative stress and inflammation, and protects against cardiovascular and renal damage in rodents. These findings indicate that BB have a whole body effect and are not limited to a particular organ system. In order for BB to exert beneficial effects on the whole body, the mechanism would logically have to operate through modulation of cellular humoral factors. The blood leukocytes are important cells protecting against injury or infection. The present study investigated the role of BB in modulating cellular humoral factors in subjects with metabolic syndrome (MetS).

Method

A placebo‐controlled study was conducted on adults with MetS, where they received a BB or placebo smoothie twice daily for six weeks. Free radical production in the whole blood and monocytes, dendritic cells (DC) measurement, gene expression levels of cytokines in monocytes and serum inflammatory marker levels were assessed pre‐ and post‐intervention. MetS patients

Results

Baseline free radical levels in the whole blood and monocyte of MetS patient samples were not different in both groups.. In contrast, treatment with BB markedly decreased superoxide and total ROS in the whole blood and monocytes compared to placebo treated MetS patients samples (p<0.05). Next we examined the dendritic cell number and inflammatory markers in the blood of MetS patients. There was no significant difference in the baseline DC number for both myeloid and plasmatoid DCs in MetS patients samples. Treatment with blueberries markedly increased myeloid DC and had no effect on plasmatoid cells. In addition, BB treatment also decreased monocyte and plasma inflammatory molecules in MetS patients when compared with placebo treated controls. These findings indicate that BB exerts immuno‐modulatory effects, in addition to its antioxidant and anti‐inflammatory effects, thereby protecting against MetS.

Conclusions

Blueberries modulates immune cells and attenuate oxidative stress and inflammation in the blood of MetS patients.

Support or Funding Information

This work was supported by U.S. Highbush blueberry council (USHBC) grant to Dr. Francis.
A Novel Element in the Cytoplasmic domain of the Type I TGFβ Receptor Mediates Basolateral Targeting in Polarized Epithelial Cells
Authors: Yin Xueqian
Published: November 2023

Delivery of biomolecules to the correct subcellular locales is critical for proper physiological function. To that end, we have previously determined that the type I and II TGFβ receptors (TβRI and TβRII, respectively) localize to the basolateral domain in polarized epithelia. While TβRII targeting was shown to be regulated by sequences between amino acids 529–538, the analogous region(s) within TβRI is unknown. To address that question, sequential cytoplasmic TβRI truncations and point mutations identified a targeting motif between residues 158 to 163 (VxxEED) required for basolateral TβRI expression. Further studies documented that receptor internalization, downregulation, direct recycling, or Smad signaling were unaffected by motif mutations which caused TβRI mislocalization. However, a 70 amino acid stretch containing the targeting motif was able to direct basolateral expression of the apically sorted Neurotrophin receptor (p75; extracellular and transmembrane regions) in a dominant manner. Lastly, co‐expression of apically targeted type I and type II TGFβ receptors mediated Smad3 signaling from the apical membrane of polarized epithelial cells. These later findings demonstrate that the absence of apical TGFβ signaling in normal epithelia reflects domain‐specific receptor expression and not an inability to couple with the signaling machinery.

Support or Funding Information

This work was supported by Public Health Service grants GM‐055816 and GM‐054200 from National Institutes of General Medical Sciences.
CREB regulates the expression of PERK and IRE1a, and controls unfolded protein response under hypoxic conditions
Authors: Nakayama Koh
Published: November 2023

Cells are frequently exposed to different stresses. Hypoxic condition induces ER stress, and activates unfolded protein response (UPR) to maintain homeostasis. Whereas Hypoxia‐inducible factor plays a major role in an adaptation to hypoxia, we previously identified that transcription factor CREB also has an important role in this process. To further understand the role of CREB in hypoxic response, CREB stable knock‐down cells (CREB‐KD) were established from breast cancer MDA‐MB231 cells and analyzed.

