Prevalence of massively diluted bone marrow cell samples aspirated from patients with myelodysplastic syndromes (MDS) or suspected of MDS: A retrospective analysis of nationwide samples in Japan.

Bone marrow (BM) examination is a key element in the diagnosis and prognostic grading of myelodysplastic syndromes (MDSs), and obtaining adequate BM cell samples is critical for accurate test results. Massive haemodilution of aspirated BM samples is a well-known problem; however, its incidence in patients with MDS has not been well studied. We report the first study to examine the incidence of massive haemodilution in nationwide BM samples aspirated from patients diagnosed with or suspected of MDS in Japan. Among 283 cases available for analysis, BM smears from 92 cases (32.5%) were hypospicular (massively haemodiluted) and, particularly, no BM particles were observed in 52 cases (18.4%). Regarding hypospicular cases, we examined how the doctors in charge interpreted the BM smears of their patients. In only 19 of 92 cases (20.7%), doctors realised that the BM smears were haemodiluted. Furthermore, the BM biopsy, which can help diagnose hypospicular cases, was oftentimes not performed when the haemodilution was overlooked by doctors (not performed in 50 of 73 such cases). These real-world data highlight that not only researchers who are working to improve diagnostic tests but also clinicians who perform and use diagnostic tests must realise this common and potentially critical problem.

[DEVELOPMENT AND VALIDATION OF QUALITY OF LIFE QUESTIONNAIRE FOR PARENT BURDEN OF CHILDREN WITH FOOD ALLERGY].

OBJECTIVE: Quality of life (QOL) questionnaires for parents of children with food allergies have been developed in the United States and Europe. However, no original Japanese QOL questionnaire has been developed till date. We aimed to develop an original questionnaire to evaluate the QOL in parents of children with food allergies in Japan. METHODS: We collected QOL-related questions from parents of children with food allergies aged 0-15 years, and created a primary questionnaire. Responses to the primary questionnaire were obtained from the parents again, and question items were reduced using factor analysis to create a secondary questionnaire comprising eight items. In addition to the secondary questionnaire, responses to the Food Allergy QOL Questionnaire-Parent Form (FAQLQ-PF) Japanese version, Parent reported Health-Related QOL in children and adolescents (KINDL) and Health-related QOL (SF-8) were obtained from parents to assess the validity of the secondary questionnaire. RESULTS: A total of 407 parents completed all questionnaires. The secondary questionnaire scores were positively correlated with those of FAQLQ-PF and weakly negatively correlated with the KINDL and SF-8 mental component summary scores. Parents of children with food allergies with ≥3 culprit foods or severe reactions to daily foods, a history of anaphylaxis, and those carrying adrenaline autoinjectors scored higher and had lower QOL. CONCLUSION: The developed original questionnaire is a valid QOL questionnaire for Parents of children with food allergies.

Safety of Oral Food Challenge for Individuals with Low Egg White and Ovomucoid-Specific IgE Antibodies.

INTRODUCTION: During an oral food challenge (OFC), there is a risk of adverse reactions, including anaphylaxis. Therefore, the physician should carefully conduct the OFC. This study aimed to evaluate the OFC results in individuals with low levels of egg white (EW)- and ovomucoid (OVM)-specific immunoglobulin E (sIgE) and the safety of a hen's egg (HE) OFC in these individuals. METHODS: A total of 2,058 individuals with low EW- or OVM-sIgE underwent HE-OFC at two institutions in Kumamoto prefecture, located in the western area of Japan, between January 1, 2017, and December 31, 2021, within 1 year of recorded sIgE measurements. The ImmunoCAP systems were used to measure sIgEs. The HE-OFC test was performed according to the 2017 Food Allergy Guidelines in an open and unblinded method. RESULTS: Five hundred and one individuals (24.3%) had low EW-sIgE levels (class 2 or lower), and 926 (45.0%) had low OVM-sIgE levels (class 2 or lower). Individuals with low EW-sIgE had lower total IgE and OVM-sIgE than did those with high EW-sIgE (greater than class 2). Those with low OVM-sIgE had lower total IgE and EW-sIgE than did those with high OVM-sIgE (greater than class 2). Among the individuals with low EW-sIgE, 86.4% (433/501 cases) passed the OFC without symptoms. Among the individuals with low OVF-sIgE, 82.6% (765/926 cases) passed the OFC without symptoms. CONCLUSION: More than 80% of individuals with suspected IgE-dependent HE allergy and low levels of EW- or OVM-specific IgE were able to consume at least a small amount of HE. As the OFC results are independent of the loading dose in cases with low EW- or OVM-sIgE, a medium-dose HE-OFC may be performed safely in individuals with no history of anaphylaxis.

