Cord Lining Mesenchymal Stem Cells Have a Modest Positive Effect on Angiogenesis in Hindlimb Ischemia

Purpose: We investigated the use of human Cord Lining Mesenchymal Stem Cells (CL-MSCs) (US Patent number 9,737,568), in a rabbit hindlimb ischemia model, and evaluated their potential in stimulating neovascularization. Allogenic human CL- MSCs could potentially be used to treat patients with lower limb ischemia and non-healing wounds.Methods: Twenty rabbits were divided into two separate groups. We created a hindlimb ischemia model surgically. At 21 and 49 days post-operatively, animals in the treatment group were injected with CL-MSCs (500,000 cells per 0.2 ml on each site) at 10 different sites (Quadriceps- 4 sites, Hamstrings- 4 sites and Calf-−2 sites) in the hindlimb muscles. The control group received only saline injection to the corresponding sites at the same time point as the treatment group. We then evaluated the effects of treatment on neovascularization by angiography, laser doppler perfusion imaging, as well as by histology. We evaluated the tissue samples for any signs of local immune reaction to the cell implantation. We also observed the rabbit clinically for any adverse effects after treatment.Results: We found a higher number of CD31 positive cells in the treatment group, with a greater number of capillaries found in the treated muscles. The Rectus Femoris demonstrated a median vessel count/muscle fiber of 0.121 for the treatment group, compared to 0.076 in the control group (median difference 0.04; 95% CI 0.001–0.11; p = 0.041). The Gastrocnemius demonstrated a median vessel count/muscle fiber of 0.175 for the treatment group, compared to 0.089 in the control group (median difference 0.087; 95% CI −0.006 to 0.234; p = 0.07). Blood perfusion quantification through Laser Doppler Perfusion Imaging (LDPI) also demonstrated a non-statistically significant increase in perfusion in favor of the treatment group. CL-MSCs demonstrated no toxicity associated morbidity and minimal local immune reaction to implantation.Conclusion: CL-MSCs have a positive effect on angiogenesis in a rabbit hindlimb ischemia model. This preliminary data is encouraging and paves the way for future large animal studies or for clinical trials.

Predicting Global Ship Demolition Using Machine Learning Approach

Global ship demolition is mostly concentrated in south Asian countries, namely Bangladesh, India, Pakistan and China, since 1990’s, having competitive advantage for their high natural tide, and low environmental and social costs. Due to high social and environmental externalities, stakeholders increase monitoring of the externalities and continue to prescribe improvement towards sustainability, which put pressures on profitability and competitiveness. As a consequence, also seen in the past, a leakage effect may emerge, leading to shift of this activity to a region, with relatively less monitored and less stricter on social and environmental impacts. Unfortunately, the leakage effect is never predicted in shipbreaking in order to understand the level of push compatible in the given socio-economic contexts. In this study, we have attempted to predict the future ship demolition landscape, applying machine learning technique to 34,531 in-service vessels worldwide, larger than 500 gross tonnage (GT), which is run against a learning model based on 3500 demolished vessels from 2014. This study shows that redistribution may occur among the top recycling nations: India may emerge out to be a dominant player in shipbreaking, surpassing Bangladesh by a margin of two-fold, while Pakistan and China are in decreasing trend. In addition, the leakage effect is observed, in that Vietnam is predicted to be the fourth largest ship demolition country, while China and Pakistan recede from the third and fourth place to 6th and 8th. Turkey is predicted to advance from fifth position to third position by vessel count but stays same in term of total GT dismantled. Although it is not clear if any leakage is to be observed in the near future, this study may be a model for future predictive analytics and help stakeholders take evidence-based business decisions.

