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Background: Botulinum toxin A (BoNT‐A) is widely utilized in the management of hypertrophic and keloid scars. One proposed mechanism for scar prevention involves the inhibition of fibroblast migration in scars by BoNT‐A. However, the data regarding the effect of BoNT‐A on the migration of normal human dermal fibroblasts (NHDF) is limited. Objectives: The aim of this study was to investigate the inhibitory effect of different types and dilutions of BoNT‐A on the migration of NHDF. Methods: In vitro scratch wound assay, NHDF cells were cultured, incubated, and subjected to scratching using a sterile tip. Subsequently, the scratched NHDF monolayer was treated with different types of BoNT‐A, including onabotulinumtoxinA (ONA), incobotulinumtoxinA (INCO), prabotulinumtoxinA (PRABO), or letibotulinumtoxinA (LETI), at varying concentrations of 10, 20, 25, 40, 50, and 100 units/milliliter (U/mL). Additionally, abobotulinumtoxinA (ABO) was administered at concentrations of 33, 50, 66, 71, 100, 150, 300, and 500 U/mL. Normal saline solution (NSS) served as a negative control. The extent of NHDF migration was evaluated by comparing each dilution of BoNT‐A with the controls using high‐content imaging at the 48‐h time point. Furthermore, the viability of the of NHDF was assessed. Results: The concentrations of 25, 40, and 50 U/mL of ONA (p < 0.001) and 25 U/mL of LETI (p < 0.05) demonstrated significantly inhibited NHDF migration in comparison to the control group. Conversely, all dilutions of PRABO, INCO, and ABO exhibited comparable NHDF migration to that of the control group. Regarding NHDF viability, no significant decrease was observed across any of the BoNT‐A types and dilutions. Conclusion: Different types and dilutions of BoNT‐A demonstrated variable inhibitory effects on NHDF migration in vitro. The selection of BoNT‐A formulation may significantly impact the clinical outcome of scar prevention related to fibroblast migration. [ABSTRACT FROM AUTHOR]

MYCN proto‐oncogene, bHLH transcription factor (MYCN) amplification is associated with aggressive retinoblastoma (RB) and neuroblastoma (NB) cancer recurrence that is resistant to chemotherapies. Therefore, there is an urgent need to identify new therapeutic tools. This study aimed to evaluate the potential repurposing of ceftriaxone for the treatment of MYCN‐amplified RB and NB, based on the clinical observations that the drug was serendipitously found to decrease the volume of the MYCN‐driven RB subtype. Using patient‐derived tumor organoids and tumor cell lines, we demonstrated that ceftriaxone is a potent and selective growth inhibitor targeting MYCN‐driven RB and NB cells. Profiling of drug‐induced transcriptomic changes, cell‐cycle progression, and apoptotic death indicated cell‐cycle arrest and death of drug‐treated MYCN‐amplified tumor cells. Drug target identification, using an affinity‐based proteomic and molecular docking approach, and functional studies of the target proteins revealed that ceftriaxone targeted DEAD‐box helicase 3 X‐linked (DDX3X), thereby inhibiting translation in MYCN‐amplified tumors but not in MYCN‐nonamplified cells. The data suggest the feasibility of repurposing ceftriaxone as an anticancer drug and provide insights into the mechanism of drug action, highlighting DDX3X as a potential target for treating MYCN‐driven tumors. [ABSTRACT FROM AUTHOR]

Objectives: Multiple myeloma (MM) accounts for 10% of hematologic malignancies. However, most of the patients suffered from relapsed/refractory disease. We would like to expand CAR T cell therapy to treat MM using our current platform. Methods: BCMA CAR T lymphocytes were generated for volunteers or MM patients. The transduction efficiency was detected by the ddPCR technique. Immunophenotyping and exhaustion markers were monitored by flow cytometry. The efficacy of BCMA CAR T cells was tested using coculturing with BCMA CAR or mock, and the positive and negative targets, K562/hBCMA‐ECTM and K562, respectively. Results: BCMA CAR T cells were generated from consented volunteers or MM patients and could be detected CAR BCMA expression at a mean of 4.07 ± 1.95 or 4.65 ± 1.21 copies/cell, respectively. Those modified T cells were primarily effector memory T cells. Our BCMA CAR T cells could explicitly eradicate the K562/hBCMA‐ECTM cell line while the K562 cell line survived. Interestingly, the BCMA CAR, mock T cells, and peripheral blood mononuclear cells from MM patients expressed similar levels of the exhaustion makers, TIM‐3, LAG‐3, and PD1. Conclusions: Our BCMA CAR T cells, mainly effector/effector memory, could eliminate BCMA‐expressing cells in vitro and had similar levels of exhaustion markers among different populations. [ABSTRACT FROM AUTHOR]

