Celecoxib treatment dampens LPS-induced periapical bone resorption in a mouse model.

AIM: To evaluate the efficacy of selective and nonselective inhibitors of cyclooxygenase-2 enzymes in the treatment of experimental apical periodontitis induced by bacterial lipopolysaccharide (LPS) in vivo in a mouse model. METHODOLOGY: Thirty-six C57BL/6 mice were used. After access cavity preparation, a solution containing E. coli LPS (1.0 µg µL-1 ) was inoculated into the root canals of the mandibular and maxillary right first molars (n = 72) After 30 days, apical periodontitis was established and the animals were systemically treated with celecoxib, a selective COX-2 inhibitor (15 mg kg-1 ), or indomethacin, a nonselective COX-2 inhibitor (5 mg kg-1 ), for 7 and 14 days. Blocks containing teeth and bone were removed for histopathological and histometric analyses (haematoxylin and eosin), evaluation of osteoclasts numbers (tartrate-resistant acid phosphatase enzyme - TRAP) and immunohistochemistry for RANK, RANKL and OPG. Gene expression was performed using reverse transcription and real-time polymerase chain reaction (qRT-PCR) for RANK, RANKL, OPG, TRAP, MMP-9, cathepsin K and calcitonin receptor. Histopathological, histometric, TRAP, immunohistochemistry and qRT-PCR data were evaluated using Kruskal-Wallis followed by Dunn's test (α = 0.05). RESULTS: Systemic administration of celecoxib for 7 and 14 days prevented periapical bone resorption (P < 0.0001), differently from indomethacin that exacerbated bone resorption at 7 days (P < 0.0001) or exerted no effect at 14 days (P = 0.8488). Celecoxib treatment reduced osteoclast formation in apical periodontitis, regardless of the period of treatment (P < 0.0001 for 7 days and P = 0.026 for 14 days). Administration of celecoxib or indomethacin differentially modulated the expression of genes involved in bone resorption. At 7 days, celecoxib and indomethacin treatment significantly inhibited expression of mRNA for cathepsin K (P = 0.0005 and P = 0.016, respectively) without changing TRAP, MMP-9 and calcitonin receptor gene expression. At 14 days, celecoxib significantly inhibited expression of mRNA for MMP-9 (P < 0.0001) and calcitonin receptor (P = 0.004), whilst indomethacin exerted no effect on MMP-9 (P = 0.216) and calcitonin receptor (P = 0.971) but significantly augmented cathepsin K gene expression (P = 0.001). CONCLUSIONS: The selective COX-2 inhibitor celecoxib reduced osteoclastogenic signalling and activity that dampened bone resorption in LPS-induced apical periodontitis in mice, with greater efficacy than the nonselective inhibitor indomethacin.

Pre-ozonation in the activated sludge process: fate of nitrogen species.

The objective of this research was to include ozonation prior to an activated sludge treatment and investigate the effect on the nitrogen species, their fate and the consequences of this oxidation upon the biomass. Three parallel treatment systems were used: the base system, where feed went directly to the activated sludge reactor, and two others, where the influent was ozonated at two different dosages, 15 and 25 mg/L of influent, prior to the biological reactors. The results from the ozonation chamber show a high oxidation capacity of the entering ammonia and organic nitrogen, proportional to the ozone dose. The oxidation product was nitrate. No de-nitrification was expected because a high oxygen concentration (4 mg/L) was maintained in the reactors. The reactors receiving ozonated influent showed a lower assimilation of nitrogen by the biomass. The sludge nitrogen content resulted in 11, 9.3 and 7.4% dry-weight corresponding to no-ozone, low ozone and high ozone dosages, respectively. In spite of the lower ammonia available in the ozonated flows, the corresponding reactors showed a higher specific nitrification rate. The ozonated system also performed better in terms of chemical oxygen demand (COD) and biochemical oxygen demand (BOD5) removals, besides showing a higher true biomass yield coefficient.

A retrospective study of mandibular fracture in a 40-month period.

This retrospective study evaluated the epidemiology, treatment and complications of mandibular fracture associated, or not associated, with other facial fractures, when the influence of the surgeon's skill and preference for any rigid internal fixation (RIF) system devices was minimized. The files of 700 patients with facial trauma were available, and 126 files were chosen for review. Data were collected regarding gender, age, race, date of trauma, date of surgery, addictions, etiology, signs and symptoms, fracture area, complications, treatment performed, date of hospital discharge, and medication. 126 patients suffered mandibular fractures associated, or not, with other maxillofacial fractures, and a total of 201 mandibular fractures were found. The incidence of mandibular fractures was more prevalent in males, in Caucasians and during the third decade of life. The most common site was the condyle, followed by the mandibular body. The therapy applied was effective in handling this type of fracture and the success rates were comparable with other published data.

