Ultrastructural alterations in Schistosoma mansoni juvenile and adult male worms after in vitro incubation with primaquine

BACKGROUND Praziquantel has been cited as the only drug for treating schistosomiasis. However, concerns over drug resistance have encouraged the search for novel drug leads. The antimalarial drug primaquine possesses interesting anti-schistosmal properties. OBJECTIVES This study is the first to document the potential role of primaquine as a schistosomicide and the ultrastructural changes induced by primaquine on juvenile or adult male worms of Schistosoma mansoni. METHODS Ultrastructural alterations in the tegumental surface of 21-day-old juvenile and adult male worms of S. mansoni were demonstrated following primaquine treatment at different concentrations (2, 5, 10, 15, and 20 µg/mL) and incubation periods (1, 3, 6, 24, and 48 h) in vitro, using both scanning and transmission electron microscopy. FINDINGS At low concentrations (2, 5, and 10 µg/mL) both juvenile and adult male worms were alive after 24 h of incubation, whereas contraction, paralysis, and death of all worms were observed after 24 h of drug exposure at 20 µg/mL. The tegument of juvenile and adult male worms treated with primaquine exhibited erosion, peeling, and sloughing. Furthermore, extensive damage of both tegumental and subtegumental layers included embedded spines, and shrinkage of muscles with vacuoles. The in vitro results confirmed that primaquine has dose-dependent effects with 20 µg/mL as the most effective concentration in a short incubation period. MAIN CONCLUSIONS The schistosomicidal activity of primaquine indicates that this drug possesses moderate in vitro activity against juvenile and adult male worms, since it caused high mortality and tegumental alterations. This study confirmed that the antimalarial drug primaquine possesses anti-schistosomal activity. Further investigation is needed to elucidate its mechanism of action.

Histopathological insight on malathion induced nephropathy which is ameliorated by antioxidants USE; an animal model

Background and aims: Malathion induced nephro-toxicity is a challenging process, several mechanisms were contributed. The present study was designed to evaluate the histopathological changes evoked by malathion in renal tissue compared to those induced by malathion plus antioxidants, to clarify the underlying mechanisms, using immunohistochemical staining of proliferating cell nuclear antigen [PCNA] and CD34

Methods: Forty adult male albino rats were divided into four equal groups. Group 1; control. Group 2 received malathion [27 mg/kg/day] orally. Group 3 received malathion plus vitamin E [400 mg] once/day, orally. Group 4 received malathion plus vitamin C [100 mg,] once/day, orally. Animals of all groups were sacrificed after 2 months. Histopathological assessment [H and E and Masson 's trichrome], PCNA and CD34 immunostaining were performed to all groups

Results: Group 2 showed a spectrum of histopathological changes; collapsing glomerulopathy with focal sclerosis, degenerative tubular changes and diffuse vascular congestion. Both groups 3 and 4 showed marked reduction of previous changes. Masson 's trichrome stained sections showed more collagen deposition in group 2 compared to groups 3 and 4. Group 2 revealed a decrease in the mean number of total PCNA positive cells [20. 8 +/- SD 5.7] [P< 0. 001] compared to control group. Groups 3 and 4 showed high significant increase in the mean number of total PCNA positive cells [31.3 +/- SD 5.7 and 31.2 +/- SD7.8] [P<0.001 and 0.004, respectively]. CD 34 immunoslaining of group 2 showed negative to minimal expression compared to moderate to strong intensity of groups 3 and 4

Conclusion: Malathion can induce a unique pattern of collapsing glomerulopathy with wide spectrum of histopathological changes. Podocytopathy is the main insult which began with podocytes proliferation and hypertrophy and ended by podocytopenia and glomerular sclerosis. The antioxidant use [vitamins E and C] provides a protective role against the previous nephrotoxic changes. PCNA imnmunostaining highlighted the role of injured podocytes while CD34 ascertained the structural and/or antigenic alteration of endothelial cells in collapsing glomnerulopathy

Optimization of polymerase chain reaction for direct detection of colibacillosis in infected chickens

