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Prostate cancer is a very prevalent disease in men, especially in Western countries. The treatment of this neoplasm, both localized and locally advanced, is based on the clinical risk analysis (Gleason, tumor size, PSA, and other factors) and is founded on surgery and/or radiotherapy with or without androgen blockade with a GnRH analog (hormone gonadotropin releaser). However, in patients who invariably progress to a metastatic disease scenario, the tumors may present a heterogeneous behavior, depending on whether or not they are sensitive to androgen blockade therapy. Due to the poor prognosis of the metastatic castration-resistant scenario, current research carried out in the molecular biology and genetics field has identified several gene alterations associated with the development of prostate cancer, which correlate with clinical risk, therapeutic predictive responses, and prognosis. Among the associated gene alterations, the genes of the DNA repair pathway are correlated with diseases that present: a higher risk of recurrence; early metastasis; worse cancer-specific survival; familial risk, and predictive responses to new targeted therapies. Therefore, the breast cancer susceptibility genes, BRCA1 and BRCA2 (and other variants), present in the DNA repair machinery are being investigated to provide more (and better) therapeutic options for the treatment of the disease in the advanced scenario. This review was aimed to describe the malignant prostate disease, especially with regard to DNA repair mechanisms, genomic analysis of prostate cancer, predictive and prognostic implications, as well as on the development of poly-(ADP-ribose) polymerase (PARP) inhibitors, synthetic lethality mechanisms, and BRCAness phenomenon.

It is well known that the new coronavirus pandemic has global environmental, public health, and economic implications. In this sense, this study aims to monitor SARS-CoV-2 in the largest wastewater treatment plant of Goiânia, which processes wastewater from more than 700,000 inhabitants, and to correlate the molecular and clinical data collected. Influent and effluent samples were collected at Dr. Helio de Seixo Britto's wastewater treatment plant from January to August 2021. Viral concentration was performed with polyethylene glycol before viral RNA extraction. Real-time qPCR (N1 and N2 gene assays) was performed to detect and quantify the viral RNA present in the samples. The results showed that 43.63% of the samples were positive. There is no significant difference between the detection of primers N1 (mean 3.23 log10 genome copies/L, std 0.23) and N2 (mean 2.95 log10 genome copies/L, std 0.29); also, there is no significant difference between the detection of influent and effluent samples. Our molecular data revealed a positive correlation with clinical data, and infection prevalence was higher than clinical data. In addition, we developed a user-friendly web application to predict the number of infected people based on the detection of viral load present in wastewater samples and may be applied as a public policy strategy for monitoring ongoing outbreaks.

Objective To evaluate the secondary attack rate (SAR) in children and adolescents, contacts of essential activities workers who were infected by SARS‐CoV‐2; and to describe associated clinical and epidemiological data. Methods A cross‐sectional study conducted in children and adolescents aged 5 to 19 years of age, that were household contacts of parents and other relatives who were infected by SARS‐CoV‐2 in the city of Goiânia, Central Brazil, from March to October 2020. Sociodemographic and clinical data were collected from all participants. Nasopharyngeal and oropharyngeal swabs were collected and tested for SARS‐CoV‐2 RNA using real‐time reverse transcription polymerase chain reaction (RT‐PCR). Factors associated with SARS‐CoV‐2 infection and SAR were analyzed using Poisson regression. Results A total of 267 children and adolescents were investigated. The prevalence of SARS‐CoV‐2 RNA by the real‐time RT‐PCR test and/or the presence of COVID‐19 associated symptoms (anosmia/ageusia and flu syndrome) was 25.1% (95.0% Confidence Interval [95.0% CI] = 20.3‐30.6). More than half (55.1%) of the participants had sygns and symptoms. The most prevalent signs and symptoms in positive individuals were nasal congestion (62.7%), headache (55.2%), cough (50.8%), myalgia (47.8%), runny nose (47.8%), and anosmia (47.8%). The Poisson model showed that the following signs or symptoms were associated with SARS‐CoV‐2 infection: fever, nasal congestion, decreased appetite, nausea, anosmia, and ageusia. Families that had more than one infected adult, in addition to the index case, presented greater transmissibility to children and adolescents. Conclusions Our results contribute to the hypothesis that children and adolescents are not important sources of transmission of SARS‐CoV‐2 in the home environment during a period of social distancing and school closure; even though they are susceptible to infection in the household (around ¼ of our study population).

Polymorphism in metabolizing enzymes can influence drug response as well as the risk for adverse drug reactions. Nevertheless, there are still few studies analyzing the consequence of polymorphisms for the Glutathione-S-transferases (GST) gene to drug response in chronic myeloid leukemia (CML). This study reports, the influence of GSTP1*B and GSTT1/GSTM1null polymorphisms in response to imatinib in CML patients in a Brazilian population. One hundred thirty-nine CML patients from the Clinical Hospital of Goiânia, Goias, Brazil, treated with imatinib were enrolled in this study. Genotyping of GSTT1 and GSTM1 genes deletions were performed by qPCR and of GSTP1 gene was performed by RFLP-PCR. The frequency of GSTP1*1B, GSTT1 and GSTM1null polymorphisms were determined for all patients. The influence of each patient's genotypes was analyzed with the patient's response to imatinib treatment. Brazilian CML patients revealed GSTT1 and GSTM1 genes deletions. GSTT1 deletion was found in 19.3% of patients and GSTM1 deletion in 48.7% of patients with CML. GSTT1/GSTM1 deletion was found in 11.7% in Brazilian CML patients. The "G allele" of GSTP1*B, is associated with later cytogenetic response in imatinib therapy. While, the gene presence combined with GG genotype (GSTM1 present/GSTPI-GG) conferred a tend to a later cytogenetic response to patients. GSTP1*B and GSTT1/GSTM1null polymorphisms influence treatment response in CML. Brazilian CML patients presenting GSTP1 AA/AG genotypes alone and in combination with GSTT1 null reach the cytogenetic response faster, while patients presenting GSTP1-GG and GSTMI positive genotypes may take longer to achieve cytogenetic response. As a result, it allows a better prognosis, with the use of an alternative therapy, other than reducing treatment cost.

