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Background: Histamine fixed-immunoglobulin formulations, which consisted of 0.15 µg of histamine dihydrochloride and 12 mg of human immunoglobulin in a vial, are used for anti-allergic treatments, and controlling the amounts of histamine in the formulations is essential to avoid histamine intoxication., Objective: A high-performance liquid chromatography (HPLC) method for determination of histamine contents of the formulations was established and validated., Methods: Histamine extracted from the formulation was labeled with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate and was analyzed by gradient elution HPLC with UV detection at 260 nm., Results: The method showed linearity in the range 0.8-2.4 µM (R > 0.999), accuracy (100.1-105.8% recovery), and precision (relative standard deviation ≤ 1.93%). The validated method was applied for five lots of the pharmaceutical, and their histamine contents were determined to be 0.149-0.155 µg/vial., Conclusions: These results indicated that the validated method is useful to control amounts of histamine in biopharmaceutical products., Highlights: The HPLC method was developed for quantitative determination of histamine content of the histamine fixed-immunoglobulin formulations., (© AOAC INTERNATIONAL 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Purpose: To evaluate the detailed immunosuppressive role(s) of PD-L2 given that its detailed role(s) remains unclear in PD-1 signal blockade therapy in animal models and humans., Experimental Design: We generated mouse cell lines harboring various status of PD-L1/PD-L2 and evaluated the tumor growth and phenotypes of tumor-infiltrated lymphocytes using several PD-1 signal blockades in animal models. In humans, the correlation between immune-related gene expression and CD274 (encoding PD-L1) or PDCD1LG2 (encoding PD-L2) was investigated using The Cancer Genome Atlas (TCGA) datasets. In addition, PD-L1 or PD-L2 expression in tumor cells and CD8 + T-cell infiltration were assessed by IHC., Results: In animal models, we showed that PD-L2 expression alone or simultaneously expressed with PD-L1 in tumor cells significantly suppressed antitumor immune responses, such as tumor antigen-specific CD8 + T cells, and was involved in the resistance to treatment with anti-PD-L1 mAb alone. This resistance was overcome by anti-PD-1 mAb or combined treatment with anti-PD-L2 mAb. In clinical settings, antitumor immune responses were significantly correlated with PD-L2 expression in the tumor microenvironment in renal cell carcinoma (RCC) and lung squamous cell carcinoma (LUSC)., Conclusions: We propose that PD-L2 as well as PD-L1 play important roles in evading antitumor immunity, suggesting that PD-1/PD-L2 blockade must be considered for optimal immunotherapy in PD-L2-expressing cancers, such as RCC and LUSC., (©2019 American Association for Cancer Research.)

A mammalian body is composed of more than 200 different types of cells. The purification of a certain cell type from tissues/organs enables a wide variety of studies. One popular cell purification method is immunological isolation, using antibodies against specific cell surface antigens. However, this is not a general-purpose method, since suitable antigens have not been found in certain cell types, including embryonic gonadal somatic cells and Sertoli cells. To address this issue, we established a knock-in mouse line, named R26 KI, designed to express the human cell surface antigen hCD271 through Cre/loxP-mediated recombination. First, we used the R26 Kl mouse line to purify embryonic gonadal somatic cells. Gonadal somatic cells were purified from the R26 KI; Nr5a1-Cre-transgenic (tg) embryos almost equally as efficiently as from Nr5a1-hCD271-tg embryos. Second, we used the R26 KI mouse line to purify Sertoli cells successfully from R26 KI; Amh-Cre-tg testes. In summary, we propose that the R26 KI mouse line is a powerful tool for the purification of various cell types., (© 2015 Wiley Periodicals, Inc.)

