Exhausted and Apoptotic BALF T Cells in Proinflammatory Airway Milieu at Acute Phase of Severe Mycoplasma Pneumoniae Pneumonia in Children

Severe mycoplasma pneumoniae pneumonia (MPP) in children presents with serious clinical complications. Without proper and prompt intervention, it could lead to deadly consequences. Dynamics of the inflammatory airway milieu and activation status of immune cells were believed to be the hallmark of the pathogenesis and progress of the disease. In this study, by employing the T-cell sorting and mRNA microarray, we were able to define the main feature of the chemokine/cytokine expression and the unique characteristics of T cells in the bronchoalveolar lavage fluid (BALF) from severe MPP patients at acute phase. Our study for the first time delineated the molecular changes in isolated BALF T cells in severe MPP children with respect to the cytokine/chemokine expression, cell activation, exhaustion, and apoptosis. By comparing the BALF aqueous expression of cytokines/chemokines with that in sorted T cells, our data give a preliminary clue capable of finishing out the possible cell source of the proinflammatory cytokines/chemokines from the BALF mixture. Meanwhile, our data provide a distinctively pellucid expression profile particularly belonging to the isolated BALF T cells demonstrating that in the inflammatory airway, overactivated T cells were exhausted and on the verge of apoptotic progress.

After A Year of Follow Up Non-Cirrhotic Portal Hypertension Patient with Partial Spleen Embolization (PSE) Management

Non-cirrhotic portal hypertension (NCPH) is a heterogeneous group of liver disorders leading to portal hypertension. There are multiple approaches to managing portal hypertension' clinical complications to treat/prevent spontaneous hemorrhage by mitigating thrombocytopenia. Portal hypertension complications have been traditionally managed with serial endoscopic variceal ligation (EVL) or with invasive open surgical procedures such as orthotopic liver transplantation (OLT) or portosystemic shunting, splenectomy.6–9 There are several risks associated with splenectomies, such as hemorrhagic complications or intraoperative blood loss.5,6,14 Partial Spleen Embolization (PSE) ‎may overcome the limitations of splenectomy and provide patients with an alternative treatment. An eighteen-year-old male has a splenomegaly history since he was 12 years old and has recurring hematemesis and melena. After performing abdominal computed tomography, laboratory studies, and several endoscopies, the results indicated secondary hypersplenism due to non-cirrhotic portal hypertension. The patient had 13 endoscopies and 2 EVL in 5 years. Despite adequate treatment, the patients developed recurrent variceal bleeding and no improvement in blood function. The patient underwent PSE at Integrated Cardiovascular Center in Dr. Cipto Mangunkusumo, General Hospital, Jakarta, Indonesia. It was performed through the femoral access with a PVA (polyvinyl alcohol) embolus. The procedure went successful, and there was no major complication with the patient. Twenty days after the patient had an abdominal CT scan, it showed no abscess, and the spleen volume was reduced by 20%. Long-term results over a year after the procedure are presented. PSE is a safe, effective, semi-invasive alternative to splenectomy in non-cirrhotic portal hypertension because it preserves functional spleen mass and avoids postprocedure accelerated liver disease or encephalopathy.

Bone Union of Osseous Microvascular Free Tissue Transfer in Mandibular Reconstruction

Objectives Osseous microvascular free tissue transfer (MFTT) is the gold standard for reconstruction for most segmental mandibulectomy defects. The most common osseous MFTT utilized in reconstruction is the fibular, scapular, and osteocutaneous radial forearm (OCRF) free flap. We evaluated postoperative bone union as well as clinical complications following MFTT and the impact of various patient and reconstructive characteristics, including type of osseous MFTT. Study Design Retrospective cohort study. Setting Tertiary care academic hospital. Methods This study examined patients who underwent osseous MFTT for mandibular defects from January 2017 to January 2019. Results An overall 144 osteotomies in 58 patients were evaluated. Of the 144 junctions, 28 (19.4%) showed radiographic nonunion. Patients who underwent preoperative (odds ratio [OR] = 0.30, P = .027) and postoperative (OR = 0.28, P = .003) radiation had a significantly lower bone union score. Time from surgery to postoperative imaging was associated with higher bone union scores (OR = 1.07, P = .024). When bone union scores were compared among types of MFTT, fibular (OR = 5.62, P = .008) and scapular (OR = 4.69, P = .043) MFTT had significantly higher scores than OCRF MFTT. Twelve (20.7%) patients had postoperative complications. There was no statistically significant correlation between clinical complications and various variables, including type of osseous MFTT. Conclusion Pre- and postoperative radiation and time from surgery have an impact on bone union. Regarding the type of MFTT, fibular and scapular MFTT appeared to have higher bone union when compared with OCRF. There was no impact of bone union or type of osseous MFTT on clinical complications.

