Synchrotron CT of an equine digit at the Australian Synchrotron Imaging and Medical Beamline

Laminitis is an extremely painful and debilitating condition of horses that can affect their athletic ability and even quality of life. The current gold standard for assessment of laminar tissue is histology, which is the only modality that enables detailed visualization of the lamina. Histology requires dissection of the hoof and therefore can only represent one specific time point. The superior spatial and contrast resolution of synchrotron computed tomography (sCT), when compared with readily available imaging modalities, such as radiographs and conventional CT, provides an opportunity for detailed studies of the lamina without the need for hoof dissection and histological assessment. If the resolution of histology can be matched or even approached, dynamic events, such as laminar blood flow, could also be studied on the microscopic tissue level. To investigate this possible application of sCT further, two objectives are presented: (i) to develop a protocol for sCT of an equine digit using cadaver limbs and (ii) to apply the imaging protocol established during (i) for sCT imaging of the vasculature within the foot using an ex vivo perfusion system to deliver the vascular contrast. The hypotheses were that sCT would allow sufficient resolution for detailed visualization to the level of the secondary lamellae and associated capillaries within the equine digit. Synchrotron CT enabled good visualization of the primary lamellae (average length 3.6 mm) and the ex vivo perfusion system was able to deliver vascular contrast agent to the vessels of the lamina. The individual secondary lamellae (average length 0.142 mm) could not be seen in detail, although differentiation between primary and secondary lamellae was achieved. This approaches, but does not yet reach, the current gold standard, histology, for assessment of the lamellae; however, with further refinement of this imaging technique, improved resolution may be accomplished in future studies.

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102 Functional and metabolic phenotyping of the murine heart using an ex vivo perfusion system and 13C-substrates

European Journal of Heart Failure Supplements ◽

10.1016/s1567-4215(03)90037-3 ◽

2003 ◽

Vol 2 (1) ◽

pp. 12

Author(s):

M KHAIRALLAH ◽

B BOUCHARD ◽

J MCDUFF ◽

F LABARTHE ◽

G DANIALOU ◽

...

Keyword(s):

Ex Vivo ◽

Perfusion System ◽

Metabolic Phenotyping ◽

Ex Vivo Perfusion

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Extrakorporális és intrakorporális anasztomózis rövid távú eredményeinek összehasonlítása malignus daganat miatt végzett laparoszkópos jobb oldali hemikolektómia esetén

Magyar Sebészet (Hungarian Journal of Surgery) ◽

10.1556/1046.73.2020.4.3 ◽

2020 ◽

Vol 73 (4) ◽

pp. 148-152

Author(s):

Kornél Vajda ◽

László Sikorszki

Keyword(s):

