فهرست مطالب

Archives of Razi Institute
Volume:76 Issue: 4, Sep-Oct 2021

  • تاریخ انتشار: 1400/10/08
  • تعداد عناوین: 45
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  • صفحات 731-739

    بیماری طاعون نشخوارکنندگان کوچک (PPR) یک بیماری به شدت واگیردار است که به عنوان تهدیدی جدی برای صنعت دام کوچک در نظر گرفته می شود. ا این پژوهش با هدف مقایسه ی کینتیک غیرفعال سازی ویروس PPR با استفاده از غیرفعال کننده های مختلف و ارزیابیiPPRV فرموله شده در موش انجام شد. ویروس PPR با استفاده از H2O2 و یا binary ethilenimine (BEI) در دو غلظت 1 و 4 میلی مول غیرفعال شد. سپس ویروس غیرفعال شده با استفاده از ادجوان های مختلف شامل هیدروکسید آلومینیوم (AH)، فسفات آلومینیوم (AP) و یا مخلوط مساوی از AH و AP فرموله شد و در قالب یک طرح کاملا تصادفی با آرایش فاکتوریل 3×3 به صورت درون صفاقی (01/0 میلی لیتر) به 90 سر موش BALB/c (9 سر موش به ازای هر گروه) تزریق شد. واکسیناسیون یادآور 21 روز پس از تزریق اولیه در تمامی حیوانات انجام شد. نتایج نشان داد که استفاده هر سه ترکیب موجب غیرفعال سازی موفقیت آمیز ویروس شد، اما زمان غیرفعال سازی کامل ویروس با استفاده از غلظت 1 و 4 میلی مول BEI و H2O2 به ترتیب 482، 295 و 495 دقیقه پس از آغاز غیرفعال سازی صورت گرفت. اثر اصلی غیرفعال کننده، اندازه گیری شده در روز 42 پس از واکسیناسیون در موش معنی دار بود (05/0>P)، اما اثر ادجوانت و برهمکنش بین غیرفعال کننده و ادجوانت معنی دار نبود (05/0<P). غیرفعال سازی با استفاده از غلظت یک میلی مول BEI نسبت به غلظت 4 میلی مول BEI و H2O2 تیتر آنتی بادی سرمی بالاتری را علیه ویروس PPR در پی داشت (به ترتیب 51/2، 25/2 و 22/2). با این وجود، بین ادجوانت های مختلف از نظر پاسخ آنتی بادی علیه PPR تفاوتی وجود نداشت. در کل، استفاده از غلظت یک میلی مول BEI زمانی که با هر یک از ادجوانت های AH و AP و یا مخلوط آن ها ترکیب شود، موجب افزایش بیشتر پاسخ ایمنی علیه ویروس PPR در موش شد. با این حال، کنتیک غیرفعال سازی مناسب ویروس و همچنین ایمنی زایی مناسبی که با استفاده از H2O2 حاصل شد، در کنار زیست سازگاری بهتر و هزینه ی کمتر، این ترکیب را به کاندیدای مناسبی در آماده سازی واکسن iPPR تبدیل کرده است؛ هر چند باید مطالعات بیشتری در حیوانات هدف انجام شود.

  • صفحات 741-749

    پاستورلوز طیور (وبای مرغی) بیماری مهمی است که پرندگان اهلی و وحشی را در بسیاری از نقاط جهان مبتلا می سازد. این بیماری اغلب توسط سویه های تیپ کپسولی A پاستورلا مولتوسیدا اتفاق می افتد، اگرچه سایر تیپ های کپسولی باکتری نیز گاهی موجب وقوع آن می شوند. مطالعه حاضر با هدف ارزیابی تاثیر برخی آدجوانت ها بر میزان ایمنی زایی و محافظت کنندگی باکترین پاستورلا مولتوسیدا در مرغ و مقایسه آن با واکسن مرسوم تجاری در ایران طراحی گردید. بدین منظور نیمچه های تخم گذار هشت هفته ای دو بار و به فاصله سه هفته مورد واکسیناسیون قرارگرفتند. بررسی ایمنی زایی واکسن ها از طریق آزمون الایزای غیرمستقیم خانگی (In-house indirect ELISA) و با ارزیابی عیار آنتی بادی سرم در روزهای 7، 14 و 21 پس از اولین واکسیناسیون و 14 پس از دومین ایمن سازی انجام شد. عوارض جانبی احتمالی توسط یک متخصص بیماری های طیور ثبت گردید. به منظور ارزیابی میزان محافظت-کنندگی واکسن ها، پرندگان تحت آزمایش دو هفته بعد از دومین ایمن-سازی با LD50 2 (Lethal dose 50) از یک سویه حاد ارگانیسم مورد چالش قرارگرفتند. تعداد جوجه های زنده و سالم تا روز هفت پس از چالش، به عنوان میزان محافظت کنندگی در نظر گرفته شد. نتایج نشان داد که واکسن های حاوی آدجوانت های روغنی مونتاناید ISA 70 و مونتاناید ISA 71 (با یا بدون ساپونین) نسبت به واکسن دارای هیدروکسید آلومینیم و نیز واکسن تجاری موجب پاسخ ایمنی قوی تری شدند (05/0˂P). فقط واکسن های حاوی آدجوانت های روغنی توانستند محافظت قابل توجهی را در مقابل چالش القاء نمایند (05/0˂P). اغلب جوجه های تحت آزمایش التهابات زردرنگی را در محل تلقیح نشان دادند. به نظر می رسد واکسن های ساخته شده با مونتاناید ISA 70 و مونتاناید ISA 71 واکسن های غیرفعال جدید و موثری هستند که قادرند محافظت قابل توجهی را در مقابل بیماری وبای مرغی ایجاد نمایند.

