Chemical and isotopic characterization of the thermal fluids emerging along the North-Northeastern Greece.

Hydrochemical and isotopic characteristics of fluids from major geothermal fields of middle/low temperature in N/NE Greece are examined [basins: Strymon River (SR), Nestos River Delta (ND), Xanthi-Komotini (XK), Loutros-Feres-Soufli (LFS) and Rhodope Massif]. The geodynamic context is reflected to isotopic/chemical composition of fluids, heat flow values and elevated CO2 concentrations in emitted fluids. B and Li are derived from leaching of the geothermal systems hosting rocks. δ18OH2O, δ18OSO4, δ13CCO2 values and chemical compositions of Cl, B and Li of geothermal discharges suggest two distinct source fluids. Fluids in SR exhibit high B/Cl and Li/Cl ratios, suggesting these constituents are derived from associated magmas of intermediate composition (andesitic rocks). Geothermal discharges in LFS exhibit low B/Cl and Li/Cl ratios, implying acid (rhyolitic) magmatism. δ13CCO2 and CO2/(CO2 + 105He) ratios in the west part, suggest fluids affected by addition of volatiles released from subducted marine sediments. For the eastern systems, these ratios suggest gas encountered in systems issued from mixing of crustal and mantle-derived volatiles. Isotopic geothermometers reflect, for the same direction, equilibrium processes more (LFS, XK) or less (SR) pronounced and discriminate the geothermal field from low to middle [SR, ND (Erasmio)] and middle to high enthalpy [ND (Eratino), LFS, XK].

Isotopic modeling of the sub-cloud evaporation effect in precipitation.

In dry and warm environments sub-cloud evaporation influences the falling raindrops modifying their final stable isotopic content. During their descent from the cloud base towards the ground surface, through the unsaturated atmosphere, hydrometeors are subjected to evaporation whereas the kinetic fractionation results to less depleted or enriched isotopic signatures compared to the initial isotopic composition of the raindrops at cloud base. Nowadays the development of Generalized Climate Models (GCMs) that include isotopic content calculation modules are of great interest for the isotopic tracing of the global hydrological cycle. Therefore the accurate description of the underlying processes affecting stable isotopic content can improve the performance of iso-GCMs. The aim of this study is to model the sub-cloud evaporation effect using a) mixing and b) numerical isotope evaporation models. The isotope-mixing evaporation model simulates the isotopic enrichment (difference between the ground and the cloud base isotopic composition of raindrops) in terms of raindrop size, ambient temperature and relative humidity (RH) at ground level. The isotopic enrichment (Δδ) varies linearly with the evaporated raindrops mass fraction of the raindrop resulting to higher values at drier atmospheres and for smaller raindrops. The relationship between Δδ and RH is described by a 'heat capacity' model providing high correlation coefficients for both isotopes (R(2)>80%) indicating that RH is an ideal indicator of the sub-cloud evaporation effect. Vertical distribution of stable isotopes in falling raindrops is also investigated using a numerical isotope-evaporation model. Temperature and humidity dependence of the vertical isotopic variation is clearly described by the numerical isotopic model showing an increase in the isotopic values with increasing temperature and decreasing RH. At an almost saturated atmosphere (RH=95%) sub-cloud evaporation is negligible and the isotopic composition hardly changes even at high temperatures while at drier and warm conditions the enrichment of (18)Ο reaches up to 20‰, depending on the raindrop size and the initial meteorological conditions.

O-H-C isotope ratio determination in wine in order to be used as a fingerprint of its regional origin.

Stable isotopes have been applied to determine the origin assignment and verify the geographical provenance that is considered important characteristics of wine products both for consumers and the international regulations, of wines. Stable isotope analyses of (18)O/(16)O, D/H and (13)C/(12)C ratio for the detection of origin and of adulteration in wine are discussed in this study. The δ(13)C analysis of ethanol and wines water δ(18)O underlines the importance of the photosynthetic pathway and the environmental conditions of wine. Also we discuss the main factors that are responsible for the differentiation of the oxygen isotope ratios of wine water. Data interpretation demonstrated the efficacy of δ(18)O analysis not only in the wine but also in grape berries, preferably if the determination of the δ(18)O value is employed together with the determination of the δ(2)H isotope content of wine, for the detection of the geographical origin of wine.