Introduction: BK polyomavirus (BKPyV)‐associated nephropathy (BKPyVAN) can cause a significant risk of allograft impairment after kidney transplantation (KT). Intact BKPyV‐specific immunity is associated with viral containment. This study investigated BKPyV‐specific immunological factors among KT recipients. Methods: This prospective study in a single transplant center from January 2019 to August 2019 assessed associations between clinical and immunological characteristics, with a focus on BKPyV‐cell‐specific immunity and BKPyVAN, among KT recipients aged ≥15 years. The numbers of interferon‐gamma (IFN‐γ)‐producing CD4+ T, CD8+ T, natural killer (NK), and natural killer T (NKT) cells were measured after stimulation with large T antigen and viral capsid protein 1 (VP1). Results: In total, 100 KT recipients were included (mean age ± SD, 42 ± 11 years); 35% of the recipients were female patients, and 70% had received induction immunosuppressive therapy. The 1‐year cumulative incidence of high‐level BKPyV DNAuria (possible BKPyVAN) and (presumptive BKPyVAN) was 18%. Among 40 patients with immunological factor data, pre‐KT %NK cells (hazard ratio [HR], 1.258; 95% confidence interval [CI], 1.077–1.469; p =.004) and %VP1‐specific NK cells (HR, 1.209; 95% CI, 1.055–1.386; p =.006) were factors independently associated with possible and presumptive BKPyVAN. KT recipients with possible and presumptive BKPyVAN were more likely to exhibit significant mean coefficients of %NK, %VP1‐specific NK, and %NKT cells at 1 month after KT than before KT (all p <.05). Conclusion: Individuals with nonspecific and VP1‐specific NK cells before KT and increasing numbers of these cells after KT may be at risk for high‐level BKPyV DNAuria and presumptive BKPyVAN. Further studies are needed to determine the utility of BKPyV‐specific innate immune surveillance in predicting the occurrence of BKPyVAN. [ABSTRACT FROM AUTHOR]

Anticancer peptides (ACPs) are rising as a new strategy for cancer therapy. However, traditional laboratory screening to find and identify novel ACPs from hundreds to thousands of peptides is costly and time consuming. Here, a sequential procedure is applied to identify candidate ACPs from a computer‐generated peptide library inspired by alpha‐lactalbumin, a milk protein with known anticancer properties. A total of 2688 distinct peptides, 5–25 amino acids in length, are generated from alpha‐lactalbumin. In silico ACP screening using the physicochemical and structural filters and three machine learning models lead to the top candidate peptides ALA‐A1 and ALA‐A2. In vitro screening against five human cancer cell lines supports ALA‐A2 as the positive hit. ALA‐A2 selectively kills A549 lung cancer cells in a dose‐dependent manner, with no hemolytic side effects, and acts as a cell penetrating peptide without membranolytic effects. Sequential window acquisition of all theorical fragment ions‐proteomics and functional validation reveal that ALA‐A2 induces autophagy to mediate lung cancer cell death. This approach to identify ALA‐A2 is time and cost‐effective. Further investigations are warranted to elucidate the exact intracellular targets of ALA‐A2. Moreover, these findings support the use of larger computational peptide libraries built upon multiple proteins to further advance ACP research and development. [ABSTRACT FROM AUTHOR]

Objectives: Patients with high‐risk hematologic diseases require intensive modalities, including high‐dose chemotherapy and allogeneic hematopoietic stem cell transplantation (allo‐HSCT). Haploidentical T‐cell–replete transplantation is a logical choice because of the limited availability of matched sibling donors and the prolonged time needed to identify matched unrelated donors in Thailand. Methods: The clinical outcomes data of 43 patients undergoing allo‐HSCT were reviewed. All patients had high‐risk hematologic malignancies, were younger than 20 years, and were in complete cytological remission at the time of allo‐HSCT. We used two different conditioning regimens: total body irradiation (TBI) combined with cyclophosphamide, fludarabine, and melphalan (n = 23) and thiotepa combined with fludarabine and busulfan (n = 20). All patients received a graft‐versus‐host disease prophylaxis regimen consisting of cyclophosphamide, mycophenolate mofetil, and a calcineurin inhibitor or sirolimus. Results: There was no difference in engraftment between patients receiving either of the regimens. After a median follow‐up of 35.8 (range, 0.6–106.2) months, the overall survival (OS) and event‐free survival (EFS) rates were 62.4% and 54.7%, respectively. OS and EFS were comparable between the respective regimens. Conclusions: We conclude that thiotepa‐based conditioning has similar efficacy and tolerability as TBI‐based conditioning for haploidentical HSCT with post‐transplant cyclophosphamide. [ABSTRACT FROM AUTHOR]

