bluuebunny/crossref_metadata_2025_split huggingface dataset:
Annotating the CIA7 Gene: Comparing Lead Bioaccumulation, Cell Growth and Morphology in Two Strains of Chlamydomonas reinhardtii
Authors: Gutierrez Jose Angel , Ynalvez Ruby 
Published: November 2023

As industrialization increases to meet the demands of 21st century life, incidences of heavy metal contamination increases as well. Non‐essential heavy metals serve no biological function and can have devastating effects across all spectra of life. Lead (Pb) is of significant research importance due to its toxicity at both acute and chronic levels of exposure. Industrial sources of lead contamination include production of crude oils, and paints. Bioavailable Pb contamination can damage key metabolic processes in eukaryotic and prokaryotic organisms. Plants and algal cells are particularly affected by Pb, as unchecked exposure can cause metabolic dysfunction and eventually damage the chloroplast's photosystems. Alga and plants employ an array of physical and chemical defenses to maintain heavy metal homeostasis. Among these are the production of non‐specific heavy metal binding proteins known as phytochelatins (PC) and metallothioneins (MT). Both protein families share similar amino acid motifs, particularly moieties of cysteine, whose sulfhydryl (‐SH) active site has a high affinity for heavy metal ions. MT and PC aid in the chelation and eventual removal of heavy metal ions. Investigation of heavy metal tolerance genes in green algae is of importance due to their position as primary producers in most aquatic ecosystems. Chlamydomonas reinhardtii, a unicellular green alga, has been a model organism for the study of photosynthesis, bioremediation, and heavy metal toxicity. Though its genome has been sequenced, several genes and their protein products remain unannotated. One such gene, Cia7, is hypothesized to play a role in maintaining C. reinhardtii's heavy metal homeostasis. Preliminary evidence shows that CIA7 contains a highly conserved motif similar to other metal sequestering proteins. To aid in confirming CIA7's ability to sequester heavy metals, this study presents the results of a bioaccumulation experiment involving a wild type (CC4425) and a Cia7 mutant (CC5013). CC4425 and the CC5013 were grown simultaneously in separate treatment flasks containing either 0uM/150 uM PbCl2 in replete TAP media for 4 days under constant lighting. Pb ion bioaccumulation was measured utilizing an Induced Coupled Plasma Optical Emission Spectrophotometer (ICP‐OES) following a microwave acid digestion step. A 2×2 factorial analysis of variance was conducted to determine the significance of the data obtained. Results of bioaccumulation experiments indicated that there is a significant difference in Pb bioaccumulation between the mutant cia7 and wild‐type at 150 μM Pb treatment. The mutant cia7 bioaccumulated higher concentration of Pb compared to wild‐type. This difference is significant at 95% confidence interval. Flow cytometry and fluorescent microscopy experiments were conducted to further explore the differences in morphology and cell count between strains across both treatments.Support or Funding InformationNorman Hackerman Advanced Research Program
Glucocorticoid‐induced inhibition of AKT leads to CREB phosphorylation and increased myostatin expression via a PDE/cAMP/PKA pathway in skeletal muscle
Authors: Xie Yang , Zhang Peng , Espinoza Daniel , Perry Ben , Rahnert Jill , Zheng Bin , Price Russ 
Published: November 2023

Muscle atrophy in diseases like chronic kidney disease and diabetes is associated with dysfunctional regulation of insulin/Akt/FoxO3 and myostatin‐SMAD3 signaling pathways. We recently found CREB, a transcription factor that regulates maintenance and regeneration of skeletal muscle, is phosphorylated in wasting conditions. In silico analysis identifies a CREB binding site in the myostatin promoter. This study investigated the mechanism by which atrophy‐inducing conditions regulate CREB phosphorylation and its impact on myostatin expression. Differentiated C2C12 myotubes were incubated with combinations of dexamethasone (Dex), LY294002 (PI‐3 kinase inhibitor), H89 (protein kinase A inhibitor), isobutylmethyl xanthine (IBMX, a general phosphodiesterase (PDE) inhibitor), milrinone (PDE3 inhibitor), or rolipram (PDE4 inhibitor) for up to 24 h. Cells were harvested for analysis of proteins or mRNAs. Treatment of myotubes with Dex for 6–24h increased CREB phosphorylation (pCREB) within 3 h and sustained the increase over 24h. Dex also inhibited Akt phosphorylation (pAkt) over a similar time frame. Inhibition of pAkt by LY294002 mimicked Dex