SARS-CoV-2 currently represents a serious global public health problem. Non-pharmaceutical intervention measures (NPIs) have been widely adopted, and the testing strategy since the beginning of the infection is the most effective tool for tracking, isolating, and minimizing transmission. The high operating costs and the need for sophisticated instrumentation related to gold standard diagnostic for COVID-19, Reverse Transcription quantitative Polymerase Chain Reaction (RT-qPCR), have highlighted the urgency and importance of developing and applying new diagnostic techniques, especially in places with scarce resources. Thus, alternative molecular tests, such as Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP), based on isothermal amplification have been used to detect SARS-CoV-2 using different protocols. The potential for field application of RT-LAMP is due to the lower cost and time and not requiring high-cost instrumentation. Here, we evaluate the colorimetric RT-LAMP to detect SARS-CoV-2 in a hospital environment and correlate its performance with tests performed in a reference laboratory. The analysis performed at the hospital showed high sensitivity (88.89%), specificity (98.55%), accuracy (95.83%), and a Cohen's kappa of 0.895. However, we achieved 100% of agreement when comparing the RT-LAMP results with the gold standard (qRT-PCR) results for samples with Ct < 30 in the hospital-based test. In addition, a similar performance was found in the field compared to the reference laboratory, corroborating the proposal to apply the test directly at point-of-care.

Made available in DSpace on 2020-12-12T01:17:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-03-02 Delivery efficiencies of theranostic nanoparticles (NPs) based on passive tumor targeting strongly depend either on their blood circulation time or on appropriate modulations of the tumor microenvironment. Therefore, predicting the NP delivery efficiency before and after a tumor microenvironment modulation is highly desirable. Here, we present a new erythrocyte membrane-camouflaged magnetofluorescent nanocarrier (MMFn) with long blood circulation time (92 h) and high delivery efficiency (10% ID for Ehrlich murine tumor model). MMFns owe their magnetic and fluorescent properties to the incorporation of manganese ferrite nanoparticles (MnFe2O4 NPs) and IR-780 (a lipophilic indocyanine fluorescent dye), respectively, to their erythrocyte membrane-derived camouflage. MMFn composition, morphology, and size, as well as optical absorption, zeta potential, and fluorescent, magnetic, and magnetothermal properties, are thoroughly examined in vitro. We then present an analytical pharmacokinetic (PK) model capable of predicting the delivery efficiency (DE) and the time of peak tumor uptake (tmax), as well as changes in DE and tmax due to modulations of the tumor microenvironment, for potentially any nanocarrier. Experimental PK data sets (blood and tumor amounts of MMFns) are simultaneously fit to the model equations using the PK modeling software Monolix. We then validate our model analytical solutions with the numerical solutions provided by Monolix. We also demonstrate how our a priori nonmechanistic model for passive targeting relates to a previously reported mechanistic model for active targeting. All in vivo PK studies, as well as in vivo and ex vivo biodistribution studies, were conducted using two noninvasive techniques, namely, fluorescence molecular tomography (FMT) and alternating current biosusceptometry (ACB). Finally, histopathology corroborates our PK and biodistribution results. Physics Institute Federal University of Goiás Biomagnetism Lab Physics and Biophysics Department São Paulo State University Laboratory of Pharmaceutical Nanotechnology and Drug Delivery Systems School of Pharmacy Federal University of Goiás Biological Sciences Institute Federal University of Goiás Biomagnetism Lab Physics and Biophysics Department São Paulo State University

Objective: We investigate the prevalence of TP53 germline pathogenic variants in a cohort of 83 breast cancer patients and 217 family members from the Midwest Brazilian region. Methods: All patients met the clinical criteria for hereditary breast and ovarian cancer syndrome (HBOC) and were negative for BRCA1 and BRCA2 mutations. Moreover, 40 index patients fulfilled HBOC and the Li-Fraumeni-like syndromes (LFL) criteria. The samples were tested using next-generation sequencing for TP53. Results: Three patients harbored TP53 missense pathogenic variants (p.Arg248Gln, p.Arg337His, and p.Arg337Cys), confirmed by Sanger sequencing. One patient showed a large TP53 deletion (exons 2–11), which was also confirmed. The p.R337H variant was detected in only one patient. Conclusion: This study concluded that 4 out of 83 HBOC and LFL patients presented TP53 pathogenic variants at a young age. In contrast to other Brazilian regions, the TP53 p.R337H variant appeared with low prevalence.

For some cancer subtypes, such as triple-negative breast cancer, there are no specific therapies, which leads to a poor prognosis associated with invasion and metastases. Ruthenium complexes have been developed to act in all steps of tumor growth and its progression. In this study, we investigated the effects of Ruthenium (II) complexes coupled to the amino acids methionine (RuMet) and tryptophan (RuTrp) on the induction of cell death, clonogenic survival ability, inhibition of angiogenesis, and migration of MDA-MB-231 cells (human triple-negative breast cancer). The study also demonstrated that the RuMet and RuTrp complexes induce cell cycle blockage and apoptosis of MDA-MB-231 cells, as evidenced by an increase in the number of Annexin V-positive cells, p53 phosphorylation, caspase 3 activation, and poly(ADP-ribose) polymerase cleavage. Moreover, morphological changes and loss of mitochondrial membrane potential were detected. The RuMet and RuTrp complexes induced DNA damage probably due to reactive oxygen species production related to mitochondrial membrane depolarization. Therefore, the RuMet and RuTrp complexes acted directly on breast tumor cells, leading to cell death and inhibiting their metastatic potential; this reveals the potential therapeutic action of these drugs.

