Search

Metastasis via the lymphatic vasculature is an important step in cancer progression. The formation of new lymphatic vessels (lymphangiogenesis), or remodeling of existing lymphatics, is thought to facilitate the entry and transport of tumor cells into lymphatic vessels and on to distant organs. The migration of lymphatic endothelial cells (LEC) toward guidance cues is critical for lymphangiogenesis. While chemokines are known to provide directional navigation for migrating immune cells, their role in mediating LEC migration during tumor-associated lymphangiogenesis is not well defined. Here, we undertook gene profiling studies to identify chemokine–chemokine receptor pairs that are involved in tumor lymphangiogenesis associated with lymph node metastasis. CCL27 and CCL28 were expressed in tumor cells with metastatic potential, while their cognate receptor, CCR10, was expressed by LECs and upregulated by the lymphangiogenic growth factor VEGFD and the proinflammatory cytokine TNFα. Migration assays demonstrated that LECs are attracted to both CCL27 and CCL28 in a CCR10-dependent manner, while abnormal lymphatic vessel patterning in CCR10-deficient mice confirmed the significant role of CCR10 in lymphatic patterning. In vivo analyses showed that LECs are recruited to a CCL27 or CCL28 source, while VEGFD was required in combination with these chemokines to enable formation of coherent lymphatic vessels. Moreover, tumor xenograft experiments demonstrated that even though CCL27 expression by tumors enhanced LEC recruitment, the ability to metastasize was dependent on the expression of VEGFD. These studies demonstrate that CCL27 and CCL28 signaling through CCR10 may cooperate with inflammatory mediators and VEGFD during tumor lymphangiogenesis.Significance:The study shows that the remodeling of lymphatic vessels in cancer is influenced by CCL27 and CCL28 chemokines, which may provide a future target to modulate metastatic spread.

Supplementary Figures S1-S8 S1 Heat map of chemokines and receptors S2 Immunohistochemical staining of lymphatic vessels S3 Expression of VEGF-D and chemokines in cancer cell lines S4 Expression of genes in melanoma patient samples S5 Chemokine secretion by tumour cells S6 Chemokine induced migration of LEC S7 Chemokines attract LEC in vivo S8 CCL-293EBNA tumour growth and CCL27 secretion

PDF file - 147K

PDF file - 86K

PDF file - 162K

Lymph node metastasis, an early and prognostically important event in the progression of many human cancers, is associated with expression of VEGF-D. Changes to lymph node vasculature that occur during malignant progression may create a metastatic niche capable of attracting and supporting tumor cells. In this study, we sought to characterize molecules expressed in lymph node endothelium that could represent therapeutic or prognostic targets. Differential mRNA expression profiling of endothelial cells from lymph nodes that drained metastatic or nonmetastatic primary tumors revealed genes associated with tumor progression, in particular bone morphogenetic protein-4 (BMP-4). Metastasis driven by VEGF-D was associated with reduced BMP-4 expression in high endothelial venules, where BMP-4 loss could remodel the typical high-walled phenotype to thin-walled vessels. VEGF-D expression was sufficient to suppress proliferation of the more typical BMP-4–expressing high endothelial venules in favor of remodeled vessels, and mechanistic studies indicated that VEGF receptor-2 contributed to high endothelial venule proliferation and remodeling. BMP-4 could regulate high endothelial venule phenotype and cellular function, thereby determining morphology and proliferation responses. Notably, therapeutic administration of BMP-4 suppressed primary tumor growth, acting both at the level of tumor cells and tumor stromal cells. Together, our results show that VEGF-D–driven metastasis induces vascular remodeling in lymph nodes. Furthermore, they implicate BMP-4 as a negative regulator of this process, suggesting its potential utility as a prognostic marker or antitumor agent. Cancer Res; 71(20); 6547–57. ©2011 AACR.