CREB was found to be activated by ER stress, and the activation of CREB and UPR pathway occurred in a coordinated manner by various stimuli, including ER stress‐inducing chemicals, hypoxia and oxygen‐glucose deprivation (OGD). Depletion of CREB decreased the expression of PERK and IRE1a, two of the critical UPR signaling molecules. ChIP assay and promoter analysis indicated that CREB interacts with the promoter region of these molecules and regulate the expression of them. ER stress induced by hypoxia was found to be reduced in CREB‐KD cells, which led to less pulmonary metastasis in mice. Finally, OGD strongly activated UPR and induced cell death in control cells, whereas this activity was moderate in CREB‐KD cells, thus more resistant to cell death.

Taken together, the study demonstrates a previously unknown role of CREB as a regulator of ER stress in cells, which is required for a proper response to the stressful conditions, such as hypoxia.

Support or Funding Information

Grant‐in‐Aid for Scientific Research C, and Grant‐in‐Aid for Scientific Research on Innovative Areas(KAKENHI).
Yeast strains heterozygous for YTA12 are sensitive to the anti‐cancer compound Parthenolide
Authors: Hojnacki Rachael Elizabeth , Callahan Kevin P
Published: November 2023

Parthenolide (PTL) is a naturally occurring small molecule that is the active ingredient in herbal medications used as a remedy for inflammation and pain. PTL exhibits exceptional anti‐cancer and anti‐inflammatory properties, making it a prominent candidate for furtherstudies and drug development. Acute myeloid Leukemia (AML) is a cancer of the blood, characterized by an overproduction of abnormal white blood cells that accumulate in the bone marrow. When acute myeloid leukemia cells are treated with PTL, there is a significant decrease in the function of the AML cells without altering or effecting normal cells. This is in contrast to current treatment for AML that are toxic and harmful to not only the cancer cells, but normal cells in the body. In hopes of better understanding the mechanism of PTL, the Haploinsufficiency Profiling (HIP) Assay was performedon Saccharomyces cerevisiae (bakers yeast). The HIP Assay consists of a complete set of heterozygous deletion strains that are grown in the presence of a compound of interest, strains most sensitive to the compound often carry deletions in genes that encode the drug target. The HIP Assay was performed in the presence of PTL. Yeast lacking one copy of the YTA12 gene, showed increased sensitivity to PTL. YTA12 encodes for a protein that iscomponent of the inner membrane protease that plays an important role in the degradation of incorrectly folded proteins. The hypersensitivity of the strain to PTL has been confirmed and current work is focused on determining the effect PTL has on the levelof YTA12 mRNA and protein.
Reactive Oxygen Species‐Dependent PKB Activation Contributes to Insulin‐Like Growth Factor 1‐Induced Phosphorylation of Histone Deacetylase 5 in Vascular Smooth Muscle Cells
Authors: Pietruczuk Paulina , Srivastava Ashok K
Published: November 2023

Insulin‐like growth factor 1 (IGF‐1), a potent mitogenic and vasoactive factor, has been shown to play an important role in the development of cardiovascular diseases. This occurs through the generation of reactive oxygen species (ROS) as well as through the hyperactivation of mitogenic and growth promoting signaling pathways and the subsequent alteration in gene expression. Histone deacetylases (HDACs), by their ability to modify the acetylation status of lysine residues in histone and non‐histone proteins, regulate gene transcription. Recent studies have demonstrated that a heightened activation of HDACs, notably HDAC5, is associated with vascular disorders such as atherosclerosis. However, a role of IGF‐1 in HDAC activation by phosphorylation remains poorly characterized. Therefore, in the present studies, we examined the effect of IGF‐1 on the phosphorylation of HDAC5 in vascular smooth muscle cells (VSMCs) and identified the signalling pathways involved in this process. Treatment of A10 VSMCs with IGF‐1 enhanced the phosphorylation of HDAC5 at serine 498 in a time and dose‐dependent fashion. Pretreatment of cells with AG1024, a selective pharmacological inhibitor of IGF‐1 receptor, significantly inhibited IGF‐1‐induced HDAC5 phosphorylation in A10 VSMCs whereas AG1478, a selective inhibitor of epidermal growth factor receptor, did not have an inhibitory effect on the levels of phospho‐HDAC5. Pharmacological blockade of the MAPK pathway with PD98059 and UO126 had no effect on HDAC5 phosphorylation, however wortmannin and SC‐66, inhibitors of the PI3K/PKB pathways, almost completely attenuated IGF‐1‐induced HDAC5 phosphorylation. In addition, pretreatment of A10 VSMCs with Diphenyleneiodonium (DPI) and apocynin, two NAD(P)H oxidase inhibitors, as well as the antioxidant N‐Acetyl‐Cysteine (NAC), resulted in an attenuation of IGF‐1‐induced phosphorylation of HDAC5 as well as PKB. In summary, these data demonstrate that IGF‐1 induces the phosphorylation of HDAC5 in a ROS‐ and PKB‐dependent fashion in VSMCs.

