Application of Thermomyces lanuginosus polygalacturonase produced in Komagataella phaffii in biomass hydrolysis and textile bioscouring.

In this work, the polygalacturonase (TL-PG1) from the thermophilic fungus Thermomyces lanuginosus was heterologously produced for the first time in the yeast Komagataella phaffii. The TL-PG1 was successfully expressed under the control of the AOX1 promoter and sequentially purified by His-tag affinity. The purified recombinant pectinase exhibited an activity of 462.6 U/mL toward polygalacturonic acid under optimal conditions (pH 6 and 55 ˚C) with a 2.83 mg/mL and 0.063 µmol/minute for Km and Vmax, respectively. When used as supplementation for biomass hydrolysis, TL-PG1 demonstrated synergy with the enzymatic cocktail Ctec3 to depolymerize orange citrus pulp, releasing 1.43 mg/mL of reducing sugar. In addition, TL-PG1 exhibited efficiency in fabric bioscouring, showing potential usage in the textile industry. Applying a protein dosage of 7 mg/mL, the time for the fabric to absorb water was 19.77 seconds (ten times faster than the control). Adding the surfactant Triton to the treatment allowed the reduction of the enzyme dosage by 50% and the water absorption time to 6.38 seconds. Altogether, this work describes a new versatile polygalacturonase from T. lanuginosus with the potential to be employed in the hydrolysis of lignocellulosic biomass and bioscouring.

Aqueous Extracts of Fermented Macrofungi Cultivated in Oilseed Cakes as a Carbon Source for Probiotic Bacteria and Potential Antibacterial Activity.

Plant biomass colonized by macrofungi can contain molecules with bioactive properties with applications to human/animal health. This work aimed to verify antibacterial activities from aqueous extracts from oil seed cakes of Jatropha curcas (JSC) and cottonseed (CSC), fermented by macrofungi for probiotic bacteria cultivation. Coriolopsis sp., Tyromyces sp., Panus lecomtei, and Pleurotus pulmonarius were cultivated in solid and submerged media. The aqueous extract of unfermented JSC was more efficient than glucose for the growth of all probiotic bacteria. Extracts from four macrofungi fermented in CSC favored Lactobacillus acidophilus growth. In solid fermentation, macrofungi extracts cultivated in JSC favored Bifidobacterium lactis growth. All fungi extracts showed more significant growth than carbohydrates among the four probiotic bacteria evaluated. Regarding antimicrobial activities, no fungal extract or bacterial supernatant showed a more significant inhibition halo for enteropathogenic bacteria than ampicillin (control). Extracts from P. lecomtei and Coriolopsis sp. in CSC showed inhibition halos for Salmonella enterica. Supernatants from L. acidophilus, B. lactis, and Lactobacillus rhamnosus resulted in more significant inhibition of Staphylococcus aureus than the control, which indicates possible antimicrobial activity. Unfermented JSC supernatant showed better results for bacterial growth, while supernatants and aqueous extracts from CSC fermentation can be used for probiotic bacteria culture.

The elusive Luminal B breast cancer and the mysterious chemokines.

