Expanding the knowledge on lignocellulolytic and redox enzymes of worker and soldier castes from the lower termite coptotermes gestroi

Termites are considered one of the most efficient decomposers of lignocelluloses on Earth due to their ability to produce, along with its microbial symbionts, a repertoire of carbohydrate-active enzymes (CAZymes). Recently, a set of Pro-oxidant, Antioxidant, and Detoxification enzymes (PAD) were also correlated with the metabolism of carbohydrates and lignin in termites. The lower termite Coptotermes gestroi is considered the main urban pest in Brazil, causing damage to wood constructions. Recently, analysis of the enzymatic repertoire of C. gestroi unveiled the presence of different CAZymes. Because the gene profile of CAZy/PAD enzymes endogenously synthesized by C. gestroi and also by their symbiotic protists remains unclear, the aim of this study was to explore the eukaryotic repertoire of these enzymes in worker and soldier castes of C. gestroi. Our findings showed that worker and soldier castes present similar repertoires of CAZy/PAD enzymes, and also confirmed that endo-glucanases (GH9) and beta-glucosidases (GH1) were the most important glycoside hydrolase families related to lignocellulose degradation in both castes. Classical cellulases such as exo-glucanases (GH7) and endo-glucanases (GH5 and GH45), as well as classical xylanases (GH10 and GH11), were found in both castes only taxonomically related to protists, highlighting the importance of symbiosis in C. gestroi. Moreover, our analysis revealed the presence of Auxiliary Activity enzyme families (AAs), which could be related to lignin modifications in termite digestomes. In conclusion, this report expanded the knowledge on genes and proteins related to CAZy/PAD enzymes from worker and soldier castes of lower termites, revealing new potential enzyme candidates for second-generation biofuel processes7CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP140796/2013-4; 310186/2014-5; 442333/2014-511/20977-3; 15/06971-3; 12/19040-0; 14/10351-8; 06/59086-8; 14/20576- 7; 13/03061-0; 10/11469-1; 08/58037-9; 14/50371-8; 08/50114-

Oncoplastic Breast Consortium consensus conference on nipple-sparing mastectomy

Purpose Indications for nipple-sparing mastectomy (NSM) have broadened to include the risk reducing setting and locally advanced tumors, which resulted in a dramatic increase in the use of NSM. The Oncoplastic Breast Consortium consensus conference on NSM and immediate reconstruction was held to address a variety of questions in clinical practice and research based on published evidence and expert panel opinion. Methods The panel consisted of 44 breast surgeons from 14 countries across four continents with a background in gynecology, general or reconstructive surgery and a practice dedicated to breast cancer, as well as a patient advocate. Panelists presented evidence summaries relating to each topic for debate during the in-person consensus conference. The iterative process in question development, voting, and wording of the recommendations followed the modified Delphi methodology. Results Consensus recommendations were reached in 35, majority recommendations in 24, and no recommendations in the remaining 12 questions. The panel acknowledged the need for standardization of various aspects of NSM and immediate reconstruction. It endorsed several oncological contraindications to the preservation of the skin and nipple. Furthermore, it recommended inclusion of patients in prospective registries and routine assessment of patient-reported outcomes. Considerable heterogeneity in breast reconstruction practice became obvious during the conference. Conclusions In case of conflicting or missing evidence to guide treatment, the consensus conference revealed substantial disagreement in expert panel opinion, which, among others, supports the need for a randomized trial to evaluate the safest and most efficacious reconstruction techniques

Oncoplastic breast consortium recommendations for mastectomy and whole breast reconstruction in the setting of post-mastectomy radiation therapy

