Role of B cells in the generation of thymus-derived regulatory T cells

Pochodzące z grasicy, naturalne limfocyty T regulatorowe (tTregs, thymusderived regulatory T cells, lub nTregs, natural regulatory T cells), charakteryzujące się fenotypem CD4+CD25+Foxp3+, mają istotne znaczenie dla działania układu odpornościowego ze względu na ich zdolność do hamowania fizjologicznej i patologicznej odpowiedzi immunologicznej oraz kontrolowania odpowiedzi na antygeny własne. Z tego powodu Tregs pochodzące z grasicy odgrywają kluczową rolę w utrzymaniu tolerancji na antygeny własne i homeostazy układu odpornościowego. Limfocyty te powstają w grasicy, po czym migrują do obwodowych narządów limfoidalnych, gdzie podejmują swoje funkcje. Ich rozwój i aktywacja zależą od interakcji z komórkami prezentującymi antygen (APCs, antigenpresenting cells), głównie komórkami dendrytycznymi (DCs, dendritic cells). Rolę APC mogą pełnić również limfocyty B ze względu na powierzchniową ekspresję MHC II i zdolność do pobierania, przetwarzania i prezentacji antygenów limfocytom T CD4+. Celem badań przedstawionych w tej dysertacji była ocena zdolności mysich limfocytów B użytych w roli komórek prezentujących antygen do indukowania rozwoju Treg pochodzenia grasiczego w modelu in vitro, w którym wykorzystano opracowany na podstawie wcześniejszych badań model hodowli całej populacji tymocytów, zawierającej również komórki prekursorowe Treg, z limfocytami B izolowanymi ze śledziony. Znając rolę, jaką pełnią komórki dendrytyczne w różnicowaniu regulatorowych limfocytów T o fenotypie CD4+CD25+Foxp3+ w obwodowych narządach limfoidalnych i w grasicy, zbadano rolę MHC II i cząsteczek kostymulatorowych (CD80, CD86, CD40) aktywowanych limfocytów B na generowanie Treg pochodzenia grasiczego. W celu aktywacji limfocytów B przez receptory TLR7 i TLR4 zastosowano imikwimod (IMQ, Imiquimod) i lipopolisacharyd (LPS). W grasicy naturalnie występuje niewielka populacja limfocytów B, której rola w rozwoju tymocytów, a szczególnie limfocytów T regulatorowych jest mało znana. W niniejszej dysertacji przedstawiono wstępną analizę roli grasiczych limfocytów B w rozwoju nTreg. Ponadto, zbadano rolę glukokortykoidów uznawanych za czynnik selekcyjny w powstawaniu limfocytów T CD4+CD25+Foxp3+ w warunkach hodowli zastosowanych w badaniach. Powszechnie uważa się, że glukokortykoidy, charakteryzujące się silnym działaniem immunosupresyjnym, hamują dojrzewanie komórek dendrytycznych, prezentujących antygen zmniejszając ekspresję MHC II i cząsteczek kostymulatorowych, co w konsekwencji prowadzi do stanu, w którym komórki te częściej indukują tolerancję na antygeny. W badaniach zastosowano syntetyczny glukokortykoid, deksametazon (Dex, dexamethasone). Wyniki badań prezentowanych w dysertacji podzielone zostały na trzy części: pierwsza część dotyczy roli śledzionowych limfocytów B w rozwoju Treg pochodzących z grasicy, w drugiej przedstawiono wstępne wyniki dotyczące roli grasiczych limfocytów B w generowaniu nTreg, a w trzeciej opisano wpływ Dex na powstawanie nTreg w hodowli tymocytów z limfocytami B śledziony. Wyniki badań prezentowanych w pierwszej części wskazują na zwiększenie odsetka limfocytów B śledziony wykazujących ekspresję MHC II i cząsteczek kostymulatorowych (CD80, CD86, CD40) pod wpływem aktywacji. Jednocześnie zwiększeniu ulega ekspresja większości tych białek. W hodowli tymocytów z limfocytami B obserwowano zmianę rozkładu głównych populacji tymocytów, dotyczącą przede wszystkim tymocytów DN CD4–CD8– i DP CD4+CD8+ z małymi różnicami w odsetkach SP CD4+CD8– i CD8+CD4– niezależnie od stosunku tymocytów do limfocytów B i aktywacji limfocytów B. W środowisku hodowli tymocytów i limfocytów B śledziony wykazano zwiększenie odsetka tymocytów SP CD4+Foxp3+. Zjawisko to zależne było od aktywacji limfocytów B i stosunku tymocytów do limfocytów B w hodowli. Wykazano, że limfocyty B śledziony mają zdolność to generowania