Pathogen Profiles in Outpatients with Non-COVID-19 during the 7th Prevalent Period of COVID-19 in Gunma, Japan.

The identification of pathogens associated with respiratory symptoms other than the novel coronavirus disease 2019 (COVID-19) can be challenging. However, the diagnosis of pathogens is crucial for assessing the clinical outcome of patients. We comprehensively profiled pathogens causing non-COVID-19 respiratory symptoms during the 7th prevalent period in Gunma, Japan, using deep sequencing combined with a next-generation sequencer (NGS) and advanced bioinformatics technologies. The study included nasopharyngeal swabs from 40 patients who tested negative for severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) using immuno-chromatography and/or quantitative reverse transcription polymerase chain reaction (qRT-PCR) methods. Comprehensive pathogen sequencing was conducted through deep sequencing using NGS. Additionally, short reads obtained from NGS were analyzed for comprehensive pathogen estimation using MePIC (Metagenomic Pathogen Identification Pipeline for Clinical Specimens) and/or VirusTap. The results revealed the presence of various pathogens, including respiratory viruses and bacteria, in the present subjects. Notably, human adenovirus (HAdV) was the most frequently detected virus in 16 of the 40 cases (40.0%), followed by coryneforms, which were the most frequently detected bacteria in 21 of the 40 cases (52.5%). Seasonal human coronaviruses (NL63 type, 229E type, HKU1 type, and OC43 type), human bocaviruses, and human herpesviruses (human herpesvirus types 1-7) were not detected. Moreover, multiple pathogens were detected in 50% of the subjects. These results suggest that various respiratory pathogens may be associated with non-COVID-19 patients during the 7th prevalent period in Gunma Prefecture, Japan. Consequently, for an accurate diagnosis of pathogens causing respiratory infections, detailed pathogen analyses may be necessary. Furthermore, it is possible that various pathogens, excluding SARS-CoV-2, may be linked to fever and/or respiratory infections even during the COVID-19 pandemic.

Influence of CYP3A5 and ABCB1 polymorphisms on the pharmacokinetics of vincristine in adult patients receiving CHOP therapy.

PURPOSE: This study aims to clarify the impact of CYP3A5 and ABCB1 polymorphisms on the pharmacokinetics of vincristine (VCR) in adult patients receiving CHOP therapy. METHODS: Plasma samples were collected immediately after the end of VCR administration and at 1.5, 2.5, 3.5, 5.5, 9.5, 13.5, and 25.5 h after the start of administration. Areas under the plasma concentration-time curves of VCR in the elimination phase (AUC1.5-25.5) were calculated using the linear trapezoidal rule. Half-lives of VCR during the early phase (1.5-5.5 h) and terminal phase (5.5-25.5 h; t1/2γ) were determined according to the log-linear regression of the concentration-time data for at least 3 sampling points. RESULTS: A total of 41 adult patients were enrolled in this study. The median t1/2γ and AUC1.5-25.5 were significantly longer and higher in CYP3A5 non-expressers (CYP3A5*3/*3) than in CYP3A5 expressers (CYP3A5*1/*1 or *1/*3) (21.3 vs 13.8 h, P = 0.005 and 35.5 vs 30.0 ngï½¥h/mL, P = 0.006, respectively). Conversely, there were no significant differences in pharmacokinetic parameters among the ABCB1 c.1236C>T, c.2677G>A/T, c.3435C>T genotype groups. A stepwise selection multiple linear regression analysis showed that the dose of VCR administered and CYP3A5 non-expresser status were independent factors influencing the AUC1.5-25.5 (partial R2 = 0.212, P = 0.002 and partial R2 = 0.143, P = 0.010, respectively). CONCLUSION: The CYP3A5*3 polymorphism was found to be an indicator for predicting exposure to VCR in adult patients receiving CHOP therapy. This information may be useful for the individualization of VCR dosages.

[Four Cases Who Experienced Extravasation of Anthracyclines and Had Dexrazoxane Therapy].