Length Of Stay ◽

Gold Standard ◽

Average Length ◽

Colon Surgery ◽

Right Hemicolectomy ◽

Malignant Tumours ◽

Short Term ◽

Significant Difference ◽

Laparoscopic Assisted ◽

Laparoscopic Hemicolectomy

Összefoglaló. Bevezetés: A laparoszkópia térhódítása a jobb oldali colon műtéteknél is nyilvánvaló. Ma legtöbb helyen a laparoszkóposan asszisztált jobb oldali hemikolektómia extrakorporális anasztomózissal a gold standard. A morbiditás randomizált vizsgálatok alapján még 30% körüli. A technikai fejlődés lehetővé tette az intrakorporális anasztomózist. Célkitűzés: Retrospektív módon elemezni rosszindulatú jobb oldali vastagbéldaganat miatt végzett laparoszkópos hemikolektómiák rövid távú eredményeit a két módszer összehasonlításával. Eredmények: 2018. 01. 01. – 2019. 12. 31. között 184 jobb oldali hemikolektómiát végeztünk, ezek közül 122 történt malignus betegség miatt. 51 esetben nyitott és 71 esetben laparoszkópos műtét történt. 37 férfi (átlagéletkor: 70,59 év) és 34 nő (átlagéletkor: 72,14 év) volt. 50 esetben extrakorporális (EA) és 21 esetben pedig intrakorporális anasztomózist (IA) végeztünk. Az EA csoportban 18, míg az IA csoportban 3 szövődmény alakult ki 30 napon belül (p = 0,067). Az EA csoportból 3, az IA csoportból 1 beteget veszítettünk el 30 napon belül (p = 0,66). Az átlagos ápolási idő az EA csoportban 9,48 (5–32) nap, míg az IA csoportban 6,52 (4–19) nap volt (p = 0,001) a szövődményes esetekkel együtt. A szövődményes esetek nélkül az EA csoportban 6,35 (5–10) nap, az IA csoportban pedig 5,55 (4–8) napnak bizonyult (p = 0,09). A műtéti idő pedig az EA csoportban 147 (90–240) perc, az IA csoportban pedig 146,47 (90–265) perc volt (p = 0,11). Konklúzió: Az irodalommal összhangban azt találtuk, hogy IA esetén kevesebb a szövődmény, ezzel is összefüggésben rövidebb az átlagos ápolási idő, és a műtéti időt tekintve nincs szignifikáns különbség. Ezeket figyelembe véve az intrakorporális anasztomózis javasolható jobb oldali laparoszkópos hemikolektómia esetén. Summary. Introduction: Laparoscopy became evident for right-sided colon surgery too. Today the laparoscopic-assisted right-hemicolectomy is the gold standard with extracorporeal anastomosis. Morbidity according to randomized trials is still approximately 30%. The development of the surgical technique resulted in the creation of intracorporeal anastomosis. Our aim was to compare the short-term results of the two methods. Aim: To analyse the short-term results of right-sided hemicolectomy that were performed due to malignant tumours with the comparison of the two methods. Results: A cohort of 184 right-sided hemicolectomy were performed from 01.01.2018 to 31.12.2019 from which 122 were operated on because of a malignant disease. 51 open and 71 laparoscopic operations were performed. The average age of 37 men and 34 women were 70.59 and 72.14 years, respectively. 50 patients underwent extracorporeal (EA) anastomosis and 21 intracorporeal (IA) anastomosis. Within 30 days the number of complications were 18 in the EA group and 3 in the IA group (p = 0.067). 3 from the EA group and 1 from IA group died within 30 days (p = 0.66). The average length of stay were 9.48 days in the EA group and 6.52 days in the IA group together with the complicated cases (p = 0.001) while 6.35 days and 5.55 days without the complicated cases (p = 0.09). The average duration of operation was 147 minutes in the EA and 146.47 minutes in the IA group (p = 0.11). Conclusion: We found concordance with the literature that there are fewer complications in case of IA which might be related to shorter length of stay. There is no significant difference between the surgical times. Bearing these facts in mind, IA might be suggested for right- sided laparoscopic hemicolectomy.

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Loss of an Intimin-Like Protein Encoded on a Uropathogenic E. coli Pathogenicity Island Reduces Inflammation and Affects Interactions with the Urothelium

Infection and Immunity ◽

10.1128/iai.00275-21 ◽

2021 ◽

Author(s):

Allyson E. Shea ◽

Jolie A. Stocki ◽

Stephanie D. Himpsl ◽

Sara N. Smith ◽

Harry L. T. Mobley

Keyword(s):

Ex Vivo ◽

Cell Adherence ◽

Upec Strain ◽

E Coli ◽

Bladder Cell ◽

Bladder Cells ◽

Strain Cft073 ◽

The Individual ◽

Tract Infections

Uropathogenic Escherichia coli (UPEC) causes the majority of uncomplicated urinary tract infections (UTI), which affect nearly half of women worldwide. Many UPEC strains encode an annotated intimin-like adhesin ( ila ) locus in their genome related to a well-characterized virulence factor in diarrheagenic E. coli pathotypes. Its role in UPEC uropathogenesis, however, remains unknown. In prototype UPEC strain CFT073, there is an ila locus that encodes three predicted intimin-like genes sinH , sinI , and ratA . We used in silico approaches to determine the phylogeny and genomic distribution of this locus among uropathogens. We found that the currently annotated intimin-encoding proteins in CFT073 are more closely related to invasin proteins found in Salmonella . Deletion of the individual sinH , sinI , and ratA genes did not result in measurable effects on growth, biofilm formation, or motility in vitro . On average, sinH was more highly expressed in clinical strains during active human UTI than in human urine ex vivo . Unexpectedly, we found that strains lacking this ila locus had increased adherence to bladder cells in vitro , coupled with a decrease in bladder cell invasion and death. The sinH mutant displayed a significant fitness defect in the murine model of ascending UTI including reduced inflammation in the bladder. These data confirmed an inhibitory role in bladder cell adherence to facilitate invasion and inflammation; therefore, the ila locus should be termed invasin-like, rather than intimin-like. Collectively, our data suggest that loss of this locus mediates measurable interactions with bladder cells in vitro and contributes to fitness during UTI.