regulatorowych limfocytów T pochodzenia grasiczego in vitro zależnie od siły sygnałów kostymulatorowych przekazywanych różnicującym się tymocytom. Ponadto, w środowisku aktywowanych limfocytów B śledziony obserwowano zwiększenie ekspresji czynnika transkrypcyjnego Foxp3 w powstałych limfocytach T regulatorowych. Wykazano, że izolowane z hodowli in vitro tymocyty o fenotypie CD4+CD25+Foxp3+ wykazują aktywność supresorową w teście hamowania proliferacji aktywowanych docelowych limfocytów T CD4+. Blokowanie cząsteczek kostymulatorowych i MHC II na limfocytach B wykazało, że CD80/CD86 są skuteczniejsze niż CD40 w procesie generowania nTreg przez limfocyty B, co wskazuje na większe zaangażowanie w ten proces interakcji cząsteczek CD80/CD86 z CD28 niż CD40/CD40L. Rola grasiczych limfocytów B w powstawaniu nTreg in vitro została zaprezentowana w drugiej części rozprawy. Wyniki 72-godzinnej hodowli wskazują, że aktywowane grasicze limfocyty B utrzymują populację nTreg niezależnie od zastosowanego aktywatora (LPS lub IMQ) i przeciwdziałają zmniejszeniu odsetka nTreg, który obserwowano w hodowli niestymulowanej aktywatorami. Wyniki badań pokazują, że grasicze i śledzionowe limfocyty B różnią się od siebie pod względem właściwości po aktywacji i ich roli w indukowaniu rozwoju Treg. Dodatkowo, aktywacja limfocytów B występujących naturalnie w grasicy przez LPS lub IMQ powoduje 2-3-krotne zwiększenie liczby grasiczych limfocytów B po 72 godzinach hodowli w porównaniu z hodowlą 24-godzinną. Te obserwacje pozwalają nam wnioskować, że grasicze limfocyty B wpływają na utrzymanie żywotności lub powstawanie nTreg w sposób zależny od aktywacji limfocytów B i ich proporcji w stosunku do tymocytów. Dodatkowo, interakcja aktywowanych limfocytów B grasicy z tymocytami skutkowała zwiększeniem ekspresji Foxp3 tymocytów o fenotypie SP CD4+Foxp3+. Celem badań prezentowanych w trzeciej części dysertacji było zbadanie zdolności deksametazonu do zwiększenia tolerogenności limfocytów B i w rezultacie do wzrostu odsetka powstających w hodowli nTreg. Przedstawione wyniki wskazują, że w warunkach hodowli limfocytów B śledziony i tymocytów deksametazon nie wykazuje aktywności czynnika selekcyjnego w powstawaniu nTreg.The importance of natural regulatory T cells (nTregs) characterized by CD4+CD25+Foxp3+ phenotype lies in their ability to suppress pathological and physiological immune response, and control the responsiveness to self-antigens. Thus, nTregs have a pivotal role in the maintenance of self-tolerance and immune homeostasis. They are developed in the thymus and migrate to peripheral lymphoid organs to exert their suppressive function. Their development and activation is dependent on the interaction with antigen-presenting cells (APCs), mainly dendritic cells. B cells can also play a role of APC because of their high constitutive expression of MHC II and the ability to uptake, process and present antigens to CD4+ T cells. The aim of the studies presented in this dissertation was to examine the potential of mouse B cells as APC in the generation of thymus-derived nTregs in an in vitro model of co-culture of thymocytes containing nTreg precursors with splenic B cells. Mirroring the role of dendritic cell maturation stage in the differentiation of induced CD4+CD25+Foxp3+ regulatory T cells in lymphoid organs and nTregs in the thymus, we decided to investigate the impact of MHC II and co-stimulatory molecules (CD80, CD86, CD40) expression on B cells upon activation. Imiquimod (IMQ) and lipopolysaccharide (LPS) were used to activate B cells through TLR7 and TLR4, respectively. In addition to (thymic DC and epithelial cell) a small population of B cells can be found in the thymus. Here the role of thymic B cells in the development of nTregs is discussed based on the results of preliminary studies. It is generally accepted that glucocorticoids acting, mainly as immunosuppressive agents, influence dendritic