The Soluble Factor from Oral Cancer Cell Lines Inhibits Interferon-γ Production by OK-432 via the CD40/CD40 Ligand Pathway

OK-432 is a potent immunotherapy agent for several types of cancer, including oral cancer. We previously reported that OK-432 treatment can induce the production of high levels of IFN-γ from peripheral blood mononuclear cells (PBMCs). Moreover, the IFN-γ production from PBMCs by OK-432 is impaired by conditioned media (CM) from oral cancer cells. To determine the inhibitory mechanism of IFN-γ production by CM, the genes involved in IFN-γ production was retrieved by cDNA microarray analysis. We found that CD40 played a key role in IFN-γ production via IL-12 production. Although the expression levels of CD40 were upregulated by OK-432 treatment in PBMCs, CM inhibited OK-432-induced CD40 expression. These findings suggest that uncertain soluble factor(s) in CM may suppress IFN-γ production via the CD40/CD40L–IL-12 axis in PBMCs.(1) Background: OK-432 is a penicillin-killed, lyophilized formulation of a low-toxicity strain (Su) of Streptococcus pyogenes (Group A). It is a potent immunotherapy agent for several types of cancer, including oral cancer. We previously showed that (i) OK-432 treatment induces a high amount of IFN-γ production from peripheral blood mononuclear cells (PBMCs), and (ii) conditioned medium (CM) from oral cancer cells suppresses both the IFN-γ production and cytotoxic activity of PBMCs driven by OK-432. The aim of this study was to determine the inhibitory mechanism of OK-432-induced IFN-γ production from PBMCs by CM. (2) Methods: We performed cDNA microarray analysis, quantitative RT-PCR, and ELISA to reveal the inhibitory mechanism of CM. (3) Results: We found that CD40 plays a key role in IFN-γ production via IL-12 production. Although OK-432 treatment upregulated the expression levels of the IL-12p40, p35, and CD40 genes, CM from oral cancer cells downregulate these genes. The amount of IFN-γ production by OK-432 treatment was decreased by an anti-CD40 neutralizing antibody. (4) Conclusions: Our study suggests that uncertain soluble factor(s) produced from oral cancer cells may inhibit IFN-γ production from PBMCs via suppressing the CD40/CD40L–IL-12 axis

Two-Stage Polyhydroxyalkanoates (PHA) Production from Cheese Whey Using Acetobacter pasteurianus C1 and Bacillus sp. CYR1

Cheese whey (CW) can be an excellent carbon source for polyhydroxyalkanoates (PHA)-producing bacteria. Most studies have used CW, which contains high amounts of lactose, however, there are no reports using raw CW, which has a relatively low amount of lactose. Therefore, in the present study, PHA production was evaluated in a two-stage process using the CW that contains low amounts of lactose. In first stage, the carbon source existing in CW was converted into acetic acid using the bacteria, Acetobacter pasteurianus C1, which was isolated from food waste. In the second stage, acetic acid produced in the first stage was converted into PHA using the bacteria, Bacillus sp. CYR-1. Under the condition of without the pretreatment of CW, acetic acid produced from CW was diluted at different folds and used for the production of PHA. Strain CYR-1 incubated with 10-fold diluted CW containing 5.7 g/L of acetic acid showed the higher PHA production (240.6 mg/L), whereas strain CYR-1 incubated with four-fold diluted CW containing 12.3 g/L of acetic acid showed 126 mg/L of PHA. After removing the excess protein present in CW, PHA production was further enhanced by 3.26 times (411 mg/L) at a four-fold dilution containing 11.3 g/L of acetic acid. Based on Fourier transform infrared spectroscopy (FT-IR), and H-1 and C-13 nuclear magnetic resonance (NMR) analyses, it was confirmed that the PHA produced from the two-stage process is poly-beta-hydroxybutyrate (PHB). All bands appearing in the FT-IR spectrum and the chemical shifts of NMR nearly matched with those of standard PHB. Based on these studies, we concluded that a two-stage process using Acetobacter pasteurianus C1 and Bacillus sp. CYR-1 would be applicable for the production of PHB using CW containing a low amount of lactose