  • صفحات 903-912

    نارنجنین یکی از مهمترین و فراوان ترین فلاونوییدهای شناخته شده است که در گریپ فروت و سایر مرکبات یافت می شود. با توجه به گزارش های مختلف در مورد اثرات ضد التهابی و تعدیل کنندگی سیستم ایمنی بدن از سوی نارینجنین، این مطالعه تجربی با هدف ارزیابی اثرات بالینی و پاسخ های ایمنی آن در مدل حیوانی آرتریت روماتویید (RA) انجام گردید. 40 سر رت نژاد ویستار در محدوده وزنی 180-160 گرم، به طور تصادفی در چهار گروه 10 تایی شامل گروه های: سالم ، کنترل ، درمان شده با نارینجنین و درمان شده با متوترکسات (به صورت خوراکی) تقسیم شدند. برای القای بیماری RA ، ترکیبی از 200 میکرولیتر اجوانت فروند و کلاژن نوع II به صورت زیر جلدی در بالشتک پای عقب رت ها تزریق شد. شدت علایم بالینی RA با استفاده از روش استاندارد امتیاز دهی، ارزیابی گردید. مدت زمان درمان 3 هفته (در خلال روز های 28-7 پس از القاء) در نظر گرفته شد. داده های بدست آمده نشان داد که سطح پروتئین واکنش گر C (CRP)، میلوپراکسیداز ، نیتریک اکسید و سیتوکین های IL-17 و IFN-γ، به طور قابل توجهی در رت های مبتلا به RA افزایش یافت در حالی که سطح آنتی اکسیدان های سرم آنها در مقایسه با گروه سالم به طور قابل توجهی کاهش یافت. میزان التهاب پنجه پا و سطح CRP به طور مشابه در هر دو گروه تحت درمان با متوترکسات و نارینجنین کاهش یافت. در گروه تحت درمان با نارینجنین ، در مقایسه با رت های تحت درمان با متوترکسات، کاهش بیشتری در میلوپراکسیداز، نیتریک اکسید و ظرفیت کل آنتی اکسیدان های سرم مشاهده گردید. با این حال ، سطح سیتوکین های IL-17 و IFN-γ در گروه تحت درمان با متوترکسات، کاهش بیشتری نشان داد. به نظر می رسد استفاده از نارینجنین جهت کاهش اثرات التهابی و کنترل بیماری RA ، مناسب باشد

  • صفحات 935-948

    مطالعه حاضر به منظور بررسی اندوکانابینوییدرژیک و دوپامینرژیک مرکزی در کنترل اخذ غذای القا شده با گرلین در جوجه های گوشتی انجام گرفت. در آزمایش 1، جوجه های 3 ساعت محروم از غذا، تزریقات داخل بطن مغزی (ICV) را به فرم زیر دریافت کردند: سالین، SCH23390 (آنتاگونیست گیرنده 1D دوپامینرژیک، دوز 5 نانومول)، گرلین (6 نانومول) و SCH23390 + گرلین را دریافت کردند. آزمایشات مراحل 2-6 مشابه آزمایش یک بود اما جوجه ها تزریق AMI-193 (آنتاگونیست گیرنده های D2، 5 نانومول)، NGB2904 (آنتاگونیست گیرنده هایD3 ، 4/6 نانومول) و L-741,742 (آنتاگونیست گیرنده های D4 ، 6 نانومول) و L-DOPA (125 نانومول) و 6-OHDA (150 نانومول) را بجای SCH23390 دریافت کردند. در آزمایش 7، سالین، [D-Lys-3]-GHRP-6 (آنتاگونیست گرلین، 5، 10 و 20 نانومول) را دریافت کردند. در آزمایش 8، جوجه ها تزریق سالین، [D-Lys-3]-GHRP-6 (10 نانومول)، دوپامین (40 نانومول) و [D-Lys-3]-GHRP-6 + دوپامین را در یافت کردند. در آزمایش 9، سالین، SR141716A (آنتاگونیست گیرنده های CB1 ، 25/6 میکروگرم)، گرلین (6 نانومول) و SR141716A + گرلین را دریافت کردند. در آزمایش 10، سالین، AM630 (آنتاگونیست گیرنده هایCB2 ، 5 میکروگرم)، گرلین (6 نانومول) وAM630 + گرلین را دریافت کردند. سپس مصرف تجمعی غذا تا 120 دقیقه بعد از تزریق اندازه گیری شد. با توجه به نتایج بدست آمده، تزریق گرلین (6 نانومول) موجب کاهش اخذ غذا شد (P≤0.05). تزریق توام SCH23390 + گرلین موجب مهار کاهش اشتها ناشی از گرلین شد (P≤0.05). تزریق توام L-DOPA + گرلین موجب تقویت اثرات هیپوفاژیک ناشی از گرلین شد (P≤0.05). تزریق توام 6-OHDA + گرلین موجب مهار اثرات هیپوفاژیک ناشی از گرلین شد (P≤0.05). تزریق [D-Lys-3]-GHRP-6 بطور وابسته به دوز موجب افزایش اشتها شد (P≤0.05). درحالی که تزریق توام [D-Lys-3]-GHRP-6 + دوپامین موجب مهار اثرات هیپوفاژیک ناشی از دوپامین شد (P≤0.05). تزریق توام گرلین + SR141716A موجب مهار اثرات هیپوفاژیک ناشی از گرلین شد (P≤0.05). نتایج نشان دهنده این بود که تقابل بین سیستم های اندوکانابینوییدرژیک و دوپامینرژیک و گرلین از طریق گیرنده های D1و CB1 در جوجه های گوشتی میانجی گری می شود.

  • صفحات 1047-1053

    باکتری آوی باکتریوم (هموفیلوس) پاراگالینارم عامل بیماری کریزای عفونی در ماکیان است. علی رغم انتشار جهانی این بیماری در ایران مطالعه کمی در خصوص جداسازی و شناسایی آن صورت گرفته است. این مطالعه بمنظور جداسازی و تعیین حساسیت انتی بیوتیکی جدایه های حاصل از مرغان بومی مشکوک به بیماری در بازار پرنده صورت گرفت. از مجموعه 18 مورد نمونه سواب ناحیه کامی تهیه شده تعداد 4 (22 درصد) جدایه باکتری مشکوک به اوی باکتریوم پاراگالینارم با ویژه گی های رشد اقماری و ازمون های بیوشیمیایی کاتالاز و اکسیداز بدست امد که با آزمایش PCR ویژه گونه اوی باکتریم مورد تایید قرار گرفتند. همچنین ازمون مستقیم PCR بر روی نمونه های سواب 12 مورد (66درصد) مثبت مشاهده شد. آزمون حساسیت آنتی بیوتیکی به روش انتشار دیسک کریر-برویر با استفاده از ژلوز خون اسب محیط کلمبیا انجام شد. مقاومت به آموکسی سیلین، اکسی تتراسکلین، استرپتومایسین، تریمتوپریم/ سولفادیازول (75%) و حساسیت به سفالکسین سفتریاکسون، انروفلوکساسین، فلورفنیکل، جنتامایسین، لینکومایسین، نیومایسین، داکسی ساکلین(50%)، دانوفلوکساسین (75%) فلوموکویین (50%)، افلوکساسین(75%) مشاهده شد. حاله مهار رشد متوسط باکتری در برابر آمپی سیلین، کولستین، اریترومایسین، پنی سیلین، تیامولین (75%) تایلوزین(75%) دیده شد. بطور خلاصه این مطالعه نشان داد شیوع کریزای عفونی در مرغان بومی شایع بوده و ممکن است از نظر بیماری منبع خطری برای گله های صنعتی و نیز مقاومت داروی اوی باکتریوم پاراگالینارم هم برای مرغان و بهداشت عمومی بطور غیر مستقیم باشد. مطالعه عمیق تری برای تهیه واکسن بومی ارزان قیمت ویژه مرغان بومی جهت کاهش بیماری و مقاومت های میکروبی توصیه میگردد.

