Endemic Diseases Institute

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    EVALUATION OF A NEW ELISA PANEL FOR SCREENING OF ATROPHIC GASTRITIS
    (University of Khartoum, 1996) Al abbas, Hatim ; M.Mukhtar, Moawia ; Osama, Laila
    Helicobacter pylori –associated atrophic gastritis is known to be a significant risk factor for gastric cancer. Among the well-known parameters of atrophic gastritis are the levels of serum gastrin-17and pepsinogin I&II, which are biomarkers of gastric antral and corpus mucosal activity, respectively. The aim of the study was to evaluate a new ELISA panel for screening of atrophic gastritis. This study was done in mafraq hospital in the U.A.E. A total of 60 dyspeptic out patients who underwent diagnostic upper gastrointestinal endoscopy with biopsy were recruited to the study. The levels of antibodies to helicobacter pylori and levels of pepsinogenI, pepsinogenII and gastrin-17 were measured using a new ELISA panel. The results of the ELISA panel were compared with result of histology the sensitivity and specificity of the test were calculated . A decrease in serum G-17 levels along with the antral atrophy were observed in patients positive to Helicobacter pylori ELISA test, the serum level of SPGI was also reduced in corpus atrophy irrespective of the presence or absence of the Helicobacter pylori infection. The diagnosis of atrophic gastritis obtained with the blood test panel of G-17, PGI&II and H. pylori antibodies is in a fair agreement with the biopsy findings. Therefore Gastro panel could be used as a tool for non-endoscopic screening of atrophic gastritis
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    EVALUATION OF A NEW ELISA PANEL FOR SCREENING OF ATROPHIC GASTRITIS
    (University of Khartoum, 1996) Al abbas, Hatim ; M.Mukhtar, Moawia
    Helicobacter pylori –associated atrophic gastritis is known to be a significant risk factor for gastric cancer. Among the well-known parameters of atrophic gastritis are the levels of serum gastrin-17and pepsinogin I&II, which are biomarkers of gastric antral and corpus mucosal activity, respectively. The aim of the study was to evaluate a new ELISA panel for screening of atrophic gastritis. This study was done in mafraq hospital in the U.A.E. A total of 60 dyspeptic out patients who underwent diagnostic upper gastrointestinal endoscopy with biopsy were recruited to the study. The levels of antibodies to helicobacter pylori and levels of pepsinogenI, pepsinogenII and gastrin-17 were measured using a new ELISA panel. The results of the ELISA panel were compared with result of histology the sensitivity and specificity of the test were calculated . A decrease in serum G-17 levels along with the antral atrophy were observed in patients positive to Helicobacter pylori ELISA test, the serum level of SPGI was also reduced in corpus atrophy irrespective of the presence or absence of the Helicobacter pylori infection. The diagnosis of atrophic gastritis obtained with the blood test panel of G-17, PGI&II and H. pylori antibodies is in a fair agreement with the biopsy findings. Therefore Gastro panel could be used as a tool for non-endoscopic screening of atrophic gastritis
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    Molecular characterization of Sudanese dermotropic Leishmania parasites
    (University of Khartoum, 2002) Imam, Mukhlid ; M. Mukhtar, Maowia ; M. Elamin, Dr.Elwaleed
    Leishmania parasites cause different clinical forms of leishmaniasis constituting major health problems in several countries. Starting from self-healing cutaneous lesions to the fatal, if not treated, visceral form. This study is aimed to characterize and cluster Sudanese dermal Leishmania isolates from different parts of the Sudan. Seventy samples of dermal lesions were collected representing 55 cutaneous ulcers and 15 lesions of PKDL. Of these a total of 57 parasites was successfully cultured, 47 from CL lesions and 10 PKDL lesions. The isolates were characterized based on their kDNA PCR-RFLP patterns and RAPD amplification profiles. PCR amplification of kDNA of Leishmania isolates resulted in amplification of 800 bp in 42 isolates and 700 bp amplicon in 15 isolates, corresponding to L.donovani complex isolates and L.major complex respectively. To investigate the genetic diversity of the isolates, kDNA PCR products were digested with Alu1 restriction enzyme, different RFLP patterns were detected, with noticeable diversity among isolates of the two complexes. Random amplified polymorphic DNA using OP A10 primer resulted amplification of diverse DNA fragments profiles. Phylogenetic trees based on kDNA /RFLP profiles of the two isolates clustered the isolates into two main clusters. Phylogenetic trees based on RAPD profiles of the isolates clustered the isolates into main 5 groups. No correlation between either kDNA /RFLP, or RAPD profiles and the clinical form were detected.
