Effect of MUC16 Blockade using the Humanized AR9.6 Antibody in Patient Derived Organoid Models of PDAC

Pancreatic Ductal Adenocarcinoma (PDAC) represents nearly 90% of all pancreatic cancer cases. 49,830 of the 62,210 patients diagnosed in 2022 are estimated to succumb to the malignancy. Early diagnosis of the disease is uncommon as most patients present with symptoms when the cancer is late-stage and metastatic. This decreases the likelihood of successful surgical resection and increases the dependency on standard of care chemotherapy leads to which is met with therapeutic resistance, demonstrated by the 5-year post-diagnosis survival rate of a mere 11.5%. Mucin-16, a heavy glycosylated transmembrane protein is overexpressed in more than 65% PDAC cases. AR9.6 is an anti-MUC16 antibody that has been recently humanized after evidence of its therapeutic potential was found in an orthotopic study utilizing the murine version of the antibody. The HuAR9.6 antibody efficiently binds MUC16 expressed on tumor cells, and can both inhibit downstream oncogenic signaling and elicit tumor killing by signaling the immune system to the tumor. In this project, we used RNA sequencing to evaluate the MUC16 mediated transcriptomic changes by using the humanized AR9.6 antibody in patient-derived organoid models of PDAC. To begin this study, organoids were developed using tumor cells from a primary PDAC tumor with a high MUC16 profile obtained from rapid autopsy patient #142 from the Rapid Autopsy Program at UNMC. The organoids were then treated in triplicates using a monoclonal antibody HuIgG as an isotype control due to its lack of specificity, and the test arm of study, HuAR9.6. These samples were treated with 40 ug/mL of the antibodies for a 24-hour period, post which RNA isolation was performed. RNA sequencing and subsequent Gene Ontology and KEGG enrichment analysis revealed a downregulation of genes involved in the Hippo signaling pathway, fat digestion and absorption and TGF-β signaling. Based on this gene expression profiling, we hypothesize that HuAR9.6 can slow tumor progression by downregulating the Hippo and TGF-β signaling pathways. In the future, we aim to robustly validate these results at the level of the proteome and assess if these results can be reproduced in multiple patient samples with the hope to translate this antibody to the clinic to be used in PDAC patients who have a high MUC16 expression.https://digitalcommons.unmc.edu/surp2022/1038/thumbnail.jp

A study to assess effectiveness of video assisted teaching programme on gastro intestinal tract assessment among student nurses of tertiary care hospital: a statistical approach

Background: The gastro intestinal is important system in the human body. Many times it is found that its uneasy for student nurses to check the patient. So, there is need to improve the available learning programmes regarding gastro intestinal tract assessment.So, it was necessary to conduct this study for educating student nurses regarding the gastro intestinal tract assessment to improve their knowledge. The objectives of this study were to assess the knowledge of student nurses regarding gastro-intestinal tract assessment and to evaluate the effectiveness of video assisted teaching programme on knowledge regarding gastro intestinal tract assessment.Methods: Material and methods used for the study is the evaluative approach; one group pre test, post test design was used. Study was conducted on sample of 70 student nurses by using convenient sampling technique. The data were collected by structured questionnaire. The data were analyzed using descriptive and inferential statistics.Results: The mean knowledge score of student nurses during the pre-test was 39.89% where as it had raised up to 72% during the post-test regarding gastro intestinal tract assessment as effectiveness of video assisted teaching programme. Therefore, the difference assessed was 32.11% between pre-test and post-test.Conclusions: There was significant difference between the pre-test knowledge level and post-test knowledge level of student nurses on gastro intestinal tract assessment. Hence health education programs and on-going teaching both can further improve the knowledge of student nurses

Supercapacitor Supported by Nickel, Cobalt and Conducting Polymer Based Materials: Design Techniques and Current Advancement

The recent advanced electronic appliances demand special high power devices with lightweight, flexible, inexpensive, and environment friendly in nature. In addition, for many industrial and automotive applications, we need energy storage systems that can store energy in a short time and deliver an intense pulse of energy for long duration. Till date the Li-ion battery is the only choice for fulfilling all our energy storage demands. However, the high cost, limited availability and non-environmental nature of electrodes and electrolyte material of Li-ion battery limits its applicability. Hence, the world demands an alternative replacement for the Li-ion battery. In this regard, the supercapacitor is one of the most emerging and potential energy storage devices. The electrode plays an important role in supercapacitors. The nickel and cobalt based oxide, hydroxides, and their composites with conducting polymer are promising and highly appreciated electrode materials for supercapacitors. This chapter covers the recent advances in supercapacitors supported by nickel, cobalt and conducting polymer based materials and their applications predominantly described in the recent literature. Recent advances are reviewed including new methods of synthesis, nanostructuring, and self-assembly using surfactant and modifiers. This chapter also covered the applications of supercapacitors in powering the light weight, flexible and wearable electronics