The coronavirus pandemic has been causing damage to many nations, as public and private health systems deteriorate by the increasing demand. Some infected patients have culturable severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) even though not presenting any symptoms, and therefore, are probably able to transmit it. Correctly diagnosing and isolating infected patients is an important step towards preventing new infections. Current diagnostic methods rely mainly on reverse transcription quantitative polymerase chain reaction (RT-qPCR). Methods such as reverse transcription loop-mediated isothermal amplification (RT-LAMP) have risen as viable alternatives, as they are cheaper and require less infrastructure, they have the potential to be applied in low-resource scenarios and even at point-of-care. Here we report a colorimetric RT‑LAMP assay capable of detecting SARS-CoV-2 in ribonucleic acid (RNA) from saliva. In some cases, the test was able to detect viral RNA before symptom onset and even in a self-reported asymptomatic carrier. It had a limit of detection of 300 copies per reaction and showed a sensitivity of 80%, a specificity of 100%, a general accuracy of 99.59%, and a Cohen’s kappa of 0.887. The possibility of detecting positive cases even before the clinical manifestation shows great potential and can contribute to controlling the pandemic.

Ruthenium is attracting considerable interest as the basis for new compounds to treat diseases, and studies have shown that complexes with different structures have significant antineoplastic and antimetastatic potential against several types of tumors, including tumors resistant to cisplatin drugs. We examined the cytotoxic, genotoxic, and pro-apoptotic activities of six ruthenium complexes containing amino acid with general formulation [Ru(AA)(bipy)(dppb)]PF6, where AA = amino acid (alanine, glycine, leucine, lysine, methionine, or tryptophan); bipy = 2,2´-bipyridine; and dppb = [1,4-bis(diphenylphosphine)butane], against A549 (lung carcinoma) and K562 (chronic myelogenous leukemia) cancer cells. The results show that the ruthenium complexes tested were able to induce cytotoxicity in A549 and K562 cancer cells. Complex 1 containing alanine inhibited the cell viability of A549 and K562 tumor cells by inducing apoptosis, as evidenced by an increased number of Annexin V-positive cells and the induction of DNA damage and cell cycle arrest. Complex 1 was able to induce caspase-mediated apoptosis in K562 cells through the mitochondrial dysfunction, the upregulation of apoptotic genes, and the downregulation of Bcl2 anti-apoptotic gene. Besides being cytotoxic to K562 and A549 cells, ruthenium complex containing alanine shows low cytotoxicity and genotoxicity against non-tumor cells. These results suggest that the ruthenium (II) complex is a potential safe and efficient antineoplastic candidate for leukemia treatment.

The present study reports the synthesis and crystal structures of Cu(II) complexes with pyridoxal S-allyldithiocarbazate (H2L1) and pyridoxal thiosemicarbazones (H2L2 = pyridoxal-N4-phenyl-3-thiosemicarbazone and H2L3 = pyridoxal-N4-semicarbazone). The single-crystal X-ray study reveals that in all cases, the Schiff base coordinated tridentately through the ONS-donor atoms, resulting in distorted square planar coordination geometries with the copper atoms. The Cu(II) complexes with pyridoxal dithiocarbazate, [Cu(HL1)Cl]·H2O and [Cu(HL1)Br]·H2O, as well as three complexes with pyridoxal thiosemicarbazone, [Cu(HL2)Cl]·dmf, [Cu(HL2)Br]·H2O·dmf and [Cu(H2L3)Br]Br·H2O, were also characterized by spectroscopic and physical–chemical analyses. The cytotoxicity of the complexes toward two kinds of cancerous cells (Ehrlich and S-180 cells) was evaluated by an MTT assay. The complex [Cu(H2L3)Br]Br·H2O was selected to study both the cellular and molecular mechanisms underlying its promising cytotoxicity. The Hoechst 33342/PI dual-staining assay showed the typical apoptotic morphology of cancer cells, and the RT-qPCR analysis revealed that the expressions of Bax, Casp3, Casp8, Casp9 and TP53 were markedly increased in both the Ehrlich and S-180 cells exposed to 10 μM for 3 h. According to our results, this complex induces cell death through apoptosis, showing potential as a future drug against cancer. Five stable copper(II) complexes with pyridoxal have been synthesized and their crystal structure has been studied. Spectroscopy experiments and cytotoxic assays against S-180 and Ehrlich cancer cell lines were performed and showed promising results as novel antitumor drugs.

This study describes the synthesis and structure of (1E,4E)-1-(3-chlorophenyl)-5-(2,6,6-trimethylcyclohex-1-en-1-yl)penta-1,4-dien-3-one (BC I). X-ray single-crystal diffraction and Hirshfeld surface analysis describe supramolecular arrangement and topological analysis. Theoretical calculations, such as QTAIM, frontier molecular orbital, MEP and infrared spectra assignments, were performed at the B3LYP/6-311++G(d,p) level of theory. Also, this work evaluates molecular docking against DNA (PDB ID: 1BNA) and cytotoxic activity against two tumor cell lines. The BC I molecule has a half chair conformation of the cyclohexene ring, and the supramolecular arrangements are stabilized by C6–H6⋯O1 and C7–H7⋯O1 interactions. MEP and docking analyses indicate an electrophilic attack that is likely to occur on the carbonyl group.