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    Leishmaniasis in the Sudan: Parasite Characterization and Phylogeny
    (University of khartoum, 2003) Mohamed, Elwaleed ; M. Mukhtar, Moawia
    The study concentrated mainly on characterization of Sudanese Leishmania isolates; and the phylogenetic relation between isolates from different clinical forms in addition to the relation between these isolates and reference leishmania strains. The study also included attempts to identify parasite factors associated with PKDL in Sudan. In order to achieve the mentioned aims, more than 120 Sudanese leishmania isolates were cultured from different clinical forms of the disease. Isolated parasites were primarily characterized by their isoenzyme profiles compared with reference strains.64 isolates were characterized using isoenzyme: 8 were identified as L.donovani, 9 as L.infantum, 44as L.archibaldi and 3 as L.major. Interestingly L.archibaldi was strongly associated with VL and was the main parasite isolated from PKDL. Total genomic DNA was extracted and kDNA PCR and genomic PCR were performed. The kDNA was performed using species-specific primers followed by restriction fragment length polymorphism (RFLP) with AluI restriction enzyme. The total genomic DNA was analysed using 2 sets of primers; RH1, RH2 and SG1, SG2. The first set was used to amplify parasite sequences by PCR while the second set was used to amplify the gp63 genes then followed by RFLP and hybridisation technique. Radiolabeled probes were used to hybridize the digested genomic DNA in 2 ways; after gp63 amplification and restriction enzyme digestion, and after restriction enzyme digestion without gene amplification. Gp63 PCR analysis of selected 29 isolates identified 26 as L.archibaldi and 3 as L.major. Differential display technique was used for RNA in order to determine the differentially expressed candidate genes between VL/PKDL paired isolates. Cloning and sequencing were used to achieve full descriptions of these differentially expressed genes and northern blot was done to ensure that genes were not false expressed genes. The results obtained in this study suggest that L.archibaldi is the major cause of VL, PKDL and ML and also associated with CL infection. Another important finding in this study is that the PKDL infection is caused by the same parasite that caused the VL form. The high incidence of PKDL in Sudan might be L.archibaldi infection.
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    Immunoglobulin genes re-arrangement in Sudanese patients with leukaemias: a lineage marker.
    (University of khartoum, 2003) Satti Awad, Manal ; Awad, Eltahir
    In the recent years major advances was carried out in the improvement in the quality of life and survival for patients with leukaemia especially childhood acute lympholastic leukaemia. These advances have been attributed to improvement in supportive care and identification of prognostic factors that helped to categorize patients into favorable prognostic groups that respond to routine chemotherapeutic agents and non-responding group that need intensive treatment and probably bone marrow transplantation. An important prognostic factor that helped to classify patients is identification of the lineage of the leukaemia. Myeloid and lymphoid cells are treated differently by drugs types and duration. Although most of the classification systems of leukaemias depend on morphology of the cell, cytochemsitry, immunophenotyping and cytogenetics which are commonly used in addition to morphology to classify patients. In Sudan the only available method of lineage characterization is morphological identification of the type of cells. The objective of this study was to determine presence of light chain immunoglobulin rearrangement in Sudanese patients with different leukaemias and to show that morphological diagnosis was relatively in-accurate. This was the first time to conduct this type of moleculobiological study in the Sudan for diagnosis of leukaemia. Forty eight patients with morphologically classify leukaemias consented to participate in this study. Demographic and clinical data showed that there was no significant gender difference in different morphological types of leukaemias. ALL leukaemias were common in the young age group, compared to AML and CLL. DNA samples from twenty four patients were analysed using primers for κ light chain gene rearrangement (VKI & VKII). Two samples (2/2) from AML patients showed rearrangement in VKI, but no rearrangement in VKII, while seven samples (7/11) from ALL patients had rearrangement in VKI and no rearrangement in VKII. In the chronic leukaemias 2 patients (2/2) with CLL and 6 patients (6/9) with CML showed rearrangement in VKI, but no sample showed rearrangement in VKII
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    Leishmaniasis in the Sudan: Parasite Characterization and Phylogeny
    (University of Khartoum, 2003)
    ntrated mainly on characterization of Sudanese Leishmania isolates; and the phylogenetic relation between isolates from different clinical forms in addition to the relation between these isolates and reference leishmania strains. The study also included attempts to identify parasite factors associated with PKDL in Sudan. In order to achieve the mentioned aims, more than 120 Sudanese leishmania isolates were cultured from different clinical forms of the disease. Isolated parasites were primarily characterized by their isoenzyme profiles compared with reference strains.64 isolates were characterized using isoenzyme: 8 were identified as L.donovani, 9 as L.infantum, 44as L.archibaldi and 3 as L.major. Interestingly L.archibaldi was strongly associated with VL and was the main parasite isolated from PKDL. Total genomic DNA was extracted and kDNA PCR and genomic PCR were performed. The kDNA was performed using species-specific primers followed by restriction fragment length polymorphism (RFLP) with AluI restriction enzyme. The total genomic DNA was analysed using 2 sets of primers; RH1, RH2 and SG1, SG2. The first set was used to amplify parasite sequences by PCR while the second set was used to amplify the gp63 genes then followed by RFLP and hybridisation technique. Radiolabeled probes were used to hybridize the digested genomic DNA in 2 ways; after gp63 amplification and restriction enzyme digestion, and after restriction enzyme digestion without gene amplification. Gp63 PCR analysis of selected 29 isolates identified 26 as L.archibaldi and 3 as L.major. Differential display technique was used for RNA in order to determine the differentially expressed candidate genes between VL/PKDL paired isolates. Cloning and sequencing were used to achieve full descriptions of these differentially expressed genes and northern blot was done to ensure that genes were not false expressed genes. The results obtained in this study suggest that L.archibaldi is the major cause of VL, PKDL and ML and also associated with CL infection. Another important finding in this study is that the PKDL infection is caused by the same parasite that caused the VL form. The high incidence of PKDL in Sudan might be L.archibaldi infection.
