Efficacy of black seed oil from Nigella sativa against murine infection with cysts of Me49 strain of toxoplasma gondii

New therapies for toxoplasmosis are critically needed. Nigella sativa, commonly known as black seed or black cumin, has been known to include many medicinal properties. It has anti-inflammatory, and immuno-potentiating effects, antihelminthic and antiprotozoal activities. To study the effect of black seed oil [BSO] from Nigella sativa against Toxoplasma gondii Me49 strain in a murine model of infection. Two separate studies were performed, in which mice were orally inoculated with 10 or 20 T. gondii [Me49 strain] brain cysts. In each study, three groups of mice [35 each] were assigned to treatment with BSO for 2 weeks before T. gondii infection [BSO prophylactic], day 4 post infection [BSO therapeutic], or left untreated [infected untreated control]. The BSO effect on toxoplasmosis was evaluated by the assessment of [1] survival rate and brain cyst burden, [2] brain histopathological lesions and [3] immunohistochemical expression of inducible nitric oxide synthase [iNOS]. In infection induced by inoculation of 10, but not 20, cysts/mouse of the Me49 strain, BSO in prophylactic or therapeutic regimens significantly enhanced protection of infected mice against death [P = 0.01] and reduced brain cyst burdens at 5, 7 and 12 weeks post infection [PI] [P < 0.05] compared to the infected untreated control. The brains of BSO prophylactic or therapeutic groups showed milder meningitis, encephalitis and perivascular cuffing compared to the infected untreated control [P < 0.05]. Moreover, expression of iNOS was significantly enhanced in both BSO prophylactic and therapeutic groups compared to the untreated infected control. The BSO prophylactic group showed a significant enhanced expression of iNOS, selective to the brain endothelial cells, in the 1st week PI. Infection with 20 cysts was more aggressive, resulting in death of all untreated mice by day 35, and 26.7% and 20% protection respectively in BSO prophylactic and therapeutic groups. The estimated probabilities of survival were not significantly different among the 3 groups [P = 0.112]. BSO showed promising prophylactic and therapeutic effects on murine toxoplasmosis

Antiparasitic activity of cystine protease inhibitor E-64 against giardia lamblia excystation in vitro and in vivo

In this work, the therapeutic effect of E-64, a broad spectrum cysteine protease inhibitor against Giardia lamblia excystation was studied in vitro and in vivo. Purification of cysts from heavily infected human faecal samples followed by excystation and axenic cultivation of the emerging trophozoites in TYI-S-33 medium were done. In vivo, the response was evaluated experimentally through counting oocysts out-put every other day until the infection eradicated from the stools of infected E-64 treated mice compared to untreated. Also, the histopathological examination of the small intestine was compared between both of the infected groups. In the present study G. lamblia cysts incubated with E 64 in vitro completely failed in excystation in 90% while trophozoites released on 10% [partially excysted on 5% and completely excysted on 5%] compared to 90% completely excysted on other non incubated [without E-64] of cysts beside, the trophozoites didn't release on 10% [partially excysted on 5% and completely non-excysted on 5%]. In vivo, the evaluation of the therapeutic response proved that the decreasing in the oocysts out-put counting every other day until the infection eradicated from the stools of infected treated mice was very marked in comparison to untreated mice. The differences were statistically significant. The histopathological examination of the small intestine of infected non treated group proved that all the different pathological grades were found while in infected E-64 treated group, only grade I was detected. So, E-64 showed a good therapeutic effect which raises its use in the treatment of human giardiasis

Infectivity of trichomonas vaginalis pseudocysts inoculated intra-vaginally in mice

In addition to the trophozoite, pseudocyst is another morphological form which is recently identified among genitourinary trichomonads. Although, this pseudocyst is competent to divide, its role in Trichomonas life cycle has not yet been confirmed. In this study the ability of intra-vaginally inoculated T. vaginalis pseudocysts to induce trichomoniasis in infected mice was evaluated in comparison to the trophozoites. Pseudocysts formation was induced by using thermal-freezing cycle method. The infectivity of the pseudocysts was proved by the presence of T. vaginalis parasite in mice's vaginal washes inoculated in vitro. SEM proved that the pseudocysts withstood on vaginal tissue for 72 hours post infection without any morphological changes. Although the histopathological studies using H and E, PAS and cathepsin D stain proved that there were no differences could be found between trophozoites and pseudocysts in onset of infection, but the pseudocyst had higher infectivity and invasive effects than the trophozoite. So, T. vaginalis pseudocyst is an active form that can induce trichomoniasis

