Influence of Monovalent and Divalent Ions in the Conformational Change of Caspase-Cleaved Par-4 (cl-Par-4) Tumor Suppressor Protein

Prostate apoptosis response-4 (Par-4) is a pro-apoptotic tumor suppressor protein. We have shown that this 38 kDa full-length Par-4 (Fl-Par-4) protein is predominantly intrinsically disordered in vitro. In vivo, Par-4 is cleaved by caspase-3 at Asp-131 to generate a 24 kDa functionally active cleaved Par-4 (cl-Par-4) fragment. The cl-Par-4 protein inhibits the NF-κB-mediated cell survival pathway and causes selective apoptosis in various tumor cells. Our laboratory is interested in how the disorder-order balance within Fl-Par-4 and cl-Par-4 may be related to the balance between cell survival and cell death. Currently, we are using biophysical techniques such as circular dichroism (CD) spectroscopy, dynamic light scattering (DLS), UV-vis spectroscopy and SDS-PAGE to characterize the structure of cl-Par-4 in the presence of various concentrations of monovalent and divalent ions, in order to shed light on the possible ion-specific role of cl-Par-4 in inducing structure and self-association of this protein. Results show that effects on cl-Par-4 conformation are ion-specific, and effects of divalent cations are considerably more pronounced than effects from monovalent cations. We have also found that the anion moiety of a salt possesses almost negligible influence.https://digitalcommons.odu.edu/gradposters2021_sciences/1000/thumbnail.jp

Structual Analysis of the cl-Par-4 Tumor Suppressor as a Function of Ionic Environment

Prostate apoptosis response-4 (Par-4) is a proapoptotic tumor suppressor protein that has been linked to a large number of cancers. This 38 kilodalton (kDa) protein has been shown to be predominantly intrinsically disordered in vitro. In vivo, Par-4 is cleaved by caspase-3 at Asp-131 to generate the 25 kDa functionally active cleaved Par-4 protein (cl-Par-4) that inhibits NF-κB-mediated cell survival pathways and causes selective apoptosis in tumor cells. Here, we have employed circular dichroism (CD) spectroscopy and dynamic light scattering (DLS) to assess the effects of various monovalent and divalent salts upon the conformation of cl-Par-4 in vitro. We have previously shown that high levels of sodium can induce the cl-Par-4 fragment to form highly compact, highly helical tetramers in vitro. Spectral characteristics suggest that most or at least much of the helical content in these tetramers are non-coiled coils. Here, we have shown that potassium produces a similar effect as was previously reported for sodium and that magnesium salts also produce a similar conformation effect, but at an approximately five times lower ionic concentration. We have also shown that anion identity has far less influence than does cation identity. The degree of helicity induced by each of these salts suggests that the “Selective for Apoptosis in Cancer cells” (SAC) domain—the region of Par-4 that is most indispensable for its apoptotic function—is likely to be helical in cl-Par-4 under the studied high salt conditions. Furthermore, we have shown that under medium-strength ionic conditions, a combination of high molecular weight aggregates and smaller particles form and that the smaller particles are also highly helical, resembling at least in secondary structure, the tetramers found at high salt

How to prioritize patients and redesign care to safely resume planned surgery during the COVID-19 pandemic.

AIMS: Restarting planned surgery during the COVID-19 pandemic is a clinical and societal priority, but it is unknown whether it can be done safely and include high-risk or complex cases. We developed a Surgical Prioritization and Allocation Guide (SPAG). Here, we validate its effectiveness and safety in COVID-free sites. METHODS: A multidisciplinary surgical prioritization committee developed the SPAG, incorporating procedural urgency, shared decision-making, patient safety, and biopsychosocial factors; and applied it to 1,142 adult patients awaiting orthopaedic surgery. Patients were stratified into four priority groups and underwent surgery at three COVID-free sites, including one with access to a high dependency unit (HDU) or intensive care unit (ICU) and specialist resources. Safety was assessed by the number of patients requiring inpatient postoperative HDU/ICU admission, contracting COVID-19 within 14 days postoperatively, and mortality within 30 days postoperatively. RESULTS: A total of 1,142 patients were included, 47 declined surgery, and 110 were deemed high-risk or requiring specialist resources. In the ten-week study period, 28 high-risk patients underwent surgery, during which 68% (13/19) of Priority 2 (P2, surgery within one month) patients underwent surgery, and 15% (3/20) of P3 ( three months) groups. Of the 1,032 low-risk patients, 322 patients underwent surgery. Overall, 21 P3 and P4 patients were expedited to 'Urgent' based on biopsychosocial factors identified by the SPAG. During the study period, 91% (19/21) of the Urgent group, 52% (49/95) of P2, 36% (70/196) of P3, and 26% (184/720) of P4 underwent surgery. No patients died or were admitted to HDU/ICU, or contracted COVID-19. CONCLUSION: Our widely generalizable model enabled the restart of planned surgery during the COVID-19 pandemic, without compromising patient safety or excluding high-risk or complex cases. Patients classified as Urgent or P2 were most likely to undergo surgery, including those deemed high-risk. This model, which includes assessment of biopsychosocial factors alongside disease severity, can assist in equitably prioritizing the substantial list of patients now awaiting planned orthopaedic surgery worldwide. Cite this article: Bone Jt Open 2021;2(2):134-140

