Chemistry and Biochemistry Aspects of the 4-Hydroxy-2,3-trans-nonenal

4-hydroxy-2,3-trans-nonenal (C9H16O2), also known as 4-hydroxy-2E-nonenal (C9H16O2; HNE) is an α,β-unsaturated hydroxyalkenal. HNE is a major aldehyde, formed in the peroxidation process of ω-6 polyunsaturated fatty acids (ω-6 PUFAs), such as linoleic and arachidonic acid. HNE is not only harmful but also beneficial. In the 1980s, the HNE was regarded as a “toxic product of lipid peroxidation” and the “second toxic messenger of free radicals”. However, already at the beginning of the 21st century, HNE was perceived as a reliable marker of oxidative stress, growth modulating factor and signaling molecule. Many literature data also indicate that an elevated level of HNE in blood plasma and cells of the animal and human body is observed in the course of many diseases, including cancer. On the other hand, it is currently proven that cancer cells divert to apoptosis if they are exposed to supraphysiological levels of HNE in the cancer microenvironment. In this review, we briefly summarize the current knowledge about the biological properties of HNE.

VEGF and IL-6 correlation in POEMS: a potential upcoming marker of active disease and early autologous BMT response.

Introduction: VEGF function may be responsible for most POEMS manifestations, and it is considered a reliable marker of disease. COVID-19 era arose increasing interest for other inflammatory cytokines, with particular focus on Interleukin-6; VEGF production is stimulated by IL-6 and IL1β, whose concentrations appear to be elevated in clonal plasma cells. Objectives: This study aims to simultaneously evaluate VEGF and IL-6 values in patients (pts) with POEMS at different stages of the disease to find a correlation between them. Methods: We performed a monocentric study, measuring serum levels of VEGF and IL-6 in 8 POEMS pts at different time points of the disease. Results: We observed elevated serum levels of both VEGF and IL-6 in three pts before transplant, while the day after the infusion of autologous stem cells, we observed a steep decrease of both serum markers. Among the four-pts tested only after transplant, two presented with a consensual level of VEGF and IL-6, while the others did not correlate. One patient observed at POEMS diagnosis, during active disease, presented with strikingly high levels of both serum markers. Conclusions: So far, to the best of our knowledge, IL-6 could be considered as a marker of active disease and reliable up to the very first months after BMT, after which its accuracy appears to be lost due to unknown factors, still to be investigated.

Myelin Protein Zero Immunohistochemistry Is Not a Reliable Marker of Extrinsic Mucosal Innervation in Patients With Hirschsprung Disease

Background Innervation of aganglionic rectum in Hirschsprung disease derives from extrinsic nerves which project from cell bodies located outside the bowel wall and markers that distinguish extrinsic from intrinsic innervation are diagnostically useful. Myelin protein zero (MPZ) is a putative marker of extrinsic glial cells which could distinguish mucosal innervation in aganglionic vs ganglionic colon. Methods Sections and protein blots from ganglionic and aganglionic colon were immunolabeled with MPZ-specific antibodies. Results Immunolabeling of MPZ with a chicken polyclonal or mouse monoclonal antibody confirmed glial specificity and reliably labeled hypertrophic submucosal nerves in Hirschsprung disease. In contrast, a rabbit polyclonal antibody strongly labeled extrinsic and intrinsic nerves, including most mucosal branches. Immunoblots showed MPZ is expressed in mucosal glial cells, albeit at lower levels than in extrinsic nerves, and that the rabbit antibody is more sensitive that the other two probes. Unfortunately, none of these antibodies consistently distinguished mucosal innervation in aganglionic vs ganglionic rectum Conclusions The results suggest that (a) glial cell myelin protein zero expression is influenced more by location (mucosa vs submucosa) than the extrinsic vs intrinsic origin of the accompanied nerves and (b) myelin protein zero immunohistochemistry has limited value as a diagnostic adjunct for Hirschsprung disease.

