Comparative pharmacokinetics of metronidazole in healthy and Trichomonas gallinae infected pigeons (Columba livia, var. domestica).

1. This study investigated the pharmacokinetics of metronidazole after intravenous (i.v.) and oral administration to healthy and experimentally Trichomonas gallinae-infected pigeons, and determined the in vitro antiprotozoal activity of metronidazole against T. gallinae.2. Twelve pigeons which were experimentally infected to T. gallinae and twelve healthy pigeons received metronidazole at the dose of 25 mg/kg by oral or i.v. administration. Serial blood sampling was used for pharmacokinetic analysis. The metronidazole minimum lethal concentration (MLC) and the concentration killing 50% of the trophozoites (LC50) in the culture media were determined.3. In vitro data showed that the 24 h LC50 and MLC of metronidazole were 0.31 and 25 µg/ml, respectively. In vivo results showed no statistical differences between pharmacokinetics in infected and non-infected pigeons for both routes of administration. The area under the curve was statistically higher after the i.v. administration in both infected and healthy pigeons. The mean oral bioavailability was similar in the infected (83.8%) and the healthy (81.5%) birds.4. In conclusion, the pharmacokinetics of metronidazole in pigeons was not affected by experimentally-induced trichomoniasis. Despite in vitro susceptibility testing, which showed probable resistance of the isolated T. gallinae to metronidazole, five-day oral treatment of infected pigeons with 25 mg/kg metronidazole twice a day resulted in total eradication of trophozoites recovered in crop lavage of infected birds.

In vitro and in vivo activity of Artemisia sieberi against Trichomonas gallinae.

In Iranian folk medicine Artemisia sieberi has been used for treatment of parasite infections in human and animals. The present study was designed to evaluate the in vitro and in vivo effects of A. sieberi essential oil (EO) against Trichomonas gallinae. Trichomonas gallinae were recovered by wet mount method from infected native pigeons. The in vitro assays were accomplished in multi-well plates containing metronidazole (MTZ) as a standard antitrichomonal and EO in final concentrations of 2.5, 5, 10, 20, 50, and 100 µg/ml of culture medium containing 104 parasites. The invivo assay was performed on 40 experimentally infected pigeons receiving 25 and 50 mg/kg of MTZ and EO for 7 successive days. Gas chromatographic (GC) analysis was performed to reveal chemical constituents of the EO. At 20 µg/ml, MTZ resulted in no viable trophozoite in culture medium after 24 h incubation period. While the 24 h MIC of EO was 10 µg/ml. Treatment with EO at dose of 50 mg/kg after 4 days led to full recovery of infected pigeons but for MTZ at the same dose 5 days were spent. Major constituents of EO were α-thujone (31.5%) and ß-thujone (11.92%). Data of the present study introduced A. sieberi as a natural potent antitrichomonal agent effective against T. gallinae.

Antitrichomonal activity of Peganum harmala alkaloid extract against trichomoniasis in pigeon (Columba livia domestica).

1. This study was designed to evaluate the antitrichomonal effects of P. harmala alkaloid extract against T. gallinae, both in vitro and in vivo, as well as comparing it to that of metronidazole, conventional antitrichomonal medication and harmine and harmaline, the two alkaloids present in P. harmala. 2. T. gallinae were collected by the wet mount method from infected free-living pigeons. The in vitro assay was performed using multi-well plates containing test compounds in final concentrations of 5, 10, 15, 20, 30, 50 or 100 µg/ml. The in vivo assay was done on 60 experimentally infected pigeons dosed with metronidazole at 50 mg/kg body weight (BW) or alkaloids at 25 mg/kg BW. 3. The 24 h minimum inhibitory concentration (MIC) of alkaloid extract was 15 µg/ml while that of metronidazole was 50 µg/ml. Harmine and harmaline revealed 24 h MIC of 30 and 100 µg/ml, respectively. Treatment of infected pigeons with alkaloids led to a full recovery after 3 d but with metronidazole total eradication of trophozoites was not achieved. 4. In conclusion, data of the present study suggested P. harmala is a potent natural anti-trichomonal agent, effective against T. gallinae.

