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Beaches are major sites of microbiological pollution. Assessment of the abundance of resistant forms of enteric protozoa on these recreational waters is important for the prevention and management of health risks. Based on sedimentation and flotation methods, this study found that Kribi beach waters concentrate considerable amounts of enteric protozoa, which are potentially pathogenic. They include Coccidia (Cryptosporidium sp. and Cyclospora cayetanensis), Amoebae (Endolimax nana, Entamoeba histolytica and Entamoeba coli) and Flagellates (Giardia intestinalis). In general, seasonal changes and tidal cycles have significantly impacted the spread of these parasites along Kribi beaches. Thus, at all sites surveyed (Mpalla, Ngoyè and Mboamanga), maximum protozoan abundances were recorded at low tide and during the rainy seasons. It should also be noted that at each sampling site, significant correlations were recorded between certain protozoa and certain physico-chemical variables (p < 0.05). At Mboamanga, for example, Cryptosporidium sp. and Endolimax nana were positively correlated during the Short Rainy Season with temperature (r = 0.601, p = 0.044 and r = 0.632, p = 0.042). At Mpalla, a positive and significant correlation was observed during the Short Rainy Season between Entamoeba coli and pH (r = 0.605, p = 0.033). The high concentration of resistant forms of these enteric protozoa at Kribi beaches is a real public health threat for bathers. Therefore, in this tourist town, it is urgent to put in place an effective plan for the collection and sustainable treatment of solid and liquid waste, which are the main sources of contamination.

SCLC is a highly aggressive tumor with a 5-year survival rate of less than 6%. A heterogeneous disease, SCLC is classified into four subtypes that include tumors with neuroendocrine and non-neuroendocrine features. Immune checkpoint blockade has been recently added for the frontline treatment of SCLC; however, this therapy has only led to modest clinical improvements. The lack of clinical benefit in a cancer type known to have a high tumor mutational burden has been attributed to poor T-cell infiltration and low expression of MHC-class I in most SCLC tumors. In an attempt to devise a more effective immunotherapeutic regimen, this study investigated an alternate approach on the basis of the use of the clinical-stage interleukin-15 superagonist, N-803.Preclinical models of SCLC spanning all molecular subtypes were used to evaluate the susceptibility of SCLC to natural killer (NK)-mediated lysis in vitro, including NK cells activated by N-803. Antitumor activity of N-803 was evaluated in vivo with a xenograft model of SCLC.In vitro and in vivo data revealed differences in susceptibility of SCLC subtypes to lysis by NK cells and that NK cells activated by N-803 effectively lyse SCLC tumor cells across all variant subtypes, regardless of their expression of MHC-class I.These findings highlight the potential of a novel immune-based intervention using a cytokine-based therapeutic option for the treatment of SCLC. We hypothesize that N-803 may provide benefit to most patients with SCLC, including those with immunologically cold tumors lacking MHC expression.

To evaluate the feasibility and early outcomes of below the ankle orbital atherectomy in chronic limb-threatening ischemia in patients with calcified foot arteries.12 patients (mean age 69.4 ± 14.7; range 57 to 85 years) who were affected by diabetes underwent orbital atherectomy below the knee and ankle arterial segments. Technical success was defined as orbital atherectomy passage and debulked the calcified lesion, delivery of adjunctive therapy, and30% residual stenosis at final angiogram. The procedural outcome included complications, amputation-free survival, and freedom from clinically-driven target lesion revascularization at 30-days and 6-months of follow-up.Orbital atherectomy was performed in 3 cases in Anterior tibial (AT) and dorsalis pedis (Ped) arteries + Posterior tibial (PT) and Lateral Plantar (Lat Plan), 5 cases in PT and Lat Plan arteries, 1 case of PT and Medial Plantar, 1 case of Peroneal and Plan Lat, and 2 cases of AT and Ped. After atherectomy, we used a drug-coated balloon (DCB) angioplasty. Technical success was achieved in 11 (91.6%) cases. No deaths were registered during the follow-up. The limb salvage rate was 100%, and no major amputations were registered. Amputation-free survival was 50%. Freedom from CD-TLR was 100% at 30 days and 91.7% at 6-months. One patient underwent a TLR at three months. No major cardiovascular events, limb events, or significant procedure-related complications were registered.CSI orbital atherectomy and DCB angioplasty appear a feasible and promissor treatment options in diabetic CLTI patients.