NC2213: a novel methionine aminopeptidase 2 inhibitor in human colon cancer HT29 cells

Methionine aminopeptidase 2 (MetAP2) is a bifunctional protein that plays a critical role in the regulation of post-translational processing and protein synthesis. MetAP2 is overexpressed in human colon cancer. In this report we screened various MetAP2 inhibitors and treated HT29 cells with various concentrations of compounds. We evaluated the expression of MetAP2 and pp60c-src expressions in HT29 cells. In addition we also carried out the cell proliferation and cell cycle analysis in the MetAP2 inhibitor-treated HT29 cells. The cell cycle analysis of HT29 treated with 1.0 μM of NC2213 showed an arrest in the G2 phase followed by an induction in the percentage of cells undergoing apoptosis in the sub-G1 phase. Western blot analysis revealed that the MetAP2 expression was dose-dependently decreased when the HT29 cells were treated with the 3,5-bis(benzylidene)-4-piperidone derivative (NC2213). In addition, phosphorylation of Src, a myristoylated oncoprotein was significantly decreased by 1.0 μM of NC2213 as revealed by Western blot analysis. Furthermore, NC2213 also inhibits the expression of pp60c-src in HT29 cells. Interestingly, this compound also inhibits the phosphorylation at Tyr416 of pp60c-src while increasing the phosphorylation at Tyr527 of pp60c-src. NC2213 inhibits the growth of HT29 cells by inducing apoptosis and might be useful for the treatment of human colon cancer

Successful transportation of wild stock of endangered "Thooli" (Labeo dussumieri) spawners without anaesthetics

Several problems are encountered during transportation of fishes from wild to captivity. The note describes successful transportation of wild caught endangered Labeo dussumieri spawners over distances below 100 km without anesthesia or sophisticated transporting devices which is of immense help in the gene banking programme of endangered species

National Strategic Plan for Aquatic Exotics and Quarantine

India is one of the richest countries in the world in terms of fish genetic wealth. But since 19th century, there have been introduction of over 350 fish species into the country for sport fishery, aquaculture, ornamental fishery and insect control purposes. Translocation of species across political boundaries of nations has been a centuries old phenomenon. The transboundary movement of species for culture or for aquarium purposes has witnessed a spurt during the past century and is expected to get accelerated in the coming years. With the country's attempt to enhance the yield and with the demand of the fast growing ornamental fish trade, the fishery industry will be keen to import more promising strains/ species. Some of the introduced species have added to the fish production, but some species proved to be invasive in our open waters, affecting the native species. The potential and real impacts of introducing new species into the aquatic ecosystem have been major issues debated worldwide. The benefits of introduced species are usually immediate and for short duration whereas the adverse impacts of exotics become evident only after a long time. The long-term impacts of the introduced species have not been evaluated till date in our country. Most conclusions regarding beneficial or adverse impacts of exotics are based on individual perceptions and are usually not supported by scientific data. Pre-introductory data to evaluate the impact of introduced species should be taken into consideration before drawing conclusions. In this context, it is necessary to provide feasible guidelines for the import of new species or strains depending on the purpose of import. Under the Convention on Biodiversity (CBD), India has the moral and legal obligations to conserve its biodiversity. There is need to have adequate safeguards to protect the native species from the onslaught of the introduced ones. At the same time the nation must also ensure that the fish yield is enhanced and international trade, especially in ornamental fishery expands. Industry and traders should be allowed to develop and expand the trade for the economic benefit of the country. In India, the policy on introduction of exotic aquatic organisms is not clearly defined. Moreover, there is no policy to tackle exotics already introduced both legally and illegally

DNA sequence information resolves taxonomic ambiguity of the common mud crab species (Genus Scylla) in Indian waters

For several years, mud crabs of genus Scylla have been misidentified owing to their high morphological plasticity and the absence of distinct morphological diagnostic characters. The taxonomic confusion of genus Scylla de Haan is considered to be a primary constraint to the development of aquaculture. Although genus Scylla was revised using morphological and genetic characteristics, taxonomy of Scylla species occurring in India is still not clear. In this study, partial sequences of two mitochondrial genes, 16S rRNA and CO1 (Cytochrome C oxidase subunit I) in populations of Scylla spp. obtained from eleven locations along the Indian coast were used to differentiate and resolve taxonomical ambiguity of the mud crab species in India. The sequences were compared with previously published sequences of Scylla spp. Both trees generated based on 16S rRNA and CO1 indicated that all S. tranquebarica morphotypes obtained during this study and S. tranquebarica sequences submitted previously from Indian waters reciprocally monophyletic with reference sequence of S. serrata. Both sequence data and morphological characters revealed that the species S. serrata (Forskal) is the most abundant followed by S. olivacea. Further, the 16S rRNA and COI haplotypes of Indian S. tranquebarica obtained in the study significantly differed with the known S. tranquebarica by 6.7% and 10.6% respectively whereas it differed with known S. serrata by 0.0–0.7% only, a difference that was not statistically significant. From these studies it is clear that ‘‘S. tranquebarica’’ commonly reported from India should be S. serrata (Forskal)

