Effect of vitamin C in reducing the toxicity of endosulfan in liver in rabbits

In this study, the effect of endosulfan, an organochlorine pesticide, and the ameliorating effect of vitamin C on the livers of New Zealand white rabbits were studied. Livers of the rabbits were examined grossly and histopathologically, and caspase-3 activity was detected by immunohistochemical methods. A total of twenty-four rabbits were divided into four groups (n=6). Rabbits in Group I (END) were daily given a sublethal dose of endosulfan (1 mg/kg bw) in corn oil by oral gavage for 6 weeks. Group II (END+C) received the same dose of endosulfan and additionally Vit C (20 mg/kg bw) every other day during this period. Group III (OIL+C) received corn oil daily by oral gavage and vitamin C every other day for 6 weeks. Group IV (OIL), the control group, received only corn oil daily, by oral gavage throughout the experiment. The concentration of α-endosulfan in the END group was higher in livers (0.102±0.012 ppb) than the β-endosulfan (0.072±0.001 ppb). Decreased accumulation of α and β endosulfan was observed in the END+C group (0.025±0.003 and 0.016±0.002 ppb, respectively) (p<0.0001). The most prominent gross findings at the necropsy were seen in the END group, in which swollen and pale livers were commonly observed. Hemorrhages, degenerations, necrosis, and in some rabbits bile duct hyperplasia were the marked histopathological findings of the END group. Caspase-3 positive reaction was more severe in this group than in the others. An ameliorating effect of Vit C on gross, histopathological and immunohistochemical findings was observed in the END+C group. The results revealed that endosulfan is highly toxic for rabbit livers. However, toxicity was decreased by Vit C treatment, which reduced the accumulation of endosulfan in livers four-fold.     

Analysis of micronucleus induction of pyrimethamine in in vitro CHL cells and in in vivo mouse bone marrow cells

In vivo and in vitro mutagenicity of pyrimethamine were examinedin the micronucleus test. Pyrimethamine strongly induced micronucleiin a dose-dependent manner in the in vitro micronucleus testusing the Chinese hamster lung (CHL) cell line, when treatedat 0.2–1.6 µg/ml for 48 h. The in vivo micronucleustest was carried out in mice after the first, second, thirdand fourth administration of doses up to 40 mg/kg p.o. The resultsshowed no increased frequency of micronuclei after any treatment,though pyrimethamine was shown to persist at levels >2 µ/mlin plasma after a single oral administration of 50 mg/kg. ¹To whom correspondence should be addressed     

Role of Nitric Oxide in Prevention of Cognitive Disorders in Neurodegenerative Brain Injuries in Rats

NO synthesis disturbances play an important role in the development of neurodegenerative damage in Alzheimer disease. We previously showed that adaptation to intermittent hypobaric hypoxia prevents cognitive disturbances in rats with experimental Alzheimer disease [6]. Here we evaluated the role of NO in cognitive disorders and development of adaptive protection during experimental Alzheimer disease. Adaptation to hypoxia in rats was performed in a hypobaric pressure chamber at a simulated altitude of 4000 m (4 h per day for 14 days). Alzheimer disease was simulated by bilateral injections of a toxic fragment of β-amyloid (25–35) into n. basalis magnocellularis. For evaluation of the role of NO in the development and prevention of memory disorders, the rats received intraperitoneally either NO-synthase inhibitor N^ω-nitro-L-arginin (L-NNA, 20 mg/kg, every other day for 14 days) or NO-donor dinitrosyl iron complex (200 μg/kg daily for 14 days). NO-synthase inhibitor potentiated the damaging effect of β-amyloid, abolished the protective effect of adaptation to hypoxia, and produced memory disorders in rats similar to those observed during experimental Alzheimer disease. In contrast, the increase in NO level in the body provided by injections of the NO-donor produced a protective effect against memory disorders caused by β-amyloid similar to that induced by adaptation to hypoxia. We concluded that reduced NO production in the organism plays an important role in the development of cognitive disorders produced by injections of β-amyloid, while prevention of NO deficit by administration of NO-donors or nonpharmacological stimulation of NO synthesis can provide a protective effect in experimental Alzheimer disease. Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 146, No. 10, pp. 371–375, October, 2008     