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    Prevalence of Hepatitis C virus antibodies in Khartoum State
    (uofk, 2003-01) Ahmed, Omer ; Elkhidir, Isam
    Hepatitis C is a blood borne liver disease caused by hepatitis C virus (HCV). First identified in 1989, the disease was initially known as “Non-A, Non-B Hepatitis”. The hepatitis C virus belongs to the Flaviviridae family of viruses, and spread primarily through direct contact with the blood or body fluids of infected individuals. With the increasing use of antibody testing, the recognized prevalence of HCV infection has increased and an estimated 3% of the world’s population currently infected, equating to 170 million chronic HCV carriers world wide. HCV infection is a leading cause of chronic liver disease, including cirrhosis of the liver; therefore World Health Organization (WHO) recognizes hepatitis C as a global health problem. The aim of the study is to determine the prevalence of hepatitis C virus antibodies in Khartoum State, and to determine the possible modes of transmission. To achieve the objectives of the study antibody to HCV (anti- HCV) was studied by third generation Enzyme Linked Immunosorbent Assay (ELISA) 201 blood donors, 158 pregnant women, 198 dialysis patients, and 18 multiple transfused patients. According to the result obtained, anti- HCV was present in 0.4% among blood donors, and 0% in pregnant women. It was found in 28% dialysis patients and 5.5% among multiple transfused patients. Thus HCV infection was found predominantly among dialysis patients, and the possible risk factors were blood transfusion, longer duration, and loose application of universal precautions. Also this study concluded that dialysis especially in countries with high prevalence of HCV may be with possible mean of transmission of HCV. According to the results obtained, the study suggests some recommendations.
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    Cytogenetic And Fish Analyses In Sudanese Patients With Dysmorphic Features, Ambiguous Genitalia, And Infertility
    (University of khartoum, 2004) MOHAMED, MONA
    Over the past few decades, cytogenetic analyses have come to play an important role in the diagnosis, clinical management, and prognosis of several human diseases especially those associated with numerical and/or structural chromosomal changes, leukaemia, lymphomas, and soft and bone tissue tumors. The present study presents the first investigation in Sudan using cytogenetic and molecular cytogenetic techniques.
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    MOLECULAR CHARACTERIZATION AND DRUG RESISTANCE PATTERNS OF MYCOBACTERIUM TUBERCULOSIS ISOLATES FROM KHARTOUM STATE
    (University of Khartoum, 2004) Mukhtar, Moawia M. ; ELHASSAN, ALI MOHAMMED
    The present study was an attempt to study the epidemiology of pulmonary tuberculosis, characterization and identification of the causative mycobacteria and determination of their susceptibility to rifampin as a marker for drug resistance in the Khartoum state, Sudan. One hundred and fifty patients were consented and selected for the study. All sputum samples examined showed the characteristic AFB appearance of serpentine cords. All smear positive samples showed colonies of growth characteristic for Mycobacterium tuberculosis and all of them showed the presence of AFB on indirect smearing. Demographic data of consented patients were obtained from 147 patients out of 150 patients. All age groups were found to be affected by tuberculosis with varying frequencies. Five patients (3.4%) from group one (5-10), 32 (21.8%) from group two (11-20), 43 (29.3%) from group three (21-30), 30 (21.8%) from group four (31-40), 19 (12.9%) from group five (41-50), 8 (5.4%) from group six (51-60), 6 (4.1%) from group seven (61-70), and 2 patients in group eight (1.4%). 98 (66.7%) of the enrolled patients were males while 49 (33.3%) were females. The disease was found to be prevalent nearly in all provinces and localities of Khartoum state. 54 (36.7%) patients were found to be resident in the different localities of Khartoum, 56 (38.1%), and 27 (18.4%) were found to live in Khartoum North or Omdurman respectively, while 10 (6.8%)patients were considered as non Khartoum residents. 77 (52.4%) patients were singles (59 males, 18 females), 68 (46.3%) were married (35 males, 33 females), while one (0.7%) was scored for each of the separated and divorced categories. 39 (26.5%) were students, 30 (20.4%) were labors, 34 (23.1%) were housewives, 31 (21.1%) were business persons, 5 (3.4%) were employees (2 officers, 2 teachers, and 1 policeman), 5 (3.4%) were farmers, and 3 (2.1%) were homeless. 