Two-Stage Polyhydroxyalkanoates (PHA) Production from Cheese Whey Using Acetobacter pasteurianus C1 and Bacillus sp. CYR1

Cheese whey (CW) can be an excellent carbon source for polyhydroxyalkanoates (PHA)-producing bacteria. Most studies have used CW, which contains high amounts of lactose, however, there are no reports using raw CW, which has a relatively low amount of lactose. Therefore, in the present study, PHA production was evaluated in a two-stage process using the CW that contains low amounts of lactose. In first stage, the carbon source existing in CW was converted into acetic acid using the bacteria, Acetobacter pasteurianus C1, which was isolated from food waste. In the second stage, acetic acid produced in the first stage was converted into PHA using the bacteria, Bacillus sp. CYR-1. Under the condition of without the pretreatment of CW, acetic acid produced from CW was diluted at different folds and used for the production of PHA. Strain CYR-1 incubated with 10-fold diluted CW containing 5.7 g/L of acetic acid showed the higher PHA production (240.6 mg/L), whereas strain CYR-1 incubated with four-fold diluted CW containing 12.3 g/L of acetic acid showed 126 mg/L of PHA. After removing the excess protein present in CW, PHA production was further enhanced by 3.26 times (411 mg/L) at a four-fold dilution containing 11.3 g/L of acetic acid. Based on Fourier transform infrared spectroscopy (FT-IR), and 1H and 13C nuclear magnetic resonance (NMR) analyses, it was confirmed that the PHA produced from the two-stage process is poly-β-hydroxybutyrate (PHB). All bands appearing in the FT-IR spectrum and the chemical shifts of NMR nearly matched with those of standard PHB. Based on these studies, we concluded that a two-stage process using Acetobacter pasteurianus C1 and Bacillus sp. CYR-1 would be applicable for the production of PHB using CW containing a low amount of lactose

A clinical study on malignant lymphomas arising in the oral region

2008年から2018年までの11年間に当科で口腔領域に発症した悪性リンパ腫の18例について臨床的特徴を調べた。患者は，平均年齢が70.2歳の男性10例，女性8例であった。原発部位の10例（55.6％）は上顎歯肉で，初期症状は15例（83.3％）で腫瘤と腫脹であった。7例（38.9％）が初診時に臨床的に悪性リンパ腫と診断されたが，その他の11例は正しく診断することが困難であった。腫瘤形成や潰瘍のような様々な臨床症状が診断を困難にしており，1回の生検では確定診断が得られなかった。そこで，血液検査値が診断の指標であるかどうかを検討した。われわれの結果は，17例中8例のLDH（47.1％）と16例中11例のsIL-2R（68.8％）が高い値を示していた。さらに，われわれはリンパ球/単球数比（LMR）が診断に役立つかどうかを調べた。LMRの値は，17例中13例（76.5％）が有意に低く，非ML患者と比較しても有意差を認めた。以上のことより，LMRの血液検査も診断の補助的なマーカーであると考えられた。We examined the clinical characteristics of 18 cases of malignant lymphomas arising in the oral region which were diagnosed at our department during the 11 years from 2008 to 2018. The patients consisted of 10 males and 8 females whose mean age was 70.2 years. The primary site was the upper gingiva in 10 cases （55.6％）, and the initial symptoms were tumor mass and swelling in 15 cases （83.3％）. Only 7 cases （38.9％） were clinically diagnosed as malignant lymphoma at first visit, but the other 11 cases were difficult to diagnose correctly. The various clinical symptoms such as mass formation and ulcer made differential diagnosis difficult, and a definitive diagnosis could not be obtained by only a single biopsy. Therefore, we examined whether blood test results might be used as an indicator of the diagnosis or not. Our results showed that LDH in 8 of 17 cases （47.1％） and sIL-2R in 11 of 16 cases （68.8％） were high values. Furthermore, we examined whether lymphocyte-monocyte ratio （LMR） helped the diagnosis or not. Thirteen of 17 cases （76.5％） showed a low value of LMR. There was a significant difference in LMR compared to non-malignant lymphoma patients. LMR in the blood tests was also considered to be a useful marker for the diagnosis