Evaluation of roxithromycin in the treatment of non-gonococcal urethritis in males

One-hundred and fifty-two male patients suffering from non-gonococcal urethritis were treated with an oral dosage of 300 mg roxithromycin daily for seven days.Chlamydia trachomatis was isolated from the urethra in 53 patients (35 %), and Ureaplasma urealyticum in 42 patients (28 %). After treatment, 49 (92 %) of the 53 patients with positive Chlamydia trachomatis cultures and 34 (81 %) of the 42 patients with positive Ureaplasma urealyticum cultures had negative cultures at follow-up. A clinical cure was observed in 137 patients (90 %). Ten patients (7 %) showed side effects consisting of nausea, sensation of distended abdomen, headache and fatigue. Seventy-eight male patients suffering from nongonococcal urethritis were treated with an oral dosage of 2×150 mg roxithromycin daily for seven days.Chlamydia trachomatis was isolated from the urethra in 22 patients (28 %), and Ureaplasma urealyticum in 30 patients (38 %). After treatment, all of the 22 patients with formerly positive Chlamydia trachomatis cultures and 23 (77 %) of the 30 patients with formerly positive Ureaplasma urealyticum cultures were negative at follow-up. A clinical cure was observed in 70 patients (90 %). Three patients (4 %) showed side-effects consisting of nausea and headache. It is concluded that roxithromycin is a good alternative to tetracycline and erythromycin in the treatment of non-gonococcal urethritis in males.     

Demonstration of enzymes in macrophages cultivated in vitro and in epitheloid cells

Zusammenfassung An Makrophagen und epitheloiden Zellen aus Kulturen von Mäusegeweben, nämlich Milz, embryonaler Lunge und auch aus einem Spontantumor, lassen sich mit histochemischen Methoden folgende Enzyme auffinden: Adenosintriphosphatase, saure Phosphatase und unspezifische Esterase. Nachweisbare alkalische Phosphatase ist nicht vorhanden.Die Aktivität dieser Enzyme ist in den Makrophagen und epitheloiden Zellen deutlich stärker als z. B. in Fibroblasten. Die Adenosintriphosphatase ist in den histiocytären Zellformen, ohne eine feinere Lokalisation zuzulassen, vorwiegend in einer perinucleären Zone, bei den Makrophagen auch in den pseudopodienartigen Fortsätzen festzustellen, in den Fibroblasten und Carcinomzellen dagegen fast ausschließlich in den Mitochondrien. Die nachweisbare unspezifische Esterase ist im wesentlichen an der Zelloberfläche lokalisiert.Die reiche, histochemisch feststellbare Enzymausstattung der Makrophagen und epitheloiden Zellen in vitro wird auf den allgemeinen Anregungszustand dieser Zellen zurückgeführt, zu dem es bei der Umwandlung aus ruhenden histiocytären Zellformen unter dem Reiz der Explantation kommt.

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Summary Adenosintriphosphatase, acid phosphatase andnon-specific esterase have been demonstrated in cultivated macrophages and epitheloid cells from tissue cultures of mouse spleen, embryonic lung, and of a mouse mammary carcinoma. These enzymes give a considerably stronger reaction in macrophages and epithelioid cells than in fibroblasts. No alkaline phosphatase has been observed. Adenosintriphosphatase in macrophages and epithelioid cells is mainly found in a perinuclear zone but also in the pseudopodial cytoplasmic processes of macrophages; in fibroblasts and carcinoma cells, however, this enzyme gives a strongly positive reaction almost exclusively in mitochondria.Acid phosphatase shows a uniform distribution in the cytoplasm of all cells examined. Enzymatic staining of the cell surface was observed bynon-specific esterase reaction.It is suggested that the rich enzymatic system of cultivated macrophages and epithelioid cells — as far as it can be visualized by certain histochemical methods — may develop during the activation of resting histiocytic cells and their transformation ocurring under the stimulus of explantation.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Role of peptide-leukotrienes in liver injury in mice

