Genetic analysis of rifampin resistance in M. tuberculosis

Drug resistant strains of M. tuberculosis pose serious public health problem in Egypt. Inadequate treatment regimens and inefficient laboratory diagnosis of drug-resistant strains are likely major contributors to these problems. Mutations of the rpoB associated with rifampin resistance were studied in 25 rifampin resistant clinical strains of Mycobacterium tuberculosis isolated in Egypt. Of the 25 resistant isolates, 17 had a mutation in the 81 bp region of the rpoB gene by single strand conformation polymorphism [SSCP] and DNA sequencing. Only two resistance patterns could be detected by SSCP analysis. Sequence analysis revealed that Ser[531] - Leu arose most frequently missense mutation [76%] followed by Asp[516] - Val [24%]. The sensitivity and specificity of the SSCP result were 100% compared with that of the DNA sequencing. These results suggest that SSCP is an efficacious method for predicting rifampin resistance and would reduce the time required for susceptibility testing from 4-8 weeks to few days and it is useful for rapid screening of rifampin resistance in susceptible and fully resistant isolates of M. tuberculosis

Oncogenic effect of some environmental pollutants and some possible protective agents

Objective: to study the effects of some environmental pollutants [lead, nitrate, noise stress and tobacco smoke] as well as a chemical substance [estrogenic oral contraceptive pills] on the incidence of malignant transformation among offspring rats through transplacental transfer from mother to fetus. The protective effect of Nigella sativa and vitamin A against the possible cellular transformation in the offspring rats was also studied

Subjects and Methods: five groups of female Sprague-Dawley rats [30 rats /group] were fed standard commercial diet. The first four groups were exposed to noise [100 decibel] and tobacco smoke for half an hour, three times weekly and received contraceptive pills daily. Group I received low doses of lead and nitrate, group I1 and group I11 were treated as group I but they were protected by adding to the diet crushed N. sativa to group I1 and vitamin A to group 111, while group IV received normal diet mixed with high doses of lead and nitrate. The fifth group served as untreated control. After three months no more contraceptive pills were added to the diet to prepare the female rats for meeting. After labour, mothers and their offspring were no more exposed to noise or tobacco smoke. The rats were sacrificed 45 days after labour, sera and tissues were saved for the biochemical and histopathological investigations

Setting: National Cancer Institute [NCI], Cairo University

Results: results showed an increase in the levels of mutant p53 protein, alpha-fetoprotein [AFP], ferritin and gamma glutamyl transferase [GGT] in the sera of both mothers and their offspring rats in the unprotected groups [I and IV] compared to the control group. On the other hand, the addition of N.sativa and vitamin A to the diet, led to a decrease in the levels of mutant p53, AFP and ferritin. Histopathological changes ranging from mild to severe dysplasia were found in both the mothers and their offspring rats in the two unprotected groups, while mild and moderate dysplasia were found to a less degree in the vitamin A protected group and disappeared from those fed N. sativa

Conclusion: the combined effects of the tested pollutants caused liver cells injury, ranging from mild to severe dysplasia, and raised the levels of serum mutant p53 protein, AFP, ferritin and GGT in both mothers and their offspring rats, which may lead on the long-term exposure to malignant transformation. On the other hand, N. sativa and vitamin A have a good protective effect which decreased both abnormal biochemical and histopathological changes

Value and limitations of polymerase chain reaction and diffrent serological markers in diagnosis of single and mixed acute viral hepatitis