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  • A. Gasmi *, P. Mujawdiya, S. Noor, S .Piscopo, A. Menzel Pages 707-719

    Obesity becomes a chronic disease due to the increasing number of mortality and morbidity cases around the world. In most regions, chronic illnesses, such as obesity, are important sources of morbidity and mortality. Due to a lack of effective strategies for prevention and management, the adverse effects of obesity and related diseases on health continue to be a serious problem. Relevant information was searched from Google Scholar, Scopus, and PubMed using such different terms as “Obesity”, “Obesity Management”, “Obesity AND Physical activity”, “Obesity AND Genetics”, “Obesity AND Diet”, and “Obesity AND Nutrigenomics”. Obesity is characterized by a complex interaction of hereditary and lifestyle factors, which includes food. Diet is an environmental element that plays an important and considerable role in the management of health and reduces the risk of obesity and its comorbidities. Changes in lifestyle patterns not only help burn extra calories but also prevent the development of obesity via its modulating effect on genetic factors. Different people respond differently to an obesogenic environment. The notion of nutrigenetics emerged as a result of various genetic variations that may explain this heterogeneity. Nutritional genomics, also known as nutrigenetics, is the study that investigates and analyses gene variations linked to varied responses to certain foods; moreover, it links this variation to diseases, such as obesity. As a result, tailored nutrition advice based on a person's genetic profile may improve the outcomes of a specific dietary strategy and offer a novel dietary strategy to improve life quality and preventing obesity. This study concluded that physical activity and dietary interventions play an effective role in the management of obesity. Moreover, understanding of the function of the most prominent obesity-related genes, as well as the interaction between nutrition and gene expression, will help researchers design personalized treatment strategies for humans.

    Keywords: Diet, Obesity, Physical activity, Nutrigenetics, Single Nucleotide Polymorphisms
  • K. Sharun *, K. Jambagi, K .Dhama, R .Kumar, A. M. Pawde Amarpal Pages 721-730

    Platelets are the reservoir of growth factors and play a major role in several physiological processes, such as coagulation, angiogenesis, immune response, and tissue repair. Platelet concentrates are broadly classified into two groups depending on their fibrin content, namely platelet-rich plasma (PRP) and platelet-rich fibrin (PRF). They are further divided based on their leucocyte contents. The PRP is plasma containing supra-physiological concentrations of the platelets. The growth factors present in the PRP play a crucial role in the promotion of local angiogenesis, regulation of cellular activity, stem cell homing, proliferation and differentiation of different stem cells, and deposition of matrix proteins contributing to tissue regeneration. This review aimed to establish the therapeutic potential of PRP in canine medicine with a particular focus on the applications in ophthalmology, dermatology, and musculoskeletal disorders. A systematic literature review was performed to identify the literature published during the past 20 years (2001-2021) using authentic academic databases, such as PubMed, Science Direct, Google Scholar, and Scopus. In the initial search, 556 articles were identified and based on the specific inclusion and exclusion criteria, 59 articles were selected for further analysis. The clinical efficacy of PRP depends on the number of platelets and the growth factor concentration. The PRP-based biological therapy has broad clinical applications in musculoskeletal pathologies. It is a simple, safe, and cost-effective method that can be used to treat various diseases and disorders in canine practice. For example, PRP is used for managing corneal ulcers, corneal erosion, alkali burn, keratoconjunctivitis sicca, burn wounds, chronic wounds, cutaneous ulcers, acute traumatic bone fractures, tendinopathies, cartilage pathologies, osteoarthritis, and abdominal wall defects either as monotherapy or as an adjunctive therapeutic agent. In addition, PRP is widely used as a carrier of mesenchymal stem cells for transplanting into bone defects. Therefore, allogeneic PRP therapy can be considered a simple, safe, and cost-effective method for the treatment of various diseases and disorders in canine practice. The therapeutic application of PRP in canine medicine is limited in the present study due to the lack of consensus for collection, characterization, and clinical use. Hence, further studies are required to establish the actual worth of PRP-based regenerative strategies in canine medicine.

    Keywords: canine medicine, canine practice, dermatology, platelet-rich plasma, regenerative medicine
  • M .Akbarian, H. Keyvanfar *, M. Lotfi, S. M .Azimi Dezfuli, H. R. Varshovi Pages 731-739

    Peste des petits ruminants (PPR) is a highly contagious disease that is considered a major threat to the small livestock industry. Although vaccination via live-attenuated PPR vaccine is a main controlling strategy in the endemic area, during PPR eradication process, the inactivated PPR vaccine (iPPRV) is recommended. This study aimed to compare the inactivation kinetics of the PPR virus via different inactivants and immunogenicity evaluations of the iPPRV formulated vaccine in mice. The vaccinal live PPR virus was inactivated by either H2O2 or binary ethylenimine (BEI (at two concentrations of 1 or 4 mM. Thereafter, the inactivated virus was formulated with different adjuvants, including aluminum hydroxide (AH), aluminum phosphate (AP), and a mixture of AH and AP that were intraperitoneally (IP) administrated (0.1 mL) to 90 BALB/c mice in a completely randomized design and 3×3 factorial arrangement (9 animals per group). The booster vaccination was carried out in all animals 21 days after the primary vaccination. Results showed that the PPR virus was successfully inactivated by all the inactivation agents; however, the time of complete virus inactivation was estimated to be 482, 295, and 495 min post-treatment initiation for 1 mM BEI, 4 mM BEI, and H2O2, respectively. The main effect of inactivant on antibody titers against PPR virus that was measured after 42days post-immunization in mice was significant (P<0.05); however, the adjuvant and interaction effect of inactivator×adjuvant were not effective(P>0.05). Inactivation by 1 mM BEI was associated with a higher antibody titer against PPR virus (P<0.05) in comparison with both 4 mM BEI and H2O2 (2.51 vs. 2.25 and 2.22, respectively). Meanwhile, there were no significant differences among the used adjuvants in terms of eliciting antibody response against PPR virus. In conclusion, the use of 1 mM BEI in combination of AH, AP, or a mixture of AH and AP was associated with a higher immune response against PPR virus in mice. However, the appropriate inactivation kinetic of the virus and immunogenicity associated with the use of H2O2, as well as its biocompatibility property and better cost-benefit, nominated H2O2 to be used in iPPR preparation; however, more investigations are required in target animals.

