1 Laboratory of Diphtheria and Corynebacteria of Medical Relevance, Faculty of Medicine, University of the State of Rio de Janeiro, UERJ, Rio de Janeiro, RJ, Brazil
2 Hospital Infection Control ...Committee, National Cancer Institute, INCA, Health Ministry, Rio de Janeiro, RJ, Brazil
3 Roberto Alcantara Gomes Biology Institute, University of the State of Rio de Janeiro, UERJ, Rio de Janeiro, RJ, Brazil
Correspondence Ana L. Mattos-Guaraldi guaraldi{at}pq.cnpq.br
Received April 20, 2009
Accepted July 21, 2009
Corynebacterium diphtheriae still represents a global medical challenge, particularly due to the significant number of individuals susceptible to diphtheria and the emergence of non-toxigenic strains as the causative agents of invasive infections. In this study, we characterized the clinical and microbiological features of what we believe to be the first case of C. diphtheriae infection of a percutaneous nephrostomy catheter insertion site in an elderly patient with a fatal bladder cancer. Moreover, we demonstrated the potential role of adherence, biofilm formation and fibrin deposition traits in C. diphtheriae from the catheter-related infection. Non-toxigenic C. diphtheriae isolated from the purulent discharge (named strain BR-CAT5003748) was identified by the API Coryne system (code 1 010 324) and a multiplex PCR for detection of dtxR and tox genes. Strain BR-CAT5003748 showed resistance to oxacillin, ceftazidime and ciprofloxacin. In experiments performed in vitro , the catheter isolate was classified as moderately hydrophobic and as moderately adherent to polystyrene surfaces. Glass provided a more effective surface for biofilm formation than polystyrene. Micro-organisms adhered to (>1.5 x 10 6 c.f.u.) and multiplied on surfaces of polyurethane catheters. Microcolony formation (a hallmark of biofilm formation) and amorphous accretions were observed by scanning electron microscopy on both external and luminal catheter surfaces. Micro-organisms yielded simultaneous expression of localized adherence-like and aggregative-like (LAL/AAL) adherence patterns to HEp-2 cells. Interestingly, the coagulase tube test resulted in the formation of a thin layer of fibrin embedded in rabbit plasma by the non-toxigenic BR-CAT5003748 strain. In conclusion, C. diphtheriae should be recognized as a potential cause of catheter-related infections in at-risk populations such as elderly and cancer patients. LAL/AAL strains may be associated with virulence traits that enable C. diphtheriae to effectively produce biofilms on catheter surfaces. Biofilm formation and fibrin deposition could have contributed to the persistence of C. diphtheriae at the infected insertion site and the obstruction of the nephrostomy catheter.
These authors contributed equally to this work.
Introduction: In the clinical setting, low KMT2E expression has been associated with poor prognosis in both acute myeloid leukemia (Damm et al. J Clin Oncol. 2011 Feb 20;29(6):682-9) and acute ...promyelocytic leukemia (APL) (Lucena-Araujo et al. Br J Haematol. 2014 Aug;166(4):540-9). Member of the Trithorax-group, the KMT2E is implicated in the positive transcriptional control of a specific set of genes related to hematopoiesis and differentiation processes in many cell types. Functional analyses showed that the encoded protein is involved in terminal myeloid differentiation and facilitate retinoic-acid induced granulopoiesis in human promyelocytes. Using lentiviral gene transfer, we hypothesized that the forced expression of KMT2E is implicated in the ATRA-induced granulocytic differentiation in APL. Aims: To investigate the effects of overexpression of KMT2E gene in cell viability and signaling in response to ATRA plus anthracycline or arsenic trioxide (ATO) treatment in APL cell lines. Methods: NB4 and NB4-R2 (resistant to ATRA treatment) cell lines were transduced with pMEG or pMEG- KMT2E lentiviral vectors and cultured with stable selection using puromycin. Before all functional experiments, cells were tested for KMT2E gene and protein expression by qPCR and confocal microscopy, respectively. The cells transduced with KMT2E and control cells were evaluated for cell viability (MTT assay), proliferation (Ki-67 staining), cell cycle (flow cytometry), mitochondrial reactive oxygen species (ROS) production (flow cytometry) in basal conditions and under treatment with ATRA (1 μM), ATO (0.187, 0.375, 0.75, 1.5 and 3 μM), and the association of ATO (1 μM) plus ATRA (1 μM) or cytarabine (150 nM). The granulocytic differentiation in response to ATRA treatment was evaluated based on the CD11b expression (flow cytometry) plus morphological analysis (Leishman staining). The expression of the main