Exp Biol Med Research Articles

100 years flashback

100 years flashback

ODP417 Kisspeptin directly stimulates estrogen and progesterone secretion from human granulosa cells.

Abstract Kisspeptin is a peptide hormone that has a central role to stimulate gonadotropin-releasing hormone secretion from the hypothalamus. Furthermore, it was shown to have peripheral roles on enhancing CYP19 (aromatase) expression and aromatase secretion from human granulosa cells (1). The objectives of this study were to determine direct effects of kisspeptin treatment on estrogen and progesterone secretion from human granulosa-like tumor cell line (KGN). KGN cells (n=3) were treated with 10 nM follicle stimulating hormone (FSH); 100, 1,000, and 10,000 nM kisspeptin; and FSH together with different doses of kisspeptin for 24 hours. Estrogen and progesterone concentrations in supernatant were measured by enzyme immunoassay kits. The results showed that FSH treatment alone (P<0. 05) and FSH together with 100 (P<0. 01), 1,000 (P<0. 001), and 10,000 (P<0. 001) nM kisspeptin increased estrogen concentrations in supernatants when compared to control. Kisspeptin treatment alone did not increase estrogen concentrations in supernatant when compared to control. Interestingly, kisspeptin treatment at 1,000 (P<0. 01) and 10,000 (P<0. 01) nM together with FSH enhanced estrogen concentrations in supernatant when compared to FSH treatment alone. Furthermore, FSH treatment alone (P<0. 05) and FSH together with kisspeptin at doses of 1,000 (P<0. 001) and 10,000 (P<0. 05) nM increased progesterone concentrations in supernatant when compared to control. All doses of kisspeptin did not increase progesterone concentrations in supernatant when compared to control. Only 1,000 nM kisspeptin together with FSH enhanced progesterone secretion in supernatant when compared to FSH alone (P<0. 01). Taken together, FSH stimulated estrogen and progesterone secretion from human granulosa cells and kisspeptin enhanced these effects in addition to the effects of FSH alone. Further studies are required to determine whether kisspeptin treatment can restore the functions of unresponsive or partially responsive granulosa cells of aging humans to secrete estrogen and progesterone as well as kisspeptin is useful in augmenting infertility treatment. Reference: (1) Qin L, Sitticharoon C, Petyim S, Keadkraichaiwat I, Sririwichitchai R, Maikeaw P, et al. Exp Biol Med (Maywood). 2021;246(8): 996-1010. Presentation: No date and time listed

Aldehyde dehydrogenase 2 and arrhythmogenesis.

Cardiac arrhythmia is a common cardiovascular disease that leads to considerable economic burdens and significant global public health challenges. Despite the remarkable progress made in recent decades, antiarrhythmic therapy remains suboptimal. Aldehyde dehydrogenase 2 (ALDH2), a critical detoxifying enzyme, catalyzes toxic aldehydes and protects individuals from damage caused by oxidative stress. Accumulating evidence has demonstrated that ALDH2 activation has potential antiarrhythmic benefits. The correlation between ALDH2 deficiency and arrhythmogenesis has been widely recognized. In this review, we summarize recent research on the potential role of ALDH2 activation and antiarrhythmic protection, as well as the role played by the ALDH2∗2 polymorphism (rs671) in promoting arrhythmic risk. Additionally, we discuss important new findings illustrating the use of ALDH2 activators, which may prove to be promising antiarrhythmic therapy agents.

Vitamin C and its therapeutic potential in the management of COVID19.

COVID19 has emerged as one of the worst pandemics in the history of mankind. Several vaccines have been approved by different government agencies worldwide, but data on their efficacy and safety are limited, and distribution remains a massive challenge. As per WHO, personal immunity is vital for protection against COVID19. Earlier, Vitamin C-mediated pathways have been shown to play critical role in boosting immunity attributed to its antioxidant properties. Recently, the involvement of such pathways in protection against COVID19 has been suggested. The controlled doses of Vitamin C administered through intravenous (IV) injections are being studied for determining its role in the prognosis of COVID19. In this article, we have discussed the potential role of Vitamin C in the management in COVID19 patients and presented recent clinical trials data. Additionally, we have elaborated the possibility of administering Vitamin C through inhalers in order to achieve local high concentration and the challenges of such approach.

Financial insecurity among children with atopic dermatitis in the United States

Financial insecurity among children with atopic dermatitis in the United States

Commentaries on Viewpoint: The interaction between SARS-CoV-2 and ACE2 may have consequences for skeletal muscle viral susceptibility and myopathies.

Commentaries on Viewpoint: The interaction between SARS-CoV-2 and ACE2 may have consequences for skeletal muscle viral susceptibility and myopathies.