Background: Recently, it has been reported that Th17 contributes to allograft rejection after transplantation. We investigated the alteration of Th17 and regulatory T cells (Treg) distribution in an animal model of bronchiolitis obliterans following ectopic tracheal transplantation model., Methods: Tracheal grafts from B6 mice transplanted into subcutaneous sites of C3H mice. Allografts were histologically evaluated, and expressions of CD4, CD8, CD25, CD28, CD127, CD152 and Foxp3, and intracellular interleukin (IL)-4, -6, -17, and interferon-gamma, in peripheral blood lymphocytes were analyzed. Tracheal graft IL-6 and -17 mRNA expression was assessed using a quantitative reverse-transcriptase polymerase chain reaction. All the data in allogenic transplantation was compared with those in isograft controls. In addition, the effect of IL-6 neutralization on the allograft was evaluated with histopathology and the IL-17 mRNA expression., Results: Treg was significantly lower in peripheral blood of allogenic mice, whereas no significant difference in Th17 in the CD4 T-cell population was observed after allogenic or isogenic transplantation. Locoregional histologic examination revealed the presence of IL-6-producing lymphocytes and endothelium in the allograft, and the luminal obliteration by fibroblast proliferation. Both IL-6 and IL-17 mRNA levels were elevated in the allograft. Severity of tracheal obliteration and IL-17 mRNA level was significantly suppressed in the IL-6 neutralized allografts., Conclusions: After allograft in a mouse bronchiolitis obliterans model, IL-17 production increases locally without an alteration in peripheral blood Th17 cells, whereas peripheral Tregs decreases. Th17 cells, which can be regulated by IL-6 stimulation, may play a role in posttransplantation rejection of the allograft.

Parkinson's disease (PD) is a common neurodegenerative disorder. The motor neuron degeneration 2 mutant (mnd2) mouse exhibits loss of striatal neurons, muscle wasting, weight loss, and death within 40days of birth, and is considered to be a useful animal model of PD. mnd2 was identified as an autosomal recessive mutation in the HtrA2/Omi gene, which encodes a mitochondrial serine protease. Omi-deficient mitochondria are more sensitive to mitochondrial permeability transition (mPT), which raises the possibility that mPT plays a role in motor neurodegeneration in mnd2 mice. Given that cyclophilin D (CypD)-deficient mitochondria are resistant to mPT, we examined whether CypD-dependent mPT is involved in the pathogenesis of neurodegenerative disorders in mnd2 mice by generating CypD-deficient mnd2 mice. Brain mitochondria isolated from CypD-deficient mnd2 mice were more resistant to Ca(2+)-induced mPT than those of mnd2 mice. However, both mnd2 mice and CypD-deficient mnd2 mice showed similar survival periods and phenotypes, including the lack of weight gain, muscle wasting, and resting tremor. Our data suggest that CypD-dependent mPT does not play a major role in neurodegeneration in mnd2 mice., (2010 Elsevier Inc. All rights reserved.)

A lobectomy is the standard surgical procedure for non-small cell lung cancer (NSCLC), though recently a limited resection is more likely chosen for small-sized early stage disease. To elucidate the effectiveness of an intentional segmentectomy as a curative procedure, factors associated with survival after the procedure were examined in a long-term retrospective study carried out at a single institute. Patients with stage I, II, or III disease NSCLC who underwent a segmentectomy between 1980 and 2002 (n = 144) were retrospectively studied and the results compared with those who underwent a lobectomy during the same period (n = 1241). Tumor size, nodal involvement, pleural involvement, and histological type were independent significant prognostic factors in patients who received a segmentectomy. Six patients had a large cell carcinoma and each died from the disease within 5 years after the segmentectomy. In patients with p-T1N0M0 (stage IA) disease and a tumor smaller than 2 cm, except for large cell carcinomas, the 5- and 10-year survival rates were 83% and 83%, respectively, after a segmentectomy, and 81% and 64%, respectively, after a lobectomy (p = 0.66). In patients with p-T1N0M0 disease and a tumor diameter exceeding 2 cm, the 5- and 10-year survival rates were 58% and 58%, respectively, after a segmentectomy, and 78% and 60%, respectively, after a lobectomy (p = 0.057). We concluded that histological type and tumor size were relevant for determining the indication of an intentional segmentectomy for NSCLC with stage IA disease.