[JAPANESE TRANSLATION AND LINGUISTIC VALIDATION OF THE RECAP OF ATOPIC ECZEMA (RECAP)].

BACKGROUND: The Recap of atopic eczema (RECAP), a new core outcome of the atopic dermatitis trial, was translated into Japanese and linguistically validated. METHODS: Translation into Japanese was accomplished according to the ISPOR (International Society for Pharmacoeconomics and Outcome Research) guidelines and the basic guidelines for scale translation. The translation process included two forward translations, reconciliation with native English speakers, third-party back translation, cognitive debriefing, review and harmonization by the original authors. Twenty-seven atopic dermatitis and pediatric specialists from 21 centers in Japan participated in the translation process. Cognitive debriefing was conducted through face-to-face interviews using a think-aloud method with the interview guide including questions about comprehensibility, relevance, comprehensiveness, recall period and suggested improvements, based on the COSMIN methodology. RESULTS: No linguistic or cultural problems were encountered in the translation into Japanese. Cognitive debriefings were conducted with 10 adult patients and 10 parents of pediatric patients. Some minor modifications were made following discussion and approval by the research team and the original authors. The Japanese version of RECAP was considered to be understandable, comprehensive and relevant for adult patients and families of pediatric patients. CONCLUSION: The Japanese version of the RECAP, which has been validated as linguistically equivalent to the original version, is now available. Further evaluation of the measurement properties is needed in the future.

Myeloblasts transition to megakaryoblastic immunophenotypes over time in some patients with myelodysplastic syndromes.

OBJECTIVES: In myelodysplastic syndromes (MDS), neoplastic myeloblast (CD34+CD13+CD33+ cells) numbers often increase over time, leading to secondary acute myeloid leukemia (AML). In recent studies, blasts in some MDS patients have been found to express a megakaryocyte-lineage molecule, CD41, and such patients show extremely poor prognosis. This is the first study to evaluate whether myeloblasts transition to CD41+ blasts over time and to investigate the detailed immunophenotypic features of CD41+ blasts in MDS. METHODS: We performed a retrospective cohort study, in which time-dependent changes in blast immunophenotypes were analyzed using multidimensional flow cytometry (MDF) in 74 patients with MDS and AML (which progressed from MDS). RESULTS: CD41+ blasts (at least 20% of CD34+ blasts expressing CD41) were detected in 12 patients. In five of these 12 patients, blasts were CD41+ from the first MDF analysis. In the other seven patients, myeloblasts (CD34+CD33+CD41- cells) transitioned to megakaryoblasts (CD34+CD41+ cells) over time, which was often accompanied by disease progression (including leukemic transformation). These CD41+ patients were more frequently observed among patients with monosomal and complex karyotypes. CD41+ blasts were negative for the erythroid antigen, CD235a, and positive for CD33 in all cases, but CD33 expression levels were lower in three cases when compared with CD34+CD41- blasts. Among the five CD41+ patients who underwent extensive immunophenotyping, CD41+ blasts all expressed CD61, but two cases had reduced CD42b expression, three had reduced/absent CD13 expression, and three also expressed CD7. CONCLUSIONS: Myeloblasts become megakaryoblastic over time in some MDS patients, and examining the megakaryocyte lineage (not only as a diagnostic work-up but also as follow-up) is needed to detect CD41+ MDS. The immunophenotypic features revealed in this study may have diagnostic relevance for CD41+ MDS patients.

Clinical, immunophenotypic, and cytogenetic characteristics of high-grade myelodysplastic syndromes with CD41-positive progenitor cells.

