Molecular insights into the dynamic modulation of bacterial ClpP function and oligomerization by peptidomimetic boronate compounds.

Bacterial caseinolytic protease P subunit (ClpP) is important and vital for cell survival and infectivity. Recent publications describe and discuss the complex structure-function relationship of ClpP and its processive activity mediated by 14 catalytic sites. Even so, there are several aspects yet to be further elucidated, such as the paradoxical allosteric modulation of ClpP by peptidomimetic boronates. These compounds bind to all catalytic sites, and in specific conditions, they stimulate a dysregulated degradation of peptides and globular proteins, instead of inhibiting the enzymatic activity, as expected for serine proteases in general. Aiming to explore and explain this paradoxical effect, we solved and refined the crystal structure of native ClpP from Staphylococcus epidermidis (Se), an opportunistic pathogen involved in nosocomial infections, as well as ClpP in complex with ixazomib at 1.90 Å and 2.33 Å resolution, respectively. The interpretation of the crystal structures, in combination with complementary biochemical and biophysical data, shed light on how ixazomib affects the ClpP conformational state and activity. Moreover, SEC-SAXS and DLS measurements show, for the first time, that a peptidomimetic boronate compound also induces the assembly of the tetradecameric structure from isolated homomeric heptameric rings of a gram-positive organism.

Real-World Study of Adjuvant Biosimilar Trastuzumab-dkst for HER2-Positive Breast Cancer Treatment in a Brazilian Population.

INTRODUCTION: Biological monoclonal antibodies play a pivotal role in cancer treatment, with biosimilars significantly enhancing their accessibility. In Brazil's ethnically diverse setting, real-world evidence is crucial for assessing the effectiveness and applicability of these therapies in routine clinical practice. METHODS: We performed a multicentric, observational, prospective real-world study on biosimilar trastuzumab-dkst for adjuvant treatment of early HER2-positive breast cancer in Brazilian patients. Data were collected using a case-report form. RESULTS: Of the 176 recruited, we present data from the first 59 patients (mean age 51.7 ± 12.9 years) who had completed treatment with trastuzumab-dkst. The mean time from diagnosis to the first adjuvant treatment with trastuzumab-dkst was 5.5 ± 2.7 months. Of the patients, 59% of patients achieved at least a 30-month follow-up. The 31.7-month invasive disease-free survival rate (IDFS) was 94.5% (95% CI 83.9-98.2%) and median IDFS was not achieved, since only three patients had invasive disease recurrence. The overall survival rate was 100% until the last assessment. The observed adverse events were similar to those presented by other studies using biosimilar or reference trastuzumab. Four serious adverse events (8.5%) were observed. A reduction in left ventricular ejection fraction of at least 10% was observed in 16.9% of participants. There was no treatment interruption, and three participants (5.1%) had their trastuzumab-dkst dose reduced. CONCLUSION: Our study reinforces the existing pivotal data, underscoring the real-world efficacy and safety of biosimilar trastuzumab-dkst in the adjuvant treatment for early HER2-positive breast cancer. The preliminary long-term effectiveness and safety data we present further validate trastuzumab-dkst's role as a cost-saving alternative in oncological care. These findings have important implications for improving patient access to crucial treatments and for the more efficient use of healthcare resources. GOV REGISTRATION: NCT03892655.

Protease-activated receptor type 2 activation downregulates osteogenesis in periodontal ligament stem cells

Abstract Protease-activated receptor-2 (PAR2) is associated with the pathogenesis of many chronic diseases with inflammatory characteristics, including periodontitis. This study aimed to evaluate how the activation of PAR2 can affect the osteogenic activity of human periodontal ligament stem cells (PDLSCs) in vitro. PDLSCs collected from three subjects were treated in osteogenic medium for 2, 7, 14, and 21 days with trypsin (0.1 U/mL), PAR2 specific agonist peptide (SLIGRL-NH2) (100 nM), and PAR2 antagonist peptide (FSLLRY-NH2) (100 nM). Gene (RT-qPCR) expression and protein expression (ELISA) of osteogenic factors, bone metabolism, and inflammatory cytokines, cell proliferation, alkaline phosphatase (ALP) activity, alizarin red S staining, and supernatant concentration were assessed. Statistical analysis of the results with a significance level of 5% was performed. Activation of PAR2 led to decreases in cell proliferation and calcium deposition (p < 0.05), calcium concentration (p < 0.05), and ALP activity (p < 0.05). Additionally, PAR2 activation increased gene and protein expression of receptor activator of nuclear factor kappa-Β ligand (RANKL) (p < 0.05) and significantly decreased the gene and protein expression of osteoprotegerin (p <0. 05). Considering the findings, the present study demonstrated PAR2 activation was able to decrease cell proliferation, decreased osteogenic activity of PDLSCs, and upregulated conditions for bone resorption. PAR2 may be considered a promising target in periodontal regenerative procedures.

