An optimized cocktail of small molecule inhibitors promotes the maturation of dendritic cells in GM-CSF mouse bone marrow culture.

Dendritic cells (DCs) are the most potent antigen-presenting cells, playing an essential role in the pathogen and tumor recognition, and anti-tumor immunity, and linking both the innate and adaptive immunity. The monocyte-derived DCs generated by ex vivo culture, have been used for cancer immunotherapy to eliminate tumor; however, the clinical efficacies are not sufficient, and further improvement is essential. In this study, we established a method to generate DCs using small molecule compounds for cancer immunotherapy. We observed an increase in the percentage of CD11c+I-A/I-Ehigh cells, representing DCs, by adding four small molecular inhibitors: Y27632, PD0325901, PD173074, and PD98059 (abbreviated as YPPP), in mouse bone marrow (BM) culture with granulocyte-macrophage colony stimulating factor (GM-CSF). BM-derived DCs cultured with YPPP (YPPP-DCs) showed high responsiveness to lipopolysaccharide stimulation, resulting in increased interleukin (IL) -12 production and enhanced proliferation activity when co-cultured with naïve T cells compared with the vehicle control. RNA-seq analysis revealed an upregulation of peroxisome proliferator - activated receptor (PPAR) γ associated genes increased in YPPP-DCs. In tumor models treated with anti-programmed death (PD) -1 therapies, mice injected intratumorally with YPPP-DCs as a DCs vaccine exhibited reduced tumor growth and increased survival. These findings suggested that our method would be useful for the induction of DCs that efficiently activate effector T cells for cancer immunotherapy.

PCBP1 acts as a regulator of CCL2 expression in macrophages to induce recruitment of monocyte-derived macrophages into the inflamed colon.

Intestinal macrophages with functional plasticity play essential roles in gut immune responses by increasing chemokines and cytokines, thereby contributing to the pathogenesis of inflammatory bowel disease (IBD). Poly(rC)-binding protein 1 (PCBP1), which is widely expressed in immune cells, binds to nucleic acids in mRNA processing, stabilization, translation and transcription. However, little is known about the influence of PCBP1 on macrophages and its specific mechanism in inflamed intestines. In this study, conditional depletion of Pcbp1 in macrophages protected mice from progression of dextran sulfate sodium induced colitis and resulted in significant alleviation of colitis. Pcbp1 deficiency markedly decreased C-C motif chemokine ligand 2 (CCL2) production by colonic CX3C motif chemokine receptor 1+ (CX3CR1+) macrophages and reduced accumulation of pro-inflammatory macrophages and production of pro-inflammatory cytokines, such as IL-6 and TNF-α, in the inflamed colon. RNA-immunoprecipitation analysis indicated that PCBP1 might interact with Ccl2 mRNA and regulate its expression in macrophages. PCBP1 expression in inflamed intestines also correlated significantly with IBD severity in patients, suggesting a critical involvement of PCBP1 in intestinal inflammation. We anticipate that our findings will facilitate the development of novel therapeutic approaches for IBD by targeting the specific function of immune cells in the local microenvironment, thereby helping to reduce adverse effects.

A Novel CD135+ Subset of Mouse Monocytes with a Distinct Differentiation Pathway and Antigen-Presenting Properties.

The mononuclear phagocyte system (MPS), composed of monocytes/macrophages and dendritic cells (DCs), plays a critical role at the interface of the innate and adaptive immune systems. However, the simplicity of MPS has been challenged recently by discoveries of novel cellular components. In the current study, we identified the CD135+ subset of monocytes as a novel class of APCs in mice. CD135+ monocytes were readily found in the bone marrow, spleen, and peripheral blood at steady state, and they expressed markers specific to DCs, including MHC class II and CD209a, along with markers for monocytes/macrophages. In addition, this subset phagocytosed bacteria and activated naive T lymphocytes, fulfilling the criteria for APCs. CD135+ monocytes were derived directly from macrophage DC progenitors, not from common monocyte progenitors or other monocytes, suggesting that these are distinct from conventional monocytes. These findings facilitate our understanding of the MPS network that regulates immune responses for host defense.