We report a 64-year-old woman treated with surgical intervention for late-onset chylothorax following a pleuropneumonectomy. The patient underwent an extrapleural pneumonectomy for diffuse malignant mesothelioma and was uneventfully discharged on postoperative day 29. Pleural effusion aspirated on postoperative day 9 was dark red. A chest roentgenogram taken at our outpatient clinic revealed a mediastinal shift on postoperative day 56. No bacterial infection was found in the milky effusion. We made a diagnosis of postoperative late-onset chylothorax based on the laboratory data obtained from tests of the pleural fluid. A repeat thoracotomy to ligate the lymphatic duct was performed because conservative management with chest tube drainage and no oral feeding was unsuccessful. The patient was discharged after the operation with a good clinical course.

Objective: The number of surgical interventions for small-sized lung cancer has increased with the development of computed tomography. We attempted to identify clinicopathologic characteristics of peripheral, small-sized, non-small cell lung cancer to show the limitation of partial resection or segmentectomy., Methods: A retrospective analysis of 143 patients who underwent a complete resection for a peripheral non-small cell lung cancer of 2 cm or less in diameter was performed. The relationships between nodal involvement and other clinical factors were also assessed in patients who underwent a lobectomy plus node dissection., Results: The overall 5-year survival rate was 88.1%. The 5-year survival rate was 100% for patients with a tumor of 1.5 cm or less. Survival for patients with adenocarcinoma histology was significantly better than for those with nonadenocarcinoma histology (P = .03). The 5-year survival rate for patients without lymph node metastases was 91.6%, whereas it was 62.5% for those with nodal involvement (P < .01). Increase of prethoracotomy serum carcinoembryonic antigen level was an independent predictor of a poor prognosis. Lymph node metastasis was significantly increased in those with pleural invasion by the primary lesion and increased serum carcinoembryonic antigen level. Fourteen (16.9%) of 83 patients with a tumor diameter of larger than 1.5 cm had nodal metastasis., Conclusions: Nodal involvement should be considered in patients with non-small cell lung cancer of 2 cm or less in diameter who show pleural invasion or an increased carcinoembryonic antigen level. A lobectomy with node dissection is recommended for patients with a tumor larger than 1.5 cm, suspected pleural invasion, or prethoracotomy carcinoembryonic antigen level increase.

Background: A pulmonary metastasectomy for colorectal carcinoma is a generally accepted procedure, although several prognostic predictors have been reported. The aim of this study is to define the patients who benefit from pulmonary metastasectomy for colorectal carcinoma., Methods: Retrospective survival analysis was performed using 128 patients who underwent curative pulmonary resection., Results: The overall 5-year survival rate was 45.3%. Univariate analysis showed the number of metastases, location (unilateral or bilateral), prethoracotomy carcinoembryonic antigen (CEA) level, hilar or mediastinal lymph-node metastasis, and Dukes' stage to be considerable prognostic factors. Among these, Dukes' A for the primary lesion and unilateral pulmonary metastasis were shown to be independent predictors of longer survival by multivariate analysis (p = 0.0093 and p = 0.0182, respectively). In patients treated with both pulmonary and hepatic metastastasectomies, a better prognosis was observed in those who received metachronous resection. Recurrence after a pulmonary metastasectomy frequently occurred in the thorax and the 3-year survival rate was 44.6% in patients who underwent a repeat thoracotomy., Conclusions: Patients with unilateral metastasis and Dukes' A for the primary tumor benefit most from the resection of pulmonary metastasis from colorectal carcinoma. Further, the number of metastases, prethoracotomy CEA level, and hilar or mediastinal lymph-node involvement should be considered to determine the operative indication. Finally, periodic follow-up examinations for thoracic recurrence should be carefully performed as these patients may have a heightened risk of requiring a repeat thoracotomy.