The role of peptide leukotrienes (p-LTs), especially LTC₄ and LTD₄ in liver disease, was investigated in mice experimental liver injury models. The liver injury was induced by the injection of bacterial lipopolysaccharide (LPS) intoCorynebacterium parvum pretreated mice. Carbon tetrachloride (CCl₄)-induced liver injury in mice was used as a standard model. In both injury models, extensive liver parenchymal cell damage was observed by the elevation of glutamate transaminase (GOT and GPT) activity and confirmed by significant histopathological changes in the liver. Moreover, significant elevation of LTC₄ in the liver was observed in both models 1 and 6 h after the onset of disease. Administration of AA-861, a selective 5-lipoxy-genase inhibitor (0.5, 1, and 2 mg/kg) and LY-171883, a p-LT receptor antagonist (50 and 200 mg/kg) suppressed the elevation of serum GOT and GPT levels and histopathological changes in both experimental liver injury models. In addition, when authentic LTC₄ or LTD₄ was injected into the mouse, clear elevation of serum GOT and GPT and histopathological changes of the liver were observed. These results suggest that p-LTs play a role in the onset of liver diseases in mice.     

The role of immunoconglutinin in the in vivo and in vitro destruction of red cells

Two experimental approaches were used to investigate the role of immunoconglutinin (IK) in the in vivo destruction of red cells in rabbits. In a first series of experiments the behaviour of EC43 (C3-coated red cells) was followed in IK-producing rabbits and in rabbits passively receiving IK, both of which had previously been complement-depleted by cobra venom factor (CoF). The red cells were sequestered to a minor degree and returned to the circulation within 20–30 min, whereas in the normal control, EC43 returned to the circulation over a period of 3–4 hr. In contrast to EC43, EC43IK (IK-coated EC43) did not form rosettes around the Kupffer cells, suggesting that IK blocks the functional activity of C3 and so interferes with the interaction between C3 and C3-receptors on fixed macrophages. In a second series of experiments EC43 and EC43IK, injected into IK-producing rabbits and normal rabbits respectively, underwent marked lysis and erythrophagocytosis. Examination of liver imprint preparations from these rabbits revealed rosette formation around the Kupffer cells, indicating the fixation of more C4 and C3 by bound IK.

In vitro experiments confirmed both the inhibitory activity and the complement-fixing capacity of IK.

The results suggest that IK normally has an amplifying effect on complement fixation in vivo and so potentiates the ability of complement to bring about red-cell destruction.

     

Prevalence of coccidia in beef cattle in western Turkey

A total of 504 bovine faecal samples collected from intensively managed beef farms in Afyon province of Turkey were examined to determine the types and prevalence of coccidian parasites present. Coccidian oocysts were found in 20.04% of all the samples examined by sucrose-flotation. The species detected and their prevalence were Eimeria bovis (34.55%), E. auburnensis (23.03%), E. canadensis (14.55%), E. brasiliensis (10.91%), E. zuernii (6.67%), E. bukidnonensis (3.03%), E. cylindrica (2.42%), E. ellipsoidalis (1.21%), E. illinoisensis (1.21%), E. alabamensis (1.21%) and Isospora sp. (1.21%). Mixed infections of two to four species were found in 43.6% of the animals. The overall prevalence of coccidial oocysts in faecal samples was 27.23% for calves, 15.65% for cows. Linear regression analysis showed that there is a significant reduction in the OPG (the number of oocysts per gram of feces) levels (P < 0.05) in cows infected with Eimeria. No cases of clinical coccidiosis were observed in this survey.     