Toll-like receptor 4 promotes control of Leishmania infantum infection through inducement of leishmanicidal activity in host macrophages: role of mitogen activated kinases.

Establishment of Leishmania infection inside macrophages requires deactivation of various signaling pathways that are dispensable for effective immune responses against the parasite. In the present study, we provide evidence that Leishmania infantum promastigotes attachment on the surface of peritoneal macrophages, internalization and transformation to amastigotes abrogated the activation of extracellular signal-regulated protein kinases (ERK) 1/2, p38 mitogen activated protein kinases (MAPK) and c-Jun NH2-terminal kinases (JNK) and the production of pro-inflammatory cytokines IL-12 and TNFalpha. Subsequent macrophage stimulation with lipopolysaccharide (LPS) during the first hours of exposure to parasite or infection resulted in restoration of MAPK phosphorylation. However, LPS-mediated MAPK activation required parasite internalization (uptake) since cytochalasin-D pretreated macrophages did not responded to LPS stimulation. IL-12, TNFalpha, and NO production was positively regulated with MAPK phosphorylation in contrast to nuclear factor-kappaB (NF-kB) which was MAPK independent. Specifically, inhibition of MAPK activation with specific inhibitors revealed that IL-12 production required p38 MAPK activation, whereas TNFalpha and NO production required all three MAPK. The restoration of NO production resulted in decrease of infection rates. Hence, these results suggest that in contrast to phagocytosis of L. infantum promastigotes, establishment of infection does not desensitize macrophages to subsequent stimulation with LPS, resulting in parasite elimination through MAPK and NF-κB activation and partial restoration of IL-12, TNFalpha and NO synthesis.

Isotopic analysis for degradation diagnosis of calcite matrix in mortar.

Mortar that was used in building as well as in conservation and restoration works of wall paintings have been analysed isotopically (delta(13)C and delta(18)O) in order to evaluate the setting environments and secondary processes, to distinguish the structural components used and to determine the exact causes that incurred the degradation phenomena. The material undergoes weathering and decay on a large proportion of its surface and in depth, due to the infiltration of water through the structural blocks. Mineralogical analysis indicated signs of sulphation and dissolution/recrystallisation processes taking place on the material, whereas stable isotopes provided information relative to the origin of the CO(2) and water during calcite formation and degradation processes. Isotopic change of the initial delta(13)C and delta(18)O in carbonate matrix was caused by alteration of the primary source of CO(2) and H(2)O in mortar over time, particularly by recrystallisation of calcite with porewater, evaporated or re-condensed water, and CO(2) from various sources of atmospheric and biogenic origin. Human influence (surface treatment) and biological growth (e.g. fungus) are major exogenic processes which may alter delta(18)O and delta(13)C in lime mortar.

Study on distribution and origin of boron in groundwater in the area of Chalkidiki, Northern Greece by employing chemical and isotopic tracers.

This paper presents an integrate study on the occurrence and distribution of boron in groundwater in the area of Chalkidiki, Northern Greece. Groundwater samples were collected from wells used for drinking and irrigation purposes. Samples were analysed for boron, various physicochemical parameters (T, pH, EC, Ca, Mg, Na, K, Br, Cl, HCO(3), SO(4), NO(3) and As) and isotopes ((18)O, (2)H, (11)B). Boron showed high spatial variation ranged from 0.04 to 6.5mg/L. Almost 60% of the examined wells exhibited boron concentration higher than the limit of 1mg/L proposed for water intended for human consumption. The higher concentrations were determined in geothermal waters with relatively high temperature. Correlation analysis and hierarchical cluster analysis were employed to find out possible relationships among the examined parameters and groundwater samples. Chemical and isotopic fingerprints have been used to investigate the origin of boron.

Antiparasitic and immunomodulatory effect of innovative treatments against Myxobolus sp. infection in Diplodus puntazzo.

