Biological and biochemical studies for characterization of some Egyptian Trichomonas vaginalis isolates

Trichomoniasis has important medical, social and economical implications regarding its serious potentially associated complications, with the possibility of HIV acquisition and transmission. T. vaginalis is a very complex organism. Studying the variation in some biological and biochemical properties of the parasite can be used for characterization of the parasites. For the characterization of T. vaginalis infecting Egyptian female patients, the growth kinetics of 20 isolates, their pathogenicity, metronidazole susceptibility and electrophoretic protein patterns, were correlated with the recorded clinical manifestations associated with these isolates. Positive samples for T. vaginalis were cultured on modified Diamond`s medium. For growth pattern study, trophozoites were counted for each isolate every 24 hours for seven days. The pathogenicity assay was performed using intra-peritoneal inoculation in mice. The isolates susceptibility to different concentrations of metronidazole was recorded by determining the minimal lethal concentration [MLC] and trophozoites viability. The biochemical variability of the studied isolates was performed using 2-dimensional electrophoresis. A broad experimental variability was recorded among the 20 T. vaginalis isolates. There was a clear relationship between 3 isolates obtained from patients with severe vaginitis and the different parameters studied. These isolates had the highest number [20-25 organisms/HPF] in wet mount of vaginal discharge, and the shortest generation time [6:34-7:31 hours]; they were also highly pathogenic to mice. Only one isolate [no. 5] proved to be metronidazole resistant. The use of the first dimensional native-polyacrylamide gel electrophoresis [Native-PAGE] demonstrated the presence of some differences. The isolates were classified into two groups according to their proteins net charge. All samples in each group were considered as one isolate. However, when the 2-dimensional electrophoresis [SDS-PAGE] was applied, five different groups could be identified according to proteins molecular weights. There is a broad experimental variability among the studied Egyptian T. vaginalis isolates regarding growth kinetics, metronidazole drug susceptibility, degree of pathogenicity, as well as the electrophoretic protein patterns

Disinfection efficacy of sodium dichloroisocyanurate [NaDCC] against common food-borne intestinal protozoa

The present study was conducted to investigate the efficacy of sodium dich-loroisocyanurate [NaDCC] on the infective stages of common food-borne intestinal protozoa; Entamoeba histolytica [E. histolytica], Giardia lamblia [G. lamblia], Cryptosporidium, Cyclospora and Microsporidia; beside its effect on raw green vegetables and fruits. Parasites, isolated from stool of patients with diarrhea or dysentery, were exposed to NaDCC solution [1g/l] for one and two hours. Disinfection effect of NaDCC was assessed by in-vitro viability, using trypan blue stain, and infectivity bioassay in laboratory animals as indicated by fecal and intestinal parasitic counts. Raw vegetables and fruits were dipped in NaDCC solution in the same concentration and exposure time as used for treatment of the parasites. Results revealed statistically significant reductions in viability and infectivity of all examined parasites indicating their susceptibility to NaDCC. Relative variations in susceptibility were revealed; E. histolytica and G. lamblia were most susceptible [100% reduction] followed by Microsporidia then Cryptospridium and Cyclospora. NaDCC did not affect the consistency, color, taste or flavor of raw green vegetables and fruits. The proved efficacy of NaDCC, in cheap and convenient dry tablet form, makes it a promising tool in decontaminating raw vegetables and fruits from food-borne protozoan parasites at household and restaurant levels as well as in catering and fresh produce industry. It is also recommended for disinfection of food preparation surfaces and equipment

Immunization against trichinellosis using microwaved larvae of trichinela spiralis

