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Introduction: Truncal and head and neck melanomas have a greater propensity to drain to multiple lymph node basins (MLNB) compared to extremity melanomas, which drain predominantly to a single lymph node basin (SLNB). The objective of this study was to compare the lymphatic drainage patterns and characteristics of truncal and head and neck melanoma, to assess their clinical usefulness in areas of unpredictable drainage. Methods: A retrospective review of 143 patients with head and neck or truncal melanoma from 2014–2018 treated at a tertiary referral hospital in Melbourne, Australia. Patients scheduled for a wide local excision (WLE) and SeNBx, and those whose initial biopsy results stated melanoma type, cell type and BT, were included. Patients were excluded if they underwent nodal biopsy alone. Results: We identified 95 patients with truncal and 48 with head and neck melanoma. Drainage to MLNB was significantly higher in the truncal melanoma group (36.8%) compared to the head and neck group (10.4%) (P = 0.001). Patients with drainage to MLNB had a higher positive sentinel node biopsy (SeNBx) rate compared to those with SLNB (40.0% verus. 12.6%, P

Background: The occurrence of radioulnar shaft fracture and elbow dislocation simultaneously is not a usual event, and it has only been reported in a few studies. This study aimed to report a case with posterior ulno-humeral joint dislocation and concomitant radioulnar shaft fracture. Furthermore, in a literature review, we summarized other studies that reported this kind of fracturedislocation and the treatment chosen in each study. Case Report: A 12-year-old boy was admitted to the emergency department due to high-energy trauma following a car accident while cycling. Open fracture of the left radioulnar shaft and posterior dislocation of the left elbow were detected. Conclusion: More studies are required to advance our knowledge about this kind of fracture-dislocation, especially its mechanism of injury. Subsequent examinations can help us design an efficient classification system and determine the best treatment option for radioulnar shaft fracture with elbow dislocation.

Background: Complex regional pain syndrome (CRPS) is an important common complication after surgical treatment for distal radius fracture. Recognition of the related factors is important to reduce the burden of the problem. Herein, the contributing factors for CRPS after distal radius fracture surgery are studied. Methods: In this cross-sectional comparative study, 250 patients admitted to the orthopedics ward in Sina Hospital in Tehran, Iran, from 2017 to 2020, were enrolled. The contributing factors for CRPS after distal radius fracture surgery were determined in them. Results: The CRPS was seen in 17 cases (6.8%). 9 percent of male and 1.9 percent of female patients were diagnosed with CRPS. This showed a significant difference in the Fisher test (P = 0.028). The mean pain severity by Visual Analogue Scale (VAS) was higher in CRPS cases, that showed a significant difference by the Mann-Whitney test (P = 0.001). Conclusion: According to the obtained results, it may be concluded that CRPS was seen in seven percent of patients under surgery for distal radius fracture and was related to the male gender and higher pain severity.

Introduction: Surgery and radiotherapy are key cancer treatments and the leading causes of damage to the lymphatics, a vascular network critical to fluid homeostasis and immunity. The clinical manifestation of this damage constitutes a devastating side-effect of cancer treatment, known as lymphoedema. Lymphoedema is a chronic condition evolving from the accumulation of interstitial fluid due to impaired drainage via the lymphatics and is recognised to contribute significant morbidity to patients who survive their cancer. Nevertheless, the molecular mechanisms underlying the damage inflicted on lymphatic vessels, and particularly the lymphatic endothelial cells (LEC) that constitute them, by these treatment modalities, remain poorly understood.Methods: We used a combination of cell based assays, biochemistry and animal models of lymphatic injury to examine the molecular mechanisms behind LEC injury and the subsequent effects on lymphatic vessels, particularly the role of the VEGF-C/VEGF-D/VEGFR-3 lymphangiogenic signalling pathway, in lymphatic injury underpinning the development of lymphoedema.Results: We demonstrate that radiotherapy selectively impairs key LEC functions needed for new lymphatic vessel growth (lymphangiogenesis). This effect is mediated by attenuation of VEGFR-3 signalling and downstream signalling cascades. VEGFR-3 protein levels were downregulated in LEC that were exposed to radiation, and LEC were therefore selectively less responsive to VEGF-C and VEGF-D. These findings were validated in our animal models of radiation and surgical injury.Discussion: Our data provide mechanistic insight into injury sustained by LEC and lymphatics during surgical and radiotherapy cancer treatments and underscore the need for alternative non-VEGF-C/VEGFR-3-based therapies to treat lymphoedema.