Umbilical Cord Procalcitonin to Detect Early-Onset Sepsis in Newborns: A Promising Biomarker

Background: Up to 7% of neonates born in high-income countries receive antibiotics for suspected early-onset sepsis (EOS). Culture-proven neonatal sepsis has a prevalence of 0.2%, suggesting considerable overtreatment. We studied the diagnostic accuracy of umbilical cord blood and infant blood procalcitonin (PCT) in diagnosing EOS to improve antibiotic stewardship.Methods: Umbilical cord blood PCT was tested in newborns ≥ 32 weeks of gestation. Groups were defined as following: A) culture-proven or probable EOS (n = 25); B) Possible EOS, based on risk factors for which antibiotics were administered for <72 h (n = 49); C) Risk factor(s) for EOS without need for antibiotic treatment (n = 181); D) Healthy controls (n = 74). Additionally, venous or capillary blood PCT and C-reactive protein (CRP) were tested if blood drawing was necessary for standard care.Results: Between June 2019 and March 2021, 329 newborns were included. Umbilical cord blood PCT was significantly higher in group A than in group C and D. No difference between venous or arterial samples was found. Sensitivity and specificity for cord blood procalcitonin were 83 and 62%, respectively (cut-off 0.1 ng/mL). Antepartum maternal antibiotic administration was associated with decreased PCT levels in both cord blood and infant blood directly postpartum in all groups combined.Conclusion: Umbilical cord blood PCT levels are increased in newborns ≥32 weeks with a proven or probable EOS and low in newborns with risk factors for infection, but PCT seems not a reliable marker after maternal antibiotic treatment. PCT could be useful to distinguish infected from healthy newborns with or without EOS risk factors.

PHOX2B as a Reliable Marker for Neuroblastoma in Tissue and Cytology Specimens

To investigate the diagnostic utility of immunohistochemistry for paired-like homeobox 2B (PHOX2B) expression in neuroblastomas (NBs) and tumors that mimic them, tissue samples (n = 229) from 157 cases of NB, 210 central nervous system tumors, and 170 extracranial non-NB solid tumors (n = 170) were immunostained for PHOX2B. Additionally, PHOX2B expression in 67 body fluid cytology specimens was analyzed. In tissue specimens, PHOX2B expression was positive in NBs, pheochromocytomas, and paragangliomas but negative in all of the other tumors evaluated. PHOX2B was detected by immunohistochemistry in 5 NB cytology specimens; all of the others were negative. These results suggest that PHOX2B may be a sensitive and specific immunohistochemical marker for the pathological diagnosis and differential diagnosis of NB in both tissue and cytology specimens.

The Donor – Recipient Weight Ratio is a Reliable Marker for Cell Yield in Hematopoietic Stem Cell Donations

Bone marrow transplants remain an import source of hematopoietic stem cells for patients suffering from specific diseases like aplastic anemia, for pediatric patients with malignant and non-malignant blood cell disorders, and for situations in which graft-versus-host disease (GvHD) is a concern. Identifying the optimal donor to achieve a 3-5 x 108/kg of recipient weight TNC yield may be challenging. In an analysis of 687 consecutive donors, donor and procedure characteristics were related to TNC/kg of recipient weight using Spearman correlation coefficients as well as linear and multiple regression analysis. We found correlations between donor WBC (r = 0.17), donor platelet counts (r = 0.15), donor BMI (r = 0.10), and the percentage of donor estimated blood volume accessed for harvesting (r = -0.57) with TNC/kg of recipient weight. The strongest correlation existed between the donor-recipient weight ratio and the TNC/kg (r = 84). In a multivariate regression analysis, the donor-recipient weight ratio influenced the TNC/kg of recipient weight more significantly (adjusted R2 = 0.84) than all other related variables put together. The minimal donor-recipient weight ratio associated with a TNC/kg of at least 3x108/kg of recipient weight was 0.8 (mean 3.425; 95% CI 2.01, 5.8). Using this donor-recipient ratio provides national bone marrow donor registries with a practical and simple measure to assure optimal cell yield outcomes in hematopoietic stem cell donations.

Clinical and functional features of the small circle of blood circulation in children with a new form of bronchopulmonary dysplasia

The aim of the work is to analyze the data of Echo-CG examination of premature infants who have formed and have not formed bronchopulmonary dysplasia (BPD) to determine the frequency of the formation of pulmonary hypertension (PH). Materials and methods. A total of 199 preterm infants treated in the Department of Pathology of Newborns were examined. The first group included moderate and severe BPD children (n = 117; 59%). The second group consisted of children without BPD within clearly decreed terms (n = 82; 41%). In each group, patients were divided into four subgroups by the gestational age at birth and the timing of the Echo-CG. Results. Only two (1.1%) patients out of 117 BPD children of the first group were diagnosed with pulmonary hypertension (PH). In 3 (2.5%) of 117 infants of the same group, enlargement of the right heart without PH was revealed. Out of 82 children without BPD, two patients had signs of right heart enlargement. PH was not diagnosed in any of the patients in this group. Discussion. To aggravate the efficiency of PH diagnosis, a number of indices of screening echocardiography seem to be increased by additional analysis of changes in the systolic eccentricity index (EI), as a reliable marker of PH in BPD children. Systolic IE should be integrated into screening in preterm infants for the diagnosis of PH. The use of Echo-CG data along with analysis of blood BNP or NT-proBNP blood content be also expanded. Optimizing the diagnosis of PH at the early stages of BPD is necessary to increase the efficacy of targeted therapy and reduce the risk of severe complications of BPD.

