Enriching CCL3 in the Tumor Microenvironment Facilitates T cell Responses and Improves the Efficacy of Anti-PD-1 Therapy

Chemokines are key factors that influence the migration and maintenance of relevant immune cells into an infected tissue or a tumor microenvironment. Therefore, it is believed that the controlled administration of chemokines in the tumor microenvironment may be an effective immunotherapy against cancer. Previous studies have shown that CCL3, also known as macrophage inflammatory protein 1-alpha, facilitates the recruitment of dendritic cells (DCs) for the presentation of tumor Ags and promotes T cell activation. Here, we investigated the role of CCL3 in regulating the tumor microenvironment using a syngeneic mouse tumor model. We observed that MC38 tumors overexpressing CCL3 (CCL3-OE) showed rapid regression compared with the wild type MC38 tumors. Additionally, these CCL3-OE tumors showed an increase in the proliferative and functional tumor-infiltrating T cells. Furthermore, PD-1 immune checkpoint blockade accelerated tumor regression in the CCL3-OE tumor microenvironment. Next, we generated a modified CCL3 protein for pre-clinical use by fusing recombinant CCL3 (rCCL3) with a non-cytolytic hybrid Fc (HyFc).Administering a controlled dose of rCCL3-HyFc via subcutaneous injections near tumors was effective in tumor regression and improved survival along with activated myeloid cells and augmented T cell responses. Furthermore, combination therapy of rCCL3-HyFc with PD-1 blockade exhibited prominent effect to tumor regression. Collectively, our findings demonstrate that appropriate concentrations of CCL3 in the tumor microenvironment would be an effective adjuvant to promote anti-tumor immune responses, and suggest that administering a long-lasting form of CCL3 in combination with PD-1 blockers can have clinical applications in cancer immunotherapy.

Enriching CCL3 in the Tumor Microenvironment Facilitates T cell Responses and Improves the Efficacy of Anti-PD-1 Therapy

Chemokines are key factors that influence the migration and maintenance of relevant immune cells into an infected tissue or a tumor microenvironment. Therefore, it is believed that the controlled administration of chemokines in the tumor microenvironment may be an effective immunotherapy against cancer. Previous studies have shown that CCL3, also known as macrophage inflammatory protein 1-alpha, facilitates the recruitment of dendritic cells (DCs) for the presentation of tumor Ags and promotes T cell activation. Here, we investigated the role of CCL3 in regulating the tumor microenvironment using a syngeneic mouse tumor model. We observed that MC38 tumors overexpressing CCL3 (CCL3-OE) showed rapid regression compared with the wild type MC38 tumors. Additionally, these CCL3-OE tumors showed an increase in the proliferative and functional tumor-infiltrating T cells. Furthermore, PD-1 immune checkpoint blockade accelerated tumor regression in the CCL3-OE tumor microenvironment. Next, we generated a modified CCL3 protein for pre-clinical use by fusing recombinant CCL3 (rCCL3) with a non-cytolytic hybrid Fc (HyFc).Administering a controlled dose of rCCL3-HyFc via subcutaneous injections near tumors was effective in tumor regression and improved survival along with activated myeloid cells and augmented T cell responses. Furthermore, combination therapy of rCCL3-HyFc with PD-1 blockade exhibited prominent effect to tumor regression. Collectively, our findings demonstrate that appropriate concentrations of CCL3 in the tumor microenvironment would be an effective adjuvant to promote anti-tumor immune responses, and suggest that administering a long-lasting form of CCL3 in combination with PD-1 blockers can have clinical applications in cancer immunotherapy.

Protein kinase C beta II upregulates intercellular adhesion molecule-1 via mitochondrial activation in cultured endothelial cells