The potential antiparasitic and immunomodulatory effect of three treatments against myxosporean parasites on the innate immune system of sharpsnout sea bream (Diplodus puntazzo) was investigated. Fish naturally infected with Myxobolus sp. (Bivalvulida/Platysporina), a histozoic parasite mainly affecting the renal interstitial tissue, were treated by oral administration of a combination of salinomycin with amprolium, Origanum essential oil or fumagillin in a small-scale field trial. Various leucocyte functions influenced by myxosporean infection were examined in order to determine treatment effects on leucocyte immunocompetence of treated fish. One month post treatment all drugs caused a significant decrease in prevalence and intensity of infection in comparison to untreated, infected fish. The effect was most prominent in salinomycin with amprolium treated fish, which 1-month post treatment contained either no cysts at all or a few spores free in melanomacrophage centres revealing almost total elimination of the parasite and the antiparasitic action of the treatment. There was no histopathological evidence of drug toxicity. Antiparasitic action was accompanied by a significant enhancement of phagocytic activity demonstrated by ingestion of large numbers of latex beads and the secretion of high levels of reactive nitrogen intermediates by phagocytes in vitro. Complete restoration of the diminished mitogenic responses and serum lysozyme secretion was also detected in salinomycin with amprolium-treated fish compared to untreated, infected fish. These data suggest that salilomycin with amprolium may be a promising treatment for myxosporean infections in intensively cultured warm-water fish, exhibiting action partially via the enhancement of host, innate immune functions and leading to parasite elimination.

The impact of a successful anti-myxosporean treatment on the phagocyte functions of juvenile and adult Sparus aurata L.

The aim of the present study was to investigate the impact of a successful anti-myxosporean medication on the innate immune system of fish intensively cultured in the Mediterranean basin. For this purpose, juvenile and adult gilthead seabream (S. aurata L.) naturally infected with Polysporoplasma sparis in the kidney were used in a small-scale field trial. The infected fish were treated orally with the combination of salinomycin and amprolium, two drugs well known for their anti-coccidial effect in other animals. Drug efficacy and safety was evaluated in terms of changes observed in histopathology, mortality and P. sparis intensity and prevalence rate. Phagocytic functions of head-kidney leucocytes were also investigated at the end as well as one month post the medication. Salinomycin with amprolium exhibited a significant reduction in intensity and prevalence rate in both juvenile and adult fish, and no histopathological evidence for toxic side effects was observed. In addition, the successful treatment was closely correlated with a complete restoration of the diminished phagocytic ability and capacity as well as NO, and lysozyme secretion in a time dependent manner. This data suggests that salilomycin with amprolium can be an alternative treatment for myxosporean infections in tropical fish, possibly exhibiting their action through the enhancement of host innate functions.

Dendritic cells pulsed with peptides of gp63 induce differential protection against experimental cutaneous leishmaniasis.

The need for a vaccine against Leishmania spp., a major cause of worldwide morbidity and mortality, is urgent. We tested the efficacy of an experimental vaccination in murine models of cutaneous leishmaniasis, using dendritic cells (DCs) pulsed with synthetic or native parasite antigens. DCs pulsed with peptide 154-169aa of gp63 or soluble promastigote lysate (SPL) triggered antigen-specific immune responses and efficiently reduced lesion formation and parasite load of genetically susceptible BALB/c mice infected with Leishmania major. This effect was accompanied by a modulation of the cellular immune response towards a Th1 profile. Vaccination of genetically resistant CBA mice with DCs pulsed with peptide 154-169aa or SPL did not affect the course of the disease, whereas pulsing with the epitope 467-482aa of gp63 resulted in disease exacerbation, accompanied by a switch to a Th2 profile. In view of our continuously growing knowledge about the immunobiology of DCs, these findings suggest that vaccination with DCs pulsed with defined peptides could be a strategy against infectious diseases. Peptide selection is a prerequisite as they can differentially regulate the type of immune response in susceptible or resistant hosts.

T cell help is required to induce idiotypic-anti-idiotypic autoantibody network after immunization with complementary epitope 289-308aa of La/SSB autoantigen in non-autoimmune mice.