Use of recombinant human deoxyribonuclease in pediatric intensive care unit – a single-center experience

ABSTRACT Objective: Dornase alfa (rhDNase) reduces the viscosity of purulent sputum in the lungs. The use in patients with cystic fibrosis (CF) is proven. However, the evidence of its applicability to other conditions is limited. This study aims to present the authors’ experience with the use of rhDNase in non-CF patients admitted to the Pediatric Intensive Care Unit (PICU). At the study center, rhDNase was used during flexible bronchoscopies in 24 cases, of which 20 (83%) had atelectasis and seven (29%) were admitted to PICU. Four patients (57%) were on invasive mechanical ventilation (MV). Case description: Two cases of daily rhDNase administration at PICU are presented: patient A was an 8-year-old boy admitted with septic shock and acute respiratory distress syndrome (ARDS). The patient required mechanical ventilation with aggressive settings and experienced several clinical complications. On D50, he started rhDNase treatment with an improvement in FiO2, PaCO2 and PaO2/FiO2 ratio according to radiologic findings. He was extubated on D23 of treatment. Patient B was a 17-month-old girl admitted with a convulsive status epilepticus who experienced respiratory complications (infectious and barotrauma) with ARDS, requiring aggressive ventilation. She initiated rhDNase treatment on D60. During the treatment an improvement in FiO2, PaO2/FiO2 ratio and a tendency of PaCO2 decrease were found. She had radiological improvement. No complications were described. Comments: RhDNase may be a helpful and safe tool to use in PICU prolonged intubated patients with ventilator-induced lung injury. Further studies are needed to assess and propose valid indications.

CRISPR/CasRx Proof-of-Concept for RNA Degradation: A Future Tool against RNA Viruses?

Influenza viruses provide a great threat for the human population, causing highly contagious respiratory infections that can lead to serious clinical complications. There are a limited variety of influenza antivirals, and these antivirals are subjected to the constant emergence of resistances. Therefore, the development of new antiviral strategies to combat influenza viruses and other RNA viruses must be promoted. In this work, we design a proof-of-concept of a recently described CRISPR/Cas tool that has been proposed as a possible future RNA virus antiviral, named CRISPR/CasRx. For this, we verified the efficiency of the CasRx endonuclease in the degradation of the eGFP mRNA reporter gene and we established the best conditions for, and the efficient performance of, the CRISPR/CasRx system. The results were measured by fluorescence microscopy, flow cytometry, and qRT-PCR. The analyses demonstrated a reduction in fluorescence, regardless of the amount of eGFP reporter plasmid transfected. The analyses showed an 86–90% reduction in fluorescence by flow cytometry and a 51–80% reduction in mRNA expression by qRT-PCR. Our results demonstrate that the CasRx endonuclease is an efficient tool for eGFP mRNA knockdown. Therefore, subsequent experiments could be useful for the development of a new antiviral tool.

Pragmatic approach to rehabilitate maxillary teeth in aesthetic zone using resin bonded FDP- A case report

Resin bonded fixed dental prosthesis offers most conservative and cost-effective option with missing teeth in aesthetic zone specially when an implant prosthesis is not indicated. This case report describes a treatment option for the replacement of a missing maxillary central incisor using a double-retainer resin-bonded fixed partial denture (RBFPD), fabricated from zirconium dioxide (ZrO) ceramic. No clinical complications were observed at follow-up examination after placement of the prosthesis. Satisfactory functional and aesthetic results were achieved. A treatment modality using a ZrO ceramic RBFPD is an alternative for single anterior tooth replacement.

