Circulating innate immune markers and outcomes in treatment-naïve advanced non-small cell lung cancer patients.

INTRODUCTION: Innate immunity represents the first step of activation of the immune system and dictates the quality of adaptive immune responses. Studies have reported links between systemic inflammatory or innate immune markers and prognosis in patients with lung cancer. To our knowledge, the prospective and concomitant study of these systemic markers has never been performed. METHODS: Advanced treatment-naive non-small cell lung cancer (NSCLC) patients eligible for first-line platinum-based chemotherapy were prospectively included from December 2012 to July 2015 (N = 148). Blood samples of patients were collected before the first cycle for fresh NK cell phenotyping. Peripheral blood mononuclear cells were cryopreserved for natural cytotoxicity receptor (NCR) genotyping as well as sera for NCR's ligand quantification. Data on leukocytes, neutrophils and monocyte counts and lactate dehydrogenase (LDH) levels were extracted from electronic medical records. RESULTS: Among all studied markers, monocytosis, neutrophilia, leucocytosis, high LDH and sBAG6 levels and reduced levels of NCR3 transcripts were associated with poor overall survival (OS) in univariate analysis. The levels of NCR3 transcripts was linked to age, number of metastatic sites, monocyte counts, LDH and sBAG6 levels. Neutrophilia was associated to high sBAG6 levels. NCR3 was the unique innate immune parameter that remained as an independent factor associated with both OS (P = 0.003) and progression-free survival (P = 0.009) in the multivariate analysis. CONCLUSION: This study brought evidence that these biomarkers are entangled; parameters associated with an inflammatory process were related to reduced levels of NCR3 transcripts. Finally, the level of NCR3 transcripts was independently associated with outcomes in treatment-naive patients with advanced NSCLC.

Bioavailability of soluble oxalate from tea and the effect of consuming milk with the tea.

OBJECTIVES: To measure the availability of oxalate normally extracted when making tea from two commercially available black teas bought from a supermarket in Christchurch, New Zealand in July 2001. DESIGN, SUBJECTS AND INTERVENTION: A randomized double crossover study. Six students and four staff consumed six cups of each brand of tea both with and without added milk over a 24 h period. A total urine collection was taken for the initial 6 h followed by a further 18 h. The oxalate content of the urine voided was measured using an enzyme kit method and the availability of the soluble oxalate consumed was measured for the 6 h and the total 24 h sample. SETTING: University campus. RESULTS: The mean soluble oxalate content of black tea in the two different commercial tea bags was respectively 6.1 and 6.3 mg soluble oxalate/g tea. The mean availability of the oxalate extracted from tea measured over a 6 h period ranged from 1.9 to 4.7% when tea was consumed without milk. The availability of the soluble oxalate from tea ranged from -3.0 to 2.3% for each of the two brands of tea investigated over a 24 h period. CONCLUSION: These studies show that consuming black tea on a daily basis will lead to a moderate intake of soluble oxalate each day, however the consumption of tea with milk on a regular basis will result in the absorption of very little oxalate from tea.

Poaceae in the natural diet of the snail Helix aspersa Müller (Gastropoda, Pulmonata).

The natural diet of Helix aspersa was studied in two populations by analysing faeces. Picris echioides, Carduus tenuifloris, Urtica dioica, Galium molugo (Dicotyledons) and Poaceae (Monocotyledons) were the principal resources of the studied populations. The species of Poaceae ingested by Helix aspersa in July were determined by analysing the phytoliths present in the faeces. Festuca rubra, Dactylis glomerata and Bromus hordeaceus seemed to be attractive for the snails whereas Elytrigia repens was rejected. These results were convergent with the ingestion rates of Poaceae in the laboratory but assimilation efficiencies did not explain these choices. Poaceae with the highest energetic values were not preferred to others. The texture, the mineral and organic contents of the Poaceae may be responsible for the snails feeding choices. The importance of Poaceae for this species is discussed.

Mannan-degrading enzymes purified from the crop of the brown garden snail Helix aspersa Müller (Gastropoda Pulmonata).

Two mannan-degrading enzymes were purified from the crop of the terrestrial snail Helix aspersa Müller. The crude extracts were taken from dormant (for 4 months) snails. The enzymes were a betaD-mannanase of 37.4 +/- 0.3 kDa (EC 3.2.1.78) and a betaD-mannosidase of 77.8 +/- 1.9 kDa (EC 3.2.1.25). Both enzymes degraded insoluble mannan, releasing either mannose only (beta-mannosidase) or oligosaccharides, possibly mannotriose and mannopentaose (beta-mannanase). The beta-mannanase had a typical endo-activity pattern, while the beta-mannosidase was an exoenzyme. The incubation of both enzymes with mannan increased the catalysis by 83%. The best synergy was found with 75% mannosidase combined with 25% mannanase. The beta-mannanase also hydrolysed beta-linked heteromannans and its affinity for different galactomannans was studied. The Km values, varying from 2.89 +/- 0.47 mg x ml(-1) to 0.26 +/- 0.01 mg x ml(-1), revealed the inhibitory effect of the alphaD-galactosyl residues released. The beta-mannosidase was acidic (optimum pH = 3.3) and heat-sensitive (50% residual activity at 42 degrees C after 5 min of pre-incubation), while the beta-mannanase remained stable until 48.5 degrees C (50% residual activity) and over a pH range of 4.3-7.5. The properties of these mannanolytic enzymes are discussed in this paper compared with those purified in other gastropods and in a bacterium, Enterococcus casseliflavus, a quite similar strain previously isolated from this snail intestine. The occurrence of thermostable enzymes in H. aspersa digestive tract could be a zootechnic parameter of great importance for snail farming. J. Exp. Zool. 290:125-135, 2001.

