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... 3D Demonstration of Strabismus from Peripheral nerve neuropathies Syed Mohammed Qadri, OMS II MU-COM; David L. Dufeau, PhD MUCOM sqadri679@marian.edu; ddufeau@marian.edu ABSTRACT Strabismus squint eyes is a common disorder presenting in about 2% of children born each year. While there are several causes to this presentation, we demonstrated presentation caused by Peripheral nerve neuropathies, especially the ones originating from Oculomotor (CN III) and Abducens Nerve (CN VI). The goal of this project is to create explicit 3D videos and models to be used for student and patient education. Additionally, the pathology and visual defect manifestations of the disorder were also discussed. INTRODUCTION DISCUSSION RESULTS Figure 3: A 3D image of Hypotropia is shown using model eye and eyeball in Coronal view Using advanced 3D rendering softwares, our goal is to explain extraocular muscle disorders in a manner that is easy for everyone to understand. Strabismus often times presents with children, and understanding the disorder is key for parents that may lead to better treatment outcomes through early intervention. Figure 4: Figure 3 is rotated into axial view to show the affected muscle and nerve in cases of Hypotropia Figure 5: Depicted is an example of how Hypotropia can have an affect on a patients vision: Diplopia (double vision) SCAN ME METHODS The goal of this project was to make a learning tool for medical students and parents alike. Being a disorder than presents at a very young age, strabismus can not only have impacts on visual health, but also social health. Strabismus treatment has been proven to have better outcomes with early intervention, therefore, educating parents on seeking treatment early in the life of an infant is key to having the best outcomes that can avoid issues like double vision. Additionally, squint eyes can also have a negative impact on a childs day to day life. Research has shown that kids are less likely to play and make friends with weirdlooking eyes. This negative attitude of peers among young age can negatively impact overall growth of a child. CONCLUSION Accessible 3D models for medical education has opened new ways of learning gross anatomy. With daily technological advancements, 3D visualization is even used by surgeons before performing a major surgery. Through this project, we hope to add to 3D learning by providing accessible and comprehensive teaching tool for medical students and parents. FUTURE PLANS Figure 1: MRI scan used to model 3D render Figure 2: 3D model of head, eye, and eye muscles made using Figure 1 MRI data obtained from TCIA and analyzed in Horos DICOM viewer was exported to Amira for 3D modeling. After converting 2D image data into 3D and manually designing some anatomy by referring to Thieme Atlas of Anatomy, Meshlab was used to further enhance the visuals on the renders. Amira was used again to create animations and compose videos, which were later exported to Camtasia for further stitching and editing in order to demonstrate the different kinds of Strabismus presentations: Hypo-, Hyper-, Exo-, and Esotropias. Anterior Posterior Figure 6: Scan the QR Code to go to a YouTube video of different strabismus presentations The goal from here on is to make 3D models of other structures in the human body, especially ones that are rather difficult to visualize: Brain stem, Diencephalon, Prostate, etc. LINKS Figure 7: This is a 3D model pictured in sagittal view with extraocular muscles labeled https://www.youtube.com/watch?v=weKu1RPLHN0 CONCLUSION This research project was made possible by Marian University College of Osteopathic Medicine 3D Visualization Lab run by Dr. David Dufeau.  ...

... The Role of CaMKK2 in Whole-Body Metabolism and Bone Remodeling Sheel Patel3 , Justin Williams1,2, and Uma Sankar 1,2 of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, 2Indiana Center for Musculoskeletal Health, 3Indiana University School of Medicine, Indianapolis, IN 46202, USA. Abstract Obesity and metabolic dysfunction negatively impacts many tissues, including the skeleton. Calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2) is a serine/threonine protein kinase and key regulator of whole-body energy homeostasis. Previous studies have shown that male Camkk2-/- mice are protected from high-fat diet induced obesity, glucose intolerance and insulin resistance. CaMKK2 regulates hypothalamic control of appetite, hepatic gluconeogenesis, and insulin production by pancreatic cells. Deletion of CaMKK2 has also been shown to enhance bone mass and strength in mice. The skeleton is recognized as a highly dynamic and metabolic tissue capable of regulating systemic glucose metabolism. The role of CaMKK2 in bone-mediate regulation of whole-body metabolism is unknown. Figure 1. Conditional deletion of CaMKK2 from osteoblasts and osteocytes protects from diet-induced obesity. 1A. 1B. 2A. 80 INDIANA UNIVERSITY SCHOOL OF MEDICINE Hypothesis We hypothesize that bone-targeted deletion of CaMKK2 from osteoblasts and osteocytes will concurrently improve glucose metabolism under normal and high fat diet and will simultaneously promote bone accrual. Materials & Methods Animal studies were approved by the Indiana University School of Medicine (IUSM) Institutional Animal Care and Use Committee (IACUC). To generate DMP1-Cre: Camkk2fl/fl (Camkk2OCY) and Dmp1-8kb-Cre+: Camkk2+/+ (Control) mice, we used the Cre-LoxP recombination system and crossed Camkk2fl/fl and dentin matrix protein 1 (Dmp1-8kb)-cre transgenic mice. Eight-week-old Camkk2+/+, , Camkk2-/-, ControlOCY, and Camkk2OCY mice (n=12/group) were randomly assigned to high-fat (60% kcal from fat), or nutrient matched control (10% kcal from fat) diets for a duration of 16 weeks. Diets contained equal amounts of protein, sucrose, vitamins, and minerals. Bbone mineral density (BMD) and body composition were measured using dual-energy Xray absorptiometry (DEXA) beginning two weeks prior to the initiation of diets and monitored biweekly for an additional 16 weeks. Metabolic phenotyping including glucose tolerance test (GTT) and insulin tolerance test (ITT) was performed on male mice (n=12/group) prior to beginning diets and tested again after 16 weeks on diets. Following 16 weeks of control or high-fat diet (HFD) mice were euthanized and long bones were excised for analysis. Right femurs (n=6-7/group) were fixed in 4% paraformaldehyde (PFA) for 48 hours and transferred to 70% ethanol. The midshaft and distal femur microarchitecture were analyzed using microcomputed tomography (CT). Statistical comparisons between groups were made using a two-way ANOVA. All values are presented as means standard deviation. p-value < 0.05 was considered significant. Figure 2. Global deletion of CaMKK2 reduces cortical bone porosity. Porosity (%) 1Department 60 40 20 0 Control Diet High Fat Diet Camkk2+/+ Camkk2-/- (1A.) Changes in body weight measured biweekly for ControlOCY and Camkk2OCY on control and high fat diet. (1B.) Changes in body weight measured biweekly for WT and Camkk2-/- on control and high fat diet. Values displayed represent mean standard deviation (n = 7/group; **** p<0.0001, and *p<0.01). (2A.) Changes in Closed Porosity for WT and Camkk2OCY following 16 weeks of either control or high-fat diet. Values displayed represent mean standard deviation (n = 7/group; ** p<0.01, and *p<0.01). Results Camkk2OCY showed significantly lower weight compared to Control OCY in the high-fat diet group starting at 8 weeks (Fig 1A). Camkk2OCY showed significantly lower weight compared to Control OCY in the control diet group at weeks 10 and 12 only (Fig 1A). The only timepoint where Camkk2-/- had significant lower weight compared to WT was at week 6 in the high-fat diet group. There were no changes in the control diet groups. (Fig 1B). Closed porosity was significantly decreased in Camkk2-/- compared to WT in control and high-fat diet groups (2A). Discussion Conditional deletion of CaMKK2 from osteoblasts and osteocytes protected male mice from diet induced obesity, while global ablation of CaMKK2 did not. These results were somewhat unexpected, as previous studies have shown Camkk2-/- mice have are protected from diet-induced obesity; however, it is important to note fat content and duration of diets used in the study is different. Our results indicate that CaMKK2 and the skeleton play a key role whole-body metabolism. Closed porosity was significantly decreased in Camkk2-/- mice suggesting that CaMKK2 also plays a protective role in bone in the setting of diet changes. This project is still ongoing, and more data and findings are forthcoming. Department of Anatomy, Cell Biology and Physiology  ...