We analyzed 4 cases who experienced extravasation of anthracyclines and had dexrazoxane therapy in our hospital. Concerned drugs were 2 adriamycin and 2 amrubicin cases and all cases received steroid ointment therapy, and no cases showed severe condition such as skin ulcer. As dexrazoxane is known to enhance bone marrow suppression of anti-cancer drugs, the nadir of neutropenia and thrombocytopenia was observed from day 10 to 17 in our cases. We made a domestic manual and have used in various professionals. Dexrazoxane would contribute to the reduction of skin damage due to extravasation if we could manage bone marrow suppression successfully.

Enteral lorlatinib after alectinib as a treatment option in anaplastic lymphoma kinase-positive non-small cell lung cancer with triple problems: carcinomatous meningitis, poor performance status, and dysphagia-a case report.

Alectinib treatment is effective in patients with anaplastic lymphoma kinase (ALK) gene rearrangement-positive non-small cell lung cancer (NSCLC; hereafter ALK-positive NSCLC) who exhibit central nervous system (CNS) relapse and poor performance status (PS). Lorlatinib treatment is effective upon failure of other ALK inhibitor-based treatments. However, much remains unknown about the efficacy of lorlatinib in patients with ALK-positive NSCLC, who have triple problems, carcinomatous meningitis, poor PS, and dysphagia, after alectinib treatment. Here, we report the remarkable response of a 73-year-old patient with ALK-positive NSCLC showing carcinomatous meningitis due to CNS metastases, poor PS, and dysphagia to lorlatinib. Lorlatinib administration through a nasogastric tube alleviated complications related to consciousness within three days, and the patient survived for 16 months after CNS relapse. Lorlatinib could be a treatment option for patients with ALK-positive NSCLC showing carcinomatous meningitis, poor PS, and dysphagia upon failure of other ALK inhibitor-based treatments.

Evaluation for pharmacokinetic exposure of cytotoxic anticancer drugs in elderly patients receiving (R-)CHOP therapy.

(R-)miniCHOP therapy, which delivers approximately half-doses of the (R-)CHOP regimen, has shown efficacy and safety in patients who are more than 80 years old. This study aimed to compare the area under the plasma concentration-time curves (AUCs) of vincristine (VCR), doxorubicin (DXR), and cyclophosphamide (CPA) between (R-)CHOP and (R-)miniCHOP regimens. The AUCs were compared between patients aged 65-79 years receiving (R-)CHOP therapy and those aged 80 years and older receiving (R-)miniCHOP therapy. Age was not an independent variable for predicting the dose-adjusted AUCs (AUC/Ds) of cytotoxic anticancer drugs. The median AUCs of DXR and CPA were significantly smaller in the (R-)miniCHOP group than in the (R-)CHOP group (168.7 vs. 257.9 ng h/mL, P = 0.003, and 219.9 vs. 301.7 µg h/mL, P = 0.020, respectively). The median AUCs of VCR showed the same trend but the difference was not significant (24.83 vs. 34.85 ng h/mL, P = 0.135). It is possible that the AUCs of VCR, DXR, and CPA in patients aged 80 years and older receiving (R-)miniCHOP therapy may be lower than those in patients 65-79 years old receiving (R-)CHOP therapy.

[A Case of Non-Resectable Pancreatic Cancer with Multiple Hepatic Metastases Treated with FOLFIRINOX Therapy and Conversion Surgery].

A 50-year-old woman was diagnosed as having pancreatic head cancer with multiple hepatic metastases. FOLFIRINOX therapy was initiated. After completing 18 courses of therapy, partial remission(PR)was achieved based on images, and surgery was then planed. The subtotal stomach-preserving pancreaticoduodenectomy and hepatic S7 partial resection were performed. Macroscopically, complete resection was achieved. Regarding pathological findings of the primary lesion and hepatic metastatic lesions, fibrous formation and hyalinizing condition induced by chemotherapy were noted; moreover, complete disappearance of cancer cells was detected. However, metastasis of poorly differentiated adenocarcinoma was detected in 12b lymph node tissue. One month after the surgery, postoperative adjunctive chemotherapy with S-1 was initiated. However, new hepatic metastasis was detected 3 months after the surgery. Although recurrence treatment was initiated, the disease progressed, and the patient died 11 months after the surgery.