B-Flow Sonography vs. Color Doppler Sonography for the Assessment of Vascularity in Pediatric Kidney Transplantation

Objective To compare B-flow sonography (BFS) with color Doppler sonography (CDS) for imaging of kidney transplant vascularization in children. Patients and Methods All children receiving a kidney transplantation who underwent a protocol-based ultrasound examination (Loqiq 9, GE Medical Systems, Milwaukee, WI, USA) using the BFS and CDS technique with equal settings and probe position between January 2013 and January 2016 were retrospectively assessed (n = 40). The obtained datasets were visually graded according to the following criteria: (I) delineation of the renal vascular tree (Grade 1 – clear demarcation of interlobar, together with arcuate and interlobular vessels; Grade 2 – clear demarcation of interlobar and cortical vessels, but no distinction of interlobular from arcuate vessels; Grade 3 – only clear demarcation of interlobar vessels, Grade 4 – insufficient demarcation) (II) delineation of cortical vessel density in ventral, lateral, and dorsal part of the transplant, (III) smallest vessel-capsule distance, and (IV) maximum cortical vessel count. Comparison between methods was performed using Fisher’s exact and paired sample t-tests. Results Applying a curved transducer (C1–6), BFS showed superior delineation of the renal vascular tree (p < 0.001), a lower vessel-capsule distance (p < 0.001), a higher cortical vessel count (p < 0.001), and a higher cortical vessel density in the superficial cortex (p = 0.01) than CDS. In the dorsal and lateral aspects of the transplant, cortical vessel density was lower with BFS (both p < 0.001). Using a linear high-resolution transducer (ML 6–15), no significant differences between the methods were found. Conclusion Improved imaging of kidney transplant vascularization can be achieved in children by adding BFS to a standard protocol. The BFS technique is especially beneficial for overall assessment of the renal vascular tree together with the extent of cortical vascularization on curved array images. Key points: Citation Format

Value of contrast-enhanced ultrasound for detection of synovial vascularity in experimental rheumatoid arthritis: an exploratory study

Objective This study aimed to examine the associations between contrast-enhanced ultrasound (CEUS) imaging and synovial hypervascularity and synovitis score in a rabbit model of antigen-induced arthritis (AIA), compared with power Doppler ultrasound (PDUS). Methods We investigated 50 knee joints in 25 AIA rabbits (AIA group), and 10 knee joints in five sham-injected rabbits (control group). PDUS and CEUS images were evaluated at the 8th week. Ultrasound-guided synovial biopsies were targeted in the area with hypervascularity, and synovial microvessel density (MVD) was evaluated by immunohistochemical staining of CD31. Results The PDUS score was significantly higher in the AIA group (2.61 ± 0.78) compared with the control group (0.50 ± 0.53). CEUS in the AIA group revealed a fast-in/slow-out pattern of contrast enhancement. MVD revealed by CD31+ vessel count and the synovitis score were significantly higher in the AIA group compared with the control group. In the AIA group, CEUS findings showed a better correlation with MVD revealed by CD31+ and synovitis score than PDUS findings. Conclusion CEUS is superior to PDUS for estimating synovial hypervascularity and hyperplasia in experimental rheumatoid arthritis.

Pro-Fibrogenic and Anti-Inflammatory Potential of a Polyphenol-Enriched Fraction from Annona crassiflora in Skin Repair

A polyphenol-enriched fraction from Annona crassiflora fruit peel (PEF-Ac) containing chlorogenic acid, epi-catechin, procyanidins B2 and C1, quercetin-glucoside, kaempferol, and caffeoyl-glucoside was investigated for its anti-inflammatory, pro-angiogenic, and profibrogenic potential in the healing of cutaneous wounds. Four wounds were performed on the back of C57 mice and the lesions were treated with the vehicle (Vaseline and lanolin) and PEF-Ac at concentrations of 2%, 4%, and 6% for 4 and 7 d. Neutrophils and macrophages activities were evaluated indirectly by the activity of myeloperoxidase and N-acetyl-β-D-glycosaminidase, angiogenesis was evaluated by hemoglobin dosing and vessel count in histological sections, and collagen deposition was assessed from histological sections stained with picrosirius red. PEF-Ac demonstrated anti-inflammatory activity, with reduced activities of neutrophil and macrophage in the cutaneous wounds. In addition, there was an increase in the synthesis of types I and III collagen, as well as in the percentage of wound closure, mainly after 4 d of treatment. On the other hand, PEF-Ac did not present an effective pro-angiogenic activity. A. crassiflora fruit peel showed anti-inflammatory and profibrogenic properties, indicating a promising natural source of bioactive molecules for treatment of cutaneous wounds.