A total of 33 local E. coli isolates were used in this study. These isolates were biochemically and serologically identified as O1, O2, O6, O78 and O126. Four sets of oligonucleotide primer sequences were designed specifically for 16SrRNA, STX, Sth and eaeA genes. DNA and Plasmids were extracted and polymerase chain reaction was optimized for each. 16SrRNA gene primer successfully amplified with all serotypes giving rise a product mass of 204 bp, while STX gene primer was amplified with O1, O2 an dO78 serotypes in a specific band at 323 bp. At the same time the specific primers of Sth gene get a 171 bp molecular weight product only with O6 serotype meanwhile the eaeA gene primers successfully amplified only with O126 serotype giving rise a molecular weight band at 200 bp. In conclusion, PCR assay was able to differentiate between the different serotypes of E. coli in the suspected samples saving time, money and effort

Effect of exogenous ascorbic acid on wheat tolerance to salinity stress conditions

In vitro experiments were performed to determine responses of wheat calli to ascorbic acid [AsA] concentrations [0, 250, 500, 1000 and 2000 ppm] under different levels of sea water [0, 15, 30 and 45%] and to determine suitable concentrations of AsA for exogenous treatments to enhance wheat tolerance to salinity. Results of this study indicated that AsA of [500, 1000 and 2000 ppm] concentrations improved tolerance of wheat calli to salinity. Two pot experiments were conducted during the two successive seasons 2004-2005 and 2005-2006 to determine the effect of exogenous AsA of 0, 500, 1000 and 2000 ppm concentrations at 45 and 75 days after sowing on growth, chemical composition and yield of wheat plants cv. Giza 168 [salt sensitive] irrigated with different levels of sea water [0, 15, 30 and 45%] in comparison with plants of cv. Sids 1 [partially salttolerant]. The obtained results in this study clearly proved the role of AsA in increasing tolerance of wheat plants to salt stress. Exogenous AsA enhanced plant growth, chemical composition and consequently the productivity of wheat plants under salinity stress conditions. These effects may be attributed to the protective role of AsA in plant cells from the oxidative stress induced by salinity. It could be concluded that exogenous AsA of 1000 ppm concentration at 45 and 75 days after sowing is the most effective treatment to increase wheat tolerance to salinity

relation between the duration of inhalation abuse of a volatile substance and its effect on the testis. a histological and biochemical study in adult male albino rats

Volatile substance abuse in general, and toluene inhalation in particular, for their neuropsychological effects, represents a significant problem in many developed and developing countries. The present work was designed to investigate the histopathological changes in the testis of adult male albino rats, induced by toluene vapour inhalation over different periods. The present study was carried out on forty adult male albino rats with body weights ranging from 60-100g. The animals were categorized into two groups: Group I: [Control Group] included ten rats received no treatment, Group II: [Toluene inhalants] included thirty adult rats exposed to toluene vapour inhalation. A clean dry piece of cotton was soaked with toluene liquid and placed in the covered cages three times daily, each for about thirty minutes for six days per week. These animals were subdivided into three equal subgroups according to the exposure period; Subgroup [A]: ten rats exposed to toluene vapour for two weeks, Subgroup [B]: ten rats exposed to toluene vapour for eight weeks, Subgroup [C]: ten rats exposed to toluene vapour for twelve weeks. At the end of each duration of the experiment, animals were scarificed by decapitation using light ether anesthesia after taking blood samples. I- Histological examination: Specimens were taken from the testis of all rats and processed for examination by light microscope using haematoxylin and eosin stain and ultrastructural study using the transmission electron microscope. II. Hormonal assay: The concentration of testosterone level, luteinizing hormone [LH] and follicle - stimulating hormone [FSH] were estimated by radio immunoassay. III. GAS chromatography: Concentration of toluene vapour in the blood was measured by High performance liquid chromatography. IV. Statistical analysis: The one way ANOVA test was applied to estimate the significant values of the hormonal assay for serum testosterone, LH and FSH and the 5% level of significance was chosen. The histopathological changes observed in the testis of rats exposed to toluene inhalation demonstrated its potentials to induce cytotoxic effects on the spermatogenic cells, Sertoli cells and the interstitial cells of Leydig. The severity of the toluene damaging potentials appeared to be dependent on and directly proportionate to the duration of toluene inhalation. So, the histological changes were mild and scattered in the testis specimens of group A [2 weeks inhalation] and was more severe in both eight and twelve weeks groups. The correlation between high performance liquid chromatography for toluene gas in blood, the biochemical gonadal and gonadotrophin hormonal assay and the histological assessment, explored the various mechanisms that were incorporated in the establishment of the toluene induced testicular injury. The present study proved the undoubting evidences for the damage potentials of toluene on the testis as the major reproductive organ in the male. Furthermore, the study showed the direct proportionality between the toxic effects of toluene vapor and the length of the exposure duration. Yet, the observed histological alterations were highly suggestive for a probable impaired reproduction in experimental animals which needs further study

