Development of a rapid and novel diagnostic technique for cardiac amyloidosis using Raman spectroscopy.

Although the prognosis of cardiac amyloidosis has improved with the development of therapies, the time required for disease typing remains a critical issue. We investigated the potential of Raman spectroscopy for the more rapid diagnosis and typing of cardiac amyloidosis. Heart biopsies were collected from patients with the AL (4) and ATTR (4) types of cardiac amyloidosis, and tissue sections were subjected to Raman microscopy. A principal component analysis (PCA) of spectral data was performed and receiver operating characteristic (ROC) curves were created to confirm the accuracy of discriminating between amyloid-deposition and non-deposition sites, and between AL and ATTR. The steep peak at 1680 cm-1, reflecting the β-sheet structure, was useful for detecting the amyloid-deposition region. By restricting the spectral analysis to amyloid-deposition sites, AL and ATTR were discriminated by principal components with a characteristic broad peak at 1520-1540 cm-1, which was also observed in the Raman spectrum of AL, but not ATTR. The area under ROC curve discriminating AL and ATTR was 0.78. PCA of the Raman spectra of cardiac biopsies has the potential not only to detect amyloid-deposition sites in tissue but also to rapidly discriminate between the AL and ATTR types of cardiac amyloidosis.


Detection of Hematological Malignancies Using N-NOSE (Nematode-NOSE).

Hematological malignancies often lack defined risk factors and present with non-specific symptoms, underscoring the urgent need for simple and reliable detection methods. To address this challenge, Hirotsu et al. innovated N-NOSE, a novel, non-invasive cancer screening test that utilizes the chemotaxis response of the nematode Caenorhabditis elegans to detect tumor-related odors in urine. In this clinical study, we assessed the performance of N-NOSE in patients with various hematological malignancies at diagnosis and during treatment. Urine samples were collected from 30 healthy individuals and 89 patients, including those with leukemia (n = 13), malignant lymphoma (n = 53), multiple myeloma (n = 15), primary AL amyloidosis (n = 3), Waldenström's macroglobulinemia (n = 2), myelodysplastic syndrome (n = 2), and blastic plasmacytoid dendritic cell neoplasm (n = 1). Based on the optimal cut-off values in detecting hematological malignancies, N-NOSE demonstrated high positivity rates in treatment-naïve patients: leukemia and multiple myeloma were very high (over 90%), whereas malignant lymphoma was slightly lower than 80%. In the small subset of malignant lymphoma patients who tested N-NOSE-negative, confounding factors included steroid administration and hemodialysis. Importantly, no significant correlation emerged between N-NOSE index values and baseline characteristics or comorbidities other than the presence of cancer. Moreover, in all 32 patients who achieved clinical response following chemotherapy, the N-NOSE index declined, reflecting disease status. These findings highlight N-NOSE's strong potential as a sensitive, non-invasive screening tool for hematological malignancies-particularly multiple myeloma-and support its use in initial detection and monitoring of therapeutic response.


False-Positive Galactomannan Test Results in Multiple Myeloma.

Invasive pulmonary aspergillosis (IA) is a common infectious disease in patients with hematological diseases. The prevention, early detection, and establishment of treatment strategies for IA are important. The serum galactomannan antigen (GM) mycological test for IA diagnosis, included in the mycology criteria of the European Organization for Research and Treatment of Cancer-Invasive Fungal Infections Cooperative Group/National Institute of Allergy and Infectious Diseases Mycosis Study Group (EORTC/MSG), is widely used because of its high sensitivity and specificity. However, false-positive results are a concern.We retrospectively analyzed all GM tests performed at our department in the clinical practice setting between April 2003 and January 2012.Of the 330 cases and 2155 samples analyzed, 540 (25%) were positive (0.5). Among the underlying diseases, positivity rates were the highest for multiple myeloma (MM), with 61.3%. By type, positivity rates for IgG, IgA, Bence-Jones protein, and IgD were 71.7%, 33.3%, 57.1%, and 34.6%, respectively. Seventeen out of eighteen cases that were GM-positive at MM diagnosis were false positives, according to the 2008 EORTC/MSG criteria. The IgG and GM values were not directly correlated. Of the seventeen false-positive cases identified, two developed IA during anti-myeloma treatments, and GM values did not become negative during the treatment in most cases.Although subclinical IA may be included in a higher GM index, the results may be prone to false positives; particularly in IgG-type MM, the results should thus be interpreted cautiously.