Molecular Characterization of Bacillus Strains Involved in Outbreaks of Anthrax in France in 1997

Outbreaks of anthrax zoonose occurred in two regions of France in 1997. Ninety-four animals died, and there were three nonfatal cases in humans. The diagnosis of anthrax was rapidly confirmed by bacteriological and molecular biological methods. The strains of Bacillus anthracis in animal and soil samples were identified by a multiplex PCR assay. They all belonged to the variable-number tandem repeat (VNTR) group (VNTR)₃. A penicillin-resistant strain was detected. Nonvirulent bacilli related to B. anthracis, of all VNTR types, were also found in the soil.     

The efficacy of ribavirin in the treatment of Crimean-Congo hemorrhagic fever in Eastern Black Sea region in Turkey

BackgroundThe efficiency of ribavirin for treatment of Crimean-Congo hemorrhagic fever (CCHF) is unknown. In the literature, prospective randomized studies investigating the efficacy of ribavirin are not found.ObjectivesTo investigate the efficacy of ribavirin in treatment of patients with CCHF.Study designIn this prospective randomized cohort study 136 cases were included between June 2004 and August 2007. The diagnosis was confirmed in the CCHF reference laboratory of Refik Saydam National Hygiene Central Institute of the Turkish Ministry of Health. Patients either received ribavirin plus supportive treatment (Group A) (n = 64) or only supportive treatment (Group B) (n = 72). For the evaluation of efficacy of ribavirin, various parameters were compared between Group A and Group B.ResultsAs well as the similarity of demographic features between the two groups, there were no statistical differences in incubation time; hospitalization time; patients requiring platelet replacement therapy; the time taken for platelet levels to return to normal levels and mortality. In Group B, the rate of tick contact was higher (p = 0.03). In Group A, leukocyte levels took longer to return to the normal levels (p = 0.02).ConclusionIn our study, there was no positive effect determined on clinical or laboratory parameters in CCHF patients treated with ribavirin, also it was observed that leukocyte levels took longer to return to normal (p = 0.02) and, while not statistically significant, the longer period of hospitalization (p = 0.09) needed was observed as a negative effect. Because of these reasons, it is thought that the use of ribavirin makes no significant contribution to the prognosis of the CCHF disease.     

Role of the cytoskeleton in stimulation of Na+ channels in A6 cells by changes in osmolality

 Permeable supports with A6 cell monolayers were mounted in an Ussing chamber and bilaterally bathed with Ringer solution at room temperature. Short-circuit current (I _sc) was recorded continuously, and noise analysis revealed microscopic channel current characteristics. Our investigation focuses on the stimulation of apical Na⁺ entry caused by exposing the serosal surface of the A6 cell monolayers to hyposmotic Ringer solution. To evaluate the possible role of the cytoskeleton in the regulation of Na⁺ channels in response to a change in osmolality we used four different experimental approaches. In the control group, which were not exposed to any cytoskeleton-influencing drugs, there was a 1.5-fold increase in I _sc and in the number of open Na⁺ channels after osmotic stimulation. For the second group cytochalasin D (0.1 μg/ml) was present on the serosal side during the experiments. Neither I _sc nor the number of open Na⁺ channels increased after osmotic stimulation. In the third group colchicine (0.2 mM) or nocodazole (20 μM) was present on the serosal side, which resulted in 1.8-fold and 1.5-fold increases in I _sc as well as 3-fold and 2-fold increases in the number of Na⁺ channels, respectively. In the fourth experimental group erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (EHNA, 0.5 mM), a dynein inhibitor, was present on the serosal side. In this group I _sc decreased to about 0.4 μA/cm², and subsequent application of amiloride abolished I _sc completely. Under hyposmolar conditions EHNA abolished entirely the sensitivity of I _sc to the osmotic challenge. Because of the EHNA-induced down-regulation of I _sc, the density of apical Na⁺ channels in this experimental group could not be determined. These results show that the cytoskeleton is dominantly involved in osmotic channel regulation at the apical membrane, and that actin filaments, microtubules and molecular motors are involved in the recruitment of additional Na⁺ channels. Received: 21 July 1997 / Received after revision: 4 December 1997 / Accepted: 16 December 1997     