Relation of gall stone type with bile composition

The present study was planned to investigate the relation of gall stone type with bile composition. This study was carried out on 41 patients who underwent elective cholecystectomy for calcular cholecystitis. These patients were classified into three groups according to the chemical constituents of stones: group [1] comprised 25 patients with cholesterol gall stones, group [2] comprised 8 patients with mixed stones, and group [3] comprised 8 patients with pigment stones. In addition 8 subjects were included as a control group. Blood sample, gall bladder bile and stones were collected from every patient. Fasting blood glucose, bilirubin, albumin, AST, ALT and lipid profile were measured by the traditional colorimetric methods. At the same time plasma ascorbic acid was determined by dinitrophenyihydrazine thiourea copper sulphate [DTC] reagent. In addition serum insulin was measured by chemiluminescent method and leptin by enzyme immunoassay. Biliary bile acids were measured by HPLC while biliary cholesterol, phospholipids, calcium, bilirubin and total protein were determined by colorimetric methods The present study revealed that ages of patients with pigment stones were significantly higher while BMI was elevated in patients with cholesterol gall stones. Serum cholesterol, TG, LDL, insulin and leptin were significantly increased while HDL and serum ascorbic acid were significantly decreased in patients with cholesterol gall stones when compared to control group. In addition, there was significant positive correlation between leptin and BMI, insulin and biliary cholesterol. In patients with cholesterol gall stones there were significant increase of biliary protein, biliary cholesterol and decrease of biliary phospholipids, total and direct bilirubin when compared to control group. In patients with mixed and pigment gall stones, there were significant decrease of serum HDL, biliary phospholipid, total and direct biliary bilirubin beside significant increase of indirect bilirubin when compared to control group. Concerning biliamy bile acids there were significant decrease of primary bile acids conjugates [GCA, TCA, GCDCA, and TCDCA] and significant increase of secondary bile acids conjugates [GDCA, TDCA, GLCA, and TLCA] in all patient groups when compared to control group. Leptin was associated with increased risk of cholesterol gall stone formation while advance of age was associated with pigment gall stone. Hypovitaminosjs C may have a pivotal role in gall stone formation specially cholesterol type. Expand of secondary bile acid pool and contracted primary bile acid pool play an important role in various types of gall stones

Serum leptin and biochemical markers of bone remodeling in osteoporotic postmenopausal women before and after antiresorptive treatment

To assess changes in the leptin and biochemical markers of bone remodeling in osteoporotic postmenopausal before and after antiresorptive treatment. Gynecology and Obstetrics Departments, Faculty of Medicine, Mansoura University. This study included sixty three postmenopausal women. They had hone mineral density and T-score more than 2 SD below normal healthy premenopausal women. In addition twenty normal premenopausal women 'were included as a reference group. The postmenopausal osteoporotic women were subdivided into three groups according to treatment modalities: group I: hormone replacement therapy [HRT]. Group II: alendronate therapy, and group III: combined HRT and alendronate. Blood and urine samples were collected from all groups before and 12 months alter treatment. Serum osteocalcin and leptin were measured by enzyme immunoassay while urinary deoxpyridinoline [Dpyr] was measured by chemiluminescence method. Bone alkaline phosphatase was determined by using agarose gel ectrophoretic method. Serum N-terminal propeptide of type procollagen [PINP] and serum C-terminal telopeptide of type I collagen [ICTP] were measured by radioimmunoassay. Total and bone alkaline phosphatase, usteocalcmn, serum ICTP, deoxypyridinoline and leptin were significantly increased while hone mineral density, T-score, calcium, inorganic phosphorus and serum PINP were significantly decreased in postmenopausal women in comparison to premenopausal healthy women. Serum leptin, osteocalcin, serum ICTP and urinary deoxypyridinoline were significantly decreased while serum PINP and bone mineral density significantly increased after treatment with HRT. aicadronate or combined HRT and alendronate for 12 months. Urinary deoxypyridinoline [marker or of bone resorption] showed most significant change [-60%] followed by serum PINP [marker of bone formation] [53.5%] during antiresorptive treatment. Serum leptin showed significant positive correlation with body mass index. osteocalcin. ICTP and deoxypyridinoline and significant negative correlation with serum PINP. body mineral density and T-score while hone alkaline phosphatase showed no significant correlation with leptin-Leptin has a regulatory role for hone formation in postmenoPausal wonton and those with high bone turnover could profit more from antiresorptive treatment than women with low or normal bone turnover

Study of some risk factors in diabetic foot ulcer patients

Objectives: To study plasma homocysteine and lipoprotein [a] levels together with sum other conventional tisk factors for atherosclerosis in diabetic foot ulcer [DFU] patients in comparism to diabetic palicnts without foot ulceration