Community medical care faces challenges such as the uneven distribution of physicians, fluctuating medical demand, and the increased relative demand for physicians due to the increased specialization of medical fields. Hematologists primarily address issues, such as abnormal blood cell counts, lymphadenopathy, persistent fever, and coagulation abnormalities, which indicate hematological diseases. However, the emphasis on treating hematologic diseases within community medicine remains relatively low. Accurate and reliable differentiation and identification of hematological diseases with the cooperation of laboratory technicians and support of artificial intelligence is necessary. Significant advances have been made in the treatment of hematological diseases; however, small community hospitals often lack access to these treatments and are unable to conduct clinical trials that require specialized equipment such as for chimeric antigen receptor T-cell therapy. In the future, hematologists will need to focus on developing their careers within the community and further optimizing their practice to enhance patient care. J. Med. Invest. 72 : 21-25, February, 2025.

The Journal of Medical Investigation : JMI


Elotuzumab-mediated ADCC with Th1-like Vγ9Vδ2 T cells to disrupt myeloma-osteoclast interaction.

Multiple myeloma (MM) cells and osteoclasts (OCs) activate with each other to cause drug resistance. Human Th1-like Vγ9Vδ2 (γδ) T cells, important effectors against tumors, can be expanded and activated ex vivo by the aminobisphosphonate zoledronic acid in combination with IL-2. We previously reported that the expanded γδ T cells effectively targeted and killed OCs as well as MM cells. Because the expanded γδ T cells expressed CD16 on their surface, we investigated the utilization of the expanded γδ T cells for antibody-dependent cellular cytotoxicity (ADCC). Although the expanded γδ T cells alone induced cell death in MM cell lines, the addition of the anti-SLAMF7 monoclonal antibody elotuzumab (ELO) further enhanced their cytotoxic activity only against SLAMF7-expressing MM cell lines and primary MM cells. Intriguingly, ELO was also able to enhance γδ T cell-induced cell death against OCs cultured alone, and against both MM cells and OCs in their coculture settings. SLAMF7 was found to be highly expressed in OCs differentiated in vitro from monocytes by receptor activator of nuclear factor-κ B ligand and M-CSF, although monocytes only marginally expressed SLAMF7. These results demonstrate that SLAMF7 is highly expressed in both MM cells and OCs, and that the ex vivo-expanded γδ T cells can exert ELO-mediated ADCC against SLAMF7-expressing MM cells and OCs besides their direct cytotoxic activity. Further study is warranted for the innovative utilization of γδ T cells.


Raman Microspectroscopy for Label-Free Diagnosis of Amyloid Light-chain Amyloidosis in Various Organs


Phase Angle and Extracellular Water-to-Total Body Water Ratio Measured by Bioelectrical Impedance Analysis


Acute myeloid leukemia developed through myeloproliferative features during immunosuppressive therapy for juvenile idiopathic arthritis.

A 17-year-old male with thrombocytosis and exacerbation of arthralgia during intensified immunosuppressive therapy with tocilizumab, prednisolone, and methotrexate for juvenile idiopathic arthritis (JIA) was referred to our department. Bone marrow examination revealed myelodysplastic syndrome/myeloproliferative neoplasm, unclassifiable (MDS/MPN-U). Peripheral myeloblasts disappeared temporarily after discontinuation of tocilizumab but progressed to acute myeloid leukemia six months after the development of MDS/MPN-U. The patient sustained complete remission after unrelated bone marrow stem cell transplantation, followed by chemotherapy. The arthralgia also improved after chemotherapy. The possibility of developing malignancies during immunosuppressive therapy in patients with JIA should be considered. J. Med. Invest. 71 : 335-339, August, 2024.


Therapeutic efficacy of the resorcylic acid lactone LL-Z1640-2 for adult T-cell leukaemia/lymphoma

Adult T-cell leukaemia/lymphoma (ATL) remains incurable. The NF-κB and interferon regulatory factor 4 (IRF4) signalling pathways are among the critical survival pathways for the progression of ATL. TGF-β-activated kinase 1 (TAK1), an IκB kinase-activating kinase, triggers the activation of NF-κB. The resorcylic acid lactone LL-Z1640-2 is a potent irreversible inhibitor of TAK1/extracellular signal-regulated kinase 2 (ERK2). We herein examined the therapeutic efficacy of LL-Z1640-2 against ATL. LL-Z1640-2 effectively suppressed the in vivo growth of ATL cells. It induced in vitro apoptosis and inhibited the nuclear translocation of p65/RelA in ATL cells. The knockdown of strongly induced ATL cell death while downregulating MYC. LL-Z1640-2 as well as the NF-κB inhibitor BAY11-7082 decreased the expression of IRF4 and MYC at the protein and mRNA levels, indicating the suppression of the NF-κB-IRF4-MYC axis. The treatment with LL-Z1640-2 also mitigated the phosphorylation of p38 MAPK along with the expression of CC chemokine receptor 4. Furthermore, the inhibition of STAT3/5 potentiated the cytotoxic activity of LL-Z1640-2 against IL-2-responsive ATL cells in the presence of IL-2. Therefore, LL-Z1640-2 appears to be an effective treatment for ATL. Further studies are needed to develop more potent compounds that retain the active motifs of LL-Z1640-2.