cells maturation by inhibiting the expression of MHC II and co-stimulatory molecules and maintaining the tolerogenic state of antigen-presenting cells. The impact of a synthetic glucocorticoid, dexamethasone (Dex) to induce tolerogenic B cells and to influence the potential of B cells to induce nTreg generation was investigated. The results of the studies presented in this dissertation are divided into three sections; the first section is related to the role of splenic B cells in thymus-derived regulatory T cells development, the second section is focused on the role of thymic B cells in the development of nTregs, and the third section describes the effect of dexamethasone on the generation of nTregs in the co-culture of thymocytes and splenic B cells. The results of the study presented in the first section demonstrated that LPS- and IMQactivated splenic B cells upregulate the expression of MHC II and co-stimulatory molecules that are essential for the presentation of antigens. Both TLR4 and TLR7 activation increased the percentage of B cells positive for particular molecules or their expression facilitating B cells to interact with CD4+ T cells. Co-culture of thymocytes with splenic B cells changed the pattern of the distribution of the main thymocyte subsets mainly DN CD4–CD8– and DP CD4+CD8+ with minor differences in the percentage of SP CD4+CD8– and CD8+CD4– independently on thymocytes : B cells ratio and B cell activation. However, in the microenvironment influenced by the presence of B cells the percentage of SP CD4+Foxp3+ thymocytes was increased. This phenomenon was dependent on the activation of B cells and thymocytes : B cells ratio. Thus, splenic B cells demonstrated the potential to generate nTregs in vitro dependently on the strength of co-stimulatory signals provided to developing thymocytes. In addition, the presence of splenic B cells resulted in the increase of Foxp3 transcription factor expression. The biological activity of nTregs generated in this study was investigated. It was demonstrated that nTregs isolated from cultures maintained under different conditions statistically decreased the percentage of proliferating, activated, responder CD4+ T cells. Blockade of co-stimulatory molecules and MHC II on B cells revealed that the implication of CD80/CD86 and MHC II molecules was more efficient than CD40 molecule in the process of nTreg generation by B cells indicating to the importance of engagement of CD80/CD86 with CD28 molecule, while the engagement of CD40/CD40L is less important in this process. The role of thymic B cells in the generation of nTregs in vitro was presented in the second section. The results of 72 hours of culture revealed that activated thymic B cells maintained the level of nTreg generation independently of the activator used (LPS or IMQ), or counteracted the decrease of these cells observed in non-activated culture. The results revealed that thymic B cells differ from splenic B cells by their characteristics upon activation; in addition, activation by high concentration of LPS (LPS high), or low concentration of imiquimod (IMQ low) induced the increase of thymic B cells about 2-3-fold after 72 hours of culture compared to 24 hours of culture. This observation allows us to conclude that thymic B cells and splenic B cells influence the generation of nTreg in different ways. In addition, the activated cultures showed increased expression of Foxp3 transcription factor. The aim of the studies presented in the third section was to investigate the potential of Dex to render B cells tolerogenic and in consequence to facilitate the generation of nTregs. The results presented in this section showed that Dex induced the decrease of percentage of positive B cells for co-stimulatory molecules as well as for MHC II. Dex did not change the distribution of thymocyte subsets nor did it influence the generation of thymus-derived nTreg cells

