Ultrastructural study on biomphalaria alexandrina haemocytes infected with schistosoma mansoni in Egypt and its correlation with nitric oxide level

Some snails of Biomphalaria alexandrina can resist the infection of Schistosoma mansoni so this study aimed to clearly this mechanism by using light and electron microscopy [EM] and determine the role of Nitric oxide in this mechanism. B. alexandrina snails used in this study were exposed individually to S. mansoni infection according to their response they were classified into susceptible group [shed cercariae] and resistant group [failed to shed cercariae]. Snails not exposed to infection were included in this study as control group. Nitric oxide [NO] level was assayed directly in the soluble fraction of B. alexandrina haemolymph supernatants collected from each group of B. alexandrina snails were subjected to NO assay by the Greiss reaction. The level of NO in haemolymph of infected snails was significantly increased [p<0.001] than both control and non infected snails groups, however, in non infected snails group had significantly [p<0.05] compared to control group. This study when correlated the changes recognized by EM with NO level the pro apoptotic effect of high level of NO on the haemocytes. Characterization and identification of cell shape of haemocytes in both haemolymph and tissue were examined by light and electron microscopy. Examination of B. alexandrina snail's haemocytes revealed three types of different cells classified according to their shape and granular contents. These cells are granulocytes, amoe-bocytes and hyalineocytes. Electron microscope Study also revealed the important role of granulocytes and amoebocytes as defense mechanism against snail infection. NO is considered an important anti parasite molecule; intra-molluscan stages of parasites switch off host NO defense response

White and grey matter abnormalities in some neurometabolic disorders: a clinico-radiological correlation

Inborn errors of metabolism [IEM] are common throughout the Middle East, presumably because of the relatively high rates of consanguinity. Disorders of inborn errors of metabolism that affect the CNS are divided according to MRI picture into disorders involving grey or white matter or both. Only very few studies on inborn errors of metabolism have been done in Egypt. However, with the introduction of MRI and increasing awareness of the disease, more and more cases of metabolic diseases are being diagnosed. Many of these diseases if detected early can be cured and MRI might play a vital role in its early diagnosis. MRI has virtually replaced all other imaging modalities including CT in the evaluation of white matter diseases due to its excellent gray-white matter resolution and multiplanar imaging capability. The aim of this work is to identify the pattern of white and grey matter involvement in some neurometabolic diseases and to evaluate the role of MRI as an investigative modality in inborn errors of metabolism and to document its value in early diagnosis and hence management. This is a descriptive study done among 800 patients suspected of having an inborn error of metabolism and referred to the clinic of inherited metabolic disease at Cairo University Children Hospitals, at the centre of Social and Preventive Medicine. Only fourteen patients were found to have the diagnosis of metabolic disease and abnormal findings on MRI. Out of 14 cases, 11 were males [78.6%] while 3 [21.4%] were females. At presentation, the mean age +/- SD of the patients was 4.36 +/- 3.775 years, and their ages ranged from 9 months to 16 years. Four cases [28.5%] were diagnosed as Leigh disease, 4 cases [28.5%] as metachromatic leukodystrophy, 2 cases [14.2%] as Canavan disease, 1 [7.1%] as methylmalonic academia, 1 [7.1%] as Vander Knappe disease, 1 [7.1%] as mucolipidosis, and one [7.1%] as Wilson disease. Clinical presentation included positive consanguinity in 9 cases [64.3%], other sibs affected in 5 cases [35.7%]. Global developmental delay in 3 cases [21.4%], and developmental regression in 11 cases [78.6%]. Attacks of disturbed conscious level with metabolic acidosis were encountered in 5 cases [35.7%]. Seizures were encountered in 6 patients [42.9%] [four of them were generalized tonic clonic, one was multifocal clonic and one was hemifocal]. All types achieved good control on one or two antiepileptic drugs. Extrapyramidal symptoms were present in 5 cases [35.7%], and ataxia in 2 cases [14.3%]. On examination there was spasticity in 11 cases [78.5%], while only 3 patients [21.4%] were hypotonic. On measuring the skull circumference, 6 patients [42.9%] had microcephaly, 4 patients [28.5%] were macrocephalic and 4 cases had normal skull circumference. Fundus examination was done and revealed optic atrophy in 2 cases [14.3%]. MRI was a guide in reaching a proper diagnosis in these cases, definitive diagnosis was done by tandem mass spectrometry [TMS], enzymatic essay and skin or gingival biopsies according to each case. When properly used, MRI pattern recognition approach in conjunction with the other imaging tools can be very useful for separating the complex group of metabolic disorders into more manageable groups. Indeed, sometimes this approach allows a specific diagnosis to be made. Although the pattern of disease in MRI is not often specific, the distribution may suggest a differential diagnosis that may be pursued by increasingly specific enzyme analysis