Support or Funding Information

Supported by a grant from the Canadian Institutes of Health Research.
O‐GlcNAc Regulates p53 in Ovarian Cancer
Authors: Queiroz Rafaela Muniz , Chien Jeremy , Madan Rashna , Slawson Chad , Dias Wagner
Published: November 2023

O‐GlcNAcylation is a dynamic post‐translational modification (PTM) that consists of an addition of a single N‐acetylglucosamine sugar to serine and threonine residues in nuclear, cytoplasmic and mitochondrial proteins. The enzyme O‐linked β‐N‐acetylglucosamine transferase (OGT) adds the modification, while enzyme O‐GlcNAcase (OGA) removes the modification. O‐GlcNAc regulates enzymatic activity, localization, stability and protein‐protein interactions on a wide range of proteins such as transcription factors, structural proteins and kinases. In cancer, when comparing O‐GlcNAc levels in normal and tumor cells, tumor samples present higher O‐GlcNAcylation in vitro and in vivo. This aberrant O‐GlcNAc pattern contributes to changes in important signaling pathways and could contribute to the progression of the disease. Interestingly, the tumor suppressor p53 is modified by O‐GlcNAc. The transcription factor p53 regulates diverse cellular processes like cell survival and the cell cycle. Most solid tumors contain mutations in p53 leading to increased degradation or loss of p53 function, which promotes malignant tumor growth. Ovarian cancer has one of the highest frequencies of p53 mutation rates. The goal of this work is to understand the relationship between O‐GlcNAcylation and p53 function in ovarian cancer. First, O‐GlcNAcylation, OGT and OGA protein and mRNA levels were measured in a series of human ovarian cancer cell lines and no correlation was found between p53 genetic status and O‐GlcNAcylation. Overexpression or silencing of p53 in a p53 wild‐type cell line (A2780) had little effect on O‐GlcNAcylation. On the other hand, pharmacological inhibition of OGA with the inhibitor Thiamet‐G (TMG) leads to an increase in nuclear localization of p53 and increased mRNA levels of p53‐target genes (p21 and Bax). Overexpression of OGA or OGT leads to increased p53 protein but not mRNA levels and a concurrent increase in p53‐targets (Mdm‐2, p21 and Bax) expression. Also, p53 acetylation on lysine 382, a transcriptional activating modification, is higher in OGA/OGT overexpressing cells. In conclusion, O‐GlcNAcylation regulates p53 function through stabilization and activation.

Support or Funding Information

Support and funding: Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Science Without Borders Program, NIH NIDDK R01 DK 100595.
BLT2 receptor protects against lethal acute lung injury induced by a pneumococcal toxin pneumolysin
Authors: Shigematsu Misako , Koga Tomoaki , Saeki Kazuko , Okuno Toshiaki , Yokomizo Takehiko
Published: November 2023

Pneumonia is one of the major causes of death over the world, and the capital cause of infectious pneumonia is S. pneumoniae (pneumococcus). The key virulence factor of pneumococcus is a pore‐forming toxin pneumolysin (PLY). By its direct cytotoxic effect, PLY is known to cause acute lung injury, leading to pneumococcal invasion and subsequent severe bacteremia. Leukotriene B4 (LTB4) receptor type 2 (BLT2) is a G‐protein coupled receptor that was originally identified as a low‐affinity receptor for LTB4, and recently we elucidated that the high affinity endogenous ligand for BLT2 is 12‐hydroxyheptadecatrienoic acid (12‐HHT), an arachidonic acid metabolite by cyclooxygenase. Previously we reported that BLT2 is expressed in intestinal epithelial cells and skin keratinocytes, and that 12‐HHT/BLT2 signaling exhibits important roles in maintaining mucosal integrity and in accelerating skin wound healing. However, the biological roles of BLT2 in other organs are still uncertified. In this study, we investigated the protective role of BLT2 in mouse lung using PLY‐induced acute lung injury model. Interestingly, BLT2‐deficient mice were more susceptible to PLY administration, and also exhibited increased vascular permeability and potent bronchoconstriction by PLY administration when compared to wild‐type mice. We found BLT2 in mouse lung is expressed in alveolar type II cells and vascular endothelial cells. The molecular mechanisms of how BLT2 exhibits the protective role for PLY‐induced acute lung injury are now under investigation.