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Lymphatic Vascular Structures: A New Aspect in Proliferative Diabetic Retinopathy

International Journal of Molecular Sciences ◽

10.3390/ijms19124034 ◽

2018 ◽

Vol 19 (12) ◽

pp. 4034 ◽

Cited By ~ 4

Author(s):

Erika Gucciardo ◽

Sirpa Loukovaara ◽

Petri Salven ◽

Kaisa Lehti

Keyword(s):

Diabetic Retinopathy ◽

Ex Vivo ◽

Tissue Level ◽

Clinical Samples ◽

Related Factors ◽

Disease Etiology ◽

Working Age ◽

Vascular Structures ◽

Underlying Mechanisms ◽

End Stage

Diabetic retinopathy (DR) is the most common diabetic microvascular complication and major cause of blindness in working-age adults. According to the level of microvascular degeneration and ischemic damage, DR is classified into non-proliferative DR (NPDR), and end-stage, proliferative DR (PDR). Despite advances in the disease etiology and pathogenesis, molecular understanding of end-stage PDR, characterized by ischemia- and inflammation-associated neovascularization and fibrosis, remains incomplete due to the limited availability of ideal clinical samples and experimental research models. Since a great portion of patients do not benefit from current treatments, improved therapies are essential. DR is known to be a complex and multifactorial disease featuring the interplay of microvascular, neurodegenerative, metabolic, genetic/epigenetic, immunological, and inflammation-related factors. Particularly, deeper knowledge on the mechanisms and pathophysiology of most advanced PDR is critical. Lymphatic-like vessel formation coupled with abnormal endothelial differentiation and progenitor cell involvement in the neovascularization associated with PDR are novel recent findings which hold potential for improved DR treatment. Understanding the underlying mechanisms of PDR pathogenesis is therefore crucial. To this goal, multidisciplinary approaches and new ex vivo models have been developed for a more comprehensive molecular, cellular and tissue-level understanding of the disease. This is the first step to gain the needed information on how PDR can be better evaluated, stratified, and treated.

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Case series: Transplantation of kidneys from donors with renal artery aneurysm

Canadian Urological Association Journal ◽

10.5489/cuaj.4244 ◽

2017 ◽

Vol 11 (7) ◽

pp. E307-10 ◽

Cited By ~ 3

Author(s):

Mahmoud Alameddine ◽

Zhobin Moghadamyeghaneh ◽

Giselle Guerra ◽

Mahmoud Morsi ◽

Mohammed Osman ◽

...

Keyword(s):

Renal Function ◽

Renal Artery ◽

Ex Vivo ◽

Average Length ◽

Graft Function ◽

Case Series ◽

Artery Aneurysm ◽

Renal Artery Aneurysm ◽

Antibody Mediated Rejection ◽

The Mean

Introduction: With the present disparity between organ availability and recipient demands, we reported our experience in transplanting kidneys with renal artery aneurysm after back-table reconstruction.Methods: Four patients were identified. The repair consisted of excision of the aneurysm with ostial closure, and for one of the cases, an ovarian vein patch was used. We reviewed the safety and outcomes of this procedure. All donors were asymptomatic before surgery and were diagnosed incidentally during living donor evaluation. The nephrectomies performed were hand-assisted laparoscopic approaches. All recipients had followup renal function and ultrasound duplex of renal artery at six and 12 months and then annually.Results: The mean age of the recipients was 28.7 years (range 3‒45). The mean size of the aneurysm was 7.4 ± 2.7 mm. All patients had immediate graft function with median serum creatinine of 1.9 ± 1.5 mg/dL at discharge. The average length of hospital stay was 6.25 ± 2.6 days. They also maintained good renal function with an average estimated glomerular filtration rate (eGFR) of 102.8 mL/min/1.73m2 (range 53.4‒199 mL/min/1.73m2) and patent vessels at one year. One patient suffered from acute antibody-mediated rejection and lost his graft (medication non-compliance). One patient had two simultaneous benign renal cysts that were resected. Three of the kidneys were right-sided and one left. Mean cold ischemia time was 86 ± 18 minutes. No deaths have been recorded.Conclusions: Transplanting kidneys with a renal artery aneurysm after ex-vivo repair is safe and the outcomes are encouraging. Also, it may play an important role in