Mayaro virus (MAYV) has historically been associated with sylvatic transmission; however, urban outbreaks have been reported in Brazil, including cases of co-detection with dengue virus (DENV). Therefore, we performed a molecular survey to investigate MAYV circulation and cocirculation with DENV within Goiania, a major city in Central-West Brazil. Among 375 subjects with arbovirus-like symptoms, 259 were positive for DENV and 26 for MAYV. Of these, 17 were coinfected with DENV-2, suggesting co-transmission of the viruses. The most common complaints at the time of inclusion were myalgia, headache, fever, arthralgia, retro-orbital pain, and skin rash. No specific symptoms were associated with MAYV when either detected alone or co-detected with DENV, compared to that when DENV was detected alone. Most MAYV-infected subjects were women with no recent travel history to rural/sylvatic areas. Phylogenetic reconstruction indicated that the MAYV identified in this study is closely related with a lineage observed in Peru, belonging to genotype D. Our results corroborate the growing circulation of MAYV in urban environments in Brazil and reinforce the need to implement laboratory diagnosis in the Unified Health System, considering that the clinical manifestations of Mayaro fever are similar to those of other arboviruses, particularly dengue. Furthermore, most cases occurred in association with DENV-2. Further phylogenetic studies are needed to evaluate MAYV, which has not been widely examined.

IR-780 iodide is a fluorescent dye with optical properties in the near-infrared region that has applications in tumor detection and photothermal/photodynamic therapy. This multifunctional effect led to the development of theranostic nanoparticles with both IR-780 and chemotherapeutic drugs such as docetaxel, doxorubicin, and lonidamine. In this work, we developed two albumin-based nanoparticles containing near-infrared IR-780 iodide multifunctional dyes, one of them possessing a magnetic core. Molecular docking with AutoDock Vina studies showed that IR-780 binds to bovine serum albumin (BSA) with greater stability at a higher temperature, allowing the protein binding pocket to better fit this dye. The theoretical analysis corroborates the experimental protocols, where an enhancement of IR-780 was found coupled to BSA at 60 °C, even 30 days after preparation, in comparison to 30 °C. In vitro assays monitoring the viability of Ehrlich ascites carcinoma cells revealed the importance of the inorganic magnetic core on the nanocarrier photothermal-cytotoxic effect. Fluorescence molecular tomography measurements of Ehrlich tumor-bearing Swiss mice revealed the biodistribution of the nanocarriers, with marked accumulation in the tumor tissue (≈3% ID). The histopathological analysis demonstrated strong increase in tumoral necrosis areas after 24 and 72 h after treatment, indicating tumor regression. Tumor regression analysis of nonirradiated animals indicate a IR-780 dose-dependent antitumoral effect with survival rates higher than 70% (animals monitored up to 600 days). Furthermore, an in vivo photothermal therapy procedure was performed and tumor regression was also verified. These results show a novel insight for the biomedical application of IR-780-albumin-based nanocarriers, namely cancer therapy, not only by photoinduced therapy but also by a nonirradiation mechanism. Safety studies (acute oral toxicity, cardiovascular evaluation, and histopathological analysis) suggest potential for clinical translation.

Infection caused by the new coronavirus (SARS-CoV-2) has become a serious worldwide public health problem, and one of the most important strategies for its control is mass testing. Loop-mediated isothermal amplification (LAMP) has emerged as an important alternative to simplify the diagnostics of infectious diseases. In addition, an advantage of LAMP is that it allows for easy reading of the final result through visual detection. However, this step must be performed with caution to avoid contamination and false-positive results. LAMP performed on microfluidic platforms can minimize false-positive results, in addition to having potential for point-of-care applications. Here, we describe a polystyrene-toner (PS-T) centrifugal microfluidic device manually controlled by a fidget spinner for molecular diagnosis of COVID-19 by RT-LAMP, with integrated and automated colorimetric detection. The amplification was carried out in a microchamber with 5 μL capacity, and the reaction was thermally controlled with a thermoblock at 72 °C for 10 min. At the end of the incubation time, the detection of amplified RT-LAMP fragments was performed directly on the chip by automated visual detection. Our results demonstrate that it is possible to detect COVID-19 in reactions initiated with approximately 10-3 copies of SARS-CoV-2 RNA. Clinical samples were tested using our RT-LAMP protocol as well as by conventional RT-qPCR, demonstrating comparable performance to the CDC SARS-CoV-2 RT-qPCR assay. The methodology described in this study represents a simple, rapid, and accurate method for rapid molecular diagnostics of COVID-19 in a disposable microdevice, ideal for point-of-care testing (POCT) systems.

Patients with pathogenic variants (PV) in the BRCA1 and BRCA2 genes have hereditary breast and ovarian cancer syndrome (HBOC). Some patients with HBOC have a family history (FH) of different types of cancer not related to the syndrome. The objective of this study was to observe the FH profile of cancer in patients with HBOC syndrome. A total of 123 patients treated at the Advanced Breast Diagnostic Center (CORA) with clinical criteria suggestive of HBOC syndrome were selected according to the National Comprehensive Cancer Network (NCCN). The collection of 4 ml of blood was performed, which was subjected to DNA extraction and PV analysis in the BRCA1 and BRCA2 genes by next generation sequencing. The data were analyzed using the Sophia DDM and Ion Reporter software. The variants were considered to be pathogenic according to the ACMG criteria. It was found that among 123 patients analyzed, 19 had HBOC syndrome, of whom 5 were related. Thus, we had 16 families with HBOC syndrome. Among the 16 families, 14 (87.5%) had FH from cancers related to HBOC syndrome, 9 (56.25%) had FH from cancers not related to HBOC syndrome, and 1 (16.25%) did not have FH cancer. A total of 8 (50%) of families with HBOC also met the NCCN criteria for other hereditary cancer syndromes, 3 (18.75%) for Li–Fraumeni syndrome (LFS) and HBOC, 3 (18.75%) for Lynch syndrome (LS) and HBOC, and 2 (12.5%) for HBOC, LFS, and LS. The most common cancers observed outside the common spectrum of HBOC syndrome in families were stomach cancer (25%), intestine (18.75%), liver (18.75%), and skin (18.75%). These data suggest the importance of a complete assessment of FH in patients with HBOC syndrome to better understand its relationship with the predisposition to different types of cancer.