The interaction of SARS‐CoV‐2 infection with extracellular vesicles (EVs) is of particular interest at the moment. Studying SARS‐CoV‐2 contaminated‐EV isolates in instruments located outside of the biosafety level‐3 (BSL‐3) environment requires knowing how viral inactivation methods affect the structure and function of extracellular vesicles (EVs). Therefore, three common viral inactivation methods, ultraviolet‐C (UVC; 1350 mJ/cm2), β‐propiolactone (BPL; 0.005%), heat (56°C, 45 min) were performed on defined EV particles and their proteins, RNAs, and function. Small EVs were isolated from the supernatant of SARS‐CoV‐2‐infected human lung epithelial Calu‐3 cells by stepwise centrifugation, ultrafiltration and qEV size‐exclusion chromatography. The EV isolates contained SARS‐CoV‐2. UVC, BPL and heat completely abolished SARS‐CoV‐2 infectivity of the contaminated EVs. Particle detection by electron microscopy and nanoparticle tracking was less affected by UVC and BPL than heat treatment. Western blot analysis of EV markers was not affected by any of these three methods. UVC reduced SARS‐CoV‐2 spike detectability by quantitative RT‐PCR and slightly altered EV‐derived β‐actin detection. Fibroblast migration‐wound healing activity of the SARS‐CoV‐2 contaminated‐EV isolate was only retained after UVC treatment. In conclusion, specific viral inactivation methods are compatible with specific measures in SARS‐CoV‐2 contaminated‐EV isolates. UVC treatment seems preferable for studying functions of EVs released from SARS‐CoV‐2 infected cells. [ABSTRACT FROM AUTHOR]

Anti-SARS-CoV-2 effect of extracellular vesicles released from mesenchymal stem cells As of 10 December 2021, coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 accounted for 267 million people with up to 5.3 million deaths worldwide (https://covid19.who.int). Accordingly, we applied in vitro anti-SARS-CoV-2 assays, which were previously developed by our group (Kanjanasirirat et al., 2020; Kongsomros et al., 2021; Sa-Ngiamsuntorn et al., 2021), to determine the dose dependent anti-SARS-CoV-2 effect of MSC-EVs. In this direction, MSC-EV investigations for COVID-19 treatment would be more appealing and undeniable if MSC-EVs also exhibit anti-SARS-CoV-2 effects. [Extracted from the article]

Aims: The purpose of this study was to design and manufacture CD19 chimeric antigen receptor (CAR)‐modified T cells for clinical use in Thailand, as a model for how this technology can be directly applied at individual institutions treating high‐risk leukemia patients. Methods: We constructed second‐generation CAR T cells expressing CD19 scFV‐CD28‐CD3ζ with different lengths of the spacer region: full, intermediate, and short length, by using a lentiviral vector. We wanted to determine whether the difference in length of the spacer would affect the cytotoxic potential of the CD19 CAR T cells against the leukemic cells. Results: We found that all constructs of CD19 CAR T cells exhibited a similar level of cytotoxicity against several human lymphoma and leukemia cell lines. For the clinical application, we chose the intermediate length spacer construct CD19 CAR T cells, hypothesizing that the highest transduction efficiency coupled with a slower initial proliferation in vitro might lead to effective leukemic cell kill, yet a lower probability for serious clinical side effects. We then tested the clinical efficacy of our CD19 CAR T cells in one patient with refractory/relapsed acute B‐cell lymphoblastic leukemia. This patient indeed had minimal clinical side effects after the CAR T‐cell infusion, and he remains in an unmaintained, ongoing complete remission 10+ months after his T‐cell treatment. Conclusion: Our CD19 CAR T cells demonstrated efficacies in acute lymphoblastic B‐cell leukemia, and will be used to establish an immunotherapeutic program for high‐risk B‐cell acute lymphoblastic leukemia in Thailand. We propose that this approach can be used as a model for how this new exciting technology can be applied directly at individual institutions that treat (a large number of) patients with high‐risk leukemia. [ABSTRACT FROM AUTHOR]

