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Citrus canker disease, caused by Xanthomonas citri subsp. citri, poses a significant threat to global citrus production. The control of the disease in the field relies mainly on the use of conventional tools such as copper compounds, which are harmful to the environment and could lead to bacterial resistance. This scenario stresses the need for new and sustainable technologies to control phytopathogens, representing a key challenge in developing studies that translate basic into applied knowledge. During infection, X. citri subsp. citri secretes a transcriptional activator-like effector that enters the nucleus of plant cells, activating the expression of the canker susceptibility gene LATERAL ORGAN BOUNDARIES 1 (LOB1). In this study, we explored the use of antisense oligonucleotides (ASOs) with phosphorothioate modifications to transiently inhibit the gene expression of CsLOB1 in Citrus sinensis. We designed and validated three potential ASO sequences, which led to a significant reduction in disease symptoms compared with the control. The selected ASO3-CsLOB1 significantly decreased the expression level of CsLOB1 when delivered through two distinct delivery methods, and the reduction of the symptoms ranged from approximately 15 to 83%. Notably, plants treated with ASO3 did not exhibit an increase in symptom development over the evaluation period. This study highlights the efficacy of ASO technology, based on short oligonucleotide chemically modified sequences, as a promising tool for controlling phytopathogens without the need for genetic transformation or plant regeneration. Our results demonstrate the potential of ASOs as a biotechnological tool for the management of citrus canker disease.

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BACKGROUND: Xylella fastidiosa is a multi-host bacterium that can be detected in hundreds of plant species including several crops. Diseases caused by X. fastidiosa are considered a threat to global food production. The primary method for managing diseases caused by X. fastidiosa involves using insecticides to control the vector. Hence, it is necessary to adopt new and sustainable disease management technologies to control not only the insect but also the bacteria and plant health. We demonstrated that N-acetylcysteine (NAC), a low-cost cysteine analogue, is a sustainable molecule that can be used in agriculture to decrease the damage caused by X. fastidiosa and improve plant health. RESULTS: Using 15N-NAC we proved that this analogue was absorbed by the roots and transported to different parts of the plant. Inside the plant, NAC reduced the bacterial population by 60-fold and the number of xylem vessels blocked by bacterial biofilms. This reflected in a recovery of 0.28-fold of the daily sap flow compared to health plants. In addition, NAC-treated citrus variegated chlorosis (CVC) plants decreased the oxidative stress by improving the activity of detoxifying enzymes. Moreover, the use of NAC in field conditions positively contributed to the increase in fruit yield of CVC-diseased plants. CONCLUSION: Our research not only advances the understanding of NAC absorption in plants, but also indicates its dual effect as an antimicrobial and antioxidant molecule. This, in turn, negatively affects bacterial survival while improving plant health by decreasing oxidative stress. Overall, the positive field-based evidence supports the viability of NAC as a sustainable agricultural application. © 2024 Society of Chemical Industry.

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Xanthomonas is an important genus of plant-associated bacteria that causes significant yield losses of economically important crops worldwide. Different approaches have assessed genetic diversity and evolutionary interrelationships among the Xanthomonas species. However, information from clustered regularly interspaced short palindromic repeats (CRISPRs) has yet to be explored. In this work, we analyzed the architecture of CRISPR-Cas loci and presented a sequence similarity-based clustering of conserved Cas proteins in different species of Xanthomonas. Although absent in many investigated genomes, Xanthomonas harbors subtype I-C and I-F CRISPR-Cas systems. The most represented species, Xanthomonas citri, presents a great diversity of genome sequences with an uneven distribution of the CRISPR-Cas systems among the subspecies/pathovars. Only X. citri subsp. citri and X. citri pv. punicae have these systems, exclusively of subtype I-C system. Moreover, the most likely targets of the X. citri CRISPR spacers are viruses (phages). At the same time, few are plasmids, indicating that CRISPR/Cas system is possibly a mechanism to control the invasion of foreign DNA. We also showed in X. citri susbp. citri that the cas genes are regulated by the diffusible signal factor, the quorum sensing (QS) signal molecule, according to cell density increases, and under environmental stress like starvation. These results suggest that the regulation of CRISPR-Cas by QS occurs to activate the gene expression only during phage infection or due to environmental stresses, avoiding a possible reduction in fitness. Although more studies are needed, CRISPR-Cas systems may have been selected in the Xanthomonas genus throughout evolution, according to the cost-benefit of protecting against biological threats and fitness maintenance in challenging conditions.