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An individual-based mechanical model of cell movement in heterogeneous tissues and its coarse-grained approximation

10.1101/485276 ◽

2018 ◽

Cited By ~ 1

Author(s):

R. J. Murphy ◽

P. R. Buenzli ◽

R. E. Baker ◽

M. J. Simpson

Keyword(s):

Mechanical Model ◽

Numerical Solutions ◽

Cell Movement ◽

Biological Tissues ◽

Tissue Level ◽

Cellular Level ◽

Coarse Grained ◽

Mechanical Heterogeneity ◽

Single Scale ◽

The Individual

AbstractMechanical heterogeneity in biological tissues, in particular stiffness, can be used to distinguish between healthy and diseased states. However, it is often difficult to explore relationships between cellular-level properties and tissue-level outcomes when biological experiments are performed at a single scale only. To overcome this difficulty we develop a multi-scale mathematical model which provides a clear framework to explore these connections across biological scales. Starting with an individual-based mechanical model of cell movement, we subsequently derive a novel coarse-grained system of partial differential equations governing the evolution of the cell density due to heterogeneous cellular properties. We demonstrate that solutions of the individual-based model converge to numerical solutions of the coarse-grained model, for both slowly-varying-in-space and rapidly-varying-in-space cellular properties. Applications of the model are discussed, including determining relative cellular-level properties and an interpretation of data from a breast cancer detection experiment.

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Surface modification of decellularized bovine carotid arteries with human vascular cells significantly reduces their thrombogenicity

Journal of Biological Engineering ◽

10.1186/s13036-021-00277-2 ◽

2021 ◽

Vol 15 (1) ◽

Author(s):

Eriselda Keshi ◽

Peter Tang ◽

Marie Weinhart ◽

Hannah Everwien ◽

Simon Moosburner ◽

...

Keyword(s):

Surface Modification ◽

Whole Blood ◽

Carotid Arteries ◽

Complete Blood Count ◽

Ex Vivo ◽

Vascular Grafts ◽

Blood Perfusion ◽

Vascular Cells ◽

Perfusion System ◽

Human Whole Blood

Abstract Background Since autologous veins are unavailable when needed in more than 20% of cases in vascular surgery, the production of personalized biological vascular grafts for implantation has become crucial. Surface modification of decellularized xenogeneic grafts with vascular cells to achieve physiological luminal coverage and eventually thromboresistance is an important prerequisite for implantation. However, ex vivo thrombogenicity testing remains a neglected area in the field of tissue engineering of vascular grafts due to a multifold of reasons. Methods After seeding decellularized bovine carotid arteries with human endothelial progenitor cells and umbilical cord-derived mesenchymal stem cells, luminal endothelial cell coverage (LECC) was correlated with glucose and lactate levels on the cell supernatant. Then a closed loop whole blood perfusion system was designed. Recellularized grafts with a LECC > 50% and decellularized vascular grafts were perfused with human whole blood for 2 h. Hemolysis and complete blood count evaluation was performed on an hourly basis, followed by histological and immunohistochemical analysis. Results While whole blood perfusion of decellularized grafts significantly reduced platelet counts, platelet depletion from blood resulting from binding to re-endothelialized grafts was insignificant (p = 0.7284). Moreover, macroscopic evaluation revealed thrombus formation only in the lumen of unseeded grafts and histological characterization revealed lack of CD41 positive platelets in recellularized grafts, thus confirming their thromboresistance. Conclusion In the present study we were able to demonstrate the effect of surface modification of vascular grafts in their thromboresistance in an ex vivo whole blood perfusion system. To our knowledge, this is the first study to expose engineered vascular grafts to human whole blood, recirculating at high flow rates, immediately after seeding.

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