Keywords: fetal haemoglobin induction; -thalassaemia/HbE; pomalidomide; hydroxyurea; decitabine EN fetal haemoglobin induction -thalassaemia/HbE pomalidomide hydroxyurea decitabine e240 e245 6 06/18/20 20200615 NES 200615 Studies have shown that increased expression of fetal haemoglobin (HbF; SB 2 sb SB 2 sb ) can ameliorate red blood cell deficiencies in patients with -thalassaemia and sickle cell disease (SCD).[[1], [3]] Pharmacological induction of HbF expression in -thalassaemia has been investigated using several classes of small molecules,[4] including 5-azacytidine,[5] decitabine,[6] hydroxyurea,[7] LSD1 inhibitors (tranylcypromine and RN-1),[[8]] and short chain fatty acid derivatives.[[10]] Among these molecules, hydroxyurea is the only U.S. Food and Drug Administration (FDA) currently approved drug for the treatment of SCD and/or -thalassaemia. SP 0 sp -thalassaemia/HbE precursors from patients of different SP 0 sp -thalassemic mutations (Table SI) showed similarly increased levels of HbF induction in response to pomalidomide treatment. GLO:1XW/15jun20:bjh16670-fig-0002.jpg PHOTO (COLOR): 2 Effect of pomalidomide and its combinations on erythroid differentiation and mRNA expression of HbF regulators in cultured erythroid cells from 0-thalassaemia/HbE patients. HbF-inducing effects of hydroxyurea (HU), decitabine (DAC), and RN-1 in erythroid cells from 0-thalassaemia/HbE patients. [Extracted from the article]

Introduction: Human adenovirus (HAdV) infection can cause substantial morbidity in kidney transplant (KT) recipients. Cell‐mediated immunity plays an important role in controlling HAdV infection after KT. Methods: We prospectively (January 2015 to June 2018) investigated the absolute lymphocyte count (ALC) and interferon‐γ‐producing CD4+ and CD8 + T cells at diagnosis and at viral clearance by an intracellular cytokine assay after stimulating with HAdV whole lysate, hexon, and penton proteins. HAdV infection was defined as the presence of HAdV DNA load in plasma or clinical specimens measured by the polymerase chain reaction assay. Results: Eighteen adult KT recipients were diagnosed with HAdV infection at a median of 16 months (interquartile range [IQR], 2‐39) after KT. The majority (94%) had HAdV‐associated hemorrhagic cystitis. The median ALC at viral clearance was significantly higher compared with diagnosis (2257 cells/mm3 [IQR, 1544‐3078] vs 1001 cells/mm3 [IQR, 641‐1385]; P < 0.001). Eleven patients underwent measurement of the HAdV‐specific T‐cell response. The median numbers of CD4+ and CD8+ T cells at viral clearance were significantly higher compared with diagnosis (448 cells/mm3 [IQR, 248‐651] vs 215 cells/mm3 [IQR, 159‐272], P = 0.02; and 623 cells/mm3 [IQR, 242‐772] vs 235 cells/mm3 [IQR, 129‐266], P < 0.01), respectively. The median percentages of penton‐specific CD4+ and hexon‐specific CD8+ T cells at viral clearance were significantly higher compared with diagnosis (0.012% vs 0%, P = 0.03%; and 0.136% vs 0.016%, P = 0.003, respectively). Conclusions: Our findings suggest a trend of ALC and HAdV‐specific T‐cell immune restoration in KT recipients who achieve successful HAdV clearance. [ABSTRACT FROM AUTHOR]

PTLD is a rare but potentially life‐threatening condition, which shows a higher prevalence in children than in adults. From 129 children who underwent LT, we reported 5 cases with biopsy‐proven PTLD at a single teaching hospital. Four patients had shared clinical presentations including fever, lymphadenopathy, and splenomegaly. They were noted to be given a prolonged course of IS due to the management of comorbid complications such as acute cellular rejection or severe food allergy or eosinophilic gastrointestinal disease. The other one patient presented with upper gastrointestinal bleeding from gastric mass during an early post‐transplantation period. Notably, hypoalbuminemia was noted in all reported patients. Similar to previous studies, both EBV serology mismatch between the donor and recipient with high EBV viral load were noted in all except one case, whose EBV serology was unknown before LT. At least one episode of CMV reactivation was also observed in 3 of 5 patients prior to the PTLD diagnosis. The histopathology revealed 1 of 5 early PTLD, 1 of 5 polymorphic PTLD, and 3 of 5 monomorphic PTLD. The treatment included IS withdrawal, chemotherapy, and/or rituximab. One patient died of multiorgan dysfunction, one remains in complete remission, and three patients are either still on treatment or await response evaluation. Even though most of our reported PTLD cases had shared manifestations with fever, lymphadenopathy, splenomegaly, EBV serology mismatch, and high EBV viral load, various initial presentations such as respiratory symptoms, hypoalbuminemia, and prolonged use of IS from other causes such as significant food allergy were noted. [ABSTRACT FROM AUTHOR]

