bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of fermented rooibos on sperm from infertile men in vitro
Authors: Opuwari Chinyerum Sylvia , Takalani Ndivhuho Beauty , Adefolaju Gbenga Anthony , Henkel Ralf 
Published: April 2020

Rooibos (Aspalathus linearis) is indigenous to the Cederberg region in the Western Cape Province of South Africa. It is produced as unfermented and fermented rooibos tea. Previous studies demonstrated improved sperm parameters in male rats owing to its antioxidant properties. This study aims to investigate the effects of fermented rooibos in proven infertile men in vitro.Semen samples were collected after 3–5 days of abstinence from 50 patients attending the infertility clinics of Tygerberg and Vincent Palotti Hospitals, South Africa. After liquefaction, semen was washed with HTF‐BSA (1:5), centrifuged for 10 minutes at 500 ×g and exposed to aqueous extract of fermented rooibos (0, 0.10, 1, 10 and 100 μg/ml) for 1h at 37°C. Thereafter, sperm parameters such as motility, vitality, mitochondrial membrane potential, ROS production, capacitation and acrosome reaction were analyzed.Our results showed no effect of the aqueous extracts at any concentration used on the respective sperm parameters. In conclusion, fermented rooibos has no direct effect on the human sperm in vitro. Hence, the positive effect observed in previous studies may rather be due to the systemic antioxidant effect on the male reproductive system in vivo.Support or Funding InformationThuthuka fundinding instrument, National Research Foundation, South Africa (TTK170426228930)
The Role of the Fibrotic Scar in Repair Following Neuroinflammation
Authors: Dorrier Cayce , Aran Dvir , Haenelt Ezekiel , Lizama Carlos , Cautivo Kelly , Sheehy Ryan , Weiner Geoffrey , Arnold Thomas , Daneman Richard 
Published: April 2020

Multiple sclerosis (MS) is a neuroinflammatory disease of the central nervous system (CNS) in which the body’s immune system attacks the myelin sheath that surrounds and insulates axons. In many cases this myelin is not repaired by myelinating oligodendrocytes, which decreases the efficiency of action potential conduction and leads to neural dysfunction. We hypothesized that a barrier preventing oligodendrocyte lineage cells from repairing damaged myelin is a fibrotic scar. Following CNS injury, a scar consisting of an outer glial scar made up of reactive astrocytes and an inner fibrotic scar made of proteins such as collagen I forms around the site of trauma. In MS the glial scar has also been characterized, but the presence of a fibrotic scar has not been investigated. I have shown that following induction of experimental autoimmune encephalomyelitis (EAE) in mice, which is used as a model of MS, an extensive fibrotic scar forms in the lesioned tissue. Scar‐forming cells were visualized in this tissue using a Col1a1GFP mouse model. The number of these cells increased rapidly in the lesion site following symptom onset and remained high throughout the course of the disease. Lineage tracing and single cell RNA sequencing determined that these cells arise from the proliferation of fibroblasts and not other cells such as pericytes turning on the production of collagen. The objective of this project is to determine the role of the fibrotic scar in tissue repair and to determine mechanisms of scar formation in the CNS. To determine the role of the fibrotic scar following EAE I used the herpes thymidine kinase system to ablate proliferating fibroblasts. Using this paradigm I was able to reduce scar formation by 60% in EAE and found that this reduction resulted in a decrease in motor symptoms in the later stages of disease concurrent with an increase in oligodendrocyte lineage cells in the inflammatory lesions. To understand the signaling pathways that play a role in CNS scar formation, I used FACS to purify Col1a1GFP+ fibroblasts from spinal cords of healthy mice and mice with EAE and analyzed their transcriptome by RNA sequencing. I found that these cells upregulate inflammatory signaling pathways in disease such as the interferon gamma pathway. Deleting the interferon gamma receptor in CNS fibroblasts resulted in a decrease in scar formation following EAE and may be a potential therapeutic target for CNS disorders with fibrotic scarring. In conclusion I identified a fibrotic scar forms following neuroinflammation that arises from the proliferation of CNS fibroblasts, plays a role in disease recovery and forms in part through interferon gamma signaling.Support or Funding InformationThis work was funded by a NIH T32 Pharmacological Sciences Training Grant
Fold‐Switching Sets the Stage for Cooperativity and Competition in the Cyanobacterial Circadian Clock
Authors: Partch Carrie , Swan Jeffrey , Heisler Joel , LiWang Andy 
Published: April 2020