The present study evaluated the effect of microwave irradiation on infective larvae of Trichinella spiralis [T. spiralis] by the ultrastructure changes of the microwaved larvae [ML] using scanning electron microscope [SEM]. The ML tested the ability to immunize mice against a challenge infection with T. spiralis. For the optimal dose and the best route of immunization inducing protection against challenge infection, two doses were used; 300 and 600 ML as one or two-dose regimen, each dose was given orally and intraperitoneally [IP]. SEM revealed tegumenttal damage of the ML in the form of distortion, loss of normal fold pattern and depressions or papillae protruded from their outer surface. After administration of the ML [orally or IP] to the non-infected control mice, neither adults nor larvae were detected in the intestines or muscles respectively. This indicated loss of larvae infectivity after exposure to the microwave irradiation. Also, a significant protection against challenge infection with T. spiralis was demonstrated in experimental mice immunized by ML, orally or IP. This was assessed by a statistically significant decrease in adult and muscle larval count, compared with the non-immunized infected control. Complete protection against both adults and larvae [100%] was achieved by IP injection of two doses of 600 ML, two weeks apart. The results suggested the feasible application of the microwave irradiation on meat for its decontamination from T. spiralis larvae. Such a method might be a promising a prophylaxis vaccine against trichinellosis in animals and/or humans

efficacy of antihelminthic compound; clorsulon against experimental schistosoma mansoni infection

The efficacy of Clorsulon[R] [CLS] against experimental schis-tosomiasis mansoni, using Praziquantel[R] [PZQ] as a therapeutic control was evaluated. Swiss Albino mice were divided into infected non-treated control, PZQ-treated group given a single dose of 500 mg/kg four weeks post infection [PI], and infected mice treated with single, double, and triple doses of 5 mg/kg CLS per dose, one week apart starting from the 4[th] week PI. All animals were perfused for adults count. Parts of livers and intestines were examined for granulomata number and sizes. Pathological changes in hepatic parenchyma by H and E and Masson trichrome stains were also examined. Results revealed that a single treatment with PZQ caused a significant percentage reducetion [%R] of worm load [92.68%], mean egg count in liver and intestine [91.20 and 94.01% respectively], and mean size of liver granulomata was reduced [92.06%]. Regarding CLS, the worm burden was reduced proportionally with number of doses given; 87.80, 96.34 and 97.56% in single, double and triple exposures successively. Egg count in liver was decreased by 85.90, 97.01 and 96.23% respectively in treated mice. Number of intestinal granulomata was decreased by 85.28, 94.24 and 95.49% in a similar way. Size of hepatic granulomata was decreased by 89.02, 94.51 and 95.05% by 1, 2 and 3 doses consecutively. All parameters reflected non significant difference between 2 and 3 dose of CLS. The results were critically discussed

Cardiovascular involvement in Kawasaki disease.

We report 72 patients with Kawasaki disease seen at this Centre over 7 years. Cardiac involvement in the form of mild pancarditis was seen in 28 % patients, but disappeared subsequently. Thirteen (18.5 percent) children developed coronary artery disease, out of which 4 resolved by the end of two months and another 6 after one year; 3 patients continued to show coronary artery dilatation and aneurysm formation. Children who received IV gammaglobulin in full dose within 10 days of onset of illness, showed no evidence of coronary artery disease during follow up.

Effect of acrylamide on the central nervous system of rates: histological, histochemical, and electron microscopy studies

The effect of acrylamide on the central nervous system of rats was studied in order to determine its neurotoxicity effect. Seventy rats were used in the present study which were divided into three groups. The rats of group one were used as control while rats of group two were given acrylamide by intramuscular injection with a dose of 5 mg/kg body weight, and the rats of group three were given 25 mg/kg acrylamide twice weekly for one month. The rats were sacrificed after 1, 2, 3, and 4 weeks of acrylamide injection and cervical spinal cord was removed and processed for histological, histochemical, and electron microscopical studies. Histological results showed mild degenerative changes of nerve fibers after one and two weeks of acrylamide injection while after three and four weeks severe changes were seen following both doses of acrylamide. Histochemical changes appeared in the decreased activities of cytochrome oxidase, RNA, and Nissl substances after one and two weeks of acrylamide injection and this decrease reached its minimal activity at the end of the experiment for the two doses while acetyl-cholinesterase and DNA were found to be increased. Electron microscopic study revealed irregular outline of nuclear envelop with some nuclear pores, vascular degeneration of cytoplasmic organelles, demyleinated and thinly myleinated axons, and many hypertrophied mitochondria. All these results indicated that acrylamide which is still used in the synthesis of polymers for a variety of industrial applications, is highly neurotoxic