Innate and Adaptive Immunity Imbalance With Severe COVID-19 Pneumonia in Children and Adults

Introduction: Little is known about the laboratory and radiological characteristics and clinical significance of peripheral immune alterations in patients with coronavirus disease 2019 (COVID-19). This study aims to clarify these aspects in children and adults with COVID-19.Methods: In this consecutive pilot study, COVID-19 patients with the confirmed pneumonia and real-time RT-PCR were recruited prospectively in June 2020. The clinical, chest CT, and laboratory features, such as lymphocyte subpopulations, were analyzed for each individual.Results: Forty confirmed COVID-19 patients, 11 severe children, 12 severe adults, and 17 critical adult patients, besides 20 healthy pediatrics and 14 healthy adults as controls, were enrolled prospectively. Adult patients, especially critical ones, had a much higher prevalence of laboratory and chest CT abnormalities. Data regarding immune cell subsets in children patients, compared with matched controls, had higher CD3+ CD8+ T cells (p = 0.004) and lower CD4+/CD8+ ratio (p = 0.042), while adult patients, compared with matched controls, had lower CD14+ monocytes (p = 0.032). Adult patients were also categorized as experiencing critical or severe illness on admission and, compared with severe patients, had lower total lymphocytes (p < 0.047), CD3+ T-lymphocytes (p < 0.002), and CD3+ CD8+ T cells (p = 0.001) and, on the other hand, had higher CD3+ CD4+ T cells (p = 0.012) and CD4+/CD8+ ratio (p = 0.003). Non survived adults, compared with survived patients, had significantly lower CD3+ T-lymphocyte (p = 0.005).Conclusion: Unlike adult patients, who compared with matched controls and had more comorbidities, higher frequency of severe clinical symptoms, laboratory abnormalities, and immune cells alteration, clinical manifestations of COVID-19 in children (compared with matched controls) were relatively mild, and fewer clinical complications were seen either, perhaps because of a milder inflammatory response following their peripheral innate and adaptive immune cell alteration pattern.

Root and Root Canal Morphology: Study Methods and Classifications

Background: Morphology of the root canal system is divergent and unpredictable, and rather linked to clinical complications, which directly affect the treatment outcome. This objective necessitates continuous informative update of the effective clinical and laboratory methods for identifying this anatomy, and classification systems suitable for communication and interpretation in different situations. Data: Only electronic published papers were searched within this review. Sources: “PubMed” website was the only source used to search for data by using the following keywords "root", "canal", "morphology", "classification". Study selection: 153 most relevant papers to the topic were selected, especially the original articles and review papers, from 1970 till the 28th of July 2021. Conclusions: This review divided the root canal analysis methods into two approaches; clinical and in vitro techniques. The latter has shown more precise non-subjective readings, on the other hand; the clinical methods provide direct chair side diagnosis for the clinical cases. The classification systems reviewed in the present study, started with the oldest trials that simply presented the root canal systems, according to the degree of angulation, or by coded Latin numbers or English letters. Then, the most recent systems were also presented that were persisted with continuous editions up to date. These new systems could briefly describe the root and root canal’s internal and external details in a small formulation, without confusion and in an easily communicated manner, highly recommended specially for students, teachers, and researchers

New Mutations in HFE2 and TFR2 Genes Causing Non HFE-Related Hereditary Hemochromatosis

Hereditary hemochromatosis (HH) is an iron metabolism disease clinically characterized by excessive iron deposition in parenchymal organs such as liver, heart, pancreas, and joints. It is caused by mutations in at least five different genes. HFE hemochromatosis is the most common type of hemochromatosis, while non-HFE related hemochromatosis are rare cases. Here, we describe six new patients of non-HFE related HH from five different families. Two families (Family 1 and 2) have novel nonsense mutations in the HFE2 gene have novel nonsense mutations (p.Arg63Ter and Asp36ThrfsTer96). Three families have mutations in the TFR2 gene, one case has one previously unreported mutation (Family A—p.Asp680Tyr) and two cases have known pathogenic mutations (Family B and D—p.Trp781Ter and p.Gln672Ter respectively). Clinical, biochemical, and genetic data are discussed in all these cases. These rare cases of non-HFE related hereditary hemochromatosis highlight the importance of an earlier molecular diagnosis in a specialized center to prevent serious clinical complications.