PURPOSE: Invasive ductal breast cancer (IDC) is heterogeneous. Staging and immunohistochemistry (IH) allow for effective therapy but are not yet ideal. Women with Luminal B tumors show an erratic response to treatment. This prospective study with 81 women with breast cancer aims to improve the prognostic stratification of Luminal B patients. METHODS: This is a prospective translational study with 81 women with infiltrating ductal carcinoma, grouped by TNM staging and immunohistochemistry, for survival analysis, and their correlations with the chemokines. Serum measurements of 13 chemokines were performed, including 7 CC chemokines [CCL2(MCP1), CCL3(MIP1α), CCL4(MIP1ß), CCL5(Rantes), CCL11(Eotaxin), CCL17(TARC), CCL20(MIP3α)], 6 CXC chemokines [CXCL1(GroAlpha), CXCL5(ENA78), CCXCL8(IL-8), CXCL9(MIG), CXCL10(IP10), CXCL11(ITAC)]. RESULTS: Overall survival was significantly dependent on tumor staging and subtypes by immunohistochemistry, with a median follow-up time the 32.87 months (3.67-65.63 months). There were age correlations with IP10/CXCL10 chemokines (r = 0.4360; p = 0.0079) and TARC/CCL17 (Spearman + 0.2648; p = 0.0360). An inverse correlation was found between body weight and the chemokines Rantes/CCL5 (r = - 0.3098; p = 0.0169) and Eotaxin/CCL11 (r = - 0.2575; p = 0.0470). Smokers had a higher concentration of MIP3α/CCL20 (Spearman + 0.3344; p = 0.0267). Luminal B subtype patients who expressed lower concentrations of ENA78/CXCL5 (≤ 254.83 pg/ml) (Log-Rank p = 0.016) and higher expression of MIP1ß/CCL4 (> 34.84 pg/ml) (Log-Rank p = 0.014) had a higher risk of metastases. CONCLUSION: Patients with Luminal B breast tumors can be better stratified by serum chemokine expression, suggesting that prognosis is dependent on biomarkers other than TNM and IH.

TERT and BRAF V600E mutations in thyroid cancer of World Trade Center Responders.

The 2-fold excess thyroid cancer risk reported in multiple World Trade Center (WTC) disaster exposed cohorts cannot entirely be explained by surveillance and physician bias thus highlighting the need to investigate the potential consequences of the dust exposure, containing carcinogenic and endocrine disruptive elements, on the thyroid. This study investigated the presence of TERT promoter and BRAF V600E mutations in 20 WTC-exposed versus 23 matched non-exposed thyroid cancers as potential mechanism explaining the excess risk. Although no significant difference in BRAF V600E mutation was found, TERT promoter mutations were significantly more prevalent in WTC thyroid cancer versus non-exposed thyroid cancers (P = 0.021). The odds of a TERT promoter mutation was significantly higher in the WTC versus the non-WTC thyroid cancers after adjustment [ORadj: 7.11 (95% CI: 1.21-41.83)]. These results may indicate that exposure to the mixture of pollutants present in the WTC dust resulted in an excess thyroid cancer risk and potentially more aggressive thyroid cancer, warranting investigating WTC responders on thyroid-associated symptoms during their health checkups. Future studies should include long-term follow-up to provide important insights in whether thyroid-specific survival is negatively affected by WTC dust exposure and whether this is because of the presence of one or more driver mutations.

Multimodal evaluation of hemorrhages within intraretinal cystoid spaces in a blood-fluid level presentation.

INTRODUCTION: We describe characteristic findings on multimodal evaluation and the features of hemorrhage within a foveal cystoid space in a patient presenting cystoid macular edema secondary to Branch Retinal Vein Occlusion (BRVO). CASE DESCRIPTION: We report a case of a 64-year-old diabetic male patient presenting gradual blurry vision in the left eye. Fundoscopic findings were suggestive of BRVO, such as hard exudates and mild venous engorgement superotemporally and diffuse macular intraretinal hemorrhages. In the foveal area, there was cystoid edema with blood-fluid level (BFL) inside one of the cystoid spaces. Retina multimodal evaluation, including color, blue filter, and red-free fundus photography, fluorescein angiography, fundus autofluorescence, and spectral-domain optical coherence tomography (SD-OCT) B and C scan imaging, confirmed blood within foveal cystoid space. The patient underwent antiangiogenic therapy with significant improvement of macular edema and reduction of the cystoid space after 3 months. In addition, there was a resolution of visual symptoms. The cystoid space previously partially filled with blood, persisted, despite presenting smaller volume and medium reflectivity in the SD-OCT. CONCLUSIONS: Multimodal evaluation of blood-fluid level within foveal cystoid space in patients with BRVO has not been described previously. Identification of this sign may support the diagnosis of retinal vein occlusion in doubtful cases and further studies must be carried out to establish if the presence of BFL correlates with visual outcomes.

Advanced cell therapy with low tissue factor loaded product NestaCell® does not confer thrombogenic risk for critically ill COVID-19 heparin-treated patients.