Effects of Moxibustion on Stress-Induced Delayed Gastric Emptying via Somatoautonomic Reflex in Rats.

Objective: Moxibustion (MOX) is used to treat a wide variety of disorders, including those with gastric symptoms. However, the exact mechanisms underlying the beneficial effects of MOX are unknown. The purpose of this study was to investigate if application of indirect MOX (iMOX) to ST 36 reduces restraint stress (RS)-induced alteration in gastric responses of conscious rats, and if a somatoautonomic reflex mediates gastric emptying (GE). Materials and Methods: One group of rats was fed solid food after 24 hours of fasting. Immediately after food ingestion. These rats were subjected to RS. Ninety minutes after feeding, the rats were euthanized, and their gastric contents were removed to calculate GE. iMOX had been performed at ST 36 bilaterally throughout the stress loading. To investigate if vagal-nerve activity was involved in mediating the stress-induced alterations of GE by iMOX, atropine was intraperitoneally administered to other rats just before initiating RS; bilateral truncal vagotomy had been performed on day 14 before GE measurement. Results: RS delayed GE significantly (42.9 ± 5.8%)in stressed rats, compared to nonstressed rats (68.7 ± 1.8%). iMOX at ST 36 reduced stress-induced inhibition of GE significantly (67.1 ± 2.4%). MOX-mediated reduction of GE disappeared upon atropine injection and vagotomy. Conclusions: RS-induced delayed GE may be ameliorated by iMOX at ST 36. Somatoautonomic, reflex-induced vagal-nerve activity helps mediate the stimulatory effects of iMOX on RS-induced delayed GE. As a complementary and alternative medicine, iMOX may also be advantageous for patients with gastric disorders, such as functional dyspepsia.

Calcium Pyrophosphate Dihydrate Crystals Increase the Granulocyte/Monocyte Progenitor (GMP) and Enhance Granulocyte and Monocyte Differentiation In Vivo.

Calcium pyrophosphate dihydrate (CPPD) crystals are formed locally within the joints, leading to pseudogout. Although the mobilization of local granulocytes can be observed in joints where pseudogout has manifested, the mechanism of this activity remains poorly understood. In this study, CPPD crystals were administered to mice, and the dynamics of splenic and peripheral blood myeloid cells were analyzed. As a result, levels of both granulocytes and monocytes were found to increase following CPPD crystal administration in a concentration-dependent manner, with a concomitant decrease in lymphocytes in the peripheral blood. In contrast, the levels of other cells, such as dendritic cell subsets, T-cells, and B-cells, remained unchanged in the spleen, following CPPD crystal administration. Furthermore, an increase in granulocytes/monocyte progenitors (GMPs) and a decrease in megakaryocyte/erythrocyte progenitors (MEPs) were also observed in the bone marrow. In addition, CPPD administration induced production of IL-1ß, which acts on hematopoietic stem cells and hematopoietic progenitors and promotes myeloid cell differentiation and expansion. These results suggest that CPPD crystals act as a "danger signal" to induce IL-1ß production, resulting in changes in course of hematopoietic progenitor cell differentiation and in increased granulocyte/monocyte levels, and contributing to the development of gout.

Measuring diagnostic competency and the analysis of factors influencing competency using written case studies.