The results of treatment of gastric cancer with docetaxel have scarcely been reported. Combination chemotherapy of docetaxel and doxifluridine was administered to five patients with recurrent or unresectable gastric cancer in the authors' department, and the overall response rate was 40%. Among them, we report here a case of multiple skin metastases successfully treated with this chemotherapy. A 71-year-old man had already undergone total gastrectomy with splenectomy about one year previously because of advanced gastric cancer: type 3, tub2, se, n1, INF gamma, ly3(+), v1(+), stage IIIa. He was treated with adjuvant chemotherapy of low-dose FP (CDDP/5-FU) for two weeks, and thereafter 5-FU (200 mg/day orally) was administered. Skin metastases appeared on the right upper eyelid and the left femoral region, though no recurrent findings were detected by CT, US, and the like one year after operation. The combination chemotherapy of docetaxel and doxifluridine was administered because multiple skin tumors rapidly appeared on the back and the right upper eyelid region. These tumors disappeared after 3 weeks and he has had no recurrence more than one year after chemotherapy.

Pancreatic islet cell tumors are rarely associated with intra-abdominal hemorrhage. We report herein a rare case of nonfunctioning islet cell carcinoma associated with massive hemorrhage into the abdominal cavity caused by spontaneous rupture of the tumor. A 44-year-old man presenting with sudden upper abdominal pain was admitted to his local hospital on April 18, 1994. On April 19, a laparotomy was performed with the diagnosis of peritonitis. Massive hemorrhage of unknown origin occurred, and he was transferred to our hospital in a state of hypovolemic shock. Imaging findings revealed massive hematoma in the abdominal cavity and a hypervascular tumor arising from the body of the pancreas. Because the hemorrhage was life-threatening, an emergent re-laparotomy was performed on April 20. Apart from the massive hemorrhage, a pancreatic tumor (60 x 35 x 30 mm in size) with spontaneous rupture was noted. Distal pancreatectomy, combined with splenectomy and removal of the hematoma, was performed. Histological findings revealed an islet cell carcinoma of the pancreas with venous invasion. Peritoneal dissemination, liver metastasis, and lymph node metastasis were not observed. The patient is alive without recurrence 6 years and 5 months after the operation.

Adrenergic regulation of prostaglandin (PG) biosynthesis was investigated in 1-14C-arachidonic acid-prelabeled cultured rabbit gastric epithelial cells (RGECs). RGECs expressed adrenergic alpha 1- and beta-receptors and muscarinic receptors. Norepinephrine facilitated the synthesis of PGs I2 (determined as the stable metabolite 6-keto PGF1 alpha) and E2 and hydroxyfatty acids, while epinephrine facilitated the synthesis of PGI2 and hydroxyfatty acids, but not PGE2. However, isoproterenol did not affect PG biosynthesis. The effects of norepinephrine and epinephrine on PG biosynthesis were markedly suppressed by the non-selective alpha-blocker phentolamine and/or the selective alpha 1-blocker prazosin. In combination with epinephrine, the selective alpha 2-blocker yohimbine or the beta-blocker propranolol facilitated PGE2 synthesis. Acetylcholine did not affect PG biosynthesis. These results indicate that norepinephrine and epinephrine act on PG biosynthesis as adrenergic agonists in these cultured RGECs and that an alpha 1-receptor couples PGI2 and PGE2 synthesis.

1. After a single oral dose of d-limonene (200-1200 mg/kg) no effects were observed on liver triglyceride, microsomal protein, cytochrome b5, and the drug-metabolizing enzymes. Glycogen content was slightly decreased at doses higher than 800 mg/kg, and cytochrome P-450 and delta-aminolaevulinic acid synthetase activity was slightly increased at 1200 mg/kg. 2. After repeated treatment (400 mg/kg/day) for 30 days, the relative liver weight and hepatic phospholipid content were only slightly increased, and liver and serum cholesterol were decreased 49 and 8%, respectively. Of the phospholipid fatty acids, palmitic, linoleic and arachidonic acids were increased, and stearic acid was decreased. Aminopyrine demethylase and aniline hydroxylase were increased 26 and 22%, respectively, and cytochrome P-450 and b5 were likewise increased 31 and 30%.