Molecular diagnostic of complicated pneumonia in the post-vaccine era.

BACKGROUND: The etiological diagnosis of community-acquired pneumonia (CAP) is still a challenge. We compared the conventional culture method and real-time polymerase chain reaction (RT-PCR) for the identification of Streptococcus pneumoniae in severe pediatric CAP. METHODS: A retrospective hospital-based study was conducted. From 2012 to 2018, we have selected patients who had peripheral blood and/or pleural fluid collected for etiological investigation by RT-PCR. RESULTS: We included 113 children (median age: 3 years; interquartile range 1-6 years). RT-PCR increased the detection rate of S. pneumoniae by 6.5 times using blood samples and eight times using pleural fluid samples. Patients subjected to RT-PCR showed more prolonged hospitalization (p = 0.006), fewer comorbidities (p = 0.03), presence of pleural effusion (p = 0.001), presence of young forms of leukocytes (p = 0.001) and radiograph with characteristics of pneumonia (p = 0.002). The presence of pleural effusion [odds ratio (OR) = 14.7, 95% confidence interval (CI) 1.6-133.9; p = 0.01] and young forms of leukocytes (OR = 8.9, 95% CI 0.9-84.4; p = 0.05) were risk factors for positive RT-PCR pneumococcal when multivariate analysis was performed. CONCLUSIONS: RT-PCR is a reliable method for diagnosing severe CAP using sterile materials and a potentially applicable method in patients with clinical, radiological and non-specific laboratory characteristics of lower respiratory tract infection, especially in complicated cases with pleural effusion.

Complete enucleation surgical treatment and histological analysis of a mucocele in lower lip in a child: case report / Tratamiento quirúrgico de enucleación completa y análisis histológico de un mucocele en el labio inferior en un niño: reporte de caso

Objective: The aim of this case report is to describe the surgical removal of a mucocele and its histological analysis, in a child. Case Report: An 11-year-old female patient attended the Pediatric Dentistry clinic complaining of a lower lip lesion. During the anamnesis, the mother reported that the child had a habit of biting and sucking the spot frequently. Clinical examination showed the lesion was compatible with a mucocele. The proposed treatment was a complete enucleation of the lesion under local anesthesia. The incision and tissue divulsion were performed for maximum preservation of the mucosa, avoiding a possible recurrence. Total adjacent glands removal was also performed. The lesion was placed in 10% formaldehyde for histopathological analysis (H&E Staining), which showed dense connective tissue presenting chronic inflammatory infiltrate and extravasated mucin, presence of granulation tissue delimiting the area of extravasated mucin and presence of minor salivary glands. The patient was advised to quit the habit, and after seven days the sutures were removed. At the one-year follow-up there was no recurrence of the lesion. Conclusion: The proposed treatment proved to be effective without recurrence of the lesion.

Objetivo: El objetivo de este reporte de caso es describir la extirpación quirúrgica de un mucocele y su análisis histológico en un niño. Informe del caso: una paciente de 11 años de edad asistió a la clínica de Odontopediatria quejándose de una lesión en el labio inferior. Durante la anamnesis, la madre informó que el niño tenía la costumbre de morder y chupar el lugar con frecuencia. En el examen clínico, la lesión fue compatible con un mucocele. El tratamiento propuesto fue una enucleación completa de la lesión bajo anestesia local. La incisión y la divulgación del tejido se realizaron para la máxima preservación de la mucosa, evitando una posible recurrencia. También se realizó la extracción total de las glándulas adyacentes. La lesión se colocó en formaldehído al 10% para el análisis histopatológico (tinción H&E), que mostró tejido conectivo denso que presenta infiltrado inflamatorio crónico y mucina extravasada, presencia de tejido de granulación que delimita el área de mucina extravasada y presencia de glándulas salivales menores. Se aconsejó al paciente que abandonara el hábito, y después de siete días se retiraron las suturas. En el seguimiento de un año no hubo recurrencia de la lesión. Conclusión: El tratamiento propuesto demostró ser efectivo sin recurrencia de la lesión.

Resveratrol Regulates the Quiescence-Like Induction of Activated Stellate Cells by Modulating the PPARγ/SIRT1 Ratio.

The activation of hepatic stellate cell (HSC), from a quiescent cell featuring cytoplasmic lipid droplets to a proliferative myofibroblast, plays an important role in liver fibrosis development. The GRX line is an activated HSC model that can be induced by all-trans-retinol to accumulate lipid droplets. Resveratrol is known for activating Sirtuin1 (SIRT1), a NAD(+)-dependent deacetylase that suppresses the activity of peroxisome proliferator-activated receptor gamma (PPARγ), an important adipogenic transcription factor involved in the quiescence maintenance of HSC. We evaluated the effects of 0.1 µM of resveratrol in retinol-induced GRX quiescence by investigating the interference of SIRT1 and PPARγ on cell lipogenesis. GRX lipid accumulation was evaluated through Oil-red O staining, triacylglycerides quantification, and [(14)C] acetate incorporation into lipids. mRNA expression and protein content of SIRT1 and PPARγ were measured by RT-PCR and immunoblotting, respectively. Resveratrol-mediated SIRT1 stimuli did not induce lipogenesis and reduced the retinol-mediated fat-storing capacity in GRX. In order to support our results, we established a cell culture model of transgenic super expression of PPARγ in GRX cells (GRXPγ). Resveratrol reduced lipid droplets accumulation in GRXPγ cells. These results suggest that the PPARγ/SIRT1 ratio plays an important role in the fate of HSC. Thus, whenever the PPARγ activity is greater than SIRT1 activity the lipogenesis is enabled.

The interplay between apoptosis, mitophagy and mitochondrial biogenesis induced by resveratrol can determine activated hepatic stellate cells death or survival.

Resveratrol has been the focus of numerous studies reporting opposite effects that depend on its concentration. The GRX is an activated hepatic stellate cells model used to study liver fibrosis development and resolution. We recently showed that GRX treatment with RSV (0.1-50 µM) for 24 h triggered dose-dependent pro-oxidant effects, resulting in cytotoxicity and cell damage only at the highest concentration. Here, we evaluated whether the pro-oxidant effect of resveratrol treatment is accompanied by alterations on the GRX mitochondrial metabolism, and whether the concomitantly autophagy/mitophagy induction can influence on cell death or survival. We demonstrated that all concentrations of resveratrol promoted an increase of GRX cell death signals, altering the mitochondrial dynamics and function. Cells treated with all resveratrol concentrations presented higher autophagy/mitophagy features, but only treatments with 1 and 10 µM of resveratrol-induced mitochondrial biogenesis. Since cell damage was higher and there was no mitochondrial biogenesis in GRX treated with 50 µM of resveratrol, we suggest that these cells failed to remove and replace all damaged mitochondria. In conclusion, the cytotoxic effect of resveratrol that effectively promotes cell death could be related to the interrelation between the concomitant induction of apoptosis, autophagy/mitophagy and mitochondrial biogenesis in GRX.