Clinical validation of an immunochromatographic SARS-Cov-2 IgM/IgG antibody assay with Japanese cohort.

BACKGROUND: The coronavirus disease 2019 (COVID-19) has become a major health threat. To overcome COVID-19, appropriate diagnosis methods are urgently needed. The aim of this study was to clinically evaluate the colloidal gold immunochromatography assay for SARS-Cov-2 IgM/IgG antibody (Ab). METHODS: Patients confirmed COVID-19 (n = 51) were recruited prospectively from the Musashino Red Cross hospital and Tokyo Medical and Dental University Medical Hospital, between March and May 2020. And the analytical specificity was assessed with serum samples of patients without COVID-19 (n = 100) collected between August to September 2019 before SARS-CoV-2 was first reported in China. RESULTS: Among COVID-19 patients, a total of 87 serum samples were tested for SARS-Cov-2 IgM/IgG Ab assay. IgM was detected 71.0 %, 86.9 %, and 83.3 % at day8-14, 15-28, >29 after symptom onset and IgG was detected in 81.6 %, 87.0 %, and 94.4 %, respectively. The sensitivity of IgM and IgG Ab after day8 assay was significantly higher than before day7, respectively (p=0.0016, 0.0003). There were no positive results in 100 serum samples from patients without COVID-19. CONCLUSION: The SARS-Cov-2 IgM/IgG Ab assay had 79.7% / 86.1% sensitivity (the 8 days after from onset) and 100% specificity in this population.

[Sustained deep molecular response over eight years after discontinuation of imatinib for chronic myeloid leukemia].

A 72-year-old female was diagnosed with chronic phase chronic myeloid leukemia (CML) in 2001. After a short course of treatment with hydroxycarbamide, imatinib (IM) 400 mg was started. A major molecular response was presumably acquired 10 months later. IM was discontinued after treatment for 47 months in November 2005. At the same time, BCR-ABL transcript was undetectable by nested RT-PCR assay, which was equivalent to <0.00138% BCR-ABL according to the international scale. The patient is still under observation with no additional therapy, and BCR-ABL has remained negative for 102 months, to date. Furthermore, interferon was never used in this patient. IM has dramatically improved the prognosis of CML. Since no cure has yet been established, patients are recommended to continue treatment even after achieving deep molecular responses. There are several ongoing clinical trials challenging the discontinuation of IM, but long-term observation is still lacking. The sustained deep molecular response, exceeding eight years, experienced in this patient is potentially encouraging.

Nationwide Descriptive Epidemiological Study of Patients with COVID-19 Evacuated from Wuhan, China to Japan from January to February, 2020.

We investigated the epidemiological findings regarding the route of coronavirus disease 2019 (COVID-19) and infection prevention and control (IPC) measures among returnees in the emergency evacuation from Wuhan, China to Japan during the COVID-19 outbreak in 2020. A total of 12 of the 14 returnees (median age [range]: 49.5 years [29-65 years]; 9 men [75%]) had confirmed COVID-19. The proportion of returnees with COVID-19 was 12/566 (2.1%) in Flights 1-3 and 2/263 (0.8%) in Flights 4 and 5. Six patients were asymptomatic on admission, while 3 patients developed symptoms thereafter. None of the participants reported a specific history of contact with animals, going to seafood markets, or visiting medical facilities. Two patients were in contact with an individual who was confirmed or suspected of having COVID-19. Most patients resided in hotels in the center of Wuhan City, taking taxis and trains for commute. Patients relatively adhered to IPC measures such as wearing a mask and hand hygiene. However, emphasis on IPC measures such as universal masking and more rigorous avoidance of exposure risk might have been necessary to prevent infection. In addition, forced social distancing due to lockdown might have contributed to the lower infection rates in Flights 4 and 5, compared to Flights 1-3.

A severe case of thrombocytopenia, anasarca, fever, renal insufficiency or reticulin fibrosis, and organomegaly syndrome with myocardial and skeletal muscle calcification despite hypocalcemia: a case report.