Aim: Demand for nipple-and skin-sparing mastectomy (NSM/SSM) with immediate breast reconstruction (BR) has increased at the same time as indications for post-mastectomy radiation therapy (PMRT) have broadened. The aim of the Oncoplastic Breast Consortium initiative was to address relevant questions arising with this clinically challenging scenario. Methods: A large global panel of oncologic, oncoplastic and reconstructive breast surgeons, patient advocates and radiation oncologists developed recommendations for clinical practice in an iterative process based on the principles of Delphi methodology. Results: The panel agreed that surgical technique for NSM/SSM should not be formally modified when PMRT is planned with preference for autologous over implant-based BR due to lower risk of long-term complications and support for immediate and delayed-immediate reconstructive approaches. Nevertheless, it was strongly believed that PMRT is not an absolute contraindication for implant-based or other types of BR, but no specific recom-mendations regarding implant positioning, use of mesh or timing were made due to absence of high-quality evidence. The panel endorsed use of patient-reported outcomes in clinical practice. It was acknowledged that the shape and size of reconstructed breasts can hinder radiotherapy planning and attention to details of PMRT techniques is important in determining aesthetic outcomes after immediate BR. Conclusions: The panel endorsed the need for prospective, ideally randomised phase III studies and for surgical and radiation oncology teams to work together for determination of optimal sequencing and techniques for PMRT for each patient in the context of BRPeer reviewe

Oncoplastic breast consortium recommendations for mastectomy and whole breast reconstruction in the setting of post-mastectomy radiation therapy

Aim Demand for nipple- and skin- sparing mastectomy (NSM/SSM) with immediate breast reconstruction (BR) has increased at the same time as indications for post-mastectomy radiation therapy (PMRT) have broadened. The aim of the Oncoplastic Breast Consortium initiative was to address relevant questions arising with this clinically challenging scenario. Methods A large global panel of oncologic, oncoplastic and reconstructive breast surgeons, patient advocates and radiation oncologists developed recommendations for clinical practice in an iterative process based on the principles of Delphi methodology. Results The panel agreed that surgical technique for NSM/SSM should not be formally modified when PMRT is planned with preference for autologous over implant-based BR due to lower risk of long-term complications and support for immediate and delayed-immediate reconstructive approaches. Nevertheless, it was strongly believed that PMRT is not an absolute contraindication for implant-based or other types of BR, but no specific recommendations regarding implant positioning, use of mesh or timing were made due to absence of high-quality evidence. The panel endorsed use of patient-reported outcomes in clinical practice. It was acknowledged that the shape and size of reconstructed breasts can hinder radiotherapy planning and attention to details of PMRT techniques is important in determining aesthetic outcomes after immediate BR. Conclusions The panel endorsed the need for prospective, ideally randomised phase III studies and for surgical and radiation oncology teams to work together for determination of optimal sequencing and techniques for PMRT for each patient in the context of BR

Oncoplastic Breast Consortium consensus conference on nipple-sparing mastectomy.

Purpose Indications for nipple-sparing mastectomy (NSM) have broadened to include the risk reducing setting and locally advanced tumors, which resulted in a dramatic increase in the use of NSM. The Oncoplastic Breast Consortium consensus conference on NSM and immediate reconstruction was held to address a variety of questions in clinical practice and research based on published evidence and expert panel opinion. Methods The panel consisted of 44 breast surgeons from 14 countries across four continents with a background in gynecology, general or reconstructive surgery and a practice dedicated to breast cancer, as well as a patient advocate. Panelists presented evidence summaries relating to each topic for debate during the in-person consensus conference. The iterative process in question development, voting, and wording of the recommendations followed the modified Delphi methodology. Results Consensus recommendations were reached in 35, majority recommendations in 24, and no recommendations in the remaining 12 questions. The panel acknowledged the need for standardization of various aspects of NSM and immediate reconstruction. It endorsed several oncological contraindications to the preservation of the skin and nipple. Furthermore, it recommended inclusion of patients in prospective registries and routine assessment of patient-reported outcomes. Considerable heterogeneity in breast reconstruction practice became obvious during the conference. Conclusions In case of conflicting or missing evidence to guide treatment, the consensus conference revealed substantial disagreement in expert panel opinion, which, among others, supports the need for a randomized trial to evaluate the safest and most efficacious reconstruction techniques

Report on a case of pure cribriform carcinoma of the breast with internal mammary node metastasis: description of the case and review of the literature