by increasing pCREB; the actions of LY294002 and Dex on pCREB were not additive. Since Akt can activate PDE3/PDE4 and reduce cAMP/PKA signaling in non‐muscle cells, we tested whether Dex‐induced inhibition of PDE3/4 increases pCREB. Without Dex, IBMX, milrinone, and rolipram increased pCREB. The Dex‐induced increase in pCREB was attenuated by the PKA inhibitor H89. When added individually, Dex, milrinone, rolipram and IBMX increase myostatin mRNA and protein significantly. Our study identifies a new mechanism by which atrophy‐inducing conditions activate CREB and contribute to myostatin expression. Signals that attenuate insulin/Akt signaling reduce the PDE3/4 activity. This leads to an increase in cAMP, PKA activity, CREB phosphorylation and myostatin expression.Support or Funding InformationFunded by NIH RO1 DK95610 and VA Merit X01BX001456
Puringeric Signaling in Bone as a Potential Mechanism in Prostate Cancer Proliferation and Cancer‐Induced Bone Pain
Authors: Wilson Michael , Duncan Randall , Boggs Mary 
Published: November 2023

Prostate cancer (PCa) is the second leading cause of cancer‐related deaths in men in the US. PCa bone metastasis occurs in approximately 2 out of 3 patients whose cancer has spread to other parts of the body. In particular the majority of PCa that metastasizes to bone results in osteosclerotic tumors. Osteosclerotic tumors arise from interactions between cancer cells and osteoblasts that results in increased bone formation, but reduced bone strength and increased mechanical strains. The increased load in bone resulting from tumor formation correlates with increased pain sensation. Cancer‐induced bone pain (CIBP) is one of the most detrimental symptoms to patients with bone‐localized cancer, greatly reducing patient's quality of life. Current treatment for CIBP is ineffective. A better understanding of the molecular mechanisms of CIBP is necessary for proper treatment. We propose that mechanical load of PCa cells results in ATP release which can activate local neurons through P2X3 receptors resulting in CIBP. Additionally, we propose that nerve‐growth factor (NGF), a molecule implicated in PCa proliferation, enhances the potential for CIBP by through PCa cell proliferation and ATP signaling. To determine the effect of mechanical load on ATP release from PCa cells and activation of neurons, we mechanically loaded human bone‐derived PCa cells, C4‐2B4, via hypotonic swelling. Resultant C4‐2B4 load‐response conditioned media (L‐CM) +/− apyrase or various concentration of ATP were applied to primary mouse dorsal root ganglia (DRG) and changes in intracellular calcium were measured. To determine the role of P2X3 in response to L‐CM, TNP‐ATP, a P2X3 specific antagonist was utilized. Data indicated that mechanical load increased ATP release from C4‐2B4 cells and that L‐CM increased intracellular calcium in DRG which was reduced with apyrase. TNP‐ATP did not significantly block response to L‐CM suggesting another purinergic receptor may be responsible for this response.To address the role of NGF in PCa proliferation and purinergic signaling, we applied NGF to C4‐2B4 and counted cells daily for 4 days. To determine the effect of NGF on C4‐2B4 response to ATP, we applied various concentrations of ATP to C4‐2B4 cell pretreated with NGF for 48 hrs and measured intracellular calcium. NGF treatment demonstrated a 32% increase in cell number at day 4 when compared to control. Calcium imaging studies indicated that response to ATP varied with addition of NGF indicating a potential pathway for altered sensitivity to ATP. These studies indicate that NGF may play an important role in the growth of metastatic prostate cancer cells in bone. Taken together these data indicate purinergic receptors as a potential therapeutic target for CIBP.Support or Funding InformationDelaware INBRE programNIH‐NIGMS (8 P20 GM103446‐15)Charles Peter White Fellowship
Elucidating the Reproductome: System‐wide Regulation of Reproductive Neuropeptides
Authors: Glucksman Marc , Philibert Keith , Woitowich Nicole , Urban Janice , DeJoseph Gina 
Published: November 2023

