SARS-CoV-2 viral clearance during bone marrow aplasia after allogeneic hematopoietic stem cell transplantation-A case report.

Respiratory viral infections are known causes of mortality after allogeneic hematopoietic stem cell transplantation (HSCT). Here, we report a unique case of a child with viral pneumonia caused by coinfection with human metapneumovirus (MPV), respiratory syncytial virus (RSV), and SARS-CoV-2 after HSCT. A 9-year-old girl with acute lymphoblastic leukemia underwent allogeneic HSCT from a matched, unrelated donor. During the post-transplant period, in profound leukopenia (below 10 leukocytes/µL), she was diagnosed with SARS-CoV-2, MPV, and RSV pneumonia and was treated with ribavirin and chloroquine. Before leukocyte recovery, the girl became asymptomatic, and SARS-CoV-2 and RSV clearance was achieved. The shedding of SARS-CoV-2 stopped before immune system recovery, and one may hypothesize that the lack of an inflammatory response might have been a contributing factor to the mild clinical course. Post-transplant care in HSCT recipients with COVID-19 infection is feasible in regular transplant units, provided the patient does not present with respiratory failure. Early and repeated testing for SARS-CoV-2 in post-transplant patients with concomitant infection mitigation strategies should be considered in children after HSCT who develop fever, respiratory symptoms, and perhaps gastrointestinal symptoms to control the spread of COVID-19 both in patients and in healthcare workers in hospital environments. Training of staff and the availability of personal protective equipment are crucial for containing SARS-CoV-2 infection.

The Kinetics of Inflammation-Related Proteins and Cytokines in Children Undergoing CAR-T Cell Therapy-Are They Biomarkers of Therapy-Related Toxicities?

CD19-targeted CAR-T cell therapy has revolutionized the treatment of relapsed/refractory (r/r) pre-B acute lymphoblastic leukemia (ALL). However, it can be associated with acute toxicities related to immune activation, particularly cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). Cytokines released from activated immune cells play a key role in their pathophysiology. This study was a prospective analysis of proinflammatory proteins and cytokines in children treated with tisagenlecleucel. Serial measurements of C-reactive protein, fibrinogen, ferritin, IL-6, IL-8, IL-10, IFNγ, and TNFα were taken before treatment and on consecutive days after infusion. The incidence of CRS was 77.8%, and the incidence of ICANS was 11.1%. No CRS of grade ≥ 3 was observed. All complications occurred within 14 days following infusion. Higher biomarker concentrations were found in children with CRS grade ≥ 2. Their levels were correlated with disease burden and CAR-T cell dose. While cytokine release syndrome was common, most cases were mild, primarily due to low disease burden before lymphodepleting chemotherapy (LDC). ICANS occurred less frequently but exhibited various clinical courses. None of the toxicities were fatal. All of the analyzed biomarkers rose within 14 days after CAR-T infusion, with most reaching their maximum around the third day following the procedure.

Blinatumomab Prior to CAR-T Cell Therapy-A Treatment Option Worth Consideration for High Disease Burden.

The optimal bridging therapy before CAR-T cell infusion in pediatric relapsed or refractory B-cell precursor acute lymphoblastic leukemia (r/r BCP-ALL) still remains an open question. The administration of blinatumomab prior to CAR-T therapy is controversial since a potential loss of CD19+ target cells may negatively impact the activation, persistence, and, as a consequence, the efficacy of subsequently used CAR-T cells. Here, we report a single-center experience in seven children with chemorefractory BCP-ALL treated with blinatumomab before CAR-T cell therapy either to reduce disease burden before apheresis (six patients) or as a bridging therapy (two patients). All patients responded to blinatumomab except one. At the time of CAR-T cell infusion, all patients were in cytological complete remission (CR). Four patients had low positive PCR-MRD, and the remaining three were MRD-negative. All patients remained in CR at day +28 after CAR-T infusion, and six out of seven patients were MRD-negative. With a median follow-up of 497 days, four patients remain in CR and MRD-negative. Three children relapsed with CD19 negative disease: two of them died, and one, who previously did not respond to blinatumomab, was successfully rescued by stem cell transplant. To conclude, blinatumomab can effectively lower disease burden with fewer side effects than standard chemotherapeutics. Therefore, it may be a valid option for patients with high-disease burden prior to CAR-T cell therapy without clear evidence of compromising efficacy; however, further investigations are necessary.

Fludarabine-Cyclophosphamide-Based Conditioning with Antithymocyte Globulin Serotherapy Is Associated with Durable Engraftment and Manageable Infections in Children with Severe Aplastic Anemia.