Impact of cysteine protease inhibitor on infection of biomphalaria Alexandria with schistosoma mansoni and evaluation of protease activity in snail's haemolymph

This work aimed to study the effect of cysteine protease inhibitor on Schistosoma mansoni miracidia and their host Biomphalaria Alexandrina snails. The survival of miracidia decreased steadily with the increase in inhibitor concentration and the rising in their mortality was observed by increasing the inhibitor concentration up to 100 micro/ml. Exposure of snails to two concentrations of the cysteine protease inhibitor25 and 50 micro g/ml during miracidial exposure, reduced infection rate of the snails in the two groups 13.3% and 12.5%, respectively in comparison with its control group being 73.0%, Continuous exposure of the two concentrations of cysteine proteuse inhibitor in the two sizes [3 +/- 1mm] and [6 +/- 1mm], reduced infection rate being 3.03% and 5.5%, respectively. The control group of snails size 3 +/- 1mm was more susceptible to infection than that of the large ones [6 +/- mm], the infection rates was 73.0% and 15.6%, respectively. The protease activity in infected experimental groups increased significantly when compared to the un-infected control group [6.13 +/- 4.35] audit was also increased significantly in the one day tested group, compared to the other groups, in the majority of the infected groups

Immunolocalization of schistosoma mansoni and schistosoma haematobium antigens reacting with their Egyptian snail vectors

The reaction of the haemolymph and the tissue of infected intermediate hosts, Biomphalaria alexandrina and Bulinus truncatus to Schistosoma mansoni and S. haematobium antigens were investigated using the indirect immunoperoxidase technique. A new technique, Agarose cell block was used in collection of haemolymph which helped in collecting plenty of well formed cells in comparison to the ordinary one using the cytospin. Collected haemolymph and prepared tissues of uninfected and infected B. alexandria and B. truncatus were fixed and then reacted with anti-S. mansoni and anti-S. haematobium IgG polyclonal antibodies. The haemolymph and tissue of infected B. alexandrina and B. truncatus gave a positive peroxidase reaction represented by a brown colour. In haemolymph, the positive peroxidase reaction was detected mainly in the cytoplasm of the amoebocytes. In the tissue, it was detected in epithelial cells lining the tubules, male cells in the lumen of the tubules and in female oogonia cells along the periphery of the tubules. The similarity in the strength and distribution of positive reaction in B. alexandrina and B. truncates was observed as compared to control. Thus, the immunoperoxidase technique proved to be an effective indicator for the schistosome-antigen in the snails

Electrophoretic patterns of protein fractionations in hemolymph and tissues of Biomphalaria Alexandrina and bulinus truncatus during course of schistosome infection

