Mahmoud Romeih

Theodor Bilharz Research Institute

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Mohamed Saber

University of Missouri–Kansas City

Heba Khalil

Theodor Bilharz Research Institute

Tarek S. Abou Shousha

Theodor Bilharz Research Institute

Zhao‐You Tang

Zhongshan Hospital

Mohamed Badawy

Theodor Bilharz Research Institute

Ahmad El-Ray

Theodor Bilharz Research Institute

Hanem Hassan

Theodor Bilharz Research Institute

Xin‐Da Zhou

Ningbo Center for Disease Control and Prevention

Hanem M. Ahmed

Jahangirnagar University

Amr Elkholy

University of Alabama at Birmingham

Cooperative Institutions

Theodor Bilharz Research Institute

Cairo University

Universidade Federal de Minas Gerais

Michigan State University

Children Cancer Hospital

National Water Research Center

National Research Centre

Natural History Museum

Al-Azhar University

Cancer Institute (WIA)

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Advances in the Fight Against Johne's Disease

Sandra Sommer Mahmoud Romeih Paul M. Coussens

More than a century ago, Johne and Frothingham dis- covered Mycobacterium avium subspecies paratuberculosis (MAP) in cattle, the causative agent of Johnes Disease (Paratuberculosis), a chronic, debilitating intestinal infection. Johnes Disease is found most often among domestic and wild ruminants but also has been reported in non-ruminants (1, 15). Whats more, the suggestion that MAP may play a role in human Crohns Disease has led to increased awareness in the medical community and the public (2, 13). Johnes Disease is a frustrating disease for both livestock producers and veterinarians and is associated with large economic losses in dairy cattle (14, 17). Losses are primarily due to decreased milk production and reproduction (5), and reduced salvage value of clinically affected animals. A national study of US dairies found that approx 22 % of farms have at least 10 % of the herd infected with MAP (12). In Michigan, recent estimates suggest that 54% of herds are test positive for Johne's disase (8). The average loss ranges from $40 per cow (low clinical cull rate) to $227 per cow (high clinical cull rate). In this report, we focus on new developments in diagno- sis, vaccines, and control measures in the fight against Johne's Disease. Through on-going national and international research programs, our ability to detect infected cattle is constantly improving and new vaccines are ready for testing. Signs and Transmission

Disease Control

Cattle Diseases

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The frequency of epidermal growth factor receptor mutations in non-small-cell lung cancer patients from Egypt

Egyptian Journal of Pathology (2015)

Noha A.H. Helmy Heba Khalil Shady E.A. Ishak Mahmoud Romeih

10.1097/01.xej.0000462731.13773.18ss

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Characterization Of The Gene Encoding 28 S Large Subunit Ribosomal RNA Of The Lung Stage Of Schistosoma Mansoni (7-Days Schistosomula)

Mohamad Ali Saber Mahmoud Romeih Samir S. Mahgoub

The parasitic helminth Schistosoma mansoni is a major public health concern in many developing countries. Over 200 million people have, and another 600 million are at risk of contracting, schistosomiasis, one of the major neglected tropical diseases. For this dangerous disease the development of long - lasting immunity through vaccination may be the real solution to control the spread of the disease. As molecules on the surface or associated with the tegument of Schistosoma mansoni are a major focus as potential vaccine candidates, but in the present study we screened surface and internal proteins to increase the chances for the discovery of a unique protein of the parasite to be targeted by the immune system of the host and used as a vaccine candidate. In the recurrent study pooled sera collected from Schistosoma mansoni chronically infected patients was purified over a column made of soluble extract of lung stage ( 7-days schistosomula ) of Schistosoma mansoni , then , used to immunoscreen library of 7-days schistosomula, a number of cDNA clones were identified after three rounds of immuno-screening and plaques purification. The phage DNAs of the isolated clones were amplified with polymerase chain reaction (PCR) using forward and reverse primers, then, cloned in PCR TM II plasmid vector. The isolated clone 4-65 was fully sequenced and found to encode the gene of 28S ribosomal RNA of 7-days schistosomula of Schistosoma mansoni. The 0.9 kb cDNA clone was found to have a single open reading frame (ORF) encoding 269 amino acids exhibited 97% identity to the gene of 28S large subunit ribosomal RNA of Schistosoma mansoni and a number of eukaryotic species. (The Journal of American Science. 2008;4(4):77-85). (ISSN: 1545-1003).