Circadian rhythms are an ancient evolutionary adaptation found across the domains of life that synchronize behavior and physiology to the daily solar cycle for optimal fitness. In cyanobacteria, timekeeping is generated by three Kai proteins (KaiA, B and C) that form distinct protein complexes throughout the day, comprising a molecular clock that measures time in ~24‐hour increments. We recently showed that KaiB undergoes a rare transition from a tetrameric ground state to a monomeric, fold‐switched signaling state that binds to KaiC and plays a central organizing role in the evening phase of the clock. Using a version of KaiB locked into its active, fold‐switched state, we determined the structures of several complexes that illustrate how KaiB docks on the KaiC hexamer to recruit KaiA and the output protein CikA. A new structure of the output protein SasA bound to KaiC suggested that structural mimicry of the KaiB fold‐switched state is utilized to transition through distinct phases of the clock. Biochemical studies demonstrate that structural mimicry is used at low concentrations of SasA to cooperatively recruit KaiB and enhance oscillator function, while it leads to competition for KaiC at high concentrations. Collectively, this work reveals how mutually exclusive binding interfaces are used to regulate day/night transitions as well as both input and output signaling in the cyanobacterial circadian clock.Support or Funding InformationNational Institutes of Health R01 grants GM121507 (to C.L.P.) and GM107521 (to A.L.).
The biological activity of extracellular matrix mimetic elastin‐like polypeptide‐laminin α1 peptide
Authors: Truong Anh Tan , Hamada Keisuke , Yamada Yuji , Guo Hao , Kikkawa Yamato , Okamoto Curtis Toshio , MacKay J. Andrew , Nomizu Motoyoshi 
Published: April 2020

The extracellular matrix (ECM) is comprised of a large network of proteins essential for providing a physical scaffold as well as biochemical and biomechanical signals for tissue development and repair. Laminins are a major component of the ECM and have a diverse set of biological roles such as cell adhesion, migration, proliferation and differentiation. Laminin α1 chain bioactive RGD containing peptide, A99 (AGTFALRGDNPQG), which was previously identified to promote strong cell attachment has demonstrated utility for cell adhesion applications. A wide array of biomaterials has been created to be used as scaffolds for these types of bioactive peptides, but many are not suited for various reasons such as scalability, convenient reaction conditions or lack of homogeneity. Here, we are the first to describe methods for preparing chemically conjugated and recombinant A99 to elastin‐like polypeptides (ELPs) as the scaffold and characterize their behavior as well as evaluate the coating efficiency, cell attachment and spreading activity. These data suggest that the A99‐ELP fusion has great potential for use as a biomaterial in cell culture applications as they greatly promote cell adhesion and spreading. ELPs are biocompatible protein‐polymers that are thermo‐responsive and self‐assemble in response to heat. Below the lower critical solution temperature (LCST), they are highly soluble but above the LCST, ELPs phase separate, forming coacervates. They provide an excellent structural framework for bioactive peptides and their intrinsic biophysical characteristics allow for simple production with high yields and purity making them an ideal biomaterial for cell adhesion applications.Support or Funding InformationThis research is sponsored by the Japan Society for the Promotion of Science (JSPS), PE18045.
Treatment with Momordica charantia Fractions Disrupts Energy Metabolism and Elevated Reactive Oxygen Species’ Generation in Human Lung and Breast Cancer Cell Lines
Authors: Ehigie Adeola Folasade , Wei Peng , Wei Taotao , Yan Xiyun , Ojeniyi Fiyinfoluwa Demilade , Ehigie Leonard Ona 
Published: April 2020