target genes (p21, MDM2, and BMI1) encoding for proteins known to be involved in KMT2E function were evaluated by qPCR. Results: Overexpression of KMT2E was significantly associated with higher cell proliferation rates in basal or ATO and cytarabine treatment compared to vehicle treated cells (all P<.001). KMT2E overexpression significantly increase cell proliferation in NB4, but not in NB4-R2 cells (Ki-67 mean intensity of fluorescence (MFI) value - pMEG: 489±10 vs KMT2E : 1432±25; P<.001). The distribution of cells in the phases of the cell cycle was similar between groups. Using a nonlinear regression analysis, IC50 for ATO was of 2.96 and 6 μM for NB4 pMEG and KMT2E transduced cells, respectively. NB4- KMT2E cells showed lower levels of ROS generation upon ATO treatment (1.5 μM) compared to controls (ROS - MFI value - pMEG: 270.6±6 vs KMT2E : 84±11; P <.001). In the attempt to assess the role of KMT2E in granulocytic differentiation, NB4- KMT2E cells were treated with ATRA 1 μM. The cells containing an extra copy of KMT2E gene showed more differentiation rate in 72, 96 and 120 hours compared to empty vector transduced cells (all p<.05). Considering the expression of target genes of KMT2E, only p21 showed an increase in KMT2E cells treated with ATRA. Conclusion: In summary, the overexpression of KMT2E is associated with higher proliferation capacity and increased resistance to ATO treatment, associated with less ROS generation.
No relevant conflicts of interest to declare.
INTRODUCTION:
Acute promyelocytic leukemia (APL) cells have been shown to secrete IL-1β, TNFα and IL-6 in vitro and particularly the first one has a relevant role in the APL-associated coagulopathy. ...All-trans retinoic acid (ATRA) treatment decreased levels of IL-8 and increased of IL-1β and G-CSF secreted in the medium. However, the serial analysis of cytokine levels in the plasma of patients with APL during ATRA treatment has never been performed. Considering the importance of inflammation in the pathogenesis of APL-associated coagulopathy, we have analyzed plasma levels of IL-8, IL-1β, IL-6, IL-10, TNF and IL-12p70 in 19 Brazilian patients with APL treated with ATRA and anthracyclines according to the International Consortium on APL protocol.
METHODS:
We retrospectively analyzed the levels of pro- and anti-inflammatory cytokines in previously frozen plasma samples of 19 APL patients, enrolled in the IC-APL study. Samples were collected at diagnosis (D0) and in the 3rd (D3) and 7th (D7) days of treatment with ATRA and anthracyclines. The control group consisted of 11 healthy adult volunteers. IL-8, IL-1β, IL-6, IL-10, TNF and IL-12p70 were measured using cytometric bead array (CBA-Human Inflammatory Cytokine Kit, BD Biosciences) . The results were analyzed using FCAP Array software (BD Biosciences). The cytokine levels were correlated with white blood cell count (WBC) as well as with the frequency of death during induction (early death) and overall survival (OS). Statistical analysis was performed by using either Mann-Whitney (for comparison of two groups) or ANOVA (comparison of three or more groups) test. Fisher's exact test was used to associate cytokine levels and white blood cells (WBC) count. Overall survival analysis was considered from the date of diagnosis to date of death or last contact. Overall survival curves were prepared by the Kaplan-Meier method and compared by the log-rank test. Significance was set at p <0.05.
IL-1β, IL-10, TNF and IL-12p70 cytokines were not detected in the plasma from all APL patients and healthy controls, except for the presence of low levels of IL-10 in 3 APL patients at D0 (median concentration: 0 pg/mL; range 0 to 4.5 pg/mL). Plasma IL-8 (median 25.9 pg/mL; range 2.6 to 285.5 pg/mL) and IL-6 levels (median 5.0 pg/mL; range 0 to 580.1 pg/mL) at D0 were higher in APL patients compared with healthy controls (undetected in all samples). ATRA significantly decreased IL-8 plasma levels at D7 in 84.2% of APL patients (p<0.005, D0 vs D7) whereas the plasma concentrations of IL-6 did not change with treatment. There were no correlation between the plasma levels of IL-8 and high (WBC > 104) or low (WBC < 104) risk groups of APL patients. Furthermore, IL-8 and IL-6 levels were not associated with early mortality with IL-6 and IL-8 plasma concentrations of 0.8 vs. 6.8 and 5.1 vs. 34.1 in patients who died or not during induction. Moreover, there was no difference in the overall survival in patients with IL-6 levels above or below the median value of the group (p=0.538; 53.2 vs. 75.2 months; 95% confidence interval = 32.9 to 73.5). The same was observed for IL-8 (p=0.59; 61.5 vs. 65.0 months; 95% confidence interval = 45.6 to 77.4).
