Functional purification of the monocarboxylate transporter of the yeast Candida utilis

Plasma membranes of the yeast, Candida utilis, were solubilized with octyl-b-D- lucopyranoside and a fraction enriched in the lactate carrier was obtained with DEAE-Sepharose anion- exchange chromatography, after elution with 0.4 M NaCl. The uptake of lactic acid into proteoliposomes, containing the purified protein fraction and cytochrome c oxidase, was dependent on a proton-motive force and the transport specificity was consistent with the one of C. utilis intact cells. Overall, we have obtained a plasma membrane fraction enriched in the lactate carrier of C. utilis in which the transport properties were preserved. Given the similarities between the lactate transport of C. utilis and the one of mammalian cells, this purified system could be further explored to screen for specific lactate inhibitors, with potential therapeutic applications

Propolis: A complex natural product with a plethora of biological activities that can be explored for drug development

The health industry has always used natural products as a rich, promising, and alternative source of drugs that are used in the health system. Propolis, a natural resinous product known for centuries, is a complex product obtained by honey bees from substances collected from parts of different plants, buds, and exudates in different geographic areas. Propolis has been attracting scientific attention since it has many biological and pharmacological properties, which are related to its chemical composition. Several in vitro and in vivo studies have been performed to characterize and understand the diverse bioactivities of propolis and its isolated compounds, as well as to evaluate and validate its potential. Yet, there is a lack of information concerning clinical effectiveness. The goal of this review is to discuss the potential of propolis for the development of new drugs by presenting published data concerning the chemical composition and the biological properties of this natural compound from different geographic origins

Microbes and cancer: friends or faux?

Cancer is one of the most aggressive and deadly diseases in the world, representing the second leading cause of death. It is a multifactorial disease, in which genetic alterations play a key role, but several environmental factors also contribute to its development and progression. Infections induced by certain viruses, bacteria, fungi and parasites constitute risk factors for cancer, being chronic infection associated to the development of certain types of cancer. On the other hand, susceptibility to infectious diseases is higher in cancer patients. The state of the host immune system plays a crucial role in the susceptibility to both infection and cancer. Importantly, immunosuppressive cancer treatments increase the risk of infection, by decreasing the host defenses. Furthermore, alterations in the host microbiota is also a key factor in the susceptibility to develop cancer. More recently, the identification of a tumor microbiota, in which bacteria establish a symbiotic relationship with cancer cells, opened a new area of research. There is evidence demonstrating that the interaction between bacteria and cancer cells can modulate the anticancer drug response and toxicity. The present review focuses on the interaction between microbes and cancer, specifically aiming to: (1) review the main infectious agents associated with development of cancer and the role of microbiota in cancer susceptibility; (2) highlight the higher vulnerability of cancer patients to acquire infectious diseases; (3) document the relationship between cancer cells and tissue microbiota; (4) describe the role of intratumoral bacteria in the response and toxicity to cancer therapy.This research was funded by National Funds through FCT—Fundação para a Ciência e a Tecnologia, I.P., within CINTESIS, R&D Unit, grant number UIDB/4255/2020

Targeting oncogenic microRNAs in triple negative breast cancer using CRISPR/cas9 approach

info:eu-repo/semantics/publishedVersio

Microbiota-derived short-chain fatty acids: new road in colorectal cancer therapy