BACKGROUND: Patients with myelodysplastic syndromes (MDS) with progenitors expressing CD41 (CD41+ MDS) showed a poor prognosis in a previous study but their detailed characteristics remain unclear. METHODS: One hundred thirty-seven subjects at our institution were diagnosed with excess blasts (EB)-1, EB-2, and acute myeloid leukemia with a low blast count (20%-30%). The immunophenotypes of progenitor cells in their bone marrow (BM) were determined by CD45-gating flow cytometry. A false-positive reaction to CD41 was eliminated by examining the flow cytometry data of lymphocytes and monocytes in addition to progenitors and by examining CD42b in histological sections. The characteristics were compared between CD41+ and CD41- MDS patients. RESULTS: Forty-three patients (31%) were CD41+. Additionally, 91% of the CD41+ MDS patients were very high-risk defined by the Revised International Prognostic Score System, which was higher than in patients with CD41- MDS (p = 0.015). Approximately 60% of the CD41+ MDS patients had a monosomal karyotype and very poor cytogenetics, which was higher than in CD41- MDS patients (p < 0.001). Normal cytogenetics was less common in CD41+ patients (p = 0.0016). Blasts with bleb formation were more abundant in CD41+ MDS patients (p = 0.026). All CD41+ MDS patients were positive for CD13 and were mostly positive for CD33. The frequency of aberrant expression of other antigens on progenitors was similar between CD41+ and CD41- MDS patients. CONCLUSIONS: We determined clinical, immunophenotypic, and cytogenetic characteristics of CD41+ MDS patients. Further studies are needed to improve the survival of these patients.

Food-Dependent Exercise-Induced Anaphylaxis Caused by Carrots: A Case Report.

Background: Most cases of food-dependent exercise-induced anaphylaxis (FDEIA) are caused by eating wheat or crustaceans. However, fruits or vegetables may rarely act as allergens for FDEIA. We report a rare case of FDEIA caused by eating carrots. Case Presentation: An 8-year-old boy developed an anaphylactic reaction while playing, after eating lunch that included cooked carrots. Serum carrot-specific immunoglobulin E level was 0.19 UA/mL. The prick-by-prick test for raw carrots was positive (wheal diameter: 4 mm). The patient developed urticaria after exercise provocation tests following ingestion of raw carrots. Carrot proteins were analyzed by 2-dimensional Western blotting to identify the causative allergens. Nine proteins were identified as candidate antigens at 21-66 kDa. Conclusions: Our patient presented with FDEIA symptoms after ingesting both raw and cooked carrots. Both raw and cooked carrots contain 9 proteins that may induce FDEIA.

Oral Immunotherapy for Children with Cow's Milk Allergy.

Cow's milk allergy (CMA) is one of the most common IgE-dependent food allergies in children. Some children develop severe and persistent CMA, with near-fatal reactions after exposure to trace amounts of cow's milk (CM). Because milk and dairy products are included in various processed food products, it is difficult to completely remove milk, which negatively affects the quality of life of children with CMA. Oral immunotherapy (OIT) can alleviate food allergen-induced anaphylaxis under continuous ingestion of a little of the causative food. Children with severe CMA may benefit from OIT, but the treatment requires a long time and poses a risk of anaphylaxis. Moreover, in recent years, new therapies, including omalizumab, sublingual immunotherapy, and epicutaneous immunotherapy, have played the role of optional OIT. In this review, we present the current methods of and other attempts at OIT, and discuss OIT for safely treating CMA.

Hyperglycemic hyperosmolar state in an adolescent with type 1 diabetes mellitus.