Therapeutic design for nasal powder delivery

Coronavirus disease 2019 (COVID-19) is an infectious disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. The response to the pandemic saw widespread diagnostic testing, and the development of vaccines at speed, with global distribution ahead of emergency regulatory approval. Antibody conjugation with nanoparticles (NPs) through Staphylococcus Protein A (SpA) can be used a platform for the development of new diagnostics, and Molecular Dynamics (MD) simulations were used to investigate SpA for its potential as an antibody conjugation agent with model NPs. The simulations showed the model negatively charged silica surface produced favourable SpA adsorption, that facilitated the binding of antibodies at the Fc region to functionalise the system. The SpA was blocked on both sides when it bound to the model gold surface, and the SpA receptor binding domain (RBD) is blocked on the model positive silica surface. Overall, these results indicate SpA is a promising agent to guide the development of a new diagnostic for COVID-19. The initial aim was to develop a novel diagnostic for COVID-19. However, after the withdrawal of my original industrial partner, and the project workplan change with my new industrial partner, it was decided to pursue the development of a dry powder intranasal (IN) vaccine. Therefore, the aim of this thesis is to design a dry-powder IN vaccine for COVID-19, by studying the SARS-CoV-2 RBD interactions with model NPs. The SARS-CoV-2 RBD and SARS-CoV-2 RBD with histidine tag (His-Tag) were studied by MD simulations with model silica NPs (SiNPs) and model carboxyl terminated SiNPs (COOH SiNPs). The MD simulations indicate SARS-CoV-2 RBD binds preferentially on the model SiNPs and SARS-CoV-2 RBD with His-Tag binds preferentially to the model COOH SiNPs. These model systems were used to guide the experimental production of a dry-powder IN vaccine for COVID-19. The simulations guided the laboratory work, where a spray-dryer was used along with other characterisation equipment for the production, and analysis of a dry powder IN vaccine formulation for COVID-19. Alpha lactose monohydrate spheres, used as the excipient in the dry-powder IN vaccine formulation, were successfully spray-dried to the required size. Despite the overall unsuccessful attempt at IN vaccine production for COVID 19, this project has opened up many possibilities for the future, as the MD simulations showed the NPs are suitable for a vaccine formulation, and the experimental component of this project also contributed to the development of an IN vaccine.Coronavirus disease 2019 (COVID-19) is an infectious disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. The response to the pandemic saw widespread diagnostic testing, and the development of vaccines at speed, with global distribution ahead of emergency regulatory approval. Antibody conjugation with nanoparticles (NPs) through Staphylococcus Protein A (SpA) can be used a platform for the development of new diagnostics, and Molecular Dynamics (MD) simulations were used to investigate SpA for its potential as an antibody conjugation agent with model NPs. The simulations showed the model negatively charged silica surface produced favourable SpA adsorption, that facilitated the binding of antibodies at the Fc region to functionalise the system. The SpA was blocked on both sides when it bound to the model gold surface, and the SpA receptor binding domain (RBD) is blocked on the model positive silica surface. Overall, these results indicate SpA is a promising agent to guide the development of a new diagnostic for COVID-19. The initial aim was to develop a novel diagnostic for COVID-19. However, after the withdrawal of my original industrial partner, and the project workplan change with my new industrial partner, it was decided to pursue the development of a dry powder intranasal (IN) vaccine. Therefore, the aim of this thesis is to design a dry-powder IN vaccine for COVID-19, by studying the SARS-CoV-2 RBD interactions with model NPs. The SARS-CoV-2 RBD and SARS-CoV-2 RBD with histidine tag (His-Tag) were studied by MD simulations with model silica NPs (SiNPs) and model carboxyl terminated SiNPs (COOH SiNPs). The MD simulations indicate SARS-CoV-2 RBD binds preferentially on the model SiNPs and SARS-CoV-2 RBD with His-Tag binds preferentially to the model COOH SiNPs. These model systems were used to guide the experimental production of a dry-powder IN vaccine for COVID-19. The simulations guided the laboratory work, where a spray-dryer was used along with other characterisation equipment for the production, and analysis of a dry powder IN vaccine formulation for COVID-19. Alpha lactose monohydrate spheres, used as the excipient in the dry-powder IN vaccine formulation, were successfully spray-dried to the required size. Despite the overall unsuccessful attempt at IN vaccine production for COVID 19, this project has opened up many possibilities for the future, as the MD simulations showed the NPs are suitable for a vaccine formulation, and the experimental component of this project also contributed to the development of an IN vaccine

THE IMPACT OF EXCHANGE RATES ON THE MEASUREMENT AND ACCOUNTING DISCLOSURE IN THE FINANCIAL STATEMENTS IN COMMERCIAL BANKS IN JORDAN