Subjects and Methods: The study included 66 patients with type 2 diabetes [37 with DFU and 29 without DFU] and 20 healthy normal subjects of matched age and sex. DFU patients were classified into neuropathic, ischemic or neuroischemic ulcer according to neuropathy disability score. Plasma levels of total homocysteine [tHcy] was measured by enzyme immunoassay while lipoprotein [a] was measured by immunoturbidimetric method. Serum folic acid and vitamin BIZ were measured by radioimmunoassay

Results: Fasting plasma homocysteine and lipoprotein [a] levels were elevated in diabetic patients compared to the control subjects. DFU patients had significantly higher levels of tHcy and Lp[a] in comparison to non-DFU patients. Serum folate was lower in diabetic patients in comparison to the control subjects. DFU patients had significantly lower levels of folic acid in comparison to non-DFU patients. Serum levels of vitamin BIZ showed non significant changes in DFU, non-DFU and control subjects. Urinary albumin excretion [UAE] was significantly higher in the DFU patients in comparison to non-DFU patients. Homocysteine concentration showed significant positive correlation with UAE and Lp[a] and significant negative correlation with serum folic acid

Conclusion: DFU patients had higher levels of homocysteine and lipoprotein [a] and lower levels of folic acid in comparison to diabetic patients without DFU. Hyperhomocysteinemia and hyperlipoproteinemia [a] may partially explain the increased risk of macrovascular complications in DFU patients

Neonatal screening program in Dakahlia: hypothyroidism in clinically suspected cases

In the line with the priorities of the national health services towards preventive medicine early diagnosis and treatment of CH is emphasized. The aim was to assess thyroid status in clinically suspected cases. Subjects and The present study included two groups, the first is a set of neonates [800] attending the general health units in some villages in Dakahlia complaining of prolonged jaundice, constipation, abdominal distention, chocking and/or umbilical hernia. Neonatal TSH levels in filter paper spots taken from heels were assessed for this group. The second group included 114 infants suspected clinically as congenital hypothyroidism by pediatricians. Serum TSH and T4 were estimated for this second group. In the first group 690 neonates [86.25%] had TSH less than 20 micro IU/ml, 88 neonates [11%] had TSH of between 20 and less than 40 micro IU/ml and 22 [2.75%] has TSH more than 40 micro IU/ml. All neonates [110] with TSH more than 20 micro IU/ml [13.75%] had to be recalled after two weeks for reevaluation by serum TSH and T4 levels estimation. Transient hypothyroidism were detected in 70 cases [8.8%] and their serum T4 and TSH levels two weeks after, were normal. Forty neonates [36.4%] were missed and did not respond to our call. Effect of storage on neonatal TSH levels were evaluated in blood spots collected on preserved filter papers. The results revealed that there were gradual increases in the missed cases as zero when estimated with prolonged period of preservation. The results of the second group: Ninety cases [79%] were euthyroid with normal serum TSH and serum T4,17 cases [15%] were with mild elevated serum TSH [7-20 micro IU/ml] and normal serum T4. 7 cases [6.0%] were with elevated serum TSH [more than 20 micro IU/ml] and low serum T4 [less than 96 nmol/L]. second samples, two weeks after the first revealed normalized serum TSH and T4 in 23/24 case. One case presented with prolonged jaundice was diagnosed as overt congenital hypothyroidism. Prolonged jaundice is the presenting complain among 53/114 infants referred by pediatricians as suspected CH [46.49%]. One case Out of 114 studied infants was detected as true CH, while no case of true CH was detected among the neonatal cases. Serum confirmation tests [TSH and T4 levels] should be done in any infant who is suspected to have congenital hypothyroidism. The study assures the importance of public education and awareness in contributing to the cost effectiveness of screening program using filter paper blood spotting and supports the necessity of the rapid assay of filter paper spots within few days in large common collecting center for screening

Hepatitis C virus infection in B-cell non-Hodgkin's lymphoma patients

To determine the prevalence of HCV infection in patients with non Hodgkin's lymphoma in Assiut University Hospital and Cancer Institute of South Egypt. Eighty unselected patients with B-cell non-Hodgkin's lymphoma were tested for the presence of Hepatitis C virus antibodies using Elisa immunoabsorbant assay. Two control groups were utilized for comparison; the first is a cohort of patients with non-haematological neoplasms; and the 2[nd] is a group of healthy blood donors. Patients with previous history of blood transfusion or liver disease were excluded from the study. Thirty-eight of the 80 B-cell non-Hodgkin's lymphoma patients [47%] tested positive for Hepatitis C virus antibodies. About 24 out of 100 [24%] and 22 out of 80 [27.5%] patients tested positive for hepatitis C virus antibodies in the first and 2nd control groups. The results of this study indicate a higher prevalence of Hepatitis C virus infection in Assiut patients with B-cell non-Hodgkin's lymphoma than in the control groups. The prevalence of Hepatitis C virus infection in the 2 control groups, in turn, seems to fall within the estimated high prevalence in the general population of our country

Production of safe green natural colorant from carrot leaves [Daucus carota L.]