45 Sudanese tribes were recorded in this study. Patients from Denka, Gaalin and Nuba tribes were found to be significantly more than those from other tribes. The cough was the most common complaint (143, 97.3%), while other symptoms percentages were as follows, fever (121, 82.3%), loss of weight (96, 65.3%), shortness of breath (86, 58.5%), and haemoptysis (13, 8.8%). On duplex PCR, 129 out of 135 clinical isolates investigated by molecular biology techniques showed 235 bp DNA amplicons, indicating that they were M. tuberculosis complex strains while six isolates showed no DNA amplicons at neither 235 bp nor 136 bp. On PCR restriction enzyme analysis of rpoB gene (351 bp) the conserved rpoB gene present in all mycobacteria was amplified in 134 of our clinical isolates examined so far in this investigation with the exception of only one isolate which showed no amplified DNA bands. On nested PCR based sequence analysis amplifications of the rpoB gene of the clinical isolates resulted in the amplification of the target gene in all the examined isolates (133), at205 bp and 157 bp respectively, confirming that all of them were M. tuberculosis species. The G+C% content of the obtained nucleotide sequences was determined. A mean of 68.56% of G+C% content was determined. On analysis of the detected rpoB gene mutations, genetic alterations within the 81 bp hot spot region of rpoB gene known to be associated with rifampicin resistance were detected in eleven (11/133, 8.27%) of the studied isolates, of which the mutation Ser531 → Leu was the more common one (6, 54.55%), while mutations Leu511 → Pro, Asp516 → Gly, Asp516 → Val, Leu521 → Leu, and His526 → Tyr occurred once with the rate of 9.09% for each
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    Leishmania donovani in Eastern Sudan an innovative experimental approach for parasite cultivation and in vitro drug sensitivity testing
    (University of Khartoum, 2005) Elamin, Hayam ; Hassan Sharief, Abdalla
    From the early 1900s, visceral leishmaniasis (VL) has been among the most important health problems in Sudan, particularly in the main endemic areas in the eastern and central regions. Several major epidemics have occurred, the most severe--in Western Upper Nile province in southern Sudan, detected in 1988-- claiming over 100,000 lives. The disease spread to other areas that were previously not known to be endemic for VL. Major upsurges in the number of cases were noted in the endemic area. These events triggered renewed interest in the disease. In this study we described a relatively simple medium formula using common, available and inexpensive ingredients that can be used in place of medium that requires fetal calf serum enhancement for promastigote forms. This medium supported the growth of parasites at rate comparable with those obtained with serum supplemented RPMI-1640. Parasites grown in this medium shown moderate to high infectivity rates when being inoculated to pass through macrophage cell line (about 27- 84% of macrophage where infected) and no difference was observed in their infectivity and replication potentials inside cells (for both responsive and unresponsive parasites) between these parasites and that ones grown in parallel in RPMI-1640. Six L donvani. Isolated and one L. major was in vitro tested to Pentostam and Amphotericin B in their amastigote form (using J774 murine macrophage like cell line and as axinecally grown amastigote). The number of parasite in the infected decreased steadily at drug concentration 7.8μg/ ml to 250μg/ ml (in Pentostam) and 0.22μg/ml to 0.75μg/ml (in Amphotericin B) and the capacity of parasites to replicate was also affected. L.major showed the same response as other tested strains. In conclusion: The use of new CML medium can easily take the place of NNN or any other medium, because of it easy to prepare and used instantly, economic, available when needed, also the shelf life is about 30- 45 days. The fact that this medium is similar to other culture media as far as durability and quantity of produced parasites are concerned might give it an advantage over the others currently used. We also showed that the in vitro drug sensitivities did not match clinical unresponsiveness in the six L.donvani isolates, and the in vivo unresponsiveness does not necessarily mean primary parasite resistance. Active and dividing populations of axenically cultured amastigotes were generally more susceptible to Pentostam and Amphotericin B than their corresponding promastigotes. All the tested isolates were found to be highly susceptible to amphotericin B, that mean Amphotericin B is suitable second line drug if patients can tolerate it is toxicity
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    Characterization of Leishmania amastigote and axenic form antigens
    (University of Khartoum, 2005) M. Mukhtar, Maowia ; Awad, Aymen
    This study was designed to identify amastigote and axenic form antigens of leishmania major and leishmania donovani isolates. Two in vitro methods were used to transform the parasite isolates from promastigote to amastigotes and amastigote like (axenic) form. The first was in vitro infection of macrophages cell line J774 with leishmania promastigote, at 37o C with 5% CO2. The second was the culture of promastigote at 37o C with low pH (5.5), and 5-10% CO2, for axenic form transformation. Proteins were extracted from promastigote, amastigote, axenic form and J774 macrophages, simultaneously infected with L.major and L.donovani. Proteins were fractionated using 12% SDS PAGE. Antigens were detected using both dot blot & western blot techniques. PCR method was performed for detection of leishmania parasites in infected J774 macrophages. L. major was faster in infectivity of macrophages cell line than L. donovani (duration time for L. major 3-5 days, for L. donovani 7-9 days). Shared protein bands ranging from 26-116 kDa were detected by SDS PAGE in all stages. Dot blot technique revealed positive results, while western blot detected a unique antigen band of 16 kDa in amastigote from infected macrophages. PCR results were positive for both isolates showing that co infection is possible, and no signs of genetic recombination at kDNA were detected in macrophages simultaneously infected by L.major and L.donovani.