CONCLUSIONS:
APL patients exhibit increased cytokine plasma levels of the pro-inflammatory cytokines IL-8 and IL-6 but these parameters were not associated with known prognostic factors or with disease outcome.
No relevant conflicts of interest to declare.
The supramolecular architectures of amide-containing compounds are highly dependent on the side-chain substituents, although the potential impact of isoxazole substituents on polymorph formation has ...not been thoroughly explored. Hence, three distinct forms of
N
1
,
N
3
-bis(5-methylisoxazol-3-yl)malonamide (
1
) were obtained and characterized: two polymorphic forms and one solvate. An in-depth analysis of the interactions and energy content of the crystals based on supramolecular clusters allowed us to propose crystallization mechanisms (crystal retrosynthesis). Specifically, the energy similarities between the interaction of the first sites NH
amide
O&z.dbd;C
amide
(form
1I
) and the symmetric sites NH
amide
N
isox
(form
1II
) were found to contribute to their formation. Nonetheless, the presence of DMSO resulted in the formation of form
1III
, where the solvent molecule disrupted amide-amide interactions. The first nuclei are more stable than forms
1I
and
1II
. The compound of
N
1
,
N
2
-bis(5-methylisoxazol-3-yl)oxalamide (
2
) was used as a comparison, and through the absence of polymorphs, revealed that the central carbon in molecule
1
allows a flexible adaptation that leads to the three forms. These findings suggest that variations in solvents, flexibility, and the presence/absence of amide-amide interactions can modulate the competition between amide-containing isoxazole compounds.
Variations in solvents, flexibility, and the presence/absence of amide-amide interactions can modulate the competition between amide-containing isoxazole polymorphs.
ABSTRACT This study evaluated enamel mineral content and surface microhardness before and after bleaching treatment using 10% carbamide peroxide (CP) containing calcium (Ca) or amorphous calcium ...phosphate (ACP). Thirty-six bovine slabs were randomly allocated into 3 groups (n = 12) according to bleaching treatment: G1 - Opalescence PF 10% (CP), G2 -NiteWhite ACP (CP+ACP), and G3 - Opalescence PF (10%) with calcium (CP+CA). The bleaching agent was applied on enamel surface for 6 h/day over a period of 21 days. Enamel surface was evaluated by Knoop microhardness (KNH) and micro energy-dispersive X-ray fluorescence spectrometry (p-EDXRF) at baseline and at after bleaching treatment. Data were statistically analyzed by repeated measures ANOVA and Tukey's test (a = 0.05). There was a significant decrease in microhardness after bleaching treatments for all study groups, but no difference between bleaching gels. There was no difference in the Ca/P ratio measured by p-EDXRF for all groups at the study times, but the mean value was lower in group CP+CA than in group CP+ACP. Group CP was similar to both CP+ACP and CP+CA. It can be concluded that enamel microhardness decreased after the bleaching process, regardless of the presence of calcium or ACP, but there was no significant change in the Ca/P ratio of enamel after bleaching for each tested gel. This indicates that the bleaching gels have erosive potential, causing softening of enamel without promoting surface loss, regardless of the presence of calcium of ACP ions.
Objectives: The objective of this study is to analyze the work of pharmaceutical pharmacists working in primary health care in municipalities in the state of Rio Grande do Sul. Methods: The study ...included the 18 municipalities that host the Regional Health Coordination of the Health Secretariat of the State of Rio Grande do Sul and municipalities, with more than 100 thousand inhabitants, totaling 30 municipalities. Data collection was used an electronic questionnaire distributed from March to May 2021 through electronic means. Results: We obtained 77 respondents with a mean age of 39.9 years, prevalence of white color and women. Of all participants, 58 (86.6%) had a post-graduation in Lato Sensu. In professional practice, 33 (42.9%) were pharmacists in charge of the technical and 12 (15.5%) worked in more than one function in the municipality. Regarding the working day, an average working day of 35.3 hours per week was observed. Conclusion: Asymmetry in the distribution of the pharmacist’s workforce and work in more than one function stands out. This study can help the management of pharmaceutical care in the organization and ensure the quality of the services provided, and it is suggested studies that evaluate the intensity of work of pharmacists.