This large, international dataset contains survey responses from N = 12,570 students from 100 universities in 35 countries, collected in 21 languages. We measured anxieties (statistics, mathematics, test, trait, social interaction, performance, creativity, intolerance of uncertainty, and fear of negative evaluation), self-efficacy, persistence, and the cognitive reflection test, and collected demographics, previous mathematics grades, self-reported and official statistics grades, and statistics module details. Data reuse potential is broad, including testing links between anxieties and statistics/mathematics education factors, and examining instruments’ psychometric properties across different languages and contexts. Data and metadata are stored on the Open Science Framework website [https://osf.io/mhg94/].

Therapeutic advances in the treatment of acute lymphoblastic leukemia (ALL) have increased the number of survivors but have promoted the development of long-term side effects, the best documented of which is obesity. The present retrospective case series analyzed data collected at diagnosis, end of treatment, and last follow-up visit of 210 ALL survivors treated between August 2005 and October 2014. Clinical and anthropometric data were collected from medical records. The nutritional diagnosis was based on

Deception research has shown that analysing verbal content can be effective to distinguish between truths and lies. However, most verbal cues are cues to truthfulness (truth tellers report the cue more than lie tellers), whereas cues to deception (lie tellers report the cue more than truth tellers) are largely absent. The complication approach, measuring complications (cue to truthfulness), common knowledge details (cue to deception), self-handicapping strategies (cue to deception), and the ratio of complications, aims to fill this gap in the literature. The present experiment examined the effectiveness of the complication approach when varying the amount of lying, with an Italian sample. Seventy-eight participants were assigned to one of three different experimental conditions: Truth tellers (telling the truth about the event), embedders (providing a mixture of truthful and false information) and outright lie tellers (providing false information). Participants were interviewed about a past experience concerning an out of the ordinary event. Complications discriminated truth tellers from lie tellers. The absence of significant effects for common knowledge details and self-handicapping strategies, the limitations of the experiment and suggestions for future research are discussed.

Supplementary figures and tables

Progressive tumor growth is associated with deficits in the immunity generated against tumor antigens. Vaccines targeting tumor neoepitopes have the potential to address qualitative defects; however, additional mechanisms of immune failure may underlie tumor progression. In such cases, patients would benefit from additional immune-oncology agents targeting potential mechanisms of immune failure. This study explores the identification of neoepitopes in the MC38 colon carcinoma model by comparison of tumor to normal DNA and tumor RNA sequencing technology, as well as neoepitope delivery by both peptide- and adenovirus-based vaccination strategies. To improve antitumor efficacies, we combined the vaccine with a group of rationally selected immune-oncology agents. We utilized an IL15 superagonist to enhance the development of antigen-specific immunity initiated by the neoepitope vaccine, PD-L1 blockade to reduce tumor immunosuppression, and a tumor-targeted IL12 molecule to facilitate T-cell function within the tumor microenvironment. Analysis of tumor-infiltrating leukocytes demonstrated this multifaceted treatment regimen was required to promote the influx of CD8+ T cells and enhance the expression of transcripts relating to T-cell activation/effector function. Tumor-targeted IL12 resulted in a marked increase in clonality of T-cell repertoire infiltrating the tumor, which when sculpted with the addition of either a peptide or adenoviral neoepitope vaccine promoted efficient tumor clearance. In addition, the neoepitope vaccine induced the spread of immunity to neoepitopes expressed by the tumor but not contained within the vaccine. These results demonstrate the importance of combining neoepitope-targeting vaccines with a multifaceted treatment regimen to generate effective antitumor immunity.

PDF file - 89K, TGF-β1 secretion in LNCap and H520 cells.

PDF file - 326K, Treatment with SD-208 reverts the mesenchymal phenotype of colon carcinoma cell lines.