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    Local and systemic immune response of a combined anthrax and sheep pox vaccine
    ( 2005) MOHAMED, MOAWIA ; MOHAMED, ABDELRAHIM
    This study wascarried out to produce a combine anthrax and sheep pox vaccine and to compare its immune response with each of the single sheep pox and anthrax vaccine. Anthrax vaccine was produced in Roux flasks as a live spore vaccine, while sheep pox vaccine (SPV) was produced on lamb testicle (LT) monolayers. The combined vaccine was made from mechanical mixture of Bacillus anthracis (Sterne strain) live spores and the concentrated sheep pox vaccine, the mixture was lyophilized in 5 ml vials. The dose of each component of the final product was 1.0x 109 live spores/ml and 2.5log10 TCID50/ml of anthrax and sheep pox vaccine respectively. The potency of the combined vaccine was done by counting the number of the viable spores and by titration of sheep pox virus on sheep flank region. The viable spores count was found unchanged at the time vaccination (6 months of preservation at – 200 C, the virus produced nodular lesions on sheep flank when given intradermally. When the end point titration of sheep pox virus was conducted on flank region, the number of positive percentage was found 301and 277 for nonvaccinated control, 84 and 25 for sheep vaccinated with sheep pox vaccine and 50 and 54 for sheep vaccinated with the combined vaccine.
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    The Immunological Characterization of Hyper-reactive Malarial Splenomegaly Syndrome (HMS) in Sudan
    (University of Khartoum, 2005) Abdel Hady, Tayseer ; Mohammed, Ibrahim
    The present work aied at studying the immunological characterization of Hyper-reactive Malarial Splenomegaly Syndrome in Sudan. A hospital based study was carried out in the period between January 2000 and December 2004 in Ommdurman Tropical Diseases Hospital. HMS patients were interviewed and they had many investigations to differentiate HMS from other diseases which cause splenomegaly. Thirty one samples were collected to determine the molecular characterizations of the malaria parasite and to determine immunological responses to malaria candidate Circumsporozoite protein (CSP), also to measure the level of total IgM in Sudanese HMS patients' sera and to measure their cytokines (IL-10 & IFNγ). Males were found more affected by the disease (61%male - 38%female). The most common age group affected was between eleven and thirty two. Microscopic detection has shown that only one HMS patient had positive blood film for P. falciparum. No other malaria parasite species were detected. Using PCR the result was in agreement with the microscopic examinations P. falciparum parasite was detected in one patient, and no infection with P. vivax was detected. IgG antibodies to malaria specific antigen (CSP) were measured in 31 sera samples, IgG titers were found to be high but not significant in HMS patients compared with positive malaria control, whereas positive malaria controls showed significantly higher titers compared with European negative malaria control (P=0.005).
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    IMMUNE RESPONSES TO BRUCELLA VACCINE AND GENETIC DIVERSITY OF SUDANESE CAMELS BREEDS
    (University of Khartoum, 2005) E.Ibrahim, Muntasir ; M. Mukhtar, Moawia ; Ismail, Layla
    Immunological and genetic studies were performed in Sudanese camels. The aims of the former were to determine the optimum concentration of Ficoll for separation of camel lymphocytes, and to measure the cellular immune response of Sudanese camels which were vaccinated with Brucella vaccine. The genetic studies were performed to analyze the genetic diversity and relationships between a five Sudanese camels breeds i.e. Anaffi, Arabi, Rashidi, Bishari, and Kenanni. The immunological results revealed that 10 % Ficoll concentration was efficient for separation of camel lymphocytes. The colorimetric assay which was used for measuring the degree of the proliferative response of lymphocytes to various stimulations revealed the mitogenic effect of the PHA on the camel lymphocytes. Also low stimulation indices were calculated from lymphocytes isolated from non vaccinated camels, whereas lymphocytes isolated from vaccinated camels showed high values of stimulation indices, indicating the high magnititute of cellular immunity produced in camel followed vaccination with Brucella vaccine. The genetic studies were performed in the above mentioned five Sudanese camel breeds. Genomic DNA was extracted from 128 individuals representing the five breeds. Two types of polymorphic DNAThe first type was Random Amplified Polymorphic DNA (RAPD) marker. RAPD analysis was performed by using Polymerase Chain Reaction (PCR) with one random primer to amplify the genomic DNA of different camel breeds. The primer resulted in amplification of 18 polymorphic bands in the five breeds ranged in size from 150 – 1700bp. The mean number of bands that were produced in the five breeds ranged from 5.3 – 6.9 in Anaffi and Kenanni breeds respectively. The average value of bands sharing within a breed ranged from 0.54 – 0.87 between Kenanni and Bishari individuals respectively. The values of average bands sharing between breeds ranged from 0.212 – 0.359 between Anaffi- Kenanni, and Arabi – Rashidi, respectively. The studies with eight labeled microastellite primers indicated that seven of these primers were found to be polymorphic in the five camel breeds. The eighth primer was polymorphic in four breed and monomorphic in Kenanni breed. The number of alleles generated by the eight primers in the five camel breeds ranged from 1 – 19 alleles per locus. The observed hetrozygosity ranged from 45 % - 53 % in Bishari and Anaffi respectively. Four of the microsatellite loci were found to show significant deviation from HWE in some breeds. When F statistics test was done to measure the genetic diversity among breeds, it was found that the value FIS, which measures inbreeding in subpopulation, was found to be 17.4 % . The value of FIT, which measures inbreeding in the total population, was 20.2 % , while the value of FST, which is an indication of differentiation between breeds, indicated that the five breeds were poorly differentiated with FST value of only 3.3 % .