Schistosoma

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Analysis of the gene expression profile of Schistosoma mansoni cercariae using the expressed sequence tag approach

Molecular and Biochemical Parasitology (1999)

Túlio M. Santos David A. Johnston Vasco Azevedo Ian L. Ridgers Mercedes F. Martinez

10.1016/s0166-6851(99)00100-0

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Isolation and characterization of Schistosoma mansoni gene coding for antigenic protein from UV irradiated cercariae cDNA expression Library

Mahmoud Romeih Hanem Hassan Mohamed Saber

The attenuated cercariae model is such a good one for facilitating the identification of antigens capable of evoking the protective host immune response which may be the first step towards the production of an effective anti-schistosomal vaccine. Although it is well known that irradiated cercariae is an effective vaccine against schistosomes in the experimental model, the actual effect of irradiation on DNA, RNA and protein to produce this protection is unknown. In this work in the schistosomiasis research we have try to isolate post-irradiation gene transcripts encoding antigens with immunogenic potentiality. SM21.7 was considered to be such a larval stage antigen and its localization in the tegument and subtegument layers confirmed its importance to be used as a target vaccine candidate in case of hosts challenged with S. mansoni. The present study was to clone some gene(s) encoding antigen(s) isolated from UV irradiated S. mansoni expression library that might be vaccine candidates. The antigens were recognized by protective antibodies (IgG fraction) separated from rabbits vaccinated with irradiated cercariae. We have screened UV irradiated S. mansoni cercariae cDNA expression library constructed in lammda ZAPlI, using IgG fraction taken from rabbits vaccinated with irradiated cercariae. The immunoscreening resulted in eight clones that may code for antigenic proteins. Two highly positive clones were isolated and designated UV-Irradiated S. mansoni cercariae 1 and 2 (UVIRSmC1) and UV-IRSmC2. The (UVIRSmC2) clone was determined by restriction enzymes digestion and sequence analysis. The gene has a total size 1088 bp and showed a high degree of identity with differences at 5' end the amino acid level with previously reported S. mansoni antigens: SM21.7 and SMS01. The gene encodes a 21.7 kDa antigen with the highest point of hydrophilicity from amino acid 33 to amino acid 38. The significance of the changes that occurred at the noncoding region sequences of the gene compared to the sequences of SM21.7 and SMS01may have a role in the deference of response to normal and UV irradiated cercariae. (The Journal Of American Science. 2007;3(4):99-112). (ISSN: 1545-1003).

Immunoscreening

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Transforming growth factor β2 is negatively regulated by endogenous retinoic acid during early heart morphogenesis

Development Growth & Differentiation (2010)

Satish Ghatpande Hui‐Ren Zhou Inese Čakstiņa Christopher Carlson Elizabeth A. Rondini

Vitamin A‐deficient (VAD) quail embryos lack the vitamin A‐active form, retinoic acid (RA) and are characterized by a phenotype that includes a grossly abnormal cardiovascular system that can be rescued by RA. Here we report that the transforming growth factor, TGFβ2 is involved in RA‐regulated cardiovascular development. In VAD embryos TGFβ2 mRNA and protein expression are greatly elevated. The expression of TGFβ receptor II is also elevated in VAD embryos but is normalized by treatment with TGFβ2‐specific antisense oligonucleotides (AS). Administration of this AS or an antibody specific for TGFβ2 to VAD embryos normalizes posterior heart development and vascularization, while the administration of exogenous active TGFβ2 protein to normal quail embryos mimics the excessive TGFβ2 status of VAD embryos and induces VAD cardiovascular phenotype. In VAD embryos pSmad2/3 and pErk1 are not activated, while pErk2 and pcRaf are elevated and pSmad1/5/8 is diminished. We conclude that in the early avian embryo TGFβ2 has a major role in the retinoic acid‐regulated posterior heart morphogenesis for which it does not use Smad2/3 pathways, but may use other signaling pathways. Importantly, we conclude that retinoic acid is a critical negative physiological regulator of the magnitude of TGFβ2 signals during vertebrate heart formation.