    Keywords: Immunogenicity, Inactivation, Peste des petits ruminants (PPR), Vaccination, Virus
  • R. Ghadimipour *, M .Ghorbanpoor, D. Gharibi, M. Mayahi, A. R. Jabbari Pages 741-749

    Avian pasteurellosis (fowl cholera) is an important disease affecting domestic and wild birds all over the world. Although the capsular type A of Pasteurella multocida is mostly involved, other capsular types are occasionally incriminated. The present study aimed at investigating the effect of some adjuvants on immunogenicity and protectivity of P. multocida bacterin in chickens, compared to an Iranian commercial vaccine. Eight-week-old chicken pullets were double vaccinated with an interval of three weeks. Vaccine immunogenicity testing was conducted using an in-house indirect enzyme-linked immunosorbent assay and assessing serum antibody titers at 7, 14, and 21 days post-primary and 14 days post-secondary immunization. The possible adverse effects were recorded by a poultry-disease expert. For evaluating the vaccine protection rate, chickens were subjected to 2×Lethal Dose 50%of a virulent P. multocida strain two weeks post-secondary immunization. The rate of live and normal animals was regarded as protection rate 7days after the exposure. The findings showed that oil adjuvants Montanide ISA 70-and Montanide ISA 71-containingvaccines (with or without saponin) caused a powerful immune reaction than the aluminum adjuvanted vaccine and commercial vaccine (P<0.05). Significant protection against challenge was merely induced by the oil adjuvanted vaccines (P<0.05). The majority of the studied chickens showed inflammation at the injection site (yellow) throughout the trial. Vaccines made by Montanide ISA 70 and Montanide ISA 71 are novel and effective inactivated vaccines that are able to cause significant protection to fowl cholera disease.

    Keywords: Adjuvants, Chicken, Pasteurella multocida, vaccine
  • K. Ahmad Khidir * Pages 751-759

    It has been a few months since the first batch of Coronavirus Disease 2019 (COVID-19) vaccines arrived in the Kurdistan region, and the priority was given to health workers at the forefront of the treatment of COVID-19 patients. The rollout is slow, and there is little evidence to suggest that the whole Kurdistan region is vaccinated anytime soon. This comprehensive and national survey was conducted to investigate the perception of the people of the Kurdistan region towards COVID-19 vaccine hesitancy. An adjusted valid and dependable questionnaire was deployed via social media platforms (Facebook and Viber) to invite participants aged 18 and over from the residents of the four provinces of the Kurdistan region. A total of 450 individuals participated in this study. The majority of the participants were male (54.4%) who were aged 26-40 years with bachelor's degrees (44.3%). Moreover, they were full-time employees (37.8%) with a household income of 0-$5,000 (53.3%). They were the residence of urban regions (81.9%) and Sulaymaniyah province (87.7%). On the probability of getting a COVID-19 vaccine shot, the responses were very likely (26.7%), somewhat likely (24.9%), not likely (20%), and definitely not (28.4%). The vaccine hesitancy prevalence was high among individuals aged 26-40, students with low incomes, unemployed, and those from the suburban areas, while respondents with the least perceived threat to get infected with the COVID-19 in the next year had the highest level of vaccine hesitancy. It is evident that vaccine hesitancy is high, and multiple strategies across the Kurdistan region needed to be implemented to encourage people to get vaccinated; therefore, scientific communication is necessary with the help of mass media.

    Keywords: COVID-19 vaccine, Vaccine hesitancy, Kurdistan region, Nationwide assessment
  • A .Saud Hussein *, N .Ibraheem Salih, I .Hashim Saadoon Pages 761-768

    Breast cancer is the most frequent cancer among women and causes the greatest number of cancer-related death among women all over the world. It approximately accounts for 15% of all cancer death. The human microbiota is the term applied to the aggregate of microbes that live in different habitats of living organisms 'bodies, including the gut, skin, vagina, and mouth,  as well as nose, conjunctiva, pharynx, and urethra, among others. Increasing evidence is pointing to the role of the microbiome in the occurrence and development of a variety of cancers. Intestinal microbiome imbalance is related to the occurrence of gastrointestinal tumors, such as esophageal, gastric, colorectal, and gallbladder cancer. The present study aimed to identify the role of microbiota in the development of breast cancer. The women with breast cancer (n=130) in this study were in the age range of 25-75 years. The study was conducted in Kirkuk city of Iraq from September 10, 2019, to March 15, 2020. The control group included 20 women diagnosed with benign breast lesions in the age range 25-75 years, who matched the women in the patient group. Blood samples and breast tissue samples were taken from patients with breast cancer and benign breast lesions. Blood samples were examined through immunological methods, enzyme-linked immunosorbent assay (ELISA) was adopted for the detection of interleukin-19 (IL-19). Breast tissue samples were taken from breast cancer and benign breast lesions patients to isolate and identify bacteria. Based on the obtained results, only 6 out of 30 (20%) cultured breast tissue samples from women with breast cancer showed bacterial growth.  In total, 4 (67%) and 2(33%) of these 6 positive cultures were Escherichia coli was and Staphylococcus aureus, respectively, and this relation was statistically significant. However, no bacterial growth was observed on the cultured breast tissue samples taken from women with benign breast lesions. Moreover, the difference between women with a positive and negative result of bacterial culture and stages of breast cancer was statistically non-significant.  It is worth mentioning that 50 % of women with breast cancer and bacterial growth were within the age range of 40-49 year. The present study revealed that the difference between women with breast cancer and those with benign breast lesions was statistically highly significant according to the place of residence. In addition, the mean level of IL-19 among women with breast cancer was lower than that in women with benign breast lesions, and this relation was statistically highly significant.

    Keywords: Breast cancer, IL-19, Microbiota
  • D .Hazim Abdul Hameed *, E .Hussein Ali Pages 769-779

    The bacterial isolates Streptomyces were obtained from the soil and cultivated in a wheat bran medium, which was used to produce the L–glutamate oxidase enzyme. The extracellular enzyme was then extracted using a cooling centrifugation process to obtain the filtrate that represents the crude enzyme. Afterward, the enzyme purification processes were carried out which included precipitation with ammonium sulfate as a preliminary purification step followed by dialysis to remove the salts. Next, ion-exchange chromatography and gel filtration were used to finish the purification process, and the enzyme activity was determined for each purification step. The results of purification of L-glutamate oxidase enzyme from streptomyces using ammonium sulfate showed that the specific activity was 8.25 units/mg protein with a saturation ratio of 60%. Moreover, the results of purification using a dialysis tube indicated that the specific activity was 9.5 units/mg protein. In addition, the result of purification using diethylaminoethyl cellulose ion column revealed that the specific activity was 25 unit/mg protein and the results of purification using gel filtration showed that the specific activity was 56 units/mg protein which was the best step in the purification process due to high specific activity of the enzyme. The optimum temperature and pH for the activity and stability of the enzyme were tested. Based on the findings, the optimum temperature for the activity of the enzyme was 37 °C. In addition, it was found that the optimum temperature range for the stability of the enzyme was 30-50 °C. Besides, the optimum pH for the activity was 7.0 and the optimum pH range for the enzyme stability was 5.0-7.0.