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    HLA Frequency of HLA-DRB1&HLA-DQB1 alleles in Mycetoma patients
    (University of Khartoum, 2005) M. Mukhter, Moawia ; Ibrahim, Maha
    Mycetoma (maduromycosis), True fungi-eumycetoma is the common cause of black grain eumycetoma in Sudan. It includes madurella mycetomatis-cause about 70% - 75% of all mycetoma cases in Sudan. The present study, which is the first molecular report on the association of HLA class-II alleles with Mycetoma infection in Sudanese patients. The objective of this study was to determine the frequency of the different alleles of HLA class II (DRB1, DQA1 and DQB1) in eumycetoma infected patients of Sudanese genetic backgrounds in an effort to determine possible association between the prevalence of certain alleles that could be associated with the occurrance eumycetoma infection. A total of 53 eumycetoma patients and 55 healthy control individuals were selected to determine the frequency of HLA-DRB1& HLA-DQB1. Both HLA-DQB1 and HLA-DRB1 genotyping were performed using PCR and sequence-specific (SSP) reverse hybridization oligonucleotide probe and analyzed with the Lambda Key Typing System and Lambda software. HLA-DRB1*13 and HLA-DQB1*06 were significant present in eumycetoma patients respectively (OR=2.543& chi-Square test =0.028) (OR=4.552& chi-Square test =0.000), while HLA-DRB1*11 and DQB1*01 (OR=0.313& chi-Square test =0.016) (chi-Square test =0.001) were less prevalent in eumycetoma patients.
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    T helper cells1/ T helper cells2 conversion to T helper cells1 immune response switch: the case of post kala-azar dermal leishmaniasis
    ( 2005) Abd Elmonim, Salma
    The Leishmaniases are a group of diseases caused by the genus Leishmania of the family: Trypansomatidae. They are transmitted by the female sand flies of the genus phlebotomus in the Old World and lutzomyia in the New World. The clinical forms in man include visceral leishmaniasis (VL), post kala-azar dermal leshmaniasis (PKDL), diffuse cutaneous leishmaniasis (DCL), mucocutaneous (MCL), self-healing cutaneous (CL). VL is the most serious and is fatal if not treated (El- Hassan & El- Hassan, 1997; Zijlstra & El-Hassan, 2001a; Zijlstra et al., 2001b; Musa et al., 2002). VL is caused by Leishmania donovani in the Old World, L.infantum in the Mediterranean basin and L.chagasi in the New World. It is characterized by development of fever, splenomegaly, hepatomegaly, weight loss and pancytopenia. Often complications occur such as epistaxis or bleeding from other sites, and concurrent infections (Zijlstra & EL-Hassan, 2001a).