The colon microbiota is an important player in colorectal cancer (CRC) development, which is responsible for most of the cancer-related deaths worldwide. During carcinogenesis, the colon microbiota composition changes from a normobiosis profile to dysbiosis, interfering with the production of short-chain fatty acids (SCFAs). Each SCFA is known to play a role in several biological processes but, despite their reported individual effects, colon cells are exposed to these compounds simultaneously and the combined effect of SCFAs in colon cells is still unknown. Our aim was to explore the effects of SCFAs, alone or in combination, unveiling their biological impact on CRC cell phenotypes. We used a mathematical model for the prediction of the expected SCFA mixture effects and found that, when in mixture, SCFAs exhibit a concentration addition behavior. All SCFAs, alone or combined at the physiological proportions founded in the human colon, revealed to have a selective and anticancer effect by inhibiting colony formation and cell proliferation, increasing apoptosis, disturbing the energetic metabolism, inducing lysosomal membrane permeabilization, and decreasing cytosolic pH. We showed for the first time that SCFAs are specific towards colon cancer cells, showing promising therapeutic effects. These findings open a new road for the development of alternatives for CRC therapy based on the increase in SCFA levels through the modulation of the colon microbiota composition.This work was supported by the project EcoAgriFood (NORTE-01-0145-FEDER-00009), supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). Sara Gomes thanks the FCT for her PhD grant (SFRH/BD/140965/2018). This work was financed by the Portuguese Foundation for Science and Technology (Fundação para a Ciência e a Tecnologia, FCT) within the scope of project PTDC/QUI-IN/28662/2017. This work was also supported by the strategic programme UID/BIA/04050/2019, funded by national funds through the FCT I.P

How to decipher a microbial puzzle on microbial control: hands on

How to decipher a microbial puzzle on microbial control: hands onAntibiotics are among the most commonly prescribed drugs, but they are often misused, contributing to bacterial resistance, being an important public health problem, with clinical/economic impacts. Aiming to contribute to convey information to students on Microbiology/antibiotic use, we developed different teaching activities, with 6th and 9th grade students. To assess the impact of these activities and student learning, we applied a pre-validated questionnaire before and after completion of the activities (pre- and post-test). We observed that most students had preconceived ideas that antibiotics were effective against viruses, however, after the implemented activities, the performance of students in the post-test improved significantly for both student grades. Regarding the use of antibiotics in specific diseases, students showed lack of knowledge concerning the treatment of cold, flu and tuberculosis, with some improvement in the post-test. Concerning antibiotic use, the initial knowledge of the 9th grade students was better than the 6th grade, nevertheless the highest improvement in knowledge was observed for the 6th grade students. Our results support the use of teaching activities on Microbiology/antibiotic use that promote the development of critical thinking, analyze and solve problems. Our results support the need to educate students/communities on the proper use of antibiotics

Portuguese students' knowledge of antibiotics: a cross-sectional study of secondary school and university students in Braga

<p>Abstract</p> <p>Background</p> <p>Recent surveys show that the knowledge of the general public about the correct use of antibiotics is limited. This contributes to the problem of inappropriate antibiotic use, leading to a progressive loss of bacterial sensitivity to these drugs and the spreading of resistant strains of bacteria.</p> <p>Methods</p> <p>In this cross-sectional study, a questionnaire about antibiotic use was given to a sample of students in the 9^thand 12^thgrades of secondary school and in the first year of university in the north of Portugal.</p> <p>Results</p> <p>349 students returned completed questionnaires. Deficits were found in the students' knowledge of antibiotics and their correct use. Only 4% of 9^thgrade students were aware that antibiotics are used to treat bacteria only, while 14% of 12^thgrade students and 29% of first-year university students were aware of this. Fewer students were aware that antibiotics are used to treat tuberculosis. There were deficiencies in the knowledge of timing and duration of therapy. However close to 70% of these students are aware that inappropriate use of antibiotics can contribute to resistance to these drugs.</p> <p>Conclusion</p> <p>This study has observed a lack of general knowledge on correct antibiotic use in Portugal, as has been found in other countries. Since this may be due to a lack of formal education on this subject, we believe that a teaching unit on infectious diseases should be included in the 9^thand 12^thgrades, in all curricular areas, with emphasis on bacterial and viral pathogens and correct antibiotic use. In addition, education on the correct use of medications may need to begin at much earlier ages.</p

A hospital based cohort study of colorectal cancer cases treated at Braga Hospital, Northern Portugal