PURPOSES: To measure nurses' diagnostic competencies by using written case studies and to analyze factors influencing these competencies. METHODS: A descriptive study was conducted using survey methods and a mailed questionnaire with a convenience sample of 376 nurses. Two written case studies were used to measure the diagnostic competencies of the subjects. Possible factors influencing nurses' judgments in making nursing diagnoses that were measured were clinical experience, decision-making responsibility, knowledge of nursing diagnosis, and attitude toward the use of nursing diagnosis. FINDINGS: Subjects who demonstrated all three competencies were 35% (n = 131) for case study 1, and 53% (n = 200) for case study 2. Subjects who demonstrated diagnostic competency in defining diagnostic labels had the longest length of clinical experience. Subjects who were primary nurses with the usual responsibility of identifying nursing diagnoses showed the highest percentage of competency rates for defining characteristics, related factors, and risk factors. CONCLUSIONS: Many of these Japanese nurses did not perform at satisfactory levels of diagnostic competency with these two written case studies. Factors that influenced diagnostic competency were length of clinical experience, decision-making responsibility, and frequency of studying nursing diagnosis.

Determination method for steroid 5alpha-reductase activity using liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry.

A determination method for steroid 5alpha-reductase activity using liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry (LC/APCI-MS) in the positive-ion mode has been developed. The rat prostatic enzyme source was used and the enzymatically formed 5alpha-dihydrotestosterone and 5a-androstane-3alpha,17beta-diol were determined by LC/APCI-MS using absolute calibration curve method. The sum of the formed products was used as a measurement of the enzyme activity. This method was applied to kinetic study of this enzyme and inhibitory experiments using Finasteride as a model inhibitor.

Nursing diagnoses and interventions of Japanese patients with end-stage breast cancer admitted for different care purposes.

PURPOSE: To clarify actual nursing diagnoses for and interventions given to patients with end-stage breast cancer admitted for different care purposes. METHODS: Nursing diagnoses, defining characteristics, related/risk factors, and nursing interventions were analyzed in a convenience sample of 150 patient records. FINDINGS: A total of 539 nursing diagnoses (96 labels) were documented. Frequently listed diagnoses were chronic pain, risk for infection, and activity intolerance. The most frequently used nursing diagnosis for the chemotherapy group was risk for infection. The nurses in this study rarely report any diagnoses related to death and dying. CONCLUSIONS: Nursing diagnoses and interventions differed depending on the purpose of admission. PRACTICE IMPLICATIONS: Oncology nurses need to consider the reasons for admission when making nursing diagnoses and interventions for patients with end-stage breast cancer.

Online exclusive: family satisfaction with perception of and barriers to terminal care in Japan.

PURPOSE/OBJECTIVES: To examine how families of patients with cancer perceive and are satisfied with terminal care given in a university hospital and to identify barriers to the best care for patients and proper support for their families. DESIGN: Quantitative, descriptive. SETTING: A Japanese national university hospital. SAMPLE: Convenience sample of 73 family members of deceased patients with cancer (response rate was 55%). METHODS: Completion of a questionnaire comprised of 23 items, including multiple-choice and open-ended questions. MAIN RESEARCH VARIABLES: Satisfaction with terminal care, misunderstanding of narcotics use, recognition of disease signs and symptoms, and expectations for terminal care. FINDINGS: Ninety percent of respondents were satisfied with nursing care. Eighty percent were satisfied with the information they received at admission, 70% were satisfied at the critical phase, and 63% were satisfied at the end of life. The main expected types of desirable terminal care were pain management and spiritual care. The bereaved had difficulty dealing with patients' pain, dyspnea, and appetite loss. Forty-four patients were treated with narcotics. Fifty-seven percent of the family members of these patients found pain to be the most difficult symptom to deal with. Fifty-eight percent of the respondents believed that if narcotics were used, patients would soon die. CONCLUSIONS: Terminal cancer care given in Japanese university hospitals does not appear to be adequate in terms of patients' and families' quality of life. The findings indicate that insufficient current information is given throughout the terminal stage of cancer and patients and their families misunderstand the use of narcotics. IMPLICATIONS FOR NURSING: Oncology nurses need to consider different cultural backgrounds when offering specific care to patients with terminal cancer. Patients and their families must be educated about the efficacy of narcotics in the care of terminally ill patients with cancer.