Study of some Factors Influencing the Growth of Candida Albicans in Vitro

The role of carbohydrates, bacteria and bacterial supernatant in the multiplication of Candida albicans in vitro has been studied in this work. Our results show that the yeast growth: –1) is enhanced by addition to the medium of some carbohydrates, –2) is decreased by the presence of GRAM-negative bacteria, in particular Pseudomonas aeruginosa , –3) is also decreased by Ps. aeruginosa supernatant, although the exact mechanism of this phenomenon is not specified.

Zusammenfassung

Die vorliegende Arbeit untersucht den Einfluß von Kohlehydraten, Bakterien sowie der obenaufschwimmenden Schicht der entsprechenden Kulturen auf die Vermehrung von Candida albicans in vitro. Unsere Ergebnisse zeigen, daß das Hefewachstum: 1. ge-fördert wird durch Zugabe gewisser Kohlehydrate, 2. gehemmt wird durch die Gegen-wart GRAM-negativer Bakterien, insbesondere von Pseudomonas aeruginosa , 3. zu einem gewissen Grad audi gehemmt wird durch die Gegenwart von Kulturfiltrat der Ps. aeruginosa Zucht, obwohl der genaue Mechanismus dieser Hemmung nicht weiter beschrieben wird.     

黑素瘤细胞培养上清液对人单核细胞体外IL-12产生和CD14、CD1a表达的影响

目的 探讨M14黑素瘤细胞培养上清液(MCS)在体外对人外周血单核细胞分泌白介素12(IL-12)以及单核细胞向树突细胞分化的影响.方法 分离并获得人外周血单核细胞,添加不同量的MCS与单核细胞共培养,用酶联免疫吸附法(ELISA)检测单核细胞IL-12的分泌情况;将MCS与单核细胞连续5d共培养,用流式细胞仪分析单核细胞表面CD14、CD1a的表达情况.结果 MCS对单核细胞分泌活性的影响呈浓度依赖关系,随MCS添加量的逐渐增加(25-100μL),单核细胞分泌IL-12的量则呈逐渐降低趋势(158.40±21.37pg/mL-56.90±15.60pg/mL,P<0.05);同时MCS妨碍单核细胞由CD14表型(5.47%±1.00%-17.87%±2.40%,P<0.01)向树突细胞CD1a表型(41.00%±2.12%-20.00%±4.24%,P<0.01)转化;且MCS抑制单核细胞IL-12分泌和抑制单核细胞向树突细胞表型转化的作用均能被细胞外信号调节激酶(ERK)特异性抑制剂(PD98059)所逆转(分别为:172.63±8.88pg/mL,P<0.05;8.67%±0.47%,P<0.05;36.67%±0.71%,P<0.05).结论 MCS可能通过分泌ERK刺激因子对单核细胞分泌IL-12p40和单核细胞分化起到抑制作用.     

Latent 1,3-β-D-glucan acts as an adjuvant for allergen-specific IgE production induced by Japanese cedar pollen exposure

BackgroundThe pollen grains of several plant species contain 1,3-β-D-glucan (BG). BG activates dendritic cells (DCs) and subsequently regulates the innate immune responses. Within Japan, the most common disease associated with type-I hypersensitivity is Japanese cedar pollinosis. However, the role of BG in Japanese cedar pollen (JCP) remains unclear. This study examined the localization and immunological effects of BG in JCP.MethodsThe localization of BG in JCP grain was determined by immunohistochemical staining using a soluble dectin-1 protein probe and a BG recognition protein (BGRP). The content of BG extracted from JCP was measured by a BGRP-based ELISA-like assay. The cytokine production by bone marrow-derived DCs (BMDCs) obtained from wild-type and BG receptor (dectin-1) knock-out mice was examined in vitro. The mice were intranasally administered JCP grains and the specific serum Ig levels were then quantified.ResultsBG was detected in the exine and cell wall of the generative cell and tube cell of the JCP grain. Moreover, BG in the exine stimulated production of TNF-α and IL-6 in the BMDCs via a dectin-1-dependent mechanism. Meanwhile, JCP-specific IgE and IgG were detected in the serum of wild-type mice that had been intranasally administered with JCP grains. These mice also exhibited significantly enhanced sneezing behavior. However, dectin-1 knock-out mice exhibited significantly lower JCP-specific IgE and IgG levels compared to wild-type mice.ConclusionsLatent BG in JCP can act as an adjuvant to induce JCP-specific antibody production via dectin-1.     