Graduation of the Learning Curve from Small to Smaller: Evolution of Tubular Retractors from 18 mm to 14 mm in Management of Lumbar Disc Herniation

Objective While the utility of 16 mm and 18 mm diameter tubular-retractors in the management of prolapsed intervertebral-disc is well-established, there is no published literature on the use of 14 mm tube. The aim of this study is to retrospectively analyze the feasibility and outcomes of discectomy performed through a 14 mm diameter tube in comparison with 16 mm and 18 mm diameter tubes. Methods Groups A, B and C consisting of the first 40 operated patients with symptomatic lumbar disc herniation using 18 mm, 16 mm and 14 mm tubes respectively between July-2007 and April-2018 were evaluated for VAS (leg pain) and ODI and followed up at 1 week, 6 weeks, 3 months, 6 months and 1-year intervals. Results The mean age in Groups A, B and C was 45.3 years (16-78), 47.15 years (20-78) and 42.15 years (17-65) respectively. The mean VAS in Group A, B and C improved from 7.89, 8.15 and 8.2 to 2.53, 2.4 and 2.34 respectively. The ODI in Groups A, B and C improved from 58.5, 56.4, 54.4 to 28.5, 25.6 and 22.4 respectively. The operative time and blood loss in Groups A, B and C were 106.9±44.9, 74±19.45 and 53.37±12.26 min and 54.75±21.11 mL, 47.5±11.03 mL and 43.75±6.86 mL respectively (p<0.0001). There were 8 complications in Group A, 4 in Group B and 2 in Group C. Conclusion In experienced hands there was a favourable trend with regards to operative feasibility, surgical time, surgical outcomes and complication rates with decreasing diameters of the tubular-retractor

Health Policy Implementation in Developing Nations: Challenges and Solutions

Putting health plans into action in poor countries is hard for many reasons, which makes it harder to provide good healthcare to the people who live there. Policies are often hard to put into action because of a lack of money, facilities, and skilled healthcare workers. Also, government uncertainty and corruption can make health projects less effective than they were meant to be.&nbsp; One major obstacle is the insufficient funding allocated to health programs, leading to inadequate facilities and a shortage of essential medical supplies. This financial constraint exacerbates the difficulty of attracting and retaining qualified healthcare professionals, perpetuating a cycle of suboptimal healthcare delivery.Infrastructure deficiencies, including poor road networks and limited access to remote areas, further compound implementation challenges. These obstacles impede the timely and equitable distribution of healthcare services, disproportionately affecting rural and marginalized communities.Political instability and corruption introduce an additional layer of complexity, compromising the integrity of health policy implementation. Unstable political environments often result in inconsistent policy frameworks, hindering long-term planning and sustainable healthcare improvements. Corruption erodes trust in the healthcare system, discouraging public participation and impeding the successful execution of health policies.To address these challenges, a multi-faceted approach is essential. Increased international collaboration and financial assistance can alleviate resource constraints, while targeted capacity-building initiatives can bolster the healthcare workforce. Improved infrastructure development, especially in rural areas, is crucial for ensuring widespread access to healthcare services. Additionally, fostering political stability and implementing anti-corruption measures are vital steps toward creating an enabling environment for successful health policy implementation in developing nations

Effect of Pulsed Current on TIG Weldments of Aluminium Alloy (5052) and Alloy Steel (EN24)

ABSTRACT: In this experimental work destructive and non-destructive tests have been carried out on an aluminium alloy(5052) and alloy steel of EN24 using GTAW with pulsed and non-pulsed current at different frequencies 3Hz 5Hz,&amp;7Hz.The Liquid Penetrant, radiography and tensile strength of weldments were evaluated and compared with pulsed and non-pulsed current welding at different frequencies of two materials (5052 aluminium alloy of 2.5mm thick and alloy steel EN24 of 4.5 mm thick).The aim of this experimental work is to see the effect of pulsed current on the quality of weldments. The experimental results pertaining to different pulsed current welding for the above materials using pulsed and non-pulsed current GTAW are discussed and compare

Development of Online Defect Detection System for High End Wires

Wire rods are drawn into various diameters in multi pass wire drawing machines. These drawn wires should be free from surface defects. Sometimes surface defects are observed in finished wires which can be caused due to poor wire rod surface quality or can be generated during wire drawing process. At present quality assurance is based on visual inspection. Surface quality of entire length of drawn wire cannot be ensured by the existing system causing defective wires delivered to customers. There is a need to develop and implement a low cost online defect detection system to ensure that the finished wires are free from surface defects. In this work, a new system for online inspection of thin wires has been developed. The system will be capable of detecting various types of defects like sliver, transverse crack, fin and lap discontinuities that commonly arise in the process of wire drawing based on the signature of the received signal. This paper will demonstrate the need and way forward to develop a low cost defect identification system for thin wires