Severe aplastic anemia (SAA) is a bone marrow failure syndrome that can be treated with hematopoietic cell transplantation (HCT) or immunosuppressive (IS) therapy. A retrospective cohort of 56 children with SAA undergoing transplantation with fludarabine-cyclophosphamide-ATG-based conditioning (FluCyATG) was analyzed. The endpoints were overall survival (OS), event-free survival (EFS), cumulative incidence (CI) of graft versus host disease (GVHD) and CI of viral replication. Engraftment was achieved in 53/56 patients, and four patients died (two due to fungal infection, and two of neuroinfection). The median time to neutrophil engraftment was 14 days and to platelet engraftment was 16 days, and median donor chimerism was above 98%. The overall incidence of acute GVHD was 41.5%, and that of grade III-IV acute GVHD was 14.3%. Chronic GVHD was diagnosed in 14.2% of children. The probability of 2-year GVHD-free survival was 76.1%. In the univariate analysis, a higher dose of cyclophosphamide and previous IS therapy were significant risk factors for worse overall survival. Episodes of viral replication occurred in 33/56 (58.9%) patients, but did not influence OS. The main advantages of FluCyATG include early engraftment with a very high level of donor chimerism, high overall survival and a low risk of viral replication after HCT.

Non-nutritional "paramedical" usage of human milk - knowledge and opinion of breastfeeding mothers in Poland.

OBJECTIVES: The objective of this study is to understand knowledge, attitudes and practices of non-nutritional breast milk use among lactating women in respect of skin diseases and other frequent ailments. MATERIAL AND METHODS: The study, in the form of a questionnaire, spread on social media, was targeted at breastfeeding women. The questionnaire consisted of questions regarding the knowledge of non-nutritional usage of human milk, its use in practice, subjective opinion on the observed results and inclination towards future use. Chi-square tests and c-Pearson coefficients were used for statistical calculations. RESULTS: A total of 1187 responses were acted upon. In the study group, 879 women claimed to have knowledge of non-nutritional use of human milk in respect of skin and most common ailments, whilst 688 of them claimed to use at least one usage. The most frequently, breast milk was used for: care of cracked nipples, care of healthy skin, treatment of diaper dermatitis and treatment of neonatal acne. A correlation between duration of breastfeeding (p < 0.05) and gestational age (p < 0.05) and practical use of non-nutritional human milk was found. CONCLUSIONS: The study showed a great enthusiasm of mothers in respect of using breast milk for non-nutritional purposes, including the treatment of skin diseases and other common ailments. However, given the scant studies determining possible concerns surrounding these methods, there is a requirement for parental education with emphasis on the need for prompt medical examination and pertinent treatment.

Introduction of new pediatric EBMT criteria for VOD diagnosis: is it time-saving or money-wasting? : Prospective evaluation of pediatric EBMT criteria for VOD.

Hepatic veno-occlusive disease (VOD) is a potentially fatal complication following hematopoietic stem cell transplantation (HSCT). We evaluated in prospective analysis the usefulness of the pediatric EBMT criteria for VOD diagnosis and their presumable impact on cost effectiveness and patients' outcome. Study included all 282 HSCT procedures performed in Department of Pediatric Hematology/Oncology and BMT in Wroclaw between January 2016 and March 2019. Data were compared with previous VOD research conducted in our center before year 2016. Twenty-five (8.9%) patients (median age 3.5 years) were diagnosed with VOD. Duration of defibrotide (DF) administration varied from 4 to 34 days (median: 16.5), with 96% response rate. Overall survival was 88%. If applying Baltimore and modified Seattle criteria, VOD incidence was 2.13% and 5.7%, respectively. Median diagnosis delay based on modified Seattle criteria was 3 days. Before 2016, VOD incidence was 4.9%, with 74% DF response rate (p = 0.033) and 56.2% OS (p = 0.008). After implementing new criteria length of hospitalization for VOD patients decreased by median of 12 days (p = 0.009). Earlier VOD diagnosis, facilitated by EBMT criteria, resulting in implementing immediate treatment significantly improved patients' outcome. Furthermore, it allows shortening of DF administration and minimizes length of hospital stay.

Successful Bone Marrow Recovery After an Immunoablative Regimen With Autologous Cord Blood Transplant in a Child With Idiopathic Severe Aplastic Anemia: A Case Report.