Activation of protein kinase C (PKC) is closely linked with endothelial dysfunction. However, the effect of PKCβII on endothelial dysfunction has not been characterized in cultured endothelial cells. Here, using adenoviral PKCβII gene transfer and pharmacological inhibitors, the role of PKCβII on endothelial dysfucntion was investigated in cultured endothelial cells. Phorbol 12-myristate 13-acetate (PMA) increased reactive oxygen species (ROS), p66shc phosphorylation, intracellular adhesion molecule-1, and monocyte adhesion, which were inhibited by PKCβi (10 nM), a selective inhibitor of PKCβII. PMA increased the phosphorylation of CREB and manganese superoxide dismutase (MnSOD), which were also inhibited by PKCβi. Gene silencing of CREB inhibited PMA-induced MnSOD expression, suggesting that CREB plays a key role in MnSOD expression. Gene silencing of PKCβII inhibited PMA-induced mitochondrial ROS, MnSOD, and ICAM-1 expression. In contrast, overexpression of PKCβII using adenoviral PKCβII increased mitochondrial ROS, MnSOD, ICAM-1, and p66shc phosphorylation in cultured endothelial cells. Finally, PKCβII-induced ICAM-1 expression was inhibited by Mito-TEMPO, a mitochondrial ROS scavenger, suggesting the involvement of mitochondrial ROS in PKC-induced vascular inflammation. Taken together, the results suggest that PKCβII plays an important role in PMA-induced endothelial dysfunction, and that the inhibition of PKCβII-dependent p66shc signaling acts as a therapeutic target for vascular inflammatory diseases.

Trichostatin A Modulates Angiotensin II-induced Vasoconstriction and Blood Pressure Via Inhibition of p66shc Activation

Histone deacetylase (HDAC) has been recognized as a potentially useful therapeutic target for cardiovascular disorders. However, the effect of the HDAC inhibitor, trichostatin A (TSA), on vasoreactivity and hypertension remains unknown. We performed aortic coarctation at the inter-renal level in rats in order to create a hypertensive rat model. Hypertension induced by abdominal aortic coarctation was significantly suppressed by chronic treatment with TSA (0.5 mg/kg/day for 7 days). Nicotinamide adenine dinucleotide phosphate-driven reactive oxygen species production was also reduced in the aortas of TSA-treated aortic coarctation rats. The vasoconstriction induced by angiotensin II (Ang II, 100 nM) was inhibited by TSA in both endothelium-intact and endothelium-denuded rat aortas, suggesting that TSA has mainly acted in vascular smooth muscle cells (VSMCs). In cultured rat aortic VSMCs, Ang II increased p66shc phosphorylation, which was inhibited by the Ang II receptor type I (AT1R) inhibitor, valsartan (10 microM), but not by the AT2R inhibitor, PD123319. TSA (1~10 microM) inhibited Ang II-induced p66shc phosphorylation in VSMCs and in HEK293T cells expressing AT1R. Taken together, these results suggest that TSA treatment inhibited vasoconstriction and hypertension via inhibition of Ang II-induced phosphorylation of p66shc through AT1R.

Redox Regulating Protein APE1/Ref-1 Expression is Increased in Abdominal Aortic Coarctation-induced Hypertension Rats

BACKGROUND: Aim of study is designed to investigate whether apurinic/apyrimidinic endonuclease-1/redox factor-1 (APE1/Ref-1) expression is changed in abdominal aortic coarctation models. METHODS: Male Sprague-Dawley rats were randomly assigned with abdominal aortic coarctation, repaired group, sham, and control groups. Endothelial function was assessed with endothelium-dependent relaxations. Detection of superoxide anion and lipid peroxidation was performed by lucigenin chemiluminescence and thiobarbituric acid-reactive substances assay. APE1/Ref-1 expression was measured with Western blot and immunohistochemistry. RESULTS: In anesthetized condition, the abdominal aortic coarctation rats showed hypertension as systolic/diastolic arterial pressure of 171/114 mm Hg, compared with 114/94 mm Hg of control. Endothelium-dependent relaxations were significantly impaired in the aortic coarctation which was recovered in 1 week after coarctation repair. Superoxide production and lipid peroxidation were elevated in aortic coarctation rats. In immunohistochemistry, APE1/Ref-1 expressions were increased at aorta and kidney in aortic coarctation rats. Increased APE1/Ref-1 expression in aorta was recovered by repair of coarctation. CONCLUSIONS: Taken together, it suggests that APE1/Ref-1 expression was increased in aortic coarctation-induced hypertensive rats, suggesting a biomarker for hypertension. Impaired endothelium dependent relaxation in the aortic coarctation can be modulated by repair of coarctation or the modulation of blood pressure.