... Role of CaMKK2 in Mechanical Stimulation of Bone Sheel Patel3, Jennifer Shutter1,2, Adam Warrick3, Margaret Bello3, Justin Williams1,2, Alexander Robling1,2, Teresita Bellido1,2, William R. Thompson1,2, and Uma Sankar 1,2 1Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, 2Indiana Center for Musculoskeletal Health, 3Indiana University School of Medicine, Indianapolis, IN 46202, USA. Introduction There is a need for anabolic therapies to treat degenerative bone diseases. Mechanical loading stimulates anabolic and inhibits catabolic pathways of bone leading to increased bone formation. Osteocytes are the bone cell type responsible for coordinating the mechano-stimulatory pathways in bone. Calcium/ calmodulindependent protein kinase kinase 2 (CaMKK2) is a serine/threonine protein kinase with roles in bone remodeling. CaMKK2 global deletion results in lower osteoclast differentiation and higher osteoblast formation and activity. Hypothesis We hypothesize that global and osteocyte specific loss of CaMKK2 in female mice will result in higher bone formation under mechanical loading compared to non-loaded limbs. A B A INDIANA UNIVERSITY SCHOOL OF MEDICINE ns ns C D ns ns Bone formation rate/ bone surface (BFR/BS) between loaded Dmp1Cre: Camkk2-/- and Cre+-only mice and WT loaded and CaMKK2 loaded mice did not show any significant changes. Quantification for loaded tibia. N=6; mean SD; p-values: **** p<0.0001, and ***p<0.001. Materials & Methods Animal studies were approved by the Indiana University School of Medicine (IUSM) Institutional Animal Care and Use committee (IACUC). Conditional deletion of Camkk2 in osteocytes was achieved by crossing Camkk2flox/flox mice with dentin matrix protein1 (Dmp1 -8kb, osteocyte-specific) -Cre transgenic mice, to generate Dmp1-Cre:Camkk2-/- (osteocyte-specific) and Dmp1-Cre+ (Cre+ only) control mice. The right tibiae of 18-week-old female WT, Camkk2-/-, Dmp1-Cre+, and Dmp1-Cre: Camkk2-/- mice were loaded at 2 Hz for 220 cycles while anesthetized and peak forces were predetermined for each genotype. The left tibiae served as non-loaded internal controls. Mice were loaded on days 1, 3, 5, 8 and 10 of the experiment using an electro actuator (Bose ElectroForce 3200; EnduraTEC, Minnetonka, MN, USA). Calcein and alizarin red were administered intraperitoneally on days 9 and 16 respectively to measure bone formation using dynamic histology. Mice were sacrificed on day 19. Left and right tibiae were collected and fixed in 4% paraformaldehyde for 48 h and stored at 4C in 70% ethanol. Micro-computed tomography (CT) was performed on both loaded and non-loaded tibiae to analyze bone microstructure in the proximal tibia and cortical midshaft. (N=10/genotype). Undecalcified tibiae were embedded in polymethyl methacrylate and cross-sections were taken of the tibial midshaft for dynamic histology (N=5/genotype). Statistical comparisons between groups were made using Welchs t-test . All values are presented as means standard deviation. p-value <0.05 was considered significant. Figure 2. CaMKK2 effect on bone formation rate in mechanically loaded and non-loaded female mice Figure 1. Impact of global or osteocyte-specific loss of CaMMK2 in mechanically loaded female mice E F ns ns (1A) Camkk-/- and Dmp1-Cre: Camkk2-/- mice showed significantly higher levels of (BV/TV %), (1B) trabecular number , and (1C) cortical midshaft thickness when compared to WT and Cre+-only control mice. Discussion Global and osteocyte-specific deletion of CaMKK2 in female mice have higher %BV/TV, higher Tb. N, higher Ct. Th, compared to WT and Cre+only control mice. Importantly, BFR/BS did not show any changes between all genotypes. These findings suggests while CaMMK2 does impact bone microstructure in loaded mice, it plays no role in bone mass accrual in mechanically stimulated mice. Further understanding of the role that CaMKK2 plays in the process of bone remodeling may allow the development of novel therapies to target various degenerative disease states. *This project is still on-going and more data has yet to be analyzed. In addition, the role of CaMMK2 in mechanically stimulated males is concurrently being investigated by our lab.* (1D) Trabecular thickness and (1E) mineral apposition rate did not show any significant changes between all genotypes. (1F) There was no significant change in mineral apposition among all genotypes. Quantification for loaded tibia. N=10; mean SD; p-values: **** p<0.0001, and **p<0.01. Acknowledgements This work was funded by __________ to US, and Moenkhaus fellowship (IUSM) Department of Anatomy, Cell Biology and Physiology  ...