Background: In the modern era of chemotherapy, the outcome of pediatric non‐Hodgkin lymphoma (NHL) continues to improve internationally. Limited data such as information on epidemiology and survival, however, are available in Asian countries. Methods: Children (≤15 years old) diagnosed with histologically proven NHL from 1998 to 2014 were retrospectively analyzed. Results: In total, 114 patients were enrolled; they were predominantly male (65.8%) and had advanced disease (stage III, IV; 71.9%). Of these, 22.8% had Burkitt lymphoma, 20.2% had diffuse large B‐cell lymphoma, 21.1% had lymphoblastic lymphoma, 20.2% had large cell lymphoma, and 15.8% had peripheral T‐cell lymphoma. Twenty‐nine patients died, especially of uncontrolled disease (62.1%) and infection (20.7%). During a median follow up of 78.4 months, Kaplan–Meier 5 year event‐free and overall survival rates were 71.5% ± 4.3% and 74.8% ± 4.1%, respectively, regardless of subtype. B symptoms (i.e. systemic symptoms of fever, night sweats, and weight loss that can be associated with both Hodgkin's lymphoma and non‐Hodgkin's lymphoma) and advanced disease had a significant negative impact on 5 year survival. No other prognostic factor was found, but survival tended to have a negative correlation with age. Conclusions: Pediatric NHL is aggressive, with a high prevalence of peripheral T‐cell lymphoma. The present treatment stratification seems to be effective compared with that used in developed countries. [ABSTRACT FROM AUTHOR]

Owing to the rarity of DKA as compared with the other causes of headache in this particular patient, the DKA diagnosis was missed. Intracranial hypertension could be a presenting symptom of diabetic ketoacidosis. Acetazolamide aggravated diabetic ketoacidosis severity in a boy post-transplantation thalassaemia with intracranial hypertension. [Extracted from the article]

Viral hemorrhagic cystitis ( HC) after hematopoietic stem cell transplantation ( HSCT) can be devastating. Standard treatment modalities have not been well established, but immune reconstitution may be necessary for sustained viral clearance. We studied five pediatric patients who developed viral HC after haplo-identical HSCT. All patients developed virus-specific CD4- and CD8-positive T cells, and the emergence of these viral-specific T cells was temporally associated with successful viral clearance. [ABSTRACT FROM AUTHOR]

Recently, peripheral blood siem cell (PBSC) has been widely used and replaced bone marrow (BM) as the stem cell source in allogeneic hematopoietic stem cell transplantation (HSCT) because of a more rapid engraftment, easier accessibility, and lower risk of donor complications. We, therefore, report the predicting factors for the high PBSC harvest yields in 50 healthy donors. Among the 50 donors, median collected CD34 + cell number was 4.6 × 106kg (1.5-16.3 × 106/kg). Number of circulating CD34+ cells and hematocrit (HCT) level increased parallelly whereas peripheral CD34+ cell numbers were decreased with increasing donor age. In univariate analysis, HCT level ⩾ 35.5% at the time of PBSC collection was significantly associated with high PBSC number (> 5.0 × 106 cells/kg) and donor aged <30 years was significantly associated with collected CD34+ cells>6.0 × 106/kg, P = 0.03. HCT level >35.5% was an independent parameter for high WBC count (>50 × 107L), P<0.05. None of donor who had both HCT < 35.5% and WBC < 50 × 109/L had circulating CD34+ cells⩾5.0 × 106/kg. Platelet count>200 × 109/L was found significantly in donors with WBC ⩾ 40 × 10y/L (P = 0.03) and HCT ⩾ 35.5%, P<0.05. Collected PBSC number tended to be higher in our donors with high levels of HCT, WBC, and platelet. We also found that HCT and platelet levels in our donors decreased after receiving G-CSF administration compared with the initial complete blood counts (CBC) results. We, therefore, concluded that HCT level at the time of initiation leukapheresis was an important predictor for PBSC collection yields. J. Clin. [ABSTRACT FROM AUTHOR]

The article presents a case study of a 19-year-old male adolescent with epithelioid sarcoma of lumbar vertebrae presented with fever and shock. His spinal sarcoma had been diagnosed since he was 8 years of age and was treated with surgical resection, and recurrence of a tumor with metastases had occurred since he was 12 years of age. Two weeks following the discontinuation of hydrocortisone, adrenal reserve function was evaluated, and uncommon cause of adrenal insufficiency was concluded.