The mechanism of action of mitochondrial targeted anti‐cancer drugs relies on their ability to disrupt the energy producing systems of cancer cell mitochondria. Momordica charantia (MC), an acclaimed anticancer plant in folk medicine, is here investigated for the probable mechanism by which it kills cancer cells. The ethyl acetate (D3), n‐hexane (D4) and dichloromethane (D5) fractions were obtained from the Crude Water‐Soluble extract (CWSE) of MC via solvent partitioning and then administered on MDA‐MB 436 and A549 cells at 100 and 125 μg/mL for 24 h. Cytofluorometric techniques, using TMRM and H2DCF‐DA as fluorescent probes were employed to assess the effect of MC on mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) generation respectively, while ATP levels were measured using the luciferin‐luciferase reaction. Mean respiring cell numbers obtained from Dot graphs and Histograms were further analyzed, yielding significant (P<0.05) MMP depressions only at 125μg/mL in both MDA‐MB 436 (43.08±2.33) and A549 (24.84±2.51) cells treated with D3 in comparison with untreated cells (175.8±6.13 and 131.7±11.21 respectively). In the same vein, only MDA‐MB 436 cells treated with D3 and D5 at 100 and 125 μg/mL significantly elevated ROS levels in comparison with untreated (100.0±2.16) cells; the highest generation being 332.1±3.20, seen in D3‐treated cells at 125 μg/mL. Furthermore, all fractions (D3‐D5) tested on A549 cells at 100μg/mL and 125μg/mL generated significantly higher levels of ROS compared with the untreated (42±4.50) group. The highest observed levels of generated ROS were seen at 125μg/mL in the D5‐treated cells (536±4.00). For intracellular ATP generation, significant (P<0.05) depletion was observed in both MDA‐MB 436 cells [only at 125μg/mL of D3 (80.63±4.70) compared with the control group (106.7±5.69)] and A549 cells [D3 at 125 μg/mL (43.68±0.13) < D5 at 125 μg/mL (52.56±4.16) < D3 at 100 μg/mL (59.84±5.39) compared with the control group (98.57±0.66)]. On the whole, this study suggests that the tested fractions of MC induced a mitochondrial‐dependent cell death in selected cancer cell lines.Support or Funding InformationOrganization for Women in Science for the Developing World (OWSD) and Swedish International Development Cooperation Agency (SIDA)
Tamarindus indica Fruit Pulp Restores Reproductive Function in Sodium Fluoride Administered Rats
Authors: Ajani Emmanuel Oladipo , Usman Lutfat Abimbola 
Published: April 2020

BackgroundInfertility is often associated with increased depression and stigmatization of infertile individuals. Although there exists in Africa, fertility inducing herbal condiments, however the pharmacology of most of these agents are unknown.ObjectiveThis study investigated the prophylactic and ameliorative effect of ethanolic fruit pulp extract of Tamarindus indica on sodium fluoride induced reproductive dysfunction.MethodologyT. indica fruit pulp was extracted in ethanol by maceration. The filtrate was thereafter oven dried at 60°C. Seventy rats of both sexes weighing 120 ± 5 g randomized into 7 groups of 10 rats each were used for the study. Group 1 was placed on placebo. Group 2 and 3 were first placed on 200 and 400 mg/kg of the extract respectively for 4 weeks followed by concomitant administration 20 mg/kg NaF. Groups 4, 5 and 6 were initially administered with 20 mg/kg NaF for 45 days followed by 4 weeks’ administration of 200 and 400 mg/kg T. indica (4 and 5) and 50 mg/kg clomid (group 6).ResultThe sperm function indices showed that administration of 400 mg/kg extract significantly increased sperm count and restore the sperm motility, morphology and viability to values not different from the control. Follicle stimulating hormone and progesterone also compared favorably with the control. The leutinizing hormone, testosterone, and oestrogen at 400 mg/kg dose were significantly raised. The histopathological analysis of the testes of rats pre‐etreated with the extract showed the presence of large interstitial space in the seminiferous tubules while a relatively normal histomorphological representation was seen in rats treated with the extract after sexual dysfunction induction.ConclusionOur results indicate that administration of the fruit pulp extract of Tamarindus indica at a dose of 400 mg/kg body weight is efficacious for the management of infertility.Support or Funding InformationPersonall
IN VITRO EVALUATION OF THE CYTOTOXIC POTENTIAL OF Securidaca longepedunculata ON HUMAN BREAST ADENOCARCINOMA (MCF‐7) CELLS
Authors: Minari Joseph , Oyedele Olaoye , Okpuzor Joy , Aja Joyce , Adewole Richard 
Published: April 2020