Support or Funding Information

This work was supported by Grants‐in‐Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (MEXT) Nos. 22116001, 22116002, 15H05901, 15H05904, and 15H04708.
Bisphenol A Induces Gene Expression Related to Inflammation and Lipid Accumulation in Adipocytes
Authors: Lee Hee Weon , Kim Jinwook
Published: November 2023

Recent studies suggest that endocrine disrupting chemicals may affect obesity‐related pathway by altering cell signaling. Bisphenol A (BPA) is an endocrine‐disrupting chemical derived from polycarbonate plastics used in food and beverage containers, leading to low‐dose human exposure. BPA has the potential to affect the differentiation processes and the metabolism of the adipose tissue. In this study, we investigated the effect of BPA on adipocyte metabolic functions. 3T3‐L1 cells were incubated with BPA in the presence of MDI to examine the effects of BPA on nitrite oxide (NO) production and the expression of iNOS by colorimetric assay and Western blot, respectively. We also examined the expression levels of MAPK, NLRP‐3 and cytokines by Western blot. The results showed that BPA treatment induced NO production and the expression of iNOS. BPA also enhanced phosphorylation of ERK, JNK and p38 MAPK. Additionally, BPA treatment significantly increased the expression of NLRP‐3 and cytokines compared with the control. Moreover, we investigated the effect of adipogenesis on Lipid accumulation and the expression of adipogenic genes by Oil Red O staining and Western blot, respectively. The results showed that Lipid accumulation was increased by BPA. The expression of c/EBP β and PPAR‐γ was upregulated by BPA in adipocytes. Collectively, these data suggest that BPA acts as an enhancer of adipogenic differentiation. We suggest that BPA upregulates inflammatory signaling in adipocytes, and it is implicated in obesity and diabetes. Collectively, these data suggest that BPA acts as an enhancer of adipogenic differentiation.
Saccharomyces cerevisiae Coq10, a putative START domain protein binds CoQ and late‐stage CoQ biosynthetic intermediates
Authors: Tsui Hui Su , Clarke Catherine F
Published: November 2023

Coenzyme Q (ubiquinone or CoQ) functions as an essential redox‐active lipid in respiratory electron and proton transport in cellular energy metabolism, and it is an important lipid‐soluble antioxidant in cellular membranes. In Saccharomyces cerevisiae, at least 13 nuclear‐encoded genes COQ1‐COQ11, YAH1, and ARH1 have been identified as necessary for CoQ biosynthesis and its function in respiration and as an antioxidant. Many of the Coq polypeptides responsible for CoQ biosynthesis assemble into a high molecular weight, multi‐subunit complex localized to the matrix side of the mitochondrial inner membrane. My project focuses on characterizing Coq10, a putative steroidogenic acute regulatory (StAR)‐related lipid transfer (START) domain protein in S. cerevisiae. Unlike CoQ‐less coq1‐coq9 deletion mutants, yeast coq10 null mutants contain near wild‐type steady‐state levels of Q6, but are nonetheless respiratory deficient. Yeast coq10 null mutants show decreased de novo Q biosynthesis and accumulate high levels of the early Q intermediates, especially during early log phase of growth. The structure of CC1736, a homolog of Coq10 in Caulobacter crescentus, contains a hydrophobic tunnel characteristic of START domain proteins. Purified CC1736 is capable of binding CoQ and a farnesylated analog of a late‐stage CoQ biosynthetic intermediate. Overexpression of CC1736 in the coq10 null mutant is able to rescue the respiratory‐deficient phenotype and partially restore efficient CoQ biosynthesis, suggesting a strong functional conservation. These observations exclude Coq10 from being required for CoQ biosynthesis, but suggest Coq10 is crucial for CoQ function in respiratory electron transport and for CoQ to function as an antioxidant. Coq10 is speculated to be a CoQ‐binding protein interacting with the multi‐subunit Coq polypeptide complex and facilitating final delivery of CoQ from its site of synthesis to respiratory electron transport chain complexes. Three specific aims are proposed for characterizing the biological function of Coq10: 1) isolate and characterize the high molecular weight Coq10 oligomer; 2) characterize lipid ligand interactions with mature Coq10; 3) investigate possible phosphorylation of Coq10. The results will provide important clues about the physiological role of Coq10 as well as the molecular interactions and post‐translational modifications required for its function.
Integrating Rare Disease Research Into An Undergraduate Liberal Arts Curriculum To Enhance The Pace Of Rare Disease Gene Discovery For Underserved Populations And Promote STEM Retention
Authors: Jinks Robert N , Davis Beckley K , Roberts David M , Rice Elizabeth , Brigatti Karlla W , Puffenberger Erik G , Strauss Kevin A
Published: November 2023