The Arboviral diseases are caused by arthropod-borne viruses, such as Mayaro virus (MAYV), the etiological agent of Mayaro fever. This disease has been drawing the attention of the public health authorities for the increased number of cases likely due to virus adaptation for survival to urban areas as well as infection and multiplication in other vectors insects. Therefore, this work aimed to identify the MAYV infecting Aedes aegypti mosquitoes in Goiânia, the capital of state of Goiás, Brazil. For the development of study, the larvae of A. aegypti were collected in Basic Health Units from different regions of Goiânia then the larvae were grown to adult mosquitoes in controlled laboratory conditions. The female mosquitoes were submitted to the procedure of head and body separation. The RNAs obtained from these samples were analyzed by real-time PCR for identification of arboviruses. We only detect the presence of MAVY in the mosquitoes, in this sense our findings suggest that A. aegypti harbor MAYV in different anatomical sites, and potentially the process of vertical transmission of MAYV can occur in this vector.

Made available in DSpace on 2020-12-10T17:36:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-04-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Antimetastatic activity, high selectivity and cytotoxicity for human tumor cell lines make ruthenium(ii) complexes attractive for the development of new chemotherapeutic agents for cancer treatment. In this study, cytotoxic activities and the possible mechanism of cell death induced by three ruthenium complexes were evaluated, [Ru(MIm)(bipy)(dppf)]PF6(1), [RuCl(Im)(bipy)(dppf)]PF6(2) and [Ru(tzdt)(bipy)(dppf)]PF6(3). The results showed high cytotoxicity and selectivity indexes for the human triple-negative breast tumor cell line (MDA-MB-231) with IC(50)value and selectivity index for complex1(IC50= 0.33 +/- 0.03 mu M, SI = 4.48), complex2(IC50= 0.80 +/- 0.06 mu M, SI = 2.31) and complex3(IC50= 0.48 +/- 0.02 mu M, SI = 3.87). The mechanism of cell death induced in MDA-MB-231 cells, after treatment with complexes1-3, indicated apoptosis of the cells as a consequence of the increase in the percentage of cells in the Sub-G1 phase in the cell cycle analysis, characteristic morphological changes and the presence of apoptotic cells labeled with Annexin-V. Multiple targets of action were identified for complexes1and3with an induction of DNA damage in cells treated with complexes1and3, mitochondrial depolarization with a reduction in mitochondrial membrane potential, an increase in reactive oxygen species levels and increased expression levels of caspase 3 and p53. In addition, antimetastatic activities for complexes1and3were observed by inhibition of cell migration by the wound healing assay and Boyden chamber assay, as well as inhibition of angiogenesis caused by MDA-MB-231 tumor cells in the CAM model. Univ Fed Sao Carlos, Dept Chem, CP 676, BR-13565905 Sao Carlos, SP, Brazil Univ Fed Goias, Inst Biol Sci, Dept Genet, BR-74690900 Goiania, Go, Brazil Fed Inst Educ Sci & Technol Goias, Dept Chem, BR-74055110 Goiania, Go, Brazil Pontifical Catholic Univ Goias, Lab Expt & Biotechnol Res, Masters Program Environm Sci & Hlth, Sch Med Sci,Pharmaceut & Biomed Lab, BR-74605010 Goiania, Go, Brazil Sao Paulo State Univ, Ctr Environm Studies, BR-13506900 Rio Claro, SP, Brazil Univ Fed Goias, Inst Biol Sci, Dept Morphol, BR-74690900 Goiania, Go, Brazil Sao Paulo State Univ, Ctr Environm Studies, BR-13506900 Rio Claro, SP, Brazil CAPES: 001 FAPESP: 2016/16312-0

About 10‒15% of breast cancer cases are due to deleterious germ changes, more than half of which are located in the BRCA1 or BRCA2 genes. The screening of variants in these genes for patients at risk brings benefits for better clinical management of the patient as well as for the prevention of disease recurrences both in the proband and in their relatives. The profile of genetic variants is little known to the Brazilian population and, to date, there are no published data for the population of the central region of the country. This study aimed to analyze the profile of pathogenic variants (VP) and of uncertain significance (VUS) in the BRCA1 and BRCA2 genes in this population. We selected 102 patients seen at Hospital das Clínicas, Universidade Federal de Goiás, with clinical diagnosis of invasive ductal carcinoma that met the criteria of the National Comprehensive Cancer Networking 2016/2 for hereditary breast and ovarian cancer syndrome. A collection of 4mL of peripheral blood was carried out and submitted to subsequent extraction of genomic DNA, carried out using the PureLink kit (Invitrogen). The genes in question had all their exon-intron regions sequenced using the MiSeq device (Illumina) and the raw data were evaluated using the Sophia DDM software. The risk of in silico pathogenicity of the VUS was analyzed by the software POLYPHEN2, MutationTaster, SIFT, ESP5400, G1000, GnomAD. Of the total of 102 patients evaluated in this study, 22 (21.56%) had PV or VUS in the BRCA1 or BRCA2 genes. Of these, 16 patients (72.81%) had pathogenic variants, eight with PV in BRCA1 (c.5266dupC (2), c.3331_3334delCAAC, c.211A>G, c.3228_3229delAG, c.3700_3704delGTAAA, c.4484G>T and c.5305_5306ins20) and eight with PV in BRCA2 (c.156_157insAlu, c.4829_4830delTG, c.8488-1G>A, c.6405_6409delCTTAA (3), c.517-1G>A, c.2808_2811delACAA). A total of 7 patients (31.8%) presented VUS in these genes, one in the BRCA1 gene (c.179A>G) and five in the BRCA2 gene (c.280C>T, c.811G>A, 1096T>G, 1441A>G and c.5270A>G) of which only the variant c.1441A>G had low pathogenic potential in silico. A new PV still not described in the literature was also identified, variant c.5305_5306ins20 in the BRCA1 gene. These data suggest that all patients at risk in this population should be evaluated for PV or VUS in the BRCA1 and BRCA2 genes since the presence of these variants can help in the patient's prognosis and disease prevention.