PDF file - 98K

PDF file - 161K

PDF file - 79K

Objectives Lipedema, a poorly understood chronic disease of adipose hyper-deposition, is often mistaken for obesity and causes significant impairment to mobility and quality-of-life. To identify molecular mechanisms underpinning lipedema, we employed comprehensive omics-based comparative analyses of whole tissue, adipocyte precursors (adipose-derived stem cells (ADSCs)), and adipocytes from patients with or without lipedema. Methods We compared whole-tissues, ADSCs, and adipocytes from body mass index–matched lipedema (n = 14) and unaffected (n = 10) patients using comprehensive global lipidomic and metabolomic analyses, transcriptional profiling, and functional assays. Results Transcriptional profiling revealed >4400 significant differences in lipedema tissue, with altered levels of mRNAs involved in critical signaling and cell function-regulating pathways (e.g., lipid metabolism and cell-cycle/proliferation). Functional assays showed accelerated ADSC proliferation and differentiation in lipedema. Profiling lipedema adipocytes revealed >900 changes in lipid composition and >600 differentially altered metabolites. Transcriptional profiling of lipedema ADSCs and non-lipedema ADSCs revealed significant differential expression of >3400 genes including some involved in extracellular matrix and cell-cycle/proliferation signaling pathways. One upregulated gene in lipedema ADSCs, Bub1, encodes a cell-cycle regulator, central to the kinetochore complex, which regulates several histone proteins involved in cell proliferation. Downstream signaling analysis of lipedema ADSCs demonstrated enhanced activation of histone H2A, a key cell proliferation driver and Bub1 target. Critically, hyperproliferation exhibited by lipedema ADSCs was inhibited by the small molecule Bub1 inhibitor 2OH-BNPP1 and by CRISPR/Cas9-mediated Bub1 gene depletion. Conclusion We found significant differences in gene expression, and lipid and metabolite profiles, in tissue, ADSCs, and adipocytes from lipedema patients compared to non-affected controls. Functional assays demonstrated that dysregulated Bub1 signaling drives increased proliferation of lipedema ADSCs, suggesting a potential mechanism for enhanced adipogenesis in lipedema. Importantly, our characterization of signaling networks driving lipedema identifies potential molecular targets, including Bub1, for novel lipedema therapeutics.

Background Lymphoedema is an incurable and progressive disease that affects not only physical function but overall quality of life. Surgical treatment options for the management of lymphoedema are being increasingly performed. This study aims to review post-operative health-related quality of life (HRQOL) following surgical treatment of lymphoedema. Methods A systematic search of the PubMed and Medline databases was performed from the date of their inception until September 2018 to evaluate HRQOL following different surgical options for the treatment of lymphoedema. Results One hundred and thirteen articles were identified. Twenty-one articles were included in the final review, comprising a total of 736 patients. HRQOL improvements appear to be sustained for at least 6-12 months post-operatively. In particular, major benefits were noted in the domains based around physical functioning. Patient satisfaction similarly mirrors HRQOL improvements, following an initial dip in the immediate post-operative period. Conclusion All surgical treatment modalities for the management of lymphoedema confer significant HRQOL improvements across a diverse range of health domains, with this critical outcome of surgery an important pre-operative consideration. Recommendations for ongoing research are suggested.

Background: The present study assess lifestyle and eating habits of Iranian adolescent girls. Methods: This cross-sectional study was conducted on 3207 adolescent girls, aged 12–18 years. Lifestyle and eating habits of adolescents were assessed. Results: The prevalence of overweight was 11.3% and that of obesity was 10.2%. The frequencies of daily intake of fruits, dairy products and vegetables were 60.9%, 45.6 % and 33.5%, respectively. Skipping breakfast (15.8%) was more frequent than skipping dinner or lunch. Moreover, 10.6% of subjects had fast food daily and 16.2% had fast food weekly. Overall, 55.2% of participants watched television for at least 2 hours a day; and 27.1% reported to have a sleeping time of less than 8 hours a day. Low physical activity, i.e. less than 30 minutes per day, was less frequent among participants with high socio-economic status (SES) compared to those with medium or low SES (57.6% vs. 61.3% and 64.1%, respectively, P value = 0.010). Conclusion: Improvement of lifestyle habits should be considered in public health plans for health promotion of adolescent girls and the next generation. Primordial prevention of chronic diseases by improving healthy lifestyle of adolescent girls should be a national public health priority