Genetic predictors of inflammation in the risk of occupational asthma in young apprentices

BackgroundThe influence of genetic predictors of inflammation and atopy on occupational asthma in apprentices is not known.ObjectivesTo assess the influence of genetic polymorphisms of IL4RA, IL13, TNFA, IL1A, and IL5 on the decline of lung function and bronchial hyperresponsiveness in a prospective follow-up study of baker/pastry maker and hairdresser apprentices.MethodsA total of 351 apprentices were included in the study. We performed skin testing, spirometry, fractional exhaled nitric oxide measurement, and methacholine hyperreactivity testing at the initial visit and during and at the end of the 18-month training period. Gene variants of IL4RA, IL13, TNFA, IL1A, and IL5 were determined in DNA from nasal lavage.ResultsIL13 R130Q/IL4RA S478P or IL13 R130Q//IL4RA Q551R were significant predictors of the decrease of forced expiratory volume and forced vital capacity (P ≤ .006). Genotype GG of TNFAG308A was associated with bronchial hyperresponsiveness in the whole population and in nonatopic individuals (90.63% vs 9.38%; odds ratio, 3.78; 95% confidence interval, 1.10-12.83). TNFA GA and IL5 CC and TNFA GA and IL1A CC were 2 epistatic predictors of exhaled nitrogen monoxide decrease during follow-up (P = .02 and P = .004, respectively). The association with TNFA GA and IL1A CC was the most significant in nonatopic bakers (P < .001).ConclusionWe evidenced a predicting influence of IL13/IL4RA and TNFA in the early exposure to allergens and irritants that precedes occupational asthma. The significance of the associations in the absence of atopy suggests an influence of the genetics predictors related to inflammatory pathways.     

Inhibitory role of Smad7 in hepatocarcinogenesis in mice and in vitro

Smad7 is a principal inhibitor of the TGFβ–Smad signalling pathway. We have investigated the functional significance of Smad7 in hepatocellular carcinoma (HCC). Smad7 knockout (KO) and wild‐type (WT) mice were injected with diethylnitrosamine (DEN) to induce HCC. The effects of Smad7 on cellular features were examined in HCC cells, using a Smad7 over‐expression or deletion approach. Signalling pathway components modulated by Smad7 in HCC were evaluated using luciferase reporter assay and co‐immunoprecipitation. Smad7 was down‐regulated in human HCCs compared with the adjacent normal tissues (p < 0.001). Smad7 KO mice were more susceptible to DEN‐induced HCC than WT mice (78% versus 22%, p < 0.05). HCCs from KO mice displayed a greater proliferation activity (p < 0.05) and a reduced apoptotic index compared with WT littermates (p < 0.05). Deletion of Smad7 promoted cell proliferation in primary cultured HCC cells. In addition, over‐expression of Smad7 in HCC cell lines markedly suppressed cell growth (p < 0.0001) and colony formation (p < 0.01). Cell cycle analysis revealed an increase in the G₁ phase and a reduction in the S‐phase populations, accompanied by up‐regulation of p27^Kip1 and down‐regulation of cyclin D1. Smad7 increased cell apoptosis (p < 0.01) by mediating an intrinsic [caspase‐9, caspase‐3 and poly(ADP‐ribose) polymerase] apoptotic pathway. Moreover, Smad7 inhibited NF‐κB signalling by interacting with TAB2, an upstream activator of NF‐κB, and inhibited TGFβ signalling by suppressing phosphorylation of Smad3. In conclusion, loss of Smad7 enhances susceptibility to HCC. Smad7 suppresses HCC cell growth by inhibiting proliferation and G₁–S phase transition and inducing apoptosis through attenuation of NF‐κB and TGFβ signalling. Smad7 acts as a potential tumour suppressor in liver. Copyright © 2013 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