Purpose: The conversion of tumor cells from an epithelial to a mesenchymal-like phenotype, via a process designated as the epithelial–mesenchymal transition (EMT), is known to mediate tumor resistance to a variety of cell death inducers, including cytotoxic effector immune cells. The goal of this study was to identify and potentially repurpose FDA-approved compounds capable of reducing mesenchymal features of human lung carcinoma cells, which could be used in combination with immunotherapies or chemotherapeutic strategies to improve clinical responses.Experimental Design: In the current report, we have utilized a quantitative high-throughput screening (qHTS) of a pharmaceutical collection of more than 2,000 compounds to identify clinically approved drugs capable of augmenting the sensitivity of mesenchymal-like, lung cancer cells to immune- and chemotherapy-mediated lysis, both in vitro and in vivo.Results: The estrogen receptor antagonist fulvestrant was shown to reduce mesenchymal features of lung carcinoma cells, resulting in tumor sensitization to the cytotoxic effect of antigen-specific T cells, natural killer (NK) effector cells, and chemotherapy both in vivo and in vitro.Conclusions: To our knowledge, this is the first report defining a potential role for estrogenic signaling in promoting tumor resistance to immune-mediated cytotoxicity and chemotherapy in lung cancer. Our data demonstrate a robust association between the acquisition of mesenchymal attributes, therapeutic resistance of lung carcinoma cells, and the expression of estrogen receptor 1 (ESR1), supporting further investigations on the role of estrogen signaling in lung cancer progression via the induction of EMT. Clin Cancer Res; 22(24); 6204–16. ©2016 AACR.

Supplemental Figure S7. SX-682 treatment does not alter PMN-MDSC suppressive capacity

Supplementary Data from Pilot Study of Vaccination with Recombinant CEA-MUC-1-TRICOM Poxviral-Based Vaccines in Patients with Metastatic Carcinoma

Purpose: The epithelial–mesenchymal transition (EMT) is emerging as a critical factor for the progression and metastasis of carcinomas, as well as drug resistance. The T-box transcription factor Brachyury has been recently characterized as a driver of EMT in human carcinoma cells. The purpose of this study was to characterize Brachyury as a potential target for lung cancer therapy.Experimental Design: The expression of Brachyury was evaluated by PCR and by immunohistochemistry in human lung tumors and adult normal tissues. Brachyury gene copy number and promoter methylation status were analyzed in tumor tissues with various levels of Brachyury expression. Lung carcinoma cells' susceptibility to T-cell lysis and EGF receptor (EGFR) kinase inhibition were also evaluated relative to the levels of Brachyury.Results: Our results showed Brachyury protein expression in 41% of primary lung carcinomas, including 48% of adenocarcinomas and 25% of squamous cell carcinomas. With the exception of normal testis and some thyroid tissues, the majority of normal tissues evaluated in this study were negative for the expression of Brachyury protein. Brachyury-specific T cells could lyse Brachyury-positive tumors and the level of Brachyury corresponded to resistance of tumor cells to EGFR kinase inhibition.Conclusion: We hypothesize that the elimination of Brachyury-positive tumor cells may be able to prevent and/or diminish tumor dissemination and the establishment of metastases. The ability of Brachyury-specific T-cell lines to lyse Brachyury-positive tumor cells, in vitro, supports the development of Brachyury-based immunotherapeutic approaches for the treatment of lung cancer. Clin Cancer Res; 18(14); 3868–79. ©2012 AACR.

PDF file, 62KB, Analysis of Brachyury gene copy number and promoter methylation status.

PDF file, 55KB, Number and percentage of tissues from adult non-cancer subjects that were positive for Brachyury expression by immunohistochemistry.

Purpose:BN-CV301 is a poxviral-based vaccine comprised of recombinant (rec.) modified vaccinia Ankara (MVA-BN-CV301; prime) and rec. fowlpox (FPV-CV301; boost). Like its predecessor PANVAC, BN-CV301 contains transgenes encoding tumor-associated antigens MUC1 and CEA as well as costimulatory molecules (B7.1, ICAM-1, and LFA-3). PANVAC was reengineered to make it safer and more antigenic.Patients and Methods:This open-label, 3+3 design, dose-escalation trial evaluated three dose levels (DL) of MVA-BN-CV301: one, two, or four subcutaneous injections of 4 × 108 infectious units (Inf.U)/0.5 mL on weeks 0 and 4. All patients received FPV-CV301 subcutaneously at 1 × 109 Inf.U/0.5 mL every 2 weeks for 4 doses, then every 4 weeks. Clinical and immune responses were evaluated.Results:There were no dose-limiting toxicities. Twelve patients enrolled on trial [dose level (DL) 1 = 3, DL2 = 3, DL3 = 6). Most side effects were seen with the prime doses and lessened with subsequent boosters. All treatment-related adverse events were temporary, self-limiting, grade 1/2, and included injection-site reactions and flu-like symptoms. Antigen-specific T cells to MUC1 and CEA, as well as to a cascade antigen, brachyury, were generated in most patients. Single-agent BN-CV301 produced a confirmed partial response (PR) in 1 patient and prolonged stable disease (SD) in multiple patients, most notably in KRAS-mutant gastrointestinal tumors. Furthermore, 2 patients with KRAS-mutant colorectal cancer had prolonged SD when treated with an anti-PD-L1 antibody following BN-CV301.Conclusions:The BN-CV301 vaccine can be safely administered to patients with advanced cancer. Further studies of the vaccine in combination with other agents are planned.See related commentary by Repáraz et al., p. 4871