First characterization of 6-hydroxytryptamine in the rat midbrain by using specific antibodies.

The visualization of serotonin, 5-methoxytryptamine, and tryptamine in the rat midbrain has been made possible by the development of antibodies raised against these conjugated molecules. It has been suggested that 6-hydroxytryptamine (6-HT) might also be a neurotransmitter in this region. To test this hypothesis, 6-HT was synthesized and antibodies were raised in the rabbit. The high avidity (IC50 = 5 x 10(-9) M) and specificity [cross-reactivity ratio between 6-HT-glutaraldehyde (G)-bovine serum albumin (BSA) and 5-HT-G-BSA, the most immunoreactive compound, was 1,500] rendered these antibodies reliable tools for specific molecular detection of 6-HT in the G-fixed tissues. In the dopaminergic region, 6-HT immunoreactivity was noted in the substantia nigra but was particularly intense in the red nuclei, where it seems to be localized in the magnocellular division in the form of large 6-HT neurons. In contrast, there were few 6-HT neurons in the raphe nuclei. Thus, 6-HT may be a new putative neurotransmitter existing in the red nuclei, in addition to the other neurotransmitters already described in this region, in the nigro-rubral pathway, and in the rubral projection from the dorsal raphe nuclei. 6-HT is possibly implicated in motor control and might exert hallucinogenic properties as do other 6-hydroxylated indoleamines.

Autoantibodies directed against lipid membrane components in sera of patients with malignant tumors.

Tumor-associated antigens (Ag) are expressed on neoplastic cells. Using an enzyme-linked immunosorbent assay (ELISA), an attempt was made to evaluate the autoimmune responses directed against lipid membrane components. A comparison was made of autoantibody (autoAb) levels in human sera of 684 patients with malignant tumors and those of 185 controls (healthy subjects and patients suffering from other diseases). A highly significant difference was found between the immunological binding of the groups for only one phospholipids (PL), i.e., phosphatidylinositol (PI). Using ELISA tests and PI-related compounds differing in fatty acid residue types and/or in phosphatidyl group, it was demonstrated that the hydrophilic residue is the immunodominant part recognized by the autoAb detected in sera of cancer patients.

Monoclonal anti-conjugated acetylcholine antibody and immunohistochemical applications in rat nervous system.

Acetylcholine (ACh) conjugates were injected into AKR and DBA mice over a period of 10 weeks. The polyclonal antisera were tested at various immunization times for affinity and specificity using an enzyme-linked immunosorbent assay (ELISA). The most immunoreactive compound was found to be choline-glutaryl-bovine serum albumin (or conjugated ACh). The AKR and DBA mice yielding the highest apparent affinity were killed, and the spleen cells were fused with X63 or SP2/O/Ag mouse myeloma cells. Supernatants of confluent cultures were tested for the presence of anti-conjugated ACh antibodies using the same ELISA method. The best results were obtained with the hybridomas from AKR spleen cells and X63 mouse myeloma cells. Monoclonal antibody affinity and specificity were then evaluated by a radioimmunological procedure using iodinated monoclonal anti-conjugated ACh antibody. From competition experiments, the most immunoreactive compound was choline-glutaryl-protein. The other related compounds were recognized either poorly or not at all. The high affinity and specificity of our monoclonal antibody enabled us to visualize ACh molecules on fixed rat brain sections. ACh was fixed with a mixture of nitrobenzyl alcohol and glutaraldehyde. Many ACh-immunoreactive cell bodies and fibers were seen on sections from the basal forebrain and spinal cord. Preadsorption and other immunohistochemical tests demonstrated that the ACh staining was highly specific.

[Food consumption, production and energy evaluation in Helix aspersa müller (a terrestrial pulmonated gasteropod)]. / Consommation alimentaire, production et bilan énergétique chez Helix aspersa müller (gastéropode pulmoné terrestre).

A quantitative study of feeding in Helix aspersa Müller (Gastropod, Pulmonate) is made in the laboratory with a view to apply the resulted obtained to its rearing in a warmed room. From the birth of the snails to six months old, two methods are applied: one ponderous, the other energetic, allowing us to draw up a nutritional budget. --The parameters are: daily rates of consommation, fecal production and assimilation; monthly production rate, efficiency of assimilation and finally gross and net growth efficiencies. The evolution of these parameters, seems to be dependant on the stage of growth of the snail, and on a physiological phenomenon present in Helix aspersa living in the field.

[Nitrogen excretion of Helix aspersa Müller (Mollusca Gastropoda Pulmonata)]. / Contribution a l'étude de l'excrétion azotée chez Helix aspersa Müller (Mollusque gastéropode pulmoné).

Helix aspersa, studied in Brittany, has not a complete uricolytic enzymatic chain. Allantoïnase and allantoïcase are absent, but, in vitro, uricase and urease are very active. A part from uric acid, Helix aspersa shows a high rate of urea and ammonia. The level of these substances is dependant on the age of the snail and on its physiological state.