Since the COVID-19 pandemic started, mesenchymal stromal cells (MSC) appeared as a therapeutic option to reduce the over-activated inflammatory response and promote recovery of lung damage. Most clinical studies use intravenous injection for MSC delivery, raising several concerns of thrombogenic risk due to MSC procoagulant activity (PCA) linked to the expression of tissue factor (TF/CD142). This is the first study that demonstrated procoagulant activity of TF+ human immature dental pulp stromal cells (hIDPSC, NestaCell® product) with the percentage of TF+ cells varied from 0.2% to 63.9% in plasma of healthy donors and COVID-19 heparin-treated patients. Thrombogenic risk of TF+ hIDPSCs was evaluated by rotational thromboelastometry (in vitro) and in critically ill COVID-19 patients (clinical trial). We showed that the thromboelastography is not enough to predict the risk of TF+ MSC therapies. Using TF-negative HUVEC cells, we demonstrated that TF is not a unique factor responsible for the cell's procoagulant activity. However, heparin treatment minimizes MSC procoagulant (in vitro). We also showed that the intravenous infusion of hIDPSCs with prophylactic enoxaparin administration in moderate to critically ill COVID-19 patients did not change the values of D-dimer, neither in the PT and PTT times. Our COVID-19 clinical study measured and selected the therapeutic cells with low TF (less than 25% of TF+ hIDPSCs). Our data indicate that the concomitant administration of enoxaparin and low TF-loaded is safe even for critically ill COVID-19 patients.

Bioactives and Extracellular Enzymes Obtained from Fermented Macrofungi Cultivated in Cotton and Jatropha Seed Cakes.

This work focused on obtaining fermented oil cake (cotton or Jatropha) via macrofungi growth with potential characteristics for animal feed formulations, such as the presence of extracellular enzymes, bioactive (ergosterol and antioxidants), and detoxification of antinutritional compounds. The concentration of phorbol esters was reduced by four macrofungi in Jatropha seed cake (JSC) to non-toxic levels. At least two macrofungi efficiently degraded free gossypol in cottonseed cake (CSC). Fermentation with Coriolopsis sp. INPA1646 and Tyromyces sp. INPA1696 resulted in increased ergosterol concentrations, antioxidant activity reduction, and high activity of laccases and proteases. Bromatological analysis indicated high crude protein concentrations, with partial solubilization by fungal proteases. Fermented products from Coriolopsis sp. and Tyromyces sp. in JSC or CSC can be considered important biological inputs for monogastric and polygastric animal feed.

Polypoidal Choroidal Vasculopathy Based on Non-ICGA Criteria in White Patients With Neovascular Age-Related Macular Degeneration.

PURPOSE: To determine prevalence of probable polypoidal choroidal vasculopathy (PCV) among White patients with neovascular age-related macular degeneration (nAMD) using non-indocyanine green angiography (ICGA) criteria DESIGN: Multicenter, multinational, retrospective, cross-sectional study. METHODS: A total of 208 treatment-naive eyes from Hispanic and non-Hispanic White individuals diagnosed with nAMD were included. All underwent color fundus photography (CFP), optical coherence tomography (OCT), and fluorescein angiography (FFA). De-identified images of study eyes were sent to 2 groups of graders. Group 1 reviewed CFP, OCT, and FFA to confirm nAMD diagnosis. Group 2 reviewed CFP and OCT to determine highly suggestive features for PCV. Probable PCV diagnosis defined as the presence of ≥2 of 4 highly suggestive features for PCV: notched or fibrovascular pigment epithelial detachment (PED) on CFP, sharply-peaked PED, notched PED, and hyperreflective ring on OCT. RESULTS: Eleven eyes were excluded because of poor image quality (6) or non-nAMD diagnosis (5). Of 197 eligible eyes (197 patients), the mean age (SD) was 78.8 years (8.9), 44.2% were men, 26.4% were Hispanic, and 73.6% were non-Hispanic White individuals; 41.1%, 23.4%, 9.1%, and 2.5% had ≥1, ≥2, ≥3, and 4 highly suggestive features. Results showed that 23.4% (95% CI, 17.6%-29.9%) had probable PCV diagnosis. Predominantly occult CNV was more frequently found in probable PCV than nAMD subgroup (84.8% vs 64.9%, P = .01). Hispanic White individuals had a lower prevalence of probable PCV than non-Hispanic White individuals (9.6% vs 28.2%, P = .006) CONCLUSIONS: These findings suggest that probable PCV occurs between 17.6% and 29.9% in White individuals with nAMD, and more commonly in non-Hispanic than in Hispanic White individuals.