Protease-activated receptor type 1 (PAR1) increases CEMP1 gene expression through MAPK/ERK pathway

Abstract: PAR1 is a G-coupled protein receptor that regulates several cellular metabolism processes, including differentiation and proliferation of osteogenic and cementogenic related cells and our group previously demonstrated the regenerative potential of PAR1 in human periodontal ligament stem cells (hPDLSCs). In this study, we hypothesized that PAR1 regulates the cementogenic differentiation of hPDLSCs. Our goal was to identify the intracellular signaling pathway underlying PAR1 activation in hPDSLC differentiation. hPDLSCs were isolated using the explant technique. Cells were cultured in an osteogenic medium (OST) (α-MEM, 15% fetal bovine serum, L-glutamine, penicillin, streptomycin, amphotericin B, dexamethasone, and beta-glycerophosphate). The hPDLSCs were treated with a specific activator of PAR1 (PAR1 agonist) and blockers of the MAPK/ERK and PI3K pathways for 2 and 7 days. The gene expression of CEMP1 was assessed by RT-qPCR. The activation of PAR1 by its agonist peptide led to an increase in CEMP1 gene expression when compared with OST control. MAPK/ERK blockage abrogated the upregulation of CEMP1 gene expression induced by PAR1 agonist (p < 0.05). PI3K blockage did not affect the gene expression of CEMP1 at any experimental time (p > 0.05). We concluded that CEMP1 gene expression increased by PAR1 activation is MAPK/ERK-dependent and PI3K independent, suggesting that PAR1 may regulate cementogenetic differentiation of hPDLSCs.

Effects of periodontal treatment on primary sjogren's syndrome symptoms

Abstract The aim of this longitudinal prospective study was to evaluate the effects of periodontal treatment on the clinical, microbiological and immunological periodontal parameters, and on the systemic activity (ESSDAI) and subjective (ESSPRI) indexes in patients with primary Sjögren’s Syndrome (pSS). Twenty-eight female patients were divided into four groups: pSS patients with or without chronic periodontitis (SCP, SC, respectively), and systemically healthy patients with or without chronic periodontitis (CP, C, respectively). Periodontal clinical examination and immunological and microbiological sample collection were performed at baseline, 30 and 90 days after nonsurgical periodontal treatment (NSPT). Levels of interleukin IL-1β, IL-8 and IL-10 in saliva and gingival crevicular fluid (GCF) were evaluated by ELISA, as well as the expression of Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans, (Aa) Tannerella forsythia (Tf), and Treponema denticola (Td), by qPCR. Systemic activity and pSS symptoms were evaluated by ESSDAI and ESSPRI. NSPT resulted in improved periodontal clinical parameters in both SCP and CP groups (p>0.05). Pg, Aa, and Tf levels decreased after NSPT only in CP patients (p<0.05). Significantly greater levels of IL-10 in GCF were verified in both SCP and CP groups (p<0.05). SCP patients showed increased salivary flow rates and decreased ESSPRI scores after NSPT. In conclusion, NSPT in pSS patients resulted in improved clinical and immunological parameters, with no significant effects on microbiological status. pSS patients also showed increased salivary flow and lower ESSPRI scores after therapy. Therefore, it can be suggested that NSPT may improve the quality of life of pSS patients.

Efeito da ativação do receptor ativado por protease do tipo 2 (PAR-2) sobre a atividade osteogênica de células mesenquimais do ligamento periodontal / Effect of type 2 protease activated receptor (PAR-2) activation in the osteogenic activity of the periodontal ligament stem cells