    Keywords: L-glutamate oxidase, L-glutamate, Streptomyces, Specific activity
  • R .Abduladheem Jabbar *, N .Neima Hussien Pages 781-793

    The biosynthesis of silver nanoparticles (AgNPs) is a new approach in nanotechnology which was optimistically implemented in medicine, food control, and pharmacology. The present study aimed to investigate the antioxidant and cytotoxicity effects of AgNPs produced by Lactobacillus gasseri filtrate. Also, changing color from yellow to brown confirmed the production of AgNPs. AgNPs were characterized using ultraviolet-visible spectroscopy, FE-SEM, and Fourier Transform Infrared Spectroscopy (FTIR). The antioxidant activity of AgNPs was tested using the DPPH assay. The scavenging test for DPPH showed 19.3%, 32.6%, 47.6%, 72%, 85.3% at concentrations (6.25, 12.5, 25, 52, 100) µg/ml, respectively, which proved that the scavenging percentage increased with increasing concentration. The effect of AgNPs on the chromosomal pattern was also studied. The results of the experiment of AgNPs against SK-GT-4 cancer cells showed the toxic activity of the used particles against the strains of these human esophageal cancer cells and failed to affect normal cells.

    Keywords: cancer cells, Chromosomal Aberrations, Nanoparticles
  • M. Al Rawi, N. H. A. L. AlMudallal *, A. A .Taha Pages 795-808

    The improvement of multi-resistance properties of the bacterial pathogen has recently been discussed as an emerging issue. In this regard, iron oxide nanoparticles have attracted the researchers’ attention due to their wide application in the realm of medicine. Iron oxide nanoparticles have a high specific surface area that enables them to interact with the bacterial surface structure and has considerable antibacterial activity. The current study aimed to synthesize a novel antimicrobial agent from iron oxide nanoparticles and determine its minimum inhibitory concentration (MIC) on different gram-positive and negative variant bacterial strains isolated and characterized from the infected urinary tract of Iraqi elderly patients. This study was conducted from September 2020 to December 2020 on 75 urine samples collected from the infected urinary tract of elderly patients in the ages range of 60-75 years admitted to Al-Yarmouk Medical Hospital, Baghdad, Iraq. Isolation of bacterial isolates was carried out using differential and selective media. Afterward, they were characterized and confirmed using different biochemical tests and VITEK 2 system, respectively. Magnetic nanoparticles were fabricated by co-precipitation of ferric ions (Fe3+) and ferrous ions (Fe2+) in presence of ammonium hydroxide solution (25%). The characterization of synthesized nanoparticles was performed subsequently using UV-VIS spectroscopy analysis, Scanning Electron Microscope (SEM), Fourier transform infrared spectroscopy analysis, X-ray Diffraction analysis (XRD), and Energy-dispersive X-ray spectrum (EDX). The MIC of synthesized sonicated Fe3O4NP against different bacterial strains was determined using the broth culture dilution method through making serial dilutions of 50, 100, 200, 400, 500, 600, 800, 900 µg/ml from a 5mg/ml nanoparticle stock solution. Afterward, the lowest concentration of nanoparticles required to arrest the growth of bacteria was determined through the colony-forming unit of each treated bacteria on brain heart infusion agar. In total, 17bacterial isolates were identified from the infected urinary tract, five bacterial isolates (E. coli, Pseudomanas aeruginosa, Staphylococcus aureus, Enterococcus faecalis, and Micrococcus luteus). In addition, two Proteus mirabilis strains were identified separately and were tested against synthesized Fe3O4NP to determine the MIC. The novel synthesized antibacterial agent showed excellent bioactivity, compared with controls (consisting of bacterial suspension without ferrous oxide nanoparticles), and the synthesized antibacterial agent was considered significantly active against all the bacterial strains at a p-value less than 0.05. The Fe3O4NP were active against gram-negative more than gram-positive bacteria. The MIC of synthesized and characterized Fe3O4NPwas applied on seven gram-positive and negative bacterial isolates using bacteria- Fe3O4NP complex. Significant effects were observed on all strains, compared with controls, and this complex could significantly inhibit gram-negative more than gram-positive bacteria.

    Keywords: Iron Oxide Nps, Bacterial Strains, Uti Elderly Patients, Minimum Inhibitory Concentration
  • B. H .Jasim *, E. H. Ali Pages 809-820

    Pseudomonas aeruginosa was isolated from injuries of patients' wounds and burns, and to ensure that the isolate was belonging to P. aeruginosa, several tests were performed, such as staining techniques, a biochemical test, morphological test, Vitek 2 system, and sensitivity test. The results of the gram stain test showed rod pink gram-negative bacteria, demonstrating that the isolate belonged to P. aeruginosa. Growth optimization of bacterial was performed by assessing different combinations of pH and temperatures. It is revealed that the best conditions for increasing the number of bacteria were achieved at 37°C with the bacterial number of 5.53×108 and pH 6 with the bacterial number of 5.87×108. Fibrinolytic enzyme is an agent that lysis fibrin clots. This fibrinolytic factor has prospective use to treat cardiovascular diseases, such as stroke and heart attack. Cardiovascular diseases have attracted worldwide attention for their elevation morbidity and mortality. Fibrinolytic enzyme was extracted by centrifugation at 10000 × g at 4°C for 10 min, the supernatant was kept and the pellet having bacterial cells was discarded. Purification of the fibrinolytic enzyme was achieved using salt precipitation, ion exchange, and gel filtration chromatographic techniques. The results showed that the gel filtration chromatography had optimal specific activity and purification fold at 562.6 U/ml, and the final specific activity of the purified enzyme increased 4.1 times. The molecular weight of the fibrinolytic enzyme was determined at26 kDa by gel filtration chromatography. The purified fibrinolytic enzyme had optimum activity atpH 7 and40°C. The pH stability for the enzyme activity was found in pH 6-7 and the range of 10-40°C.