BACKGROUND: TAFRO (thrombocytopenia, anasarca, fever, renal insufficiency or reticulin fibrosis, and organomegaly) syndrome is a recently recognized disease with a variety of presentations of variable severity. In acute settings, this disease also involves organ dysfunction because of the associated systemic inflammation. However, cases of TAFRO syndrome with myocardial and/or skeletal muscle calcification have never been reported. CASE PRESENTATION: A 24-year-old healthy young Asian man was admitted with intermittent epigastric pain and fever for 2 weeks. Computed tomography revealed pleural effusion, ascites and systemic lymphadenopathy. Laboratory tests showed thrombocytopenia, elevated C-reactive protein, hypoalbuminemia, anemia and renal dysfunction. Based on these findings and bone marrow biopsy, we diagnosed his disease as TAFRO syndrome and commenced hemodialysis for the renal dysfunction. However, he developed refractory hypocalcemia with unstable vital signs, for which we administered calcium gluconate hydrate. Thereafter, myocardial and skeletal muscle calcification was revealed radiologically, with the myocardial calcification causing sick sinus syndrome. He was treated with tocilizumab and finally discharged in an ambulatory condition after prolonged hospitalization, with residual calcific lesions. CONCLUSION: This is the first report of a patient with TAFRO syndrome and the complication of organ calcification. The etiology of calcification in this case is not clear. Systemic inflammation with possible hypercytokinemia might have been involved in the unexpected complication of systemic calcification. It is important to carefully handle the general management of TAFRO syndrome because of the possibility of various complications.

Investigation of Anti-SARS-CoV-2 IgG and IgM Antibodies in the Patients with COVID-19 by Three Different ELISA Test Kits.

We examined anti-SARS-CoV-2 IgG and IgM antibodies in 45 serum samples from 26 patients with COVID-19, who were admitted in our hospital by using three different ELISA kits. All patients had pneumonia at admission, and 7 patients required mechanical ventilator support and grouped in severe case. Anti-SARS-CoV-2 IgG and IgM antibodies turned to be partially positive between the 6th and 10th days, more than 84% positive between the 11th and 15th days, and 100% after the 16th day. One ELISA kit revealed poorer sensitivity for anti-SARS-CoV-2 IgM antibody. Negative conversion of IgM antibody was not observed in the 30th day in our cohort. All three ELISA kits showed no false positive reaction for negative serum samples. Between severe and moderate cases, there was no significant difference in the trends of anti-SARS-CoV-2 IgG and IgM antibody.

Liver injury with COVID-19 based on gastrointestinal symptoms and pneumonia severity.

BACKGROUND/AIM: The coronavirus disease 2019 (COVID-19) had become a big threat worldwide. Liver injury is not uncommon in patients with COVID-19, and clarifying its characteristics is needed. This study aimed to identify factors associated with liver injury and to develop a new classification of predictive severity in patients with COVID-19. METHODS: Confirmed patients with COVID-19 (n = 60) were recruited retrospectively from Musashino Red Cross Hospital. The factors of liver injury especially on the elevation of liver enzymes (aspartate aminotransferase [AST] and alanine aminotransferase [ALT]) were analyzed. Grading was assessed according to the Common Terminology Criteria for Adverse Events (CTCAE) version 5.0. RESULTS: During a median hospitalization follow-up of 15 (4-41) days, 51 (85.0%) patients had COVID-19 pneumonia. In clinical courses, oxygenation was needed for 25 (41.6%) patients and intubation was needed for 9 (15.0%) patients. A total of 27 (45.0%) patients had gastrointestinal symptoms (GS), such as appetite loss, diarrhea, and nausea. A logistic regression analysis revealed that C-reactive protein (CRP) at baseline, oxygenation, intubation, and GS were significant factors of liver injury. Based on these results, patients were classified into three groups: group 1, no oxygenation pneumonia; group 2, pneumonia with oxygenation or GS; and group 3, intubation. We classified 25 (41.7%), 26 (43.3%), and 9 (15.0%) patients into mild, moderate, and severe groups, respectively. The peak of AST and ALT levels was significantly stratified with this criteria (mild [median AST, 28 IU/L; median ALT, 33 IU/L], moderate [median AST, 48 IU/L; median ALT, 47.5 IU/L], and severe [median AST, 109 IU/L; median ALT, 106 IU/L]; P<0.001 and P = 0.0114, respectively). CONCLUSION: COVID-19-related liver injury was significantly stratified based on GS and severity of pneumonia.

SARS-CoV-2 Screening Test for Japanese Returnees From Wuhan, China, January 2020.