Immunotherapies against autoimmune diseases have been of limited success. Preventive vaccines could be developed on the basis to abrogate unwanted immune responses to defined autodeterminants. In this study it is shown that immunization of BALB/c mice with two linear T and B cell epitopes of the human La/SSB autoantigen (spanning the regions 289-308aa and 349-364aa) and their complementary forms specified by the complementary mRNA, results in characteristic B and T cell responses. Mice immunized with the 289-308aa epitope or its complementary peptide elicited specific antibodies against both epitopes. In contrast, mice immunized with the 349-364aa epitope or its complementary peptide mounted antibody titres against the immunizing peptide only. According to these data, the 289-308aa epitope and its complementary form were capable to generate an idiotypic-anti-idiotypic response, which were cross-regulated. Peptide-specific T cell proliferation and cytokine production in vitro revealed the induction of a two-stage T helper response (Th1-->Th2 type) after immunization with either the epitope 289-308 or its complementary peptide. IgG1 was the predominant subclass after immunization with the two forms of epitopes 289-308 and 349-364, while a response of the IgG2b > IgG2a was obtained after the immunization with the complementary form of 349-364 epitope reflecting the TH2/TH1 polarization, respectively. Our data suggest that the complementary peptides of two immunodominant epitopes of human LaSSB can mimic the autoantibodies against these epitopes and establish an active idiotypic-anti-idiotypic network.

Efficacy and toxicity of orally administrated anti-coccidial drugs for innovative treatments of Myxobolus sp. infection in Puntazzo puntazzo.

This study tested drugs and therapeutic compounds to determine effective commercial treatment for fishes infected with myxosporeans. Two series of shore-based experiments and 1 field trial were performed. For the shore-based experiments we used Puntazzo puntazzo (ca. 20 g weight) with kidneys infected with Myxobolus sp. Initially, 6 different doses of Fumagillin, 2 doses of Toltrazuril, and 1 dose of Amprolium, ESB3 and Salinomycin were tested. In the second shore-based experiment, infected fish were treated with Origanum essential oils, Toltrazuril with propylene glycol, Amprolium, and a combination of Salinomycin 12% + Amprolium (SA). In the field trial, P. puntazzo (ca. 165 g) infected with the parasite were treated with SA, Origanum essential oils and Fumagillin. In all trials, the drugs were added to the feed and administered according to the selected regimen. Their efficacy was evaluated in terms of mortality (acceptable level was <3%), pathology and prevalence rate of Myxobolus sp. Lesions were observed only in fish treated with Fumagillin and Toltrazuril. Pathology due to treatment with Fumagillin was observed only at doses > 6 mg kg(-1) body wt for 6 wk in the interstitial renal tissue, where slight inflammation arose. The highest dose tested (25 mg kg(-1)) also produced necrosis in the interstitial tissue, degeneration of the epithelial cells of the tubules and a reduction in melanomacrophage centre numbers. The SA combination proved the most effective treatment for Myxobolus sp. infection of P. puntazzo as (1) the therapeutic regimen and commercial product was not toxic and (2) a significant reduction occurred in the prevalence rate.

Asymptomatic canine leishmaniasis in Greater Athens area, Greece.

Leishmania (L.) infantum is the etiological agent of human and canine visceral leishmaniasis in the Mediterranean subregion. Domestic dogs are the main reservoir of the parasite in most urban areas. A survey of 1638 asymptomatic dogs registered in Greater Athens area was carried out in the Hellenic Pasteur Institute during the period 1986-1994 to investigate the prevalence of canine visceral leishmaniasis in apparently healthy dogs. Dog sera was tested using the indirect fluorescent antibody technique (IFAT). Of the 1638 dogs, 366 (22.4%) had anti-Leishmania infantum antibodies at titre greater than or equal to 1/200 which were considered positive; 53 (3.2%) had antibody titres of 1/100 and were considered uncertain; and 1219 (74.4%) dogs were seronegative. From the 366 seropositive dogs, 212 were positive at 1/1600 serum dilution, 57 at 1/800, 38 at 1/400 and 59 at 1/200. The results were plotted according the site of residence, breed and age. The rate of asymptomatic infections with L. infantum dogs in Greater Athens area appears to be significantly high. Although there is an apparent lack of clinical symptoms in these dogs, asymptomatic animals harbor a chronic L. infantum infection and as such consist a 'dangerous' reservoir with regard to the spread of the disease.