Mutational analysis of cytocentrifugation supernatant fluid from pancreatic solid mass lesions

Diagnosis of fine‐needle aspirations of pancreatic solid masses is complicated by many factors that keep its false‐negative rate high. Our novel approach analyzes cell‐free cytocentrifugation supernatant, currently a discarded portion of the specimen. Supernatant and cytology slides were collected from 25 patients: 11 cases with confirmed outcome [five positive (adenocarcinoma) and six negative (inflammatory states)], plus 14 without confirmed outcomes. Slides were microdissected, DNA was extracted from microdissections and corresponding supernatants, and all were analyzed for KRAS point mutation and loss of heterozygosity. Notably, higher levels of free DNA were found in supernatants than in corresponding microdissected cells. Supernatants contained sufficient DNA for mutational profiling even when samples contained few to no cells. Mutations were present in 5/5 malignancies and no mutations were present in inflammatory states. In conclusion, these findings support using supernatant for mutational genotyping when diagnostic confirmation is required for pancreatic solid masses. Diagn. Cytopathol. 2014;42:719–725. © 2013 Wiley Periodicals, Inc.     

Glucocorticoid receptor number and intracellular water space

In order to elucidate the relationship between cell water content and number of glucocorticoid receptors, eleven normal and malignant lymphoid or myelomonocytic cell types originating from mouse, rat and man were investigated. The cellular water space was measured with 3H2O, and glucocorticoid receptor number was measured in a whole-cell binding assay with [3H]dexamethasone at 30 and 37 degrees C. The intracellular water phase concentration of glucocorticoid receptors (around 40 nmol/l cell water), and the dependence of receptor affinity on temperature were similar in normal and malignant rodent and human cells. It is concluded that comparisons of glucocorticoid receptor levels are best made on the basis of intracellular receptor concentrations.     

Biologic characteristics of an immunosuppressive factor derived from a human lung cancer cell line

A 549, a human lung cancer cell line, spontaneously produces a tumor-derived immunosuppressive factor (TDSF) which inhibited PHA-stimulated T lymphocyte proliferation via a noncytotoxic mechanism. The inhibition increased in a dose-dependent pattern. The factor also markedly suppressed production of interleukin (IL-2) by PHA-stimulated lymphocytes and IL 2-dependent proliferation of activated lymphocytes. The fact that TDSF possessed very potent inhibitive action on IL-2 is especially noteworthy if we consider the use of IL-2 as immunotherapeutic agent. The synthesis of the factor was inhibited by mitomycin C, actinomycin D and cycloheximide, indicating that the factor is a genic product of A 549 cells. The factor is chemically a protein with a molecular weight greater than 150 KD and sensitve to extremes of pH, heating to 60 °C and trypsin treatment.     

Nitrite binding to cytochrome P-450 from liver microsomes of trout (Salmo gairdneri Rich.) and effects on two microsomal enzymes

Addition of nitrite to dithionite-reduced trout liver microsomes leads to the conversion of cytochrome P-450 into a cytochrome P-420-NO complex, as it does in mammalian microsomes. A loss in cytochrome P-450 and an inhibition of aminopyrine demethylase (AP) activity were observed in vitro at nitrite-concentrations found in the liver of trout exposed in vivo to this toxin. Nitrite had no effect on dimethylaniline monooxygenase (DMA), a cytochrome P-450-independent enzyme.