BACKGROUND: The anterolateral thigh flap is a workhorse reconstructive flap. Versatility in design is a key strength but perforator anatomy can be variable. Inability to locate perforators prompts consideration of contralateral thigh exploration. However, such exploration would be futile if the absence of perforators proves symmetrical. This study assesses the symmetry of anterolateral thigh flap vasculature using computed tomography angiography (CTA). METHODS: A retrospective analysis of 20 bilateral thigh CTAs was performed. Each limb was assessed for number, course, location and size of perforators. Only vessels >0.5 mm in size at origin were included. Location was standardized between patients using perforator distance/thigh length ratio. Results were analysed using Wilcoxon signed-rank test. RESULTS: In each thigh, the average number of perforators was 3.58 and average perforator distance/thigh length ratio was 0.358 ± 0.08. Between both limbs of the same patient, the mean difference in number of perforators was 0.55 (P = 0.002), and difference in average perforator size was 0.3 mm (P < 0.001). Average perforator location differed by a mean of 3% of thigh length (P < 0.001) between thighs. CONCLUSION: While average vessel size and location appear similar, there does not appear to be symmetry in the number of perforators. Surgical exploration of the contralateral thigh in an absence of perforators should be considered. In patients where abnormal anatomy is expected, mapping with CTA could be considered to reduce morbidity associated with unsuccessful surgical exploration and dissection.

Lipedema, a poorly understood chronic disease of adipose hyper-deposition, is often mistaken for obesity and causes significant impairment to mobility and quality-of-life. To identify molecular mechanisms underpinning lipedema, we employed comprehensive omics-based comparative analyses of whole tissue, adipocyte precursors (adipose-derived stem cells (ADSCs)), and adipocytes from patients with or without lipedema.We compared whole-tissues, ADSCs, and adipocytes from body mass index-matched lipedema (n = 14) and unaffected (n = 10) patients using comprehensive global lipidomic and metabolomic analyses, transcriptional profiling, and functional assays.Transcriptional profiling revealed4400 significant differences in lipedema tissue, with altered levels of mRNAs involved in critical signaling and cell function-regulating pathways (e.g., lipid metabolism and cell-cycle/proliferation). Functional assays showed accelerated ADSC proliferation and differentiation in lipedema. Profiling lipedema adipocytes revealed900 changes in lipid composition and600 differentially altered metabolites. Transcriptional profiling of lipedema ADSCs and non-lipedema ADSCs revealed significant differential expression of3400 genes including some involved in extracellular matrix and cell-cycle/proliferation signaling pathways. One upregulated gene in lipedema ADSCs, Bub1, encodes a cell-cycle regulator, central to the kinetochore complex, which regulates several histone proteins involved in cell proliferation. Downstream signaling analysis of lipedema ADSCs demonstrated enhanced activation of histone H2A, a key cell proliferation driver and Bub1 target. Critically, hyperproliferation exhibited by lipedema ADSCs was inhibited by the small molecule Bub1 inhibitor 2OH-BNPP1 and by CRISPR/Cas9-mediated Bub1 gene depletion.We found significant differences in gene expression, and lipid and metabolite profiles, in tissue, ADSCs, and adipocytes from lipedema patients compared to non-affected controls. Functional assays demonstrated that dysregulated Bub1 signaling drives increased proliferation of lipedema ADSCs, suggesting a potential mechanism for enhanced adipogenesis in lipedema. Importantly, our characterization of signaling networks driving lipedema identifies potential molecular targets, including Bub1, for novel lipedema therapeutics.

The article's abstract is not available.