Molecular identification of macroscopic cysts of Sarcocystis in sheep in Babol, in the north of Iran.

Sarcocystis is one of the most prevalent parasite in domestic animals in the world. In this study, we examined 50 macroscopic cysts in sheep muscles from Babol, in the north of Iran. Genomic DNA extraction and polymerase chain reaction (PCR) were performed to amplify a 609bp length based on 18S rRNA gene. The results of restriction of AvaI, Hind II, TaqI and EcoRI enzymes demonstrated that all the samples were Sarcocystis gigantea. The results of this study supports the importance of molecular techniques for characterization of Sarcocystis species when valid preventive programs for identification and source of infection and progression of immunological diagnosis strategies are needed.

Serum total antioxidant capacity status of HTLV-1 infected patients.

Many aspects of the pathogenesis of Human T-cell lymphotropic virus type 1 (HTLV-1) still need further elucidations. Previous studies have indicated that oxidative stress occurs during infection with the other retrovirus, human immunodeficiency virus 1 (HIV-1). Similar results have been observed in some other chronic viral infections including hepatitis B (HBV) and hepatitis C (HCV). In order to reveal possible oxidative stress in HTLV-1-infected patients, we evaluated serum total antioxidant capacity (TAC) as an indicator of oxidative stress in these patients. Forty-four HTLV-1-seropositive individuals were included in this study, consisting of 12 symptomatic and 32 asymptomatic (carrier) cases. Controls consisted of 36 apparently healthy, HTLV-1-, HIV- and hepatitis-seronegative individuals. All symptomatic patients had HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Serum TAC levels in patients and healthy individuals were measured using a quantitative TAC assay. The antioxidant capacity in HTLV-1-seropositive cases was significantly reduced compared to control group (P = 0.001). In addition, TAC was lower in patients with more than 5 years history of HAM/TSP compared to those with ≤5 years duration of the myelopathy (P = 0.03). Our results show a depletion of TAC during HTLV-1 infection, which intensifies along with the disease progress. This finding indicates a role of the oxidative stress in pathogenesis of HTLV-1. These results may prompt further research to evaluate any possible therapeutic effect of antioxidant dietary supplements for HTLV-1 infected individuals.

Determination of the electrophoretic pattern of somatic and excretory-secretory proteins of Ligula intestinalis parasite in spirlin (Alburnoides bipunctatus).

Ligula intestinalis parasite is a cestodes that causes remarkable damages to fish. It is also of prime importance in economic and hygienic aspects. SDS-PAGE and western blotting are the methods that can be used to determine the electerophoretic pattern of somatic and excretory-secretory proteins of parasites. In this study, after obtaining the plerocercoidal stage of this parasite from the spirlin (Alburnoides bipunctatus), its somatic proteins were prepared using ultrasonicae, and excretory-secretory proteins were prepared using the PBS solution. After protein assay, which included using the Bradford method and then SDS-PAGE on these two antigenic solutions, 5 protein bands of 26, 33, 38, 58, 70kDa in somatic antigens, and 7 bands of 25, 28, 33, 43, 49, 60, 70kDa in excretory-secretory antigens were observed. After western blotting on both antigens and adding the primary antibody (the sera of infected fish) and then the secondary antibody (Rabbit Anti-fish Polyclonal Antibody Conjugated from Abnova Corporation) no band was seen in excretory-secretory antigen. And only in the 55kDa band of somatic antigen, a positive response, in comparison of fish positive serum was observed.