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PURPOSE: Periodontitis and coronavirus disease (COVID-19) share risk factors and activate similar immunopathological pathways, intensifying systemic inflammation. This study investigated the clinical, immunological and microbiological parameters in individuals with COVID-19 and controls, exploring whether periodontitis-driven inflammation contributes to worsening COVID-19 endpoints. METHODS: Case (positive RT-PCR for SARS-CoV-2) and control (negative RT-PCR) individuals underwent clinical and periodontal assessments. Salivary levels of TNF-α, IL-6, IL-1ß, IL-10, OPG, RANKL, neutrophil extracellular traps, and subgingival biofilm were analyzed at two timepoints. Data on COVID-19-related outcomes and comorbidity information were evaluated from medical records. RESULTS: Ninety-nine cases of COVID-19 and 182 controls were included for analysis. Periodontitis was associated with more hospitalization (p = 0.009), more days in the intensive care unit (ICU) (p = 0.042), admission to the semi-ICU (p = 0.047), and greater need for oxygen therapy (p = 0.042). After adjustment for confounders, periodontitis resulted in a 1.13-fold increase in the chance of hospitalization. Salivary IL-6 levels (p = 0.010) were increased in individuals with COVID-19 and periodontitis. Periodontitis was associated with increased RANKL and IL-1ß after COVID-19. No significant changes were observed in the bacterial loads of the periodontopathogens Porphyromona gingivalis, Aggregatibacter actinomycetemcomitans, Tanerella forsythia, and Treponema denticola. CONCLUSIONS: Periodontitis was associated with worse COVID-19 outcomes, suggesting the relevance of periodontal care to reduce the burden of overall inflammation. Understanding the crosstalk between SARS-CoV-2 infection and chronic conditions such as periodontitis that can influence disease outcome is important to potentially prevent complications of COVID-19.

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Background: To describe the indications and outcomes of BRCA mutation carriers undergoing nipple-sparing mastectomy (NSM). Methods: In this retrospective study, 76 BRCA mutation carriers with no cancer who opted to undergo risk reduction NSM or diagnosed with breast cancer (BC) who opted to undergo therapeutic NSM were included. Results: Indications for NSM: cancer treatment (n = 33), bilateral risk reduction (n = 39) and contralateral prophylactic NSM (n = 4). In a mean follow-up of 45 months (median: 30 months), one patient (2.5%) undergoing risk-reducing NSM developed a new BC. One (3%) local, one (3%) ipsilateral axillary and one (3%) distant recurrence were observed in BC patients. No partial or total nipple necrosis occurred. Conclusion: NSM is safe for reducing the risk of BC development in BRCA mutation carriers and for treating cancer.

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BACKGROUND: Diabetic retinopathy (DR) is a common complication of DM and may go unnoticed until irreversible damage occurs. Its screening can contribute to the early detection. Although, there are no studies which investigate the ability of digital retinography to detect vascular changes in pre-diabetic patients. OBJECTIVE: Identify the prevalence and severity of RD in patients with pre-diabetes. METHODS: Cross-sectionalstudy carried out in a sample of patients with pre-diabetes and weight excess characterized from January 2020 to April 2023. Sociodemographic and clinical variables were collected, in addition to lifestyle habits. Retinographic evaluation was also performed using a Digital Retinography. For the analysis of all variables, the adopted significance level was 5%. The software used for the analysis was SPSS version 25.0. RESULTS: Of 108 patients selected 7.1% have alteration in the exam indicating DR. Among the participants with diabetic retinopathy, four had the moderate form (50%), three the moderate form (37%) and only one participant had the severe form (13%). CONCLUSIONS: Our findings highlight the importance of preventive measures and adequate control of these conditions in pre-diabetic patients, in order to prevent or delay the progression of diabetic retinopathy and, consequently, reduce the risk of blindness and other ocular complications.