Effect of radiotherpay and chemotherapy on normal jejunal mucosa of Swiss mice: histological, cytological, and histochemical studies

Radiation and chemotherapy play a major role in the management of patients with malignant tumours, they have marked toxicity against tumour cells in addition to other undesirable side effects on normal organs. This led the researchers to carry out a histological, cytological, and histochemical studies as regards the effect of radiation and chemotherapy on the jejunal mucosa of normal mice. Seventy-two male Swiss mice were used in this study. Golgi apparatus, acid phosphatase [ACP], and succinic dehydrogenase [SDH] enzymes were studied in normal jejunum as well as after exposure to radiotherapy and chemotherapy. The results revealed that prominent changes were observed in cisplatin-treated group, while more effects were shown in animals receiving combined radio and chemotherapy

Effect of acrylamide on albino rat kidney histological, histochemical and electron microscopy studies

The effect of prolonged intake of acrylamide on the kidney of the albino rat was studied. Forty adult male albino rats were used in this study to evaluate the possible structural toxic effect of chronic intake of acrylamide. This substance was administered orally in three doses [0.01, 0.05 and 0.5 mg/kg b.w] three days weekly for six months. Examination of paraffin sections using the light microscope showed variable histological changes in the different study gps. The changes detected proceeded badly with increasing the dose. Some of the renal tubules lost their cytoplasmic brush borders as well as acidophilic granules. Widening of Bowman's space was noticed. Also, the interstitium and glomeruli showed lymphocytic infiltration and more congestions. Histochemical study revealed decreased glycogen and increased DNA contents. Electron microscopy studies showed aggressive changes in the nuclei, mitochondria and microvilli of kidney of rats [0.5 mg/kg]. These results indicated that chronic intake of acrylamide has a toxic effect on the kidney tissue cells when given with 0.5 mg/kg for six months as indicated by histological, histochemical and electron microscopic changes

utility of lactoferrin in differentiating parasitic from bacterial infections

Lactoferrin is an iron binding glycoprotein found in the 2 ry granules of PMN. In order to determine the usefulness of such marker for neutrophilic activity in differentiating cases suffering from amoebic and bacillary dysentery, Schistosoma and bacterial UTI infections, we examined stool and urine specimens using anti-lactoferrin antibodies [lactoferrin latex agglutination test: LFLA], compared with different standard gold techniques. Our results demonstrated that cases with either shigllosis or UTI revealed a high lactoferrin titer which was positively correlated with the number of PMN. In addition cases with Entamoeba histolytica or S. haematobium were characterized by relatively lower inflammatory process as expressed by mild lactoferrin titer which was also correlated with the PMN count. In addition, the findings of the present work indicated that LFLA was sensitive and specific when used alone and its sensitivity was augmented after coupling with other simple indirect methods of diagnosis. In conclusion, results described the reliability of using LFLA as a simple, rapid, sensitive method in differentiating, certain parasitic from bacterial diseases

Significance of circulating intercellular adhesion molecules [sICAM-1] and thrombopoietin [TPO] in hepatitis C vinis related liver cirrhosis