General Practice/Family Medicine Portuguese doctors: “technicists” or “systemics”? / Medicina General/Medicina de Familia. Médicos portugueses: ¿«técnicos» o «sistémicos»?

To study if the consultation's problems classification of a fictitious case by General and Family Medicine doctors, showed the characteristics of being a mechanistic or a systemic approach. Exploratory cross-sectional observational study in a convenience sample of the General Practice/Family Medicine population, internees included, in April 2020, applying a modified real world clinical case. Central Portugal and the Autonomous Region of Azores. General Practice/Family Medicine specialists and internees. Electronic Doctors invitation to participate, anonymously, in specific doctor's social networks. A self-fulfilling questionnaire was used to verify the classification of a clinical case in Subjective, Assessment and Plan (from the SOAP methodology) with the response options of the International Classification of Primary Health Care (ICPC2) in chapters P (Psychological) and Z (Social) possible for this case. “Technicists” doctors, only classifying “P” codes and “Systemics” classifying “P+Z” or only “Z” ICPC2 codes were defined. Differences between genders, work place, being an internee or specialist and being a tutor in specialized formation were studied. A sample of 227 30% (n=68) males, specialists represented 66% (n=149), of whom 49% (n=73) were internee's tutors and 34% (n=78) were internees, was studied. In the Subjective chapter of the SOAP methodology, 44.1% (n=100) were “technicists”, for Assessment n=93 (40.8%) were “technicists” and for P chapter classification 56.8% were “technicists”. For S, A and P chapters classification there was no significant difference between the considered variables. In this sample General Practice/Family Medicine Portuguese doctors were more “systemic” for the S and A chapters of the SOAP model, And “technicists” in the P chapter.(AU)

El enfoque de la medicina centrada en el paciente (MCP) es cada vez más importante debido a su asociación de efectos positivos sobre el médico y el paciente. El presente estudio tuvo como objetivo conocer si el abordaje de un caso clínico, a través de la clasificación de problemas de consulta de un caso ficticio por médicos de Medicina General y de Familia, presenta características de abordaje mecanicista o sistémico. Estudio observacional transversal en una muestra de conveniencia de la población de Medicina General/Medicina Familiar, incluidos los internos, en abril del 2020, aplicando un caso clínico del mundo real modificado de Portugal y los especialistas en Medicina General/Medicina Familiar de Azores y de Portugal central. Especialistas en Medicina General/Medicina Familiar e internados. Los médicos fueron invitados a participar, de forma anónima, mediante invitación electrónica, ya sea en la red oficial de pasantías o en la red social de un médico específico, la red MGFamiliar. Se utilizó un cuestionario autocumplimentable para verificar la clasificación de un caso clínico en subjetivo, evaluación y planificación (de la metodología SOAP) con las opciones de respuesta de la Clasificación Internacional de Atención Primaria de Salud (CIPC2) en los capítulos P (Psicológico) y Z (Social) posible para este caso. Se definieron médicos «técnicos» solo clasificando con códigos «P» y «sistémicos» clasificando «P+Z»’ o solo con códigos ICPC2 «Z». Se estudiaron las diferencias entre géneros, lugar de trabajo, ser interno o especialista, ser tutor en formación especializada. En una muestra de 227 sujetos, 30% (n=68) varones, los especialistas representaron el 66% (n=149), de los cuales el 49% (n=73) fueron tutores internos y el 34% (n=78), internos. Para la clasificación en el capítulo subjetivo de la metodología SOAP, el 44,1% (n=100) eran «técnicos», para la evaluación n=93 (40,8%) eran «técnicos» y para la clasificación del capítulo P el 56,8% eran «técnicos».(AU)

General Practice/Family Medicine Portuguese doctors: "technicists" or "systemics"?