In Memoriam: Marvin A. Van Dilla: 1919-2019.

© 2020 International Society for Advancement of Cytometry.

Intravenous Injection of Muse Cells as a Potential Therapeutic Approach for Epidermolysis Bullosa

Intravenous Injection of Muse Cells as a Potential Therapeutic Approach for Epidermolysis Bullosa

Xenogeneic transplantation of human WJ-MSCs rescues mice from acute radiation syndrome via Nrf-2-dependent regeneration of damaged tissues.

There is an unmet medical need for radiation countermeasures that can be deployed for treatment of exposed individuals during ionizing radiation (IR) accidents or terrorism. Wharton's jelly mesenchymal stem cells (WJ-MSCs) from human umbilical cord have been shown to avoid allorecognition and induce a tissue-regenerating microenvironment, which makes them an attractive candidate for mitigating IR injury. We found that WJ-MSCs protected mice from a lethal dose of IR even when transplanted up to 24hours after irradiation, and a combination of WJ-MSCs and antibiotic (tetracycline) could further expand the window of protection offered by WJ-MSCs. This combinatorial approach mitigated IR-induced damage to the hematopoietic and gastrointestinal system. WJ-MSCs increased the serum concentration of the cytoprotective cytokines granulocyte colony-stimulating factor (G-CSF) and IL-6 in mice. Knockdown of G-CSF and IL-6 in WJ-MSCs before injection to lethally irradiated mice or transplantation of WJ-MSCs to lethally irradiated Nrf-2 knockout mice significantly nullified the therapeutic protective efficacy. Hence, WJ-MSCs could be a potential cell-based therapy for individuals accidentally exposed to radiation.

Corrigendum

Liu J, Fei L, Huang G-Q, Shang X-K, Liu M, Pei Z-J, Zhang Y-X. Right ventricle performances with echocardiography and 99mTc myocardial perfusion imaging in pulmonary arterial hypertension patients. Exp Biol Med 2018;243(9):754-761. In this article, the first author's affiliation was incorrect in the OnlineFirst and print versions. It should have appeared as 'Department of Nuclear Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China'. This has been corrected in the online article.

PWE-061 Dysbiosis in Patients with Bile Acid Diarrhoea (BAD) Demonstrated Using 16S RNA Gene Sequencing

Introduction The gastrointestinal tract harbours a diverse community of approximately 1014 microorganisms comprising 500 to 1000 bacterial species.1 Dysbiosis occurs when pathological imbalances in the gut bacterial community precipitate disease and has been linked to the dysmetabolism of bile acids (BAs) in the gut. 1–2% of primary BAs escape the enterohepatic circulation and undergo microbial biotransformation in the large bowel to form the secondary BAs, deoxycholic acid (DCA) and lithocholic acid (LCA).2 BA metabolites as a result of microbial transformations act as signalling molecules via the Farnesoid X receptor (FXR). Activation of the ileal FXR stimulates expression of Fibroblast Growth Factor 19 (FGF-19), which binds hepatic FGF-4 and activates JNK1/2 and ERK1/2 to impede the physiological feedback of BA synthesis and maintain a functional BA pool.3 The degree of activation of BA receptors is influenced primarily by the gut microbiota and therefore dysbiosis may result in abnormal BA modification leading to the development of gastrointestinal disease. To enhance our understanding of gut dysbiosis in patients with BAD, 16 S rRNA gene sequencing was undertaken to investigate bacterial communities from faecal samples in patients with BAD and IBS. Methods 35 patients (20 with BAD and 15 with IBS) were recruited from the nuclear medicine department at University Hospitals Coventry and Warwickshire NHS Trust after referral from the gastroenterology clinic for a SeHCAT scan to investigate chronic diarrhoea. The patients with BAD had either Type 1 with a diagnosis of Crohn’s disease, type 2 or type 3 post cholecystectomy. Stool samples were stored at -80oC within two hours of collection and then defrosted for 45 minutes at room temperature prior to analysis. 16 s RNA sequencing was undertaken by isolating DNA from stool samples using the QIAamp Fast DNA Stool Extraction kit. V3-V4 primers and extensor ready mix (Thermo scientific) were used to amplify the 16 s rRNA gene sequences from isolated metagenomic DNA. The eluted DNA was then quantified using the broad-range Qubit kit. DNA was analysed by electrophoresis on 1% agarose gels and purified post-PCR. The DNA was then denatured and sequenced on a MiSeq using the Illumina Miseq V2 2x250 bp paired end protocol. Bioinformatic statistical analysis was undertaken using QUIIME and the UPARSE pipeline. Results The rarefaction plots showed reduced diversity in BAD patients compared with IBS; p = 0.0007216. This was also confirmed using the Shannon’s diversity index. The abundance of certain taxa namely Lachnospiraceae (p = 0.00134), Rumiinococcus (p = 0.0034) and Prevotella (p = 0.0046) were reduced in patients with BAD. Conclusion We have shown that diversity in those with BAD is significantly reduced compared to those with IBS with certain phyla predominating. References 1 Gerard P. Metabolism of Cholesterol and Bile Acids by the Gut Microbiota. Pathogens 2014;3:14–24. 2. Vlahcevic ZR, Heuman DM, Hylemon PB. Physiology and pathophysiology of enterohepatic circulation of bile acids. In D. Zakim and T. Boyer, editors. Hepatology: A Textbook of Liver Disease. 1996, 3rd edition. Vol. 1. Saunders, Philadelphia, PA. 376–417 3 Tsuei J, Chau T, Mills D, et al. Bile acid dysregulation, gut dysbiosis, and gastrointestinal cancer. Exp Biol Med (Maywood). 2014 Nov;239(11):1489–504. Disclosure of Interest None Declared