Solitary and multiple erythema migrans in children: comparison of demographic, clinical and laboratory findings.

BACKGROUND: Data on European children with erythema migrans (EM) are limited. PATIENTS AND METHODS: 553 patients, 333 with solitary and 220 with multiple EM, diagnosed between 1996 and 2000, were included in the prospective study. Demographic, clinical and laboratory data including borrelial serum immunofluorescence assay antibody titers and Borrelia burgdorferi sensu lato blood culture results were obtained; findings in solitary and multiple EM were compared. RESULTS: Comparison revealed that children with multiple EM were younger (4.5 vs 6.5 years; p = 0.0000), less often reported a tick bite at the site of later skin lesion (25% vs 46%; p = 0.0000), had a longer incubation period (22 vs 13 days; p = 0.0028), more frequently presented with a ringlike lesion (99% vs 86%; p = 0.0000), less often reported associated local (15% vs 41%; p = 0.0000) but not systemic symptoms (28% vs 26%, p = 0.6913), more frequently had abnormal findings on physical examination (35% vs 26%; p = 0.0264), and a higher frequency of laboratory abnormalities including the presence of borrelial serum antibodies as well as B. burgdorferi sensu lato isolated from blood (12% vs 6%; p = 0.0267); younger age and male sex were identified as risk factors for the isolation of Borrelia. 40/44 isolates were Borrelia afzelii. CONCLUSION: Analysis of a large group of European children with solitary and multiple EM revealed several demographic, clinical and laboratory differences between the two groups.

First European pediatric case of human granulocytic ehrlichiosis.

Herein we report on the first confirmed pediatric case of acute human granulocytic ehrlichiosis in Europe. Presentation in this 11-year-old girl was comparable to clinical findings seen in adult European patients with human granulocytic ehrlichiosis; i.e., she had self-limited febrile illness with leukopenia, thrombocytopenia, and elevated serum C-reactive protein concentration. It is of interest that the patient not only had a fourfold change in antibody titer to Ehrlichia phagocytophila but also developed antibodies to Ehrlichia chaffeensis and that her PCR test result was positive on the third as well as on the 22nd day after the onset of illness, that is, 16 days after spontaneous defervescence.

Genotypic and phenotypic characterisation of Borrelia burgdorferi sensu lato strains isolated from human blood.

Lyme borreliosis often presents initially with erythema migrans. Borreliae may disseminate from the primary skin lesion, and different organs and systems could be affected. Borrelia strains were isolated from blood of 70 patients with Lyme borreliosis, including 10 patients from whom borreliae were also isolated from skin. The aim of the present study was to characterise the isolates with regard to their phenotypic and genotypic characteristics. Borreliae were cultivated in MKP medium. Species identification and plasmid profiles were determined by pulsed-field gel electrophoresis (PFGE) and protein profiles by SDS-PAGE. Digestion of Borrelia burgdorferi sensu lato DNA showed 63 (90%) B. afzelii Mla1 and 7 (10%) B. garinii Mlg2. No B. burgdorferi sensu stricto were isolated. Borreliae were isolated from both skin and blood of 10 patients, nine pairs of isolates were identical: seven B. afzelii and two B. garinii. B. afzelii was isolated from the skin and B. garinii from blood of the tenth patient. All but one isolate possessed at least one large plasmid and varying numbers of smaller plasmids. Eight (11.4%) of 70 isolates possessed an unusual plasmid profile (2 of 63 B. afzelii and 6 of 7 B. garinii). Borreliae differed in their protein profiles. OspA and OspB proteins were expressed by all B. afzelii isolates; 85.7% of B. garinii isolates expressed OspA and 71.4% expressed OspB. OspC was expressed by 65% of B. afzelii isolates and all B. garinii isolates. The ratios of B. afzelii and B. garinii isolated from blood and skin were similar. These results do not support the hypothesis that B. garinii has a higher propensity for haematogenous dissemination than B. afzelii. Antigen diversity as well as species and plasmid heterogeneity could play a role in the pathogenesis of the infection, suggesting distinctive strain organotropism.

Isolation of Borrelia burgdorferi sensu lato from blood of children with solitary erythema migrans.