Cerebral oxygenation monitoring of patients during arthroscopic shoulder surgery in the sitting position / 대한마취과학회지

BACKGROUND: Cerebral hypotension and desaturation can occur during shoulder surgery in the seated position. We evaluated the correlation of cerebral oxygen saturation (rSO2) using near infra-red spectroscopy (NIRS) and mean arterial pressures (MAP) (at the levels of the brain and heart). METHODS: Fifty patients, scheduled for the arthroscopic shoulder surgery in the seated position, were enrolled to monitor the rSO2, bispectral Index (BIS), and MAPs at the levels of the brain and heart. The values of each parameter were collected at 5 min after intubation, immediately after placing the patient in the sitting position, 5 min after the patient was seated, immediately after the surgical incision, and every 30 min after incision. RESULTS: A correlation between the cerebral rSO2 and the MAP at the level of brain were statistically significant. Cerebral rSO2 and MAP after a change of posture from supine to sitting position were significantly decreased, compared to the baseline value. CONCLUSIONS: Monitoring cerebral rSO2 and MAP at the level of brain can be helpful to detect the possibility of cerebral deoxygenation earlier.

Vasorelaxing Activity of Ulmus davidiana Ethanol Extracts in Rats: Activation of Endothelial Nitric Oxide Synthase

Ulmus davidiana var. japonica Rehder (Urticales: Ulmaceae) (UD) is a tree widespread in northeast Asia. It is traditionally used for anticancer and anti-inflammatory therapy. The present study investigated the effect of an ethanol extract of UD on vascular tension and its underlying mechanism in rats. The dried root bark of UD was ground and extracted with 80% ethanol. The prepared UD extract was used in further analysis. The effect of UD on the cell viability, vasoreactivity and hemodynamics were investigated using propidium iodide staining in cultured cells, isometric tension recording and blood pressure analysis, respectively. Low dose of UD (10~100microg/ml) did not affect endothelial cell viability, but high dose of UD reduced cell viability. UD induced vasorelaxation in the range of 0.1~10microg/ml with an ED50 value of 2microg/ml. UD-induced vasorelaxation was completely abolished by removal of the endothelium or by pre-treatment with L-NAME, an inhibitor of nitric oxide synthase. UD inhibited calcium influx induced by phenylephrine and high K+ and also completely abolished the effect of L-NAME. Intravenous injection of UD extracts (10~100 mg/kg) decreased arterial and ventricular pressure in a dose-dependent manner. Moreover, UD extracts reduced the ventricular contractility (+dP/dt) in anesthetized rats. However, UD-induced hypotensive actions were minimized in L-NAME-treated rats. Taken together, out results showed that UD induced vasorelaxation and has antihypertensive properties, which may be due the activation of nitric oxide synthase in endothelium.

Endocrine Disruptors

For the previous century, the humans have created an unintended and unwanted problem of endocrine disruptors as a potential threat to our public health. By the name of industrialization, endocrine disruptors are smuggling in the everyday life of people today. Although there are much debate on the reality of their emerging health threat, it is no doubt that there are certain classes of compounds that have the potential to affect hormonal status adversely, leading to abnormal development, reproductive dysfunction, and some cancers. The classes of endocrine disruptors are extensively diverse and even more increasing, such as, polychlorinated biphenyls (PCBs), dioxins, dieldrin, bisphenol A and toxaphene. Although these endocrine disruptors have been prohibited or tightly regulated, many of them are still unrecognized and still used without knowing their potential threat to the biological world. Once they are released into the environment, they usually persist without degradation and even undergo bioaccumulation and bioconcentration in food chain. Comparing with the great concern over the public health, we do not have enough information for these issues. It is now clear that we need further extensive studies for the risk assessment and the protection of human and ecological health from the potential hazards of endocrine disruptors. This article introduces a breif overview of the current status of our knowledge and research on endocrine disruptors.

NADPH Oxidase and Mitochondrial ROS are Involved in the TNF-alpha-induced Vascular Cell Adhesion Molecule-1 and Monocyte Adhesion in Cultured Endothelial Cells