[Analysis of the Patient Factors in the Difference between Adverse Effect Evaluation and Distress Evaluation in Cancer Patients Receiving Chemotherapy].

A safety evaluation of chemotherapy is performed by CTCAE. It is the evaluation by health care workers, and distress evaluation by patient himself is not included in it. Therefore, a health care worker underestimates patients' distress. This study was carried out to identify the patients' characteristics underlying differences between safety evaluation and distress evaluation. The patients who met the criteria were 72 in number. They were divided into 2 groups. Group A(17 patients)included patients who demonstrated difference between safety evaluation and distress evaluation. Group B(55 patients)included patient who did not show difference between safety evaluation and distress evaluation. The patients who visited a hospital were evaluated for QOL, depression screening, CTCAE(safety evaluation), and PRO-CTCAE(distress evaluation). A meaningful difference was observed between depression screening, QOL-ACD(physical status, psychological status, face scale, and total), and the number of items of side effect by PRO-CTCAE through a univariate analysis. A meaningful difference was observed for QOL-ACD(physical)in logistic regression analysis(odds ratio=1.47, p=0.013). It is suggested that having physical distress reflected by the QOL evaluation before chemotherapy results in the difference between safety evaluation and distress evaluation.

[Retrospective Analysis of Weekly Paclitaxel Chemotherapy for Gemcitabine- and S-1-Resistant Pancreatic Cancer].

Chemotherapy for unresectable pancreatic cancer has moved from using gemcitabine (GEM) and/or S-1 to using 5- fluorouracil plus Leucovorin plus oxaliplatin plus irinotecan (FOLFIRINOX) or GEM and nano albumin-bound paclitaxel (nab- PTA). We administered weekly PTA 80mg/m2 (days 1, 8, 15 every 4 weeks) in 22 patients with both GEM and S-1 resistance before nab-PTX became available through medical insurance in Japan. This regimen was used as a second-line in 3 cases, as the third-line in 14 cases and as a later line in 5 cases. The mean number of chemotherapy courses was 2.7, and the mean dose intensity was 86.1%. Postponement and dose reduction was made in 15 and 5 cases, respectively. The best overall response was 1 PR, 5 SD, 15 PD and 1 NE. The response rate was 4.5%, and disease control rate was 27.3%. The median progression-free survival was 1.7 months, and the median overall survival was 4.6 months. The main adverse events included anorexia, general malaise, and peripheral neurotoxicity and they were tolerable. This study wherein nab-PTX plus GEM was one of the standard therapies, indicated that the PTX alone was effective in pancreatic cancer patients who were resistant to GEM and S-1.

Expression of cancer-associated fibroblast markers in advanced colorectal cancer.

Colorectal cancer is one of the most common causes of mortality from cancer worldwide. Previous studies have demonstrated that cancer-associated fibroblasts (CAFs) promote neoangiogenesis and tumor growth for various tumors. The present study analyzed CAF markers, including α-smooth muscle actin (α-SMA), collagen I, platelet-derived growth factor receptor-ß (PDGFR-ß), and D2-40 (antibody recognizing podoplanin), and vessel markers, including cluster of differentiation (CD)31 and CD34, for 121 advanced colorectal cancer cases using a digital image analyzing technique. The association between CAF markers and vessel markers with clinicopathological factors was investigated. Furthermore, the association between CAF markers with each other, and their association with vessel markers was analyzed. Mean/median expression area of stromal and vessel markers in tumors were collagen I, 26.787%; D2-40, 1.372%; PDGFR-ß, 11.646%; α-SMA-positive and desmin-negative myofibroblasts (α-SMA subtraction), 15.372%; CD31, 3.635%; and CD34, 2.226%. The expression area of α-SMA subtraction was significantly correlated with collagen I (P<0.001, correlation rho=0.509). High levels of α-SMA subtraction (P=0.002), collagen I (P=0.040), and PDGFR-ß (P=0.040) expressions tended to be associated with high venous invasion. D2-40 did not correlate with other CAF and vessel markers. These results indicated that individual CAFs may have different expression patterns, and different strength effects for venous invasion in advanced colorectal cancer stroma.