... Temporal Study of Renal Volume Losses in Patients with Robotic Partial Nephrectomies 1Rushi Patel, MS, 1Chandru P. Sundaram, MD, MS, 2Tsunenori Kondo, MD, PhD, 1Clinton D Bahler, MD, MS 1Indiana University School of Medicine, Department of Urology, Indianapolis, IN 2Tokyo Womens Medical University, Department of Urology, Tokyo, Japan Abstract Materials & Methods Purpose: Partial nephrectomies by their nature are associated with renal volume loss. Our goal from this study is to examine renal volume loss over time post partial nephrectomy. Material and Methods: Fifty patients were followed for 1-year post partial robotic nephrectomy with two-layer renorrhaphy and the sliding clip technique. This was done with an initial preoperative computed tomography (CT) scan to assess renal mass and location. Post partial nephrectomy patients were imaged at time points 3-days, 6-months, and 12-months. Results: Patient demographics were 82% male with a median (IQR) age of 57 (45-67) and all were of Japanese descent. The medians (IQR) for warm ischemia time: 18 minutes (14-22), total operative time: 181.5 minutes (169.3-218.5), and estimated blood loss: 20 mL (10-50). The tumor characteristics had a median (IQR) diameter of 2.8 cm (2.5-3.4) with a RENAL score of 7 (6-8). The renal CT volumes showed median (IQR) volume losses at 3-days: -1% (-7.1, 1.8), 6-months: 15.3% (-20.6, -11.2), and 12-months: -16.3% (-19.0, -12.8). Significance was seen at the 3-days to 6-months comparison for volume loss (p<0.0001). Mean (SD) eGFR losses were as follows: at discharge 0.5% (12.9), 1-month -6.4% (11.8), 6-months -4.6% (9.8), and 12-months -3.6% (11.9). Statistical analysis showed significance for GFR loss at the comparison between discharge to 1month and 6-months (p=0.01, p=0.04). Conclusion: The initial volume loss seen post-surgery from resected healthy tissue was not significant and only became relevant at longer time points suggesting that loss could be from atrophy. Volume loss over time supports the hypothesis that suture renorrhaphy is a primary cause of volume loss when warm ischemia time is <25minutes. Background Small renal masses are increasingly being treated with partial nephrectomy as availability of robotic assistance increases. Emphasis is placed on sparing renal function which includes minimizing resected healthy kidney and ischemic injury from suture renorrhaphy and hilar clamping. The degree to which resection of healthy parenchyma or renorrhaphy contributes to renal volume loss not well characterized. Our hypothesis is that postoperative day 3-4 CT scans demonstrate resection loss and delayed scans at 4-6months demonstrate ischemia/renorrhaphy injury and that by comparing the two we can better understand volume and functional loss after partial nephrectomy. A total of 50 patients undergoing robotic-assisted partial nephrectomy by a single surgeon between 4/2013 and 3/2015 had available CT scans for volume calculation preoperatively, postoperative day 3, 6 months, and 12 months. Those with tumors <2cm, multiple tumors, or requiring postoperative embolization were excluded. Intraoperative US was used in all the cases and vascular bull dogs were used for clamping. A sliding clip cortical renorrhaphy was used in all cases. There were no transfusions or intraoperative complications. The post-operative day 3 or 4 scan was obtained prospectively to assess for asymptomatic pseudoaneurysms. A base-layer and cortical renorrhaphy were used. Philips Intellispace Portal was used to construct three-dimensional models of the operated kidneys using slice-by-slice semi-automatic segmentation. The vol. of the tumor was subtracted from the preoperative vol. GraphPad Prism was used to perform descriptive statistics and the Mann-Whitney U-test to compare volume losses. Table 1. Demographics n Age, years Male, % Japanese Descent, % Right side, % BMI, kg/m2 Hemoglobin, g/dL Tumor diameter, cm Nephrometry, RENAL Low (4-6), % Intermediate (7-9), % High (10-12), % Positive surgical margins, % Pathology Renal cell carcinoma Clear Cell Papillary Chromophobe Mucinous, tubular, and spindle cell Tumor Stage carcinoma pT1a (%) pT1b (%) Nodal Metastasis N0 Distant Metastasis M0 Fuhrman Grade One Two Three Four Median (IQR) 50 57 (45-67) 82% 100% 54% 24.7 (23-26) 14.6 (13.98-15.40) 2.8 (2.5-3.4) 7 (6-8) 19 (38%) 27 (54%) 4 (8%) 0% n (%) 50 (100) 44 (88) 3 (6) 2 (4) 1 (2) Table 2. Surgical Intervention Results for Robotic Partial Nephrectomies Median (IQR) Total operating time (from incision to finished closing), min. 181.5 (169.3-218.5) EBL, mL 20 (10-50) 2 eGFR pre-op, mL/min/1.72m 65.8 (56.7-75.1) Warm ischemia time, min. 18 (14-22) Urine leaks, % 2% Drains placed, % 52% Table 3. Volumetric and Functional Losses cm3 Relative Volume loss at 3-4 days, Relative Volume loss at 6 months, cm3 Relative Volume loss at 12 months, cm3* Percent Volume loss at 3-4 days, % Percent Volume loss at 6 months, % Percent Volume loss at 12 months, %* Relative GFR loss at discharge, mL/min/1.72m2 Relative GFR loss at 1 month, mL/min/1.72m2 Relative GFR loss at 6 months, mL/min/1.72m2 Relative GFR loss at 12 months, mL/min/1.72m2 Percent GFR loss at discharge, % Percent GFR loss at 1 month, % Percent GFR loss at 6 months, % Percent GFR loss at 12 months, % *limited to 20 with available CT scan Median (IQR) -1.9 (-11.2, 2.8) -23.5 (-34.3, -17) -23.3 (-30.6, 19.3) -1.0 (-7.1, 1.8) -15.3 (-20.6, 11.2) -16.3 (-19.0, 12.8) Mean (STD) -0.5 (11.8) -6.4 (11.7) -4.8 (9.3) -3.7 (10.6) 0.5 (12.9) -6.4 (11.8) -4.5 (9.8) -3.6 (11.9) Figure 2: (A) Volumetric loss post partial nephrectomy showing significant volume loss at 6months and volume loss stabilization at 12-months time point. (B) GFR loss post partial nephrectomy showing significant loss after 6-months and insignificant changes in 6-to-12month comparison. Discussion Conclusion The majority of volume loss after partial nephrectomy was not detected in the immediate postoperative period but was seen at 6months. This supports the hypothesis that volume and functional loss after partial nephrectomy are not due to resected healthy kidney alone but also ischemia and reconstruction injury. Further studies are warranted evaluating renal function loss after partial nephrectomy looking at reconstructive injury in addition to resection related loss. Limitations A single partial nephrectomy technique is evaluated in a small series. Postoperative renal hyperemia or edema could have artificially elevated the 3-4day volume results although this appears to be minimal. The resected healthy margin was not available for evaluation although the attempted margin was ~5mm. References 45 (90) 5 (10) 50 (100) 50 (100) 6 (12) 40 (80) 3 (6) 1 (2) Results Figure 1: CT imaging of the abdomen. The selected sections indicate the renal parenchyma during contrast infusion at 3 time points with estimated renal volumes. (A) Pre-Operative. (B) Immediately post-surgery. (C) 6 months post-surgery. (D) 3D reconstruction of kidney pre-surgery used for volume estimation. [1] Sun M, Abdollah F, Bianchi M, et al. Treatment management of small renal masses in the 21st century: A paradigm shift. Ann Surg Oncol 2012; 19: 23802387. [2] Dagenais J, Maurice MJ, Mouracade P, et al. Excisional precision matters: understanding the influence of excisional