IgG, IgG1 and IgM response in Trichinella spiralis-infected mice treated with 4-deoxypirydoxine or fed a Vitamin B6-deficient diet.

The aim of this study was to investigate the effect of pyridoxine (Vitamin B6) deficiency on the immunological response of BALB/c mice infected with the parasite T. spiralis. Specific anti-parasite IgM and IgG immunoglobulins were detected by ELISA method in the serum of treated animals at different periods for 60 days post infection. Vitamin B6-deficiency was induced in two separate groups of mice by either (1) maintaining the mice on a Vitamin B6-deficient synthetic pellet diet for 40 days before infection, or (2) by daily intraperitoneal injection of 8 x 10(5) M/100 microl of 4-Deoxypyridoxine (4-DPD), a potent antagonist of Vitamin B6 for 20 days prior to infection. These two groups of mice were then injected with 100 larvae (L1-T. spiralis) per os. Parasite burdens in the mice were observed by light microscopy. Cysts were present in the diaphragms of the mice after 60 days post-infection. Parasite specific IgG, as well as IgG1 levels were determined in the sera of infected mice fed a normal diet. These levels were found to be lower in the 4-DPD-treated mice compared to the untreated mice. The inhibition started from the 10th day and continued to the 60th day, and in the 4-DPD-treated group the inhibition initiated after 24 h to 60 days. IgM level also was depressed by 4-DPD, starting from 24 h after injection of the compound. In mice fed Vitamin B6-deficient diets the levels of IgG were lower than in mice fed normal diets. These results show that BALB/c mice infected with T. spiralis and fed either a Vitamin B6-deficient diet or a diet which included the Vitamin B6-antagonist, 4-DPD, both influence the course of IgG, IgG1 and IgM production.

Interleukin-1beta and interleukin-1alpha may affect the implantation rate of patients undergoing in vitro fertilization-embryo transfer.

OBJECTIVE: To investigate whether interleukin-1beta (IL-1beta) and interleukin-1alpha (IL-1alpha) affect the implantation rate of patients undergoing IVF-ET. DESIGN: Follicular fluid and serum were obtained on the day of hCG administration, the day of oocyte retrieval, and the day of embryo transfer. SETTING: Cellular immunology laboratory in a research institute, a high technology IVF unit in a medical center, and a university hospital. PATIENT(S): Thirty-three women who were undergoing IVF-ET. MAIN OUTCOME MEASURE(S): IL-1beta and IL-1alpha were measured by specific ELISA and their levels were correlated with the implantation rate. RESULT(S): Classification of IVF-ET patients according to their implantation rate revealed significantly higher amounts of follicular fluid IL-1beta in the implantation versus nonimplantation cycles (68.5+/-24.6 pg/mL versus 20.5+/-13.4 pg/mL); The difference between the level of IL-1alpha in the two groups was not statistically significant(11.6+/-5.1 pg/mL versus 7.3+/-1.9 pg/mL). In parallel, systemic FSH/hMG-dependent IL-1beta and IL-1alpha production was observed in implantation cycles but not in nonimplantation cycles. Statistically significant IL-1beta and IL-1alpha production was observed after administration of hCG. CONCLUSION(S): Gonadotropins used during IVF-ET induce local and systemic production of IL-1beta and IL-1alpha. In addition, the implantation rate for IVF-ET patients who have detectable serum concentrations of IL-1beta and IL-1beta on the day of hCG administration could be higher than the rate for IVF-ET patients who do not have detectable concentrations of these cytokines.

ISCOMs vaccine against experimental leishmaniasis.

The major surface glycoprotein (gp63) of Leishmania major incorporated into the immunostimulating complexes (ISCOMs) was used to protect Balb/c mice against experimental infection. Two intraperitoneal vaccinations with low doses of gp63 into ISCOMs (gp63-ISCOMs) induced protective immunity in vaccinated mice as indicated by reduced inflammation and suppressed lesions after experimental challenge. An augmented IgG-specific secretion and a specific switching towards the IgG2a isotype was observed in the serum of vaccinated mice. Gp63-ISCOMs primed spleen cells restimulated in vitro with soluble Leishmania antigen (SLA) or live parasites displayed strong gp63-specific proliferative responses and secreted high levels of interleukin-2, interferon gamma and interleukin-10 but not interleukin-4. No delayed type hypersensitivity response to either SLA or LV39 was detected. These data indicate that gp63-ISCOMs induced a protective immunity in the susceptible Balb/c mice against Leishmania challenge, modulating the immune response towards a Th1 rather than Th2 type.