Aims and background: Pure ductal invasive carcinoma of the breast is more frequently associated with lymphatic invasion, lymph node involvement and high malignant histological grade than combined forms of breast carcinoma. Internal mammary node metastases are not frequently detected when the axillary nodes are negative. Patients and methods: We report our experience of a case of pure cribriform carcinoma, associated with an intraductal cribriform component, in which the sentinel axillary nodes were negative, while nodal and perinodal metastasis was detected in one internal mammary lymph node. Conclusions: Data from the literature show that positive internal mammary nodes are an unusual finding when the axillary sentinel node is negative, especially in cases of cancer with a relatively good prognosis, such as cribriform cases

Oncological Safety Of Autologous Fat Grafting After Breast Conservative Treatment: A Prospective Evaluation

Autologous fat graft to the breast is a useful tool to correct defects after breast conservative treatment (BCT). Although this procedure gains popularity, little is known about the interaction between the fat graft and the prior oncological environment. Evidences of safety of this procedure in healthy breast and after post-mastectomy reconstruction exist. However, there is paucity of data among patients who underwent BCT which are hypothetically under a higher risk of local recurrence (LR). Fifty-nine patients, with prior BCT, underwent 75 autologous fat graft procedures using the Coleman's technique, between October 2005 and July 2008. Follow-up was made by clinical and radiologic examination at least once, after 6 months of the procedure. Mean age was 50 ± 8.5 years, and mean follow-up was 34.4 ± 15.3 months. Mean time from oncological surgery to the first fat grafting procedure was 76.6 ± 30.9 months. Most of patients were at initial stage 0 (11.8%), I (33.8%), or IIA (23.7%). Immediate complication was observed in three cases (4%). Only three cases of true LR (4%) associated with the procedure were observed during the follow-up. Abnormal breast images were present in 20% of the postoperative mammograms, and in 8% of the cases, biopsy was warranted. Autologous fat graft is a safe procedure to correct breast defects after BCT, with low postoperative complications. Although it was not associated with increased risk of LR in the group of patients studied, prospective trials are needed to certify that it does not interfere in patient's oncological prognosis. © 2014 Wiley Periodicals, Inc.202159165Veronesi, U., Cascinelli, N., Mariani, L., Twenty-year follow-up of a randomized study comparing breast-conserving surgery with radical mastectomy for early breast cancer (2002) N Engl J Med, 347, pp. 1227-1232Fisher, B., Anderson, S., Bryant, J., Twenty-year follow-up of a randomized trial comparing total mastectomy, lumpectomy, and lumpectomy plus irradiation for the treatment of invasive breast cancer (2002) N Engl J Med, 347, pp. 1233-1241Veronesi, U., Saccozzi, R., Del Vecchio, M., Comparing radical mastectomy with quadrantectomy, axillary dissection, and radiotherapy in patients with small cancers of the breast (1981) N Engl J Med, 305, pp. 6-11Fisher, B., Bauer, M., Margolese, R., Five-year results of a randomized clinical trial comparing total mastectomy and segmental mastectomy with or without radiation in the treatment of breast cancer (1985) N Engl J Med, 312, pp. 665-673Wang, H.T., Barone, C.M., Steigelman, M.B., Aesthetic outcomes in breast conservation therapy (2008) Aesthet Surg J, 28, pp. 165-170Kotwall, C.A., Covington, D.L., Rutledge, R., Patient, hospital, and surgeon factors associated with breast conservation surgery: A statewide analysis in North Carolina (1996) Ann Surg, 224, pp. 419-429Parviz, M.A., Cassel, J.B., Kaplan, B.J., Breast conservation therapy rates are no different in medically indigent versus insured patients with early stage breast cancer (2004) J Surg Oncol, 84, pp. 57-62Kelemen III, J.J., Poulton, T., Surgical treatment of early-stage breast cancer in the Department of Defense healthcare system (2001) J Am