Chronic hepatitis C is a major health problem that causes mortality in the worldwide distribution. The systemic levels of many soluble adhesion molecules and cytokines are altered in autoimmune diseases and liver infection. Among mediators that show altered serum levels are sICAM-1 expressed by immunocompetent cells and thrombopoitin [TPO]. The aim of this study was to evaluate the serum sICAM and TPO and their correlations with clinical and laboratory data in patients with hepatitis C-related cirrhosis. 40 patients with HCV-related cirrhosis and 20 healthy subjects were enrolled in this study. They were subjected to the following investigations: complete blood picture, liver and kidney functions, prothrombin time[PT], abdominal sonography and estimation of serum TPO and sICAM-1. Compared to controls, the patients group showed significant increase in spleen size [P<0.025], portal vein diameter [P<0.0025], ALT, AST, bilirubin, sICAM-1 [P<0.0005], and PT [P<0.05]. However there was significant decrease in serum albumin levels [P<0.01], platelets count [P<0.0005], RBCs count, haemoglobin concentrations [P<0.025] and serum TPO levels [P<0.0025]. The serum creatinine levels and WBCS count showed insignificant changes with control. The serum TPO levels were negatively correlated to ALT [r=-0.97, P<0.0001], PT [r=-0.96, P<0.0001], spleen size [r=-0.95, P<0.0001], and portal vein diameter [r=-0.96, P<0.0001] and positively correlated to platelets [r=0.84, P<0.0005]. While serum sICAM-l were positively correlated to ALT [r=0.99, P<0.0001], PT [r=0.94, P<0.0005], spleen size [r=0.95, P<0.0001], and portal vein diameter [r=0.85, P<0.005] and negatively correlated to platelets count [r=-0.7l, P<0.005]. Our results suggested that impaired synthesis of TPO by cirrhotic liver may contribute to the development of thrombocytopenia and related to liver cirrhosis together with increased splenic sequestration of platelets by the enlarged spleen. So recombinant human TPO should be evaluated in the treatment of thrombocytopenia in HCV-related cirrhosis. In addition, sICAM-1 elevation in plasma of patients suffering from HCV-related cirrhosis was related to the degree of cirrhosis and portal hypertension. So, sICAM-1 may be used as a marker of the disease activity and may provide diagnostic and prognostic information. However this needs to be further studied to detect the cut off level of sICAM-1 in Egyptian HCV-related liver cirrhosis

Effect of sun exposure on the skin: histological, histochemical, and immunohistochemical studies

The present work was designed to explore certain aspects concerning human skin in the domain of sun exposure. Skin biopsies were taken from fifteen subjects divided into two groups; chronically sun-exposed group consisted of 10 subjects and control group consists of 5 subjects with no history of chronic sun exposure. Histological signs of sun-damaged skin were prominent as a pronounced thickening of stratum corneum indicating a hyperproliferative state accompanied by a corresponding decrease in collagen content associated with large bundles of coarse abnormal elastic fibers. Depletion of Langerhans immune cells in addition to decreased nuclear DNA were also observed

Electron microscopical studies of pituitary gland of male hamsters under the effect of estrogen treatment

The ultrastructure of the normal pars intermedia of male hamsters showed two types of cells. The light cells were numerous, regular in shape with rounded or elongated nuclei have different size of secretory granules and were considered as a source of melanocyte stimulating hormone [MSH]. The dark cells were less in number, showed less display of secretory granules and were concerned with colloid production. Some of the dark cells showed foot like processes and irregular nuclear membranes. Progressive changes in the cell organelles were observed in the pars intermedia cells after long term of estrogen treated hamsters over the studied period

Study on certain serum glycosylated proteins and lipids in diabetic retinopathy

The aim of the present work was to through more light on some factors involved in the vascular complications in diabetes mellitus; namely, plasma fibronectin and plasma alpha-2-macroglobulin in diabetics with normal fundus and in diabetics with retinopathy in a trial to clarify their possible role. Fifty diabetic patients [30 with retinopathy and 20 without retinopathy], in addition to 20 healthy control subjects were included in the present study. The study concluded that alpha-2-macroglobulin is a valuable indicator in the development of angiography in general than fibronectin, since the former was proved to be significantly elevated in diabetics with retinopathy than without, in addition to its significant correlation with microalbuminuria. Fibronectin showed neither elevation in retinopathic diabetics nor any correlation with microalbuminuria

Increased production of vascular endothelial growth factor and some other cytokines in serum and ascitic fluid of cirrhotic patients with spontaneous bacterial peritonitis