Objective: This study aims to assess the relationship of late-night cell phone use with sleep duration and quality in a sample of Iranian adolescents. Methods: The study population consisted of 2400 adolescents, aged 12–18 years, living in Isfahan, Iran. Age, body mass index, sleep duration, cell phone use after 9 p.m., and physical activity were documented. For sleep assessment, the Pittsburgh Sleep Quality Index questionnaire was used. Results: The participation rate was 90.4% (n = 2257 adolescents). The mean (SD) age of participants was 15.44 (1.55) years; 1270 participants reported to use cell phone after 9 p.m. Overall, 56.1% of girls and 38.9% of boys reported poor quality sleep, respectively. Wake-up time was 8:17 a.m. (2.33), among late-night cell phone users and 8:03 a.m. (2.11) among non-users. Most (52%) late-night cell phone users had poor sleep quality. Sedentary participants had higher sleep latency than their peers. Adjusted binary and multinomial logistic regression models showed that late-night cell users were 1.39 times more likely to have a poor sleep quality than non-users (p-value

Background/Objectives Sunlight is a major risk factor for cutaneous melanoma. However, its interaction with melanoma is complex. In particular, vitamin D is a UVB-derived hormone that has been shown to have anti-cancer effects. In this retrospective pilot study we sought to determine an association between the clinicopathological features of melanoma and the patients’ corresponding serum vitamin D level. Methods In total, 109 primary melanomas diagnosed between 2001 and 2013 were retrospectively identified from our institutional database with a corresponding 25-hydroxyvitamin D3 level estimated within 6 months of diagnosis. Tumour, clinical (age, sex, tumour location) and pathological (thickness, mitosis, ulceration, Clark level, subtype, metastatic status) parameters were correlated with vitamin D. For statistical analysis, an unpaired Student's t-test and anova was used for categorical variables, and Spearman's correlation for continuous variables. Results Vitamin D level was inversely associated with Breslow thickness as a dichotomous, categorical and continuous variable. The association remained significant when controlled for patient's age and sex (P = 0.026). Vitamin D was higher in non-ulcerated tumours compared with ulcerated tumours (P = 0.006) and in tumours with mitotic rate

In this video we describe our method for performing a robotic abdominoperineal resection, posterior vaginectomy and distal sacrectomy. We present the case of a 75y female with a recurrence of her rectal cancer on a background of extensive abdominal operations. She was initially diagnosed in 2014 at which time she underwent chemoradiotherapy. This was followed by two liver resections and a Hartmann's procedure. She received radiotherapy in 2018 for a metastatic right upper lobe lung lesion. She has also had a hysterectomy and cholecystectomy and has hypothyroidism and hypertension.

Background This study aims to systematically review the effects of maternal vitamin and/or mineral supplementation on the content of breast milk. Methods We systematically searched electronic databases including Medline via PubMed, Scopus and ISI Web of Science till May 24, 2018. The following terms were used systematically in all mentioned databases: (“human milk” OR “breast milk” OR “breast milk composition” OR “human breast milk composition” OR “composition breast milk” OR “mother milk” OR “human breast milk” OR “maternal milk”) AND (“vitamin a” OR “retinol” OR “retinal” OR “retinoic acid” OR “beta-carotene” OR “beta carotene” OR “ascorbic acid” OR “l-ascorbic acid” OR “l ascorbic acid” OR “vitamin c” OR “vitamin d” OR “cholecalciferol” OR “ergocalciferol” OR “calciferol” OR “vitamin e” OR “tocopherol” OR “tocotrienol” OR “alpha-tocopherol” OR “alpha tocopherol” OR “α-tocopherol” OR “α tocopherol” OR “vitamin k” OR “vitamin b” OR “vitamin b complex” OR “zinc” OR “iron” OR “copper” Or “selenium” OR “manganese” OR “magnesium”) and we searched Medline via Medical subject Headings (MeSH) terms. We searched Google Scholar for to increase the sensitivity of our search. The search was conducted on human studies, but it was not limited to the title and abstract. Methodological quality and risk of bias of included studies were evaluated by Jadad scale and Cochrane risk of bias tools, respectively. Results This review included papers on three minerals (zinc, iron, selenium) and 6 vitamins (vitamin A, B, D, C, E and K) in addition to multi-vitamin supplements. Although studies had different designs, e.g. not using random allocation and/or blinding, our findings suggest that maternal use of some dietary supplements, including vitamin A, D, vitamin B1, B2 and vitamin C might be reflected in human milk. Vitamin supplements had agreater effect on breast milk composition compared to minerals. Higher doses of supplements showed higher effects and they were reflected more in colostrum than in the mature milk. Conclusion Maternal dietary vitamin and/or mineral supplementation, particularly fat- soluble vitamins, vitamin B1, B2 and C might be reflected in the breast milk composition. No difference was found between mega dose and single dose administration of minerals.