Coll Surg, 192, pp. 293-297McGuire, K.P., Santillan, A.A., Kaur, P., Are mastectomies on the rise? A 13-year trend analysis of the selection of mastectomy versus breast conservation therapy in 5865 patients (2009) Ann Surg Oncol, 16, pp. 2682-2690Bajaj, A.K., Kon, P.S., Oberg, K.C., Miles, D.A.G., Aesthetic outcomes in patients undergoing breast conservation therapy for the treatment of localized breast cancer (2004) Plast Reconstr Surg, 14, pp. 1442-1449Clough, K.B., Thomas, S.S., Fitoussi, A.D., Reconstruction after conservative treatment for breast cancer: Cosmetic sequelae classification revisited (2004) Plast Reconstr Surg, 114, pp. 1743-1753Coleman, S.R., Saboeiro, A.P., Fat grafting to the breast revisited: Safety and efficacy (2007) Plast Reconstr Surg, 119, pp. 775-785Gosset, J., Flageul, G., Toussoun, G., Lipomodelling for correction of breast conservative treatment sequelae. Medicolegal aspects. Expert opinion on five problematic clinical cases (2008) Ann Chir Plast Esthet, 53, pp. 190-198Delay, E., Gosset, J., Toussoun, G., Efficacy of lipomodelling for the management of sequelae of breast cancer conservative treatment (2008) Ann Chir Plast Esthet, 53, pp. 153-168Missana, M.C., Laurent, I., Barreau, L., Autologous fat transfer in reconstructive breast surgery: Indications, technique and results (2007) EJSO, 33, pp. 685-690Gosset, J., Guerin, N., Toussoun, G., Radiological evaluation after lipomodelling for correction of breast conservative treatment sequelae (2008) Ann Chir Plast Esthet, 53, pp. 178-189Amar, O., Bruant-Rodier, C., Lehmann, S., Fat tissue transplant: Restoration of the mammary volume after conservative treatment of breast cancers, clinical and radiological considerations (2008) Ann Chir Plast Esthet, 53, pp. 169-177Gutowski, K.A., Current applications and safety of autologous fat grafts: A report of the ASPS fat graft task force (2009) Plast Reconstr Surg, 124, pp. 272-280. , ASPS Fat Graft Task ForceDelay, E., Sinna, R., Delaporte, T., Patient information before aesthetic lipomodeling (lipoaugmentation): A French plastic surgeon's perspective (2009) Aesthet Surg J, 29, pp. 386-395Schaffler, A., Scholmerich, J., Buechler, C., Mechanisms of disease: Adipokines and breast cancer - Endocrine and paracrine mechanisms that connect adiposity and breast cancer (2007) Nat Clin Pract Endocrinol Metab, 3, pp. 345-354Dieudonne, M.N., MacHinal-Quelin, F., Serazin-Leroy, V., Leptin mediates a proliferative response in human MCF7 breast cancer cells (2002) Biochem Biophys Res Commun, 293, pp. 622-628Dieudonne, M.N., Bussiere, M., Dos Santos, E., Adiponectin mediates antiproliferative and apoptotic responses in human MCF7 breast cancer cells (2006) Biochem Biophys Res Commun, 345, pp. 271-279Iyengar, P., Espina, V., Williams, T.W., Adipocyte-derived collagen VI affects early mammary tumor progression in vivo, demonstrating a critical interaction in the tumor/stroma microenvironment (2005) J Clin Invest, 115, pp. 1163-1176Lohsiriwat, V., Curigliano, G., Rietjens, M., Goldhirsch, A., Petit, J.Y., Autologous fat transplantation in patients with breast cancer: "silencing" or "fueling" cancer recurrence? (2011) Breast, 20, pp. 351-357Delay, E., Garson, S., Tousson, G., Sinna, R., Fat injection to the breast: Technique, results, and indications based on 880 procedures over 10 years (2009) Aesthet Surg J, 29, pp. 360-376Rietjens, M., De Lorenzi, F., Rossetto, F., Safety of fat grafting in secondary breast reconstruction after cancer (2011) J Plast Reconstr Aesthet Surg, 64, pp. 477-483Petit, J.Y., Lohsiriwat, V., Clough, K.B., The oncologic outcome and immediate surgical complications of lipofilling in breast cancer patients: A multicenter study-Milan-Paris-Lyon experience of 646 lipofilling procedures (2011) Plast Reconstr Surg, 128, pp. 341-346Rigotti, G., Marchi, A., Sbarbati, A., Adipose-derived mesenchymal stem cells: Past, present, and future (2009) Aesthetic Plast Surg, 33, pp. 271-273Elfadl, D., Garimella, V., Mahapatra, T.K., Lipomodelling of the breast: A review (2010) Breast, 19, pp. 202-209Mizuno, H., Hyakusoku, H., Fat grafting to the breast and