Prognostic Impact of Extracapsular Lymph Node Invasion and Myofibroblastic Activity in Extrahepatic Bile Duct Cancer.

Extrahepatic bile duct carcinoma is a potentially malignant gastrointestinal lesion. Cancer cells spread via the lymphatic system to regional lymph nodes and help in tumor progression. However, there are no reports on the prognostic impact of extracapsular lymph node invasion and myofibroblastic activity in this cancer. Hence, we classified the histopathologic patterns of lymph nodes into 2 patterns: extracapsular lymph node invasion or not. Based on this, we investigated 32 cases of extrahepatic bile duct cancer with lymph node metastasis and classified 21 cases as positive and 11 cases as negative. The extracapsular lymph node invasion cases were associated with poor disease-free survival and overall survival. The myofibroblast density of the metastatic foci was significantly higher in the extracapsular lymph node invasion cases. This is the first study to demonstrate that extracapsular lymph node invasion cases were associated with poor prognosis and that the myofibroblast distribution contributed to malignancy.

Myofibroblast distribution is associated with invasive growth types of colorectal cancer.

Both the invasive growth types of colorectal cancer (CRC) and the number of myofibroblasts have been associated with histopathological factors such as lymph node and liver metastasis, and local recurrence. However, there are few studies, that have assessed the association between invasive growth type and myofibroblast distribution in CRC. We aimed to evaluate the relationship between the clinicopathological factors of CRC and two invasive growth types, the expanding and infiltrating types. We categorized 150 cases of pT3 CRC into the expanding and infiltrating types and measured the myofibroblast density of three histological layers: the submucosa (SM), the muscularis propria (MP) and the subserosa (SS). We compared these two invasive growth types and analyzed the relationship between clinicopathological factors and myofibroblast density. Myofibroblast density was significantly higher in the infiltrating type than that in the expanding type (P<0.05). In the lymph node metastasis-positive group of the infiltrating type, myofibroblast density in MP was significantly higher than that in the lymph node metastasis-negative group (P<0.001). In the infiltrating type, the group with the higher level of lymphatic invasion had a significantly higher density of myofibroblasts in the MP than the group with the lower level of lymphatic invasion (P<0.01). These results suggest that myofibroblasts participate more in the infiltrating type compared with the expanding type of CRC. It would appear that myofibroblasts present in the MP play an important role in the malignant potential of the infiltrating type compared to the expanding type.

[Withdrawal of Anticancer Therapy in Advanced Disease Because of Side Effects].

Withdrawal of chemotherapy because of side effects may be due to"problems with toxicity"or"patient refusal"."Problems with toxicity"is intended to secure patient safety and decisions about it are primarily made by the oncologist. On the other hand,"patient refusal"is influenced by the degree of distress the patient is experiencing and the patient's preference. Providing sufficient medical information is essential in decision making, because it is a decision made by the patient. In addition, the perception of side effects has changed owing to progress in supportive therapy in recent years. In particular, cancer patients are concerned about changes in appearance. In our study(4/2015 to 3/2016), chemotherapy was withdrawn because of side effects during treatment for solid carcinomas in 8%(4/51)of the patients. Two of these patients experienced liver function disorders, and 2 developed interstitial pneumonitis. Patient refusal was observed in 2%(1/51)of the patients.

[Second-line chemotherapy for advanced pancreatic cancer].

Many investigators have attempted to reach a consensus, though with limited evidence available, on second-line chemotherapy for advanced pancreatic cancer. Specifically, treatment consists of gemcitabine-based regimens and 5-FU-based regimens. The regimen that is not used for first-line chemotherapy will be used as second-line chemotherapy. In Japan, S-1 is frequently used in 5-FU-based regimens. When the FOLFIRINOX regimen is used as first-line chemotherapy, gemcitabine is recommended as second-line chemotherapy. On the other hand, when nab-PTX/gemcitabine is used as first-line chemotherapy, S-1 is recommended as second-line chemotherapy.