Next‐generation sequencing accelerates disease gene discovery, especially for orphan diseases, though at present it outpaces functional studies needed to provide ‘proof of causation.’

Working from a small, community‐based, clinical laboratory, the Clinic for Special Children has identified more than 170 allelic variants associated with disability, disease, or untimely death among the Amish and Mennonite (Plain) populations of North America. In collaboration with the Clinic, we developed an HHMI‐funded program that integrates functional studies of novel disease gene alleles into our undergraduate curriculum. Roughly 150 students per year in our introductory cell biology and neuroscience courses clone human disease genes and study the functional impacts of gene variants through expression in mammalian cell culture. We use these authentic research experiences to teach key concepts in cell biology, genetics, and neuroscience. Students later build upon this experience in upper‐level courses in cell biology, neuroscience, genetics, cancer biology, and immunology in which they engage in semester‐long research projects in small teams. Teams conduct functional studies of disease gene variants or study novel cellular or animal models of disease. These experiences engender talented undergraduates to assume greater research responsibility through independent study and summer research projects that transition them into the role of co‐PI for their project. Data published through this project (e.g., PLoS ONE 7:e28936) have been used by other institutions to establish novel diagnoses, including a high‐profile diagnosis of a critically‐ill, non‐Plain newborn with lethal neonatal seizure‐rigidity syndrome (Sci Transl Med 4:154ra135). This provides important proof‐of‐concept for integrating novel disease gene functional studies into a carefully structured undergraduate research curriculum.

Students in our Public Health program are collaborating with the Clinic to develop handbooks to help parents care for children with special medical needs with the goal of producing one to two high‐quality disease handbooks annually.

Our program represents a model for engaging undergraduates in meaningful research at the front lines of biomedical science and public health in ways that directly impact the diagnosis and care of children with rare inherited disorders and promote STEM retention.

Support or Funding Information

HHMI Undergraduate Science Education Awards 52006294 and 52007538.
Changing career prospects are changing the needs in graduate and post‐doc training
Authors: Feeney Shea E.
Published: November 2023