Supplemental Digital Content is available in the text., Background: Cancer patients often require radiotherapy (RTx) to enhance their survival. Unfortunately, RTx also damages nearby healthy non-cancer tissues, leading to progressive fibrotic soft-tissue injury, consisting of pain, contracture, tissue-breakdown, infection, and lymphoedema. Mechanisms underlying the clinically observed ability of fat grafting to ameliorate some of these effects, however, are poorly understood. It was hypothesized that RTx significantly alters fibroblast cell function and the paracrine secretome of adipose-derived stem cells (ADSC) may mitigate these changes. Methods: To investigate cellular changes resulting in the fibrotic side-effects of RTx, cultured normal human dermal fibroblasts (NHDF) were irradiated (10Gy), then studied using functional assays that reflect key fibroblast functions, and compared with unirradiated controls. RNA-Seq and targeted microarrays (with specific examination of TGFβ) were performed to elucidate altered gene pathways. Finally, conditioned-media from ADSC was used to treat irradiated fibroblasts and model fat graft surgery. Results: RTx altered NHDF morphology, with cellular functional changes reflecting transition into a more invasive phenotype: increased migration, adhesion, contractility, and disordered invasion. Changes in genes regulating collagen and MMP homeostasis and cell-cycle progression were also detected. However, TGFβ was not identified as a key intracellular regulator of the fibroblast response. Finally, treatment with ADSC-conditioned media reversed the RTx-induced hypermigratory state of NHDF. Conclusions: Our findings regarding cellular and molecular changes in irradiated fibroblasts help explain clinical manifestations of debilitating RTx-induced fibrosis. ADSC-secretome-mediated reversal indicated that these constituents may be used to combat the devastating side-effects of excessive unwanted fibrosis in RTx and other human fibrotic diseases.

Metastasis via the lymphatic vasculature is an important step in cancer progression. The formation of new lymphatic vessels (lymphangiogenesis), or remodeling of existing lymphatics, is thought to facilitate the entry and transport of tumor cells into lymphatic vessels and on to distant organs. The migration of lymphatic endothelial cells (LEC) toward guidance cues is critical for lymphangiogenesis. While chemokines are known to provide directional navigation for migrating immune cells, their role in mediating LEC migration during tumor-associated lymphangiogenesis is not well defined. Here, we undertook gene profiling studies to identify chemokine–chemokine receptor pairs that are involved in tumor lymphangiogenesis associated with lymph node metastasis. CCL27 and CCL28 were expressed in tumor cells with metastatic potential, while their cognate receptor, CCR10, was expressed by LECs and upregulated by the lymphangiogenic growth factor VEGFD and the proinflammatory cytokine TNFα. Migration assays demonstrated that LECs are attracted to both CCL27 and CCL28 in a CCR10-dependent manner, while abnormal lymphatic vessel patterning in CCR10-deficient mice confirmed the significant role of CCR10 in lymphatic patterning. In vivo analyses showed that LECs are recruited to a CCL27 or CCL28 source, while VEGFD was required in combination with these chemokines to enable formation of coherent lymphatic vessels. Moreover, tumor xenograft experiments demonstrated that even though CCL27 expression by tumors enhanced LEC recruitment, the ability to metastasize was dependent on the expression of VEGFD. These studies demonstrate that CCL27 and CCL28 signaling through CCR10 may cooperate with inflammatory mediators and VEGFD during tumor lymphangiogenesis. Significance: The study shows that the remodeling of lymphatic vessels in cancer is influenced by CCL27 and CCL28 chemokines, which may provide a future target to modulate metastatic spread.