BACKGROUND: Aplastic anemia is a rare disease that manifests as bone marrow failure. The current treatment options include immunoablative therapy or allogeneic hematopoietic stem cell transplantation. We report a successful immunoablative regimen with autologous umbilical cord blood (auto-UCB) transplant in a 3-year-old boy with severe aplastic anemia. CASE REPORT: The immunoablation procedure consisted of 5 × 3.75 mg/kg antithymocyte globulin (Thymoglobulin) (total 18.75 mg/kg), methylprednisolone for 4 days, and cyclosporine A. The patient received auto-UCB containing 0.3 × 105 CD34+ cells per kilogram of body weight. Recovery of leukocyte count above 1000/µL was reached on post-transplant day +39, and recovery of granulocytes above 500/µL was reached on day +40. The final regular transfusions of packed red blood cells and platelet concentrate were performed on day +55. The complications that occurred in the post-transplant period were nausea, diarrhea, septic fever, and hepatic abscess formation. Post-transplant immunosuppression with cyclosporine A was discontinued 17.5 months after auto-UCB, and the patient remained in complete remission with normal blood counts and bone marrow morphology. SUMMARY: Auto-UCB transplantation without chemotherapy conditioning can be considered a therapeutic option for children with stored cord blood cells.

Non-Nutritional Use of Human Milk Part 1: A Survey of the Use of Breast Milk as a Therapy for Mucosal Infections of Various Types in Poland.

The use of home remedies for the treatment of moderately severe ailments is a common practice in the Polish population. Currently, the topic of the potential non-nutritional properties of human milk is attracting the attention of breastfeeding mothers. This study was aimed at understanding lactating women's knowledge, attitudes, and practices of non-nutritional breast milk on mucous membranes. The study was conducted among lactating women, who filled out a questionnaire consisting of questions about their knowledge and experiences with non-nutritional use of human milk. Statistical calculations were conducted with chi-square test and c-Pearson coefficient. A total of 1187 women were acted on, whereby 768 of respondents claimed to have knowledge of the non-nutritional use of human milk on mucous membranes, whilst 404 of them claimed that they had used at least one method. Among the most frequently used methods were the treatment of rhinorrhea, lacrimal canaliculi obstruction, and conjunctivitis. A correlation between length of breastfeeding (p < 0.001) and knowledge of non-nutritional human milk usage in prophylaxis and treatment of mucous membrane inflammation was found. Breastfeeding duration (p < 0.001) and parity (p < 0.005) were correlated with the application of those methods in practice. Due to a high propensity to testing those methods, parents' education in the field of possible risks and importance of medical consultations is necessary.

Prognostic impact of combined fludarabine, treosulfan and mitoxantrone resistance profile in childhood acute myeloid leukemia.

BACKGROUND: The role of cellular drug resistance in childhood acute myeloid leukemia (AML) has not yet been established. The aim of the study was the analysis of the clinical value of ex vivo drug resistance in pediatric AML. PATIENTS AND METHODS: A cohort of 90 children with de novo AML were assayed for drug resistance profile by the 3-4,5-dimethylthiazol-2-yl-2,5-difenyl tetrazolium bromide (MTT) assay and prognostic model of in vitro drug sensitivity was analyzed. RESULTS: Children who relapsed during follow-up showed higher in vitro resistance of leukemic blasts to most of the drugs tested, except for cytarabine, cladribine, vincristine, mercaptopurine and thioguanine. A combined in vitro drug resistance profile to fludarabine, treosulfan and mitoxantrone (FTM score) was defined and it had an independent prognostic significance for disease free survival in pediatric AML. CONCLUSION: The combined fludarabine, treosulfan and mitoxantrone resistance profile to possibly may be used for better stratification of children with AML or indicate the necessity for additional therapy.

Predictive value of multidrug resistance proteins and cellular drug resistance in childhood relapsed acute lymphoblastic leukemia.

PURPOSE: Cellular resistance in childhood acute leukemias might be related to profile and function of multidrug resistance proteins and apoptosis regulating proteins. The aims of the study were: (1) analysis of expression of MRP1, PGP1, LRP, BCL-2 and p53 proteins; (2) correlation with ex vivo drug resistance, and (3) analysis of their prognostic impact on clinical outcome in childhood acute lymphoblastic (ALL) and acute myeloid (AML) leukemia. METHODS: Total number of 787 children diagnosed for initial ALL (n = 527), relapsed ALL (n = 104), initial AML (n = 133) and relapsed AML (n = 23) were included into the study. Mean follow-up period was 3.5 years. Drug resistance for up to 30 anticancer agents was performed by the MTT assay. Expression of all proteins was tested by flow cytometry. RESULTS: Both initial AML and relapsed ALL samples showed higher drug resistance than initial ALL samples. No significant differences were found in drug resistance between initial and relapsed AML samples. The presence of multidrug resistance and apoptosis proteins had no impact on pDFS in iALL and iAML, however strong trend towards adverse prognostic impact of MRP1, PGP and LRP on pDFS in rALL was observed. The same trend was observed for each of analyzed co-expressions of tested multidrug resistance proteins. CONCLUSIONS: The phenomenon of cellular drug resistance in childhood acute leukemias is multifactorial and plays an important role in response to therapy. Expression of MRP1, PGP and LRP proteins, as well as their co-expression play possible role in childhood relapsed ALL.