Prevalence of glucose abnormalities in patients with hepatitis C virus infection

In addition to causing serious liver disease [cirrhosis and liver cancer], HCV infection can also contribute to the development of some diseases outside the liver among these, diabetes mellitus. [DM]. the frequency of type 2 diabetes mellitus tends to be higher in patients infected with hepatitis C virus [HCV] than those without HCV infection [Mehta et al., 2000]. The study was carried on 70 individual, 60 were HCV positive patients and 10 apparently healthy persons. We found that 31.6% of the patients were diabetic, 45% of the patients have impaired glucose tolerance, and 23.4% patients with normal blood glucose. There was a highly significant increase in both C-peptide and blood insulin levels in diabetic patients when compared to patients with impaired glucose tolerance and patients with normal blood glucose, p-value 0.0001

Results of senning operation modified with wide-based pericardial graft [an echocardiographic evaluation]

Despite the fact that the arterial switch operation is the treatment of choice for complete transposition of the great arteries, patients with unbalanced ventricles still needs the Senning operation. Known surgical complications like baffle obstruction may result in early deterioration of ventricular function and impaired exercise performance. It is not clear wether these complications are inevitable or predisposing factors at surgery or during follow up may identify patients at high risk Baffle obstruction after Senning operation may result in early deterioration of RV function and impaired exercise performance. The Objective was to assess the effect of surgical modification in avoiding surgical complications with its deleterious effects on early as well as late follow up. Echocardiography was used to assess postoperative outcome. This prospective study included 30 cases who were candidates for Senning operation for simple transposition of great arteries in the cardiothoracic department of Aboul-Reesh Pediatic hospital Cairo University where an in-situ wide-based pericardial patch was used for the outer baffle, modified right atriotomy and suture lines were modified in all patients. Left atrial appendage augmentation was used in 8, Eustachian valve in 6 cases. Postoperatively patients were subjected to clinical evaluation using Ross scoring for heart failure, ECG and echocardio graphic evaluation to assess RV size, degree of TR, baffle patency, LVOT dynamic obstruction, and global RV systolic and diastolic functions using the myocardial performance index [MPI]. We had early 2 mortalities [6.66%] out of low cardiac output, accelerated nodal rhythm was experienced in other 2 patients [6.66%], internal baffle pressure was 6.1 +/- 2.64cc H20. Data higher than 12cc H[2]O was found in 3 patients [10%], none of them had augmentation with the left appendage. Outer baffle pressure was 9.2+ 1.08 cc H20. No baffle leakage was detected. Clinical examination postoperatively revealed that only one patient had four points on Ross score of HF and the rest of patients had a zero score. Nodal arrhythmias were found in three patients, marked RV dilatation was found in one case and the other 27 cases had mild dilatation, moderate TR was present in 4% of cases, mild TR in 72% of cases. There was no baffle obstruction or LVOT obstruction in our patients. Patients who had Senning operation after the first year of life [40%] had RV MPI more than 0.4 while those under the first year [60%] had MPI less than 0.4. Baffle Augmentation with the wide based pericardial patch offered less chance for baffle obstruction, Left atrial appendage could be used in addition to offer an additional tool to avoid higher internal baffle pressures. Patients subjected to Senning operation earlier in the first year of life and patients have higher oxygen saturation at the time of the operation have better global RV functions

Myocardial performance in hydroxyurea versus transfusion treated sickle cell disease pediatric cases