We retrospectively analysed the outcomes of 127 children with acquired severe aplastic anaemia ( SAA) who had received haematopoietic stem cell transplantation ( HSCT) between 2000 and 2011 in one of the 10 Asia Pacific institutions. Fifty-three were matched sibling donor ( MSD) and 74 were alternative donor ( AD), including 22 matched unrelated donor, 32 mismatched unrelated donor and 20 mismatched related donor. With a median follow up 45·5 months (13-139) and when compared to the MSD group, AD recipients had more grade II- IV acute graft-versus-host disease (a GVHD; 14·3% vs. 32·8%, P = 0·029), but similar grade III- IV a GVHD (10·2% vs. 12·5%, P = 0·774), graft failure ( GF) (15·1% vs. 15·5%, P = 0·658) and 5-year overall survival (90·6% vs. 83·7%, P = 0·251). As a source of stem cell, peripheral blood stem cells ( PBSC) resulted in less GF (18% vs. 9·1% P = 0·013), similar grade II-IV a GVHD (28·1% vs. 17·4%, P = 0·258), chronic GVHD (25·8% vs. 29·3%, P = 0·822) and similar outcomes (89·7% vs. 82·4%, P =0 ·665) when compared to bone marrow ( BM). In univariate analysis, GF ( P < 0·001) and grade II- IV a GVHD ( P = 0·009) were predictors of poor survival. In multivariate analysis, only GF was associated with poor survival ( P = 0·012). The outcome of AD and PBSC HSCT were comparable to that of MSD and BM HSCT in the Asia Pacific region. [ABSTRACT FROM AUTHOR]

Tangnararatchakit K, Tirapanich W, Anurathapan U, Tapaneya-Olarn W, Pakakasama S, Jootar S, Slavin S, Hongeng S. Depletion of alloreactive T cells for tolerance induction in a recipient of kidney and hematopoietic stem cell transplantations. Abstract: The present case report represents a successful attempt to induce transplantation tolerance to organ allograft by combined administration of donor hematopoietic cells and kidney based on in vivo deletion of alloreactive host-vs-graft and graft-vs-host alloreactive T cells following non-myeloablative conditioning. We were able to induce mixed and eventually full donor chimerism and tolerance of kidney allograft in a 15-yr-old male with ESRD after cisplatin treatment and autologous HSCT for mediastinal germ cell tumor. Our approach to induce tolerance was based on preferential depletion of alloreactive T cells induced by exposure to donor's alloantigens and administration of cyclophosphamide at day 2 and day 3 after stem cell infusion. Additional non-specific immunosuppression as part of the conditioning included exposure to two fractions of TLI, treatment with alemtuzumab (monoclonal anti-CD52) and short-term conventional IS treatment to avoid early graft loss, because of request of IRB. Using this approach, with rapid tapering of all conventional IS treatment, the patient maintains good renal functions without evidence of both acute and chronic rejection for 32 months off all medications. [ABSTRACT FROM AUTHOR]

Kulpiya A, Mahachoklertwattana P, Pakakasama S, Hongeng S, Poomthavorn P. Hypercalcemia and altered biochemical bone markers in post-bone marrow transplantation osteopetrosis: A case report and literature review. Abstract: Autosomal recessive osteopetrosis is a rare disorder of bone resorption defect that results in generalized sclerotic bones and bone marrow failure. Allogeneic BMT is the only treatment for cure. One of the complications following a successful BMT is hypercalcemia that is a unique complication in this group of patients. We report a three-yr-old boy with osteopetrosis who developed hypercalcemia following the successful BMT. His maximal calcium level was 13.3 mg/dL. Markedly increased both bone formation and resorption markers were demonstrated along with hypercalcemia. These findings indicated an active donor-derived osteoclastic function and thus bone resorption following the successful donor engraftment in the patient. Treatment with hyperhydration, furosemide and bone resorption inhibitors, calcitonin, and bisphosphonate led to normalization of the serum calcium level. Bone resorption but not bone formation marker was persistently elevated despite having normocalcemia during a 16.5-month follow-up period. [ABSTRACT FROM AUTHOR]

Background: Patients with febrile neutropenia (FN) may develop severe infection, septic shock, and death. To improve the outcome of pediatric oncology patients with suspected FN, clinical practice guidelines were developed for these patients at the emergency room (ER). The objective of the present study was to evaluate compliance of the clinical practice guidelines for children with cancer presenting with fever to the ER and adverse outcomes after using the guidelines. Methods: A retrospective cohort study was undertaken of children with cancer presenting with fever to the ER from January 2007 to December 2008 after the clinical guidelines were implemented. The control group was the children with cancer who presented with fever during January 2005-December 2006. Guideline compliance was evaluated by recording the time of initial clinical and laboratory assessment and door-to-antibiotic time. The adverse outcomes, including septic shock and death, were determined. Results: There were 170 febrile episodes after using the guidelines. Approximately half (49.4%) of the patients received clinical assessment and laboratory results within 60 min, whereas the antibiotics were administered within 120 min in 80%. Prevalence of septic shock and intensive care unit admission were significantly reduced compared to controls ( P = 0.011 and 0.016, respectively). No infection-associated mortality was found after the implementation of the guidelines. Conclusions: Using the clinical practice guidelines for pediatric oncology patients with fever was found to reduce the adverse outcomes and improve survival. [ABSTRACT FROM AUTHOR]