The prevalence of cancer has increased interest in the role of complementary and alternative medicine, employing the use of plant products. Securidaca longepedunculata (SL) serves as a major component of anticancer decoctions in Nigeria. The present study evaluates the cytotoxic potential of the crude extract and fractions of SL root bark against human breast adenocarcinoma (MCF‐7) cell line. The root back of SL was pulverized and extracted with 80% methanol to yield a crude extract which was then submitted to liquid–liquid fractionation with dichloromethane (DCM) and butanol (BUT). The extract and fractions were subjected to Gas Chromatography‐Mass Spectrometry (GC‐MS). MCF‐7 cell line was treated with graded concentrations (9.77 – 5000 μg ml−1) of the aqueous methanol crude extract as well as the DCM and BUT fractions for 24 or 48 hours. Cell viability was thereafter measured by XTT proliferation assay. The activity of the aqueous methanolic crude extract was found to be both dose‐dependent and time independent with IC50 values of 2241 μg ml−1 and 1808 μg ml−1 after 24 and 48 hours respectively. However, the DCM and BUT fractions were significantly (p < 0.05) cytotoxic with IC50 values of 86.27 μg ml−1 and 12.08 μg ml−1 respectively after 48 hours of treatment. The study lends credence that BUT fractions of SL may contain lead agent with anticancer properties and that informed its use in alternative medicine.Support or Funding InformationSELF SUPPORT
Diet‐derived gallated catechins prevent TGF‐b‐mediated epithelial‐mesenchymal transition, cell migration and vasculogenic mimicry in chemosensitive ES‐2 ovarian cancer cells
Authors: Sicard Audrey-Ann , Annabi Borhane 
Published: April 2020

BackgroundTransforming growth factor (TGF)‐β triggers ovarian cancer metastasis through epithelial‐mesenchymal transition (EMT). Whereas drug design strategies targeting the TGF‐β signaling pathway have been envisioned, the anti‐TGF structure:function aspect of chemopreventive diet‐derived catechins remains unexplored.MethodsWe assessed the effects of eight catechins on TGF‐β‐mediated cell migration using real‐time exCELLigence, induction of EMT biomarkers by RT‐qPCR and immunoblotting, and in vitro vasculogenic mimicry (VM) by 3D‐Matrigel cultures, a process partly regulated by EMT‐related transcription factors.ResultsTGF‐β‐mediated phosphorylation of Smad‐3 and p38 signaling intermediates was more effective in a chemosensitive ES‐2 ovarian cancer cell line but was inoperative in cis‐platinum‐ and adriamycin‐chemoresistant SKOV‐3 ovarian cancer cells. Increases in cell migration and in gene/protein expression of EMT biomarkers Fibronectin, Snail, and Slug were observed in ES‐2 cells. When VM was assessed in ES‐2 cells, 3D capillary‐like structures were formed and increases in EMT biomarkers found. Catechins bearing the galloyl moiety (CG, ECG, GCG, and EGCG) exerted potent inhibition of TGF‐β‐induced cell migration as well as EMT, and inhibited VM, in part through inhibition of Snail and matrix metalloproteinase‐2 secretion.ConclusionsOur data suggest that diet‐derived catechins exhibit chemopreventive properties that circumvent the TGF‐β‐mediated signaling which contributes to the ovarian cancer metastatic phenotype.Support or Funding InformationUQAM Foundation and Chaire in Cancer Prevention and Treatment
The effect of tamoxifen on gender differences in unilateral ureteral obstructed rats and human kidney slices
Authors: Tingskov Stine Julie , Jensen Michael Schou , Nørregaard Rikke 
Published: April 2020