Determination of the electrophoretic pattern of somatic and excretory-secretory proteins of Ligula intestinalis parasite in spirlin (Alburnoides bipunctatus)

Ligula intestinalis parasite is a cestodes that causes remarkable damages to fish. It is also of prime importance in economic and hygienic aspects. SDS-PAGE and western blotting are the methods that can be used to determine the electerophoretic pattern of somatic and excretory-secretory proteins of parasites. In this study, after obtaining the plerocercoidal stage of this parasite from the spirlin (Alburnoides bipunctatus), its somatic proteins were prepared using ultrasonicae, and excretory-secretory proteins were prepared using the PBS solution. After protein assay, which included using the Bradford method and then SDS-PAGE on these two antigenic solutions, 5 protein bands of 26, 33, 38, 58, 70kDa in somatic antigens, and 7 bands of 25, 28, 33, 43, 49, 60, 70kDa in excretory-secretory antigens were observed. After western blotting on both antigens and adding the primary antibody (the sera of infected fish) and then the secondary antibody (Rabbit Anti-fish Polyclonal Antibody Conjugated from Abnova Corporation) no band was seen in excretory-secretory antigen. And only in the 55kDa band of somatic antigen, a positive response, in comparison of fish positive serum was observed.

Serologic evaluation of toxoplasmosis in matrimonial women in Babol, Iran.

To investigate anti-toxoplasmosis in matrimonial women, samples collected from who referred to Babol health center, that is the only pre-marriage laboratory guide center in Babol. This descriptive-cross sectional study has carried out on 241 matrimonial women to consider anti toxoplasmosis antibodies (IgG, IgM) by ELISA (Enzyme Linked Immunosorbant Assay). In this study, 241 women between 13 to 40 years old were considered. The average age was 20.98 years. Of these cases, 47.3% were urban and 52.7% were rural. 63.9% had IgG and 12.4% had IgM anti-toxoplasmosis antibody. This study carried out to determine anti toxoplasmosis antibodies in Babol in the first half of 2004. Dispersed studies have been done to determine the prevalence oftoxoplasmosis in Iran and mostly have reported a high and different prevalence oftoxoplasmosis in different parts of country.

Seroepidemiology of sheep toxoplasmosis in Babol northern Iran 2004.

This study is carried-out with the aim of determining the prevalence rate of toxoplasmosis in sheep at Babol (Bandpei part). In this study, 285 sheep of Bandpei area in Babol was selected. The samples studied with indirect immunofluorescence antibody. The results showed that 31.2% of sheep in this study were affected by toxoplasmosis. In positive cases, there was not any statistically significant difference between male and female sheep. The most positive cases observed by 1:200 titer. According to the researches in other parts of this country, the prevalence rate in this area is more than the others.

Differences in adsorption of serum proteins and production of IL-1ra by human monocytes incubated in different tissue culture microtiter plates.

In vitro cell culture models can be of great value in order to further analyze the regulatory mechanisms underlying the inappropriate function of the immune system in diseases such as autoimmunity and cancer. Cell culture conditions have to be well controlled in a way that they mirror the in vivo situation. The objective of this study was to compare tissue culture microtiter plates from different manufacturers with respect to their ability to support monokine production by human monocytes cultured in human serum. Tissue culture ware, made of polystyrene, undergoes treatment by the manufacturers to make the surface more suitable for culture of adherent cell populations. It is possible that quality differences in this treatment can lead to variations in protein binding properties and thereby influence the adherence and functional properties of monocytes. We measured spontaneous interleukin-1 receptor antagonist (IL-1ra) production by peripheral blood monocytes, cultured in human serum, in five different microtiter plates made for adherent cell culture. Culture in plates from two of the five manufacturers resulted in significantly lower amounts of secreted IL-1ra. IL-1ra release by human monocytes can be induced by adherent IgG cross-linking membrane receptors for the Fc part of IgG (FcgammaR). We found that reduced IL-1ra production coincided with a reduced capacity for binding of serum IgG in one case. Furthermore, this brand of microtiter plate also displayed the lowest level of adsorption of human albumin. We conclude that the protein adsorption properties of the plastic tissue culture ware have to be taken into consideration when assessing monokine production by human monocytes in vitro.

[Chronic fatigue syndrome: psychiatric perspectives]. / Le syndrome de fatigue chronique: aspects psychiatriques.