Watcharananan SP, Kiertiburanakul S, Piyatuctsanawong W, Anurathapan U, Sungkanuparph S, Pakakasama S, Chantratita W, Hongeng S. Cytomegalovirus, adenovirus, and polyomavirus co-infection among pediatric recipients of allogeneic stem cell transplantation: Characteristics and outcome. Pediatr Transplantation 2010: 14:675–681. © 2010 John Wiley & Sons A/S. ADV and PMV infection have increasingly been documented as significant complications following allo-HSCT. Despite increasing recognition, characteristics and outcome of CMV, ADV, and PMV viral co-infection remain obscured. In this study, a retrospective quantitative PCR analysis of ADV, PMV (BKV and JCV) was performed from pediatric patients’ stored blood samples previously tested for CMV viremia after allo-HSCT. Clinical and virological characteristics and outcome among patients with and without viral co-infection were analyzed and compared. From 2001 to 2006, 219 blood samples from 69 patients were studied. Viral DNA was present in 119 samples (52.9%).The proportion of viremia was highest for BKV (30.6%), followed by CMV (20.9%), ADV (9.1%), and JCV (0.5%). Viral co-infection occurred in 17 patients (24.6%), with CMV/BKV as the most common type (11.6%), followed by CMV/ADV (4.3%) and ADV/BKV (2.9%). From multivariate analysis, factors associated with viral co-infection were acute GVHD (OR 4.57; 95% CI 1.9–10.96, p = 0.001), level of blood CMV viral load (OR 1.53; 95% CI 1.24–1.89, p < 0.001), and level of blood ADV viral load (OR 1.56; 95% CI 1.05–2.32, p = 0.027). Higher probability of developing viral disease was strongly associated with more types of virus detected in blood (p < 0.001). Significant difference in the causes of death was observed among patients with and without viral co-infection (p = 0.014). Infection (87.5%) was the major cause of death of patients with viral co-infection, whereas relapse of hematologic disease (70%) was the major cause of death of patients with mono-viral infection. Viral co-infection is a common and significant infectious complication in pediatric recipients of allo-HSCT. Blood monitoring of CMV, ADV, and BKV is suggested among pediatric patients who develop GvHD or who have rising of CMV or ADV viremia following allo-HSCT. [ABSTRACT FROM AUTHOR]

Aim: Polymorphisms of the UGT1A1 gene, SLCO1B1 gene and GST gene have been associated with significant hyperbilirubinemia. We would like to determine whether the variation of UGT1A1 gene, SLCO1B1 gene and GST gene may play a significant role in neonatal hyperbilirubinemia in Thai infants. Methods: Ninety-one study subjects (hyperbilirubinemic group) and 86 control subjects were studied. Results: The cause of neonatal hyperbilirubinemia could not be identified in 64 infants (70.3%), ABO blood group incompatibility in 14.3% and Glucose-6-phosphate dehydrogenase (G-6-PD) deficiency in 8.8%. In the hyperbilirubinemic group, 23 of 91 (25.3%) infants demonstrated variant of UGT1A1 at nucleotides (nt) 211 as compared to 6 of 86 (7%) in the control group (p = 0.001). There were no significant differences between groups in the variants UGT1A1 at nt 686, SLCO1B1 gene at nt 388, 463 and the GST gene. Male infants with G-6-PD deficiency were associated with hyperbilirubinemia (21.2% vs. 4.8% in the control group) with an odds ratio (OR) of 5.37 (p =0.02). The relationship between G-6-PD and variant in UGT1A1 gene at nt 211 could not be determined. Conclusion: Thai infants with variant in the UGT1A1 at nt 211 or with G-6-PD deficiency are at higher risk for developing neonatal hyperbilirubinemia. [ABSTRACT FROM AUTHOR]