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    In vitro induction/augmentation of unresponsiveness to sodium stibogluconate in Leishmania donovani isolates
    (University of Khartoum, 2005-07) Hamad, Sara
    Visceral leishmaniasis is a major health problem in Sudan; recent reports highlighted the emergence of SSG-resistant strains. Thirty Leishmania donovani isolates were successfully grown and passed in increasing concentrations of sodium stibogluconate (SSG). The (IC50) and the (IC90) of the isolates were initially established and were found to be 0.4 ± 0.04 mg/ml and 0.8 ± 0.1 mg/ml respectively. Following successful induction of resistance in the isolates, the IC50 and the IC90 rose to 50 times and 35 folds higher than those of the wild type (before induction of resistance) respectively (p<0.05). The time and the number of passages required for the isolates to recover to early log phase following exposure to SSG were first increasing with increasing SSG concentrations and then began to decline despite continued increasing SSG concentrations. The isolates were characterized as L.donovani using kDNA-PCR. SDS-PAGE technique showed that the resistant isolates developed additional three bands in their protein band profiles. Random amplified polymorphic DNA (RAPD) analysis, using three arbitrary oligonucleotide primers confirmed that the wild isolates were genetically divergent with three primary genotypes. For each primer, the PCR-RAPD band patterns of the resistant isolates were found to be similar to each other but different from those of the wild isolates. The resistant isolates were closely similar in their band patterns to that of one of the primary genotypes of the wild isolates. In conclusion, in vitro resistance to SSG was successfully induced/augmented in L.donovani isolates as evidenced by the increase in IC50 and the IC90 of the strains following passages in increasing concentrations of SSG. The shortening of time and the number of passages for the isolates to recover following SSG pressure is an added proof of acquiring resistance to SSG. L.donovani isolates from Sudan are genetically diverse. Two possible explanations were presented to explain the uniformity of PCR-RAPD band patterns of the resistant isolates. Firstly, two or more different strains could be present from the start (SSG-sensitive/resistant), the drug sensitive one/s was eliminated by the drug, while the other recovered after removal of drug effect (genetic selection). Alternatively, the isolates changed their DNA (genetic mutation) to face the drug pressure. Further studies with large numbers of isolates using PCR-RAPD primers (A12, A13 & IL-0875) are needed.
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    Humoral Immune Response to Malaria Vaccine Candidates (RESA and CSP) in Malaria Endemic Areas in Sudan
    (University of Khartoum, 2005-07) M.Elhassan, Dr. Ibrahim ; BASHER, SARA
    The present study was carried-out in Kosti town, White Nile state, Sudan. This area is characterized by a seasonal but stable transmition. During two cross-sectional studies curried-out in March 2003 and January 2004, a total of 173 samples was collected to determine the parasitological and molecular characterization of the parasite and to determine the immune response to malaria vaccine candidates (RESA and CSP). Microscopic examination has revealed that the parasite point prevalence was 26.4% and 38.5% in 2003 and 2004 respectively, and 96.2% of all malaria cases were due to P.falciparum. Using PCR technique, 21 out of 173 (12.1%) (9 samples in 2003 and 12 samples in 2004) samples with negative BFs were found to harbour low parasitaemia, this lead to increase in PCR point prevalence. Using merozoite surface proteins (MSP1 and MSP2) as genotypic markers, 71 samples were successively genotyped for the presence of MSP-1 alleles (MAD20, K1 and RO33) and MSP-2 alleles (FC27 and IC1). The frequencies of these alleles indicated that RO33 is the predominant MSP-1 allelic family and IC1 was found to be the predominant allelic family for MSP-2. In this study 10 allelic sizes for MSP-1 and 10 allelic sizes for MSP-2 were found to be shared between samples collectedin 2003 and 2004 (3 sizes for MAD20, 3 sizes for K1, 4 sizes for RO33, 6 sizes for IC1 and 4 sizes for FC27). The mean number of parasite clones per individual infection was 3.25-3.5 in 2003 and 2004 respectively. 22.3% and 57.7% samples in 2003 and 2004 respectively harboured more than one parasite clone
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    MOLECULAR CHARACTERIZATION OF HUMAN PAPILLOMA VIRUS FROM CERVICAL PRECANCEROUS AND CANCER SPECIMENS AMONG ETHIOPIAN AND SUDANESE WOMEN
    (University of Khartoum, 2005-10) Abate Waktola, Ebba ; Elhassan, Ahmed M. ; El-Tayeb, Muntasir ; El-Hassan, Ibrahim ; Engers, Howard ; Aseffa, Abraham
    Cancer of the uterine cervix continues to be a serious public health threat of global importance; 273,500 women are estimated to be dying from it annually. According to WHO reports, the age adjusted incidence rate of cervical cancer in Ethiopia and Sudan are 35.5 and 19.02 per 100,000 population respectively. Molecular and epidemiological studies have shown that Human Papilloma virus (HPV) is considered as the etiological agent for cervical cancer. Designing an effective vaccine against oncogenic HPV genotypes could have immense impact on the global cervical cancer burden. It was shown that a person protected against a specific type of HPV would still be at risk of getting infected with other cancer-inducing HPV genotypes. This necessitates that individuals living in different geographical localities receive vaccines based on the specific genotypes prevalent in that particular area. A retrospective molecular analysis for Human Papilloma virus was done on 245 formalin fixed paraffin embedded cervical biopsy samples bearing different histopathologic abnormalities that were collected from Ethiopia and the Sudan (160 and 85 samples respectively). The median age of the women who had given the biopsy samples were 45 and 48 (Range: 21-85; 