    Keywords: Characterization, Isolation, identification, Pseudomonas aeruginosa, Fibrinolytic enzyme
  • B. Saeed *, S. Al-Jadaan, B. A Abbas Pages 821-827
    New medicinal compounds are being evaluated due to the increasing prevalence of cancer in human societies and the necessity to produce new medications for treatment. The new Schiff base compound 4,4'-[1,4-phenylenebis(1,3,4-thiadiazole-5,2-diyl)] bis (azaneylylidene) bis (methaneylylidene) diphenol, which was previously produced from the reaction of 5,5' [(1,4-Phenelene) bis (1,3,4-thiadiazol-2-amine)] and the para-hydroxy ben aldehyde was synthesized and different concentrations (250 and 300 mg/mL) of this new compound were exposed to breast cancer (MCF-7) cells to examine its cytotoxicity effect. Cell line viability, acridine orange/propidium iodide staining, and DNA fragmentation were assessed in evaluating the antitumor effect of the new composition. Obtained data from cell viability assays demonstrated cytotoxic activity against MCF-7 breast cancer cell lines. No fragmentation was observed in DNA fragmentation of the novel compound base with MCF-7 and Vero cell line. The new Schiff base compound indicated well-defined anti-cancer activity when treated with breast cancer cells (MCF-7). The compound blocked the proliferation of cancer cells without apoptosis. As a consequence of the findings, it was recommended to use this compound in treating breast cancer.
    Keywords: Breast cancer, DNA Laddering Assay, Thiadiazole Compound
  • S. Abdul Raheem Hasan *, S. Sajid Al Jubori, J .Abdul Sattar Salman Pages 829-840

    Specialized Escherichia coli (E. coli) isolates, called uropathogenic E. coli (UPEC), cause most of urinary tract infections (UITs). Once bacteria reached the urinary tract of the host, they have to adhere to the host cell for the colonization. For this purpose, bacteria have different structures including fimbrial adhesins. Most of the UPECs contain type 1 fimbriae encoded by fim operon (fimB, E, A, I, C, D, F, G, H) which is responsible for the adhesive ability in these isolates. Ninety-four isolates of UPEC were obtained from UTI patients in Baghdad hospitals and their diagnosis were confirmed by the PCR method using 16srDNA as a housekeeping gene. The UPEC isolates were tested for their ability of adherence to the urothelial cells obtained from the mid-stream urine from healthy women. Fifty isolates were subjected to detect type1 fimbriae genes (fimA operon) using specific primers followed by sequencing the amplified fragment which they were analyzed by Geneious software. The results confirm that all the isolates were E. coli according to the genetic analysis by the PCR test, and also, the ability of attachment for all isolates were approved (100%).For type 1 fimbriae, the findings figured out that 100% of the isolates harbored fimA,fimI, fimC, fimD, fimG and fimH genes; while 96% of them were positive for fimB, fimF,and 82% of the isolates were positive for fimE. This result exhibited a higher prevalence of fim genes, as the attachment ability was 100%. Approximately, all UPEC have type 1fimbrial genes, so it could be used as a genetic marker in the investigation of E. coli adhesion ability.

    Keywords: fimA operon, Uropathogenic Escherichia coli, attachment ability
  • B .Kareem Queen *, A. Abdullah Al Janabi Pages 841-846

    Several previously published reports have suggested a relationship between type 2 diabetes mellitus (T2DM) and chromosome 10q. The results of genotyping of 228 microsatellite markers in Icelandic people with T2DMrevealed that a microsatellite, DG10S478, within intron 3 of the transcription factor 7-like 2 gene (TCF7L2; formerly TCF4) was associated with T2DM. The present study was aimed to analyze the sequence of TCF7L2 in Iraqi patients with T2DM. This study was performed on the blood samples of 10 patients within the age range of 18-70 years old with T2DM. The DNA was extracted from the whole blood samples and the TCF7L2 gene was purified and amplified using the polymerase chain reaction (PCR) technique. Afterward, the PCR products were run in gel electrophoresis to detect the gene. Moreover, the BLAST software was used to analyze the gene TCF7L2 sequence which was compared with the reference sequence of the template gene from NCBI. The results of TCF7L2 gene sequences obtained from the samples collected from the Iraqi patients with T2DMwere received from Macogen Company, Korea, and analyzed using the BLAST software. The findings showed mutations in the gene sequence of all patients, compared to the gene sequences in NCBI. Hence, the mutation in the TCF7L2 gene was present in Iraqi patients with T2DM, and it could be one of the factors causing and increasing the risk of T2DM disease.

    Keywords: Bioinformatics Methods, Diabetic mellitus type 2, mutation, TCF7L2 gene
  • R. Khandia *, B. Pattnaik, K. Rajukumar, A. K. Pateriya, N .Puranik, S .Bhatia, H. Murugkar Pages 847-855

    Edema factor (EF) is one of the major secretory proteins of anthrax bacteria along with protective antigen (PA) and lethal factor (LF). Edema factor is a calmodulin-and calcium-dependent adenylate cyclase that increases intracellular levels of cAMP. Intracellular trafficking of EF occurs through PA by binding to ATR/CMG2 receptors, which are also involved in other physiological functions of cells. cAMP is a secondary messenger which activates multiple signaling cascades involved in the cytokinetics of actin molecules and cell junction formation. The present study evaluated the effect of EF on growth and angiogenesis patterns in chicken embryos in the in ovo model. Angiogenesis in the chorioallantoic membrane (CAM) of an embryonated chicken egg was decreased and embryo growth was delayed by EF despite the absence of trafficking moiety PA, which is required for transferring the EF molecule inside the cell. Angiogenesis inhibition and embryo growth retardation indicate the use of an alternative receptor by EF to modulate these cellular functions. Additionally, docking was performed between EF as a ligand and hepatocyte growth factor receptor (cMET) and vascular endothelial growth factor (VEGF) receptors, which are mainly involved in growth and angiogenesis. The analysis revealed a very strong binding of EF to cMET receptor (in terms of the number of hydrogen bonds and energy) compared to its ligand hepatocyte growth factor (HGF), which indicates the use of cMET receptor by EF and induction of angiogenesis and embryo growth retardation possibly by competitive inhibition of HGF ligand or receptor-mediated endocytosis.

    Keywords: angiogenesis inhibition, Edema factor, CAM, embryo growth retardation, CAMP, signaling molecule
  • R. A. Hikmet *, N. N .Hussein Pages 857-869

    This study conducted a mycosynthesis of silver nanoparticles (AgNPs) by Candida albicans supernatant. The mycosynthesized AgNPs were identified by color visualization, ultraviolet-visible (UV) spectroscopy device, X-ray diffraction (XRD), energy dispersive analysis of X-ray (EDX), field emission scanning electron microscope (FESEM), and zeta potential analysis. The UV-Vis spectroscopy examination has shown the highest absorbance (λmax) at the wavelength of 429 nanometers, which was the indicator of the creation of AgNPs. Furthermore, XRD showed the crystalline structure of AgNPs, and EDX revealed the weight percentage of silver atoms in the sample (82.4%). According to the FESEM, the morphology of AgNPs was spherical, and its size was 40.19 nanometers. Zeta potential analysis indicated that AgNPs were middling stable in the solution, and the zeta potential of AgNPs mycosynthesized by C. albicans was-23.02 mV. The cytotoxic effect of AgNPs against a human colon cancer cell line using MTT assay has shown the presence of toxic action against the cells, and no cytotoxic effect appears on the normal cells. The antioxidant activity of AgNPs using DPPH assay demonstrated 17.0%, 29.3%, 48.3%, 67.6%, and 83.6% at concentrations of 6.25, 12.5, 25, 50, and 100 µg/ml, respectively. The impact of AgNPs on the chromosomal pattern has also been studied. The importance of this study lies in the possibility of the synthesis of AgNPs using this yeast since most nanoparticle preparation methods utilize molds.