volume loss on renal function after partial nephrectomy. Eur Urol 2017; 72: 168170. [3] Bahler CD, Dube HT, Flynn KJ, et al. Feasibility of omitting cortical renorrhaphy during robotic partial nephrectomy: a matched analysis. J Endourol 2015; 29: 548555. [4] Morita S, Tajima T, Yamazaki H, et al. Early postoperative screening by contrast-enhanced ct and prophylactic embolization of detected pseudoaneurysms prevents delayed hemorrhage after partial nephrectomy. J Vasc Interv Radiol 2015; 26: 950957. [5] Dong W, Wu J, Suk-Ouichai C, et al. Devascularized parenchymal mass associated with partial nephrectomy: predictive factors and impact on functional recovery. J Urol 2017; 198: 787794. [6] Bahler CD, Cary KC, Garg S, et al. Differentiating reconstructive techniques in partial nephrectomy: a propensity score analysis. Can J Urol 2015; 22:7788-7796. [7] Takagi T, Kondo T, Tachibana H, et al. Comparison of surgical outcomes between resection and enucleation in robot-assisted laparoscopic partial nephrectomy for renal tumors according to the surface-intermediatebase margin score: a propensity score-matched study. J Endourol 2017; 31:756-761. [8] Baumert H, Ballaro A, Shah N, et al. Reducing warm ischaemia time during laparoscopic partial nephrectomy: a prospective comparison of two renal closure techniques. Eur Urol. 2007; 52:1164-1169. [9] Kondo T, Takagi T, Morita S, et al. Early unclamping might reduce the risk of renal artery pseudoaneurysm after robot-assisted laparoscopic partial nephrectomy. Int J Urol. 2015; 22:1096-1102. [10] Azhar RA, Bochner B, Catto J, et al. Enhanced recovery after urological surgery: a contemporary systematic review of outcomes, key elements, and research needs. Eur Urol. 2016; 70:176-187. [11] Cerantola Y, Valerio M, Persson B, et al. Guidelines for perioperative care after radical cystectomy for bladder cancer: enhanced recovery after surgery (ERAS((R))) society recommendations. Clin Nutr. 2013; 32:879-887. [12] Matsuo S, Imai E, Horio M, et al. Revised equations for estimated GFR from serum creatinine in Japan. Am J Kidney Dis. 2009; 53: 982-992 [13] Simmons MN, Lieser GC, Fergany AF, et al. Association between warm ischemia time and renal parenchymal atrophy after partial nephrectomy. J Urol 2013; 189: 16381642. [14] Takagi T, Mir MC, Sharma N, et al. Compensatory hypertrophy after partial and radical nephrectomy in adults. J Urol 2014; 192: 1612-1618. [15] Simmons M, Fergany A and Campbell S. 542 Baseline gfr and percent functional volume preservation are the primary determinants of long-term gfr after partial nephrectomy. J Urol 2011; 185: e220. [16] Tanaka N, Fujimoto K, Tani M, et al. Prediction of postoperative renal function by preoperative serum creatinine level and three-dimensional diagnostic image reconstruction in patients with renal cell carcinoma. Urology 2004; 64: 904908. [17] Shekarriz B, Shah G and Upadhyay J. Impact of temporary hilar clamping during laparoscopic partial nephrectomy on postoperative renal function: A prospective study. J Urol 2004; 172: 5457. [18] Bahler CD and Sundaram CP. Effect of renal reconstruction on renal function after partial nephrectomy. J Endourol 2016; 30: S37S41. [19] Sarma A, Heilbrun ME, Conner KE, et al. Radiation and chest ct scan examinations: what do we know? Chest 2012; 142: 750760.  ...

... Utility of Classroom Subject Performance Outcomes in Preparation for the Comprehensive Osteopathic Medical Licensing Examination (COMLEX) Rachna Chaudhari & Sarah Zahl, PhD Marian University College of Osteopathic Medicine, Indianapolis, Indiana Introduction Results cont. Correlations exist between undergraduate GPA, MCAT, and COMLEX-USA Level 1 scores as well as between course grades and overall COMLEX score (Wong et al. 2009 & Sefcik et al. 2003). Longitudinal reports on ProgressIQ contain aggregate data over two years of didactic work in the first two years of medical school. Classroom subjects divided into systems report an average score over two years. NBOME provides COMLEX-USA Level 1 score reports for each examiner which include discipline sub-scores for each system. Methods Class of 2021 and 2022 COMLEX data was deidentified and used in this study. Any longitudinal report category with <30 items was removed. Correlations and p values were calculated using Microsoft Excel Data Analysis Toolkit. Discipline Anatomy* Anat/Embryology* (Anat/Histology) Biochemistry* Microbiology* Microbiology/Imm unology* Osteopathic Principles & Practice* Pathology* Pharmacology* Physiology* (Behavioral Science)^ (Public Health) Biosystems Community Wellness* Human Development; Reproductive; Sexuality* Endocrine* Nervous System; Mental Health* Musculoskeletal* Genitourinary* Gastrointestinal; Nutrition* Circulatory; Hematology* Respiratory* Integumentary* Table 2. C2022 (n=163) Correlation Results FOMCD & Blooms Medical Knowledge* Osteopathic Manipulative Medicine/ Osteopathic Principles and Practice* Patient Care Procedural* (Professionalism) Blooms 1/2* Blooms 3/4* Blooms 5/6* Discussion and Future Direction Even though discipline sub-scores are based on a relatively lower number of items and scores may be less reliable, a statistically significant moderatestrong correlation exists for almost every category. Behavioral Science exam questions should be investigated and compared with NBOMEs blueprint. Survey to C2021 and C2022, 30% response rate: Inquired about study habits When asked if longitudinal reports are used to tailor study plans: Yes Results r 0.50 indicates a positive correlation (); not correlated. *; moderately-strongly correlated. Red; correlated to overall COMLEX score not subscore. ^; not statistically significant. Discipline Anatomy* Embryology* Histology* Biochemistry* Microbiology* Immunology* Osteopathic Principles and Practice* Pathology* Pharmacology* Physiology* (Behavioral Science)^ Diagnosis and Management* (Medical Ethics) Biosystems (Asymptomatic) Cardiovascular* Endocrine* Foundational* Gastrointestinal* Genitourinary* Head and Neck* Hematology Oncology* Integumentary* Musculoskeletal* Nervous System* Reproductive System* Respiratory System* Table 1. C2021 (n=155) Correlation Results 62% N/a did not know about reports By identifying weaker areas in the curriculum and targeting those areas, it can potentially improve sub-score outcomes which impacts overall performance. Since COMLEX-USA is becoming Pass/Fail in May 2022, making sure students have a strong understanding in each area of the curriculum will be even more important to ensure passing the examination. Future studies should analyze student data at other osteopathic medical schools to see if similar correlations exist at institutions utilizing different curriculums. FOMCD & Blooms Medical Knowledge* Osteopathic Manipulative Medicine/ Osteopathic Principles and Practice* Patient Care Procedural* (Professionalism) Blooms 1/2* Blooms 3/4* Blooms 5/6* No 24% 14% References 1. Sefcik, DJ et al. (2003). Characteristics of the Courses That Best Predict COMLEX-USA Level 1 Performance. JAOA, 103(10), 491-494. 2. Wong, SK et al. (2009). Student Performance on Levels 1 and 2-CE of COMLEX-USA: Do Elective Upper-Level Undergraduate Science Courses Matter? JAOA, 109(11), 592-598. Acknowledgements Thank you ProgressIQ, Wendy Labuzan, C2021, and C2022.  ...