The aim of this study was to determine serum and ascetic fluid vascular endothelial growth factor [VEGF], tumor necrosis factor- alpha [TNF-alpha], interleukin-6 [IL-6] and endotoxin, in cirrhotic patients and to search for a relationship between them and SBP. This study was performed on 40 cirrhotic patients with SBP, their ages ranged from 35 to 60 years, with mean of 47.5 +/- 11.6 years, 40 cirrhotic patients with non-infected ascites their ages ranged from 22 to 50 years, with mean of 34.5 +/- 6.5 years and 20 healthy control subjects with matched age and sex. From the results obtained it was concluded that VEGF, IL-6, TNF-alpha and endotoxin in the serum and ascetic fluid may become useful markers for diagnosis of SBP and for monitoring the treatment of cirrhotic patients

rapid technique for diagnosis of intestinal parasitic infection

This study was carried out on stool specimens of 150 cases attending the outpatient clinics of the Faculty of Medicine, Alexandria University. All patients were suffering from gastro-intestinal disturbance specially diarrhoea. Each specimen was examined for intestinal parasites by direct saline and iodine smears and was stained by using: quantitative buffy coat [QBC] tube technique, modified trichrome and modified Ziehl Neelsen stains. This work aimed to evaluate the quantitative buffy coat tube method in diagnosis of parasitic infection in such cases, comparing it with the other staining reference techniques used simultaneously. Out of 150 diarrhaeic patients 67 were positive for protozoa. These protozoa identified in the examined specimens were Giardia lamblia, Cryptosporidium parvum, Cyclospora, Microsporidia and Blastocystis hominis. The results showed high sensitivity and specificity [100%] of the QBC tube method over the other staining techniques

Histopathological, ultra-structural and histochemical studies on the liver of schistosoma mansoni infected mice under the effect of triclabendazole

The present work investigated the effect of the fasciolicidal drug triclabendazole [TCBZ] on the liver of S. mansoni infected mice. The drug was given to normal as well as to S. mansoni infected mice. The work included histopathological, ultra-structural and histochemical studies. In the normal liver TCBZ induced the formation of scanty foci of inflammatory infiltrate. No changes were observed at the subcellular level or in the hepatocytes enzymes succinic dehydrogenase [SDH] and acid phosphatase [ACP]. The liver of S. mansoni infected mice revealed the classical histopathological picture of schistosomiasis. After TCBZ treatment, the granulomata involuted revealing fibrous transformation. The ultrastructural of hepatocytes of S. mansoni infected mice revealed distortion of the mitochondria, increased number of lysosomes and obliteration of Disse space. These electron microscopic [EM] changes were less obvious after TCBZ therapy denoting improvement of the hepatocellular insufficiency. Histochemically, an increase in ACP and a decrease in SDH activity were observed in S. mansoni infected liver. The activity of these enzymes returned to normal after treatment with TCBZ. It could be concluded that TCBZ has no direct toxic effect on the hepatocytes. In experimental schistosomiasis TCBZ improved the liver pathology and enzymatic activity of the hepatocytes

role of foliar fertilization on the growth and productivity of pea [pisum sativum L.] plants cultivated under different sowing methods

Two field experiments were carried out at the Experimental Station of the National Research Centre, during 1993 and 1994 seasons to study the response of pea [Pisum sativum L.] plants to the treatments of foliar fertilizers under different sowing methods. The obtained important results are as follows: Wide sowing of pea, i.e. 2 lines/ridge resulted the highest plant growth characters, i.e. the higher number of leaves, dry weight of shoots number of pods/plant and average pod weight. In spite of this the more early as well as the total pod yield were obtained with the dense sowing, i.e. 3 lines/ridge. Within the used mineral fertilizers, ammonium nitrate [33.5 percent] +galafertile as a mixture when sprayed 3 times induced the best growth and yielded the heaviest early and total pod yield as weft as improving the quality of pods. Also, the data showed that the growth, yield and its quality were depended more on the foliar fertilizer treatment than on the sowing methods