OBJECTIVES: To establish the frequency of isolation of Borrelia burgdorferi sensu lato from blood of children with solitary erythema migrans (EM) in Europe, to determine the strains of the isolated borreliae and to compare the clinical course and the outcome of the disease according to positive and negative blood culture result. METHODS: In the prospective study we included 134 consecutive patients younger than 15 years with solitary EM, referred to our institution in 1996 and 1997. One milliliter of blood was withdrawn before treatment and cultured in modified Kelly-Pettenkofer medium. Isolated borreliae were typed according to LRFP analysis. Patients were treated with either penicillin V or cefuroxime axetil for 14 days. The posttreatment course was surveyed by follow-up visits during 1 year. RESULTS: B. burgdorferi sensu lato was isolated in 12 of 134 (9%) patients. Eleven blood isolates were typed: 10 were found to be B. afzelii and 1 was Borrelia garinii. Comparison of blood culture-positive and -negative patients revealed no differences in pretreatment characteristics or in posttreatment clinical course. However, worsening of local and/or systemic signs and symptoms at the beginning of antibiotic therapy (Jarish-Herxheimer's reaction) was identified more often in the blood culture-positive than in the blood culture-negative group (5 of 12 vs. 17 of 122, respectively; P = 0.0274). CONCLUSIONS: The isolation rate of B. burgdorferi sensu lato from the blood of children with solitary EM was 9%. The majority of the isolates were B. afzelii. Blood culture-positive patients treated with oral antibiotics were not at greater risk for unfavorable course of the disease than patients with negative blood culture result.

Demographic features, clinical characteristics and laboratory findings in children with multiple erythema migrans in Slovenia.

OBJECTIVE: To identify demographic, clinical and laboratory characteristics of children with multiple erythema migrans (EM) in Slovenia. METHODS: We prospectively studied patients aged 15 years and less, examined at our department for multiple EM in 1996 and 1997. Demographic and clinical data were collected by means of a questionnaire. In addition, basic haematological and biochemical investigations, serologic testing, and Borrelia cerebrospinal fluid and blood cultures were performed. RESULTS: Ninety-five children (44 girls, 55 boys) aged 1 to 13.5 (median, 4.5) years fulfilled the inclusion criteria. A tick bite was recalled by 23%. The incubation period was 10.5 (range, 1 to 150) days, the duration of skin lesions before the initial examination 4 (range, 1 to 54) days, and the median number of skin lesions, 4.5 (range, 2 to 35). The initial disease was mild in 81% of patients. Local and systemic symptoms were reported by 11% and 30% of children, respectively. Clinical signs accompanying EM lesions were found in 42%. Cerebrospinal pleocytosis (predominantly lymphocytic) was seen in 18% of patients; none of them had frank clinical evidence of central nervous system involvement. Serum IgM and IgG antibodies were detected in 28% and 22% of children, respectively. In 3/79 (4%) patients, Borreliae were isolated from the blood and in 2/83 (2%) from the cerebrospinal fluid. In 2/81 (2%) children, borrelial IgG intrathecal antibody production was demonstrated. CONCLUSIONS: Multiple EM in Slovene children is a mild disease. However, some patients had an associated, usually asymptomatic, infection of the central nervous system.

Intrathecal synthesis of specific antibodies in patients with invasion of the central nervous system by Mycoplasma pneumoniae.

Mycoplasma pneumoniae commonly causes respiratory tract infections in humans, but it may also be associated with central nervous system manifestations. The aim of the present study was to determine whether the cerebrospinal fluid taken from patients with neurologic symptoms due to Mycoplasma pneumoniae infection contains specific antibodies and whether the detection of these antibodies can be used for diagnosis. Mycoplasma pneumoniae was isolated from the cerebrospinal fluid taken from nine patients with central nervous system symptoms on admission to the hospital. In addition, Mycoplasma pneumoniae was detected in cerebrospinal fluid using polymerase chain reaction in four other patients. Antibodies to Mycoplasma pneumoniae were detected using the enzyme immunosorbent assay, indirect immunoperoxidase assay and immunoblotting in cerebrospinal fluid samples from 14 of 19 patients included in the study. The indirect immunoperoxidase assay showed high titers of Mycoplasma pneumoniae immunoglobulin G1 (IgG1) and IgM antibodies in cerebrospinal fluid samples of some patients with meningoencephalitis or meningitis. Titers of specific IgA, IgG2 and IgG3 antibodies were lower, while specific IgG4 was not detectable. Cerebrospinal fluid samples with higher antibody titers also contained IgA, IgG1, IgG2, IgG3 and IgM antibodies that recognized the P1 adhesin (170 kDa protein) of Mycoplasma pneumoniae. A comparison of antibody titers of concomitant serum/cerebrospinal fluid samples to Mycoplasma pneumoniae and those to measles virus by enzyme immunosorbent assay suggested the intrathecal synthesis of IgG and IgM antibodies to Mycoplasma pneumoniae in patients with acute meningoencephalitis. Data from this study clearly reinforce previous findings that Mycoplasma pneumoniae is an etiologic agent of central nervous system infections in humans.

