Multivariate Profiling of Metabolites and Volatile Organic Compounds in Citrus depressa Hayata Fruits from Kagoshima, Okinawa, and Taiwan.

Citrus depressa Hayata is a small-fruit citrus species; it is indigenous to Kagoshima, Okinawa, and Taiwan. The metabolites and volatile organic compounds (VOCs) that affect the flavor of its fruits have not been investigated based on geographical origin. In the present study, we investigated the metabolite and VOC profiles of 18 C. depressa cultivation lines from these regions. Multivariate analysis revealed differences in the metabolites of C. depressa based on its cultivation origins; variations in sugar, sugar alcohol, and amino acid contents were also observed. Fruits from Kagoshima and Okinawa had higher galactinol, trehalose, xylose, glucose, and sucrose intensities than fruits from Taiwan (log2-fold change; 2.65-3.44, 1.68-2.13, 1.37-2.01, 1.33-1.57, and 1.07-1.43, respectively), whereas the Taiwanese lines contained higher leucine, isoleucine, serine, and alanine. In contrast to the Taiwanese Nantou line, other cultivation lines had comparable total VOC contents, and the VOCs of all lines were dominated by limonene, γ-terpinene, and p-cymene. Accordingly, the highest VOC intensities were recorded in the Nantou line, which was followed by Shikunin sweet (Kagoshima) and Taoyuan (Taiwan) (log10 normalize concentration; 5.11, 3.08, and 3.01, respectively). Moreover, multivariate analysis plots elucidated the difference in the VOCs of Ishikunibu (Okinawa), Shikunin sweet, and Taoyuan and between those of most Kagoshima and Okinawa cultivation lines. These results suggest that both the cultivation line and origin influence the metabolites and VOCs of C. depressa, thus possibly affecting its flavor quality; the data provide a valuable insight for utilizing C. depressa of different cultivation lines and origins to produce foods and beverages.

A Rapid Method to Measure Serum Retinol Concentrations in Japanese Black Cattle Using Multidimensional Fluorescence.

Vitamin A levels in fattening Japanese Black cattle affect meat quality; therefore, it is important to monitor serum retinol concentrations. To simplify and accelerate the evaluation of serum retinol concentrations in cattle, we developed a new predictive method using excitation-emission matrix (EEM) fluorescence spectrophotometry. For analytical comparison, the concentration of serum retinol was also measured using the conventional HPLC method. We examined excitation (Ex) and emission (Em) wavelengths of cattle serum, which were 250-450 and 250-600 nm, respectively. Parallel factor analysis separated four components from EEM data, one of which was related to retinol. Next, a partial least square regression model was created using the obtained EEMs as explanatory variables and accrual measurement values as objective variables. The determination coefficient value (R2), root mean squared error of prediction (RMSEP), and the ratio of performance to deviation (RPD) of the model were determined. A comparison with reference values found that R2, RMSEP, and RPD of the calibration model were 0.95, 6.4 IU/dl, and 4.2, respectively. This implies that EEM can estimate the serum retinol concentration with high accuracy. Additionally, the fluorescent peaks that contributed to the calibration, which were extracted from the regression coefficient and variable importance in projection plots, were Ex/Em = 320/390 and 330/520 nm. Thus, we assume that this method observes not only free retinol, but also retinol-binding protein. In conclusion, multidimensional fluorescence analysis can accurately and quickly determine serum retinol concentrations in fattening cattle.

Successful continuous nivolumab therapy for metastatic non-small cell lung cancer after local treatment of oligometastatic lesions.

The patient in this report was a 57-year-old man with metastatic non-small cell lung cancer (NSCLC). After no response to two lines of systemic chemotherapy, he was treated with nivolumab as third-line therapy, which resulted in a partial response. After 17 months of nivolumab treatment, he developed bone metastasis in his left femur which was treated with radiation therapy. Nivolumab was restarted after radiation therapy. Four months after radiation therapy, he developed another metastatic lesion in the small intestine which was surgically resected. Because there were no recurrent NSCLC lesions after surgical resection, nivolumab was restarted again. At 18 months after surgery, there were no recurrent NSCLC lesions. Immunohistochemical analysis of peritumoral T lymphocytes showed higher expression of T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) and lymphocyte activation gene 3 (LAG-3) in recurrent lesions of bone and small intestine than in primary lesions. Upregulation of TIM-3 and LAG-3 could be associated with mechanisms of adaptive resistance to nivolumab in this case. Here, we report a successful case of continued nivolumab therapy with remission after local treatments consisting of radiation therapy and surgical resection for oligometastases. Continuation of immune checkpoint inhibitor (ICI) treatment may be worth considering if oligometastases can be controlled. KEY POINTS: Significant findings of the study We report a successful case of continued nivolumab treatment with remission after local treatment (radiation therapy and surgical resection) for oligometastases. What this study adds Upregulation of T cell immunoglobulin and mucin domain-containing protein 3 and lymphocyte-activation gene 3 could be associated with mechanisms of adaptive resistance to nivolumab.

Metabolomic Evaluation of the Quality of Leaf Lettuce Grown in Practical Plant Factory to Capture Metabolite Signature.