Introdução: O atual surto de COVID-19 tornou-se uma séria ameaça à saúde das pessoas em todo o mundo. SARS-CoV-2 é facilmente transmitido através do contato com aerossóis, gotículas e fômites de pessoas infectadas. No entanto, SARS-CoV-2 também foi detectado em amostras de fezes, sugerindo uma possível contaminação fecal oral. A epidemiologia baseada em águas residuárias é uma ferramenta de monitoramento do perfil microbiológico de uma comunidade que pode contribuir para a identificação de patógenos, auxiliando na tomada de decisões, previamente aos surtos epidemiológicos. Objetivo: Este estudo monitorou a presença de SARS-CoV-2 em águas residuárias no município de Goiânia, Goiás, Brasil; estimou a prevalência da infecção correlacionando com os dados de COVID-19 clinicamente confirmados; desenvolveu um aplicativo baseado em linguagem Python para auxiliar na tomada de decisões. Métodos: Amostras de esgoto afluente e efluente foram coletadas durante os meses de janeiro a agosto de 2021 na estação de tratamento de águas residuais Dr. Hélio Seixo de Britto. A concentração viral foi realizada usando polietilenoglicol. O RNA viral foi extraído empregando o kit MagMAX’. A detecção das regiões N1 e N2 do RNA viral foi realizada por RT-qPCR. A prevalência de infecção foi estimada através da equação: NPI=CGFVαβε, a qual foi utilizada no desenvolvimento de um aplicativo para predição de números de infectados a partir da carga viral quantificada nas amostras de esgoto usando a linguagem Python. Resultados: Das 55 amostras coletadas 24 (43,63%) foram positivas, sendo 13 amostras de esgoto afluente e 15 amostras de esgoto efluente. Isto demonstra que SARS-CoV-2 não é totalmente eliminado mesmo após o tratamento. 16 amostras foram positivas para N1 (8 afluentes e 8 efluentes) e 14 amostras foram positivas para N2 (6 afluentes e 8 efluentes). Comparando o número de infectados preditos em nosso estudo com os casos relatados pelo órgão de vigilância, observa-se que a identificação de SARS-CoV-2 em esgoto consegue melhor dimensionar a dinâmica de infecção. Conclusão: A presença do RNA SARS-CoV-2 em esgotos confirma a potencialidade da vigilância ambiental como ferramenta de monitoramento, complementando a vigilância clínica.

Introdução: São conhecidos como arbovírus, os patógenos capazes de infectar vertebrados e invertebrados, através da picada de um vetor artrópode (Casseb et al., 2013). Entre os anos de 1961 até 2019, no américa latina a estimativa é que ocorreram cerca de mais de 500 mil casos de febre de Oropouche, sendo que 99,76% dos casos ocorreram no Brasil. O vírus Oropouche pertencente à família Bunyaviridae é transmitido pelo mosquito Culicoides paraensis (de Melo, 2020). O primeiro isolamento no Brasil ocorreu em 1960 do sangue de uma preguiça (Bradypus tridactylus) (Azevedo et al., 2007). Devido à grande semelhança do VORO com outras arboviroses como a Dengue, as infecções hemorrágicas são muito similares, o que dificulta o diagnóstico nos hospitais, o que afeta a notificação epidemiológica (Mor, 2021). Em consequência do grande número de subnotificações, ainda não há abordagens preventivas especificas contra o vírus (Pinheiro et al., 1982). O objetivo desse artigo foi rastrear a presença do vírus Oropouche no estado de Goiás, e mapear as regiões mais afetadas. Metodologia: Trata-se de um estudo laboratorial com pacientes do CAIS Jardim Novo Mundo em Goiânia, Goiás, que apresentavam sintomas característicos à infecção por arbovírus. Para a pesquisa, foram feitas coletas de soro de 155 pacientes de 2017 a 2020. Das 155 amostras, 79 foram submetidas a extração do RNA viral, utilizando o kit MagMAX Viral/Pathogen Nucleic Acid Isolation seguindo as orientações do fabricante. Depois de obter o RNA viral foi realizado a RT-qPCR. As sequências dos oligonucleotídeos e sondas para identificação do vírus Oropouche foram adquiridas pela empresa IDT (Integrated DNA Technologies, Coralville, IA, USA). Resultados: O presente estudo está em andamento, foram realizadas as extrações de RNA de 79 amostras de soro de pacientes com suspeita de infecção por arboviroses, após a extração foi realizada a RT-qPCR. O resultado da PCR mostrou amplificação em todas as amostras utilizando o controle endógeno RNAse P humana, porém nenhuma amostra positiva para o vírus Oropouche foi identificada até o momento. Conclusão: O rastreio do vírus na região central do Brasil possibilita a investigação dos genótipos circulantes, e a divulgação desses dados na literatura será de grande impacto devido a sua baixa notificação no país e pelo fato de que a febre de Oropouche é a segunda doença febril mais incidente no Brasil.