Hyperglycemic hyperosmolar state (HHS) and diabetic ketoacidosis (DKA) are the most severe acute complications of diabetes mellitus (DM). HHS is characterized by severe hyperglycemia and hyperosmolality without significant ketosis and acidosis. A 14-year-old Japanese boy presented at the emergency room with lethargy, polyuria and polydipsia. He belonged to a baseball club team and habitually drank sugar-rich beverages daily. Three weeks earlier, he suffered from lassitude and developed polyuria and polydipsia 1 week later. He had been drinking more sugar-rich isotonic sports drinks (approximately 1000-1500 mL/day) than usual (approximately 500 mL/day). He presented with HHS (hyperglycemia (1010 mg/dL, HbA1c 12.3%) and mild hyperosmolality (313 mOsm/kg)) without acidosis (pH 7.360), severe ketosis (589 µmol/L) and ketonuria. He presented HHS in type 1 diabetes mellitus (T1DM) with elevated glutamate decarboxylase antibody and islet antigen 2 antibody. Consuming beverages with high sugar concentrations caused hyperglycemia and further exacerbates thirst, resulting in further beverage consumption. Although he recovered from HHS following intensive transfusion and insulin treatment, he was significantly sensitive to insulin therapy. Even the appropriate amount of insulin may result in dramatically decreasing blood sugar levels in patients with T1DM. We should therefore suspect T1DM in patients with HHS but not those with obesity. Moreover, age, clinical history and body type are helpful for identifying T1DM and HHS. Specifically, drinking an excess of beverages rich in sugars represents a risk of HHS in juvenile/adolescent T1DM patients. Learning points: Hyperglycemic hyperosmolar state (HHS) is characterized by severe hyperglycemia and hyperosmolality without significant ketosis and acidosis. The discrimination between HHS of type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM) in initial presentation is difficult. Pediatrician should suspect T1DM in patients with HHS but not obesity. Age, clinical history and body type are helpful for identifying T1DM and HHS. Children with T1DM are very sensitive to insulin treatment, and even appropriate amount of insulin may result in dramatically decreasing blood sugar levels.

Administration of the adrenaline auto-injector at the nursery/kindergarten/school in Western Japan.

BACKGROUND: In view of the increasing prevalence of food allergies, there has been an associated increase in frequency of situations requiring an emergency response for anaphylaxis at the home, childcare facilities and educational institutions. OBJECTIVE: To clarify the situation of adrenaline auto-injector administration in nursery/kindergarten/school, we carried out a questionnaire survey on pediatric physicians in Western Japan. METHODS: In 2015, self-reported questionnaires were mailed to 421 physicians who are members of the West Japan Research Society Pediatric Clinical Allergy and Shikoku Research Society Pediatric Clinical Allergy. RESULTS: The response rate was 44% (185 physicians) where 160 physicians had a prescription registration for the adrenaline auto-injector. In the past year, 1,330 patients were prescribed the adrenaline auto-injector where 83 patients (6% of the prescribed patients) actually administered the adrenaline auto-injector, of which 14 patients (17% of the administered patients) self-administered the adrenaline auto-injector. "Guardians" at the nursery/kindergarten and elementary school were found to have administered the adrenaline auto-injector the most. Among 117 adrenaline auto-injector prescription-registered physicians, 79% had experienced nonadministration of adrenaline auto-injector at nursery/kindergarten/school when anaphylaxis has occurred. The most frequent reason cited for not administering the adrenaline auto-injector was "hesitation about the timing of administration." CONCLUSION: If the adrenaline auto-injector was administered after the guardian arrived at the nursery/kindergarten/school, it may lead to delayed treatment of anaphylaxis in which symptoms develop in minutes. Education and cooperation among physicians and nursery/kindergarten/school staff will reduce the number of children suffering unfortunate outcomes due to anaphylaxis.

The skin prick test is not useful in the diagnosis of the immediate type food allergy tolerance acquisition.

BACKGROUND: Some studies have been reported about positioning of SPT in the diagnosis of food allergy. On the other hand, it is not yet clear about the positioning of SPT in the diagnosis of tolerance acquisition of the immediate type food allergy. METHODS: The retrospective study had been conducted for 236 egg allergic children (51.3 months in mean), 127 milk allergic children (53.4 months), and 96 wheat allergic children (42.6 months). The retrospective analysis of serum nonspecific and antigen-specific IgE levels, SPT, and OFC had been conducted for each allergic patient. All OFC had been conducted to verify the acquisition of tolerance against eliminated food. RESULTS: The OFC was positive in 61 (25.8%) hen's egg allergies, 34 (26.8%) milk allergies and 33 (34.4%) wheat allergies. The greatest AUC for each allergen is as following; hen's egg for egg white specific IgE at 0.745, milk/histamine (wheal) index at 0.718, and wheat for wheal size at 0.597. For the predictive decision points, the highest accuracy rate was at 25.8% for an egg white wheal of 9.5mm, at 26.8% for a milk wheal of 9.5mm, and at 34.4% for a wheal of 6.5mm. CONCLUSIONS: As a result of this analysis, the diagnostic accuracy of SPT had not been satisfactory to judge the acquisition of tolerance in allergic children for eggs, milk and wheat. Therefore, this is not a strong evidence to testify the tolerance of the immediate type food allergy.