The ultimate desired outcome for the acquirement of doctoral and post‐doctoral training is arguably not just the accruement of scientific knowledge but the development of a broader skill set. Providing training opportunities for those that are entering into careers as early scientists to be successful in areas beyond basic bench work is essential in crafting a productive and ethical workforce but also to accommodate the shifting and highly competitive job market. It can be considered not just detrimental but irresponsible to expect to train graduate students and post‐doctoral fellows solely for careers working at the “bench.” Furthermore, positions in academic research, once the provider of the majority of scientific careers, is at odds with the growing population of scientists granted graduate degrees. Although “alternative” career options beyond the academic research professor is now more openly discussed there is still a need to develop and incorporate skill sets into our graduate and post‐doctoral training that will enable our early scientists to be successful in a multitude of careers and professional situations. These skills transcend just accurate and reproducible performance of techniques but include critical thinking, analysis, research ethics, problem‐solving, oral and written communication, time and project management, interpersonal and conflict resolution skills—skills that are important for almost all, if not all, careers in science but not emphasized in training nearly enough. Additionally, this necessary skill set includes what preliminary results suggest a large portion of students find challenging but is perhaps one of the most important—the elusive skill of networking. Preliminary data includes survey results from graduate students and post‐doctoral fellows asked to assess the quality of training opportunities that are available at their institution and what skills they believe their training is lacking in. This is contrasted to what senior scientists in different careers consider the incoming science workforce lacks. Lastly, different approaches in broaching the gap for skill set development is evaluated to facilitate a change in training that is complementary to the changing needs that graduate and post‐doctoral scientists face.
Non‐Profit Patient Research Services from Rare Genomics Institute Enable Insight into Rare Diseases through Genome Sequencing and Cloud Collaboration
Authors: Tucker Brady , Aguilar Elizabeth , Gurnon Daniel
Published: November 2023

The non‐profit Rare Genomics Institute (RG) helps provide rare disease patients with access to cutting‐edge molecular biotechnology and analytical expertise with the potential to uncover new directions for research, treatment, and support. As volunteer analysts for RG, we analyzed the exome sequence data of patients with as‐yet undiagnosed diseases with a suspected underlying genetic cause. Data analysis is performed with Omicia Opal, a web‐based genome interpretation and reporting software platform integrating the machine learning algorithms VAAST and Phevor. We summarize our use of the Omicia Opal platform in three cases that represent the range of outcomes that can result from exome analysis; in one case we identified a likely pathogenic variant in the RDH12, associated with Leber's congenital amourosis; in a second case we identified a possibly pathogenic variant in IFT140, associated with Jeune syndrome; and in a third case we identified variants of uncertain significance in genes associated with Sotos and Weaver syndrome. We also outline the additional work RG analysts do to prepare an informative report for the family and their physician, and to connect the family and clinicians to experts who may be able to move their case forward.

While definitive answers are still uncommon in exome analysis, the research of RG analysts can provide new leads in a difficult case. Moreover, undergraduate students can make meaningful contributions to patient‐centered rare disease research provided that expert guidance and oversight are in place.

Support or Funding Information

Funding provided by DePauw University
Molecular Visualization in the Classroom: Learning Goals and Competencies
Authors: Dries Daniel R. , Craig Paul A. , Dean Diane M. , Listenberger Laura , Novak Walter R.P. , Franzen Margaret A
Published: November 2023

Biochemical educators agree that visual literacy is an essential tool for learning and as such should be explicitly taught and assessed. However, few resources exist for educators to examine and improve teaching or assessing visual literacy. We have utilized our collective expertise in teaching biomolecular science courses to expand the framework for 3D literacy initially presented by Craig and Bateman. We identify visual literacy skills to be mastered by novices (first year life science students), amateurs (upper division undergraduates) and experts (graduate students or professionals in the field). Moreover, we have begun work to unpack these skills with the goal of facilitating the development of both formative and summative assessments. We encourage the BMB community to use the document presented here to facilitate the inclusion of visualization skills in curriculum development in line with Vision & Change and the ASBMB recommendations. Furthermore, we seek to identify other scientists and educators with an interest in participating in online resource development of 3D literacy materials.

Support or Funding Information

This material is based upon work supported by the National Science Foundation under award number DUE‐1323414. Any opinions, findings, and conclusions or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the views of the National Science Foundation.
Overweight and obesity among women in a predominantly rural district of Ghana
Authors: Marquis Grace S , Colecraft Esi K , Bannerman Boat , Thompson‐Colón Theresa
Published: November 2023