Abstract TMP111: Thalidomide and Lenalidomide Treatment Reduces Microhemorrhage in Brain Arteriovenous Malformation in Mice Through Increasing Mural Cell Coverage

Introduction: Brain arteriovenous malformations (bAVMs) have an abnormal vessel wall and are prone to rupture. The mechanism of bAVM rupture is unclear. In Alk1 -deficient mice, bAVM vessels have fewer mural cells. In endoglin-deficient mice, thalidomide increases mural cells in retina AVM vessels. We hypothesize that thalidomide and its less toxic analogue, lenalidomide, improve vessel mural cell coverage and reduce microhemorrhage in Alk1 -deficient bAVM. Methods: Brain AVMs were induced in adult Alk1 2f/2f mice through induction of focal Alk1 gene deletion and angiogenic stimulation. Thalidomide was injected intraperitoneally (i.p.) twice per week for six weeks, starting either 2 weeks after model induction when bAVMs were beginning to develop or 8 weeks after when bAVMs were fully developed. Lenalidomide treatment was started 8 weeks after model induction through i.p. injection daily for six weeks. Results: Thalidomide treatment starting 2 weeks after bAVM induction reduced the number of abnormal vessels and microhemorrhage and increased vascular smooth muscle (vSM)-coverage. Thalidomide also increased the expression of platelet-derived growth factor b (pdgfb) and its receptor (pdgfr beta), indicating that pdgfg/pdgfr beta signaling is one of the mechanisms responsible for the improvement of mural cell coverage. Thalidomide and lenalidomide treatment started at the later time point also improved vSM-coverage and showed a trend toward reduction of microhemorrhage and abnormal vessel count. Conclusions: Thalidomide and lenalidomide stabilize the bAVM vessel wall and reduce microhemorrahge. Further studies are needed to determine whether these agents have a possible therapeutic value for patients.

Fetal blood vessel count increases in compensation of hypoxia in premature placentas

BACKGROUND Prematurity refers to live births before 37 weeks of gestation, wherein the baby is born before the body and its organ systems achieve perfect maturity, and this disorder is still a global problem. The high incidence of prematurity is a problem in developing and also in developed countries. Certain conditions accompanying pregnancies like preeclampsia, infection, and placental insufficiency, may trigger uterine hypoxia, causing premature birth. The placental condition is related to the intra-uterine fetal condition. In prolonged placental hypoxia, there occurs a compensatory mechanism, i.e. an increase in placental angiogenesis. This study aimed to evaluate the effect of hypoxia on fetal blood vessel count as compensatory mechanism for tissue hypoxia. METHODS An observational-analytical cross-sectional design using paraffin blocks of conserved premature placentas, comprising 31 samples of hypoxic premature placentas and 28 samples of non-hypoxic premature placentas, selected using non-random consecutive sampling. The samples were made into slides and stained with hematoxylin-eosin for assessment of histological structure, including fetal blood vessel count and integrity, villus conditions, syncytiotrophoblastic nuclear changes, and syncytiotrophoblastic nuclear aggregation. Mann-Whitney test was used to compare the difference of blood vessel count between groups. RESULTS Assessment of histological structure showed a significant increase in fetal blood vessel count in the hypoxic group [8.00 (5-15)] as compared with the non-hypoxic group [7.50 (3-15)]. CONCLUSION The hypoxia in premature placentas caused an increase in the number of fetal blood vessels as a form of compensation for disturbed oxygen homeostasis.