Comparison of the epidemiological and clinical features of tick-borne encephalitis in children and adults.

The aim of this prospective study was to compare epidemiological data and clinical features in children and adults with tick-borne encephalitis (TBE). Patients with aseptic meningitis diagnosed at the University Medical Centre, Department of Infectious Diseases, Ljubljana, Slovenia, from June to August 1997, in whom the diagnosis of TBE was ascertained by the presence of serum IgM antibodies against TBE virus, who were serologically negative for Borrelia burgdorferi sensu lato and had a negative PCR CSF result on enteroviral infection, were included in the study. Out of 213 patients with aseptic meningitis, 80 (37.56%) fulfilled inclusion criteria. There were 20 children and 60 adults. In both groups males predominated. Virtually all patients had headache and fever, and more than 50% suffered from vomiting. The majority of patients in both groups recalled a tick bite, had a biphasic course of the illness, and was found to have obviously expressed meningeal signs. In both groups the median CSF leukocyte count was somewhat lower than 100 x 10(6)/l with a predominance of lymphocytes. Children were more often given antibiotics during the initial phase of TBE than adults (p = 0.0095). Several other statistically significant distinctions (p < 0.05) were found including the frequency of fatigue, malaise, vertigo, photophobia, myalgias, arthralgias, as well as elevated CSF albumin and protein concentration, elevated albumin quotient and IgG quotient; all these findings were more often present in adults. In addition a longer duration of fever, more frequent need for anti-edematous treatment and longer hospitalization were found in adults. Direct comparison of clinical and epidemiological characteristics of TBE in children and adults revealed differences in several clinical and laboratory features and corroborates the previous conclusion that TBE in childhood is a milder illness than TBE in adults.

Severe forms of tick-borne encephalitis in children.

Records of patients aged 0 to 15 years, hospitalised between 1993 and 1998 at the intensive care unit (ICU) of the Department of Infectious Diseases, Ljubljana, Slovenia, suffering from a severe attack of tick-borne encephalitis (TBE), were reviewed. Of 133 children hospitalised due to TBE virus infection during the observation period, 7 (5.2%) were treated in the ICU. All patients were male, aged 6 to 14 (mean, 11.1) years. In six cases, focal encephalitis was diagnosed, and in one case it was suspected. All patients survived. After a mean follow-up period of 7.9 (range, 1.5 to 17) months, one patient was found to have severe neurologic sequelae and two patients had moderate sequelae. In conclusion, the results of our retrospective study of severe forms of TBE in children demonstrate that this disease can run a severe course and may lead to permanent sequelae, most often in boys of school-going age who present with focal encephalitis. Therefore, immunisation of school children against TBE in endemic areas is strongly recommended.

Comparison of cefuroxime axetil and phenoxymethyl penicillin for the treatment of children with solitary erythema migrans.

OBJECTIVE: To compare the clinical efficacy and drug-related adverse effects of 14 days of treatment with cefuroxime axetil 30 mg/kg/day or phenoxymethyl penicillin 100,000 IU/kg/day in the treatment of children with erythema migrans. METHODS: Consecutive patients younger than 15 years, referred to our institution in 1996 with solitary erythema migrans and without prior antibiotic therapy, were included in this prospective study. Basic demographic features and clinical data were collected by questionnaire. The efficacy of the treatment of acute disease, development of major and/or minor manifestations of Lyme borreliosis and drug-related adverse effects were surveyed at follow-up visits during the first year after the initiation of antibiotic treatment. RESULTS: Forty-six patients received cefuroxime axetil (group C) and 44, phenoxymethyl penicillin (group P). The two groups differed in terms of age (patients in group C were younger), but no other differences in demographic and clinical pre-treatment characteristics were present. The clinical course during the post-treatment period revealed no significant differences between the two groups: the duration of erythema migrans (7.1 +/- 7.5 days in group C, 10.6 +/- 19.3 days in group P) and the appearance of minor manifestations of Lyme borreliosis (8.8% in group C, 9.1% in group P) were comparable; no major manifestations were recorded. Twelve months after antibiotic treatment all patients were free of symptoms. The patients treated with cefuroxime axetil had more drug-related adverse effects than did those treated with phenoxymethyl penicillin (26.1% versus 6.8%, p = 0.0301). "Herxheimer's reaction" at the beginning of treatment was identified more often in group C than in group P, but the difference was not statistically significant. CONCLUSIONS: Cefuroxime axetil and phenoxymethyl penicillin are equally effective in the treatment of children with solitary erythema migrans; however drug-related adverse effects were more frequently observed with cefuroxime axetil.