EBV-PTLDs affect as high as 20% of SCT recipients especially those with T-cell depleted grafts while high mortality rates were also noted. Adoptive allogeneic and autologous CTLs have a therapeutic potential in this setting. However, the process of expansion of these cells is tedious and time consuming in both allogeneic and autologous CTL generation. For the allogeneic SCT, another major obstacle is unavailability of donors especially in an unrelated SCT setting. The aim of the present study was therefore to investigate the efficacy of autologous CIK cells (CD3+ CD56+) against autologous EBV-LCLs from post-SCT pediatric patients. We could demonstrate that CIK cells can be generated within two wk and did show the significant cytotoxicity against autologous EBV-LCLs. CIK cells may provide a potent tool for use in post-transplantation adoptive immunotherapy. [ABSTRACT FROM AUTHOR]

Abstract: Osteosarcoma with distant metastases at late stage has posed a challenge for novel therapeutic modalities. The application of cytokine-induced killer (CIK) cells to osteosarcoma constitutes a promising strategy. This approach had been studied in multiple myeloma and breast cancers, where CIK cells exhibited specific cytotoxicity toward malignant cells while sparing wild-type tissues. However, the consistency of CIK cell-induced anti-tumor cytotoxicity has not been thoroughly examined. We investigated whether autologous CIK cells could effectively induce cytolysis of cultured osteosarcoma cells. In addition to the observed CIK cell-induced osteosarcoma cytolysis, the pre-incubation of CIK cells with autologous dendritic cells pulsed with tumor’s total RNA further enhanced the tumor cytolysis to greater than 6-fold. The anti-tumor cytolysis was optimized in complete autologous setting, and was attenuated with allogeneic components. The advantage of the co-culture with RNA-pulsed DC was lost when high CIK cell density was employed for anti-tumor cytotoxic assay, but was maintained in purified CD3+CD56+ cells isolated from the CIK cells. This finding implied that CIK cells at limited cell density could induce effective osteosarcoma cytolysis with an aid from tumor antigen presentation on dendritic cell surface. [Copyright &y& Elsevier]

Choeyprasert W, Pakakasama S, Anurathapan U, Songdej D, Sirachainun N, Sirireung S, Panthangkool W, Hongeng S. The second mini-transplant for unstable mixed chimerism within the first 100 days after hematopoietic stem cell transplant in severe thalassemia. Pediatr Transplantation 2011. © 2011 John Wiley & Sons A/S. Abstract: Allogeneic HSCT is the only curative treatment for severe thalassemia disease. MC occurs in one-third of these patients within the first two months after HSCT; this is a major risk factor of graft rejection, especially when RHCs are more than 25%. There is still no consensus for the management of MC, especially in the early phase of HSCT. The DLI has also been described in the treatment of MC following HSCT for hemoglobinopathies, but its success is still not guaranteed. The second HSCT has been an approach used in an attempt to cure patients who reject their graft. Concern about toxicity of conditioning regimen, the second HSCT is usually delayed for at least a year after the first HSCT. We would like to demonstrate the successful use of the second mini-allogeneic HSCT in hemoglobin E/β-thalassemia with evidence of unstable MC in the first 100 days after allogeneic HSCT to prevent further graft loss after allogeneic HSCT. [ABSTRACT FROM AUTHOR]

Introduction Methods Results Conclusion Immune reconstitution (IR) kinetics of paediatric patients underwent haploidentical haematopoietic stem cell transplantation (HSCT) with post‐transplant cyclophosphamide (PTCy) have not been extensively studied. We compared IR patterns of children receiving HSCT from haploidentical (n = 92) and HLA‐matched donors (n = 36), and analysed risk factors for viral infection in these patients.We prospectively measured lymphocyte subset numbers before HSCT and at 1, 3, 6 and 12 months after HSCT. Blood cytomegalovirus (CMV), Epstein–Barr virus, adenovirus, BK virus (BKV) and urine adenovirus and BKV viral loads were measured at designated time points.The median numbers of total T and T helper cells at 1 month were significantly lower in the haploidentical group compared with the HLA‐matched group. Haploidentical HSCT recipients had significantly lower median numbers of several T cell subsets and B cells for 1 year after HSCT. The median NK cell count of the haploidentical group was lower at 1 month. BKV haemorrhagic cystitis, blood CMV and urine adenovirus reactivation were more frequently found in the haploidentical group. Post‐haploidentical HSCT patients receiving anti‐T lymphocyte globulin (ATG) had significantly lower median numbers of total T cells (at 1 month) and T helper cells (at 6 and 12 months) and higher rate of blood BKV reactivation compared with those without ATG.Paediatric patients who undergo haploidentical HSCT with PTCy are likely to have delayed IR and an increased risk of viral reactivation/infection compared with HLA‐matched HSCT. The addition of ATG to PTCy delayed T cell recovery and increased risk of BKV reactivation. [ABSTRACT FROM AUTHOR]