Effect of diode laser on proliferation and differentiation of PC12 cells.

This study investigated the effects of diode (GaAlAs) laser irradiation at an effective energy density of 5 or 20 J/cm(2) on cell growth factor-induced differentiation and proliferation in pheochromocytoma cells (PC12 cells), and whether those effects were related to activation of the p38 pathway. Laser irradiation at 20 J/cm(2) significantly decreased the number of PC12 cells, while no difference was seen between the 5 J/cm(2) group and the control group (p<0.05). Western blotting revealed marked expression of neurofilament and ß-tubulin, indicating greater neurite differentiation in the irradiation groups than in the control group at 48 hr. Irradiation also enhanced expression of phospho-p38. The decrease in number of cells after laser irradiation was accelerated by p38 inhibitor, while neurite differentiation was up-regulated by laser irradiation, even when the p38 pathway was blocked. This suggests that laser irradiation up-regulated neurite differentiation in PC12 cells involving p38 and another pathway.

[Evaluation of glomerular filtration rate in children--serum markers and prediction equations].

BACKGROUND: In children, determination of glomerular filtration rate(GFR) is not easy, because serum creatinine (CRE) level changes by their growth and collection of urine is difficult. In this study, we evaluated serum GFR markers and prediction equations of GFR. METHODS: Serum samples were obtained from 200 healthy children; 100 children aged 9 to 10 years and 100 children aged 12 to 13 years (50 males and 50 females respectively). Serum levels of three GFR markers i.e., CRE, cystatin C, and beta2-microglobulin (beta2mG) were measured, and prediction equations of GFR were calculated. Another 110 serum samples were obtained from children aged 0 to 16 years, and the same three markers were measured. RESULTS: In comparing healthy children aged 9 to 10 years and 12 to 13 years, serum CRE levels were significantly higher in the latter group. Serum cystatin C levels were not different between these two groups. Although serum levels of CRE, cystatin C, and beta2mG were not significantly different between males and females aged 9 to 10 years, significantly higher levels of these three markers were observed in males than females aged 12 to 13 years. The reference values of cystatin C and CRE were < or = 0.760 mg/l, < or = 0.623 mg/dl in children aged 9 to 10 years, and < or = 0.835 mg/l, < or = 0.785 mg/dl in children aged 12 to 13 years. Investigation of serum samples obtained from children aged 0 to 16 years suggested the better utility of cystatin C as a GFR marker compared to CRE. The Shwartz prediction equation is considered to be useful among the three CRE-based prediction equations, while the Rule prediction equation is considered to be useful among the three cystatin C-based prediction equations. CONCLUSIONS: We need the reference value of serum CRE in each age and gender to evaluate GFR. And we concluded serum cystatin C is the better GFR marker than CRE in children. As to the prediction equations, the Shwartz and the Rule prediction equation are considered to be practically useful in children.

High-pressure study on bilayer phase behavior of oleoylmyristoyl- and myristoyloleoyl-phosphatidylcholines.

We investigated the thermotropic and barotropic bilayer phase behavior of 1-myristoyl-2-oleoyl-sn-glycero-3-phosphocholine (MOPC) and 1-oleoyl-2-myristoyl-sn-glycero-3-phosphocholine (OMPC) by means of the differential scanning calorimetry (DSC) and high-pressure light-transmittance technique. Water could be used as a solvent for measurements at high pressures because of the elevation of the transition temperatures above 0 degrees C by pressurization, whereas aqueous 50 wt.% ethylene glycol solution was used mainly for those at low pressures. Only one phase transition was observed in the DSC thermogram of the MOPC bilayer membrane as an endothermic peak, and also observed at high pressures as an abrupt change of the light-transmittance. The transition was assigned as a main transition between the lamellar gel (L(beta)) and liquid-crystalline (L(alpha)) phases on the basis of the values of enthalpy change (DeltaH) and slope of the transition temperature with respect to pressure (dT/dP). The DSC thermogram of the OMPC bilayer membrane similarly showed a single endothermic peak but two kinds of phase transitions were observed at different temperatures in the light-transmittance profile at high pressures. The extrapolation of the lower-temperature transition in the high-pressure range to an ambient pressure coincided with the transition observed in the DSC thermogram. This transition was identified as a transition between the lamellar crystal (L(c)) and L(alpha) (or L(beta)) phases from the DeltaH and dT/dP values. The higher-temperature transition, appearing only at high pressures, was identified as the L(beta)/L(alpha) transition considering the topological resemblance of its temperature-pressure phase diagram as that of the dioleoylphosphatidylcholine bilayer membrane. The phase diagram of the OMPC bilayer membrane demonstrated that the L(beta) phase cannot exist at pressures below ca. 190 MPa while it can exist stably in a finite temperature range at pressures above the pressure.