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

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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.

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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: Recurrent pressure sores and unstable scars over the bony prominences of the greater trochanter and ischium are a troublesome sequela of spinal injury. A reliable reconstruction is needed for patients with ‘dual-defect’ pressure sores in these locations. We modified the pedicled anterolateral thigh (ALT) flap to fit the reconstructive requirements of ‘dual-defect’ pressure sores.Methods: Eleven consecutive patients with concurrent pressure sores (> grade III) or unstable scar in one of the ‘dual-defect’ areas and an active pressure sore in the other were identified from the Victorian state tertiary referral centre for spinal injuries. We describe the technique and clinical experience of pedicled ALT flaps for reconstruction of ‘dual-defect’ pressure sores in this patient cohort. Preoperative status and minor and major postoperative complications were recorded. Results: Eleven consecutive pedicled myocutaneous ALT flaps were performed for reconstruction of ‘dual-defect’ pressure sores. Several key variations in the anatomical landmarks and the intra-operative flap raise technique that are integral to the use of the ALT flap for this application are described herein. The average dimensions of the cutaneous pressure sore defects were 6 x 4.9 cm (greater trochanter) and 8.2 x 6.7 cm (ischial). The average dimensions of the cutaneous paddle of the flaps raised were 27.3 x 8.4 cm. Two postoperative complications necessitated return to theatre but no incidences of flap loss were recorded. Conclusions: The modified pedicled myocutaneous ALT provides a robust reconstructive solution for resurfacing ‘dual-defect’ pressure sores in spinal patients. Further recommendations for future technical adaptations are made.

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.

Background: Interlocked intramedullary nailing is the most common treatment for closed tibial fractures. Reaming is a fundamental step in this surgical technique, and reamer breakage is a rare yet challenging complication during this operation. Case Report: A 34-year old male with a tibial shaft fracture was admitted for early closed tibial nailing. During the reaming process, the reamer broke and stuck in the medulla at the isthmus level. We extracted the broken piece by back hammering a cannulated T-handle placed on the ball tip guide pin. Conclusion: In this closed and quick method, we did not use any extra device other than standard equipment of intramedullary nailing.

Background: Lentigo maligna (LM) characteristically has an ill-defined margin and may require multiple excisions to achieve complete excision with 5mm margins. In vivo reflectance confocal microscopy (RCM) is a non-invasive tool recognised as useful in the management of LM. The authors aimed to determine whether the use of RCM prior to surgical excision reliably increased the rate of complete excision when compared with standard surgical excision. Methods: This prospective pilot study included patients with biopsy-proven LM of head and neck region who sought consultation for surgical management from May 2017 to May 2019 at the Victorian Melanoma Service, Melbourne. Patients were randomised to two groups based on the availability of RCM — Group 1, RCM-guided surgical excision, and Group 2, standard surgical excision. Outcomes were measured based on clinical markings and histopathological margins achieved and reported as RCM or surgical margin excess or deficit. Ethics approval for the study was obtained from the Research Governance Unit of The Alfred Hospital, Melbourne. Results: Results demonstrated that RCM marking of lesion margins was in excess in 69 percent of cases, in deficit in 22 per cent and accurate in nine per cent after histopathological analysis of the specimens. In comparison, lesions that were surgically marked were removed with margins in excess in 43 per cent of cases, in deficit in 11 per cent and accurate in 46 per cent. Conclusion: This pilot study demonstrates that RCM did not increase the accuracy of LM surgical excision in comparison with standard surgically marked excisions.

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.

Background There is a general consensus among reconstructive surgeons that preoperative radiotherapy is associated with a higher risk of flap failure and complications in head and neck surgery. Opinion is also divided regarding the effects of radiation dose on free flap outcomes and timing of preoperative radiation to minimize adverse outcomes. Our meta-analysis will attempt to address these issues. Method A systematic review of the literature was conducted in concordance to PRISMA protocol. Data were combined using STATA 12 and Open Meta-Analyst software programmes. Results Twenty-four studies were included comparing 2842 flaps performed in irradiated fields and 3491 flaps performed in non-irradiated fields. Meta-analysis yielded statistically significant risk ratios for flap failure (RR 1.48, P = 0.004), complications (RR 1.84, P 60 Gy radiation had a non-statistically significant higher risk of flap failure (RR 1.61, P = 0.145). Conclusion Preoperative radiation is associated with a statistically significant increased risk of flap complications, failure and fistula. Preoperative radiation in excess of 60 Gy after radiotherapy represents a potential risk factor for increased flap loss and should be avoided where possible.