30-75) for samples collected from Ethiopia and the Sudan respectively. DNA was extracted using phenol-chloroform extraction method and PCR was done to amplify the house keeping gene, β-globin, with PCO4, GH20 primers. Institutional ethical clearance was obtained from AHRI/ALERT, Addis Ababa University and Institute of Endemic Diseases ethical review committees. Amplification of HPV and subsequent genotyping was done on samples that were positive for the housekeeping gene using SPF10 primers and Line Probe Assay (LiPA) respectively. Accordingly, 93% (149/160) and 94% (80/85) of samples collected from Ethiopia and Sudan respectively were positive for HPV DNA. High risk (HR-HPV) and Low risk (LR-HPV) HPV genotypes were identified from 93% (149/160) and 13% (21/160) of all samples collected from Ethiopia respectively. Among the samples collected from Sudan, 94% (80/85) harbored high risk and 11.7% (10/85) low risk HPV v genotypes. Low risk HPV genotypes were detected as part of a mixed infection with at least one other HR-HPV type in both groups. HPV 16 was found to be the most frequent HPV genotype in Ethiopia and the Sudan accounting for 91% (136/149) and 82.5 % (66/80) respectively. Next to HPV 16, HPV 52, 58 and 18 were the second, third and fourth common HPV genotypes identified in Ethiopia. On the other hand, HPV 18, 45 and 52 were the second, third and fourth HPV genotypes identified in samples collected from Sudan. Mixed infections mainly composed of HPV 16, 18, 31, 33, 35, 45, 52, 58, and 68 were observed from samples collected from the two countries. Multiple infection with more than one HPV genotypes were identified in 59% (88/149) samples positive for HPV genotypes from Ethiopia and among 49% (39/80) HPV positive samples from the Sudan. The trend of having multiple genotypes reached to its highest peak in the age group 41- 60 and declined after the age 60 in samples collected from the two countries. It is recommended that a wide population-based epidemiological study be conducted to define the exact picture of this disease. A suitable vaccine targeting mainly HPV 16, 18, 45, 52 and 58 will have substantial impact on cervical cancer control in the two countries.
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    Leishmania donovani In Southwest of Sudan: Rapid assessment and Application of Geographic Information System
    (University of Khartoum, 2006) M. El Magzoub, Ranyab ; Hassan, Abdalla
    Visceral leishmaniasis (VL) comprises a major health problem in several parts of the Sudan and it leads to severe morbidity and high mortality if uncontrolled. Western Upper Nile State and Southern Kordofan State were a scene of a huge outbreak that claimed hundreds of thousands of lives in the late eighties, which demonstrated the inability, and inadequacy of the health authorities, at that time, in dealing with such disasters that are more likely to occur from time to time. Control of VL is obstructing by the elusive nature of the vector and failure to identify a reservoir host. Case detection and treatment with antimonial drugs contribute to disease control. In this study we introduced the concept of rapid epidemiological assessment of patterns of L. donovani infection and visualized the data in a geographic information system (GIS) hoping to open new avenues into the question of VL control. Coupling this map with the map of vector distribution and climatic changes will provide valuable information that will help in disease control, despite our meagre health resources. This survey was based on simple in vivo and in vitro immunological techniques (leishmanin skin reactivity and the Direct Agglutination Tests), combined with clinical history to obtain data about the spectrum of L donovani infection in communities at risk of developing VL. The data was represented in map format to give an enhanced visual impact. Treatment and future control strategies can easily be formulated from such data. XI In this study clinical history and immunological tests were conducted on 1781 volunteers randomly selected from the mesairiya tribe who areliving in western Kordofan State. The mean age of the selected volunteers was 20.7 years with an equal male:female ratio. The overall leishmanin skin reactivity of ≥5 was 24.5%, where children below 15 years had higher (13.2%) leishmanin reactivity compared to 11.3% among adults above that age. The DAT results showed that 88% (1567/1781) of the study volunteers had DAT titres below the cut-off point of 3200, whereas 6% (112/1781) had a DAT titers = 3200, while the rest 5.7% (238/1781) had titres > 6400. DAT results were shown in Table (3.5) and Figure (3.7). Six L donvani and one L. major strains were in vitro tested in their amastigote forms against Pentostam and Amphotericin B using J774 murine macrophage-like cell line. The number of parasite in the infected cells decreased steadily at drug concentration 7.8μg/ml to 250μg/ml for Pentostam and 0.22μg/ml to 0.75μg/ml for Amphotericin B and the capacity of parasites to replicate inside the cells was also affected. In this system the parasite survival index (PSI) was similar for both tested drugs. L major showed the same response as other tested strains. Geographic information system (GIS) was applied in this study and the ArcGIS software was used to draw a map of the Sudan with different layers and themes. The maps included all the obtained information with coordinates of the studied area. In conclusion: the use of clinical interview combined with simple immunological tests can give valuable information about the pattern of L. donovani infection and predict future prevalence of VL in a short time. Leishmanin non-reactive individuals are a useful piece of data to plan for future vaccine efficacy studies. It is also clear that in vivo drug XII unresponsiveness does not correlate well with in vitro sensitivity for the same drugs. The interactive dynamic map that was produced in the ArcGIS can act as a nucleus for development of a Leishmania network in Sudan and the nearby countries that have the same belt of visceral leishmaniasis.