Atherosclerosis is considered as a chronic inflammatory process. However, the nature of the oxidant signaling that regulates monocyte adhesion and its underlying mechanism is poorly understood. We investigated the role of reactive oxygen species on the vascular cell adhesion molecule-1 (VCAM-1) and monocyte adhesion in the cultured endothelial cells. TNF-alpha at a range of 1~30 ng/ml induced VCAM-1 expression dose-dependently. BCECF-AM-labeled U937 cells firmly adhered on the surface of endothelial cells when the endothelial cells were incubated with TNF-alpha (15 ng/ml). Ten micromol/L of SB203580, an inhibitor of p38 MAPK, significantly reduced TNF-alpha-induced VCAM-1 expression, compared to the JNK inhibitor (40micromol/L of SP60015) or ERK inhibitor (40micrommol/L of U0126). Also, SB203580 significantly inhibited TNF-alpha-induced monocyte adhesion in HUVEC. Superoxide production was minimal in the basal condition, however, treatment of TNF-alpha induced superoxide production in the dihydroethidine-loaded endothelial cells. Diphenyleneiodonium (DPI, 10micromol/L), an inhibitor of NADPH oxidase, and rotenone (1micromol/L), an inhibitor of mitochondrial complex I inhibited TNF-alpha-induced superoxide production, VCAM-1 expression and monocyte adhesion in the endothelial cells. Taken together, our data suggest that NADPH oxidase and mitochondrial ROS were involved in TNF-alpha-induced VCAM-1 and monocyte adhesion in the endothelial cells.

Distribution and Correlates of Obesity in a Korean Rural People / 한국역학회지

OBJECTIVE: The aim of present study was to assess the distribution and correlates of obesity in a Korean rural people using both body mass index(BMI) and body fat percent. METHODS: A total of 1,243 participants were recruited using a two-staged stratified sampling. A structured questionnaire was used to ask their sociodemographics (gender, age, marital status, educational background, and etc.) and health-related behaviors (smoking, drinking, and regular exercise etc.). The data of weight-for height, and body fat percent were also collected by physical examination. For the estimation and analysis of correlates of obesity, we used BMI(>or=25kg/m2) and body fat percent(male>or=25%, female>or=30) as a cut-point of obesity. All analyses were stratified to three age groups(>20, 20-39, 40>or=). RESULTS: The prevalence of obesity in this study was higher than that in previous studies. This study showed that 32.6% (male: 33.7%, female: 31.7%) of participants according to BMI, and 45.6% (male: 43.8%, female: 47.4%) of them according to body fat percent were obese group. Logistic regression analysis showed that, in the criteria of BMI, while obesity was associated with female and low educational background under the age of 40, prevalence of obesity was higher in non smokers over the age of 40. In case of body fat percent crteria, single(unmarried, divorced, separated, widowed) were more likely to be obese in male aged 20 to 39 years. In the group aged 40 years and over, risk of obesity was higher in female than in male. CONCLUSION: This result suggests that obesity is common in Korean rural areas, especially among adolescents and female aged 40 years or over, and the risk factors for obesity were different by age. It is required that health management program focusing on obesity and its adverse outcomes should be developed in a community setting.

Smoking Status and Smoking Cessation Activity among Physicians in a Community / 예방의학회지

OBJECTIVES: The purposes of this study were to assess the smoking status, knowledge and attitude related to smoking and smoking cessation activity of the physicians in a community, and to identify their predictors of smoking cessation activity. METHOD: All physicians employed by various health facilities in a community were surveyed using a structured questionnaire. Of the physicians surveyed, 523 (69.6%) returned completed questionnaires. RESULTS: The smoking rate of physicians was 29.3% (34.2% in males, 3.6% in females) and the knowledge and attitude scores to smoking were 22.5+/-2.4 and 65.4+/-6.9, respectively. The self-efficacy score was 3.4+/-1.0 and the smoking cessation activity score was 65.4+/-6.9. The smoking cessation activity was statistically significant with working place, specialty, knowledge and attitude to smoking and self-efficacy. In stepwise multiple regression, smoking cessation activity was predicted by doctors' working place, specialty, attitudes related to smoking issues, and self-efficacy of counseling knowledge and skills. CONCLUSION: Physicians need to participate routinely and actively in smoking cessation activity. For doctors to effectively counsel and intervene in patients regarding smoking cessation, it is essential to integrate education on smoking cessation intervention into curricula in formal education and to offer continuing education including smoking cessation intervention.