CD40L and IL-4 stimulation of acute lymphoblastic leukemia cells results in upregulation of mRNA level of FLICE--an important component of apoptosis.

The use of cancer vaccines based on dendritic cells (DC) presenting tumor antigens can be a promising tool in the treatment of leukemia. The functional characteristics of leukemia derived DC is still to be elucidated. CD40 promotes survival, proliferation and differentiation of normal B cells. CD40 triggering was used to enhance the poor antigen-presenting capacity of leukemic B-cells. Since it is still unclear whether CD40 ligation drives neoplastic B-cells to apoptosis or not, we assessed the mRNA expression of FLICE, FAS, FADD and TRADD - important components of apoptosis machinery, using real-time PCR in acute lymphoblastic leukemia cells before and after CD40 and IL-4 stimulation. ALL cells stimulated with CD40L/IL-4 expressed dendritic cell phenotype at mRNA and protein levels (upregulation of main costimulatory and adhesion molecules noted in real-time RT PCR and flow cytometry); they also expressed higher amounts of mRNA for FLICE, TRADD and FADD after CD40L/IL-4 stimulation. However differences statistically significant comparing cells cultured with CD40L/IL-4 and medium alone regarded only FLICE. Concluding, we showed upregulation of important elements of apoptosis at mRNA level in ALL cells after CD40 ligation.

[Developmental anomaly of superior vena cava as a reason hampering vein-port catheter implantation]. / Nieprawidlowy splyw ukladu zyly glównej górnej jako przyczyna uniemozliwiajaca wszczepienie portu naczyniowego.

Occasionally unexpected technical difficulties occur during Port-A-Cath implantation and the central venous catheterization. We described a case of superior vena cava developmental anomaly diagnosed during Port-A-Cath implantation.

[Amniotic fluid analysis. Part I: Rapid markers in the prediction of intra-amniotic infection]. / Diagnostyczne badania laboratoryjne plynu owodniowego. Czesc I: Szybkie testy w przewidywaniu zakazenia wewnatrzowodniowego.

Intraamniotic infection has been recognized as a major etiologic factor for preterm delivery. Several groups have proposed that amniocentesis be used to identify the patient at risk for infectious morbidity. The number of techniques have been studied for rapid identification of bacterial colonization of amniotic cavity. Diagnostic index value of Gram stain, white blood cell count, glucose level and LDH (lactate dehydrogenase) activity for prediction of positive amniotic fluid culture, preterm delivery, clinical infection and neonatal sepsis were shown in the study. Investigators continue attempts to establish a rapid, more useful tests to predict preterm delivery.

Individualized tumor response testing profile has a prognostic value in childhood acute leukemias: multicenter non-interventional long-term follow-up study.

A total number of 817 children with acute lymphoblastic leukemia (ALL) and 181 with acute myeloblastic leukemia (AML) were assessed for individualized tumor response testing (ITRT) profile as a prognostic factor in long-term follow-up. For each patient, ITRT, initial response to therapy and long-term outcome were assessed. In initial ALL, an impact on long-term response was shown in ITRT for 13 drugs, while in initial AML only for cytarabine. For patients with ALL, a combined five-drug ITRT profile for prednisolone, l-asparaginase, vincristine, cytarabine and daunorubicin or doxorubicin had predictive value for probability of disease-free survival (pDFS) in univariate analysis, whereas in multivariate analysis, bone marrow response by day 33 was the only prognostic factor. For patients with AML, no factor had prognostic value for pDFS in univariate analysis, while ITRT to cytarabine almost reached significance. In conclusion, ITRT can possibly be regarded as a risk factor in childhood acute leukemias.

Late-preterm cesarean delivery and chemokines concentration in the umbilical cord blood of neonates.