A Critical Role of Activin A in Maturation of Mouse Peritoneal Macrophages in vitro and in vivo

Activin A, a multifunctional factor of the transforming growth factor-beta （TGF-β） superfamily, is mainly produced by microglia and macrophages, and its anti-inflammatory and pro-inflammatory activities are both related to macrophage functions. However the direct effect of activin A on the rest macrophages in vivo remains unclear. In the present study, the results showed that activin A not only increased NO and IL-1β release, but also promoted phagocytic abilities of mouse peritoneal macrophages in vitro and in vivo, whereas it did not influence MHC Ⅰ and MHC Ⅱ expression. Moreover, we found that activin A significantly upregulated the expressions of CD14 and CD68, markers of mature macrophages, on the surface of macrophages in vitro and in vivo. These data suggest that activin A can induce primary macrophage maturation in vitro and in vivo, but may not trigger the acquired immune response via regulating expression of MHC molecules involved in presentation of antigen.     

Role of naso-buccal passages in thermoregulation in sheep

1. Experiments with a face-mask in which the temperature of the air in the face-mask was raised to 40° C while the ambient temperature in the chamber was maintained at 20° C, resulted in a marked increase in respiratory frequency and a slight decline in carotid blood temperature of unshorn sheep. Partially shorn sheep showed only small respiratory responses.

2. Localized infra-red irradiation of the naso-buccal area of unshorn sheep also resulted in an increased respiratory rate.

3. It is suggested that the initiation of polypnoea during infra-red irradiation of the naso-buccal region and following rise in the temperature of the air in the face-mask is due to stimulation of warm receptors in the upper respiratory tract.

4. Cooling the naso-buccal air in the face-mask to 10° C after thermal polypnoea had been established at an ambient temperature of 40° C resulted in a moderate decline of 30-40 respirations/min. This decline was attributed to the stimulation of cold receptors located in the upper respiratory tract.

     

Further antinociceptive effects of myricitrin in chemical models of overt nociception in mice

The present work explored the antinociceptive effects of the flavonoid myricitrin in models of overt nociception triggered by intraplantar injection of chemical algogens into the hind paw of mice. The nociception induced by bradykinin (3 nmol/paw i.pl.) was abolished by prior treatment with myricitrin (10–100 mg/kg, i.p.) with ID₅₀ of 12.4 (8.5–18.1) mg/kg. In sharp contrast, myricitrin failed to affect the nociception elicited by prostaglandin E₂ (3 nmol/paw i.pl.). Cinnamaldehyde (10 nmol/paw i.pl.)-induced nociception was reduced by myricitrin (100 mg/kg, i.p.) and camphor (7.6 mg/kg, s.c.) in 43 ± 10% and 57 ± 8%, respectively. Myricitrin (30–100 mg/kg, i.p.) and amiloride (100 mg/kg, i.p.) inhibited nociceptive responses induced by acidified saline (pH 5/paw i.pl.), with ID₅₀ of 22.0 (16.1–30.0) mg/kg and inhibition of 71 ± 6% and 64 ± 5%, respectively. Moreover, myricitrin (10–30 mg/kg, i.p.) and ruthenium red (3 mg/kg, i.p.) significantly reduced the nociception induced by menthol (1.2 μmol/paw i.pl.) with the mean ID₅₀ of 2.4 (1.5–3.7) mg/kg and inhibition of 95 ± 3% and 51 ± 7%, respectively. In addition, myricitrin administration (30 and 100 mg/kg, i.p.) markedly reduced menthol-induced mechanical allodynia. However, myricitrin (100 mg/kg, i.p.) prevented (only in time of 60 min) cold allodynia induced by menthol. Collectively, the present results extend prior data and show that myricitrin promotes potent antinociception, an action that is likely mediated by an inhibition of the activation of nociceptors by bradykinin and TRPs agonist (i.e. cinnamaldehyde, acidified saline and menthol), probably via inhibition of PKC pathways. Thus, myricitrin could constitute an attractive molecule of interest for the development of new analgesic drugs.     