Background Free vascularized bone flaps are widely recognized as the optimal reconstruction for patients who undergo mandibular resection. The fibula-free flap is currently considered the gold standard, workhorse flap for mandibular reconstruction. Although previous studies have analysed individual success of each flap type, few have compared iliac- and fibula-free flaps. Methods A systematic review of the literature was conducted in line with the PRISMA protocol searching the PubMed and EBSCO databases. Twenty-four studies were included as per predetermined inclusion criteria. Double-arm random effect meta-analysis was conducted with STATA 12, and single-arm meta-analysis was conducted utilizing Meta-XL. Results The results of this meta-analysis confirm that there is no significant difference in total flap loss between fibula- and iliac-free flap reconstruction of mandibular defects. In terms of recipient site complication, there was a significantly higher risk of delayed healing and suture line breakdown in the iliac flap group (P = 0.05). Donor site complications showed a trend towards being higher in the fibula flap group. Osseointegrated dental implant loss in fibula flaps was higher than in iliac flaps (5.3% compared with 1.7%). Conclusion Both iliac- and fibula-free flaps should be considered for use in mandibular reconstruction. We suggest the iliac crest as the first choice for mandibular angle or body defects (better contour match) or also defects requiring greater soft-tissue bulk for intra-oral lining. The fibula flap is best when bony length is required such as in subtotal or total mandibulectomy.

The benefits of adult stem cells for repair of the heart have been attributed to the repertoire of salutary paracrine activities they appear to exert. We previously isolated human W8B2+ cardiac stem cells (CSCs) and found they powerfully influence cardiomyocytes and endothelial cells to collectively promote cardiac repair and regeneration. Here, the complexity of the W8B2+ CSC secretomes was characterised and examined in more detail. Using ion exchange chromatography to separate soluble proteins based on their net surface charge, the secreted factors responsible for the pro-survival activity of W8B2+ CSCs were found within the low and medium cation fractions. In addition to the soluble proteins, extracellular vesicles generated from W8B2+ CSCs not only exhibited pro-survival and pro-angiogenic activities, but also promoted proliferation of neonatal cardiomyocytes. These extracellular vesicles contain a cargo of proteins, mRNA and primary microRNA precursors that are enriched in exosomes and are capable of modulating collectively many of the cellular pathways involved in protein metabolism, cell growth, as well as cellular responses to stress and organisation of the extracellular matrix. Thus the W8B2+ CSC secretome contains a multitude of bioactive paracrine factors we have now characterised, that might well be harnessed for therapeutic application for cardiac repair and regeneration.

The benefits of adult stem cells for repair of the heart have been attributed to the repertoire of salutary paracrine activities they appear to exert. We previously isolated human W8B2+ cardiac stem cells (CSCs) and found they powerfully influence cardiomyocytes and endothelial cells to collectively promote cardiac repair and regeneration. Here, the complexity of the W8B2+ CSC secretomes was characterised and examined in more detail. Using ion exchange chromatography to separate soluble proteins based on their net surface charge, the secreted factors responsible for the pro-survival activity of W8B2+ CSCs were found within the low and medium cation fractions. In addition to the soluble proteins, extracellular vesicles generated from W8B2+ CSCs not only exhibited pro-survival and pro-angiogenic activities, but also promoted proliferation of neonatal cardiomyocytes. These extracellular vesicles contain a cargo of proteins, mRNA and primary microRNA precursors that are enriched in exosomes and are capable of modulating collectively many of the cellular pathways involved in protein metabolism, cell growth, as well as cellular responses to stress and organisation of the extracellular matrix. Thus the W8B2+ CSC secretome contains a multitude of bioactive paracrine factors we have now characterised, that might well be harnessed for therapeutic application for cardiac repair and regeneration.