Kisspeptin (Kiss), neurokinin B (NKB) and dynorphin A (DynA), collectively known as the KNDy neurons in the hypothalamus, drive the activity of the reproductive axis through the regulation of gonadotropin releasing hormone (GnRH) release. These neurons project to the rostral hypothalamus and median eminence to relay peripheral signals such as estrogen levels and energy homeostasis to the GnRH neuron. As neuropeptides, the actions of Kiss and DynA are required to be modified enzymatically in the extracellular space to modulate downstream signaling, though the regulator of this process is unknown. Furthermore, Phoenixin, a recently identified neuropeptide produced in the hypothalamus, increases GnRH, GnRH receptor and Kisspeptin mRNAs, increases GnRH secretion and mediates ovarian cyclicity. EP24.15 (EC 3.4.24.15, thimet oligopeptidase) is a metallo‐endopeptidase, expressed throughout the hypothalamus and other brain regions, that cleaves GnRH and inactivates binding to its cognate receptor.To authenticate a biochemical and physiological regulatory mechanism for the functioning of Kiss and DynA, we utilized a high throughput workflow schema by first identifying potential active site binding and cleavage sites both in silico and by mass spectrometry; measured enzymatic parameters; performed structural studies. Finally, we determined if EP24.15 and both Kiss and DynA are co‐expressed in the brain, leading to the elucidation of a functional neuroendocrine circuit whereby EP24.15 could regulate Kiss‐ or DynA ‐induced reproductive hormone secretion. EP24.15 and Kiss were co‐incubated and subjected to MALDI‐TOF mass spectrometry to confirm that EP24.15 cleaves full length Kiss as well as the fully functional Kiss10 decapeptide between S5–F6 in vitro (Vmax = 2.37±0.09 pmol/min/ng, Km = 19.68±2.53 μM). Dynorphin A (1–8) is cleaved between F4–L5 and L5–R6 (Vmax = 30.9±0.48 pmol/min/ng, Km = 14.62±0.89 μM). Phoenixin is cleaved between K8–V9 in vitro (Vmax = 2.90±0.20 pmol/min/ng, Km = 25.81±5.40 μM). These novel substrates are cleaved at rates comparable to other known neuropeptides. We have also demonstrated that both Kiss and DynA immunoreactivities are co‐expressed and localized with EP24.15 in the hypothalamus. These data demonstrate that EP24.15 cleaves Kiss, DynA, and Phoenixin in vivo to unveil an additional layer on the exquisite control mechanism of the mammalian reproductome. Further understanding the role of EP24.15 in neuropeptide metabolism will provide insight into the circuitry within the hypothalamus and provide targets for modulating various neuroendocrine axes, including reproductive function.Support or Funding InformationSupported by NIH OD010662 (MJG).
Antimicrobial activity of a high‐pressure enzymatic extracts using Prunus mume
Authors: Jang Jong‐Hwa , Oh Tae‐Jin 
Published: November 2023

The most common oral diseases are dental cavities and periodontal diseases, which are infectious diseases. Antimicrobial agents for the prevention and treatment of oral disease as alternatives to conventional oral antimicrobial agents, such as chlorohexidine, have been investigated. Numerous natural compounds have been reported to be effective in preventing and treating oral infections, and in maintaining good oral health. Among them, Prunus mume, known as Japanese apricot, is used as a traditional medicinal food in Korea, China, and Japan. However, the common methods of Prunus mume extraction, including solvent or aqueous thermal‐based extraction, result in low yields, long extraction times, and traces of organic solvents. Enzymatic extraction of bioactive compounds from plants is a potential alternative to conventional solvent or aqueous thermal‐based extraction. Enzymatic treatment is used in the food industry due to its low operating temperature and ability to increase extraction yields. High‐pressure processing is a novel technology that has shown potential for extraction of bioactive components. Therefore, the bioactive components are more easily washed out, such as flavonoids, alkaloids, vitamins, saponins and pigments. The high‐pressure extraction technique is associated with a short extraction time, high extraction yield and lower levels of impurities. Our goal was to evaluate high‐pressure processing combined with enzymatic treatment for extraction of Prunus mume by determining the optimum extraction conditions for the antimicrobial activity against oral bacteria. The maximum antimicrobial activity of Prunus mume extract obtained using Novozym 33095 was at pH 7.0, 45°C, 20 h, and 5 v/v%. Minimum inhibitory concentration values were 3.125 to 12.50%. These results indicated that high‐pressure enzymatic extraction yielded Prunus mume extract with antimicrobial activity which has the potential for improving oral environment.Support or Funding InformationAcknowledgement:This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (2014R1A1A3051084) and was supported by Business for Cooperative R&D between Industry, Academy, and Research Institute funded Korea Small and Medium Business Administration in 2014 (C0190937). In addition, this work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ01045705)” Rural Development Administration, Republic of Korea.