Chronic fatigue syndrome (CFS) is a common illness of unknown etiology and pathogenesis characterized by severe disabling fatigue and a variety of musculoskeletal, neurocognitive, mood symptoms and sleep disorders lasting at least six months. A significant proportion of patients fulfilling operative criteria for a diagnosis of CFS will also meet criteria for a psychiatric disorder such as depression, somatization or anxiety disorders. Premorbid lifestyle may play a predisposing, and/or perpetuating role in CFS. Some patients improve with time but most remain functionally impaired for several years. A variety of interventions have been used in the treatment and management of CFS. Interventions which have shown promising results include cognitive behavioural therapy and graded exercise therapy. Antidepressants can be useful particularly in the case of comorbid affective disorders. Development of good therapeutic doctor-patient alliance with empathic care is central to the effective management of CFS. In this article we overview the nature and definition of CFS. The prevalence, the prognosis and the psychopathological issues are presented. The management of this controversial syndrome is discussed.

Clinical impact of second harmonic imaging and left heart contrast in echocardiographic stress testing.

Second harmonic imaging and left heart contrast agents are recent echocardiographic advancements that enhance the assessment of wall motion. Because little information exists concerning their clinical impact on echocardiographic stress testing in daily practice, this was determined for 9-month periods before (1997) and after (1998) their introduction. Harmonic imaging was used in all patients after its introduction. A second generation intravenous left heart contrast agent (Optison) was used at the discretion of the sonographer and physician team. Both exercise and dobutamine stress tests were included. At the time of study interpretation, diagnostic confidence was assigned as high, medium, or low. For all patients who underwent coronary angiography < or = 6 months after stress testing, the diagnostic accuracy was determined (true positive plus true negative/total studies). There were 574 studies before and 746 studies after implementation. Optison was used in 28% of the harmonic imaging studies. Study cancellations due to uninterpretable images fell from 6.4% to 1.2% (p <0.001) despite a more obese population completing testing (body mass index: 29 +/- 7 to 31 +/- 8 kg/m2, p = 0.02), whereas high diagnostic confidence increased from 55% to 64% (p <0.001). For the 7% of patients who underwent cardiac catheterization, the diagnostic accuracy remained unchanged (74 vs 73%) although a prior negative stress test was less common (40% to 20% p = 0.04). Thus, these new technologies had a favorable clinical impact.

Factors leading to progression of valvular aortic stenosis.

The rate of progression of aortic stenosis (AS) in adults is variable. To determine whether clinical or echocardiographic variables are associated with more rapid hemodynamic progression, we identified 91 AS patients (initial valve area < or = 2.0 cm2) with 2 technically adequate studies separated by > or = 6 months. From the first study, left ventricular dimensions and AS severity were measured by standard Doppler-echocardiographic methods. Each aortic valve was graded for severity of calcification and degree of restricted leaflet motion; the sum of these grades provided a severity index reflecting leaflet pathology. Clinical and electrocardiographic variables were abstracted from medical records. Mean age was 68 years (range 29 to 89) and 61 were women. Initial AS severity ranged from an aortic valve area of 0.6 to 2.0 cm2 (median 1.3 cm2). During a mean follow-up of 1.8 years the aortic valve area decreased 0.04 cm2/year. The patient group with more rapid progression (decrease in aortic valve area > or = 0.1 cm2/year) had a larger proportion of men (p <0.01) and patients with an elevated serum creatinine (p = 0.04), a higher left ventricular mass index (p = 0.01), and a higher severity index (p <0.001). Multivariable regression analysis identified the severity index (direct relation) and the initial aortic valve area (inverse relation) as the only independent variables associated with more rapid progression. In conclusion, the rate of AS progression, although highly variable, is more rapid when leaflet calcification is more marked.

Embolic risk based on aortic atherosclerotic morphologic features and aortic spontaneous echocardiographic contrast.