Heart development

10.1111/j.1440-169x.2010.01183.x

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DNA Immunization with the Gene Encoding SM21.7 Protein Protects Mice Against Schistosoma mansoni Infections

Hanem M. Ahmed Mahmoud Romeih Tarek S. Abou Shousha Mohamed Saber

The current focus of schistosomiasis research is to develop a vaccine that will significantly reduce the incidence of disease. Immunization with DNA is a new trend in vaccine development that could enhance the safety and efficacy of currently used vaccines. The immunogenicity and protective efficacy of a DNA vaccine encoding the antigen SM21.7 was evaluated in C57 BL/6 and Swiss albino mice. The ORF of SM21.7 has been cloned into the eukaryotic expression vector pcDNA 1/Amp under control of CMV late enhancer promoter. The groups of mice were vaccinated intramuscularly with SM21.7-pcDNA1 and boosted twice shown high and specific humoral response in comparison with control (blank pcDNA1/Amp). The level of anti SM21.7 antibody in immunized mice at weeks 3, 6 and 9 intervals post- immunization was significantly higher than in control group. The maximum level of antibodies was obtained at 7 weeks post - challenge infection. Sera from immunized mice could recognize the SM21.7 protein and the specific antibodies were able to mediate a significant killing of schistosomula using peritoneal macrophages as effector cells. In contrast the preimmune sera and sera control serum had no specific reactivity to SM21.7 protein. Immunization with SM21.7- pcDNA conferred a significant level of protection against challenge (41,53%) in Swiss Albino and C57BL/6 mice respectively. Histopathological examination of the vaccinated liver revealed a decreased in the number, size and change in the cellularity of the granuloma compared to the control infected liver. In addition reductions in worm viability, worm fecundity and egg hatching ability have been observed following challenge with S. mansoni cercariae. The number of eggs in the liver and intestine was reduced by 62% and 67% respectively compared to control group. The results suggested that SM21.7 might be a candidate antigen for the generation of antipathology vaccine against schistosomes. (The Journal of American Science. 2006;2(4):59-69).

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PB2000 OUTCOME OF PEDIATRIC NODULAR LYMPHOCYTE PREDOMINANT HODGKIN LYMPHOMA, IS ABVD THE BEST REGIMEN? 10 YEARS SINGLE CENTER EXPERIENCE, REPORT FROM CHILDREN CANCER HOSPITAL, EGYPT.

HemaSphere (2019)