The lymphotoxin β receptor is a potential therapeutic target in renal inflammation.

Accumulation of inflammatory cells in different renal compartments is a hallmark of progressive kidney diseases including glomerulonephritis (GN). Lymphotoxin β receptor (LTβR) signaling is crucial for the formation of lymphoid tissue, and inhibition of LTβR signaling has ameliorated several non-renal inflammatory models. Therefore, we tested whether LTβR signaling could also have a role in renal injury. Renal biopsies from patients with GN were found to express both LTα and LTβ ligands, as well as LTβR. The LTβR protein and mRNA were localized to tubular epithelial cells, parietal epithelial cells, crescents, and cells of the glomerular tuft, whereas LTβ was found on lymphocytes and tubular epithelial cells. Human tubular epithelial cells, mesangial cells, and mouse parietal epithelial cells expressed both LTα and LTβ mRNA upon stimulation with TNF in vitro. Several chemokine mRNAs and proteins were expressed in response to LTβR signaling. Importantly, in a murine lupus model, LTβR blockade improved renal function without the reduction of serum autoantibody titers or glomerular immune complex deposition. Thus, a preclinical mouse model and human studies strongly suggest that LTβR signaling is involved in renal injury and may be a suitable therapeutic target in renal diseases.

Intermittent Hypoxia Conditioning (IHC) and Delta Opioid Receptor (DOR) Expression in Canine Atrium

Extended hypoxia exposures make hearts vulnerable to damage and reportedly suppress DOR expression in cell models resulting in cell damage. However, 20 days of limited daily IHC produced a robust resistance to ischemia reperfusion injury in canine hearts (Exp Biol Med 2004;229:806‐812). DOR blockade with naltrindole during daily IHC abrogates this cardioprotection. Since DOR stimulation enhances cardioprotective cholinergic systems this study evaluated whether controlled IHC alters DOR, cholinergic and/or adrenergic systems to augment cholinergic control of the heart during ischemia reperfusion. Dogs underwent normoxic sham conditioning, IHC [5‐8 cycles of 5‐10 min hypoxia (9.5‐10% FIO2) + 4 min normoxia], or IHC + daily sc naltrindole before IHC, for 20 d. On day 21, atrial tissue was biopsied for immunoblot analysis of proteins. Relative to sham values, after IHC DORs trended lower but after IHC + naltrindole were higher relative to sham (p<0.05) and IHC (p<0.01). Since DOR blockade abrogates IHC mediated protection, these data suggest that DOR stimulation during IHC down‐regulates DOR with a compensatory increase after DOR blockade. Atrial tyrosine hydroxylase was largely unchanged following IHC, while multiple markers of cholinergic function were mixed, resulting in a potential increase in cholinergic influence.

Promoter Targeting RNAs: Unexpected Contributors to the Control of HIV-1 Transcription

In spite of prolonged and intensive treatment with combined antiretroviral therapy (cART), which efficiently suppresses plasma viremia, the integrated provirus of HIV-1 persists in resting memory CD4+ T cells as latent infection. Treatment with cART does not substantially reduce the burden of latent infection. Once cART is ceased, HIV-1 replication recrudesces from these reservoirs in the overwhelming majority of patients. There is increasing evidence supporting a role for noncoding RNAs (ncRNA), including microRNAs (miRNAs), antisense (as)RNAs, and short interfering (si)RNA in the regulation of HIV-1 transcription. This appears to be mediated by interaction with the HIV-1 promoter region. Viral miRNAs have the potential to act as positive or negative regulators of HIV transcription. Moreover, inhibition of virally encoded long-asRNA can induce positive transcriptional regulation, while antisense strands of siRNA targeting the NF-κB region suppress viral transcription. An in-depth understanding of the interaction between ncRNAs and the HIV-1 U3 promoter region may lead to new approaches for the control of HIV reservoirs. This review focuses on promoter associated ncRNAs, with particular emphasis on their role in determining whether HIV-1 establishes active or latent infection.