Novel antimyeloma therapeutic option with inhibition of the HDAC1-IRF4 axis and PIM kinase

Multiple myeloma (MM) preferentially expands and acquires drug resistance in the bone marrow (BM). We herein examined the role of histone deacetylase 1 (HDAC1) in the constitutive activation of the master transcription factor IRF4 and the prosurvival mediator PIM2 kinase in MM cells. The knockdown or inhibition of HDAC1 by the class I HDAC inhibitor MS-275 reduced the basal expression of IRF4 and PIM2 in MM cells. Mechanistically, the inhibition of HDAC1 decreased IRF4 transcription through histone hyperacetylation and inhibiting the recruitment of RNA polymerase II at the IRF4 locus, thereby reducing IRF4-targeting genes, including PIM2. In addition to the transcriptional regulation of PIM2 by the HDAC1-IRF4 axis, PIM2 was markedly upregulated by external stimuli from BM stromal cells and interleukin-6 (IL-6). Upregulated PIM2 contributed to the attenuation of the cytotoxic effects of MS-275. Class I HDAC and PIM kinase inhibitors cooperatively suppressed MM cell growth in the presence of IL-6 and in vivo. Therefore, the present results demonstrate the potential of the simultaneous targeting of the intrinsic HDAC1-IRF4 axis plus externally activated PIM2 as an efficient therapeutic option for MM fostered in the BM.


Systemic amyloidosis associated with non-IgM type paraprotein with lymphoplasmacytic lymphoma


[Adult T-cell leukemia/lymphoma with multiple intracranial masses and CMV and HHV-6 reactivation at initial presentation].

初発時にCMVとHHV-6の再活性化と頭蓋内多発腫瘤を認めた成人T細胞白血病リンパ腫

(ATLL cells were 79% in flow cytometry), and the protein level was 244 mg/dl; moreover, the examination revealed a positive result for human herpesvirus 6 DNA. Despite herpesvirus genus treatment and modified LSG15 therapy combined with intrathecal chemotherapy, the patient became comatose and died on day 21 of hospitalization. A better understanding of the pathogenesis of ATLL, and the involvement with the central nervous system is needed along with the development of standard treatment.

The Japanese Journal of Clinical Hematology


First reported case of Lachnoanaerobaculum gingivalis bacteremia in an acute myeloid leukemia patient with oral mucositis during high dose chemotherapy.

During chemotherapy in patients with oral mucositis, we should consider the possibility of L. gingivalis bacteremia.


Allogeneic haematopoietic stem cell transplantation and patient falls: impact of lower extremity muscle strength.

Pretransplant LEMS was a significant predictor of post-transplant falls. The results of this study may help to prevent falls in patients undergoing allo-HSCT.


Allogeneic haematopoietic stem cell transplantation-clinical outcomes: impact of leg muscle strength.

Pre-transplant LEMS was a significant factor in predicting OS and NRM.


Artifactual prolongation of the activated partial thromboplastin time by amikacin or gentamicin with ellagic acid, but not silica activated reagent.

International Journal of Laboratory Hematology


TAK1 is a pivotal therapeutic target for tumor progression and bone destruction in myeloma

Along with the tumor progression, the bone marrow microenvironment is skewed in multiple myeloma (MM), which underlies the unique pathophysiology of MM and confers aggressiveness and drug resistance in MM cells. TGF-β-activated kinase-1 (TAK1) mediates a wide range of intracellular signaling pathways. We demonstrate here that TAK1 is constitutively overexpressed and phosphorylated in MM cells, and that TAK1 inhibition suppresses the activation of NF-κB, p38MAPK, ERK and STAT3 to decrease the expression of critical mediators for MM growth and survival, including PIM2, MYC, Mcl-1, IRF4, and Sp1, along with a substantial reduction in the angiogenic factor VEGF in MM cells. Intriguingly, TAK1 phosphorylation was also induced along with upregulation of vascular cell adhesion molecule-1 (VCAM-1) in bone marrow stromal cells (BMSCs) in cocultures with MM cells, which facilitated MM cell-BMSC adhesion while inducing IL-6 production and receptor activator of nuclear factor κ-Β ligand (RANKL) expression by BMSCs. TAK1 inhibition effectively impaired MM cell adhesion to BMSCs to disrupt the support of MM cell growth and survival by BMSCs. Furthermore, TAK1 inhibition suppressed osteoclastogenesis enhanced by RANKL in cocultures of bone marrow cells with MM cells, and restored osteoblastic differentiation suppressed by MM cells or inhibitory factors for osteoblastogenesis overproduced in MM. Finally, treatment with the TAK1 inhibitor LLZ1640-2 markedly suppressed MM tumor growth and prevented bone destruction and loss in mouse MM models. Therefore, TAK1 inhibition may be a promising therapeutic option targeting not only MM cells but also the skewed bone marrow microenvironment in MM.