adipose-derived stem cells: Recent scientific consensus and controversy (2010) Aesthet Surg J, 30, pp. 381-387Claro, Jr.F., Figueiredo, J.C., Zampar, A.G., Pinto-Neto, A.M., Applicability and safety of autologous fat for reconstruction of the breast (2012) Br J Surg, 99, pp. 768-780. , doi: 10.1002/bjs.8722Illouz, Y.G., Sterodimas, A., Autologous fat transplantation to the breast: A personal technique with 25 years of experience (2009) Aesthetic Plast Surg, 33, pp. 706-715Chan, C.W., McCulley, S.J., MacMillan, R.D., Autologous fat transfer-a review of the literature with a focus on breast cancer surgery (2008) J Plast Reconstr Aesthet Surg, 61, pp. 1438-1448Rosing, J.H., Wong, G., Wong, M.S., Sahar, D., Stevenson, T.R., Pu, L.L., Autologous fat grafting for primary breast augmentation: A systematic review (2011) Aesthetic Plast Surg, 35, pp. 882-890Pierrefeu-Lagrange, A.C., Delay, E., Guerin, N., Radiological evaluation of breasts reconstructed with lipomodeling (2006) Ann Chir Plast Esthet, 51, pp. 18-28Pulagam, S.R., Poulton, T., Mamounas, E.P., Long-term clinical and radiologic results with autologous fat transplantation for breast augmentation: Case reports and review of the literature (2006) Breast J, 12, pp. 63-65Veber, M., Tourasse, C., Toussoun, G., Moutran, M., Mojallal, A., Delay, E., Radiographic findings after breast augmentation by autologous fat transfer (2011) Plast Reconstr Surg, 127, pp. 1289-1299Carvajal, J., Patiño, J.H., Mammographic findings after breast augmentation with autologous fat injection (2008) Aesthet Surg J, 28, pp. 153-162Rubin, J.P., Coon, D., Zuley, M., Mammographic changes after fat transfer to the breast compared with changes after breast reduction: A blinded study (2012) Plast Reconstr Surg, 129, pp. 1029-1038Chala, L.F., De Barros, N., De Camargo Moraes, P., Fat necrosis of the breast: Mammographic, sonographic, computed tomography, and magnetic resonance imaging findings (2004) Curr Probl Diagn Radiol, 33, pp. 106-126Fraser, J.K., Hedrick, M.H., Cohen, S.R., Oncologic risks of autologous fat grafting to the breast (2011) Aesthet Surg J, 31, pp. 68-75Yoshimura, K., Sato, K., Aoi, N., Cell-assisted lipotransfer for cosmetic breast augmentation: Supportive use of adipose-derived stem/stromal cells (2008) Aesthetic Plast Surg, 32, pp. 48-55Petit, J.Y., Clough, K., Sarfati, I., Lipofilling in breast cancer patients: From surgical technique to oncologic point of view (2010) Plast Reconstr Surg, 126, pp. 262-263Rigotti, G., Marchi, A., Stringhini, P., Determining the oncological risk of autologous lipoaspirate grafting for post-mastectomy breast reconstruction (2010) Aesthetic Plast Surg, 34, pp. 475-480Petit, J.Y., Botteri, E., Lohsiriwat, V., Locoregional recurrence risk after lipofilling in breast cancer patients (2012) Ann Oncol, 23, pp. 582-588Manabe, Y., Toda, S., Miyazaki, K., Sugihara, H., Mature adipocytes, but not preadipocytes, promote the growth of breast carcinoma cells in collagen gel matrix culture through cancer-stromal cell interactions (2003) J Patholol, 201, pp. 221-228Yu, J.M., Jun, E.S., Bae, Y.C., Jung, J.S., Mesenchymal stem cells derived from human adipose tissues favor tumor cell growth in vivo (2008) Stem Cells Dev, 17, pp. 463-473Muehlberg, F.L., Song, Y.H., Krohn, A., Tissue-resident stem cells promote breast cancer growth and metastasis (2009) Carcinogenesis, 30, pp. 589-597Pearl, R.A., Leedham, S.J., Pacifico, M.D., The safety of autologous fat transfer in breast cancer: Lessons from stem cell biology (2012) J Plast Reconstr Aesthet Surg, 65, pp. 283-288Olumi, A.F., Grossfeld, G.D., Hayward, S.W., Carroll, P.R., Tlsty, T.D., Cunha, G.R., Carcinoma-associated fibroblasts direct tumor progression of initiated human prostatic epithelium (1999) Cancer Res, 59, pp. 5002-5011Perrot, P., Rousseau, J., Bouffaut, A.L., Safety concern between autologous fat graft, mesenchymal stem cell and osteosarcoma recurrence (2010) PLoS ONE, 5, pp. e10999Sun, B., Roh, K.H., Park, J.R., Therapeutic potential of mesenchymal stromal cells in a mouse breast cancer metastasis model (2009) Cytotherapy, 11, pp. 289-298. , 1 p following 29