... Development of a Registry to Identify Patent Ductus Arteriosus Treatment Patterns in Preterm Infants at Peyton Manning Childrens Hospital Rachel Segar, MS, Zenaida Tauscher, MD and Markus Tauscher, MD ABSTRACT Aim: To address this knowledge gap, a retrospective, observational cohort study was designed to evaluate the clinical circumstances leading to an echocardiographic evaluation for a PDA, to collect information on how a PDA is described echocardiographically, the management of the PDA, and the impact on morbidity and mortality of extremely preterm infants. Methods: Data will compare morbidity and mortality for 1) infants, who were never evaluated for a PDA by ECHO, 2) infants, who were evaluated for a PDA by ECHO, but had no PDA, 3) infants, who were diagnosed with a PDA by ECHO and received treatment, and 4) infants, who were diagnosed with a PDA by ECHO but did not receive treatment. Clinical circumstance leading to Echocardiographic evaluation Description of Echo findings Treatment for PDA Indomethacin Ibuprofen Acetominophen Ligation Catheterization Maternal Data Demographic data Neonatal morbidities Severe Intraventricular hemorrhage (IVH grade III and IV Periventricular leukomalacia Bronchopulmon ary Dysplasia Retinopathy of Prematurity requiring treatment Necrotizing Enterocolitis Spontaneous intestinal perforation Sepsis Pneumonia Cardiovascular data Problem: In the neonatal intensive care unit (NICU) at Peyton Manning Childrens Hospital (PMCH), there are no standardized criteria for screening for a PDA, echocardiographic description of a PDA or treatment of a PDA. Ordering confirmatory echocardiography (ECHO) for a suspected PDA and treatment is dependent on the attending neonatologist. Data collection Neonatal Data Background: A patent ductus arteriosus (PDA) is a common diagnosis among extremely preterm infants. Unfortunately, this left to right shunt may result in significant complications including pulmonary edema, pulmonary hemorrhage, bronchopulmonary dysplasia, as well as intraventricular hemorrhage, and necrotizing enterocolitis. Notably, there is no consensus on treatment for a PDA. As such, the diagnosis and treatment of a PDA have varied considerably between practitioners and institutions. Demographic Maternal morbidity Medications Antenatal Steroid therapy Antenatal magnesium Duration of rupture of membranes Mode of delivery RESEARCH DESIGN AND METHODS The proposed investigation is a retrospective, observational cohort study. The investigators will extract data from the healthcare software Neodata (Isoprime Corporation, Lisle, IL) and the electronic medical record Allscripts Sunrise (Allscripts Healthcare, LLC, Chicago, IL). Subgroups of interest St. Vincent Peyton Manning Children's Hospital Infants who were never evaluated for a PDA by ECHO Infants who were evaluated for a PDA by ECHO, but had no PDA Infants who were diagnosed with a PDA by ECHO and received treatment Infants who were diagnosed with a PDA by ECHO but did not receive treatment Anticipated results Through this work, we aim to gain an understanding of treatment and management patterns of PDAs at Peyton Manning Childrens Hospital. Such work may help develop a standardized protocol for consistent PDA treatment in the future. References: Phillips, J. B., III. (2021, May). Patent ductus arteriosus in preterm infants: Pathophysiology, clinical manifestations, and diagnosis. Retrieved June 09, 2021, from https://www.uptodate.com/contents/patent-ductus-arteriosus-inpreterm-infants-pathophysiology-clinical-manifestations-anddiagnosis?search=pda+neonate&topicRef=15814&source=see_l ink  ...