[Lupus anticoagulant-hypoprothrombinemia syndrome with a false-positive test for coagulation factor inhibitors].

We report a case of a 1-year-old boy diagnosed with lupus anticoagulant-hypoprothrombinemia syndrome (LA-HPS), which is a rare disorder. His initial presentation of sinusitis was accompanied by hemorrhagic episodes including ecchymoses and epistaxis 6 months after antibiotic therapy. Laboratory results revealed prolonged prothrombin time (PT) and activated partial thromboplastin time (APTT) that did not correct with mixing studies. Factors II, VIII, IX, X, XI, and XII activities were 20%, 44%, 42.5%, 59%, 4%, and 10%, respectively. The Bethesda inhibitor assay showed inhibitors against multiple coagulation factor. APTT, mixing studies, diluted Russell's viper venom time, and the Bethesda inhibitor assay detected LA. LA-HPS with a suspected false-positive test for coagulation factor inhibitors was diagnosed. Bleeding stopped and results of coagulation studies returned to normal without therapy 2 months after onset of the disease.

Utility of the peripheral blood basophil histamine release test in the diagnosis of hen's egg, cow's milk, and wheat allergy in children.

BACKGROUND: The diagnosis of food allergy (FA) is made by oral food challenge tests (OFCs) that occasionally produce serious symptoms in patients; therefore, whether to perform OFCs should be carefully considered. The utility of the histamine release test (HRT) in the diagnosis of childhood FA has not been fully examined. METHODS: Sixty-four subjects with suspected hen's egg allergy, cow's milk allergy (CMA), and wheat allergy (WA) were enrolled. The diagnosis of FA was made based on the outcomes of OFCs or a convincing history of symptoms after food ingestion within 6 months before or after sample collection. HRT was performed using an HRT Shionogi kit. The threshold of histamine release (HRT threshold), which was defined as the minimum concentration of food antigen to induce a 10% net histamine release, was analyzed in association with FA diagnosis. RESULTS: Receiver operating characteristic analysis showed that the HRT threshold was useful in the diagnosis of heated egg allergy (HEA), raw egg allergy (REA), CMA, and WA. We were able to determine the cutoff value for the HRT threshold in relation to outcomes of OFCs. The cutoff value was 6 ng/ml of egg white antigen in HEA and REA (p < 0.01), 40 ng/ml of milk antigen in CMA (p < 0.01), and 500 ng/ml of wheat antigen in WA (p < 0.05). The efficiency was 70.3% for HEA, 78.0% for REA, 77.6% for CMA, and 70.7% for WA. CONCLUSIONS: We conclude that the HRT threshold measurement for egg white, milk, and wheat antigen is related to outcomes of OFCs and is useful in determining when OFCs should be performed.

[Usefulness of skin prick test using bifurcated needle for the diagnosis of food allergy among infantile atopic dermatitis--second report. In the case of cow's milk allergy].

OBJECTIVE: We investigated the usefulness of skin prick test (SPT) using bifurcated needle (BN) for the diagnosis of cow's milk allergy (CMA) in early infancy. SUBJECTS AND METHODS: Among atopic dermatitis infants with suspicion of food allergy who visited Sagamihara National hospital from January 2001 to April in 2005, 168 cases had received both SPT and IgE ImmunoCAP of CM (CM-IgE) at first visit. We analyzed results of SPT and CM-IgE, focused on infants with negative CM-IgE to examine the usefulness of SPT, and checked positive conversion of CM-IgE with aging. RESULTS: Among 124 infants (73.8%) with negative CM-IgE, 34 infants (27.4%) showed positive SPT results. Forty two infants (33.6%) among the 124 cases were diagnosed as CMA by combination of elimination and provocation test, and 21 infants (50.0%) had positive SPT. In the follow up study of 39 negative CM-IgE cases with CMA, 21 CM-IgE (53.8%) turned positive later infantile period (mean CM-IgE: 4.2+/-4.8 Ua/ml at 9.3+/-5.3 months old). CONCLUSIONS: SPT using BN seemed to be more useful than CM-IgE for the diagnosis of CMA in early infancy with AD. But, the sensitivity is lower than in the egg case.

[Usefulness of skin prick test using bifurcated needle for the diagnosis of food allergy in infantile atopic dermatitis--1st report. Case of egg allergy].

