Pneumocystis jirovecii pneumonia after CD4+ T-cell recovery subsequent to CD19-targeted chimeric antigen receptor T-cell therapy: A case report and brief review of literature.

BACKGROUND: CD19-targeted chimeric antigen receptor (CAR)-T cell therapy involves administration of patient-derived T cells that target B cells, resulting in B-cell depletion and aplasia. In immunity against Pneumocystis jirovecii (Pj), CD4+ T cells and, more recently, B cells, are generally considered important. Antigen presentation by B cells to CD4+ T cells is particularly important. Trimethoprim-sulfamethoxazole (TMP/SMX) for Pj pneumonia (PJP) prophylaxis is generally discontinued when the CD4+ T-cell count is >200/µL. Here we report the first case, to our knowledge, of PJP in a patient with a CD4+ T cell count of >200/µL after CAR-T cell therapy. CASE: A 14-year-old girl developed hemophagocytic lymphohistiocytosis (HLH) after cord blood transplantation (CBT) for relapsed precursor B-cell acute lymphoblastic leukemia (B-ALL). Twenty-one months after CBT, she was diagnosed with combined second relapse in the bone marrow and central nervous system. The patient was treated with CD19-targeted CAR-T cell therapy for the relapse. After CAR-T cell therapy, the patient remained in remission and continued to receive TMP/SMX for PJP prophylaxis. Seven months after CAR-T cell therapy, CD4+ T cells recovered and TMP/SMX was discontinued. The B-cell aplasia persisted. Ten months after CAR-T cell therapy, the patient developed PJP. The patient was also considered to have macrophage hyperactivation at the onset of PJP. Treatment with immunoglobulin, TMP/SMX, and prednisolone was initiated, and the patient's symptoms rapidly ameliorated. CONCLUSION: The patient in the present case developed PJP despite a CD4+ T-cell count of >200/µL after CAR-T cell therapy, probably because of inadequate CD4+ T-cell activation caused by B-cell depletion after CAR-T cell therapy and repeated abnormal macrophage immune responses after CBT. It is important to determine the duration of TMP/SMX for prophylaxis after CAR-T cell therapy according to each case, as well as the CD4+ T-cell count.

Low-cost Methods for Making 3D Fluidic Polymer and Glass Chips Using Metal Templates.

Microfluidics is a rapidly growing field in which small volumes of liquid are moved through channels in a large variety of applications. Fabricating such channels can be expensive. Here, we describe an inexpensive method for making 3D channels in fluidic chips by using a sacrificial template made of coated metal wire or metal tubes. A 3D template is embedded in polymer or glass and then dissolved, leaving channels in the chip, without the need for expensive instruments. By changing the mold, chips of various shapes can be made.

Expression of myostatin in early postnatal mouse masseter and rectus femoris muscles.

AIMS: Myostatin (Mstn) is a member of the transforming growth factor-ß (TGF-ß) family that inhibits muscle differentiation. In this study, we aimed to identify the relationships between Mstn, thyroid hormone receptor alpha (TRα), and myosin heavy chain (MyHC) isoform expression during early postnatal development. METHODS: We investigated the expression of Mstn, TRα, and MyHCs (embryonic, slow, IIa, IIb, and IIx) using quantitative real-time RT-PCR and ELISA (Mstn) in postnatal mouse muscles between day 0 and day 10. We also examined the correlations between Mstn, TR and MyHCs during the early development of mouse masseter muscle (MM) and rectus femoris muscle (RFM). RESULTS: Distinct Mstn mRNA expression patterns were observed in the two muscles despite nearly non-significant changes in the Mstn protein abundance in MM. The expression pattern of the TRα mRNA in the MM differed from that observed in the RFM. The expression of MyHC IIa, IIb and IIx mRNAs increased in the MM and decreased in the RFM from day 0 to day 10, whereas embryonic fiber MyHC mRNA expression was similar in both muscle types. Principal component analysis showed the existence of a correlation between: (1) TRα and MyHC, (2) Mstn and MyHC, and (3) TRα and Mstn in MM. The correlations were different in RFM and MM. Cluster analyses identified the distinct clusters: cluster 1, days 0-4 for the MM and day 0 for the RFM; cluster 2, day 6 for the MM and day 2 for the RFM; and cluster 3, days 8-10 for the MM and days 4-10 for the RFM. CONCLUSIONS: These data suggest that TRα influences MyHC expression in both muscle types. In addition, Mstn has a limited effect in the MM related to the expression of individual MyHCs, as opposed to its role in the RFM, at early postnatal developmental stages. TRα could be involved in regulating both the temporal expression of MyHCs and Mstn at the early postnatal stages in the MM and RFM.

Study of pruritus in chronic hepatitis C patients.