Characterization of rF-brachyury-TRICOM

Detailed description of the high-throughput screen assay

Pixel analysis of immunohistochemical staining of various markers in tumor xenografts shown in Figure 6.

Supplementary Table S1A: Phenotypic analysis of surface markers on human DCs transfected with MVA-BN-CV301 and FPV-CV301 vectors Supplementary Table S1B: Activation of CEA- and MUC1-specific T-cells by MVA-BN-CV301 and FPV-CV301 Supplementary Table S2: Number of patients reporting adverse events possibly, probably or definitely attributed to CV301 (n=202 events in total) Supplementary Table S3: Development of CEA- and MUC1-specific T-cells by BN-CV301 Supplementary Figure S1: KRAS specific responses assessed in 3 patients preâ€' and postâ€'BN- CV301 vaccine

PDF file, 113KB, Patient characteristics for samples in human lung cancer cDNA arrays.

Supplemental Figure S4. Ex vivo SX-682 exposure does not reduce the viability of sorted MDSC

Supplemental Figure S2. PMN-MDSC can be isolated from tumor single cell suspensions

Purpose: The transcription factor brachyury has been shown in preclinical studies to be a driver of the epithelial-to-mesenchymal transition (EMT) and resistance to therapy of human tumor cells. This study describes the characterization of a Modified Vaccinia Ankara (MVA) vector–based vaccine expressing the transgenes for brachyury and three human costimulatory molecules (B7.1, ICAM-1, and LFA-3, designated TRICOM) and a phase I study with this vaccine.Experimental Design: Human dendritic cells (DC) were infected with MVA-brachyury-TRICOM to define their ability to activate brachyury-specific T cells. A dose-escalation phase I study (NCT02179515) was conducted in advanced cancer patients (n = 38) to define safety and to identify brachyury-specific T-cell responses.Results: MVA-brachyury-TRICOM-infected human DCs activated CD8+ and CD4+ T cells specific against the self-antigen brachyury in vitro. No dose-limiting toxicities were observed due to vaccine in cancer patients at any of the three dose levels. One transient grade 3 adverse event (AE) possibly related to vaccine (diarrhea) resolved without intervention and did not recur with subsequent vaccine. All other AEs related to vaccine were transient and ≤grade 2. Brachyury-specific T-cell responses were observed at all dose levels and in most patients.Conclusions: The MVA-brachyury-TRICOM vaccine directed against a transcription factor known to mediate EMT can be administered safely in patients with advanced cancer and can activate brachyury-specific T cells in vitro and in patients. Further studies of this vaccine in combination therapies are warranted and planned. Clin Cancer Res; 23(22); 6833–45. ©2017 AACR.

Supplemental Figure S8. SX-682 does not directly alter MOC2 cells in vitro

Purpose: Poxviral vectors have a proven safety record and can be used to incorporate multiple transgenes. Prior clinical trials with poxviral vaccines have shown that immunologic tolerance to self-antigens can be broken. Carcinoembryonic antigen (CEA) and MUC-1 are overexpressed in a substantial proportion of common solid carcinomas. The primary end point of this study was vaccine safety, with immunologic and clinical responses as secondary end points.Experimental Design: We report here a pilot study of 25 patients treated with a poxviral vaccine regimen consisting of the genes for CEA and MUC-1, along with a triad of costimulatory molecules (TRICOM; composed of B7.1, intercellular adhesion molecule 1, and lymphocyte function–associated antigen 3) engineered into vaccinia (PANVAC-V) as a prime vaccination and into fowlpox (PANVAC-F) as a booster vaccination.Results: The vaccine was well tolerated. Apart from injection-site reaction, no grade ≥2 toxicity was seen in more than 2% of the cycles. Immune responses to MUC-1 and/or CEA were seen following vaccination in 9 of 16 patients tested. A patient with clear cell ovarian cancer and symptomatic ascites had a durable (18-month) clinical response radiographically and biochemically, and one breast cancer patient had a confirmed decrease of >20% in the size of large liver metastasis.Conclusions: This vaccine strategy seems to be safe, is associated with both CD8 and CD4 immune responses, and has shown evidence of clinical activity. Further trials with this agent, either alone or in combination with immunopotentiating and other therapeutic agents, are warranted.