The double burden of malnutrition is evident among women of reproductive age (WRA) in Ghana where micronutrient deficiencies commonly occur concurrent with an increasing prevalence of overweight and obesity (OW/OB). However, overnutrition among Ghanaian women has been rarely characterized. We assessed sociodemographic determinants of OW/OB among women in a predominantly rural district of Ghana. Cross‐sectional data were collected from WRA (n=1095) who lived in the Upper Manya Krobo District of the Eastern Region and who participated in the Nutrition Links Infant Household baseline survey between November 2013 and May 2014. The women were interviewed about their personal and household characteristics and weight and height measurements were taken. Body mass index (BMI) was calculated and a dichotomous variable was used to represent non‐OW/OB (BMI<25 kg/m2) and OW/OB (BMI ≥ 25 kg/m2). The mean age of the women was 27.4 ± 7.6 y. Over one‐quarter of them were either overweight (21%) or obese (6.4%); the prevalence of underweight (7.2%) was similar to that of obesity. OW/OB was significantly associated with being 30 y or older, married, and having more household assets (at least 4 out of 12 items) (p<0.05). However, those women who owned their home were less likely to be OW/OB (p<0.05). Combined, the prevalence of overweight and obesity was almost four‐fold higher than that of underweight among these predominantly rural women. Concurrent efforts to address both over‐ and undernutrition are needed to enhance women's health and well‐being in this population.

Support or Funding Information

Funded by the Department of Foreign Affairs, Trade, and Development (DFATD) of the Government of Canada
Scant interactions of biomarkers of oxidative stress and anti‐oxidation defense with those of the inflammatory response among preschool children form the Western Highlands of Guatemala
Authors: Méndez Maria José Soto , Aguilera Concepción María , Mesa Maria Dolores , Rico María Cruz , Campaña‐Martín Laura , Martín‐Laguna Victoria , Solomons Noel W. , Schuemann Klaus , Gil Angel
Published: November 2023

Objective

To explore the presence and magnitude of paired associations between diagnostic biomarkers of oxidation and inflammation from the same or different anatomical compartments.

Methods

82 children, 38 girls and 44 boys (median age 56 mo), attending 3 daycare centers from a government‐subsidized system in Quetzaltenango, Guatemala, delivered single samples of urine, whole blood, saliva, and feces for one or more of the selected determinations. We were able to make pairing correlations of Among 23 biomarker variables, paired Spearman rank‐order correlations could be made from 12 from the inflammatory domain (White blood cells, plasmatic and salivary IL‐1β, IL‐6, IL‐8, IL‐10 and TNFα, and fecal calprotectin) and 11 from the oxidation domain (urinary markers of oxidation {15‐Isoprostane‐F2t and 8‐hydroxy‐deoxy‐guanosine}; erythrocyte activity of antioxidant enzymes {catalase, superoxide dismutase, glutathione reductase and glutathione peroxidase}; and plasmatic concentrations of antioxidant nutrients {retinol, tocopherols, β‐carotene and Coenzymes Co‐Q9 and Co‐Q10}). The SPSS version 20 software program was used.

Results

Within the 132‐cell full matrix, 13 statistically‐significant associations were found (9.8% of possible), seven at a p‐value of <0.01 and 6 from <0.05 to 0.01. Superoxide dismutase and β‐carotene were each associated with 5 and 6 inflammatory biomarkers, respectively. Backward‐elimination multiple‐regression analyses with both as dependent variables showed the same two predictors, plasmatic TNF‐α and saliary IL‐8, with an r2‐value of 0.155 for superoxide dismutase and r2 0.136 for β‐carotene.

Conclusion

Significant associations in an inter‐class context between paired, oxidation‐vs‐inflammation biomarker variables represent only a 10th of the possible associations and a fraction of the 33–57% association seen with the same data, previously analyzed on an intra‐class hemi‐matrix basis.

Support or Funding Information

Funded by: Fundación Iberoamericana de Nutrición (FINUT), Spain and The Hildegard Grunow Foundation (HGF), Germany
The role of dairy in the comparative effectiveness and cost of fortified blended foods versus ready‐to‐use foods in treatment of children with moderate acute malnutrition
Authors: Suri Devika J , Moorthy Denish , Rosenberg Irwin H.
Published: November 2023

Objective

Recent meta‐analyses found treating young children with MAM using ready‐to‐use foods (RUF) versus fortified blended foods (FBF) resulted in higher recovery rates and weight gain. This analysis aimed to compare studies of RUF and FBF with and without dairy to determine whether the addition of dairy to these food supplements modified the comparative effectiveness and cost of treatment.