To study if the consultation's problems classification of a fictitious case by General and Family Medicine doctors, showed the characteristics of being a mechanistic or a systemic approach. Exploratory cross-sectional observational study in a convenience sample of the General Practice/Family Medicine population, internees included, in April 2020, applying a modified real world clinical case. Central Portugal and the Autonomous Region of Azores. General Practice/Family Medicine specialists and internees. Electronic Doctors invitation to participate, anonymously, in specific doctor's social networks. A self-fulfilling questionnaire was used to verify the classification of a clinical case in Subjective, Assessment and Plan (from the SOAP methodology) with the response options of the International Classification of Primary Health Care (ICPC2) in chapters P (Psychological) and Z (Social) possible for this case. "Technicists" doctors, only classifying "P" codes and "Systemics" classifying "P+Z" or only "Z" ICPC2 codes were defined. Differences between genders, work place, being an internee or specialist and being a tutor in specialized formation were studied. A sample of 227 30% (n=68) males, specialists represented 66% (n=149), of whom 49% (n=73) were internee's tutors and 34% (n=78) were internees, was studied. In the Subjective chapter of the SOAP methodology, 44.1% (n=100) were "technicists", for Assessment n=93 (40.8%) were "technicists" and for P chapter classification 56.8% were "technicists". For S, A and P chapters classification there was no significant difference between the considered variables. In this sample General Practice/Family Medicine Portuguese doctors were more "systemic" for the S and A chapters of the SOAP model, And "technicists" in the P chapter.

Análise da polifarmácia e de fatores associados ao uso racional de medicamentos em idosos moradores de residência de cuidados de longa permanência / Analysis of polypharmacy and factors associated with the rational use of medicines in elderly residents of long-term care residences

Este trabalho teve por objetivo analisar a polifarmácia e os fatores associados ao uso racional de medicamentos em idosos moradores de residência de cuidado de longa permanência na perspectiva do cuidado nas condições crônicas de saúde. Foi desenvolvido junto a 68 prontuários, prescrições médicas e/ou registros de administração de medicamentos de idosos moradores de um residencial filantrópico do Vale do Paranapanema/São Paulo. A coleta de dados foi realizada por meio de um instrumento com perguntas abertas e fechadas sobre a temática e a análise se deu por meio do cálculo de frequência absoluta e relativa. A polifarmácia fazia parte da vida de grande parte dos idosos. Não foi possível compreender, por falta de sistematização e de controle documental interno, como os responsáveis técnicos do setor administrativo e da saúde adquiriam medicamentos, nem os custos envolvidos com a compra e o desperdício. Os medicamentos e seus resíduos eram descartados no lixo orgânico. Não foi possível também determinar se idosos recebiam o medicamento apropriado às suas necessidades clínicas, na dose e no período adequado às suas condições de saúde. Conclui-se que idosos moradores do residencial estão expostos à polifarmácia e o uso racional de medicamentos não é praticado.

This study aimed to analyze polypharmacy and factors associated with the rational use of medicines in elderly residents of long-term care homes from the perspective of care in chronic health conditions. This study was developed with sixty-eight medical records, medical prescriptions, and/or medication administration records of elderly residents of a philanthropic home, located in a municipality in the Vale do Paranapanema/São Paulo. Data was collected using an instrument with open and closed questions on the theme. The analysis consisted of calculating absolute and relative frequency. Polypharmacy was part of the life of most elderly people. It was not possible to understand, due to the lack of systematization and internal document control, how the technicians responsible for the administrative and health sectors acquired the medicines, nor the costs involved with the purchase and waste. Medicines and their residues were disposed of in organic waste. It was also not possible to determine whether the elderly received medication appropriate to their clinical needs, in the dose and within the appropriate period to their health conditions. Therefore, elderly residents of the residential are exposed to polypharmacy and the rational use of medicines is not practiced.