Background: Colorectal cancer (CRC) is the third most common cancer and the fourth most frequent cause of cancer death worldwide. Nonetheless, despite being a frequent cancer on which many epidemiological international studies have already been written, Portuguese epidemiological data are scarce and in particular there are very few specific data for Minho Region, which is traditionally recognized as a high incidence area. Aim: Characterize CRC patients treated at Braga Hospital. Methods: Data regarding clinical and preoperative diagnostic examinations, operative reports and histopathological and follow-up data was collected prospectively and stored in two Excel PC databases (colon and rectal cancer) and statistically analysed using the Statistical Package for the Social Sciences, version 19.0 (SPSS Inc., Chicago, Illinois, USA). All comparisons were examined for statistical significance using Pearson’s chi-square (?2) test and Fisher’s exact test (when n<5), with the threshold for significance P values <0.05. Overall survival and Survival free disease were both assessed using the Kaplan-Meier method. Results: The study comprises 672 patients with histological diagnosis of CRC treated in Braga Hospital between 2005 and 2009. It included 62.3% males and 37.7% females and most patients (60.5%) were between 61-80 years old. 65.3% of the cases arose from colon cancer and 34.7% from rectal cancer. We observed that 94.8% of the patients had no previous history of colorectal polyps. 4.1% had a previous personal history of CRC and 7.7% of a different cancer. 9.7% had a positive CRC family history. Most patients (81.3%) were symptomatic at diagnosis, while 18.8% were detected by routine colonoscopies. Colon and rectal cancer from most patients was at IIA stage and IV stage respectively. Follow-up time ranged between 1 and 5 years and, during this period, 26.7% of colon cancer patients and 25.3% of rectal cancer patients died from a colorectal cancer-related cause; also, 14.6% and 19.3% respectively had recurrence, mainly in the liver. Conclusion: This is the first study of a large cohort of CRC patients from the Minho Region in Northern Portugal. The large majority of the 672 cases were diagnosed because symptomatic and at an advanced stage, with a relatively poor prognosis. These findings emphasize the need to start a screening program and diagnose CRC at an early stage, thus increasing cure rates and improving resource management

Disruption of pH dynamics suppresses proliferation and potentiates doxorubicin cytotoxicity in breast cancer cells

The reverse pH gradient is a major feature associated with cancer cell reprogrammed metabolism. This phenotype is supported by increased activity of pH regulators like ATPases, carbonic anhydrases (CAs), monocarboxylate transporters (MCTs) and sodium–proton exchangers (NHEs) that induce an acidic tumor microenvironment, responsible for the cancer acid-resistant phenotype. In this work, we analyzed the expression of these pH regulators and explored their inhibition in breast cancer cells as a strategy to enhance the sensitivity to chemotherapy. Expression of the different pH regulators was evaluated by immunofluorescence and Western blot in two breast cancer cell lines (MDA-MB-231 and MCF-7) and by immunohistochemistry in human breast cancer tissues. Cell viability, migration and invasion were evaluated upon exposure to the pH regulator inhibitors (PRIs) concanamycin-A, cariporide, acetazolamide and cyano-4-hydroxycinnamate. Additionally, PRIs were combined with doxorubicin to analyze the effect of cell pH dynamic disruption on doxorubicin sensitivity. Both cancer cell lines expressed all pH regulators, except for MCT1 and CAXII, only expressed in MCF-7 cells. There was higher plasma membrane expression of the pH regulators in human breast cancer tissues than in normal breast epithelium. Additionally, pH regulator expression was significantly associated with different molecular subtypes of breast cancer. pH regulator inhibition decreased cancer cell aggressiveness, with a higher effect in MDA-MB-231. A synergistic inhibitory effect was observed when PRIs were combined with doxorubicin in the breast cancer cell line viability. Our results support proton dynamic disruption as a breast cancer antitumor strategy and the use of PRIs to boost the activity of conventional therapy.This research was funded by National funds, through the Foundation for Science and Technology (FCT)—project UIDB/50026/2020 and UIDP/50026/2020; and by the projects NORTE 01-0145-FEDER-000013 and NORTE-01-0145-FEDER-000023, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). This work was also supported by an internal CE SPU project MetabRes_CESPU_2017. DT-V received a fellowship from FCT (ref. SFRH/BD/103025/2014)