The research aims to clarify the effect of exchange rates on the measurement and accounting disclosure in the financial statements in commercial banks in Jordan and the commitment of the commercial banks listed on the Amman Financial Market in Jordan to disclose the accounting on the translation differences foreign exchange earnings and the statement of the impact of changes in currency exchange rates on the elements of financial statements and also calls for IAS 21: the effects of changes in exchange rates and to identify the relationship between the changes of exchange rates and the elements of financial statements. To achieve the objectives of the study, the researcher used the descriptive method, analytical and interpretative as well as analysis of variance and display graphics, tables, illustrations to clarify the relationships between the variables of the study . The study found that the banks of Jordan is committed to applying the requirements of International Accounting Standard (No. 21): the effects of changes in exchange rates, for each of the translation of transactions denominated in foreign currencies and net investments in foreign subsidiaries and foreign operations which are complementary to the Bank's operations and the translation of vocabulary of the financial statements of foreign affiliates of the Bank, and disclosure requirements of the standard, showing results of the analysis study it. There is also a relationship between each of the earnings per share in commercial banks operating in Jordan and the volume of assets in commercial banks and shareholders equity and cash flows and profits of commercial banks and changes in exchange rates and are affected by. The researcher recommended the need to adhere to the application of IAS 21: the effects of changes in exchange rates and the rest of the criteria because of its importance is reflected on the financial statements and thus the resolution used in the decision-making banks and auditing firms to keep up with new developments in international standards, to take by changes in the the terms of the criteria for modernization, development and activating the role of financial statements in making rational decision, the need to do studies take into account the effects of changes in exchange rates on the financial statements in whole or in part, as well as the need for further studies that take into account the commitment of the Jordanian banks apply other standard

Study of Certain Biomarkers in Iraqi Female Patients with Breast Cancer

صممت الدراسة الحالية لتحديد بعض الماركرات في العراقيات المصابات بسرطان الثدي . تضمنت الدراسة 30 مريضة جمعت عيناتهم النسجية من مستشفى مدينه الطب و بعض المختبرات الاهلية في بغداد بعد موافقة المريضة نفسها و بأستخدام تقنية الكيمياء النسجية المناعية لتحديد هذه الماركرات . اوضحت النتائج علاقة معنوية طردية p ( 0.000 ) بين ماركي النسيج ER و PR و علاقة معنوية طردية p ( 0.000 ) اخرى بين cyclin E phenotype و cyclin E intensity . مما يوضح اهمية هذه الماركرات لسرطان الثدي .The prospective study has been designed to determine some biomarkers in Iraqi female patients withbreast cancer. The current study contained 30 patients whose tissue samples have been collected fromhospitals in Medical City in Baghdad after consent patients themselves and used immunohistochemicaltechnique to determine these markers. The results showed a significant correlation between ER and PR tissuemarkers (Sig = 0.000) and a significant correlation between cyclin E phenotype and cyclin E intensity (Sig =0.001)

Evaluation The MIR26 As a Bio Marker in Rheumatoid Arthritis Patients

Background: Rheumatoid arthritis, also known as RA, is a systemic inflammatory autoimmune disease that is persistent and affects 1% of people globally. Aim of study: This study was conducted to calculation the expression of miRNA-26 by RT-PCR. Methodology: A case control study has been conducted from 15 October 2023 to 15 of January 2024. This study has been done at the Medical City Hospital, Baghdad Government. And included 45 patients ( females and male)  which diagnosed with RA disease by specialist physician, and their ages ranges between, 18 to 75 years. Blood was drawn from the patients to investigate the MIR26, by RT- PCR (Real time polymerase chain reaction). Results: The average Ct value for miR-26 in the group of controls is 6.82, with a fold change of around 1.00 while the average Ct value for miR-26 in group of patients is 7.59, with a fold change of around 7.59. Conclusion: There is significance elevation in miRNA26 in patients group other than in control subjects' group. It is possible that miR-26 contributes to the diseases or disorders that these people suffer from it