BACKGROUND: Severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) was found to be the causative microorganism of coronavirus disease 2019 (COVID-19), which started to spread in Wuhan, China. This study was to evaluate the effectiveness of questionnaire, symptoms-based screening, and polymerase chain reaction (PCR) screening of returnees from COVID-19-endemic areas on a chartered flight, to examine the proportion of infected persons and the proportion of asymptomatic persons among infected persons who returned from Wuhan. METHODS: A retrospective cohort study was done in 7 tertiary medical institutions in Japan. A total of 566 Japanese who returned from Wuhan participated in the study. RESULTS: Overall, 11 of the 566 passengers had a positive SARS-CoV-2 PCR result for pharyngeal swabs and 6 were asymptomatic. Only fever differed between SARS-CoV-2-positive and -negative individuals (P < .043). Six of the 11 PCR-positive individuals were asymptomatic; 4 remained positive on day 10, and 1 asymptomatic person tested positive up to day 27. Two of the 11 were negative on the first PCR test and positive on the second. CONCLUSIONS: Our results will be important insights on screening returnees from locked-down cities, as well as providing important data on the proportion of asymptomatic individuals infected with SARS-CoV-2. A 13-day observation period and a second round of PCR may be effective to screen patients, including asymptomatic infections.

Multiple osteolytic bone and lung metastases from prostate cancer including small cell carcinoma with marked increases in CEA and Pro-GRP.

It is known that prostate cancer usually presents as adenocarcinoma, frequently metastasizes to bone, appears osteoblastic on radiographs, and shows elevated PSA. Herein, we describe a case of an 80-year-old man diagnosed with prostate cancer presenting as adenocarcinoma and small cell carcinoma in different areas as well as osteolytic bone metastases in the ilium, right rib, vertebrae, and bilateral femurs with markedly elevated CEA (2391 ng/mL) and Pro-GRP (2610 pg/mL). Occasionally, prostate cancer can appear as osteolytic bone metastases, and in this case, it is possible that the prostate cancer contained small cell carcinoma.

The first case of thrombocytopenia, anasarca, fever, renal impairment or reticulin fibrosis, and organomegaly (TAFRO) syndrome with unilateral adrenal necrosis: a case report.

BACKGROUND: TAFRO syndrome, which was first reported in 2010 in Japan, is a relatively rare disease characterized by thrombocytopenia, anasarca, fever, renal impairment, reticulin fibrosis, and organomegaly. Although this disease is considered similar to multicentric Castleman disease, some of the clinical features, such as thrombocytopenia, are different from typical cases of multicentric Castleman disease. In addition, the etiology of TAFRO syndrome remains unknown and controversial. There have only been a few cases of TAFRO syndrome complicated with adrenal gland lesions, and all of them have had hemorrhagic involvement. CASE PRESENTATION: This report describes the case of a 46-year-old Asian man who presented with fever, epigastric pain, and back pain for 1 month. A computed tomographic scan revealed ascites, mild lymphadenopathy, and left adrenal necrosis without hemorrhage. A blood test showed thrombocytopenia, anemia, and elevated C-reactive protein, alkaline phosphatase, and creatinine levels. Based on the edema, severe thrombocytopenia, fever, reticulin myelofibrosis shown by bone marrow biopsy, mild lymphadenopathy, and progressive renal insufficiency, we diagnosed this patient as having TAFRO syndrome. He was successfully treated by immediate administration of glucocorticoids and tocilizumab. CONCLUSIONS: There have been no previous reports of a case of TAFRO syndrome complicated with adrenal necrosis. Because the biopsy of the left adrenal gland revealed necrosis without any evidence of hemorrhage, we concluded that the unilateral adrenal necrosis in this case was caused by either ischemia from infarction or organomegaly itself under severe hypercytokinemia. This unusual clinical course is useful for further analysis of the etiology of TAFRO syndrome.

Isodicentric Philadelphia chromosome in acute lymphoblastic leukemia with der(7;12)(q10;q10).

We describe here the first case of acute lymphoblastic leukemia (ALL) with an isodicentric Philadelphia [idic(Ph)] chromosome. A 35-year-old man was diagnosed as ALL because of the infiltration of CD10(+)CD19(+)CD33(+)CD34(+) lymphoblasts in the bone marrow and the expression of p190-type BCR/ABL fusion transcript. Chromosome analysis showed 45,XY,der(7;12)(q10;q10),der(9)t(9;22)(q34;q11),idic der(22)t(9;22)(q34;q11). The idic(Ph) chromosome was spindle-shaped and supposed to be formed by two Ph chromosomes joined at their q terminals, whereas idic(Ph) chromosomes in chronic myelogenous leukemia (CML) have been shown to be fused at the satellite regions of p arms. The results indicate that the structure of idic(Ph) chromosomes appears to be different between ALL and CML. The patient did not respond to any chemotherapy and could not achieve remission. This chromosome aberration in ALL may suggest poor prognosis as observed in some cases of CML. Furthermore, considering other three reported cases, der(7;12)(q10;q10) may be one of the recurrent translocations in ALL.