Influence of Quillaja saponaria triterpenoid content on the immunomodulatory capacity of Epstein-Barr virus iscoms.

The immune responses to immunostimulating complexes (iscoms) containing recombinant Epstein-Barr virus (EBV) gp340 envelope protein was evaluated in BALB/c (H-2(d)) and CBA (H-2(k)) mice. Gp340-iscoms were used either with a low content of Quillaja triterpenoid adjuvant (L-iscoms) or supplemented with additional Quillaja adjuvant in the form of iscomatrix (S-iscoms). Class and subclass distribution of anti-gp340 antibodies, EBV-neutralizing antibodies, antigen-specific T cell proliferation and cytokine production were determined and these results compared to those obtained by immunization with non-adjuvated gp340. The H-2(d) and H-2(k) mice were characterized as low or high responders in respect to the level of specific anti-gp340 antibodies, secretion of IgG2a isotype, antigen-specific lymphoproliferative capacity, interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) production in the basic immunizations with gp340. While presentation of the antigen in iscom formulations with low levels of Quillaja triterpenoids induces a moderate enhancement of the immune responses in the low responder H-2(d) mice, supplementation with high levels of iscomatrix immunomodulator was required to enhance the immune responses in the high responder H-2(k) mice. In both mouse strains subcutaneous immunization with S-iscoms resulted in a significant increase of IgG1- and IgG2a-specific antibodies, as well as in strong antigen-specific proliferative response confirmed by the simultaneous cytokine production. The enhanced antigen-specific secretion of IL-2 and IFN-gamma together with the abrogation of IL-10 and the absence of IL-4 indicates that the responses were driven towards a Th1-type rather than Th2-type immune response. The S-iscom formulations minimized the differences in immune responses between the two mouse strains, but the capacity of immune sera to neutralize EBV transformation in vitro remained completely strain-dependent. These data indicate that immune responses generated by iscoms can be manipulated by altering the triterpenoid composition of the iscoms and that the levels of triterpenoids can determine whether or not a Th1-type response is made.

Correlation of serum and follicular fluid concentrations of placental protein 14 and CA-125 in in vitro fertilization-embryo transfer patients.

OBJECTIVE: To study serum and follicular fluid (FF) concentrations of placental protein 14 (PP14) and CA-125 in patients participating in an IVF-ET program. DESIGN: Determination in serum and FF on the day of hCG administration, day of oocyte retrieval, and on ET day of PP14 and CA-125. SETTING: Second Department of Obstetrics and Gynecology of the University of Athens; Euromedica IVF Unit; and Hellenic Pasteur Institute. PATIENTS: Thirty-three patients undergoing IVF-ET divided in three groups: 8 with conception, 18 with fertilized oocytes but no conception, and 7 without fertilized oocytes. INTERVENTIONS: All patients underwent an ovarian stimulation with a short protocol of GnRH analogue-pure-FSH-hMG. Three blood probes were collected from each patient, on the day of hCG administration, on retrieval day, and on ET day, respectively, whereas FF was collected on retrieval day. MAIN OUTCOME MEASURES: Determination in serum and FF of CA-125 and PP14. RESULTS: On the day of oocyte retrieval, both mean values of serum PP14 and CA-125 were significantly higher in conceptional than nonconceptional cycles. On the day of ET, the mean values of serum PP14 increased significantly in conceptional cycles whereas CA-125 showed no difference. The mean concentration of PP14 in FF was significantly higher in conceptional cycles whereas there was no significant change in the mean concentration of FF CA-125. CONCLUSIONS: In conceptional cycles after IVF, PP14 increases in serum significantly from hCG day to ET day compared with nonconceptional cycles, whereas CA-125 increases from hCG day to oocyte retrieval day and decreases on ET day. In FF, PP14 was increased in conceptional cycles, whereas CA-125 in FF showed no change.