Correlation between Serum and Tissue Markers in Breast Cancer Iraqi Patients

            يعد سرطان الثدي من أكثر الأورام الخبيثة انتشارًا بين النساء في جميع أنحاء العالم ، وفي العراق يحتل المرتبة الأولى والسبب الرئيسي لوفيات الإناث المرتبطة بالسرطان. صممت هذه الدراسة لبحث العلاقات بين معلمات المصل والنسيج من أجل توضيح دورها في تطور أو تراجع سرطان الثدي. معلمات الورم هي مجموعة من المواد ، تكون بشكل رئيسي ذات طبيعة بروتينية ، تنتج من الخلايا السرطانية أو من خلايا أخرى في الجسم استجابة للورم. أجريت الدراسة في الفترة من أبريل 2018 إلى أبريل 2019 بإجمالي عدد 60 امرأة مصابة بسرطان الثدي. تم جمع عينات الدم من النساء المصابات بسرطان الثدي في فترة ما بعد الجراحة وما قبل العلاج اللائي حضرن إلى مستشفى الأورام التعليمي في مدينة الطب في بغداد وتم تقييم معلمات المصل بتقنية ELISA وهي Carbohydrate Antigen 15-3 (Ca 15-3) و Carbohydrate Antigen 29 Ca 27.29)) ، هرمون مضاد مولر (AMH) ، عامل نخر الورم ألفا (TNF-α) ، إنترلوكين 6 (IL-6) ، إنترلوكين 10 (IL-10) وبروتين البربخ البشري 4 (HE4) . تم جمع عينات الأنسجة لنفس النساء المصابات بسرطان الثدي اللواتي حضرن إلى مدينة الطب ، بغداد بإجمالي عدد 30. تم تقييم معلمات الأنسجة باستخدام تقنية الكيمياء النسجية المناعية وهي مستقبلات هرمون الاستروجين (ER) ، مستقبلات البروجسترون (PR) ، مستقبل عامل نمو البشرة البشري 2  (Her 2 / neu) و Cyclin E.  أظهرت نتائج العلاقات بين معلمات المصل و معلمات الانسجة وجود ارتباط معنوي موجب (0.017) بين مستضد الكاربوهيدرات 27.29 و مستقبل عامل النمو البشرة البشري 2 ، (0.038) بين انترلوكين-6 مع النمط الظاهري لسايكلين اي ، (0.051) بين عامل نخر الورم-الفا و شدة سايكلين اي ، (0.005) بين بروتين البربخ البشري-4 و مستقبل عامل النمو البشرة البشري 2 و ارتباط معنوي سالب (0.058) بين انترلوكين-10 و مستقبل الاستروجين ، و (0.045) بين بروتين البربخ البشري-4 و شدة سايكلين اي. نستنتج من هذه العلاقات أن العلاقات الطردية تزيد من تطور المرض ، مثل العلاقة بين Ca 27.29 مع Her-2 / neu و cyclin E مع IL-6 و cyclin E مع TNF-α. وقد تساهم العلاقات العكسية في تأخر المرض ، مثل العلاقة بين IL-10 مع ER. من نتائج العلاقات في هذه الدراسة أتضح أن المعلمات Ca 27.29 ، Her-2 / neu ، cyclin E تلعب دورًا مهمًا في تطور المرض.Breast cancer is the most prevalent malignancy among women worldwide, in Iraq it ranks the first among the population and the leading cause of cancer related female mortality. This study is designed to investigate the correlations between serum and tissue markers in order to clarify their role in progression or regression breast cancer. Tumor Markers are groups of substances, mainly proteins, produced from cancer cell or from other cells in the body in response to tumor.  The study was carried out from April 2018 to April 2019 with total number of 60 breast cancer women. The blood samples were collected from breast cancer women in postoperative and pretherapeutic who attended teaching oncology hospital of the medical city in Baghdad and the serum markers evaluated by ELISA technique are Carbohydrate Antigen 15-3 (Ca 15-3), Carbohydrate Antigen 27.29 (Ca 27.29), Anti-Mullerian Hormone (AMH), Tumor Necrosis Factor-Alpha (TNF-α), Interleukin-6 (IL-6), Interleukin-10 (IL-10) and Human Epidiymis Protein-4 (HE4). Tissue samples were collected for the same breast cancer women who attended medical city, Baghdad with total number 30. The tissue markers evaluated by Immunohistochemical technique are Estrogen Receptor (ER), Progesterone Receptor (PR), Human Epidermal Growth Factor Receptor 2 (Her 2/neu) and Cyclin E. The results showed a positive significant correlation (p = 0.017) between Ca 27.29 and Her-2/neu, (p = 0.038) between IL-6 and cyclin E phenotype, (p = 0.051) between TNF-α and Cyclin E intensity, (p = 0.005) between HE4 and Her-2/neu, and negative significant correlation (p = 0.058) between IL-10 and ER score and (p = 0.045) between HE4 with Cyclin E intensity. We conclude from these correlations that positive correlations increasing disease progression, like correlation between Ca 27.29 and Her-2/neu, cyclin E with IL-6 and cyclin E with TNF-α. And the negative correlations may contribute to delay disease, like correlation between IL-10 and ER. From the correlations results in this study, it is clear that the Ca 27.29, Her-2 / neu, cyclin E markers play an important role in disease progression