Determining drought tolerance in plants is an increasingly important feature due to the reduction of water resources, since water stress is one of the main environmental factors that limit ...agricultural growth and productivity. The objective of this study was to evaluate the tolerance of crambe (Crambe abyssinica Hochst) genotypes submitted to water stress induced by polyethylene glycol during germination and early growth of seedlings. A randomized block experimental design was used in a factorial scheme consisting of five crambe genotypes (FMS Brilhante, FMS CR 1203, 1307, 1312 and 1326) and five levels of osmotic potential 0.0 (control), -0.2, -0.4, -0.5 and -0.6 MPa in five replicates of 40 seeds. Germination rate (%), normal seedling development (%), germination speed index, root and shoot length, total fresh matter, and water content of seedlings (%) were analyzed. Physiological quality of seeds and initial development of crambe genotypes was improved in the group submitted to Ψw = -0.2 MPa. Germination and vigor index of crambe seeds were hampered by reduction of the potential to -0.4 MPa. The genotype FMS CR 1203 was the most tolerant to water stress, whereas FMS CR 1307 and 1312 were the most sensitive, as corroborated by PCA.
The polynomial NARMAX (Nonlinear AutoRegressive Moving Average model with eXogenous input) is a model that represents the dynamics of physical systems. This polynomial contains information from the ...past of the inputs and outputs of the process, that is, it is a recursive model. In digital computers this generates the propagation of the rounding error. Our procedure is based on the estimation of the maximum value of the lower bound error considering an arbitrary number of pseudo-orbits produced from different natural interval extensions, and a posterior Lyapunov exponent calculation. We applied successfully our technique for two identified models of the systems: sine map and Duffing-Ueda oscillator
Objetivos: O objetivo deste estudo é analisar o trabalho dos farmacêuticos da assistência farmacêutica atuantes na atenção básica em saúde (ABS) de municípios do estado do Rio Grande do Sul. Métodos: ...Foram incluídos no estudo 18 municípios sede das Coordenadorias Regionais de Saúde da Secretaria de Saúde do Estado do RS e municípios, com mais de 100 mil habitantes, totalizando 30 municípios. Para a coleta de dados utilizou-se um questionário eletrônico distribuído no período de março a maio de 2021 através de meios eletrônicos. Resultados: Obteve-se 77 respondentes com média de idade de 39,9 anos, prevalência da cor branca e mulheres. De todos os participantes, 58 (86,6%) tinham pós-graduação na modalidade Lato Sensu. Na atuação profissional, 33 (42,9%) eram farmacêuticos responsável técnico e 12 (15,5%) atuavam em mais de uma função no município. Em relação à jornada de trabalho, observou-se uma média de jornada de trabalho de 35,3 horas semanais. Conclusão: Destaca-se assimetria na distribuição da força de trabalho do farmacêutico e laboração em mais de uma função. O presente estudo pode auxiliar a gestão da assistência farmacêutica na organização e garantir a qualidade dos serviços prestados e sugere-se estudos que avaliam a intensidade de trabalho dos farmacêuticos.
In this paper, a Propositional Temporal Logic (PTL) inference engine is implemented in the cognitive level of a Concurrent Autonomous Agent (CAA). The hardware architecture of the system is an ...embedded microcontrollers network designed to fit the concurrency requirements of the cognitive model of the aforementioned agent. The architecture of the agent comprises three levels, namely, the reactive level, instinctive level and the cognitive level, which runs concurrently. The reactive level is embedded in a PSoC 5LP, consisting of behaviours created over a embedded kinematic controller. The instinctive level runs in an ARM mbed, which receives perceptions from and sends the active behaviour to the reactive level through a CAN bus. This level uses a Knowledge Based System (KBS) with First Order Logic (FOL) as automatic reasoning mechanism. The cognitive level runs an implementation of the METATEM algorithm on a DNP/2486 which, in turn, receives symbolic information from the instinctive level to update its logical world model. It also sends new local goals to instinctive level through an Ethernet network. Finally, the architecture used in the experiments is presented, followed by the results.
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