Purpose:Natural killer (NK)-cell–based immunotherapy may overcome obstacles to effective T-cell–based immunotherapy such as the presence of genomic alterations in IFN response genes and antigen presentation machinery. All immunotherapy approaches may be abrogated by the presence of an immunosuppressive tumor microenvironment present in many solid tumor types, including head and neck squamous cell carcinoma (HNSCC). Here, we studied the role of myeloid-derived suppressor cells (MDSC) in suppressing NK-cell function in HNSCC.Experimental Design:The ability of peripheral and tumor-infiltrating MDSC from mice bearing murine oral cancer 2 (MOC2) non-T-cell–inflamed tumors and from patients with HNSCC to suppress NK-cell function was studied with real-time impedance and ELISpot assays. The therapeutic efficacy of SX-682, a small-molecule inhibitor of CXCR1 and CXCR2, was assessed in combination with adoptively transferred NK cells.Results:Mice bearing MOC2 tumors pathologically accumulate peripheral CXCR2+ neutrophilic-MDSC (PMN-MDSC) that traffic into tumors and suppress NK-cell function through TGFβ and production of H2O2. Inhibition of MDSC trafficking with orally bioavailable SX-682 significantly abrogated tumor MDSC accumulation and enhanced the tumor infiltration, activation, and therapeutic efficacy of adoptively transferred murine NK cells. Patients with HNSCC harbor significant levels of circulating and tumor-infiltrating CXCR1/2+ CD15+ PMN-MDSC and CD14+ monocytic-MDSC. Tumor MDSC exhibited greater immunosuppression than those in circulation. HNSCC tumor MDSC immunosuppression was mediated by multiple, independent, cell-specific mechanisms including TGFβ and nitric oxide.Conclusions:The clinical study of CXCR1/2 inhibitors in combination with adoptively transferred NK cells is warranted.

Supplemental Figure S1. MOC2 tumors express myeloid chemokines

Methods not essential to the main conclusions but important for critical review of methods used for this work

Supplemental Figure S6. SX-682 treatment does not enhance MOC2 tumor T cell infiltration

Validation of immunohistochemistry staining method for estrogen receptor 1.

Numerous reports have now demonstrated that the epithelial-to-mesenchymal transition (EMT) process is involved in solid tumor progression, metastasis, and drug resistance. Several transcription factors have been implicated as drivers of EMT and metastatic progression, including Twist. Overexpression of Twist has been shown to be associated with poor prognosis and drug resistance for many carcinomas and other tumor types. The role of Twist in experimental cancer metastases has been principally studied in the 4T1 mammary tumor model, where silencing of Twist in vitro has been shown to greatly reduce in vivo metastatic spread. Transcription factors such as Twist are generally believed to be “undruggable” because of their nuclear location and lack of a specific groove for tight binding of a small molecule inhibitor. An alternative approach to drug therapy targeting transcription factors driving the metastatic process is T-cell–mediated immunotherapy. A therapeutic vaccine platform that has been previously characterized consists of heat-killed recombinant Saccharomyces cerevisiae (yeast) capable of expressing tumor-associated antigen protein. We report here the construction and characterization of a recombinant yeast expressing the entire Twist protein, which is capable of inducing both CD8+ and CD4+ Twist-specific T-cell responses in vivo. Vaccination of mice reduced the size of primary transplanted 4T1 tumors and had an even greater antitumor effect on lung metastases of the same mice, which was dependent on Twist-specific CD8+ T cells. These studies provide the rationale for vaccine-induced T-cell–mediated therapy of transcription factors involved in driving the metastatic process. Cancer Res; 74(7); 1945–57. ©2014 AACR.

PDF file - 92KB, Supplementary Table 1. Real-time PCR assays and PCR primers utilized.

Supplementary Figure 1 from IL-8 Signaling Plays a Critical Role in the Epithelial–Mesenchymal Transition of Human Carcinoma Cells

PDF file - 26KB, Figure S4. Resistance of brachyury-high tumor cells could be alleviated by WEE1 inhibition.