Low Genetic Differentiation in the Populations of the Malabar Carp Labeo dussumieri as revealed by Allozymes, Microsatellites and RAPD

The population structure of Labeo dussumieri, an endangered and endemic cyprinid from three riverine locations in the Western Ghats, India was investigated using allozyme, microsatellite and RAPD markers. L. dussumieri samples were obtained from Meenachil, Manimala and Pamba River basins, Kerala. Fourteen (46.7%) out of 30 allozyme loci, seven microsatellite loci and 12 RAPD Operon decamers gave polymorphic pattern. Six allozyme loci (AAT-2*, EST-4*, GLDH*, GPI-2*, G6PDH* and LDH-2*) and three microsatellite loci (LdussG1, MFW19 and Bgon22) exhibited consistent significant deviation from Hardy-Weinberg Equilibrium expectations in different populations after probability level (P<0.05) was adjusted for sequential Bonferroni correction. All the three marker types demonstrated concordant results and various estimates revealed genetic variability within the subpopulations but surprisingly low level (= 0.0034 to 0.0081) of genetic differentiation among L. dussumieri from different river samples. AMOVA analysis also indicated low differentiation among subpopulations. No evidence for a recent genetic bottleneck was observed in L. dussumieri populations based on allozyme and microsatellite data set analysis. Meenachil, Manimala and Pamba Rivers open in to the southern end of Vembanad Lake in Kerala and are connected to each other in the lower reaches through an extensive network of natural canals. Common ancestry in the prehistoric period; and possible mixing of fish populations resulting in high gene flow across the rivers through the lake and interconnecting canals could have been responsible for the lack of significant allelic heterogeneity among the L. dussumieri populations. The stocks from the three rivers do not require different management strategies and for propagation assisted river ranching programme of this species, large effective breeding population can be developed by mixing individuals from three river

Increasing incidence of spondylodiscitis in England: an analysis of the national health service (NHS) hospital episode statistics from 2012 to 2021

Background Spondylodiscitis is a potentially life-threatening infection of the intervertebral disk and adjacent vertebral bodies, with a mortality rate of 2–20%. Given the aging population, the increase in immunosuppression, and intravenous drug use in England, the incidence of spondylodiscitis is postulated to be increasing; however, the exact epidemiological trend in England remains unknown. Objective The Hospital Episode Statistics (HES) database contains details of all secondary care admissions across NHS hospitals in England. This study aimed to use HES data to characterise the annual activity and longitudinal change of spondylodiscitis in England. Methods The HES database was interrogated for all cases of spondylodiscitis between 2012 and 2019. Data for the length of stay, waiting time, age-stratified admissions, and ‘Finished Consultant Episodes’ (FCEs), which correspond to a patient's hospital care under a lead clinician, were analysed. Results In total, 43135 FCEs for spondylodiscitis were identified between 2012 and 2022, of which 97.1% were adults. Overall admissions for spondylodiscitis have risen from 3 per 100,000 population in 2012/13 to 4.4 per 100,000 population in 2020/21. Similarly, FCEs have increased from 5.8 to 10.3 per 100,000 population, in 2012–2013 and 2020/21 respectively. The highest increase in admissions from 2012 to 2021 was recorded for those aged 70–74 (117% increase) and aged 75-59 (133% increase), among those of working age for those aged 60–64 years (91% increase). Conclusion Population-adjusted admissions for spondylodiscitis in England have risen by 44% between 2012 and 2021. Healthcare policymakers and providers must acknowledge the increasing burden of spondylodiscitis and make spondylodiscitis a research priority

Acidity in Vembanad Lake causes fish mortality

The sudden changes of variations in the environmental and climatic patterns in an area always cause certain undesirable and unforeseen events as well as problems. Immediately after the first monsoon rains, in the third week of June, an instance of mass mortality of fishes and clams was reported from the Vembanad Lake. The report indicated that large scale mortality of several groups of aquatic organisms were occurring mainly in the southern half of the lake. The unusual phenomena might have been caused by soil acidity. Hydrogen-ion concentration of the water showed that it was in acid range. The data revealed that the effect of pH reduction seemingly influenced the eco-system only up to the middle of the lake and coming towards the mouth of the estuary the effect appeared to have got neutralized, probably due to the regular tidal action. The reason for the persisting low pH could be that the river water input into the lake after several spells of monsoons showers has not been sufficient to flush the acids produced at the water soil interface continuing for several weeks as the monsoon this season had been weak and halting in the initial period