AIM: To investigate the occurrence and severity of pruritus in chronic hepatitis C patients treated with or without interferon (IFN) therapy. METHODS: A total of 89 patients with chronic hepatitis C and 55 control (non-hepatitis) patients were asked to rate their experience of diurnal and nocturnal pruritus in the preceding week using a visual analogue scale (VAS) and a five-point scale, respectively. Blood samples were taken and serum thymus and activation-regulated chemokine (TARC) levels were measured by enzyme-linked immunosorbent assay. RESULTS: A significantly greater proportion of chronic hepatitis C patients experienced nocturnal pruritus compared with control (58.4% vs 5.5%, P < 0.0001). Chronic hepatitis C patients also had more severe pruritus compared with control patients, indicated by the higher mean VAS scores in both the IFN-treated and non-IFN-treated groups. In particular, patients who received combined peginterferon alfa-2b and ribavirin had significantly higher mean VAS scores than those receiving peginterferon alfa-2a or no IFN treatment. Serum TARC levels did not correlate with pruritus scores, and no significant differences in TARC levels were observed between the IFN-treated and non-IFN-treated groups. CONCLUSION: Patients with chronic hepatitis C experience pruritus more than those without. Serum TARC levels do not correlate with pruritus severity in chronic hepatitis C patients.

Measurement of serum branched-chain amino acids to tyrosine ratio level is useful in a prediction of a change of serum albumin level in chronic liver disease.

AIM: In patients with hepatitis C virus (HCV)-associated chronic liver diseases, especially in those with liver cirrhosis, accurate evaluation of their protein nutrition status is very important to improve their quality of life. Whereas the serum albumin level is commonly used to evaluate patients' protein nutrition status, in the present study, the serum amino acid levels were measured, as they also provide valuable information. METHODS: Serum albumin levels and branched-chain amino acids (BCAA) to tyrosine ratio (BTR) were determined in 447 patients with HCV-associated chronic liver diseases (313 with chronic hepatitis and 134 with liver cirrhosis). RESULTS: Chronic hepatitis progressed to liver cirrhosis, serum albumin and serum BTR levels decreased significantlyas chronic hepatitis progressed to liver cirrhosis. Hypoalbuminemia was significantly more common in patients with liver cirrhosis than in those with chronic hepatitis; however, the incidence of an amino acid imbalance was significantly higher than that of hypoalbuminemia in patients with liver cirrhosis. The presence of an amino acid imbalance was associated with a reduction in the serum albumin level 1 year later. CONCLUSIONS: It is important to evaluate serum albumin levels and the BTR in patients with HCV-associated chronic liver diseases.

Effect of symptomatic gastroesophageal reflux disease on quality of life of patients with chronic liver disease.

AIM: Reflux esophagitis is becoming increasingly more prevalent in Japan. It has been noted that symptomatic gastroesophageal reflux disease (GERD) and chronic liver disease may adversely affect patients' quality of life. METHODS: In the present study, 238 chronic liver disease patients (151 patients with chronic hepatitis and 87 patients with liver cirrhosis) were enrolled. The diagnosis of GERD was made based on the Quality-of-Life and Utility Evaluation Survey Technology questionnaire. Health-related quality of life was evaluated using the Short Forum 36 questionnaire. RESULTS: Symptomatic GERD was present in 31.8% (48/151) of patients with chronic hepatitis and 36.8% (32/87) of patients with liver cirrhosis. Among the chronic hepatitis group, compared to the GERD-negative group, the GERD-positive group had significantly lower scores in six domains, including "rolelimitation due to physical problem", "bodily pain", "general health perception", "vitality", "role limitation due to emotional problem", and "mental health". Among the cirrhotic group, compared to the GERD-negative group, the GERD-positive group had significantly lower scores in the "role limitation due to emotional problem" domain. Significant improvement in the "physical functioning", "bodily pain", and "general health perception" domain scores was noted in chronic hepatitis patients treated with rabeprazole. CONCLUSION: The QOL of chronic liver disease patients with symptomatic GERD was impaired.

Protective effect of n-3 polyunsaturated fatty acid on primary culture of rat hepatocytes.