Synergistic targeting of Sp1, a critical transcription factor for myeloma cell growth and survival, by panobinostat and proteasome inhibitors.

Panobinostat, a pan-deacetylase inhibitor, synergistically elicits cytotoxic activity against myeloma (MM) cells in combination with the proteasome inhibitor bortezomib. Because precise mechanisms for panobinostat's anti-MM action still remain elusive, we aimed to clarify the mechanisms of anti-MM effects of panobinostat and its synergism with proteasome inhibitors. Although the transcription factor Sp1 was overexpressed in MM cells, the Sp1 inhibitor terameprocol induced MM cell death in parallel with reduction of IRF4 and cMyc. Panobinostat induced activation of caspase-8, which was inversely correlated with reduction of Sp1 protein levels in MM cells. The panobinostat-mediated effects were further potentiated to effectively induce MM cell death in combination with bortezomib or carfilzomib even at suboptimal concentrations as a single agent. Addition of the caspase-8 inhibitor z-IETD-FMK abolished the Sp1 reduction not only by panobinostat alone but also by its combination with bortezomib, suggesting caspase-8-mediated Sp1 degradation. The synergistic Sp1 reduction markedly suppressed Sp1-driven prosurvival factors, IRF4 and cMyc. Besides, the combinatory treatment reduced HDAC1, another Sp1 target, in MM cells, which may potentiate HDAC inhibition. Collectively, caspase-8-mediated post-translational Sp1 degradation appears to be among major mechanisms for synergistic anti-MM effects of panobinostat and proteasome inhibitors in combination.


Expansion of Th1-like V9V2T cells by new-generation IMiDs, lenalidomide and pomalidomide, in combination with zoledronic acid.


[A case of peritonitis carcinomatosa from goblet cell carcinoid of the appendix treated by intraperitoneal paclitaxel and systemic S-1 chemotherapy].

Goblet cell carcinoid of the appendix is a rare neoplasm and clinically tends to take a malignant course. Most cases are young and early stage, and the surgical strategy is available. But appropriate chemotherapy for inoperable cases with peritoneal dissemination is not established. A 77-year-old woman with a past history of appendectomy was admitted to our hospital complaining of abdominal fullness. Abdominal computed tomography showed massive ascites and slight contrast enhancement of appendix. A tumor was found by colonoscopic examination at the orifice of vermiform and was diagnosed pathologically as goblet cell carcinoid of the appendix. Laparoscopy showed multiple peritoneal dissemination. We performed intraperitoneal paclitaxel(PTX)administration at 70 mg/m(2) week without any resection of the tumor. Ascites were reduced immediately, but drug-induced interstitial pneumonia occurred due to PTX. After steroid therapy, we switched to systemic S-1 therapy. For about one year, her tumor was controlled but became worse thirteen months after diagnosis and died. It is thought that intraabdominal paclitaxel administration and systemic S-1 therapy can be one of appropriate forms of chemotherapy for inoperable peritoneal carcinomatosis from goblet cell carcinoid of appendix.

Japanese Journal of Cancer and Chemotherapy


[A long-surviving patient with lung pleomorphic carcinoma treated with postoperative carboplatin and paclitaxel combination chemotherapy].

We presented the case of a 46-year-old man with no medical or family history but with a history of smoking 3 packs of cigarettes per day for the past 25 years. He was admitted to our hospital due to hemoptysis. Chest computed tomography revealed a tumor of right upper lung and interstitial pneumonia in the surrounding lung parenchyma. He was operated upon and diagnosed with stage IIB pleomorphic carcinoma of the lung with invasion of the chest wall. He underwent three courses of postoperative carboplatin (CBDCA) (area under the curve 5 on day 1, every 3 weeks and paclitaxel(PTX) (200 mg/m(2); day 1, every 3 weeks) combination chemotherapy. No recurrence was observed for a period of 760 days after the operation. According to previous reports, lung pleomorphic carcinoma is aggressive and has a poor prognosis. Further, the significance of chemotherapy in the management of this disease has not been established. Postoperative combination chemotherapy of CBDCA and PTX may result in a good prognosis for this disease.