Evaluation of the effects of ozone therapy in the treatment of intra-abdominal infection in rats

INTRODUCTION

The antibacterial effect of ozone (O₃) has been described in the extant literature, but the role of O₃ therapy in the treatment of certain types of infection remains controversial.

OBJECTIVES

To evaluate the effect of intraperitoneal (i.p.) O₃ application in a cecal ligation/puncture rat model on interleukins (IL-6, IL-10) and cytokine-induced neutrophil chemoattractant (CINC)-1 serum levels, acute lung injury and survival rates.

METHODS

Four animal groups were used for the study: a) the SHAM group underwent laparotomy; b) the cecal ligation/puncture group underwent cecal ligation/puncture procedures; and c) the CLP+O₂ and CLP+O₃ groups underwent CLP+ corresponding gas mixture infusions (i.p.) throughout the observation period. IL-6, CINC-1 and IL-10 concentrations were determined by enzyme-linked immunosorbent assay (ELISA). Acute lung injury was evaluated with the Evans blue dye lung leakage method and by lung histology. P<0.05 was considered significant.

RESULTS

CINC-1 was at the lowest level in the SHAM group and was lower for the CLP+O₃ group vs. the CLP+O₂ group and the cecal ligation/puncture group. IL-10 was lower for the SHAM group vs. the other three groups, which were similar compared to each other. IL-6 was lower for the SHAM group vs. all other groups, was lower for the CLP+O₃ or CLP+O₂ group vs. the cecal ligation/puncture group, and was similar for the CLP+O₃ group vs. the CLP+O₂ group. The lung histology score was lower for the SHAM group vs. the other groups. The Evans blue dye result was lower for the CLP+O₃ group vs. the CLP+O₂ group and the cecal ligation/puncture group but similar to that of the SHAM group. The survival rate for the CLP+O₃ group was lower than for the SHAM group and similar to that for the other 2 groups (CLP and CLP+O₂).

CONCLUSION

Ozone therapy modulated the inflammatory response and acute lung injury in the cecal ligation/puncture infection model in rats, although there was no improvement on survival rates.     

Diversity of Babesia in Ixodes ricinus ticks in Poland

PurposeThe aims of this study were: (1) to estimate Babesia prevalence in the most common species of tick in Poland, Ixodes ricinus, in two recreational areas (Urwita?t in the Mazury Lake District and Bielański Forest in Warsaw), and (2) to evaluate the molecular diversity of Babesia isolates in questing I. ricinus in Poland.Material and MethodsQuesting ticks were collected from vegetation in forest areas in Urwita?t near Miko?ajki and in Bielański Forest (Warsaw). Purified genomic DNA was used with specific primers to amplify a fragment of the Babesia spp. 18S rRNA gene.ResultsTick-drag indices for I. ricinus were high in both study areas, reaching somewhat higher values in Urwita?t than in Bielański Forest. The overall prevalence of Babesia spp. in examined ticks was 1.6%. In Urwita?t, two strains of B. microti were identified using rRNA sequences: the enzootic Munich strain and an isolate close to the zoonotic Jena strain. The proportion of infections due to these two strains in questing ticks reversed over a six-year period. During 3 years of study in Bielański Forest, all Babesia isolates obtained from I. ricinus were identical to Babesia sp. EU1 (B. venatorum), previously recognized as an agent of human babesiosis.ConclusionsThis study has confirmed the presence of enzoonotic and zoonotic Babesia species/strains in the abundant human-biting tick I. ricinus in recreational areas in Poland. It has also shown that the distribution of different genotypes has changed over time, however the reasons for these fluctuations still remain to be investigated.