... The Role of GM-CSF in the Neuropathogenesis of Simian Immunodeficiency Virus (SIV) Rachael Meckley, BMS Thesis Student Advisor: Dr. Elizabeth Delery, PhD Introduction Results Why do neurocognitive disorders persist in people living with HIV while taking combination antiretroviral therapy (cART)? The brain is a known reservoir for HIV. HIV can act on myeloid cell populations like macrophages. Granulocyte Macrophage Colony Stimulating Factor (GM-CSF) is a hematopoietic growth factor and type I soluble cytokine that has been demonstrated to have a proinflammatory role over macrophages (and other myeloid cells) and can induce proliferation and migration across the blood brain barrier (Vogel 2015). GM-CSF has also been found to be present and is involved in the inflammation process of Simian Immunodeficiency Virus (SIV). Along with GM-CSF, the expression of GM-CSF receptors has been shown to be increased in SIV. Fig. 5: Quantification of CD163 in animal 17-0780 (left) and animal 150050 (right). Fig. 1: Percentage of GM-CSF positive cells per the number of nuclei. Fig. 2: Percentage of CSF2RB positive cells per the number of nuclei. Fig 6: Percent of cells positive for CD163+ per the number of nuclei present in an SIV positive (17-0480) and an SIV negative (150050 animal. Hypothesis: In SIV infected regions of the midbrain, GM-CSF expression will be increased, which would contribute to the HIV neuropathogenesis by attracting inflammatory cells such as macrophages. Methods Conclusion Sections of midbrain were obtained from ten Rhesus Macaque animals in formalin fixed, paraffin embedded tissue slides. Immunohistochemistry (IHC) was performed on five SIV positive slides and five SIV negative slides of Rhesus Macaque brain tissue. Primary Antibody Used Specificity GMCSF (1:100) Granulocyte Macrophage Colony Stimulating Factor CD163 Macrophage Marker Phalloidin Structural Protein DAPI Nuclei CSF2RB GM-CSF Receptors Images were taken on 4x, 10x, 40x and 100x DAPI was counted via a Macro in FIJI GMCSF and CSF3RB were manually counted in FIJI Percent of the number of positive GMCSF cells were counted per the number of positive nuclei. Calculations from these analyzed images were performed in excel. Statistic graphs were made in Graph Pad Prism Material for counting GM-CSF and CSF2RB was included if there was a DAPI nuclei present with the red or far-red cytokine/receptor (Fig. 1 and Fig. 2) Material for counting was included if there was both a blue nucleus (DAPI) and far-red macrophage (CD163) present in the same location (Fig. 5). Increased expression of GM-CSF and receptors in SIV positive animals. GM-CSF has a proinflammatory role over myeloid cells including macrophages. Brain is a reservoir for HIV. HIV can influence macrophages and can play a role in the viral persistence of HIV. Increased GM-CSF expression can lead to increased expression of macrophages (CD163)? Infected macrophages could spread HIV by using the ability to invade most body tissues. Increased amounts of macrophages can cause neuroinflammation and contribute to the pathophysiology of HIV in the brain. To help patients living with HIV and suffering from neurocognitive impairments, this research could present a potential treatment created by combining GM-CSF antibodies with antiretroviral therapies in the future. Fig. 3: Representative images of SIV positive animal 14-0681. GM-CSF is represented by red (top left), CSF2RB is represented by far-red (top right), and DAPI are represented in blue (bottom left. The last image is the overlay (bottom right). Fig. 4: Representative images of SIV positive animal 15-0432. GM-CSF is represented by red (top left), CSF2RB is represented by far-red (top right), and DAPI are represented in blue (bottom left. The last image is the overlay (bottom right). Works Cited Vogel, Daphne Y. S. Kooji, Gijis. Heijnen, Priscelilla D. A. M. Breur, Marjolein. Peferoen, Laura A. N. Van der Valk, Paul. De Vries, Helga E. Amor, Sandra. Dijkstra, Christine D. 2015. GM-CSF promotes migration of human monocytes across the blood brain barrier. European Journal of Immunology.  ...

... Surgical Therapy to Atlantoaxial Instability Paul Cooke, OMS II MU-COM and David Dufeau, PhD MU-COM Pcooke508@marian.edu ddufeau@marian.edu; Results Introduction Atlantoaxial instability (AAI) is a term that is used to describe instability at the atlantoaxial joint of the cervical spine. This is often from alar ligament instability causing the dens of the axis to be prone to move posterior or sublux. The inferior portion of the medulla oblongata and the superior portion of the spinal cord can be compressed by the posterior movement of the dens causing lifethreatening effects. In this presentation, a surgical therapy to atlantoaxial instability will be examined for the purpose of educating future medical students. We will specifically be examining AAI with spina-bifida occulta at C1. Methods Using the image databank Horos, a file containing CT images of the skull and cervical spine was obtained. This data set contained a patient with apparent spina-bifida occulata at the atlas (C1) causing spinal instability at the atlantoaxial joint. Occiput Atlas (C1) with apparent SpinaBifida Atlas (C1) Discussion Our intent for this project was to use the 3D imaging and printing technology to better educate future medical students about the spinal corrections available for AAI. Additionally, 3D imaging, and printing technology provides unique clarity to anatomical structures. Finally, these images and 3D prints allow for viewing of the therapy for atlantoaxial instability and subluxation. Atlas (C1) with apparent SpinaBifida Dens of Axis (C2) Axis (C2) Figure 3: Presented above left is the 3D visualization of the cervical spine with apparent spina-bifida at the atlas (C1). Figure 6: Presented below is the 3D print of the spine and occiput. Conclusion Figure 4: Presented above right is the 3D visualization of the hardware used to correct the spinal abnormality. The researchers have found that through using the 3D visualization technology, one can create accurate and intricate images of the atlantoaxial joint of the spine and a corrective surgery for AAI. Both the 3D images and the physical 3D print show great promise in teaching future medical students of both spinabifida of the atlas and AAI causing subluxation at the atlantoaxial joint. Atlas (C1) Axis (C2) Axis (C2) Figure 1: Sagittal view of cervical spine CT Figure 2: Transverse view of the atlas CT Additionally, a CT of the corrective-surgical therapy was also obtained. These CT image sets were then analyzed and 3D rendered using Amira. A 3D print was also obtained using the softwares Meshmixer and Formlabs. A 3D print was completed using Form Labs Form 3 Resin printer. Figure 5: Presented above is the 3D visualization of the corrective hardware installed on the spine. Note: unknown surgical artifact likely used to manage CSF in blue and purple. Figure 7: Presented above a 3D print of the cervical spine and ociput with the corrective hardware installed. Future Developments The researchers hope to further develop 3D visualization software for the purpose of medical student education. They plan to promote the use of 3D visualization as a teaching method for medical students.  ...