Sources of variation and reference intervals for serum cystatin C in a healthy Japanese adult population.

BACKGROUND: To derive reference intervals (RIs) for cystatin C (CysC), we examined sources of variation in its serum concentration by multiple regression analysis. METHODS: A total of 596 healthy subjects (30-75 years of age) who underwent an annual health check were chosen as candidate reference individuals who did not have any chronic illness requiring medication. Serum CysC was measured together with routine screening tests (basic clinical chemistry and complete blood count). RESULTS: Multivariate analysis revealed that CysC concentrations were higher in males by an average of 0.082 mg/L. Levels were positively associated with age (+0.047 mg/L for every 10 years), body fat (BF%) and cigarette smoking, and negatively associated with alcohol consumption. A subgroup analysis revealed that the gender difference was not significant for those over 50 years of age. RIs were determined using a "latent abnormal values exclusion method" involving deletion of individuals with two or more abnormal results in the screening tests. RIs were partitioned into three categories by age and gender: 0.60-0.95 mg/L for males aged 30-50 years; 0.55-0.84 mg/L for females aged 30-50 years; and 0.64-1.05 mg/L for all subjects aged 51-75 years. Creatinine levels showed little age-related change, but a conspicuous gender difference; they increased with body weight, but not with BF%. CONCLUSIONS: In interpreting serum CysC levels, BF%, cigarette smoking, and alcohol consumption may need to be considered, in addition to age and gender, as sources of variation.

[Evaluation of urinary cystatin C as a marker of renal dysfunction].

BACKGROUND: Urinary excretion of some low molecular weight proteins (LMWPs) is used as an indicator of tubular dysfunction, since they are increased by the damage of tubular reabsorption. Although serum cystatin C is known to be a sensitive marker for GFR, the property of urinary cystatin C as a LMWP has not been fully observed. We evaluated the clinical utility of urinary cystatin C. METHODS: Urine samples were collected from 130 patients with various degree of renal dysfunction, 62 healthy subjects, and 2 patients with acute renal failure, one with renal acute renal failure, the other with prerenal acute renal failure. Urine levels of cystatin C, beta2-microglobulin (beta2mG), and alpha1-microglobulin (alpha1mG) were measured by immunonephelometry. Creatinine clearance(Ccr) tests were conducted on 130 patients with renal dysfunction. Creatinine(CRE) was measured by enzyme assay. RESULTS: The daily urinary excretions of cystatin C and alpha1mG were increased significantly in patients with Ccr<30 ml/min(group I), compared to those in patients with 30 < or = Ccr<70 ml/min(II), and Ccr > or = 70ml/min(III). Although the mean daily excretion of beta2mG increased as Ccr decreased, the significant difference was not observed. The rate of increase in the mean value between III and I was extremely high in cystatin C. Fractional excretions of cystatin C and beta2mG calculated in the same groups increased significantly in I compared to II and III. The rate of increase in the mean value was higher in cystatin C. Regression analyses between urine CRE and each three LMWP gave the best correlation coefficient for cystatin C in healthy subjects. While in one patient with renal acute renal failure, the rate of increase in urine cystatin C was higher than that of other LMWPs, in another patient with prerenal acute renal failure, the rate of increase in urine cystatin C was low. CONCLUSIONS: Although details of urinary movement of LMWPs in nephrons have not been clearly elucidated, the urinary cystatin C seems to have distinctive properties, and to be useful for the evaluation of renal injury.