Thirteen new ruthenium amino acid complexes were synthesized and characterized. They were obtained by the reaction of α-amino acids (AA) with [RuCl2(P-P)(N-N)], where P-P=1,4-bis(diphenylphosphino)butane (dppb) or 1,3-bis(diphenylphosphino)propane (dppp) and N-N=4,4'-dimethyl-2,2'-bipyridine (4'-Mebipy), 5,5'-dimethyl-2,2'-bipyridine (5'-Mebipy) or 4,4'-Methoxy-2-2'-bipyridine (4'-MeObipy). This afforded a family of complexes formulated as [Ru(AA-H)(P-P)(N-N)]PF6, where AA=glycine (Gly), L-alanine (Ala), L-valine (Val), L-tyrosine (Tyr), L-tryptophan (Trp), L-histidine (His) and L-methionine (Met). All compounds were characterized by elemental analysis, spectroscopic and electrochemical techniques. The [Ru(AA-H)(P-P)(N-N)]PF6 complexes are octahedral (the AA-H ligand binding involves N-amine and O-carboxylate), diamagnetic (low-spin d6, S=0) and present bands due to electronic transitions in the visible region. 1H, 13C{1H} and 31P{1H} NMR spectra of the complexes indicate the presence of C2 symmetry, and the identification of diastereoisomers. In vitro cytotoxicity assays of the compounds and cisplatin were carried out using MDA-MB-231 (human breast) tumor cell line and a non-tumor breast cell line (MCF-10A). Most complexes present promising results with IC50 values comparable with the reference drug cisplatin and high selectivity indexes were found for the complexes containing L-Trp. The binding of two Ru-precursors of the type [RuCl2(dppb)(NN)] (N-N=4'-MeObipy or 4'-Mebipy) to the blood transporter protein human serum albumin (HSA) was evaluated by fluorescence and circular dichroism spectroscopy. Both complexes bind HSA, probably in the hydrophobic pocket near Trp214, and the Ru-complex containing 4'-MeObipy shows higher affinity for HSA than the 4'-Mebipy one.

Azathioprine (AZA) is the main drug used in immunomodulatory therapy in post-transplant patients or with autoimmune diseases. However, no study has evaluated the AZA angiogenic response. Therefore, this study investigated the effects of AZA on the angiogenic process through macroscopic, histological, and immunohistochemical analyses in chick embryo chorioallantoic membrane (CAM). Our results showed potent anti-angiogenic activity of AZA at the higher concentrations tested in the CAM assay. The histological analysis of CAM confirmed this effect, since AZA induced a significant reduction in all parameters evaluated. In addition, immunohistochemical evaluation of CAM revealed that AZA decreased TGF-β and VEGF levels, important cytokines involved in the angiogenic process. Therefore, the AZA anti-angiogenic effect identified in our study provides new information for the possible application of this drug in anticancer treatment.

Ruthenium(II)/benzonitrile complexes have demonstrated promising anticancer properties. Considering that there are no specific therapies for treating sarcoma, we decided to evaluate the cytotoxic, genotoxic, and lethal effects of cis-[RuCl(BzCN)(phen)(dppb)]PF6 (BzCN = benzonitrile; phen = 1,10-phenanthroline; dppb = 1,4-bis-(diphenylphosphino)butane), as well as the mechanism of cell death induction that occurs against murine sarcoma-180 tumor. Thus, MTT assay was applied to assess the ruthenium cytotoxicity, showing that the compound is a more potent inhibitor for the sarcoma-180 tumor cell viability than normal cells (lymphocytes). The comet assay indicated low genotoxic for normal cells. cis-[RuCl(BzCN)(phen)(dppb)]PF6 also showed moderate lethality in Artemia salina. The complex induced cell cycle arrest in the G0/G1 phase in sarcoma-180 cells. In addition, the complex caused S180 cells to die by apoptosis by an increase in Annexin-V-positive cells and morphological changes typical of apoptotic cells. Additionally, cis-[RuCl(BzCN)(phen)(dppb)]PF6 increased the gene expression of Bax, Casp3, and Tp53 in S180 cells. By using a western blot, we observed an increased protein level of TNF-R2, Bax, and p21. In conclusion, cis-[RuCl(BzCN)(phen)(dppb)]PF6 is active and selective for sarcoma-180 cells, leading to cell cycle arrest at the G0/G1 and cell death through a caspases-mediated and Tp53/p21-mediated pathway.