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Pseudomonas aeruginosa an opportunistic pathogen that causes infections in hospitals and has high morbidity and mortality rates. In addition, it is a widely distributed environmental bacterium that can colonise a variety of habitats. Although wild animals do not have access to antibiotics, antibacterial resistance in these animals has increasingly been reported worldwide. Although the presence of Klebsiella pneumoniae carbapenemase (KPC) is uncommon in P. aeruginosa, it has been increasingly reported. This study examined KPC-2-producing P. aeruginosa in wild animals. A total of 27 P. aeruginosa isolates were obtained from clinical cases treated at the Microbiology Laboratory of the Veterinary Hospital of UFMT, Brazil. P. aeruginosa and blaKPC-2 carbapenemase resistance genes were identified using PCR. Antimicrobial susceptibility of KPC-producing P. aeruginosa was evaluated using the disk diffusion method. The blaKPC-2 gene was detected in 40.7% of the isolates (11/27). The rates of antimicrobial resistance and intermediate sensitivity were as follows: piperacillin/tazobactam (44.4%), imipenem (29.6%), meropenem (51.8%), amikacin (77.8%), cefepime (85.2%), and ciprofloxacin (70.4%). Twelve isolates were classified as Multidrug-resistant (MDR). This study presents the first report of P. aeruginosa with the blaKPC-2 gene in wild animals in Brazil, highlighting the importance of molecular research on resistance genes in P. aeruginosa from a One-Health perspective.

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BACKGROUND: This study aimed to evaluate the risk and protective factors associated with anxiety and depression symptoms in cancer patients at an advanced stage of cancer. METHODS: A cross-sectional study was conducted on patients with advanced cancer who were receiving palliative care. Cancer patients aged 18 years or older, with preserved cognitive function who completed the questionnaires were eligible. The questionnaires of Hospital Anxiety and Depression Scale (HADS) and health related of quality of life questionnaire; the European Organization for Research and Treatment of Cancer (EORTC-C30) were applied. Outcome variables were the depression and anxiety symptoms of cancer patients under palliative care, according to the answers to the 14 items that make up the HADS Scale. The analysis used the R software, version 4.2.0. RESULTS: Seventy cancer patients with advanced cancer were included. The colon was the most common neoplastic diagnostic (20%), followed by breast (12.9%) and lung (10%). The prevalence of depression was 44.3%, 25.7% anxiety and 52.9% had both symptoms. Patients with a high level of functionality had a lower chance of anxiety (OR = 0.80;p = 0.025), depression (OR = 0.82; p = 0.007), and anxiety and depression (OR = 0.82p = 0.008). We observed a lower chance of depression and depression/anxiety who showed a high level of Overall Performance. Three symptoms increased the chance of depression/anxiety: nausea/vomiting (p = 0.019), fatigue (0.031), loss of appetite (0.048). CONCLUSION: This study found high prevalence of anxiety and depression.Better quality of life and functionality were negatively associated with these outcomes. Examining the patient's functions will assist the clinician in alleviating symptoms of anxiety and depression, giving cancer patients in palliative care more dignity. TRIAL REGISTRATION: Not applicable.

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BACKGROUND AND OBJECTIVES: Radical prostatectomy (RP) is a definitive surgical therapy for localized prostate cancer. Evidence suggests that the poor ergonomics of surgeons during RP may lead to work-related musculoskeletal disorders and loss of productivity. Since each surgery modality has its physical demands, we compared the ergonomic risk between laparoscopic (LRP) and robotic-assisted (RARP) radical prostatectomy. METHODS: The study assessed the posture of 10 urological surgeons during LRP and RARP surgeries with the Rapid Entire Body Assessment (REBA) scale. RESULTS: We found that the RARP approach resulted in lower REBA scores over the LRP procedure. CONCLUSIONS: Robotic surgery improves body posture for the urological surgeon like in other medical specialties. However, the surgeons display harmful postures in both surgeries.

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