Summary Introduction The supraclavicular flap has re-emerged as a feasible option for head and neck reconstruction where a thin, pliable donor tissue is required or where free flap techniques may impose too great a surgical risk in frail patients. Whilst our understanding of the vasculature of this flap has improved immensely, the microvasculature and in particular the venous drainage of the distal half of the supraclavicular flap remain relatively unclear. The present study aims to detail the arterial supply and venous drainage of the supraclavicular flap, particularly relating to the interperforator anastomoses. Methods The arterial and venous systems of the supraclavicular flap were injected with a radiopaque medium in eighteen fresh cadavers (twenty three and twelve flaps, respectively). Dissected supraclavicular flaps were subjected to plain X-ray imaging plus CT angiography to visualise the arterial and venous systems and relations to surrounding muscle and fascia. Further, the nature of any true or choke anastomoses was described. Results The arterial supply of the supraclavicular flap is a combination of axial pattern proximally and random pattern distally, demarcated by the origin of deltoid muscle. The venous system must undergo significant shunting into high pressure vessels once the preferred natural venous pathways are ligated whilst the flap is raised. A vast number of anastomoses, both arterial and venous exist over the body of deltoid and are critical to distal flap tissue survival. Conclusion The vascularity of the supraclavicular flap is complex and relies upon relatively small, superficial vessels. This technique is a prototypical example of the angiosome concept at work and thus relies heavily on our understanding of the location and nature of the anastomoses along its course. We conclude that the supraclavicular flap remains a reliable method for reconstructing head and neck defects so long as the constraints of a complicated suprafascial vascular system are respected.

Background There are minimal data in the literature regarding the lymphatic drainage of the conjunctiva and lower eyelid and the relationship with postoperative chemosis and edema. Methods Injection, microdissection, and histologic and radiologic studies were conducted on 12 hemifacial fresh cadaver specimens. Indocyanine green lymphography was conducted in five volunteers. Results Histology identified lymphatic vessels superficial and deep to the orbicularis oculi. Cadaveric dissection, injection, and radiographic studies identified interconnecting superficial and deep facial lymphatic systems and a conjunctival lymphatic network draining through the tarsal plate to the deep lymphatic system. The superficial lymphatic collectors traveled in subcutaneous fat within the lateral orbital and nasolabial fat compartments. The lateral deep lymphatic collectors traveled beneath orbicularis oculi, then through the superficial orbicularis retaining ligament, and into the sub-orbicularis oculi fat in the roof of the prezygomatic space. These vessels descended to preperiosteal fat at the level of zygomaticocutaneous ligaments to travel adjacent to the facial nerve into preauricular nodes. Indocyanine green lymphography identified correlating draining pathways laterally to the parotid nodes and medially to submandibular nodes. Conclusions The authors have found that the lower eyelid and conjunctiva are drained by interconnecting superficial and deep lymphatic systems of the face. The superficial system is vulnerable to damage in incisions and dissection in the infraorbital area. The deep system is vulnerable to damage in dissection around the orbicularis retaining ligament and the zygomaticocutaneous ligaments. The authors suggest that concurrent damage to both the superficial and deep lymphatic systems, especially laterally, may be responsible for postoperative chemosis and edema.

Reconstructive requirements of medium to large sized oral mucosal defects following oncological resection include restoration of mucosal continuity with prevention of salivary leak and fistula formation, predictable soft tissue healing, and ensuring optimal oral function and cosmetic restoration. Such defects frequently mandate the use of microvascular free tissue transfer of fasciocutaneous flaps such as the radial forearm or anterolateral thigh flaps, or, for larger defects incorporating significant dead-space, muscle flaps such as rectus abdominis or latissimus dorsi. Commonly described techniques for re-establishing continuity of the epithelial component include using native mucosa, split skin graft, or a myocutaneous flap skin paddle. Few case series reports exist of non-epithelial reconstructive approaches. Here, the authors report a large series of muscle only flaps for oral defect reconstruction following oncologic resection. The current study demonstrates that mucosalised muscle is an effective additional method for intra-oral mucosal defect reconstruction.

