Type 1 diabetes mellitus (T1DM) represents one of the most frequent chronic illnesses affecting children. The early diagnosis of this disease is crucial, as it plays a key role in preventing the development of a life-threatening acute complication: diabetic ketoacidosis. The etiopathogenetic role of viral infections has long been suggested and emerging data are pointing towards a complex bidirectional relationship between diabetes and COVID-19. The aim of this study is to assess the impact of the COVID-19 pandemic on the incidence and severity of new T1DM cases in children in Romania. We analyzed the differences between a group of 312 patients diagnosed with T1DM in the period 2003–2019 and a group of 147 children diagnosed during the pandemic. The data were investigated using statistical analysis of a series of relevant variables. The total number of newly diagnosed T1DM increased by 30.08% in the period March 2020–February 2021 compared to the previous years. The patients in the pandemic group had a higher mean age at the onset of T1DM, were less frequently living in an urban area, and presented a higher mean value of HbA1c. Diabetic ketoacidosis at the onset of T1DM was 67.40% more frequent, and a higher percentage of these patients presented with a severe form. The duration of T1DM symptoms did not differ significantly between the two groups. A number of 8 patients associated SARS-CoV-2 infection at the time of T1DM diagnosis.

Human leukemia Jurkat T cells were analyzed for apoptosis and cell cycle by flow cytometry, using the Annexin V/propidium iodide (PI) standard assay, and a simple PI staining in Triton X-100/digitonin-enriched PI/RNase buffer, respectively. Cells treated with doxorubicin or menadione displayed a very strong correlation between the apoptotic cell fraction measured by the Annexin V/PI assay, and the weight of a secondary cell population that emerged on the forward scatter (FS)/PI plot, as well as on the side scatter (SS)/PI and FL1/PI plots generated from parallel cell cycle recordings. In both cases, the Pearson correlation coefficients were >0.99. In cell cycle determinations, PI fluorescence was detected on FL3 (620/30 nm), and control samples exhibited the expected linear dependence of FL3 on FL1 (525/40 nm) signals. However, increasing doses of doxorubicin or menadione generated a growing subpopulation of cells displaying a definite right-shift on the FS/FL3, SS/FL3 and FL1/FL3 plots, as well as decreased PI fluorescence, indicative of ongoing fragmentation and loss of nuclear DNA. By gating on these events, the resulting fraction of presumably sub-cycling cells (i.e. cells with cleaved DNA, counting sub-G₀/G₁, sub-S and sub-G₂/M cells altogether) was closely similar to the apoptotic rate assessed by Annexin V/PI labeling. Taken together, these findings suggest a possible way to recognize the entire population of cells undergoing apoptotic DNA cleavage and simultaneously determine the cell cycle distribution of non-apoptotic cells in PI-labeled cell samples with various degrees of DNA fragmentation, using a simple and reproducible multiparametric analysis of flow cytometric recordings.

One of the most widespread bacterial infections worldwide, Helicobacter pylori is thought to affect almost half of the world’s population. Due to rising antibiotic resistance, treatment should be tailored according to antibiotic susceptibility testing (AST). This study aims to evaluate Helicobacter pylori antibiotic resistance and its therapeutic efficacy in children. We conducted a prospective, single-center study, that evaluated 68 children referred for upper gastrointestinal endoscopy (UGE) following chronic dyspeptic syndrome. Each patient underwent multiple biopsies to perform bacterial cultures with AST and histopathological examinations for the diagnosis. Patients without antibiotic resistance or negative cultures received a 10-day sequential therapy, while the others had the antibiotic regimen tailored based on AST. Fifty-nine patients with a positive biopsy-based diagnosis (24 males) were finally included. Bacterial cultures with AST were positive for 13 patients (22.03%) and the antibiotic resistance for clarithromycin was 15.38%. Fifty-seven patients were administered sequential therapy with an eradication rate of 94.73%. Clarithromycin-resistant patients were successfully treated with 10-day triple therapy of esomeprazole, amoxicillin, and metronidazole. Although bacterial cultures had a low positivity rate, sequential therapy had a successful eradication rate. Further studies are necessary to better assess Helicobacter pylori antibiotic resistance to provide tailored treatment and identify children that need closer monitoring.

Children with inflammatory bowel disease (IBD) have an increased susceptibility to Clostridium difficile infection (CDI), with a rising incidence over time. Differentiating between CDI and IBD exacerbation is challenging due to overlapping symptoms. In our cohort of 55 pediatric IBD patients, 6 were diagnosed with CDI. Upon conducting a thorough patient evaluation and subsequent data analysis, an exhaustive review of the existing literature was undertaken. CDI is more prevalent in ulcerative colitis (UC) than Crohn’s disease (CD) patients, as seen in our patients and in the existing literature. The management of a pediatric patient with IBD is itself a challenge for a clinician because of the chronic, possibly relapsing course, and substantial long-term morbidity. When CDI is added, it becomes even more demanding, since CDI leads to more severe disease in children with IBD. A multidisciplinary approach and intensive treatment for possible sepsis, anemia, hypoalbuminemia, and hydro-electrolytic and acid-base imbalances are frequently mandatory in patients with CDI and IBD, which leads to a significant health care burden in hospitalized children with IBD. After the infection is treated with antibiotic therapy, important considerations regarding the future treatment for the underlying IBD are also necessary; in most cases, a treatment escalation is required, as also seen in our study group.

Food allergies represent immune-mediated adverse reactions to dietary proteins, with incompletely elucidated causes and etiopathogenic mechanisms, and have a significant negative impact on the quality of life of children and their families. The aim of the study was to assess the associations between IL-13 gene single-nucleotide polymorphisms (SNPs) rs20541 and rs1800925 and the risk of developing IgE-mediated or non-IgE-mediated food allergies, as well as other patient characteristics. The study included 115 healthy controls and 165 children with food allergy, divided into two groups based on the presence of either IgE-mediated (N=85) or non-IgE-mediated (N=80) food allergy, all of Romanian origin. Genotyping of the two SNPs was performed using Real-Time PCR with TaqMan Allelic Discrimination Assays. OpenEpi and PLINK v1.07 were used for the statistical analysis. The frequency of carriers of the minor allele T of rs1800925 was significantly higher in patients with non-IgE-mediated food allergy (50% vs. 35.6%, p=0.04, OR=1.8). CT heterozygotes of rs1800925 were at high risk for non-IgE-mediated food allergy compared to CC and TT homozygotes (p=0.01, OR=2.13). No associations were found with IgE-mediated food allergy. The present study investigates for the first time IL-13 gene SNPs in non-IgE-mediated food allergy. The results provide evidence that variation in the IL-13 gene is involved in the pathogenesis of this disease. Further investigation is required to confirm these findings.