Detection of Indicator Genes in Methicillin-Resistant Staphylococcus Aureus (MRSA) Isolated from Meat Samples

Aims: The presence of Staphylococcus aureus, particularly methicillin-resistant S. aureus (MRSA), on retail meat products is becoming more and more of a concern. The aim of the present study is to examine the recovered strains of methicillin-resistant Staphylococcus aureus (MRSA) from different retail beef in Najaf/ Iraq. Methodology and results: The phenotypically by conventional identification. Genotypical examination was made also by polymerase chain reaction (PCR) to detect the genes some diagnostic genes for the enterotoxins sea, seb, sed. More than 200 strains of Staphylococcus aureus, from different  butchers meat shops. MRSA strains  were characterized by streaking on different selective media and biochemically identified as Staphylococcus aureus MRSA.  The results showed that the appearance of sea genes in  51% of the isolates. Seb gene was in 18.90% of the isolates while 5.40 %  of the isolates displayed sed gene. Also, Resistance towards six different antimicrobial agents was assessed and revealed that the tested strains of  Staphylococcus aureus showed different level of resistance for  Streptomycin 100%, Cotrimoxazole 100% , Cephalothin  45.90%and Tetracycline 32.3%.  Conclusion, significance and impact of study: the majority of molecular isolate types have been connected to human infections globally, demonstrating that these S. aureus strains in Iraq have a potential for serious pathogenicity. The presence of the genes (sea, sed, and seb) in the MERSA isolates from meat in this study is alarming and raising public health concerns

The Value of Ultrasonography in Diagnosis of Acute Appendicitis in Pediatric Age Group

BACKGROUND Acute appendicitis is one of the most common surgical emergency condition in pediatric age group that need admission to pediatric surgery unit for emergent operation of appendectomy . Any delayed in the diagnosis and operation lead to very serious outcome ,including perforation , abscess formation and appendiceal mass formation and other complications which may lead to high mortality and morbidity rate in case of absence timely performed operation of appendectomy . Ultrasound (USG)is one of the most helpful and informative tool in diagnosis of acute appendicitis in children.it is simple , fast , available and with less complications of ionizing radiation that may associated with other modality of radiological methods , like CT . Aims of the study Evaluate the role of USG examination in :- 1-diagnosis of children with suspected appendicitis 2-defining the sensitivity , specificity and the accuracy rate of acute appendicitis in pediatric age group. 3-Decreasing the normal appendectomy in pediatric age group/. patients and method This prospective study has been achieved in the central child teaching hospital in Baghdad during the period from February 2015 to December 2015, that enrolled 110 patients that were admitted pediatric surgery Center in Central Child Hospital in Baghdad who were complained from right lower abdominal pain and acute appendicitis was highly suspected at time of examination .USG examination had been done to these patients. Result Clinical presentation and USG criteria of acute appendicitis had been found in 57 cases out of 110 patients , and those underwent appendectomy operation in the Central Child Teaching Hospital in Baghdad . Unvisualization of appendix or normal ultrasonography criteria had been found in 53 patients. 6 patients out of the those of 53 patients with negative USG finding underwent appendectomy operation due to persistence of clinical finding, 3 of them were with normal appendix while the 3 others were with inflamed appendix according to histopathological examination. The other 47 patients from 53 cases who were complain from right lower abdominal pain, but with negative USG findings , kept for observation for 24 hours and then discharged home with follow up these cases by clinical examination and USG once weekly for 2 weeks. The overall result of a study is as follow :- Specificity = 100% Sensitivity = 95% Accuracy rate =97.27% Positive predictive value =100% negative predictive value = 50% conclusion ultrasonographic examination have effective role in diagnosis of acute appendicitis in pediatric age group. Keywords:Acute appendicitis , ultrasonogaphy in acute appendicitis , sensitivity , specificity , accuracy rate. DOI: 10.7176/JHMN/73-06 Publication date: April 30th 202

