Human eosinophils exert TNF-α and granzyme A-mediated tumoricidal activity toward colon carcinoma cells

Peripheral blood and tissue eosinophilia is a prominent feature in allergic diseases and helminth infections. In cancer patients, tumor-associated tissue eosinophilia is frequently observed. Tumor-associated tissue eosinophilia can be associated with a favorable prognosis, notably in colorectal carcinoma. However, underlying mechanisms of eosinophil contribution to antitumor responses are poorly understood. We have in this study investigated the direct interactions of human eosinophils with Colo-205, a colorectal carcinoma cell line, and show that eosinophils induce apoptosis and directly kill tumor cells. Using blocking Abs, we found that CD11a/CD18 complex is involved in the tumoricidal activity. Coculture of eosinophils with Colo-205 led to the release of eosinophil cationic protein and eosinophil-derived neurotoxin as well as TNF-α secretion. Moreover, eosinophils expressed granzyme A, which was released upon interaction with Colo-205, whereas cytotoxicity was partially inhibited by FUT-175, an inhibitor of trypsin-like enzymatic activity. Our data present the first demonstration, to our knowledge, that granzyme A is a cytotoxic mediator of the eosinophil protein arsenal, exerting eosinophil tumoricidal activity toward Colo-205, and provide mechanistic evidence for innate responses of eosinophil against tumor cells.     

Plasma interleukin-1β concentration is associated with stroke in sickle cell disease

The pathogenesis of sickle cell disease (HbSS), which has numerous complications including stroke, involves inflammation resulting in alteration of plasma inflammatory protein concentration. We investigated HbSS children with abnormal cerebral blood flow detected by trans-cranial Doppler ultrasound (TCD) who participated in the multi-center stroke prevention (STOP) study, to determine if plasma inflammatory protein concentration is associated with the outcome of stroke. Thirty-nine plasma samples from HbSS participants with elevated TCD who had no stroke, HbSS-NS (n = 13) or had stroke, HbSS-S (n = 13), HbSS steady-state controls (n = 7) and controls with normal hemoglobin, HbAA (n = 6), were analyzed simultaneously for 27 circulating inflammatory proteins. Logistic regression and receiver operating characteristics curve analysis of stroke on plasma inflammatory mediator concentration, adjusted for age and gender, demonstrated that interleukin-1β (IL-1β) was protective against stroke development (HbSS-NS = 19, 17–23, HbSS-S = 17, 16–19 pg/mL, median and 25th–75th percentile; odds ratio = 0.59, C.I. = 0.36–0.96) and was a good predictor of stroke (area under curve = 0.852). This result demonstrates a strong association of systemic inflammation with stroke development in HbSS via moderately increased plasma IL-1β concentration, which is furthermore associated with a decreased likelihood of stroke in HbSS.     

Inhibition of K+ efflux prevents mitochondrial dysfunction, and suppresses caspase-3-, apoptosis-inducing factor-, and endonuclease G-mediated constitutive apoptosis in human neutrophils

Neutrophils die rapidly via apoptosis and their survival is contingent upon rescue from constitutive programmed cell death by signals from the microenvironment. In these experiments, we investigated whether prevention of K⁺ efflux could affect the apoptotic machinery in human neutrophils. Disruption of the natural K⁺ electrochemical gradient suppressed neutrophil apoptosis (assessed by annexin V binding, nuclear DNA content and nucleosomal DNA fragmentation) and prolonged cell survival within 24–48 h of culture. High extracellular K⁺ (10–100 mM) did not activate extracellular signal-regulated kinase (ERK) and Akt, nor affected phosphorylation of p38 MAPK associated with constitutive apoptosis. Consistently, pharmacological blockade of ERK kinase or phosphatidylinositol 3-kinase (PI 3-kinase) did not affect the anti-apoptotic action of KCl. Inhibition of K⁺ efflux effectively reduced, though never completely inhibited, decreases in mitochondrial transmembrane potential (ΔΨ_m) that preceded development of apoptotic morphology. Changes in ΔΨ_m resulted in attenuation of cytochrome c release from mitochondria into the cytosol and decreases in caspase-3 activity. Culture of neutrophils in medium containing 80 mM KCl with the pan-caspase inhibitor Z-VAD-FMK resulted in slightly greater suppression of apoptosis than KCl alone. High extracellular KCl also attenuated translocation of apoptosis-inducing factor (AIF) and endonuclease G (EndoG) from mitochondria to nuclei. The DNase inhibitor, aurintricarboxylic acid (ATA) partially inhibited nucleosomal DNA fragmentation, and the effects of ATA and 80 mM KCl were not additive. These results show that prevention of K⁺ efflux promotes neutrophil survival by suppressing apoptosis through preventing mitochondrial dysfunction and release of the pro-apoptotic proteins cytochrome c, AIF and EndoG independent of ERK, PI 3-kinase and p38 MAPK. Thus, K⁺ released locally from damaged cells may function as a survival signal for neutrophils.     