    Keywords: AgNPs, Antioxidant, Candida albicans, Cytotoxicity, EDX, FESEM, Nanoparticles, UV Spectroscopy, XRD, zeta potential
  • I. M. Aufi *, A. M .Khudhair, L. Ghaeb AL Saadi, M. A .Almoneem Ahmed, F. M. Mahdi Shukur Pages 871-877

    The importance of influenza viruses in respiratory infections in the Middle East, including Iraq, has been historically overlooked. Nowadays, with the pandemic of corona virus disease 2019, the importance of prevention from other respiratory diseases, such as seasonal influenza, can be a critical step in the health management system. Therefore, this study aimed to evaluate the prevalence and seasonal occurrence of influenza viruses in the Iraqi population presented with influenza‐like illness (ILI) or severe acute respiratory infection (SARI)within2015-2017. Moreover, this study was conducted to identify the periods with increased influenza transmission for vaccination recommendations in Iraq. In the present study, we presented the cases of infection by influenza A or B viruses. To test influenza virus types A (H1N1 and H3N2) and B, 1,359 throat and nasal swabs were collected from patients with ILI or SARI. Ribonucleic acid was extracted and amplified using a set of primers and probes. The frequency rates of infection were obtained at 1,616 (45%) and 1974 (55%) in females and males, respectively. The mean age of the participants was estimated at 31.71±22.68 with a minimum and maximum ages of 1 month and 96 years, respectively. It was revealed that influenza virus type A was the most predominant with an incidence of 16.2%, followed by type B with 0.33% incidence. It was also found that December was the most prevalent month of being infected by influenza viruses types A and B (30.02% and 0.48%, respectively). Vaccination in September would likely protect the highest number of patients. It was clear that the influenza A virus was predominant over type B. In Iraq, influenza A and B viruses were found in a large percentage of ILI and SARI cases. Additionally, males were reported to be more likely to become infected than females.

    Keywords: Real-time RT-PCR, Influenza type A, Influenza type B
  • E. N .Morozova *, V. N .Morozov, A. V. Tverskoi, S. N .Perepelkina, V. P .Konshina Pages 879-886

    Diffuse nodular lymphoid hyperplasia is a rare gastrointestinal disease that can be diagnosed by multiple nodules in the small intestine, large intestine, or both. Immunodeficiency and infections are the common situations that lead to the diffusion of nodular lymphoid hyperplasia. For instance, Giardia lamblia and Helicobacter pylori are the major pathogens leading to this disorder. Diffuse nodular lymphoid hyperplasia leads to allergic reactions, immunodeficiency, and autoimmune diseases. Imunofan-RDKVYR Peptide-is a potential agent in regenerative medicine. The present study aimed to investigate morphological features of the aggregated lymphoid nodules of the small intestine after the Imunofan (IM) administration following Cyclophosphamide-induced immunosuppression. In total, 72 Wistar male rats were randomly divided into two groups (n=36). Group I was considered the control group, and group II was subjected to intramuscular injections (needle 21 G) of0.2 ml of normal saline following the Cyclophosphamide-induced immunosuppression on the2nd, 4th, 6th, 8th, and 10th days of the experiment. The animals in group II were injected with Cyclophosphamide at a dose of 200 mg/kg bodyweight to induce immunosuppression. The animals in the experimental group (n=36) were subjected to intramuscular injections (needle 21 G) of the 0.2 ml IM at a dose of 0.7μg/kg body weight on the 2nd, 4th, 6th, 8th, 10th days of the experiment. The results of the study indicated that on the 7th day in group II, the length and width of the aggregated lymphoid nodules increased, as well as the height and width of the lymphoid nodules and internodular zones as structural components of the lymphoid formations in the small intestine. In group I, by the 30th day of the experiment, the linear dimensions of the aggregated lymphoid nodules exceeded, but to a lesser extent than on the 7th day of the experiment which explains the ability of IM to neutralize the effects of Cyclophosphamide.  It should also be noted that the IM was performed to regenerate damaged cells which helped maintain the population of lymphocytes in the limb and led to an increase in linear dimensions (length and width) not only between the joint but also in the lymph nodes.

    Keywords: Aggregated lymphoid nodules, cyclophosphamide, Imunofan, rats, Small intestine
  • F. A .Alwaeely *, K. N. Madlum, M. A. Alsaadi Pages 887-894

    Immune balance during infection is critical for both supporting the defense of the immune system of the body and preventing an overly aggressive immune response. Foxp3, a transcription factor of regulatory T cells, plays a critical role in balancing the immune system of the body. Propolis has been shown to affect Foxp3 expression. This study aimed to verify the effect of propolis extracts on in vitro Foxp3 gene expression in peripheral blood mononuclear cells (PBMCs) stimulated with Pseudomonas aeruginosa Ag. In this study, a total of 20 apparently healthy volunteers were included, with 10 males and 10 females within the age range of 20-40 years old. Five ml of blood were drawn from each participant to assess Foxp3 gene expression in PBMCs using density gradient lymphoprep and stimulated with P.aeruginosa lipopolysaccharide (LPS) in vitro. The samples were divided into four distinct groups as follows: LPS stimulated PBMCs, ethanol-extracted propolis (EEP) + LPS stimulated PBMCs, and water-extracted propolis (WEP) + LPS stimulated PBMCs and PBMCs as the control group. The Foxp3 gene expression level was estimated in all four groups following a period of 48 h of cultivation by real-time polymerase chain reaction technique using SYBR green dye. Results of the study indicated that propolis had a great effect on the mRNA Foxp3 expression. Both EEP and WEP had immunomodulatory effects through the Foxp3 mRNA expression, both the EEP and WEP could significantly inhibit Foxp3 mRNA gene expression by human PBMCs after stimulation with pseudomonas Ag in vitro. Propolis exhibited an immunoregulatory effect which was the same with ethanol and water extracts on Foxp3 mRNA gene expression.