Methods

A review of literature on the comparative effectiveness of FBF and RUF in treatment of MAM was conducted. Outcomes of recovery from MAM, weight gain and length gain were compared among study cohorts, which included FBF with dairy (FBF+), FBF without dairy (FBF−), RUF with dairy (RUF+) and RUF without dairy (RUF−). Data on recovery from MAM was pooled among the 4 supplement categories. The cost per 500 kcal of each category of food supplement was averaged among studies that reported cost data.

Results

Among the 7 studies included, 9 RUFs were tested, of which 5 contained dairy, and 9 FBFs were tested, of which 3 contained dairy. Children treated with RUF+ had higher recovery rates compared with FBF− in 5 out of 5 study cohorts, higher weight gain in 4 out of 4, and significantly higher length gain in 1 out of 4. Children treated with RUF+ vs FBF+ had higher recovery rates in 1 out of 2 study cohorts, with no differences in weight or length gain. No differences were found in the 2 studies comparing RUF− and FBF+. Finally, children treated with RUF− had higher recovery rates compared with FBF− in 1 of 2 studies, higher weight gain in 2 out of 2, and no differences in length gain. Recovery from MAM among the 7 studies was 65% (FBF−), 79% (FBF+), 82% (RUF−), and 80% (RUF+). Four of the 7 studies included cost data; on average per 500 kcal costs were $0.15 (FBF−), $0.18 (FBF+), $0.17 (RUF−), and $0.35 (RUF+).

Conclusion

Our results suggest that addition of dairy to FBF make it comparative in effectiveness to both RUF with and without dairy, but does not appear to be a factor between the RUF categories. RUF with dairy was twice the cost per kcal compared with the other food supplement categories. Cost‐effectiveness analysis will be useful to help determine the most appropriate food supplement for treatment of MAM.

Support or Funding Information

This work was supported in part through a grant from the Food For Peace office of USAID
Contribution of School Meals to Nutrient Intakes of School‐age children enrolled in two feeding programs in Ghana
Authors: Owusu Justina Serwaah , Colecraft Esi , Aryeetey Richmond , Vaccaro Joan A. , Huffman Fatma
Published: November 2023

School‐age children are vulnerable to malnutrition which can negatively influence their growth and maturity, health and academic attainment. School feeding programs offer an opportunity to reduce hunger and malnutrition among school age children. However, there is limited information on how school meals contribute to dietary intakes of school children in Ghana. We compared the contribution of meals provided by ‘Ghana School Feeding Programme’ (GSFP) and a ‘Non‐governmental School Feeding Programme’ (NSFP) to total energy and nutrient intake of school‐age children. Two non‐consecutive 24‐hour dietary recalls were completed with 180 school age children. Additionally, the weighed food method was used to obtain weights of 7 dished portions of school meals served daily over a one‐week period at both feeding sites. Program differences were assessed using independent t‐test. The school‐age children in the ‘Ghana School Feeding Programme’ received significantly smaller portions of the school meal than those in the ‘Non‐Governmental School Feeding Programme’ (243 ± 50g vs. 416 ± 96g; p=0.007). Whereas the ‘Ghana School Feeding Programme’ meals consumed by school‐age children contributed from 12% up to 18.1% of the total energy and nutrient intakes of the children, the ‘Non‐governmental School Feeding Programme’ meals contributed from 17% up to 30% of the total energy and nutrient intakes of the children. The ‘Non‐governmental School Feeding Programme’ contributed significantly more energy (28 ± 10% vs. 16.2 ± 7%; p=<0.001), protein (24.6 ± 9% vs. 13.3 ± 7%; p=<0.001) and micronutrients (p≤0.042) to the children's total intakes compared to meals provided through the ‘Ghana School Feeding Programme’. The smaller portion sizes provided by the GSFP contributed to the meals supplying less energy and nutrients to children's daily intakes. The quality of meals served by both programs needs to be reviewed to ensure that they supply the recommended 30% of energy and nutrients to school children's daily requirements.

Support or Funding Information

The project was funded in part by Prembaf Ghana, an affiliate of Prem Rawat Foundation.