Sickle cell disease [SCD] is associated with many cardiac abnormalities including increase cardiac output, pulmonary hypertension [PH] and myocardial perfusion abnormalities. Hydroxyurea [HU] is considered to be the most successful drug therapy for severe sickle cell disease. The objective is to study the echocardiographic cardiac abnormalities in patients with SCD on HU therapy comparing them with those on isolated blood transfusion regimen. Descriptive cross sectional case study was conducted on 29 cases of SCD on transfusion regimen [16 cases] and on HU therapy [13 cases]. Doppler echocardiographic assessment of left ventricular systolic and diastolic function, calculation of myocardial performance index [MPI] and left ventricular mass were done to all patients. Right ventricular MPI was higher in HU treated group with difference approaching significance [p = 0.06]. No significant correlation between HU dose and duration and left ventricular systolic or diastolic function, right ventricular diastolic function, or pulmonary artery pressure [PAP]. PH was present in 55% of SCD. No significant difference in PAP between HU and transfusion treated groups. Negative significant correlation between left ventricular deceleration time and estimated systolic pulmonary artery pressure [r=-0.37 p =0.04]. No deleterious echocardiographically detected cardiac effects were observed in HU treated SCD patients. HU had no effect on PH

Selenium and thyroid hormone status pre and post open heart surgery in children with congenital heart disease

Transient myocardial ischemia that occurs during cardiac surgery leads to oxidative stress and production of free radicals. Selenium [Se] status has important implications for cardiovascular morbidity and mortality as selenium is an integral component of the enzymes glutathione peroxidase and 5-iodothyronine deiodinase. Thyroid hormones have important effect on the heart and peripheral vascular system. Cardiopulmonary bypass [CPB] results in low triiodothyronine [T3] state, due to central depression of the hypothalamic pituitary axis together with impaired conversion of T4 to T3 secondary to reduced activity of iodothyronine deiodinase. This work aimed to study the effect of cardiopulmonary bypass on selenium and thyroid hormone status in infants and children with congenital heart disease [CHD]. Also it aimed to study the possible effect of selenium and thyroid hormone status on the hemodynamic condition of these infants and children after open heart surgery. Twenty five children were subjected to open heart surgery in the Cardio Thoracic Unit, Pediatric Hospital, Cairo University. The patients were subjected to full history taking, general and local cardiac examination, M mode, 2D and Doppler Echocardiography. Serum samples were collected, at the induction of anesthesia and 48 h after cardiopulmonary bypass for assessment of selenium, free T3[FT3]and free T4[FT4] levels. Assessment of serum selenium takes place at [NRC] using graphite furnace atomic absorption spectrometry [GFAAS]. Assessment of FT3 and FT4 takes place at Radioactive Isotope Department using RIA technique. Significantly lower postoperative selenium, FT3 and FT4 serum levels, compared to those at the induction of anesthesia, were found [p< 0.005, 0.025, and 0.025 respectively]. Longer CPB time was associated with lower FT3 and FT4 levels [r =0.49 and 0.63 respectively]. On the other hand higher FT3 and FT4 levels were associated with less time needed for inotropic support [r =0.48 and 0.60 respectively]. There is significant decrease of selenium, free T3 and freeT4 levels after cardiopulmonary bypass. The degree of decrease of thyroid hormones is affected by CPB time. Thyroid hormone levels have an evident impact on the myocardial functions and hemodynamic condition after open heart surgery

Doppler echocardiographic assessment of aortic stiffness and myocardial performance index in insulin dependent diabetics

Type I diabetes is characterized by diffuse arterial wall stiffening and thickening which progress with the severity of the disease but could be detected also in the absence of any diabetic-related complications. The stiffening of aorta and other central arteries is a potential risk factor for increased cardiovascular morbidity and mortality. In this study, we aimed to assess the aortic stiffness index, aortic strain, aortic pulse wave velocity [PWV] and myocardial performance index [MPI] in insulin dependent diabetic children and adolescents non invasively using Doppler echocardiography, correlating it with disease duration, state of glycemic control, and lipid profile. Thirty normotensive insulin dependent diabetics and 33 healthy control group were studied. 2D, M-mode and Doppler echocardiography was performed. Calculation of aortic strain, aortic stiffness index and MPI were done. Measurement of glycosylated hemoglobin [HbA1c] and microalbuminuria were performed. The results proved that the aortic stiffness index and aortic strain were significantly higher in diabetics than in the control group [p=0.02] and [p=0.001] respectively. Aortic strain correlated positively with HbA1c [p=0.05, r=0.4]. Aortic PWV and MPI were significantly higher in the cases [p=0.0001] and [p=0.003] respectively