Advanced cell therapy with low tissue factor loaded product NestaCell® does not confer thrombogenic risk for critically ill COVID-19 heparin-treated patients

Since the COVID-19 pandemic started, mesenchymal stromal cells (MSC) appeared as a therapeutic option to reduce the over-activated inflammatory response and promote recovery of lung damage. Most clinical studies use intravenous injection for MSC delivery, raising several concerns of thrombogenic risk due to MSC procoagulant activity (PCA) linked to the expression of tissue factor (TF/CD142). This is the first study that demonstrated procoagulant activity of TF+ human immature dental pulp stromal cells (hIDPSC, NestaCell® product) with the percentage of TF+ cells varied from 0.2% to 63.9% in plasma of healthy donors and COVID-19 heparin-treated patients. Thrombogenic risk of TF+ hIDPSCs was evaluated by rotational thromboelastometry (in vitro) and in critically ill COVID-19 patients (clinical trial). We showed that the thromboelastography is not enough to predict the risk of TF+ MSC therapies. Using TF-negative HUVEC cells, we demonstrated that TF is not a unique factor responsible for the cell's procoagulant activity. However, heparin treatment minimizes MSC procoagulant (in vitro). We also showed that the intravenous infusion of hIDPSCs with prophylactic enoxaparin administration in moderate to critically ill COVID-19 patients did not change the values of D-dimer, neither in the PT and PTT times. Our COVID-19 clinical study measured and selected the therapeutic cells with low TF (less than 25% of TF+ hIDPSCs). Our data indicate that the concomitant administration of enoxaparin and low TF-loaded is safe even for critically ill COVID-19 patients.

Exosomes in the Tumor Microenvironment: From Biology to Clinical Applications.

Cancer is one of the most important health problems and the second leading cause of death worldwide. Despite the advances in oncology, cancer heterogeneity remains challenging to therapeutics. This is because the exosome-mediated crosstalk between cancer and non-cancer cells within the tumor microenvironment (TME) contributes to the acquisition of all hallmarks of cancer and leads to the formation of cancer stem cells (CSCs), which exhibit resistance to a range of anticancer drugs. Thus, this review aims to summarize the role of TME-derived exosomes in cancer biology and explore the clinical potential of mesenchymal stem-cell-derived exosomes as a cancer treatment, discussing future prospects of cell-free therapy for cancer treatment and challenges to be overcome.

Mebendazole disrupts stromal desmoplasia and tumorigenesis in two models of pancreatic cancer.

The five-year survival rate for metastatic pancreatic cancer is currently only 3%, which increases to 13% with local invasion only and to 39% with localized disease at diagnosis. Here we evaluated repurposed mebendazole, an approved anthelminthic drug, to determine how mebendazole might work at the different stages of pancreatic cancer formation and progression. We asked if mebendazole could prevent initiation of pancreatic intraepithelial neoplasia precursor lesions, interfere with stromal desmoplasia, or suppress tumor growth and liver metastasis. In both the Kras LSL.G12D/+; Pdx1-Cre (KC) mouse model of caerulein-induced inflammatory pancreatitis and the Kras LSL.G12D/+; Tp53 R172H/+; Pdx1-Cre (KPC) mouse model of advanced pancreatic cancer, mebendazole significantly reduced pancreas weight, dysplasia and intraepithelial neoplasia formation, compared to controls. Mebendazole significantly reduced trichrome-positive fibrotic connective tissue and α-SMA-positive activated pancreatic stellate cells that heralds fibrogenesis. In the aggressive KPC model, mebendazole significantly suppressed pancreatic tumor growth, both as an early and late intervention. Mebendazole reduced the overall incidence of pancreatic cancer and severity of liver metastasis in KPC mice. Using early models of pancreatic cancer, treatment with mebendazole resulted in less inflammation, decreased dysplasia, with the later stage model additionally showing a decreased tumor burden, less advanced tumors, and a reduction of metastasis. We conclude that mebendazole should be investigated further as a component of adjuvant therapy to slow progression and prevent metastasis, and well as for primary prevention in the highest risk patients.

BDNF as a Putative Target for Standardized Extract of Ginkgo biloba-Induced Persistence of Object Recognition Memory.