... Understanding Pennsylvania Tick Population and Tick-borne Disease Dynamics: A Retrospective Analysis of Archived Databases from 2008-2020 Nishi Natalia, Julia Sendatch, Heidi St. John, Richard Stewart, Marcie L. Lehman, Christina Farris, and Alison Luce-Fedrow Biology Department of Shippensburg University United States Naval Medical Research Center Introduction Ticks have been widely studied for their importance in disease transmission. In Pennsylvania, four different ticks (associated with human diseases) are commonly encountered: (1) Dermacentor variabilis (Rocky Mountain Spotted Fever); (2) Ixodes scapularis (Lyme disease); (3) Amblyomma americanum (Ehrlichiosis); and (4) I. cookei (possible Powassan). In addition to the commonly occurring tick species, approximately 25 species have been identified in the state. I. scapularis, which was once primarily found in Central and Eastern PA, can now be found in all counties in PA. In recent years, PA has had the highest confirmed numbers of Lyme disease cases in the United States. The increasing abundance and prevalence of ticks known to vector human pathogens has prompted public health concerns and further research. Despite the devastating consequences, and potentially fatal outcomes of tick-borne diseases, the local/regional distribution of ticks in Pennsylvania is under-investigated with regards to changes in tick populations and tick-borne disease dynamics. Methods Compiled individual databases from 2008-2020 into one master database Date of collection, location, method of collection, species, sex, life stage, engorgement status, and molecular status Quantitative analysis and mapping (ArcGIS Pro version 2.7.3 (Esri, Redlands, CA)) used to conduct comparative analysis of changing geographical distributions, seasonal distribution, and tick life stage Tick-borne microbes (Rickettsia, Ehrlichia, Anaplasma, and Borrelia) were compared for ticks collected from 2008-2016. Sample Site Density Conclusions Results Total Ticks Collected by Year Tick Species Percentage of Total Ticks Collected 2008 2013 2017 131 ticks 72 ticks 875 ticks I. scapularis 6884/17,507 (39.32%) 2010 2014 2018 D. variabilis 2119/17,507 (12.10%) 2 ticks 138 ticks 521 ticks A. americanum 6868/17,507 (39.23%) 2011 2015 2019 Haemaphysalis spp. 112/17,507 (0.64%) 188 ticks 633 ticks 6512 ticks 2012 2016 2020 1442 ticks 93 ticks 6855 ticks Total Table 2. Major species collected included Ixodes scapularis (39.32%), Amblyomma americanum (39.23%), and Dermacentor variabilis (12.10%). Emergence of two previously rare/unknown ticks in Pennsylvania, A. americanum (2017) and Haemaphysalis (2015), with significant prevalence from 2018-2020. 17,507 ticks Table 1 and Figure 1. A total of 17,507 ticks were collected. Major collecting years included 2012 (n = 1,442), 2019 (n = 6,512), and 2020 (n = 6,855), with concentrations centered around western and central Pennsylvania, but a presence in statewide geographical distribution Pennsylvania 2008-2020 Pathogen qPCR positive ticks Rickettsia 169/793 (21.31%) B. burgdorferi 108/852 (12.68%) E. chaffeensis 59/802 (7.36%) A. phagocyptophilum 15/550 (2.73%) Borrelia spp. 143/377 (37.93%) Table 3. Molecular testing (qPCR) of ticks from 2008-2016 demonstrated Rickettsia (21.31%) and Borrelia burgdorgeri (12.68%) as the most commonly detected tick-borne microbes. The views expressed in this article are those of the author and do not necessarily reflect the official policy or position of the Department of the Navy, Department of Defense, the U.S. Government, nor the Henry M. Jackson Foundation for the Advancement of Medicine, Inc. (HJF). Authors, as employees of the U.S. Government (CMF), conducted the work as part of their official duties. Title 17 U.S.C. 105 provides that 'Copyright protection under this title is not available for any work of the United States Government.' Title 17 U.S.C. 101 defines a U.S. Government work is a work prepared by an employee of the U.S. Government as part of the person's official duties. The study protocol was approved by the Shippensburg University Institutional Animal Care and Use Committee (Shippensburg University IACUC protocol #R02-03-19) in compliance with all applicable federal regulations governing the protection of animals and research. Esri, HERE, NPS, HERE, Garmin, USGS, EPA, NPS Further investigation will examine host association of tick species and the molecular status of ticks that are positive or negative for these pathogens from 2016-2020 Esri, HERE, Garmin, NGA, USGS, NPS I. scapularis and D. variabilis have been present in PA since 2008. A. americanum and Haemaphysalis species are emerging species in the state. Rickettsia and B. burgdorferi are most prevalent among the species of ticks tested. E. chaffeensis and A. phagocytophilum are hypothesized to increase in prevalence. Implications Application in the medical field: provide insight on tickborne illness prevalence in PA Provide pertinent background information for future researchers and epidemiologists Future Research Molecular testing of ticks from 2016-2020 Further expanded surveying of ticks in Pennsylvania REFERENCES 1.Steere, A.C., J. Coburn, and L. Glickstein, The emergence of Lyme disease. J Clin Invest, 2004. 2.USACHPPM. Just the facts...Lyme Disease. 2006 April 2006 [cited 2012. 3.CDC.gov, Notifiable diseases/deaths in selected cities weekly information. MMWR, 2009. 4.Adams, D., et al., Summary of Notifiable Diseases United States, 2011. MMWR, 2013. 5.Gardner, S.L., et al., National surveillance for the human ehrlichioses in the United States, 1997-2001, and proposed methods for evaluation of data quality. Ann N Y Acad Sci, 2003. 6.Paddock, C. and J.E. Childs, Ehrlichia chaffeensis: a prototypical emerging pathogen. Clin Microbiol Rev, 2003. 7.CDC.gov. Ehrlichiosis. 2011 [cited 2012]. 8.Chen, S., et al., Identification of a granulocytotropic Ehrlichia species as the etiologic agent of human disease. J Clin Microbiol, 1994. 9.Hall-Baker, P., et al., Summary of Notifiable Diseases - United States, 2009. MMWR, 2011. 10.Barker TL, et al., Serosurvey of Borrelia burgdorferi infections among U.S. military personnel: A low risk of infection. Am. J. Trop. Med. Hyg., 2001. 11.Courtney JW, et al., Molecular characterization of Anaplasma phagocytophilum and Borrelia burgdorferi in Ixodes scapularis ticks from Pennsylvania. J. Clin. Microbiol., 2003. 12.Steere, A., Lyme Disease. N Engl J Med, 2001. 345(2): p. 115-125. 13.Trevejo, R., et al., Evaluation of a two-test serodiagnostic method for community assessment of Lyme disease in an endemic area. Am. J. Trop. Med. Hyg., 2001. 14.CDC.gov. Lyme Disease. 2012 5 April 2012 [cited 2012]. 15.Magnarelli, L., Global importance of ticks and associated infectious disease agents. Clin Microbiol Newsletter, 2001. 16.Fornadel, C.M., et al., High rates of Rickettsia parkeri infection in Gulf Coast ticks (Amblyomma maculatum) and identification of "Candidatus Rickettsia andeanae" from Fairfax County, Virginia. Vector Borne Zoonotic Dis. 2011. 17.Jiang, J., et al., Development of a quantitative real-time polymerase chain reaction assay specific for Orientia tsutsugamushi. Am. J. Trop. Med. Hyg., 2004. 18.Jiang, J., et al., Molecular detection of Rickettsia amblyommii in Amblyomma americanum parasitizing humans. Vector-Borne and Zoonotic Diseases, 2010. 19.Smith, M.P., et al., Bacterial pathogens in ixodid ticks from a Piedmont County in North Carolina: prevalence of rickettsial organisms. Vector Borne Zoonotic Dis. 2010. 20.Courtney, J.W., et al., Multiplex real-time PCR for detection of anaplasma phagocytophilum and Borrelia burgdorferi. J Clin Microbiol, 2004. 21.Jiang, J., et al., Phylogenetic analysis of a novel molecular isolate of spotted fever group Rickettsiae from northern Peru: Candidatus Rickettsia andeanae. Ann N Y Acad Sci, 2005.  ...