Type 2 Diabetes‐Associated Variants Disrupt Function of SLC16A11, a Proton‐Coupled Monocarboxylate Transporter, Through Two Distinct Mechanisms
Authors: Hoch Eitan , Rusu Victor , Schreiber Stuart L. , Florez Jose C. , Jacobs Suzanne B. R. , Lander Eric S. 
Published: November 2023

IntroductionType 2 Diabetes (T2D) affects more than 415 million people and is a leading cause of morbidity and mortality worldwide. While T2D is influenced by environmental factors, it is also a highly heritable disorder, with genetic variation contributing to a disparity in T2D prevalence across populations. An example of this disparity is observed within American populations, where the prevalence of diabetes in individuals of Mexican or Latin American descent is approximately twice that of US non‐Hispanic whites. Through a genome‐wide association study, we recently identified a variant haplotype in SLC16A11 that explains ~20% of the increased T2D prevalence in Mexico.ObjectiveTo follow up on the genetic association at SLC16A11, in order to delineate mechanisms underlying T2D risk at this locus.ResultsUsing a combination of molecular, biochemical, cellular, and physiological approaches we identify two distinct mechanisms that lead to reduced SLC16A11 function. First, we observe a significant reduction in SLC16A11 expression ‐ in a dose‐dependent manner ‐ in carriers of the T2D risk haplotype, in human liver. Second, we demonstrate that T2D risk‐associated coding variants in SLC16A11 attenuate activity by disrupting a key interaction with an ancillary protein, thereby reducing plasma membrane localization. These two independent mechanisms by which T2D‐associated coding and non‐coding variants impact SLC16A11 expression levels and subcellular localization implicate perturbation of SLC16A11 as causal at this locus, and suggest reduced SLC16A11 activity as the T2D‐relevant direction‐of‐effect.To gain insight into how disruption of SLC16A11 function impacts T2D risk, we investigate the activity of this previously uncharacterized transporter and establish that SLC16A11 functions as an H+‐coupled monocarboxylate transporter.ConclusionOur findings illustrate the path from genetic association to therapeutic hypothesis, through defining the molecular mechanisms by which genetic variation affects SLC16A11 action, and suggest that increasing SLC16A11 function could be therapeutically beneficial for people with T2D.Support or Funding InformationThis work was conducted as part of the Slim Initiative for Genomic Medicine, a project funded by the Carlos Slim Foundation in Mexico
Right Time to Sprinkle Pepper
Authors: Lee Jee Yeon , Choi A‐ri , Kim Dohee , Yon Miyong , Nam Jiwoon , Park Seungjoo , Lee Haeng‐Shin , Lee Gae Ho , Lee Kyung Jun , Kim Cho‐il 
Published: November 2023

Acrylamide (AA) in foods was first identified by the Swedish National Food Authority in 2002. Since then, WHO, EU, US, and others conducted AA monitoring on foodstuff and found high levels in potato chips, french‐fries, coffee, etc. Although AA is produced unavoidably by heating foods of high carbohydrate and low protein content at high temperature, it deserves special public health attention because the International Agency for Research on Cancer of WHO has classified AA as carcinogenic to humans (Group 1). Therefore, we conducted total diet study (TDS) to estimate AA exposure of the Korean population using the dietary intake data collected by 24‐hr recall method from Korea National Health and Nutrition Examination Survey (KNHANES). Data from 2008 through 2013 KNHANES were merged to increase the statistical power while giving equal weights to each year data and analysed to extract the representative food (RF) list for AA analysis. Foods consumed most by Koreans were lined up by intake amount and frequency and a list of 543 foods covering 97.4 % of mean intake was constructed. Based on the foods/dishes for which these RFs were used as ingredients, cooking method for each ingredient food was coupled to make ‘food □ cooking method’ samples. A total of 1224 ‘food □ cooking method’ samples were prepared using foods collected by nationwide sampling from 9 metropolitan cities for AA analysis. AA was analysed by LC/MS after solid phase