AB0323 Vitamin D receptor gene polymorphisms in rheumatoid arthritis

BackgroundThe etiology of rheumatoid arthritis (R.A.), an autoimmune, polygenic, inflammatory disease, has not yet been clarified despite the intense research1. In addition to its more commonly known metabolic effects such...

AB0657 25 hydroxy D vitamin status in patients with primary sjÖgren’s disease

BackgroundPrimary Sjogren’s syndrome (pSS) is characterised by diminished exocine glandular functions caused by autoimmune epithelitis due to dysregulated immune response. Low vitamin D level might be one of its etiologic...

AB0211 Glutathione s-transferases and cytochrome p450 enzymes polymorphisms as susceptibility factors to systemic lupus erythematosus in southern brazilian patients

BackgroundSystemic lupus erythematosus (SLE) is an autoimmune chronic inflammatory disease that presents a variety of clinical manifestationsand immunological abnormalities.There is evidence that immunological, environmental, hormonal and genetic factors may contribute...

A Rationally Designed A34R Mutant Oncolytic Poxvirus: Improved Efficacy in Peritoneal Carcinomatosis

Oncolytic poxviruses have demonstrated initial promising results in patients with cancer in clinical trials, yet further improvements are needed. It has been shown that a single point mutation in the A34R gene resulted in the production of more total progeny virus and more extracellular enveloped virus (EEV), a form that can be immune-evasive and with enhanced spread. We have genetically engineered a new oncolytic poxvirus (designated vA34R) by incorporating this mutated A34R gene into a viral backbone (vvDD) which was designed for tumor-selective replication. This rationally designed virus can evade neutralization from antipoxvirus antibodies and is highly cytotoxic to cancer cells. It demonstrates improved spread and increased replication within the peritoneal cavity resulting in improved antitumor effects in a peritoneal carcinomatosis (PC) model of MC38 colon cancer. Impressively, after carrier cell-mediated delivery in the preimmunized host, vA34R displayed high replication in tumor nodules yet low accumulation in normal tissues thus enhancing the therapeutic index leading to 70% long-term cures. These results demonstrate that vA34R gains an enhanced therapeutic index for PC via immune evasion, increased spread, and production of more progeny virus. Thus, vA34R may be a potent oncolytic virus (OV) for patients with PC, even after prior exposure to vaccinia virus (VV).

Control of Cell Identity in Pancreas Development and Regeneration

The endocrine and exocrine cells in the adult pancreas are not static, but can change their differentiation state in response to injury or stress. This concept of cells in flux means that there may be ways to generate certain types of cells (such as insulin-producing β-cells) and prevent formation of others (such as transformed neoplastic cells). We review different aspects of cell identity in the pancreas, discussing how cells achieve their identity during embryonic development and maturation, and how this identity remains plastic, even in the adult pancreas.

Fibulin-1 is Down-Regulated Through Promoter Hypermethylation and Suppresses Renal Cell Carcinoma Progression

Renal cell carcinoma is one of the most common cancers worldwide but the molecular mechanisms that underlie it are not fully understood. Fibulin-1, a multi-functional extracellular matrix protein, is involved in many types of cancer but its function in renal cell carcinoma is unclear. We investigated fibulin-1 expression and function in renal cell carcinoma. We performed real-time polymerase chain reaction, Western blot analysis and immunohistochemistry to determine fibulin-1 expression in renal cell carcinoma cells and patient tissues. Methylation specific polymerase chain reaction and quantitative sequencing were done to examine the methylation status of the FBLN1 gene promoter. Eukaryotic expression plasmid and a lentiviral vector were used to over express fibulin-1 in ACHN and 786-O renal cell carcinoma cells to investigate its function in vitro and in vivo. Fibulin-1 was significantly down-regulated in renal cell carcinoma cell lines and patient tissues. Dysregulation was associated with renal cell carcinoma progression. The FBLN1 gene promoter region was hypermethylated and its methylation status correlated with fibulin-1 expression. Fibulin-1 over expression led dramatically to decreased cell growth, enhanced tumor cell apoptosis, decreased cell motility and angiogenesis in cultured renal cell carcinoma cells and in xenograft tumors in nude mice. Fibulin-1 is down-regulated in renal cell carcinoma through promoter hypermethylation. It functions as a tumor suppressor and angiogenesis inhibitor in renal cell carcinoma.