Despite considerable progress on the study of the effect of standardized extract of Gingko biloba (EGb) on memory processes, our understanding of its role in the persistence of long-term memory (LTM) and the molecular mechanism underlying its effect, particularly episodic-like memory, is limited. We here investigated the effects of EGb on the long-term retention of recognition memory and its persistence and BDNF expression levels in the dorsal hippocampal formation (DHF). Adult male Wistar rats (n = 10/group) were handled for 10 min/5 day. On day 6, the animals were treated with vehicle or 0.4 mg/kg diazepam (control groups) or with EGb (250, 500 or 100 mg/kg) 30 min before the training session (TR1), in which the animals were exposed to two sample objects. On day 7, all rats underwent a second training session (TR2) as described in the TR1 but without drug treatment. Object recognition memory (ORM) was evaluated on day 8 (retention test, T1) and day 9 (persistence test, T2). At the end of T1or T2, animals were decapitated, and DHF samples were frozen at -80 °C for analyses of the differential expression of BDNF by Western blotting. EGb-treated groups spent more time exploring the novel object in T2 and showed the highest recognition index (RI) values during the T1 and T2, which was associated with upregulation of BDNF expression in the DHF in a dose-and session-dependent manner. Our data reveal, for the first time, that EGb treatment before acquisition of ORM promotes persistence of LTM by BDNF differential expression.

Operable breast cancer: How not to worsen the prognosis, especially in triple negative and stage II tumors.

INTRODUCTION: Oncological surgery must follow some fundamental principles to be truly curative, one of which is the resection of the tumor with surgical margins free of neoplasia. In breast cancer, surgery with positive margins should be expanded immediately. There are probably different intensities, between the stages and molecular subtypes of operable breast cancer, of worsening prognosis due to the surgical margin compromised by the neoplasia in women not submitted to the necessary enlargement of the positive surgical margin. MATERIALS AND. METHODS: Seven hundred and forty-seven women with invasive ductal carcinoma of the breast, analyzing anatomical-pathological information, types of surgery, molecular subtypes, and the presence or absence of the surgical margin compromised by neoplasia. RESULTS: Sixty-one (8.2%) patients had positive surgical margin, causing 2.85 times more risk of locoregional relapse compared to negative surgical margin by multivariate analysis. In subgroup analysis, among stages I, II and III, stage II was the most negatively impacted, with those patients presenting 2.42 times more risk of distant metastasis and 4.94 times more risk of locoregional relapses compared to negative surgical margin by multivariate analysis. Among the molecular subtypes, Triple Negative tumors with a positive surgical margin had 3.56 times more risk of death, 4.98 times more risk of distant metastasis and 5.55 times more risk of locoregional relapse compared to negative surgical margin by multivariate analysis. CONCLUSIONS: The positive surgical margin, especially in Stage II and Triple-Negative breast cancer patients negatively impact the patient's evolution, increasing risk of distant metastasis and death.

Comprehensive Assessment of Copy Number Alterations Uncovers Recurrent AIFM3 and DLK1 Copy Gain in Medullary Thyroid Carcinoma.

Medullary thyroid carcinoma (MTC) is a malignant tumor originating from thyroid C-cells that can occur either in sporadic (70-80%) or hereditary (20-30%) form. In this study we aimed to identify recurrent copy number alterations (CNA) that might be related to the pathogenesis or progression of MTC. We used Affymetrix SNP array 6.0 on MTC and paired-blood samples to identify CNA using PennCNV and Genotyping Console software. The algorithms identified recurrent copy number gains in chromosomes 15q, 10q, 14q and 22q in MTC, whereas 4q cumulated losses. Coding genes were identified within CNA regions. The quantitative PCR analysis performed in an independent series of MTCs (n = 51) confirmed focal recurrent copy number gains encompassing the DLK1 (14q32.2) and AIFM3 (22q11.21) genes. Immunohistochemistry confirmed AIFM3 and DLK1 expression in MTC cases, while no expression was found in normal thyroid tissues and few MTC samples were found with normal copy numbers. The functional relevance of CNA was also assessed by in silico analysis. CNA status correlated with protein expression (DLK1, p = 0.01), tumor size (DLK1, p = 0.04) and AJCC staging (AIFM3p = 0.01 and DLK1p = 0.05). These data provide a novel insight into MTC biology, and suggest a common CNA landscape, regardless of if it is sporadic or hereditary MTC.