In this paper, four new ruthenium complexes, [Ru(N-S)(dppm)2]PF6 (1), [Ru(N-S)(dppe)2]PF6 (2), [Ru(N-S)2(dppp)] (3) and [Ru(N-S)2(PPh3)2] (4) [dppm = 1,1-bis(diphenylphosphino)methane, dppe = 1,2-bis(diphenylphosphino)ethane, dppp = 1,3-bis(diphenylphosphino)propane, PPh3 = triphenylphosphine and N-S = 2-mercaptopyrimidine anion] were synthesized and characterized using spectroscopy techniques, molar conductance, elemental analysis, electrochemical techniques and X-ray diffraction. The DNA binding studies were investigated using voltammetry and spectroscopy techniques. The results show that all complexes exhibit a weak interaction with DNA. HSA interaction with the complexes was studied using fluorescence emission spectroscopy, where the results indicate a spontaneous interaction between the species by a static quenching mechanism. The cytotoxicity of the complexes was evaluated against A549, MDA-MB-231 and HaCat cells by MTT assay. Complexes (1) and (2), which are very active against triple negative MDA-MB-231, were subjected to further biological tests with this cell line. The cytotoxic activity triggered by the complexes was confirmed by clonogenic assay. Cell cycle analyses demonstrated marked anti-proliferative effects, especially at the G0/G1 and S phases. The morphological detection of apoptosis and necrosis - HO/PI and Annexin V-FITC/PI assay, elucidated that the type of cell death triggered by these complexes was probably by apoptosis. The in vivo toxicological assessment performed on zebrafish embryos revealed that complexes (1) and (2) did not present embryotoxic or toxic effects during embryonic and larval development showing that they are promising new prototypes of safer and more effective drugs for triple negative breast cancer treatment.

Resumo O objetivo detse artigo é de descrever um protocolo de isolamento das células mononucleares da medula óssea de coelhos, seguido de purificação celular por depleção negativa com o anticorpo monoclonal CD45 e posterior expansão em meio de cultura MesenCult®. Dez coelhos machos adultos, da raça Nova Zelândia, com idade média de 1,0±0,2 anos e peso médio 3,5±0,24kg, foram utilizados para padronização da metodologia. O isolamento das células mononuclares da medula óssea foi realizado pelo gradiente de densidade Ficoll-paque® e a purificação e obtenção das células- pela depleção negativa com o anticorpo monoclonal CD45 em base imunomagnética. A população celular obtida foi expandida posteriormente em meio de cultura MesenCult®. No isolamento pelo gradiente de icoll-Paque® foi obtido um rendimento médio de 7,31x106 células/mL. Após purificação e obtenção das possíveis células-tronco mesenquimais pela base imunomagnética, houve um decréscimo do rendimento para 2,28x106 células/mL, mas o processo de expansão foi incrementado pelo cultivo celular. Os resultados indicaram que as células obtidas da fração mononuclear da medula óssea, cultivadas in vitro foram capazes de gerar células aderentes 24 horas após o cultivo, com predominância de células fibroblastóides sugestivas de células-tronco mesenquimais. Concluiu-se que a obtenção de células-tronco mesenquimais pode ser alcançada após purificação das células mononucleares da medula óssea de coelhos pelo método imunomagético, o meio de cultura MesenCult® proporciona um ambiente adequado para a rápida expansão in vitro e o número de passagens exerce influência negativa sobre as características morfológicas das células.

Anticancer potential of ruthenium complexes has been widely investigated, but safety evaluation studies are still scarce. Despite of ruthenium-based anticancer agents are known to cause fewer side effects compared to other metal-based drugs, these compounds are not fully free of toxicity, causing mainly nephrotoxicity. Based on the promising results from antitumor activity of the complexes [Ru(L-Met)(bipy)(dppb)]PF6 (RuMet) and [Ru(L-Trp)(bipy)(dppb)]PF6 (RuTrp), for the first time we investigated the toxicity profile of these complexes in rodent and zebrafish models. The acute oral toxicity was evaluated in Swiss mice. The mutagenic and genotoxic potential was determined by a combination of Micronucleus (MN) and Comet assay protocols, after exposure of Swiss mice to RuMet and RuTrp in therapeutic doses. Zebrafish embryos were exposed to these complexes, and their development observed up to 96 h post-fertilization. RuMet and RuTrp complexes showed low acute oral toxicity. Recorded behavioral changes were not recorded, nor were macroscopic morphological changes or structural modifications in the liver and kidneys. These complexes did not cause genetic toxicity, presenting a lack of micronuclei formation and low DNA damage induction in the cells from Swiss mice. In contradiction, cisplatin treatment exhibited high mutagenicity and genotoxicity. RuMet and RuTrp showed low toxicity in the embryo development of zebrafish. The RuMet and RuTrp complexes demonstrated low toxicity in the two study models, an interesting property in preclinical studies for novel anticancer agents.

Background. ThePPARG2Pro12Ala (rs1801282) andIL6-174G >C (rs1800795) have important function in body weight regulation and a potential role in obesity risk. We aimed to investigate the association betweenPPARG2Pro12Ala andIL6-174G >C variants and the genotypes interaction with body composition, metabolic markers, food consumption, and physical activity in severely obese patients.Methods. 150 severely obese patients (body mass index (BMI) ≥ 35 kg/m2) from Central Brazil were recruited. Body composition, metabolic parameters, physical activity, and dietary intake were measured. The genotype was determined by the qPCR TaqMan Assays System. Multiple linear regression and multiple logistic regression models were fitted adjusting for confounders.Results. Ala carriers of the Pro12Ala polymorphism had higher adiposity measures (BMI:p=0.031, and fat mass:p=0.049) and systolic blood pressure (p=0.026) compared to Pro homozygotes. We found no important associations between the -174G >C polymorphism and obesity phenotypes. When genotypes were combined, individuals with genotypes ProAla + AlaAla and GC + CC presented higher BMI (p=0.029) and higher polyunsaturated fatty acids (PUFAs) consumption (p=0.045) compared to the ones with genotypes ProPro and GG, and individuals carriers of thePPARG2Ala allele only (genotype ProAla + AlaAla and GG) had higher fat mass and systolic and diastolic blood pressure compared to the ones with genotypes ProPro and GG.Conclusions. Severely obese individuals carrying the Ala allele of thePPARG2Pro12Ala polymorphism had higher measures of adiposity and blood pressure, while no important associations were found for theIL6-174G >C polymorphism.