... An Evaluation of Antibiotic Resistance in Bacteria of the White River Waterways: Water Sampling (Poster 1) Nicole Lisek, Kevin Medernach, Azeem Ahmad*, and Samina Akbar* College of Osteopathic Medicine and College of Arts and Sciences, Marian University, 3200 Cold Spring Road, Indianapolis, IN 46222 USA *Co-Investigators ABSTRACT The dramatic increase in the prevalence of antibiotic resistant (AR) genes in numerous bacterial species has been a heavily discussed topic within the scientific community. Antibiotic resistant bacteria pose a serious threat to an individuals health and is an ongoing area of research with a variety of public health implications. Antibiotic resistant genes can lead to the development of super bugs which can become immune to currently used antibiotics. This leads to much more severe infections, difficulties in properly treating those infections, and can result in longlasting health complications. There are several factors responsible for the development of these super bugs, namely over prescription and the spread of medical and agricultural runoff in natural environments. This runoff exposes bacteria to antibiotics and allows them to develop resistance and spread rampantly between non-pathogenic and pathogenic bacteria. Identifying local reservoirs that house these AR bacterial strains has become pertinent. The white river watershed flows through thousands of miles of streams and local drainage areas providing drinking water to residents of central Indiana. The Nina Mason Pulliam Ecolab (NMPE) at Marian University is part of this specific watershed. This Ecolab provides a prime environment in which different bacterial strains can thrive. This provides a crucial medium for the mixing, evolution, and spread of antibiotic resistant genes and bacteria. Gathering and studying water samples from the NMPE will provide insight into the prevalence of multidrug resistance genes. Previous research at MU-COM has found bacteria carrying multiple drug resistance (MDR) in the Nina Mason Pulliam EcoLab (NMPE) watershed. In the summer of 2020, 87.5% of the samples collected exhibited multiple drug resistance. METHODS Isolate # Gram +/- ID and shape 1.1 negative Enterobacter spp; Pantoea spp. Rod-shaped 1.2 negative Enterobacter spp; Pantoea spp. Rod-shaped 1.3 negative Enterobacter spp; Pantoea spp. Rod-shaped 2.1 negative Enterobacter spp; Pantoea spp. Rod-shaped 4.1 negative Enterobacter spp; Pantoea spp. Rod-shaped 4.2 negative Enterobacter spp; Klebsiella aerogenes. Rod-shaped Filtration: 6.1 negative Enterobacter spp; Klebsiella aerogenes. Rod-shaped 200 mL of water was filtered at a time using a vacuum filtration pump, shown below in Fig. 3. Filter papers of decreasing pore size (beginning with 10 L and ending at 0.6 L) were used to collect bacteria present. These filters were transferred to Mueller Hinton (MH) agar plates and incubated at either 37C or 30C for both 24 and 48 hours to isolate slow and fast-growing bacteria. 6.2 negative Enterobacter spp; Pantoea spp. Rod-shaped 13.2 negative Enterobacter spp; Leclercia adecarboxylata. Rod-shape 13.3 negative Enterobacter spp; Leclercia adecarboxylata. Rod-shape 15.3 negative Enterobacter spp; Leclercia adecarboxylata. Rod-shape 15.4 negative Enterobacter spp; Leclercia adecarboxylata. Rod-shape 16.2 negative Enterobacter spp; Leclercia adecarboxylata. Rod-shape Water Collection: Approximately 500 mL of water was collected using an autoclaved one-liter glass bottle. Water samples were obtained every two weeks from an overflow drain to minimize sediment. OBJECTIVE The main objective of this project was to gather pure bacterial isolates from the NMPE to further identify pathogenic gram-negative bacteria, test their resistance to a variety of antibiotics, and sequence the antibiotic resistant genes they contain. The results of this study will help create a resistance profile to serve as a guide for creating and implementing water management protocols. Water Collection Filtration and Incubation Colony Isolation Fig. 5. A small sample of isolates gathered and their identification Gram-Staining and Microscopy Isolation: After incubation, bacteria grown on the plates were streaked onto MacConkey Plates for isolation of gram-negative bacteria. Subculturing was repeated until isolated colonies were attained. Glycerol Stocks Gram-staining: The subculture plates were routinely examined using gram-staining to identify colony morphology and the progress in attaining pure isolates. Glycerol Stocks: Fig. 1. Map showing Nina Mason Pulliam EcoLab, the source of water collected. RESULTS Each isolate was inoculated in 2 mL of MH broth and placed in a shaking 37C incubator for 24 hours. 850 L of each culture was added to 150 L of sterilized glycerol in glycerol stock tubes. These tubes were frozen in the -80C freezer. Each sample was then streaked onto MH agar plates to be sent out for sequencing and identification. Fig. 3. The vacuum filtration pump used to collect bacteria from water samples v Fifty-five bacterial isolates were sent for sequencing. A sample of the resulting data is shown above in Fig. 5. v BLAST analysis of the isolates showed that the entirety of the isolates were gram-negative and members of the Enterobacteriaceae family of bacteria. v Enterobacter species got the highest scores in BLAST analysis but it also picked up other members of the Enterobacteriaceae family including Serratia, Citrobacter and Klebsiella v One possibility is that the location from which water was sampled had a high concentration of the Enterobacter species v A more precise ribosomal RNA homology search database will confirm the identity of the isolates DISCUSSION Send out for identification Fig. 2. Flow diagram giving an overview of the sampling and data collection procedure. v All the bacteria isolated belong to a family that has many human pathogens. For example: Klebsiella aerogenes, a subspecies of Klebsiella pneumoniae is a prominent pathogen of multi-drug resistance pneumonia that is responsible for hospital outbreaks worldwide Pantonea species have been identified as causative agents of infection in children and elderly and responsible for outbreaks of sepsis in NICUs Leclercia species have been reported as an opportunistic pathogen in immunocompromised hosts v The isolated bacterial samples obtained from the Ecolab are going to be evaluated for the degree of antibiotic resistance using Kirby Bauer assay v Resistance genes will be identified with PCR and sequence analysis and it will be determined whether they are expressed on a plasmid or the genome ACKNOWLEDGEMENTS We would like to thank Marian University for providing the laboratory, equipment, and facilities that allowed this research to be conducted. In addition, we would like to sincerely thank Dr. Ahmad and Dr. Akbar for advising and assisting us throughout this process.  ...