BACKGROUND: Different aortic atherosclerotic plaque morphologic features may have varying embolic potentials. Spontaneous echocardiographic contrast (SEC) in the aorta, as in the left atrium, has been associated with an increased risk of embolic events and often occurs with complex aortic atherosclerosis. Thus an evaluation of their isolated and combined association with embolic events was undertaken. METHODS: We retrospectively studied all patients who underwent biplane or multiplane transesophageal echocardiography meeting the following inclusion and exclusion criteria: age >/=55 years and no other cardiac pathologic condition known to be associated with embolic events other than aortic atherosclerosis or aortic SEC. The 105 patients meeting the criteria were divided into those with aortic atherosclerosis and/or aortic SEC (the study group) and those without these aortic pathologic conditions (the comparison group). Complex aortic atherosclerosis was defined as mobile, ulcerated, or protuberant (> 4 mm). SEC was defined as a pulsatile, swirling echo pattern within the aortic lumen. Embolic events included strokes, transient ischemic attacks, or peripheral emboli. RESULTS: The 61 study patients and 44 comparison patients did not significantly differ with respect to the reason for referral, age, or sex. Embolic events occurred in 35 patients. Those with ulcerated or mobile plaques had a greater prevalence of embolic events (odds ratio 4.50; 95% confidence interval, 1.30-15.5; P <.05). The highest embolic event rate was seen in patients with any complex atherosclerosis and concomitant SEC (odds ratio 9.00; 95% confidence interval, 2.06-39.3; P <.01). Patients with SEC alone or protuberant plaques alone did not have a higher event rate (odds ratio 1.71 and 0.60; 95% confidence interval, 0.57-5.17 and 0.15-2. 47, respectively). CONCLUSIONS: Embolic events were associated with the presence of ulcerated or mobile aortic plaques. In addition, the combination of aortic SEC and any complex atherosclerosis had the highest embolic association.

Aortic sinotubular junction calcium as a marker of severe aortic atherosclerosis.

Because the significance of calcification of the sinotubular junction is poorly understood, we retrospectively investigated its association with aortic atherosclerosis in 101 patients who underwent transesophageal echocardiography. Such calcification was found to be a marker of more severe aortic atherosclerosis and, specifically, aortic arch disease.

Role of hydrazine in the mechanism of isoniazid hepatotoxicity in rabbits.

Isoniazid (INH) continues to be a highly effective drug in the chemoprophylaxis and treatment of tuberculosis; however, its use is associated with hepatotoxicity (predominantly hepatic necrosis) in 1-2% of individuals. The INH metabolites, acetylhydrazine and hydrazine, have each been implicated as the causative hepatotoxin in INH-induced hepatotoxicity. Using a model of INH-induced hepatotoxicity in rabbits, in which INH-induced hepatotoxicity manifests as hepatic necrosis, hepatic steatosis (hepatic fat accumulation) and hypertriglyceridaemia (elevated plasma triglycerides), we compared the severity of these measures of toxicity with plasma levels of INH, acetylhydrazine and hydrazine. Plasma INH and acetylhydrazine were not correlated with markers of INH-induced hepatic necrosis or fatty changes. Plasma hydrazine at 32 h was correlated significantly with plasma argininosuccinic acid lyase (ASAL, a sensitive marker of hepatic necrosis) activity as area under the curve (r2 = 0.54, P < 0.002) and log plasma ASAL activity at 48 h after the first dose of INH (r2 = 0.53, p < 0.005), but not with fatty changes. These results show in this model of INH-induced hepatotoxicity in rabbits that hydrazine, and not INH or acetylhydrazine, is most likely involved in the pathogenic mechanism of hepatic necrosis.

Mass-analyzed ion kinetic energy (MIKE) spectrometry and the direct analysis of coca.

The technique of mass-analyzed ion kinetic energy (MIKE) spectrometry is briefly reviewed. The direct analysis, without need of prior extraction or chromatographic purification, of cocaine and cinnamoylcocaine in coca plant tissues has been accomplished with this new technique. Detection limits of below 1 ng, with a quantitative accuracy of +/- 30% at the nanogram level, have been demonstrated.

Rapid determination of light halogenated hydrocarbons in urine.

We describe a rapid method for determining light halogenated hydrocarbons (LHH) in urine by electron capture gas-liquid chromatography. The hydrocarbons are extracted from urine into pentane. A 2-microliter injection of the pentane extract provides a detection limit of less than 1 microgram/liter.