Youssef Madney A. abdallha Sara Ahmed Mahmoud Romeih Sally Fikry

Background: Nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) is an uncommon subtype of Hodgkin lymphoma (HL) in children and accounts for 5–10% of all pediatric HL. Aims: The aim of the study to analyze the clinical characteristics and treatment outcome of Pediatric NLPHL. Methods: Retrospective study including all newly diagnosed patients less than 18 years old with NLPHL and treated in children cancer hospital Egypt CCHE between 2007 and 2017. Results: Among 43 pediatric NLPHL patients, 29 had early stage (18 with stage I, 11 with stage II). Fourteen patients had advanced stage (7 with stage III & 7 with stage IV). Eleven patients had a mediastinal disease, 8 patients had splenic involvement, 2 patient had bulky disease and 10 patient had B symptoms at diagnosis. 28 Patients with Early‐stage disease treatment; surgery and observation 6 patients, Chemotherapy 6 patients and Combined modality 17 patients. Relapse was seen in 7(25%) patients, 1 observation arm, 3 from chemotherapy arm (ABVD) and 3 from the combined arm (ABVD+IFRTH). 14 advanced stage patients received treatment; Chemotherapy (ABVD) 6 patients, Combined (ABVD+IFRTH) 8 patients, Relapse in 7 (50%) patients (all stage 4 disease), 6 from chemotherapy arm and 1 from the combined arm. 2 patients died (stage IV) from progressive disease and sepsis. B symptoms, mediastinal and splenic involvement had a significant impact on DFS on univariate analysis while combined treatment modality (Chemotherapy and Radiotherapy) had an impact on the outcome of both univariate and multivariate analysis. 5 Years DFS, OS for whole group was 64% and 94% respectively, while 5y DFS for early stage 83% and advanced stage 45% (p‐value = .01) and 5 Years OS for early stage was 100% and advanced stage 87 % (p‐value = .08). Summary/Conclusion: Early stage NLPHL had a better outcome but unexpected higher relapse in chemotherapy treatment arm raising a question about the role of radiotherapy while advanced stage disease had poor outcome rising a question of what should be the best chemotherapy regimen for pediatric NLPHL patients.

ABVD

B symptoms

Univariate analysis

Chemotherapy regimen

Dacarbazine

10.1097/01.hs9.0000566492.88815.1b

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Immunization against Egyptian <italic>Schistosoma mansoni</italic> infection by multivalent DNA vaccine

Acta Biochimica et Biophysica Sinica (2008)

Mahmoud Romeih Hanem Hassan Tarek S. Abou Shousha Mohamed Saber

The development of multivalent vaccines consisting of several antigens is a novel approach to creating broad-range protection against different parasite strains and parasite life cycle stages. We have previously confirmed that the schistosome Sm21.7 and SmFimbrin (SmFim) proteins could induce protection in mice. Therefore, this study aimed to construct the multivalent DNA vaccine Sm21.7-SmFim/pBudCE4.1 and evaluate its immune efficacy. The open reading frames of two Schistosoma mansoni genes, Sm21.7 and SmFim, were inserted into the eukaryotic expression plasmid pBudCE4.1 designed for the independent expression of two genes in mammalian cells. To evaluate the in vitro expression of the multivalent Sm21.7-SmFim/pBudCE4.1 DNA vaccine and its immunological effect in mice, the recombinant plasmid Sm21.7-SmFim/pBudCE4.1 was used to transfect 293T cells, and the expression of mRNA and proteins was examined using reverse transcription-polymerase chain reaction and Western blot analysis. Then the ability of Sm21.7-SmFim/pBudCE4.1 to protect against S. mansoni challenge infections was analyzed according to worm burden and egg reduction rates after vaccination of mice. Vaccinated mice showed a significant level of protection (56%), and a decrease in the number and size, and change in the cellular profile, of granulomas. Egg reduction in liver and intestine was 41.53% and 55.63%, respectively, as determined relative to mice that received the empty vector only. In addition to reductions in worm viability, worm fecundity and egg hatching ability were observed following challenge infection in the immunized group. Results showed that Sm21.7-SmFim/pBudCE4.1 could express Sm21.7 and SmFim mRNA and proteins. Enzyme-linked immunosorbent assay and Western blot analysis indicated that immunized mice generated specific immunoglobulin G against Sm21.7-SmFim/pBudCE4.1. These results suggest that vaccination with multivalent S. mansoni DNA vaccine (SmFim-Sm21.7/pBudCE4.1) not only induces a significant reduction in worm and egg burdens, but also significantly reduces the size of egg granulomas. In summary, the multivalent vaccine stimulated specific immunity with a significant level of protection and has anti-pathological effect.

10.1111/j.1745-7270.2008.00404.x

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Applications of polymers in medical technology

Journal of Pharmaceutical Care & Health Systems (2017)

Mahmoud Romeih

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