OBJECTIVE: We investigated the usefulness of skin prick test (SPT) for the diagnosis of egg white (EW) allergy in infants with atopic dermatitis who showed negative to EW CAPRAST, and followed up the EW-CAPRAST in this study. SUBJECTS AND METHODS: Data of negative SPT using Bifurcated needle (BN) were analyzed from the data of 202 atopic dermatitis infants, who had received SPT from January in 2001 to April in 2005. From the negative SPT value (average and standard deviation) positive SPT value was obtained. Among 202 cases, 89 suspected-egg allergy infants with negative IgE CAPRAST against EW at the time of first visit were recruited to examine the usefulness of SPT. Positive conversion of EW-CAPRAST was checked in 78 cases (65: egg allergy+, 13: egg allergy(-)) who had been followed up in our outpatient clinic. RESULTS: Range of negative SPT control value (mean+2SD) using BF among infants could be set as less than 2 mm for wheal and/or 5 mm for erythema. Among 89 suspected-egg allergy infants with negative EW-CAPRAST, 72 infants (80.9%) were diagnosed as egg allergy by the combination of elimination and provocation test, interestingly 39 infants (54.2%) showed positive SPT results. In the follow up study of 78 negative EW-CAPRAST cases, 47 EW-CAPRAST out of 65 egg-allergy cases turned positive later infantile period (mean EW-CAPRAST: 9.6+/-16.7 Ua/ml at 9.9+/-5.6 months old). EW-CAPRAST of 7 cases in 13 non-egg allergies also turned positive in the follow up, however EW-CAPRAST titer was relatively lower compared to that of egg allergies (1.1+/-1.5 Ua/ml at 13.3+/-2.6 months old). CONCLUSIONS: We experienced fairly number of atopic infants with negative EW-CAPRAST at the first outpatient visit, who were later diagnosed as egg allergy. In about half of these cases, SPT egg-allergy infants, three quarter of EW-CAPRAST turned positive around 10 months old. EW-CAPRAST of atopic infants without egg allergy also turned transiently and slightly positive. In the conclusions, SPT seemed to be more useful than EW-CAPRAST for the diagnosis of egg allergy in early infantile period, however provocation test should be required for the definitive diagnosis in suspected-egg allergy infants without any proof of egg-sensitization.

[Prolonged type of food allergy].

OBJECT: Prevalence of food allergy is the most frequent during infancy, and it is gradually decreasing with age. We can not distinguish patients developing tolerance from those with persistent food allergy, therefore we are unable to advice patients accelerating the development of tolerance. To clarify the factors developing tolerance or intolerance, we conducted the following study. METHOD: Patients who were diagnosed as food allergy to hen's egg, or cow's milk or wheat in infancy by the definitive history of positive food allergic reactions or food provocation tests were recruited to this study. Patients were divided into two groups; one (prolonged group, n=27) is those who needed to eliminate some of the main offending foods even at the age of 6 years old and the other (tolerized group, n=37) is those who had released all main offending foods by the age of 6 years old. RESULT: The factors which distinguished the prolonged group from the tolerized group were the positive clinical history of the atopic dermatitis and its prolonged clinical course, past history of anaphylactic shock, and maximum number of offending foods in the past clinical history. The specific IgE against main antigens such as egg white, cow's milk and wheat in the tolerized group was lower compared to that in the prolonged group, whereas no significant difference was found in non specific IgE value, peripheral eosinophil counts, and specific IgE against other antigens. CONCLUSION: We could find the clinical factors discriminating food allergy patients against three major food allergen among children developing tolerance by the age of 6 years old form those without tolerance.

Antiproliferative activity of root extract from gentian plant (Gentiana triflora) on cultured and implanted tumor cells.

We describe a novel pharmacological activity of the gentian root, an ingredient of Chinese medicines. Root extract from Gentiana triflora triggered cell death of human Daudi cells in culture. In addition, daily administration of the extract to mice inhibited growth of implanted solid tumors. Extract treatment of cultured cells resulted in the appearance of shranken, fragmented, or condensed cell and nuclear morphologies, and in chromosomal DNA degradation. But, the extract-treated cells did not show DNA fragmentation, which exhibits a nucleosome ladder, suggesting that extract-triggered cell death is not mediated through a typical apoptotic pathway.