The modalities of the sexual cycle of Octopus vulgaris in the southern Moroccan Atlantic (Tantan, Boujdour)

The reproduction of Octopus vulgaris between Tan Tan and Boujdour is studied. The samples used are sorted monthly from commercial catches of coastal trawlers operating in this area from December 2001 to May 2003. The study shows that sexual maturity is in advance for males than for females. Two spawning periods are made out by the follow-up of the RGS: a spring period from March to July, and an autumnal period, which is less intense, between September and October. The division of spermatogonia and spermatocytes reaches its maximal intensity in July and December, whereas the spermiogenesis is active until the time of mating. The vitellogenesis starts in mid-December and extends until the following spawning period. This reveals a later gonadic development for females than for males; approximately seven months for females and three months for males.     

Distribution et écologie des associations d’ostracodes récents de l’estuaire de Tahadart (Maroc Nord-Occidental)

Forty-five ostracod species were identified for the first time in the Tahadart estuary (NW Morocco). They can be grouped into five different ecological assemblages, which characterise four estuarine ecozones. A first ecozone occurs in temporary waters located between the tidal channels of the inner estuary and it is characterised by ostracod species typical of fresh to brackish waters. In the main estuary area, the ostracod fauna is dominated by clearly euryhaline species that account for nearly 70% of the entire fauna. The relative abundance of the different assemblages allows us to subdivide the estuarine environment along a longitudinal profile into mouth, outer and inner estuary. The estuary's mouth displays the highest values of marine euhaline, phytal and periphytal as well as ubiquitous species and the lowest values of very euryhaline species; the latter are the only species present in the inner estuary. The outer estuary exhibits intermediate percentages of the different ecological assemblages. This distribution, characterized by the absence of limnic species, points to the influence of marine waters in an open estuary with a low fresh water input. In the estuary's mouth, as well as in the outer estuary, where the slikke and schorre are well developed, the distance to the emersion line is underlined by the lower occurrence of marine and ubiquitous species and the higher occurrence of the very euryhaline ones. The Tahadart estuary displays a moderate to high ostracod diversity with distinct differences among the ecological zones. The stress of this environment is highlighted by the low diversity characteristics of extreme environments, alike in the inner estuary and in some areas located far from the emersion level. The relative position with respect to the emersion line, as well as the presence of gross detritic sediments and vegetation, appear to be of importance for the good development of the ostracods.     

Evaluation des dégâts par les vers blancs (Coleoptera : Scarabaeoidea) dans les parcelles de régéneration du chêne-liège (Quercus suber L.) en forêt de la Mamora (Maroc) et recherche de médiateurs chimiques pour une lutte biologique

Résumé

Au cours des dernières décennies, les essais de régénération du chêne-liège en forêt de la Mamora se sont heurtés à des attaques massives de vers blancs sur les racines des jeunes plants, avec un taux de réussite des plantations ne dépassant guère 12% dans la majorité des parcelles de régénération. La biologie de Sphodroxia maroccana Ley (Coleoptera : Melolonthidae), le ravageur principal, a été en partie élucidée, avec encore des lacunes concernant la période exacte ?émergence des mâles par rapport aux femelles, la longévité des imagos et la sex-ratio. La sècheresse esti vale est parmi les autres causes de dépérissement des jeunes plants. Lors de la première année suivant la plantation dans des parcelles expérimentales, la mortalité cumulée due aux attaques larvaires et à la sècheresse a varié entre 41% et 68% selon les blocs considérés dans les parcelles. La mortalité liée aux attaques des larves de S. maroccana était comprise entre 24 et 43%, avec une distribution en taches des dégâts, plus ou moins importantes selon la densité initiale des plants. L’isolement par micro-extraction en phase solide des effluves femelles de S. maroccana a permis ?identifier le résorcinol (1,3-benzènediol) comme composé phéromonal présumé. La fonction de cette molécule comme phéromone reste toutefois à démontrer.     

Detection of BrdU-label retaining cells in the lacrimal gland: implications for tissue repair

The purpose of the present study was to determine if the lacrimal gland contains 5-bromo-2′-deoxyuridine (BrdU)-label retaining cells and if they are involved in tissue repair. Animals were pulsed daily with BrdU injections for 7 consecutive days. After a chase period of 2, 4, or 12 weeks, the animals were sacrificed and the lacrimal glands were removed and processed for BrdU immunostaining. In another series of experiments, the lacrimal glands of 12-week chased animals were either left untreated or were injected with interleukin 1 (IL-1) to induce injury. Two and half days post-injection, the lacrimal glands were removed and processed for BrdU immunostaining. After 2 and 4 weeks of chase period, a substantial number of lacrimal gland cells were BrdU⁺ (11.98 ± 1.84 and 7.95 ± 1.83 BrdU⁺ cells/mm², respectively). After 12 weeks of chase, there was a 97% decline in the number of BrdU⁺ cells (0.38 ± 0.06 BrdU⁺ cells/mm²), suggesting that these BrdU-label retaining cells may represent slow-cycling adult stem/progenitor cells. In support of this hypothesis, the number of BrdU labeled cells increased over 7-fold during repair of the lacrimal gland (control: 0.41 ± 0.09 BrdU⁺ cells/mm²; injured: 2.91 ± 0.62 BrdU⁺ cells/mm²). Furthermore, during repair, among BrdU⁺ cells 58.2 ± 3.6 % were acinar cells, 26.4 ± 4.1% were myoepithelial cells, 0.4 ± 0.4% were ductal cells and 15.0 ± 3.0% were stromal cells. We conclude that the murine lacrimal gland contains BrdU-label retaining cells that are mobilized following injury to generate acinar, myoepithelial and ductal cells.