Lignolytic-consortium omics analyses reveal novel genomes and pathways involved in lignin modification and valorization

BACKGROUND: Lignin is a heterogeneous polymer representing a renewable source of aromatic and phenolic bio-derived products for the chemical industry. However, the inherent structural complexity and recalcitrance of lignin makes its conversion into valuable chemicals a challenge. Natural microbial communities produce biocatalysts derived from a large number of microorganisms, including those considered unculturable, which operate synergistically to perform a variety of bioconversion processes. Thus, metagenomic approaches are a powerful tool to reveal novel optimized metabolic pathways for lignin conversion and valorization. RESULTS: The lignin-degrading consortium (LigMet) was obtained from a sugarcane plantation soil sample. The LigMet taxonomical analyses (based on 16S rRNA) indicated prevalence of Proteobacteria, Actinobacteria and Firmicutes members, including the Alcaligenaceae and Micrococcaceae families, which were enriched in the LigMet compared to sugarcane soil. Analysis of global DNA sequencing revealed around 240,000 gene models, and 65 draft bacterial genomes were predicted. Along with depicting several peroxidases, dye-decolorizing peroxidases, laccases, carbohydrate esterases, and lignocellulosic auxiliary (redox) activities, the major pathways related to aromatic degradation were identified, including benzoate (or methylbenzoate) degradation to catechol (or methylcatechol), catechol ortho-cleavage, catechol meta-cleavage, and phthalate degradation. A novel Paenarthrobacter strain harboring eight gene clusters related to aromatic degradation was isolated from LigMet and was able to grow on lignin as major carbon source. Furthermore, a recombinant pathway for vanillin production was designed based on novel gene sequences coding for a feruloyl-CoA synthetase and an enoyl-CoA hydratase/aldolase retrieved from the metagenomic data set. CONCLUSION: The enrichment protocol described in the present study was successful for a microbial consortium establishment towards the lignin and aromatic metabolism, providing pathways and enzyme sets for synthetic biology engineering approaches. This work represents a pioneering study on lignin conversion and valorization strategies based on metagenomics, revealing several novel lignin conversion enzymes, aromatic-degrading bacterial genomes, and a novel bacterial strain of potential biotechnological interest. The validation of a biosynthetic route for vanillin synthesis confirmed the applicability of the targeted metagenome discovery approach for lignin valorization strategies