extraction. AA was detected in 8.8% of analysed samples and black pepper samples showed the highest content. Top 5 on the list were all black pepper samples; ‐baked (12,029.1 ppb), ‐fried (3,595.6 ppb), ‐stir‐fried (1,313.2 ppb), ‐boiled (1,169.4 ppb), and ‐pan‐fried (1,083.7 ppb). Granulated instant coffee (507.0 ppb), french‐fries (463.7ppb), and potato chips (344.8~394.1 ppb) followed while black pepper‐ as is (324.1 ppb) showed a relatively low content. That is, heating in different cooking methods resulted in a 3 to 37 fold increase in AA production. This may explain why previous attempts to correlate AA exposure with the incidence of cancer were not quite successful. On the other hand, this information is valuable to the food industry and to individual consumers seeking to reduce AA levels in food. Mean AA exposure of the Korean population was estimated to be 0.09 μg/kg BW/day corresponding to 1/6 –1/11 of the values reported in US or Australia. The right time to sprinkle black pepper is 『just before consumption, at the table!』.Support or Funding InformationThis research was supported by a grant (16162MFDS116) from Ministry of Food and Drug Safety in 2013–2016.
Breastfeeding counseling improves maternal‐infant feeding interaction in those exposed to controlling behavior or emotional violence: MINIMat study in Bangladesh
Authors: Frith Amy , Ziaei Shirin , Frongillo Edward , Khan Ashraful , Ekstrom Eva‐Charlotte , Naved Ruchira 
Published: November 2023

Good quality maternal‐infant interaction is foundational for infant health and development, but domestic violence may impede on women's ability to interact well during feeding. Further, while breastfeeding counseling benefits breastfeeding duration, its influence on maternal‐infant feeding interaction is unknown in vulnerable populations.We aimed to determine if the influence of breastfeeding counseling on maternal‐infant feeding interaction depended on maternal exposure to controlling behavior or emotional violence during pregnancy. We used data from 180 mother‐infant dyads (infants 3–4 m old) who were part of a sub‐study of the Maternal and Infant Nutrition Intervention (MINIMat) study in Matlab, Bangladesh. Women were randomly assigned at 30 weeks of gestation to 1 of 2 counseling groups: usual health message counseling (UHM) or exclusive breastfeeding counseling with usual health message counseling (BFC). Lifetime exposure to controlling behavior and pregnancy exposure to emotional violence were measured by the modified version of Conflict Tactic Scale at 30 wk gestation. NCAST scales (total, maternal or infant total, and maternal sensitivity and social‐emotional fostering scales) were used to measure maternal‐infant feeding interaction.There were interactions of counseling and exposure to controlling behavior on total NCAST (P‐interaction < 0.08) and maternal total (P‐interaction < 0.05), sensitivity (P‐interaction < 0.04), and social‐emotional fostering (P‐interaction < 0.02) scales. After controlling for food security, there were interactions of counseling group and exposure to emotional violence during pregnancy on total NCAST (P‐interaction <0.05), maternal total (P‐interaction <0.04), maternal sensitivity (P‐interaction <0.1), and maternal socio‐emotional fostering (P‐interaction <0.04). For those who had experienced controlling behavior or emotional violence during pregnancy, BFC improved maternal‐infant feeding interaction to a greater extent than in those who did not experience these forms of violence, or those who had but were in the UHM group.Breastfeeding counseling is beneficial to maternal feeding behaviors that promote infant development for those who are vulnerable because they have experienced domestic violence.Support or Funding InformationSource of funding: The MINIMat research study was funded by ICDDR,B, United Nations Children's Fund (UNICEF), Swedish International Development Cooperation Agency (Sida), UK Medical Research Council, Swedish Research Council, Department for International Development (DFID), Japan Society for the Promotion of Science, Child Health and Nutrition Research Initiative (CHNRI), Uppsala University and United States Agency for International Development (USAID).