The Role of Saline Enema in Reduction of Intussusception Under Ultra Sonographic Guide a Study of 40 Patients in Alkarama Teaching Hospital

Background: enema reduction of the intussusception under US guide is widely used as a useful alternative to surgical management. This procedure is simple with no radiational effect , less complication rate with good outcome and the parents can stay with their child during the whole procedure. Aim of study : We tried to present our experience in HRIUSG by the use of normal saline enema and to evaluate its safety, outcome and to identify the risk variables for enema failure. Methodology : From the 1st of June 2016 to 1st of January 2017, we received about 45 children at children welfare hospital (medical city complex) that confirmed sonographically to have intussusception. Five patients were excluded from the study because of contraindications (peritonitis, perforation and shock). 40 children were developed enema reduction under sonographic guide. Results : From 40 children underwent HRIUSG their age presentation range from 3 months to 32 month, 67.4 % had respiratory infection while 32.6 % had gastroenteritis. We had a successful rate about 79 % with a median of 1 trial and only 3 patients (9.3 %) developed recurrence within 1st 24 hr. from the 9 patients underwent surgery we had 8 patients had manual reduction and one patient underwent resection of the bowel. Only 3 patients had PLP. 1 patient ( 2.3% ) was reported to have complication ( perforation over sewn ). Age , gender , duration of symptoms not significantly affect the outcome P value < 0.05 while bleeding with stool, initial mass in the left colon and rectum , presence of free fluid and more trial numbers were a bad prognostic factors for enema failure . Conclusion: 1. HRIUSG is simple, safe, less costly, practical, less messy ,no radiational exposure with a low complication rate 2. Age, gender, Duration of symptoms did not affect the outcome significantly. 3. Although bleeding with stool, left colon intussusception mass and free peritoneal fluid is a risk factor for failure of HRIUSG but they are not absolute contraindication. 4. The better scenario for enema reduction success is a patient presented with symptoms < 24 hr. with no bleeding stool and a mass in the cecum and ascending colon with no free peritoneal fluid. Keywords: saline enema; reduction; intussusception ; ultrasound guide . DOI: 10.7176/JHMN/73-04 Publication date: April 30th 202

Flexural Behavior of Reinforced Rubberized Reactive Powder Concrete Beams under Repeated Loads

Non-biodegradability of rubber tires contributes to pollution and fire hazards in the natural environment. In this study, the flexural behavior of the Rubberized Reactive Powder Concrete (RRPC) beams that contained various proportions and sizes of scrap tire rubber was investigated and compared to the flexural behavior of the regular RPC. Fresh properties, hardened properties, load-deflection relation, first crack load, ultimate load, and crack width are studied and analyzed. Mixes were made using micro steel fiber of the straight type, and they had an aspect ratio of 65. Thirteen beams were tested under two loading points (Repeated loading) with small-scale beams (1100 mm, 150 mm, 100 mm) size. The fine aggregate is replaced by 5, 10, and 15%, respectively, with crumb rubber. While replacement of silica fume was 10, 20, 30, and 50%, respectively, with very fine rubber. Also, chip rubber was added to the mixture as coarse aggregate with 5, 10, and 15%. Five tested beams were chosen as case studies to analyze and compare the results of the ABAQUS software with the experimental results. The results showed that the flexural behavior of RRPC beams that contains rubber was acceptable when compared with the flexural behavior of the RPC beam (depending on load-carrying capacity). The crack width was decreased by including waste rubber and steel fibers. There is a satisfactory agreement between the results of the numerical analysis and the results of the experimental testing. Slight ultimate load differences are targeted between the effects of the monotonic loading and the repeated loading.