Acquired gain of an X chromosome as the sole abnormality in the blast crisis of chronic neutrophilic leukemia.

Chronic neutrophilic leukemia (CNL) is a rare myeloproliferative disorder characterized by sustained neutrophilic leukocytosis and absence of the Philadelphia chromosome. Most patients with CNL have normal karyotypes, and no specific cytogenetic abnormality has been identified. We report here a patient with CNL that evolved to myeloid blast crisis. A 73-year-old man was admitted to the hospital because of marked leukocytosis (leukocyte count 112.5 x 10(9)/L with 91% segmented neutrophils) and massive hepatosplenomegaly that was diagnosed as CNL with a normal karyotype. After treatment with hydroxyurea for 7 months, the disease progressed to a blast crisis. Bone marrow showed myeloid hyperplasia with 21% myeloblasts, 15% promyelocytes, and marked dysplastic changes of neutrophils. Blastic cells were positive for CD10, CD13, CD14, CD33, CD34, and HLA-DR. Chromosome analysis of the bone marrow cells showed 46,XY,+X in all 20 metaphase spreads. We reviewed 15 cases of CNL terminating in the blast crisis and confirmed that all cases transformed into myeloid crises and had poor prognoses. Furthermore, to our knowledge, this is the first case showing the acquired gain of an extra X chromosome as a sole abnormality in CNL. The gain of an extra X chromosome may play an important role in the progression from chronic phase to the blast crisis of CNL.

New complex t(2;11;17)(p21;q23;q11), a variant form of t(2;11), associated with del(5)(q23q32) in myelodysplastic syndrome-derived acute myeloblastic leukemia.

The t(2;11)(p21;q23) is a rare recurrent aberration observed in myelodysplastic syndrome (MDS) and acute myeloblastic leukemia (AML). It has been suggested that t(2;11) is specifically associated with a deletion of the long arm of chromosome 5 (5q). A 63-year-old man was initially diagnosed as AML with del(5)(q23q32) as a sole abnormality. At relapse, t(2;11;17)(p21;q23;q11) in association with del(5q) appeared in 14 of 20 cells by G-banding. Spectral karyotyping confirmed three derivative chromosomes, der(11)t(2;11), der(17)t(11;17), and der(2)t(2;17). Fluorescence in situ hybridization analysis with a probe for MLL demonstrated that the breakpoint at 11q23 was telomeric to the MLL gene. Nine of 10 reported cases with t(2;11) and del(5q) had MDS including 5q- syndrome and four of them evolved to AML, as observed in the present case. Our results indicated that t(2;11;17) was a secondary genetic change, which appeared during disease progression after del(5q) was observed. Furthermore, considering another reported case, the MLL gene seems to be not involved in the pathogenesis of MDS/AML with t(2;11) and del(5q).

Translocation (8;12)(q13;p13) during disease progression in acute myelomonocytic leukemia with t(11;19)(q23;p13.1).

We report here the first case of acute myelomonocytic leukemia (AMMoL) with both t(8;12)(q13;p13) and t(11;19)(q23;p13.1). A 75-year-old woman was initially diagnosed as having AMMoL with t(11;19) (q23;p13) as a sole abnormality. At the second relapse, G-banding analysis of the bone marrow cells showed 46,XX,t(11;19)(q23;p13)/46,XX,t(8;12)(q13;p13),t(11;19)(q23;p13). Fluorescence in situ hybridization analysis with chromosome-specific painting probes confirmed both the der(8)t(8;12) and the der(12)t(8;12). Reverse transcription-polymerase chain reaction analysis detected the MLL/ELL fusion transcript, indicating that the breakpoint on chromosome 19 was 19p13.1. Leukemic cells at the second relapse were positive for CD2, CD13, CD33, and CD34 but negative for CD14 and HLA-DR. The patient died within 2 months after a subclone with t(8;12)(q13;p13) had appeared. In the literature, t(8;12)(q12;p13) has been observed in two cases of myelodysplastic syndrome and one case of acute myeloblastic leukemia. Our results indicated that t(8;12)(q13;p13) may be one of the recurrent aberrations in myeloid malignancies, although molecular heterogeneity of the breakpoints might exist. Furthermore, it is suggested that t(8;12)(q13;p13) may play an important role in the progression of the disease and lead to the poor prognosis.

[Successful treatment with G-CSF and continuous infusion of low-dose cytarabine and etoposide for therapy-related acute myeloid leukemia developed during chemotherapy for malignant lymphoma].