Tumor metastasis to lymph nodes is a key indicator of patient survival, and is enhanced by the neo-lymphatics induced by tumor-secreted VEGF-C or VEGF-D, acting via VEGFR-3 signalling. These targets constitute important avenues for anti-metastatic treatment. Despite this new understanding, clinical observations linking metastasis with tumor depth or location suggest that lymphangiogenic growth factors are not the sole determinants of metastasis. Here we explored the influence of tumor proximity to lymphatics capable of responding to growth factors on nodal metastasis in a murine VEGF-D over-expression tumor model. We found that primary tumor location profoundly influenced VEGF-D-mediated lymph node metastasis: 89 % of tumors associated with the flank skin metastasised, in contrast with only 19 % of tumors located more deeply on the body wall (p

Shayan R, Karnezis T, Murali R, Wilmott J S, Ashton M W, Taylor G I, Thompson J F, Hersey P, Achen M G, Scolyer R A & Stacker S A (2012) Histopathology 61, 702–710 Lymphatic vessel density in primary melanomas predicts sentinel lymph node status and risk of metastasis Aims: Important prognostic factors in patients with cutaneous melanoma include primary tumour thickness/depth of invasion, ulceration and mitotic rate, and the presence of tumour cells in regional lymph nodes. More recently, features of stromal components, such as blood and lymphatic vessel density, have been suggested as additional indicators of metastatic potential. Our aim was to investigate the relationship between tumour lymphatic vessels and lymph node metastasis. Methods and results: Metastasizing (n = 11) and non-metastasizing (n = 11) primary melanoma samples matched for depth/thickness, mitotic rate and ulceration were examined for lymphatic vessel density (LVD) in the primary tumour, using an antibody to podoplanin. Significant differences were found between LVD (vessels/unit area) in the peripheral (5.73 ± 0.67) versus central (1.72 ± 0.42) regions of the metastasizing tumour group (P

SummaryLymphatic metastasis is facilitated by lymphangiogenic growth factors VEGF-C and VEGF-D that are secreted by some primary tumors. We identified regulation of PGDH, the key enzyme in prostaglandin catabolism, in endothelial cells of collecting lymphatics, as a key molecular change during VEGF-D-driven tumor spread. The VEGF-D-dependent regulation of the prostaglandin pathway was supported by the finding that collecting lymphatic vessel dilation and subsequent metastasis were affected by nonsteroidal anti-inflammatory drugs (NSAIDs), known inhibitors of prostaglandin synthesis. Our data suggest a control point for cancer metastasis within the collecting lymphatic endothelium, which links VEGF-D/VEGFR-2/VEGFR-3 and the prostaglandin pathways. Collecting lymphatics therefore play an active and important role in metastasis and may provide a therapeutic target to restrict tumor spread.

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.

Glioblastoma multiforme is the most aggressive and lethal tumor of the central nervous system with limited treatment strategies on offer, and as such the identification of effective novel therapeutic agents is paramount. To examine the efficacy of proteasome inhibitors, we tested bortezomib, carfilzomib, nafamostat mesylate, gabexate mesylate and acetylsalicylic acid on glioblastoma cell viability, migration and invasion. Both bortezomib and carfilzomib produced significant reduction of cell viability, while nafamostat mesylate, gabexate mesylate and acetylsalicylic acid did not. Subsequent testing showed that carfilzomib significantly reduced cell viability at nM concentrations. Carfilzomib also reduced cell migration, secretion and activation of MMP2 and also cell invasion of all four glioblastoma cells tested. In summary, carfilzomib represents a novel, yet FDA-approved agent for the treatment of glioblastoma multiforme.

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.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.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%).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.

OBJECTIVES: The incidence of animal bite injuries in Australia is high. There is currently no established method for reliably predicting whether a patient with a bite injury will require admission to hospital or surgery. DESIGN: A retrospective audit of mammalian bite injuries at seven major hospitals in Melbourne, Victoria, over a 2-year period. The associations between each predictor and outcome of interest were analysed with univariate and multiple regression analyses. SETTING: Seven major hospitals in Melbourne, Victoria: the Alfred Hospital, Austin Hospital, Frankston Hospital, Monash Medical Centre, Royal Melbourne Hospital, St Vincent's Hospital and Western Hospital. PARTICIPANTS: Patients presenting to emergency departments with mammalian bite injuries. MAIN OUTCOME MEASURES: Hospital admission, intravenous antibiotic therapy, surgery, reoperation, readmission. RESULTS: We identified 717 mammalian bite injuries. The mean age of the patients was 36.5 years (median, 34 years; range, 0-88 years), with an equal number of males and females. The overall rate of hospital admission was 50.8%, and the mean length of stay was 2.7 days. Intravenous antibiotics were administered in 46% of cases; surgery was undertaken in 43.1% of cases. The re-operation rate was 4.5%, the re-admission rate was 3%. CONCLUSIONS: Our study provides a detailed epidemiological analysis of animal bite injuries at seven major hospitals in Victoria. Risk factors for hospitalisation and surgery may assist in identifying patients who require admission and surgical intervention. Language: en