Introduction: Hepatitis viruses are endemic in Egypt, Many studies were performed in order to study the pattern of acute viral hepatitis [AVH] and all of them were hospital-based. The frequency of viral agents causing hepatitis in the previous studies were conflecting. Aim: to evaluate the value and limitations of polymerase chain reaction [PCR] and different seromarkers in diagnosis of single and mixed AVH. Patients: This study was performed in Sallam village near Assiut city. Active surveillance was performed on 1400 cases using house to house survey and frequent visit to rural health unit. 105 patients with AVH 60 males and 45 females with age ranged from 2 months to 65 years were included in the study. All patients had full history, general and abdominal examination including ultrasound, liver funtion test and different seromarkers. Anti hepatitis A virus [anti-HAV] IgM and IgG, HBs Ag, anti HBc IgM anti HEV IgM and IgG and dithiothreitol [DTT] anti-HCV IgM, cytomegalovirus [CMV] and Epstein Barr virus [EBV] IgM antibodies. Reverse transcriptase polymerase chain reaction RT-PCR] were performed for HCV and HEV. HA V was the most frequent cause of AVH 34 [32.4%], followed by AVE 29 [27.6%], AHB in 9 [8.6%]. Acute hepatitis C [AHC] was diagnosed only in 3 cases [2.9%] one single and two mixed with E virus. Hepatitis C RWA may be suppressed in mixed viral infection as it occurred in two of our cases who had mixed C/E viruses. Two cases were diagnosed CMV and one case mixed CMV/EB V. 70 [66.7%,] had single viral infection, while 14 [13.3%] had mixed viral infection and 16 [15.2%] were negative for all viral markers and diagnosed as non A-E hepatitis. 19 cases had clinical and laboratory evidence of AVH however they had positive anti-HCV Ab six months prior to inclusion, so they were diagnosed as chronic HC. 6 cases were excluded from the study and diagnosed as drug and toxin induced hepatitis. PCR aid variant seromarkers have an important role in diagnosis of AVH, however they have many limitations particularly in acute hepatitis C and mixed infection

Anticardiolipin, autoantibodies, and cryoglobulin in patients with chronic hepatitis C and their relation to thrombocytopenia

Introduction: Chronic infection with hepatitis C virus [HCV] has been found to be associated with autoimmune phenomena and extra hepatic manifestations. HCV was implicated as a cause of the development of anti-phospholipid antibodies, leading to the suggestion that this virus may be involved in the aetiopathogenesis of the anti- phospholipid syndrome [APLS]. Aim: to study the association between anticardiolipin, autoantibodies and cryoglobulin with chronic hepatitis C infection and their relation to thrombocytopenia. Patients: Thirty three patients with chronic liver diseases of different stages were included in this study. Twenty seven of them were due to hepatitis C virus [HCV] and 6 were due to hepatitis B virus [HBV]. Their age ranged from 35 to 52 years. Twenty healthy subjects were included as controls In addition to clinical and ultrasonografic examination complete blood count, anticardiolipin [aCL] [IgG and 1gM isolypes], antinuclear antibody [ANA], antismooth muscle antibody [ASMA] antimitochondrial antibody [AMA] and cryoglobulin were done for both patient and control groups. The frequency of aCL antibodies of IgG and IgM isotypes were significantly more in patients with chronic HC infection [59.2% and 81.5 respectively] than that in controls [5% for each] [P < 0.001]. There was no significant relations between the presence of aCL antibodies and the presence of thrombocytopenia. There was no significant difference in the frequency of aCL between patients with chronic HC and chronic HB infection. The frequency of cryoglobidins was more in HC positive patients [63%] than HB positive patients [50%], but the difference was not significant. However ANA and AMA were significantly more in patients with chronic HC group than patients with chronic HB group [P < 0.05] for each. A significant proportion of HCV patients had detectable aCL However no significant association between aCL antibodies and thrombocytopenia was detected

Comparison of the circulating carbohydrate antigen and the anti-SWAP antibodies in patients with active schistosoma haematobium infection

Twenty five patients, with age ranged from 5-25 years [mean +/- SD: 12.8 +/- 4.7] with active S. haematobium infection were investigated in the present study. They were subjected to complete clinical history, examination, abdominal ultrasonography and urine and stool examination. Nuclepore filtration method for quantitation of S - haematobium ova was performed. Immunological assays to study the levels of circulating carbohydrate antigen lusing monoclonal antibody 128 C3/3/21] and anti-SWAP [soluble adult worm antigen preparation] immunoglobulins IgG1, IgG4 and IgE were performed. The patients were re-evaluated again at 1, 3 and 6 months of receiving praziquantel therapy. It was found that all Patients had significantly high circulating carbohydrate antigen in their sera i.e. above the cut-off value. Also, there was a significant drop in the mean circulating antigen levels at 1, 3 and 6 months after praziquantel treatment. However, the levels of the circulating antigen did not reach to the cut-off value in, 12 patients 6 months after treatment; 5 of these patients had evidence of reinfection and/or persistence of infection. On the other hand, all the patients had positive ELISA reaction for IgGl and IgG4, while 5 patients had negative reaction for IgE through the different visits before and after treatment. The decrease in the mean levels of IgGl and IgG4 was statistically significant only after 6 months of treatment; but the mean levels of IgE showed significant drop at 3 and 6 months after treatment. These findings suggested that the detection of circulating carbohydrate antigen is more accurate in diagnosis of active infection, reinfection-and in evaluating the efficacy of treatment. We found a significant correlation between the circulating carbohydrate antigen levels and the anti-SWAP IgE during the active infection but no significant correlation was found between the antigen levels and IgGl and IgG4. This may be attributed to the fact that IgE antibodies to S- haematobium adult worm antigen increase with the parasite load. Further evaluation must be performed on a large number of patients with more prolonged period of follow-up