In March 2000, a 30-year-old Chinese male was initially diagnosed as having non-Hodgkin's lymphoma because of right cervical lymphadenopathy. He had received 8 cycles of chemotherapy including doxorubicin in China. As of February 2001, he was treated in our hospital with the CEPP regimen including etoposide, and was admitted in June 2001 because of leukopenia and thrombocytopenia. Peripheral blood showed hemoglobin 12.7 g/dl, platelets 4.1 x 10(4)/microliter and white blood cells 2300/microliter with 15% blasts. Bone marrow was hypocellular with 48% blasts, which were positive for myeloperoxidase, CD13 and CD33. Chromosome analysis showed 46,XY, t(9;11) (p21;q23) in all 20 metaphase spreads. He was diagnosed as having therapy-related acute myeloblastic leukemia (AML). Because of hypoplastic bone marrow, induction therapy with the CAG regimen including cytarabine, aclarubicin and granulocyte-colony stimulating factor (G-CSF) was started, but no apparent effect was observed. The patient was then treated with the AVG regimen comprising 250 micrograms of G-CSF and continuous infusion with 20 mg of cytarabine and 50 mg of etoposide for 14 days. Complete hematological and cytogenetic remission was achieved after two courses of the AVG regimen. Although it has been shown that the CAG regimen is effective for refractory and/or secondary AML, our results indicate that the AVG regimen should be tried for cases of AML resistant to the CAG regimen.

Infectious mononucleosis accompanied by clonal proliferation of EBV-infected cells and infection of CD8-positive cells.

A 22-year-old male was admitted for a sustained fever of 2 months, lymphadenopathy, and liver dysfunction. Anti-VCA-IgM antibody was positive, with elevated Epstein-Barr virus (EBV)-DNA load in the peripheral blood. Liver biopsy revealed infiltration of CD8-positive and EBV-positive cells. Most peripheral blood mononuclear cells (PBMCs) were also positive for CD8, and showed detectable levels of EBV-DNA. Monoclonal proliferation of EBV-infected cells was detected in the PBMCs by Southern blotting for EBV-terminal repeat (EBV-TR). Although EBV-positive T-cell lymphoproliferative disease (EBV-T-LPD) was suspected, the symptoms spontaneously resolved within 12 months. Anti-VCA-IgM antibody and the clonal band of EBV-TR were negative 1 year after the onset, while anti-EBNA antibody was positive. The final diagnosis was thus confirmed as infectious mononucleosis (IM). Our results indicate that EBV-infected CD8-positive cells and clonal proliferation of EBV-infected cells may be temporally detected in IM. EBV-T-LPDs should be carefully excluded in such cases.

A retrospective cohort study of venous thromboembolism(VTE) in 1035 Japanese myeloma patients treated with thalidomide; lower incidence without statistically significant association between specific risk factors and development of VTE and effects of thromboprophylaxis with aspirin and warfarin.

BACKGROUND: Rate of thalidomide-related VTE and risk factors in Japanese myeloma patients are not clear and effects of thromboprophylaxis remain controvertial. PATIENTS AND METHODS: We retrospectively analyzed a cohort data of registered Japanese myeloma patients treated with thalidomide-based regimens between 2009 and 2010. Primary endpoint was rate of symptomatic VTE. Secondary endpoints were associations between VTE and clinical factors including age, gender, disease characteristics and duration, history of VTE, immobilization, comorbidities, treatment regimens, and laboratory parameters of Hb, leukocyte, platelet and FDP or D-dimer level, and effects of thromboprophylaxis with aspirin or warfarin. Statistical analysis was performed by Fischer's exact test and Cochran-Mantel-Haenszel test to control for confounders, and t test for dichotomous and continuous variables, respectively. RESULTS: 1035 refractory or relapsed myeloma patients were followed up for a median of 112days(range 2-311days), and 14 (1.4%) developed VTE with a median treatment of 31days (range 9-134days) with thalidomide. Treatments with or without other agents lead to similar rates of VTE, 1.7% and 1.1%, respectively (p=0.43) and no specific clinical factors influenced development of VTE. Thromboprophylaxis with aspirin or warfarin did not reduce risk of VTE; VTE with or without aspirin: 1.4% and 1.3% (p=1.00), and warfarin: 2.4% and 1.3% (0.31), respectively. CONCLUSIONS: Rate of VTE is low in Japanese myeloma patients treated with thalidomide. Risk factors and effects of thromboprophylaxis with aspirin or warfarin are not apparent, however, controlled randomized studies of larger scale are needed for statistically valid conclusion.