Distant organ metastasis is the most important factor in determining patient survival in cancer. This is thought to occur via the body's own systems for transporting fluid and cells, the blood vascular and lymphatic systems. Cancer cells may exploit these vascular systems by expressing growth factors, which alter the normal pattern of angiogenesis and lymphatic vessel growth (lymphangiogenesis), thus creating conduits for tumour metastasis. With respect to lymphatic metastasis, techniques which allow the mapping of a tumour's lymphatic drainage and sampling of the 'sentinel node' from the regional lymph node group provide crucial prognostic information, determine further treatment and offer a window into tumour-host immune interactions. Aberrant drainage patterns so identified are both clinically significant, and highlight important anatomical and molecular complexities not explained by existing models of lymphatic development or anatomy. The molecular controls of tumour lymphangiogenesis and factors determining which lymphatic vessel subtypes are induced may be targets for novel therapeutics designed to restrict cancer metastasis. Furthermore, analyses of these control mechanisms will enhance our understanding of the interactions between the tumour cells and the lymphatic vasculature. For many years, disparate groups of clinical researchers and basic scientists have been working to unravel the mysteries of the lymphatic system. This review aims to summarize these contributions, in terms of the history, identification, structure and function of lymphatic vessels in cancer and the role they play in tumour metastasis. Current ideas about the roles of lymphangiogenic growth factors, their signalling pathways in lymphatic metastasis and therapeutic opportunities to restrict this spread will also be explored.

There is a general consensus among reconstructive surgeons that preoperative radiotherapy is associated with a higher risk of flap failure and complications in head and neck surgery. Opinion is also divided regarding the effects of radiation dose on free flap outcomes and timing of preoperative radiation to minimize adverse outcomes. Our meta-analysis will attempt to address these issues.A systematic review of the literature was conducted in concordance to PRISMA protocol. Data were combined using STATA 12 and Open Meta-Analyst software programmes.Twenty-four studies were included comparing 2842 flaps performed in irradiated fields and 3491 flaps performed in non-irradiated fields. Meta-analysis yielded statistically significant risk ratios for flap failure (RR 1.48, P = 0.004), complications (RR 1.84, P0.001), reoperation (RR 2.06, P0.001) and fistula (RR 2.05, P0.001). Mean radiation dose demonstrated a trend towards increased risk of flap failure, but this was not statistically significant. On subgroup analysis, flaps with60 Gy radiation had a non-statistically significant higher risk of flap failure (RR 1.61, P = 0.145).Preoperative radiation is associated with a statistically significant increased risk of flap complications, failure and fistula. Preoperative radiation in excess of 60 Gy after radiotherapy represents a potential risk factor for increased flap loss and should be avoided where possible.

The generation of new lymphatic vessels through lymphangiogenesis and the remodelling of existing lymphatics are thought to be important steps in cancer metastasis. The past decade has been exciting in terms of research into the molecular and cellular biology of lymphatic vessels in cancer, and it has been shown that the molecular control of tumour lymphangiogenesis has similarities to that of tumour angiogenesis. Nevertheless, there are significant mechanistic differences between these biological processes. We are now developing a greater understanding of the specific roles of distinct lymphatic vessel subtypes in cancer, and this provides opportunities to improve diagnostic and therapeutic approaches that aim to restrict the progression of cancer.

Over the past decade, research investigating the development of the lymphatic vascular system has become a key focus within the fields of developmental biology and cancer biology. Critical discoveries relating to the specification of lymphatic endothelial cells and lymphatic morphogenesis during embryonic development have helped to identify novel molecular drivers of pathological lymphangiogenesis; that is, the sprouting of a new lymphatic vessel from, or the enlargement of, pre-existing lymphatic vessels. These pathways may also constitute potentially useful therapeutic targets, which may be exploited to restrict tumor spread via the lymphatic system. Here, we discuss the current knowledge of both developmental and tumor-induced lymphangiogenesis, and draw parallels between the two processes to describe the molecular pathways that are re-capitulated during the growth of tumor lymphatics, and which promote metastasis.