A clinical research of tsutsugamushi disease occurred in and around Jeoung-up in the autumn of 2000

BACKGROUND: In korea, tsutsugamushi disease is one of the common diseases which occurs in more than 40% among acute febrile diseases during Autumn. The diagnosis is confirmed with Rickettsia tsutsugamushi antibody, and is characterised by fever, chill, headache, myalgia, skin rash, escha and lymphadenopathy. METHODS: We have conducted a survey on 16 clinically and serologically confirmed cases of Tsutsugamushi disease occurring during the period of October -November, 2000. RESULTS: Of 16 cases,7 were males and 9 were females with an average age of 67.76. Most patients had fever, chill, headache, myalgia, sore throat, cojunctival injection, cough, abdominal, pain, nausea, vomiting, hematuria in order of frequency. The physical findings were eachar (81.257) and skin rash (62.77) . Serologically 7 cases (43.75%) of 16 cases were confirmed positively R.tsutsugamushi antibody. General hematologic findings were decreased platelet count (37.57) , increased or decreased WBC (31.25% and 25% each other) , and anemia (25%) , Test for liver function included elevated AST, ALT (68.76%) , alkaline phosphatase (62.6%) , hypoalbuminemia (12.6%) , and hyperbilirubinemia(6.25) . Urinalysis showed hematuria (50%) , proteinuria (50%) , and pyuria (12.67) . Doxyrcycline therapy decreased fever in 2.85 days and after 5-6 days patients were discharged with improvement of almost all symptoms. CONCLUSION: Primary care physician in a community should always consider tsutsugamushi disease when he encounters patients with acute febrile disease in late Autumn and early winter and expect good prognosis with early diagnosis and treatment.

A Study of Anger, Alexithymia, and Depression in the Functional Dyspepsia

BACKGROUND: Functional dyspepsia (FD) is a commonly encountered disturbance of gut function and has been shown to be associated with psychological disturbance such as depression and anxiety. Of particular importance to clinicians are the relationship between anger, alexithymia, and depression. In this study, we investigated anger, alexithymia, and depression in patients with functional dyspepsia. METHODS: Thirty patients who visited Wonkwang University Hospital from January 2001 to June 2001, were diagnosed with functional dyspepsia by a gastroenterologist and compared with 37 healthy control group. Medical investigation of FD including gastrofiberscopy, esophageal manometry, and ambulatory 24-hours intraesophageal reflux test were negative. All subjects were evaluated for depression, anxiety, anger and anger expression, and alexithymia. The measures included Beck Depression Inventory (BDI), Spielberger State-Trait Anxiety Inventory (STAI), Spielberger State-Trait Anger Expression Scale (STAXI), and Toronto Alexithymia Scale (TAS). RESULTS: The FD patients reported significantly more symptoms of depression, more difficulty describing feeling to others in TAS, less anger-in and anger-out expression in STAXI than the control subjects. Depressive symptoms in FD were positively correlated with state anxiety, trait anxiety, alexithymia, state anger, trait anger, and anger-in expression. In multiple regression model, state anger and trait anxiety together accounted for 69.1% of the depression in FD. CONCLUSION: The FD patients reported more depressive symptoms, and the depressive symptoms were related to anxiety, anger and anger-in, and alexithymia. These finding lend support that FD is a syndrome in which biopsychosocial process and affect dysregulation may play a role in features of FD.

A Meta-analysis on the Association between Bladder Cancer and Glutathione S transferase mu Genetic Polymorphism / 대한산업의학회지

This study was conducted to integrate the results of studies which assess the relationship between bladder cancer and Glutathione S transferase mu genetic polymorphism. We retrieved the literatures using MEDLINE search, with bladder cancer and Glutathione S transferase as key words, which were reported from 1980 to October 1998. The criteria for quality evaluation were as follows; 1) The paper should have histologically confirmed bladder cancer as case definition. 2) The paper should use the GSTM1 gene typing as method for analysis. Among 59 retrieved articles, fourteen studies were selected for quantitative meta-analysis. The overall effect size of the risk of bladder cancer due to GSTM1 was calculated by common odds ratio. Before the integration of each effect sizes into common effect sizes, the homogeneity test were conducted. All studies were case control design and cases were transitional cell carcinoma or squamous cell carcinoma of bladder. And only four papers used matching technique. Homogeneity of studies were rejected by Breslow-Day test(P<0.01), so random effect model was used for evaluation of odds ratio. The overall odds ratio of GSTM1 associated with bladder cancer was 1.55 (95% confidence interval 1.27 to 1.90) and cumulative odds ratio became more stable when the study subjects were over 1,500. Our result suggested that positive association be found between GSTM1 genetic polymorphism and bladder cancer.