FORMULATION DEVELOPMENT & EVALUATION OF EFFERVESCENT TABLET OF ALENDRONATE SODIUM WITH VITAMIN D3

Alendronate sodium is a bisphosphonates which has antiresorptive effect which is implicated in the prophylaxis and treatment of osteoporosis. The objective of this study was to formulate effervescent tablet of Alendronate sodium with Vitamin D3 against osteoporosis thereby improving patient compliance.                                                                                                                          As per revised definition proposed to US FDA, Effervescent tablet is a tablet intended to be dissolved or dispersed in water before administration.         Effervescent tablets were formulated using citric acid and sodium bicarbonate as effervescent composition by wet granulation. The drug-excipient compatibility study done by DSC &amp; FTIR analysis and it reveals absence of interaction between the drug and excipients. The flowability study of precompression blend shows good flow properties. Formulation was evaluated for weight variation, thickness, hardness, solution time, pH of solution &amp; content uniformity. All the evaluation parameters were within the limit and complies specifications as per U.S.P. &amp; B.P. From the Stability analysis may be inferred that there was no degradation and change in the formulation.                                                                                                                                                  The Effervescent tablet of Sodium Alendronate and Vitamin D3 is a new pharmaceutical formulation to be taken orally and offering a considerable advantage: avoidance of gastro-intestinal disorders, to the limits of the possible. As compared to the pure drug and marketed tablet, this formulation displayed significantly effective in the oral osteoporosis treatment in post menopausal women

Insect Cell Expression and Purification of Recombinant SARS-COV-2 Spike Proteins That Demonstrate ACE2 Binding

The COVID-19 pandemic caused by SARS-CoV-2 infection has led to socio-economic shutdowns and the loss of over 5 million lives worldwide. There is a need for the identification of therapeutic targets to treat COVID-19. SARS-CoV-2 spike is a target of interest for the development of therapeutic targets. We developed a robust SARS-CoV-2 S spike expression and purification protocol from insect cells and studied four recombinant SARS-CoV-2 spike protein constructs based on the original SARS-CoV-2 sequence using a baculovirus expression system: a spike protein receptor-binding domain that includes the SD1 domain (RBD) coupled to a fluorescent tag (S-RBD-eGFP), spike ectodomain coupled to a fluorescent tag (S-Ecto-eGFP), spike ectodomain with six proline mutations and a foldon domain (S-Ecto-HexaPro(+F)), and spike ectodomain with six proline mutations without the foldon domain (S-Ecto-HexaPro(-F)). We tested the yield of purified protein expressed from the insect cell lines Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tni) and compared it to previous research using mammalian cell lines to determine changes in protein yield. We demonstrated quick and inexpensive production of functional glycosylated spike protein of high purity capable of recognizing and binding to the angiotensin converting enzyme 2 (ACE2) receptor. To further confirm functionality, we demonstrate binding of eGFP fused construct of the spike ectodomain (S-Ecto-eGFP) to surface ACE2 receptors on lung epithelial cells by flow cytometry analysis and show that it can be decreased by means of receptor manipulation (blockade or downregulation)

Truncated O-Glycan-Bearing MUC16 Enhances Pancreatic Cancer Cells Aggressiveness via α4β1 Integrin Complexes and FAK Signaling

Elevated levels of Mucin-16 (MUC16) in conjunction with a high expression of truncated O-glycans is implicated in playing crucial roles in the malignancy of pancreatic ductal adenocarcinoma (PDAC). However, the mechanisms by which such aberrant glycoforms present on MUC16 itself promote an increased disease burden in PDAC are yet to be elucidated. This study demonstrates that the CRISPR/Cas9-mediated genetic deletion of MUC16 in PDAC cells decreases tumor cell migration. We found that MUC16 enhances tumor malignancy by activating the integrin-linked kinase and focal adhesion kinase (ILK/FAK)-signaling axis. These findings are especially noteworthy in truncated O-glycan (Tn and STn antigen)-expressing PDAC cells. Activation of these oncogenic-signaling pathways resulted in part from interactions between MUC16 and integrin complexes (α4β1), which showed a stronger association with aberrant glycoforms of MUC16. Using a monoclonal antibody to functionally hinder MUC16 significantly reduced the migratory cascades in our model. Together, these findings suggest that truncated O-glycan containing MUC16 exacerbates malignancy in PDAC by activating FAK signaling through specific interactions with α4 and β1 integrin complexes on cancer cell membranes. Targeting these aberrant glycoforms of MUC16 can aid in the development of a novel platform to study and treat metastatic pancreatic cancer