O receptor ativado por protease do tipo 2 (PAR-2) está associado à patogênese de doenças inflamatórias crônicas, incluindo periodontite, e a ativação do PAR-2 desempenha papel relevante no processo inflamatório. O objetivo do presente estudo foi avaliar o efeito da ativação do PAR-2 na atividade osteogênica de células-tronco do ligamento periodontal humano (PDLSCs). PDLSCs obtidos de 3 sujeitos foram cultivados em meio controle ou em meio osteogênico por 2, 7, 14 e 21 dias. Proliferação celular, atividade da fosfatase alcalina (ALP), expressão gênica (qPCR) e expressão proteica (ensaio ELISA) de fatores osteogênicos e depósitos de cálcio, concentração de cálcio (sobrenadante) foram avaliados na presença de tripsina (0,1 U/ml), peptídeo agonista de PAR-2 (100nM), peptídeo antagonista de PAR-2 (100 nM) e peptídeo controle de PAR-2. A ativação do PAR-2 levou à alteração na proliferação celular, diminuição na formação de depósitos de cálcio (p <0,05), na concentração de cálcio (p<0,05) e atividade da ALP (p <0,05). Além disso, os ensaios de qPCR e ELISA mostraram que a ativação de PAR-2 pode aumentar a expressão gênica e protéica de RANKL (p<0,05) e diminuir a expressão gênica e proteica de OPG (p<0,05). Os resultados do presente estudo demonstram que a ativação do PAR-2 aumenta a proliferação e diminuem a atividade osteogênica dos PDLSCs, indicando que o PAR-2 pode ser um alvo importante a ser considerado no uso de células mesenquimais do ligamento periodontal em procedimentos regenerativos em periodontia.

Qualidade higiênico sanitária de alfaces (Lactuca sativa) comercializadas em feiras livres na cidade de Uberlândia, MG, Brasil / Hygienic-sanitary quality of the lettuce (Lactuca sativa) commercialized in free fairs in Uberlandia city, MG, Brazil

No Brasil, as parasitoses intestinais estão entre os problemas mais comumente diagnosticados em saúde pública. Essas doenças são amplamente difundidas em virtude da precariedade das condições sanitárias, e as hortaliças servem como um dos principais veículos de sua transmissão. A contaminação dos vegetais pode ocorrer no campo, no período de produção, durante o processo de colheita ou até mesmo na saída do produto em direção à mesa do consumidor, pois os microrganismos podem se multiplicar nos alimentos de acordo com a umidade e temperatura. O objetivo deste trabalho foi avaliar a qualidade de alfaces provenientes de feiras livres do município de Uberlândia, em relação à contaminação microbiológica e parasitológica. De junho a setembro/2009 foram coletadas aleatoriamente 96 amostras de alfaces, da variedade crespa, em 12 bancas de diferentes produtores. Tais amostras foram submetidas ao método de sedimentação espontânea por 24 horas para análise parasitológica. Os resultados obtidos indicaram 100% de contaminação parasitária nas amostras avaliadas, destacando-se Balantidium coli, Entamoeba histolytica/dispar, Giardia lamblia e Cryptosporidium spp. Considerando a elevada frequência de contaminação parasitária e o potencial risco de doenças veiculadas pelas hortaliças, sugere-se uma vigilância sanitária mais atuante na fiscalização de alimentos oferecidos à população, como também um programa de educação sanitária para os produtores e consumidores.

In Brazil the intestinal parasitism is one of the problems normally diagnosed in public health. These diseases are widely spread out in virtue of the precariousness of the sanitary conditions; the vegetables serve as one of the main vehicles of its transmission. The contamination of vegetables can occur in the field, in the period of production, during the process of harvest or even in the exit of the product towards the table of the consumer, because the microorganisms can be multiplied in foods according to the humidity and temperature. The aim of this work was to evaluate the quality of lettuces proceeding from free fairs of Uberlândia city, MG ­ Brazil - in relation to the microbiological and parasitological contamination. From June to September/2009, 96 samples of lettuces had been collected randomly in 12 stalls of different producers. Such samples had been submitted to the method of spontaneous sedimentation for 24 hours for parasitological analysis. The found results indicated 100% of parasitic contamination in the evaluated samples; being distinguished Balantidium coli, Entamoeba histolytica/dispar, Giardia lamblia, and Cryptosporidium spp. Considering the raised frequency of parasitic contamination and the potential risk of illnesses propagated by the vegetables, a more operating sanitary monitoring in the fiscalization of foods offered to the population is suggested, as well as a program of sanitary education for the producers and consumers.

Uso do Anel de Reforço Muller: Para Falha Acetabular na Revisão de Artroplastia Total de Quadril

O anel de reforço acetabular de Muller (ARM)é mais um componente no arsenal das artroplastias totais de quadril,sejam, primárias ou revisões. Este dispositivo, é indicado nas displasias acetabulares primárias e revisões acetabulares associadas à defeitos segmentares periféricos