BACKGROUND AND AIM: Recently, we reported on the beneficial clinical effects of eicosapentaenoic acid (EPA) in patients with primary biliary cirrhosis (PBC) who were unresponsive to ursodeoxycholic acid (UDCA). In this study we examined the effect of EPA on rat hepatocytes in primary culture. METHODS: Hepatocytes were isolated from rat liver by perfusion of collagenase and cultured with or without EPA. Cell damage induced by chenodeoxycholic acid (CDCA) was assessed by WST-8 assay and lactate dehydrogenase (LDH) release. PGE(2) and LTB(4) concentrations in the culture medium were measured by enzyme-linked immunosorbent assay (ELISA). cDNA was made from total RNA that was extracted from hepatocytes, and TaqMan polymerase chain reaction (PCR) was performed to assess the expression of CuZn and Mn superoxide dismutase (SOD) mRNA. RESULTS: When rat hepatocytes were cultured in the presence of EPA, the damage caused by CDCA was significantly decreased compared with cells cultured without EPA. Cytotoxicity significantly decreased in the presence of EPA. Furthermore, SOD mRNA expression was increased by adding EPA. These findings indicated that EPA protects cells by scavenging superoxide radicals ((*)O(2-)) mediated by SOD production. CONCLUSION: EPA has a direct protective effect on rat hepatocytes, which is in agreement with the clinical efficacy of EPA in PBC patients.

The impact of visceral fat on multiple risk factors and carotid atherosclerosis in chronic haemodialysis patients.

BACKGROUND: There has been recent interest in the importance of visceral fat (VF) for the development of atherosclerosis. The purpose of this study was to examine associations between VF and multiple risk factors as well as the prevalence of carotid atherosclerosis in chronic haemodialysis patients. METHODS: We classified 77 non-diabetic haemodialysis patients into 'low VF', 'middle VF' and 'high VF' groups after determining VF area using computed tomography. Systemic atherosclerosis was assessed from intima-media thickness (IMT), plaque score (PS) and stiffness parameter beta (stiffness-beta) measured by high-resolution B-mode ultrasonography. RESULTS: Compared with the low VF group, the high VF group exhibited (i) significantly higher fasting plasma insulin (11.0 +/- 6.8 vs 7.1 +/- 2.9 micro U/ml, P = 0.0061); (ii) significantly higher plasma triglycerides (141.8 +/- 94.0 vs 86.5 +/- 32.5 mg/dl, P = 0.0032); and (iii) significantly lower plasma high-density lipoprotein cholesterol (42.1 +/- 14.5 vs 53.0 +/- 15.7mg/dl, P = 0.0134). Moreover, the high VF group had a higher prevalence and extent of carotid atherosclerosis: IMT was 0.69 +/- 0.13 vs 0.61 +/- 0.12 mm (P = 0.0239), PS was 4.8 +/- 3.2 vs 2.4 +/- 3.6 (P = 0.0236) and stiffness-beta was 11.4 +/- 3.1 vs 8.5 +/- 3.0 (P = 0.0082) in the high and low VF groups, respectively. CONCLUSION: We show that VF is associated with the prevalence of carotid atherosclerosis as well as with hyperinsulinaemia and lipid abnormalities in chronic haemodialysis patients.

Clinical significance of KL-6, a marker of interstitial pneumonia, in cases of HCV-associated chronic liver disease.

OBJECTIVE: Various antiviral therapies, including interferon therapy, are being conducted to treat chronic hepatitis C and suppress the onset of hepatocellular carcinoma. However, interstitial pneumonia is beginning to be recognized as one of the adverse reactions of this therapy, and is one of the complications associated with chronic hepatitis. Therefore, we measured the level of KL-6, an interstitial pneumonia marker, in patients with HCV-associated chronic disease, and then determined the possibility of utilizing serum KL-6 as a predictive factor for interstitial pneumonia and the clinical significance of KL-6 in HCV-associated chronic disease. SUBJECTS AND METHODS: The subjects were 308 patients who were diagnosed with chronic liver disease through biochemical blood tests and abdominal diagnostic imaging. All patients tested positive for either the HCV antibody or HCV-RNA, and those who were suspected of having autoimmune hepatitis were excluded. One hundred eighty-five patients had chronic hepatitis (average age: 56 +/- 14 years), while 123 patients had liver cirrhosis (average age: 64 +/- 9 years). The purpose of the present study was explained to every subject, and informed consent was obtained. RESULTS: The mean KL-6 level for chronic hepatitis patients without interstitial pneumonia was 283.5 +/- 131.4 U/ml, while that for cirrhotic patients without interstitial pneumonia was significantly higher, at 377.6 +/- 212.1 U/ml (p<0.0001). In addition, with a cut-off value of 500 U/ml, the ratio of high KL-6 for the chronic hepatitis patients was 5.41% (10/185), while that for the cirrhotic patients was significantly higher, at 20.33% (25/123) (p<0.0001). Furthermore, the mean KL-6 level for patients with a serum hyaluronic acid level of less than 100 ng/ml was 258.4 +/- 124.6 U/ml, while that for patients with a serum hyaluronic acid level of 100 ng/ml or above was significantly higher, at 381.0 +/- 197.3 U/ml (p<0.0001). CONCLUSION: Although KL-6 is a marker of interstitial pneumonia, the results of the present study suggest that, in HCV-associated chronic disease, this marker reflects hepatic fibrosis better than pulmonary fibrosis.