Electrophoresis of plasma protein of B. alexandrina [uninfected and infected with S. mansoni] showed that the major dominant bands had molecular weights of 20,44,96,139 and 205KD in both types of snails. The 1day and 1 week post miracidia1 exposure [PME] groups were characterized by band 54 KD. All groups except a day PMA were characterized by a common band of MW 65 KD. Three days PME group had three bands of 123 KD, 150 and 177 KD, not found in other groups. The highest similarity index in 2 weeks PME and 5 weeks PME groups [during cercaria1 shedding] was 0.667 the lowest one was in 3-dys PME [0.5]. the 3-days PME had a unique band of MW 177.04 KD, not found in other groups. Similar electrophoretic pattern of B. alexandrina protein was seen. The major dominant bands had molecular weights of 14, 21, 80 and 140KD in both non-infected and infected snails. The 1day PME had a band of 48.483 KD, 3days PME had band of 87.985 KD, one-week PME group characterized by two band 61.761 KD and 70.33 KD. The two-weeks PME had a band 91.111 KD. While, the 5week PME [during cercaria1 production] was the only group that shared the common band of MW 115 KD with controls. The highest similarity index in 5 weeks PME [during cercaria1 shedding] group was 0.545 and the lowest one was in 1 week and 2 weeks PME [0.43]. The electrophoresis of plasma protein of B. truncates [uninfected and infected with S. haematobium] showed that the major dominant bands had molecular weights of 20, 30, 65, 80, 106, 117 and 170 KD in both type of snails. The 1 day PME group was characterized by three band of MWs 26.539, 51.891 and 91.509 KD. All experimental groups, except 5 weeks PME [during cercaria1 shedding] and control, had a common band of MW 45KD. Three days PME group had a characteristic band of 113.72 KD which was not found in any other group. The highest similarity index was in one week PME group was 0.857 and the lowest one in 1 days PME [0.5]. In B. truncatus tissue protein, the major dominant band by electro- phoretic pattern had molecular weights of 20, 45, 54, 80, 97, and 171KD in both type of snails. A day PME had of 73.544KD and a week PME had a band of MW 60.813 KD. Two and 5weeks PME groups had 2 bands of MWs 27 and 62 KD .All experimental groups had a characteristic band not found in control of MW 141 KD. The highest similarity index in 3-days PME was 0.8 and the lowest one was in 5 weeks PME during cercaria1 shedding [0.545].

Effect of double infection with schistosoma mansoni and echinostoma liei on some physiological parameters of BiomphalariaAlexandrina

The survival rate and fecundity of B. Alexandrina were greatly influenced when exposed to either S. mansoni or E. Liei miracidia. The snails exhibited much lower survival rate and fecundity when double exposed to both S. mansoni and E. liei miracidia than single exposure and control snails. The results indicated a disruption in the snail metabolism due to the exposure to S. Mansoni and E. Liei miracidia and this effect was more pronounced in case of double exposure to the two parasites. Protein concentrations in hemolymph and tissues were significantly reduced in all exposed snail groups than in the control group. A significant elevation in the levels of aspartate aminotransferase [ASAT] and alanine aminotransferase [Asat] enzymes was recorded in hemolymph and tissues of exposed snail than the unexposed control snails. The ASAT/ALAT ratios in tissue and hemolymph of single- exposed and unexposed B. Alexandrina did not exceed 1, while it increased up to 1.18 in hemolymph of double exposed snails. There were significant increases in the levels of acid and alkaline phosphatases enzymes in the exposed snails

Concentration of some trace element in the serum, hair and nail of alopecia areata patients

Fifty three subjects [41 patients with alopecia areata and 12 matched controls] were selected for this study. The alopecia group comprised 25 males and 16 females with age varying from 10 to 40 years. Thirty one patients had alopecia areata and 10 patients had alopecia totalis. Fifteen patients had nail changes. Zinc, iron, copper and magnesium were estimated in the serum samples of all individuals of the study [41 patients and 12 controls] and the hair and nail samples of 42 subjects [30 patients and 12 controls] by the use of atomic absorption spectrophotometry. No statistically significant differences were found between trace elements concentrations in patients compared to controls except for low hair and nail zinc. Also patient with nail changes have a significant nail zinc as compared to those nail changes. According to these results we can speculate that trace elements have no proven role in the pathogenesis alopecia areata except for zinc which may be involved in the pathogenesis of this disease