Exosomes in the tumor microenvironment: from biology to clinical applications

Cancer is one of the most important health problems and the second leading cause of death worldwide. Despite the advances in oncology, cancer heterogeneity remains challenging to therapeutics. This is because the exosome-mediated crosstalk between cancer and non-cancer cells within the tumor microenvironment (TME) contributes to the acquisition of all hallmarks of cancer and leads to the formation of cancer stem cells (CSCs), which exhibit resistance to a range of anticancer drugs. Thus, this review aims to summarize the role of TME-derived exosomes in cancer biology and explore the clinical potential of mesenchymal stem-cell-derived exosomes as a cancer treatment, discussing future prospects of cell-free therapy for cancer treatment and challenges to be overcome.

Paper Spray Ionization Mass Spectrometry as a Potential Tool for Early Diagnosis of Cervical Cancer.

Squamous intraepithelial lesion is an abnormal growth of epithelial cells on the surface of the cervix that may lead to cervical cancer. Analytical protocols for the determination of squamous intraepithelial lesions are in high demand, since cervical cancer is the fourth most diagnosed cancer among women in the world. Here, paper spray ionization mass spectrometry (PSI-MS) is used to distinguish between healthy (negative for intraepithelial lesion or malignancy) and diseased (high-grade squamous intraepithelial lesion) blood plasmas. A total of 86 blood samples of different women (49 healthy samples, 37 diseased samples) were collected, and the plasmas were prepared. Then, 10 µL of each plasma sample was deposited onto triangular papers for PSI-MS analysis. No additional step of sample preparation was necessary. The interval-successive projection algorithm linear discriminant analysis (iSPA-LDA) was applied to the PSI mass spectra, showing six ions (mostly phospholipids) that were predictive of healthy and diseased plasmas. Values of 77% accuracy, 86% sensitivity, 80% positive predictive value (PPV), and 75% negative predictive value (NPV) were achieved. This study provides evidence that PSI-MS may potentially be used as a fast and simple analytical technique for the early diagnosis of cervical cancer.

Population pharmacokinetics and biodistribution of benznidazole in mice.

OBJECTIVES: To evaluate the population pharmacokinetics of different benznidazole treatment regimens and the drug's biodistribution in mice. METHODS: Two hundred mice were divided into five groups according to benznidazole dosing regimens: (1) 100 mg/kg/day for 20 days; (2) 100 mg/kg/day for 40 days; (3) 200 mg/kg/day for 20 days; (4) 40 mg/kg/day for 20 days; or (5) 40 mg/kg/day for 40 days. The mice were euthanized and blood, heart, liver, colon and brain were collected. Samples were prepared by liquid-liquid extraction and analysed by HPLC-diode-array detection. The pharmacokinetic analysis of benznidazole was evaluated via non-linear mixed-effects modelling using the NONMEN program. RESULTS: Our results demonstrate that mouse weight allometrically influences benznidazole clearance; the AUC curve and the highest plasma concentration are dose proportional; benznidazole does not influence its own metabolism; its tissue distribution is limited; and the standard treatment regimen for Chagas' disease in mice (100 mg/kg/day for 20 days) is inadequate from a pharmacokinetic standpoint, as are the other regimens tested in this study (100 mg/kg/day for 40 days, 200 mg/kg/day for 20 days and 40 mg/kg/day for 20 or 40 days). CONCLUSIONS: Benznidazole reformulations that allow better tissue penetration and plasma and tissue exposure should be evaluated to enable higher cure rates in both animals and patients. The population pharmacokinetic model developed here can allow optimization of the dosing regimen of benznidazole to treat experimental Chagas' disease. Determining appropriate treatment regimens in animals allows translation of these to clinical studies.