Conclusion: Insulin dependent diabetic children and adolescents had stiffer aortic arteries than control subjects, Aortic strain correlated positively with state of glycemic control. Myocardial performance index was significantly higher in the cases in spite of the normal ejection fraction that may denote an early sign of left ventricular systolic dysfunction

Role of chest ultrasound in determining the nature of pleural effusion in children

Because of its ready availability and ability for bed side imaging, sonography has become a major imaging modality not only in determining the presence of pleural fluid and its nature, but also as a guide to aspiration. The pleura is a relatively superficial structure and therefore, it is accessible to sonography. The aim of our work was to investigate the usefulness of sonographic findings, including the internal echegenicity and associated changes in the pleura and lung parenchyma, in determining the nature of pleural effusion, and to compare the results with the CT findings, and those of pleural aspirate. This study was conducted on 50 children suffering from pleural effusion ranging in age between 2 months and 13 years. They were subjected to full history taking, clinical examination, laboratory investigations, laboratory analysis of pleural aspirate, chest Xray, CT chest [if needed] and chest ultrasound. The results of our study revealed that pleural fluid appearance in chest ultrasonography was classified into four main categories: anechoic in 50%; complex non septated in 16.7%; complex septated in 8.3%; and homogeneously echogenic in 25% of cases. We regarded transudates [45.8%] as usually anechoic. Exudates [28.2%] were considered when the pleural effusion was complex septated and non septated. Empyema [20.8%] was considered in our study in cases with homogeneously echogenic pleural effusion. Chest ultrasonography detected 91% of transudates, 80% of empyema, and all cases with inflammatory exudates

Conclusion : We concluded that chest ultrasonography is a valuable method in assessing the nature of pleural effusion and it seems to give additional information over chest x-ray and CT chest

Rapid flow cytometric susceptibility testing of candida species

The present study was performed to establish a standardized rapid antifungal susceptibility method using flow cytometry [FCM]. Also, to detect the antifungal susceptibility pattern of Candida species isolated in Ain Shams University Hospitals. For this purpose, 50 Candida isolates from different patients' specimens [urine, sputum, pus, blood and ascitic fluid] were identified to the species level using Microscan Walkaway 40. Their in vitro susceptibility to amphotericin B [AMB], fluconazole and ketoconazole was performed using 2 reference broth microdilution NCCLS M27-A and FCM. Membrane permeability changes of Candida caused by antifungal agents were measured by FCM using propidium iodide. The most common isolates were C. albicans [48%], C. Tropicalis [40%], and others: C. parapsilosis, C. Lusitaniae, C. glabarata and C. Krusei [12%]. By both methods [M27-A and FCM], Candida species were 90% and 92% susceptible to AMB respectively; 80% and 4% susceptible to ketoconazole respectively and 84% and 88% susceptible to fluconazole respectively. On comparing results of both methods for antifungal susceptibility testing, it was found that by FCM, a 4 hrs incubation for azoles and 2 hrs incubation for AMB were aufficient to provide MICs comparable to the reference M27-A broth microdilution method developed by NCCLS. Subcommittee on antifungal susceptibility tests [correlation significance < 0.001]. Such work showed that FCM provides rapid and sensitive method for antifungal susceptibility of candida

Autolytic changes in blood cells of albino rats a post-mortem morphologic study

The morphology of various types of cells in the peripheral blood of albinorats cadavers was investigated. The material comprised 32 adult male albinorats from which ante-mortem blood samples were compared to postmortem samplestaken every hour up to 6 hours after death. The rat corpses were eitherexposed to the environmental temperature [31 +/- 3C], or kept at +10C in anincubator. The red cell count, and the total white cell and platelet countsdecreased significantly during the first few hours postmortem in theenvironmentally-exposed group of corpses. Red cells were quite rapidlytransformed from the normal rounded discoid configuration to acquireinvaginations, crenations and later on, spheroidal configuration and membraneabsorption. Rapid deterioration of the staining properties, and markedmorphological changes in many leucocytes occurred quite rapidly after death. Lymphocytes seemed to be the most resistant to the effect of autolysis. Thepresent morphological observations and the quantitative results suggested thatvarious cellular elements of the blood seem to be quite resistant to autolyticeffects in the early postmortem period, and many cells apparently retain theirnormal morphology for longer periods of time in the blood of rat cadavers keptat reduced temperature