Mouth breathing – A predictor for patient satisfaction after nasal septoplasty?

Background: No reliable marker exists to predict septoplasty outcome. Most patients suffering from nasal airway obstruction (NAO) caused by a deviation of the nasal septum report a bothersome mouth breathing and dryness. In this study our aim was to assess, whether mouth breathing could be objectified in these patients and whether mouth breathing could predict septoplasty outcome. Methods: A monocentric, prospective case-control study of 21 patients was conducted. The proportion of mouth breathing was measured in a blinded manner. As a measurement of patient satisfaction, subjective symptoms pre- and postoperatively, were assessed by using VAS, NOSE and SNOT-20 score. In the patient group an additional acoustic rhinometry and a clinical examination of the nose were performed. Results: With a mean of 25% (SD = 20%) the proportion of mouth breathing in patients with NAO did not differ significantly from the proportion in controls without NAO, with a mean of 27% (SD = 23%). Analysis of subjective scores revealed a significant reduction of subjective symptoms after septoplasty. A higher preoperative proportion of mouth breathing correlated with more remaining postoperative NAO. Conclusions: The percentage of mouth breathing is no different in patients with symptomatic septal deviation than in control patients. Mouth breathing in patients with NAO, evaluated for septoplasty, could be a negative predictive factor for patient satisfaction after nasal septoplasty. Mouth breathing in these patients should be observed carefully because more preoperative mouth breathing should make one more hesitant to consider septoplasty.

Identification of Pre-Gmps and Pre-Meps: Two Novel Types of Committed Progenitor Cells in Bone Marrow

Elucidating the stepwise differentiation processes that leads from multipotent hematopoietic stem cells to mature effector cells is critical for understanding both normal and neoplastic hematopoiesis. Early studies suggested that common myeloid progenitors (CMPs) are oligo-lineage hematopoietic progenitors that produce all lineages of myeloid cells, including granulocytes, monocytes, erythrocytes and megakaryocytes. CMPs do so by first giving rise to megakaryocyte-erythroid progenitors (MEPs) and granulocyte-monocyte progenitors (GMPs), two types of bi-lineage progenitors. However, this concept was challenged by several recent studies where single cell techniques demonstrated that CMPs, GMPs and MEPs are highly heterogenic. The existence of lineage-restricted subsets within the CMP population leads to questions about whether erythroid and megakaryocytic lineage commitment is actually initiated at the multipotent progenitor or CMP stage. During the past 15 years, several lineage-restricted subsets of progenitors have been separated out from CMP population, including monocyte-dendritic progenitors, megakaryocyte progenitors, and erythroid progenitors based on expression of CD115/Flt3, CD41/CD150, and CD105/CD150, respectively. However, the remaining CMP population is still highly heterogenic. Thus, further separation of functional subsets within the CMP compartment is required. By screening cell surface markers that can further separate CMPs, we have identified CD27 as a reliable marker to separate all megakaryocyte/erythrocyte-committed progenitors from granulocyte/monocyte-committed progenitors. In addition, we found that CD62L is only expressed on granulocyte/monocyte-committed progenitors. CD27 and CD62L co-staining can separate CMP into CD27 +CD62L +, CD27 +CD62L - and CD27 -CD62L - subsets. Biology and morphology study showed that CD27 +CD62L - cells are closely associated with GMPs, whereas CD27 -CD62L - cells are closely associated with MEPs. In vitro culture and in vivo transplantation functional studies demonstrated that 1) CD27 +CD62L + cells are pre-GMPs that give rise to FcGRII/III + GMPs and only produce granulocytes and monocytes; 2) CD27 -CD62L - cells are pre-MEPs that give rise to MEPs and primarily produce erythrocytes and megakaryocytes with minimal contribution to granulocytes and monocytes; 3) CD27 +CD62L - subset enriches cells with genuine CMP potential capable of producing GMPs, MEPs, and subsequent progeny. Taken together, we have identified two novel populations of committed progenitors that serve as intermediates between CMP-GMP and CMP-MEP commitment pathways. Identification of pre-GMPs and pre-MEPs fills in the gap between CMPs-GMPs and CMPs-MEPs, supporting the hierarchal relationship of myeloid lineage differentiation. Disclosures No relevant conflicts of interest to declare.