Species and infectivity of schistosoma snails in middle Egypt

This study was carried out, in order to determine, the species of Schistosome snails and its infection rate, in three villages around Assiut. Different Schistosome snails, were classified according to the size and age. The total number of Bulinus trancatus snails collected from the three villages was 4169, and there were 2 peaks of increased number of snails. Two hundred and six Biomphalaria alex and rina snails were collected from two villages only. None of the collected Biomphalaria alex and rina snails were found to be infected, and the infection was rates of Bulinus trancatus in the three villages were 0.06%, 0.12% and 0.4%. Each snail infected with Schistosoma haematobium or Echinostome cercariae was not found to be infected with any other trematode cercariae. The appearance of Biomphalaria alex and rina snails, together with decrease in the Bulinus/Biomphalaria ratio, as noted in this study, is most probably due to favorable environmental conditions brought up after the construction of the High-Dam

Ophthalmologic studies under the effect of oral contraceptives

Eleven cases of short-term Anorlar 1 users [age 20-30 years] and 22 of the long-term pill users [up to four years] with an age ranging from 30 to 40 years were examined in the Ophthalmology department for ritual acuity, tension of the eye and fundus examination. All cases showed normal tension of the eye and no pathological findings were found in the fundus pictures of these pill users, even up to four years of intake. The defects in visual acuity was not related to the pill intake as they could be explained by the errors of refraction

Study of blood lipids under the effect of oral contraceptives

Serum cholesterol and beta lipoproteins were studied after the intake of Anovlar and Anovlar 1 tablets for short duration [six months] and long duration [2 to 10 years]. There was a significant increase in total serum cholesterol and in beta [atherogenic] lipoproteins, more marked in the long-term users. The state of hyperlipaemia induced by the combined pills can play a role in the pathogenesis of venous thrombosis, either by accelerating clotting or by inhibiting fibrinolysis

Study of liver function tests under the effect of oral contraceptives

A battary of liver function tests were done for a group of normal [30 cases]. Anovlar 1 tablets were given for 20 cases and the tests repeated and there was a significant increase in serum transaminases, zinc and thymol turbidity tests but no change was found in the serum alkaline phosphatase.. A group of long-term users [30 women] were also investigated for this liver functions and there wrre no significant changes in the values of these tests, this reversibility of the liver function could be explained by the phenomenon of "adaptation"

Study of the effect of oral contraceptives on blood coagulation factors

Blood coagulation factors, I, II, VII, VIII and X were assayed before and after the intake of Anovlar 1 tablets for six months and also after the long-term use of Anovlar and Anovlar 1 tables for 2 to 10 year. In both group there was a significant increase in Factors I, II, VII and X. Factors VIII [antihaemphilia globulin] showed no significant change in the studied groups

Electro-gardiographic studies under the effect of combined pills

Seven cases of short-term pill users and 13 of long-term users of combined oestrogen-progestogen combination were studied by ECG tracings. There was no pathological changes in the ECG for the studied cases even after ten years of intake of the pills. This excludes abnormalities in the vascular, myocardial and conduction system of the heart

Fibrinolytic activity under the effect of oral contraceptives

Twenty women were given Anovlar 1 tablets cyclically for six months, and the fibrinolytic activity of their plasma was studied by the euglobulin clot lysis time. Pre-intake study was done for 30 women, and for another group of long-term users of Anovlar and Anovlar 1 tablets for 2-10 years. There was a significant shortening in the euglobulin lysis time in the two groups of pill users. This enhanced fibrinolytic activity counter balances the increase in the blood coagulation and thus preventing the occurrence of intravascular thrombosis