Vegetables produce metabolites that affect their taste and nutritional value and compounds that contribute to human health. The quality of vegetables grown in plant factories under hydroponic cultivation, e.g., their sweetness and softness, can be improved by controlling growth factors including the temperature, humidity, light source, and fertilizer. However, soil is cheaper than hydroponic cultivation and the visual phenotype of vegetables grown under the two conditions is different. As it is not clear whether their metabolite composition is also different, we studied leaf lettuce raised under the hydroponic condition in practical plant factory and strictly controlled soil condition. We chose two representative cultivars, "black rose" (BR) and "red fire" (RF) because they are of high economic value. Metabolite profiling by comprehensive gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) resulted in the annotation of 101 metabolites from 223 peaks detected by GC-MS; LC-MS yielded 95 peaks. The principal component analysis (PCA) scatter plot showed that the most distinct separation patterns on the first principal component (PC1) coincided with differences in the cultivation methods. There were no clear separations related to cultivar differences in the plot. PC1 loading revealed the discriminant metabolites for each cultivation method. The level of amino acids such as lysine, phenylalanine, tryptophan, and valine was significantly increased in hydroponically grown leaf lettuce, while soil-cultivation derived leaf lettuce samples contained significantly higher levels of fatty-acid derived alcohols (tetracosanol and hexacosanol) and lettuce-specific sesquiterpene lactones (lactucopicrin-15-oxalate and 15-deoxylactucin-8-sulfate). These findings suggest that the metabolite composition of leaf lettuce is primarily affected by its cultivation condition. As the discriminant metabolites reveal important factors that contribute to the nutritional value and taste characteristics of leaf lettuce, we performed comprehensive metabolite profiling to identify metabolite compositions, i.e., metabolite signature, that directly improve its quality and value.

[A case of severe COPD associated with tracheo-bronchial stenosis, treated with non-invasive positive pressure ventilation].

A 76-year-old man was admitted with acute exacerbation of COPD. He was administered bronchodilators, antibiotics and oral corticosteroids. Although his cough, sputum, fever and the laboratory data improved. wheezing and dyspnea remained. The chest CT revealed severe stenosis of the trachea and both main bronchi, which was thought to be the cause of the symptoms, and similar to the condition of "excessive dynamic airway collapse (EDAC)". We treated him with NPPV and his symptoms improved and he returned home. "EDAC"-like tracheobronchial stenosis with COPD treated with NPPV is rare.

[Direct hemoperfusion with a polymyxin B-immobilized fiber column eliminates neutrophils and improves pulmonary oxygenation--a comparison of two cases with acute exacerbation of idiopathic pulmonary fibrosis].

Idiopathic pulmonary fibrosis (IPF) is characterized by radiographically evident interstitial infiltrates predominantly affecting the lung bases and by progressive dyspnea and worsening pulmonary function. Acute exacerbation of IPF is recognized widely as an accelerated phase occurring suddenly in the course of IPF, which leads to a catastrophic outcome. No treatment has proven to be effective so far. We describe two cases of acute exacerbation of IPF which were treated by direct hemoperfusion with a polymyxin B-immobilized fiber column (PMX) after steroid pulse therapy. One patient died on the 21st day after direct hemoperfusion and the other is alive at present. The survivor showed significant improvement of pulmonary oxygenation correlating with a decrease in the neutrophil count, SP-D and KL-6 after direct hemoperfusion with PMX. On the other hand, the non-survivor showed no improvement of pulmonary oxygenation, SP-D and KL-6 despite a decrease in neutrophil count comparable with that of the survivor. Neutrophil count which decreased temporarily after direct hemoperfusion with PMX soon convert to increase in the non-survivor, which is a characteristic difference between two. We reached the conclusions that (1) direct hemoperfusion with PMX absorbs neutrophils and this mechanism is effective to improve pulmonary oxygenation, (2) sometimes neutrophil absorption is not enough to control pulmonary inflammation in patients with acute exacerbation of IPF.

Propionibacterium acnes-induced hepatic granuloma formation is impaired in mice lacking tetraspanin CD9.

The granuloma is a host defence response to persistent pathogenic irritants. In the process of granuloma formation, the activation, migration, and fusion of macrophages occur locally, but the mechanisms involved remain elusive. Tetraspanins regulate cell migration and fusion by organizing functional molecular complexes in membrane microdomains. Here we investigated the role of tetraspanin CD9 in hepatic granuloma formation. Immunostaining of the liver of untreated wild-type mice showed that CD9 was expressed by vascular endothelial cells and perivenular hepatocytes. When intrahepatic granulomas were induced by intravenous injection of Propionibacterium acnes, hepatocyte CD9 was extensively upregulated, while inflammatory cells constituting granulomas were mostly negative for CD9. Compared with wild-type littermates, CD9-knockout mice showed dissemination of Propionibacterium acnes and reduced number and size of granulomas after the injection. Moreover, production of granuloma-inducing cytokines, TNF-alpha and IFN-gamma, was delayed and chemotactic activity for macrophages was suppressed in the liver of mutant mice. These results suggest that CD9 is one of the proteins that promotes granuloma formation in the liver.