The amount of agricultural wastes seems to be high and could be used to obtain some important products. Carrot wastes represent about 36% of the total crop. These wastes could be used to produce compounds of beneficial and economic values such as natural green coloring matters to be used in manufacturing food products. The results obtained ascertained the possibility of producing natural green colour from carrot [leaves] to be used in food industries. Carrot leaves chlorophyll content degraded to about 50% of the initial content after 1 week storage under refrigerated conditions at 5 degreeC in polyethylene bags. The loss reached to 92% at the end of storage [six weeks] under the same conditions. Gradual increase of pheophytin content occurred during the first 3 weeks of storage and gradually decreased until the end of storage. The best treatment for extraction of green colour from the carrot leaves was by hot water at 95 +/- 3 degree C for 3 minutes containing 0.5% MgO and 1% CaCo3. Natural green colour was obtained as powder using the freeze-drying method. Addition of natural green colour [powder] to some foods increased their ash content. An increase in mineral salts such as Ca, Mg, Fe, Na, Cu and Zn occurred, in addition to improving taste, appearance and overall acceptability of such enriched foods with this natural green colour

N-acetyl-beta-d glucosaminadase and retinol-binding protein versus albuminuria in insulin dependent diabetic children.

Markers of tubular dysfunction [N-acetyl-beta-D-glucosaminidase and retinol binding protein], markers of glomerular dysfunction [glomerular filtration rate, urinary albumin excretion, blood urea and serum creatinine]and blood pressure were estimated in 32 insulin dependent diabetic children [IDDM] [16 males and 16 females] aged from 7 to 15 years, who had negative reagent strip test for proteinuria and in 10 age and sex matched healthy control children [6 males and 4 females]. Ultrasonography of the kidneys and fundus examination for the detection of retinopathy were done for all cases. The results indicated that tubular dysfunction, as evidenced by RBP and NAG, is more sensitive and precedes glomerular dysfunction in insulin dependent diabetics. However, RBP is a convenient, more sensitive, specific and early diagnostic tool for the detection of tubular affection than NAC. Albuminuria and increased GFR are early markers of glomerular dysfunction preceding the elevations of blood urea and serum creatinine. A reduction in hyperfiltration or GFR with increased albumin excretion is a marker of progression of renal disease. Hypertension is a feature of diabetic nephropathy after the appearance of microalbuminuria. However, the role of dietary modification programs and blood pressure control in normalization of kidney function in IDDP remains to be evaluated

Thyroxin binding globulin [TBG] and prealbumin [TBPA] levels in healthy subjects

This study was carried out on 120 carefully selected normal healthy subjects [56 males and 49 females in addition to 15 newborns]. Their ages ranged from newborn to Geriatric. Estimation of plasma TBG and prealbumin by RIA and immunoturbidimetric methods respectively was performed. Plasma TBG levels were nearly constant in females up to the age of 7.5 years and then showed progressive decrease, together with that of the male group, till the age of 16 years and then showed a slight gradual increase till the age of 65 years. The highest levels of TBG were observed at birth and at the age of 65 years and the lowest one was at age group of 15-18 years [about two thirds the highest levels]. No significant sex difference was encountered in TBC levels among all age groups. Pre albumin levels were nearly constant till the age of 8 years in males and decrease slightly in females. In both sexes, a marked increase was noted from 8-14 years, then the increase was much slower till the age of 24 years. The level started to decline slightly from age 24 to 40 years and markedly till age of 65 years. The highest level of prealbumin was in age of 15-35 years which was two and half folds that obtained for newborns and old age [65 years]. Prealbumin was significantly higher in male than females at the age range 15-50 years. Significant negative correlation was demonstrated between plasma TBC and prealbumin levels in all age groups

Transeferrin and haptoglobin, age and sex variations

Immunoturbidimetric technique was used to measure serum transferrin and haptoglobin levels in 120 subjects representing different age groups starting from birth till 80 years. Both sexes were represented almost equaly Plasma transferrin levels showed a gradual rise from birth till the age of 15 years and then dropped sharply at the age of 20 years. A slight decrease was then observed starting from the age of 24 to reach its lowest level at the age of 65 years. The highest level of transferrin was observed at 15 years which was significantly higher than in older age groups. Serum haptoglobin was absent in the newborn and started to appear at the age of one month. Then it showed marked increase from age of one month to the age of 30 Years, remaining relatively constant between 30 and 40 years then gradually increased to reach the highest level at the age of 65 years which was significantly different from other age groups. No sex difference was shown for either transferrin or haptoglobin at all age groups. So-age variations need more focusing to standardize normal levels which requires large number of cases, since it is limited by the high cost of kits in the present study