PDF file - 358KB, Expression of EMT marker expression in primary tumor and lung metastases.

PDF file - 138KB, Expression of murine Twist protein in S. cerevisiae Yeast.

PDF file - 122KB, Figure S2. Mechanism of resistance to immune-mediated cytotoxic lysis.

PDF file - 67KB, Figure S3. Brachyury affects CDK1 protein stability but not mRNA levels.

Aberrant expression of the T-box transcription factor brachyury in human carcinomas drives the phenomenon of epithelial–mesenchymal transition (EMT), a phenotypic modulation that facilitates tumor dissemination and resistance to conventional therapies, including chemotherapy and radiotherapy. By generating isogenic cancer cell lines with various levels of brachyury expression, we demonstrate that high levels of brachyury also significantly reduce the susceptibility of cancer cells to lysis by both antigen-specific T cells and natural killer cells. Our results indicated that resistance of brachyury-high tumor cells to immune-mediated attack was due to inefficient caspase-dependent apoptosis, manifested as inefficient nuclear lamin degradation in the presence of activated effector caspases. We correlated this phenomenon with loss of cell-cycle–dependent kinase 1 (CDK1), which mediates lamin phosphorylation. In support of a causal connection, pretreatment of tumor cells with a specific inhibitor of WEE1, a negative regulator kinase of CDK1, could counter the defective apoptosis of tumor cells expressing high levels of brachyury. Thus, our findings suggested that reconstituting CDK1 activity to threshold levels may be sufficient to restore immunosurveillance of mesenchymal-like cancer cells that have escaped previous immune detection or eradication. Cancer Res; 74(9); 2510–9. ©2014 AACR.

PDF file - 79KB, Schematic model of the proposed mechanism.

PDF file - 208KB, Recombinant Yeast-Twist vaccination does not induce toxicity.

O objetivo deste estudo foi avaliar o consumo alimentar e a qualidade de vida dos indivíduos com osteoartrose. Estudo do tipo série de casos, com pacientes adultos de ambos os sexos, onde foram avaliados dados socioeconômicos, antropométricos, dietéticos e sobre a qualidade de vida. Para avaliar o consumo alimentar foi utilizado um questionário de frequência alimentar do tipo qualitativo, com mensuração convertida em escores de frequência de consumo dos alimentos considerados de risco e de proteção para as doenças crônicas não transmissíveis. A qualidade de vida foi avaliada pelo questionário Short Form Health Survey Questionnaire - SF-12. Foram avaliados 70 indivíduos sendo 64,3% do sexo feminino, 44,3% com mais de 50 anos, bom nível de escolaridade em 55,7% com superior completo, 53,6% com excesso de peso e 33,3% apresentando outras comorbidades crônicas. Os escores de qualidade de vida foram abaixo de 50, o que podem indicar menor qualidade de vida. Observou-se que os alimentos mais consumidos por ambos os sexos foram aqueles considerados alimentos de risco. Com relação aos quartis dos escores de consumo alimentar, segundo características socioeconômicas e antropométricas, as pessoas que apresentavam outras doenças crônicas consumiam menos alimentos considerados protetores. E os alimentos considerados de risco foram mais consumidos pelo grupo com maior nível de renda. Quanto aos quartis dos escores de qualidade de vida segundo características socioeconômicas e antropométricas, nenhuma variável apresentou associação estatisticamente significante. A população avaliada apresenta riscos à sua saúde, devido à elevada frequência de excesso de peso e do baixo consumo de alimentos protetores, como frutas, legumes e verduras. Da mesma forma, mesmo não encontrando significância estatística, é possível observar uma tendência de piora na qualidade de vida de indivíduos portadores de OA.

Purpose, Retrograde recanalizations have gained increasing recognition in complex arterial occlusive disease. Re-entry devices are a well described adjunct for antegrade recanalizations. We present our experience with target balloon-assisted antegrade and retrograde recanalizations using re-entry devices in challenging chronic total occlusions. Materials and Methods: We report data from a retrospective multicenter registry. Eligibility criteria included either antegrade or retrograde use of the OutbackTM or GoBackTM re-entry catheter in combination with a balloon as a target to accomplish wire passage, when conventional antegrade and retrograde recanalization attempts had been unsuccessful. Procedural outcomes included technical success (defined as wire passage though the occlusion and delivery of adjunctive therapy with