BACKGROUNDRenal fibrosis is regarded as the most damaging process in chronic kidney disease (CKD) and it is the most consistent predictor of irreversible loss of renal function. To date, there is no good treatment for renal fibrosis. Interestingly, females are less prone to CKD, which might be due to estrogen. We have previously shown that tamoxifen (TAM), a selective estrogen receptor modulator, plays a role for regulation of renal water and salt balance. In this study, we will investigate the effect of tamoxifen on a well‐established in vivo model using unilateral ureteral obstruction (UUO)‐induced fibrosis in female, male and ovariectomized (OVX) female rats. Furthermore, a recently develop model of human renal fibrosis was used to study multicellular pathological processes in human tissue.METHODSFibrosis was induced by 7 days of UUO. Female OVX rats had both their ovaries removed two weeks before UUO. Tamoxifen (25 mg/kg for female rats and 50 mg/kg for male rats; a higher dose of tamoxifen was lethal for female rats) was given by oral gavage starting 5 days before UUO. Tamoxifen treatment was continued for 7 days after UUO. Human precision‐cut kidney slices were exposed to TGF‐β for 48h in order to make the tissue fibrotic and afterwards treated with tamoxifen for 6h. Histological changes were examined by HE staining. Expression of α‐smooth muscle actin, fibronectin, and TGF‐β were evaluated by immunohistochemistry, quantitative PCR, and western blot analysis.RESULTSRenal fibrosis was increased after UUO in both male, female and OVX rats. Tamoxifen treatment reduced fibronectin, α‐SMA and TGF‐β gene expression in female UUO rats, but not in male and in OVX rats subjected to UUO. Yet, tamoxifen significantly reduced the protein level of fibrotic markers and the kidney weight in response to UUO in both female as well as OVX and male rats. Furthermore, our data demonstrated that sham operated female and female OVX rats had a higher plasma creatinine level compared to male rats. On the other hand, TAM treated male rats subjected to UUO had a significantly increased urea plasma level, which was not observed in female and female OVX rats. Importantly, treatment with tamoxifen has a tendency to reduce fibronectin in human kidney slices exposed to TGF‐β from women but not in men.CONCLUSIONThese findings indicate that tamoxifen reduces UUO‐induced fibrosis in both female, female OVX and male rats, but have different effects on females and males when it comes to gene levels of fibrotic markers and plasma data. Furthermore, our finding provide preclinical evidence that treatment with tamoxifen affects men and women differently and the concentration of the drug must be adapted to the gender.Support or Funding InformationThis study was kindly supported by Karen Elise Jensen Foundation, the Danish Council for Independent Research, Aarhus University Foundation and the AP Møller Foundation.
Arrestin‐3 interacts with Parkin mutants linked to Parkinson’s disease
Authors: Zheng Chen , Perry Nicole A , Vishnivetskiy Sergey A , Berndt Sandra , Gurevich Eugenia V , Gurevich Vsevolod V 
Published: April 2020

Arrestins are multifunctional proteins that were originally identified as suppressors of G protein‐coupled receptors signaling. Later arrestins were found to serve as adapter molecules that bind numerous signaling proteins, such as protein kinases, ubiquitin ligases, deubiquitinases, etc. Here, we showed the direct interaction with arrestin‐3 and parkin, which is a ubiquitin E3 ligase that covalently attaches ubiquitin to specific substrates, including mitochondrial proteins. Parkin is essential for mitophagy – the removal of damaged mitochondria. Many mutations in parkin are linked to familial or sporadic Parkinson’s disease. We identified several binding hot spots on both arrestin‐3 and Parkin using high throughput peptide arrays. Some pathogenic parkin mutations map to these arrestin‐binding hot spots. One of the pathogenic mutations, R275W, significantly increased parkin binding to arrestin‐3 in cells, as compared to wild type parkin. Our data indicate that arrestin‐3 may participate in parkin‐mediated physiological processes that likely contribute to Parkinson’s disease.Support or Funding InformationThis work was supported by grant R35 GM122491 from the National Institutes of Health to VVG.
The acquisition of positional information in the developing cochlea
Authors: Munnamalai Vidhya , Young Caryl , Thompson Matthew , Fister Alexandra , Umulis David , Fekete Donna 
Published: April 2020

The sensory epithelium of the mammalian murine auditory organ, the organ of Corti, is an exquisite example of how specialized asymmetry dictates auditory function. Across the radial axis, the organ consists of three rows of outer hair cells to one row of inner hair cells, which are innervated by efferent and afferent neurons. The mechanosensory inner hair cells that reside in the neural compartment detect sound whereas the outer hair cells in the abneural compartment modulate sound levels. A complex combination of spatio‐temporal signaling inputs across the radial axis endows cells with positional information to take on specific cell fates.The Wnt and Bmp pathways are two potential signaling mechanisms that influence this radial asymmetry in an opposing manner. We analyzed the requirement of the Bmp pathway for setting up cochlear patterning. Smad4 cKOs showed a loss of the abneural compartment. Prolonged Bmp inhibition decreased the formation of the abneural compartment and outer hair cells. Bmp was required for determining the abneural boundary of the sensory domain. Immediately after hair cell differentiation, Bmp signaling was no longer required. Bmp is necessary, but not sufficient for patterning the entire epithelium. Activation of the Wnt pathway leads to a selective expansion of the neural compartment and inner hair cells. This selective neural expansion was also conserved in the chicken cochlea. Signaling crosstalk of these pathways in a spatiotemporal manner is necessary for proper patterning of the organ of Corti.Support or Funding Information
R21DC016376R01DC002756R01GM132501