Canine visceral leishmaniasis in the great Athens area, Greece.

A survey of 1,175 dogs with suspected visceral leishmaniasis (VL), was carried out to investigate canine leishmaniasis in the Athens basin, the largest metropolitan area of Greece. The dogs examined were not a random sample since animals were selected on the basis of symptoms indicating the disease. Dog sera were tested using the indirect fluorescent antibody technique (IFAT), with titres greater than or equal to 1/200 considered positive for VL. On this basis 569 dogs (48.4%) were found positive for VL and 69 (5.9%) borderline (with titres of 1/100). Additionally, 218 of these sera were also tested by enzyme-linked immunosorbent assay (ELISA). Of this sample, 120 were negative and 88 were positive with both IFAT and ELISA (according to the stated criteria), and thus the concordance of the two methods was 98.6%. Dogs positive for VL were classified according to age, sex, breed, and area of residence. Dogs from outlying areas of the Athens basin had higher infection rates than dogs from the city. The most affected breed was the Doberman, and the least affected was the Collie.

Generation of TNF alpha, IFN gamma, IL-6, IL-4 and IL-10 in mouse serum from trichinellosis: effect of the anti-inflammatory compound 4-deoxypyridoxine (4-DPD).

Infections caused by the nematode Trichinella spiralis is characterized in the host by an inflammatory response with cytokine production. In these studies we have detected TNF alpha, IL-6, IFN gamma, IL-4 and IL-10 in the serum of 10 mice infected with T. spiralis. Moreover, we detected, for the first time, these cytokines in the serum of mice treated with 4-DPD, a potent antagonist of vitamin B6 coenzyme which has anti-inflammatory properties. 4-DPD was used at 100, 400, 800 micrograms/bolus for 20 days, starting one day before the infection. After 15 days of T. spiralis infection, TNF alpha reached a maximum level, while IL-6 was maximal after 7 days, IFN gamma at 20 days and IL-4 at 14 days. IL-10 was not affected by the T. spiralis infection. When the animals were treated with 4-DPD at the reported dosages and infected with T. spiralis the inhibition of TNF alpha and IL-6, were dose-dependent in the first 7 days while IL-4 was reduced only at 400-800 micrograms/bolus. 4-DPD-treated mice did not statistically (P > 0.05) affect the generation of IFN gamma. In healthy animals the production of cytokines were not measurable, just as it was in non-infected animals treated with 4-DPD. The increase of cytokines such as, TNF alpha and IL-6 may be related to the severity of the disease, boosting the host's resistance to the pathogen and inhibiting parasite survival. In addition, the augmentation of IL-4 production enhances T and B cells and macrophage responses and may stimulate T-cell antibody-mediated response to the pathogen. 4-DPD, an inhibitor of IL-1 and inflammatory reactions, proved to be most effective on TNF alpha and IL-6, which are mainly produced by macrophages.

Immunoglobulin and cytokine profile in murine secondary hydatidosis.

We have investigated specific immune responses in BALB/c mice with experimentally induced secondary hydatidosis. Following intraperitoneal inoculation of brood capsules containing Echinococcus granulosus-protoscoleces, the course of the infection was followed for 513 days. The sera of the mice were screened for the presence of a number of cytokines, and for specific antibodies. During the first 129 days of infection, high levels of cytokines TNF alpha, IL-alpha, IFN gamma, IL-6, and IL-10 and specific IgG1 and IgG3 isotypes were detected, as compared to uninfected controls. The levels of IgM and IgG2a were slightly increased following infection, and remained elevated throughout the period of observation. The levels of IL-1 alpha and specific immunoglobulin of all isotypes except IgM and IgG2a, were significantly decreased 103 days post infection (p.i.), whereas TNF alpha was sharply decreased 129 days p.i. During the period of 129 to 209 days of infection there was an increase in secreted IL-10, and a slow decrease in the levels of IL-6 and IFN gamma. Levels of IgM, IgG, IgG1, and IgG2a plateaued during this period, whereas IgG3 and TNF alpha showed a peak at day 190 p.i. These data suggest the induction of Th2 antibody-mediated immunity with a parallel expansion of Th1-mediated inflammatory responses as important mechanism of host defence against the metacestode.