OBJECTIVE: The objective of the study was to investigate whether concentrations of chemokines in the umbilical cord blood of neonates are affected by delivery via cesarean section. STUDY DESIGN: Umbilical cord blood was obtained from 116 singleton late-preterm and full-term neonates without infections, born to healthy pregnant women. Concentrations of chemokines - MIP-1α (CCL3), MIP-1ß 1 (CCL4), RANTES (CCL5), GRO-α (CXCL1) and ENA-78 (CXCL5) - were measured by ELISA. Logistic regression was used to investigate regression relationships between the occurrence of neonatal chemokines concentrations in umbilical cord blood and mode and time of delivery. RESULTS: Concentrations of CXC chemokines in late-preterm neonates were the same as those in term neonates. RANTES concentrations in late-preterm cord blood were significantly lower than concentrations in term cord blood. Concentrations of the CC chemokine - RANTES (CCL5) - were noted to be lower in neonates born to cesarean section than in neonates born vaginally. Any anesthetic taken by the mothers during cesarean section did not affect CC chemokine production in the cord blood of full-term neonates. In a logistic regression model including gestational age as a variable, late-preterm delivery was associated with RANTES concentrations (OR = 3.8). After adjustment for variable mode of delivery in regression model, RANTES concentration (OR = 4.75). CONCLUSION: Both late-preterm and cesarean delivery are essential risk factors of low RANTES (CCL5) concentrations in the umbilical cord blood.

Differential ex vivo activity of bortezomib in newly diagnosed paediatric acute lymphoblastic and myeloblastic leukaemia.

The proteasome inhibitor, bortezomib, is known to be effective in the therapy of various neoplasms. The objective of this study was the analysis of the ex vivo activity of bortezomib in paediatric acute lymphoblastic leukaemia (ALL), in comparison to paediatric acute myeloid leukaemia (AML). A total of 159 patients entered the study, including 106 ALL (including 86 precursor-B-cell ALL, and 20 T-cell ALL) and 53 AML children. The ex vivo sensitivity to bortezomib and 16 other drugs was studied by MTT assay. Paediatric AML samples were more resistant than paediatric ALL samples to most of the tested drugs, except for cytarabine and thioguanine. With respect to immunophenotype, ex vivo drug resistance in T-cell ALL (T-ALL) was higher for most of the drugs. No differences in drug resistance between T-ALL and common/pre-B-cell-ALL were found for daunorubicin, mitoxantrone and 6-thioguanine. Bortezomib was the only compound which was more active in T-ALL than in common/pre-B-ALL paediatric samples. In conclusion, bortezomib had good ex vivo activity in paediatric T-ALL samples.

Umbilical serum concentrations of chemokines (RANTES and MGSA/GRO-alpha) in preterm and term neonates.

BACKGROUND: The objective of this study was to explore the relationship between labor (preterm and term) and umbilical blood serum regulated on activation, normal T cell expressed and secreted (RANTES) and melanoma growth stimulatory activity/growth-related oncogene-a (MSGA/GRO-alpha) concentration, and to determine whether early sepsis and pneumonia are associated with changes in concentrations of the chemokines (RANTES and MSGA/GRO-alpha) in umbilical blood serum. METHODS: Umbilical blood was obtained from 67 neonates in the following groups: (i) preterm neonates with early sepsis; (ii) preterm neonates with pneumonia; (iii) non-infected preterm neonates; and (iv) full-term healthy neonates. RANTES and MGSA/GRO-alpha concentrations were determined by use of a commercially available immunoassay kit. RESULTS: Non-infected preterm neonates had lower RANTES concentrations than healthy term neonates. Preterm infected neonates (pneumonia or sepsis) did not have higher RANTES concentrations than non-infected preterm neonates. In contrast, non-infected preterm neonates had higher MSGA/GRO-alpha concentrations than full-term healthy neonates. And preterm neonates with sepsis had higher MGSA/GRO-alpha concentrations than preterm ones with pneumonia and non-infected preterm ones. CONCLUSIONS: Preterm neonates had constitutively lower RANTES concentrations than term ones and it seems that during infection RANTES concentrations did not increase. MGSA/GRO-a concentrations were constitutively higher in preterm than term neonates, and septic events further increased its concentrations in preterm neonates.

Assessment of chosen parameters of the immune system in children with acute lymphoblastic leukemia.

The immunosuppressive effect of cytostatics, basic therapeutic agents in the treatment of proliferative diseases of the hematopoietic system, and the rising number of children cured from acute leukaemias form together a need to monitor the status of the immune system following cessation of therapy. Surface antigens in lymphocytes from peripheral blood were assessed in 16 children directly after intensive chemotherapy (i.e., after protocol II of the BMF program) and in 25 children 12-13 months following conclusion of acute lymphoblastic leukemia treatment. The results were compared to data obtained from children never afflicted with neoplastic disease. A significant decrease in the average number of lymphocyte subpopulations was noted in the case of the treated children directly after intensive chemotherapy. The average values of lymphocyte subpopulations in children with concluded treatment are within the norm, with the exception of NK and TS lymphocytes.