    Keywords: Foxp3, Gene expression, Immunomodulatory, Propolis
  • A. S .Mohammed *, A. A .Al Janabi Pages 895-901

    Interleukin-17A (IL-17A) is a member of the Interleukin-17 family, which belongs to the pro-inflammatory cystine-knot cytokines. Recent studies on the etiology of breast cancer have focused on the role of immunity and inflammation. The pro-inflammatory cytokines IL-17A can medicate cancer-related inflammation. The present study aimed to analyze the mutation in physicochemical properties and structure of the Interleukin-17Agenein developing breast cancer using bioinformatics methods. A total of 60 blood samples were obtained from Iraqi women aged 25 to 75 with breast cancer. Twenty blood samples were obtained from healthy women in the same age range as a control group. Deletion and missense mutations detected by BLAST in samples with breast cancer. The present study determined the physicochemical properties of IL-17A such as hydrophilic nature, alpha-helical and 3D structure. The results of this study indicated that IL-17Ais considered a marker for a patient with breast cancer. Also, mutations in the IL-17Agene affect the structure and physicochemical properties of the IL-17A protein complex.

    Keywords: Bioinformatics, Breast cancer, IL-17A
  • A. Hajizadeh, S. M. Abtahi Froushani *, A. A.Tehrani, S.Azizi, S. R. Bani Hashemi Pages 903-912

    Naringenin is one of the most important and abundant known flavonoids found in grapefruit and other citrus fruits. This experimental study aimed to assess the clinical effects and immune responses of naringenin in the animal model of rheumatoid arthritis (RA) according to various reports on its anti-inflammatory effects and modulation of the immune system. To this end, 40 Wistar rats in the weight range of 160-180g were randomly assigned to four groups (n=10) including healthy, control, naringenin, and methotrexate orally treated groups. To induce RA disease, a compound of 200 μl of Freund's adjuvant and collagen type II was injected subcutaneously into the rear footpads of rats. The severity of RA clinical signs was assessed based on a standard scoring method. The treatment lasted for three weeks (days7-28 after induction). The obtained data pointed out that the levels of C-reactive protein (CRP), myeloperoxidase, nitric oxide, IL-17, and IFN-γ cytokines significantly increased in the RA rats, while the level of their serum antioxidants significantly reduced, compared to the healthy rats. The inflammation of the paws and the level of CRP decreased similarly in both methotrexate and naringenin-treated groups. In the naringenin-treated group, a further decrease was detected in serum myeloperoxidase, nitric oxide, and the total antioxidant capacity occurred, as compared to the methotrexate-treated rats. Nonetheless, IL-17 and IFN-γ cytokines levels were further decreased in the methotrexate-treated group. Accordingly, it can be concluded that naringenin can be effectively used for the reduction of inflammatory effects and control of RA disease.

    Keywords: Rheumatoid arthritis, Naringenin, Methotrexate, Wistar rats
  • M. Faeqali Jan *, H .Muneer Al Khafaji, B. Hasan Al Saadi, M. K. Aneed Al-Saedi Pages 913-923

    It has been approved that neutrophils are responsible for many inflammatory lung diseases, such as acute respiratory distress syndrome, chronic obstructive pulmonary disease, and asthma. It is well documented that the CXC chemokine interleukin-8 (IL-8) plays a key role as a potent neutrophil recruiting and activating factor. Asthma is one of the most common major non-contagious diseases and has a substantial impact on the patient’s quality of life. The current evidence suggests that asthma is a complex multifactorial disorder, and its etiology is increasingly attributed to interactions between genetic susceptibility, host factors, and environmental exposures. IL-8 plays an important role in respiratory diseases and is a known regulator of pulmonary inflammation and immunity, induced phagocytosis, and promoted angiogenesis. This study aimed to investigate the IL-8 gene expression in blood samples of bronchial asthma patients. Therefore, the blood samples were taken from two groups of participants, including the group of patients with asthma (n=100) in the age range of20-61years and the group of healthy individuals (n=50).The obtained results indicated that the expression of IL-8 mRNA in the group of asthma patients was three times higher than that in the group of healthy individuals. Therefore, it is suggested that the antagonism of IL-8 could be a potent therapeutic strategy in the treatment of asthma.

    Keywords: Bronchial asthma, Gene expression, IL-8, qRT-PCR, RT-PCR
  • Y .Sabah Abdulameer *, H .Hamzah Ajeel, Z .Bakir Al-Hilli Pages 925-934

    Brassica juncea (B. juncea) is an erect, and often an unbranched plant that belongs to the family Brassicaceae. The plant’s seeds have been used in many countries as a folk remedy to treat considerable common and chronic diseases. The current study aimed to investigate the possible effects of B. juncea seed extract supplementation in the drinking water as an alternative antibiotic growth promoter on poultry production. In a completely randomized design, 308 unsexed Ross broilers were allocated into 4 treatments with4 replicates, and each replicate was run on10 birds. Aqueous B. juncea seeds extract (MSE) was administered to drinking water at levels of 0, 3, 5, and 7 ml/liter to T1, T2, T3, and T4, respectively, from day 1to day 35. No significant effects were reported regarding jejunum villi height and villi thickness (P≥0.05). However, the ratio of villus height to crypt depth was increased (P<0.05), and the crypt depth was reduced (P<0.05) in birds that had been fed B. juncea seeds extract, compared to control treatment (T1) at 35 day. The B. juncea seeds extract (MSE) at the level of 7 ml (T4) yielded the highest serum total protein, phosphorus, and calcium. The T2, T3, and T4 had the lowest values of cholesterol (160, 180mg /L) and the highest value (P<0.05) of alkaline phosphatase. On day 35, the birds receiving different levels of B. juncea seed extract had lower total aerobic bacteria counts in the ileum, compared to birds fed with control treatment. The administration of B. juncea seeds extract at 3, 5, and 7 ml levels can be added to drinking water to improve gut morphology, blood biochemical traits, and intestinal bacterial load.

    Keywords: Brassica juncea Seed, broilers, Intestinal health
  • R .Farrokhi, V.Babapour *, M. Zendehdel, A. Asghari, H. Gilanpour Pages 935-948

    The present study aimed to identify the role of dopaminergic and cannabinoidergic systems in the ghrelin-induced hypophagia among meat-type chickens. In the first experiment, intracerebroventricular (ICV) injection was applied to birds with control solution, D1 receptor antagonist (5 nmol), ghrelin (6 nmol), and D1 receptor antagonist plus ghrelin. The second to sixth experiments were similar to the first one, with the difference that D2 receptor antagonist (5 nmol), D3 receptor antagonist (6.4 nmol), D4 receptor antagonist (6 nmol), the precursor of dopamine (125 nmol), and 6-hydroxy dopamine (150 nmol) instead of D1 antagonist were injected into the broiler chickens. In experiment 7, control solution and different levels of ghrelin antagonists (5, 10, and 20 nmol) were injected. In experiment 8, the chickens were ICV injected with control solution, ghrelin antagonist (10 nmol), dopamine (40 nmol), and ghrelin antagonist plus dopamine. In experiments 9 and 10, CB1 and CB2 receptors antagonist (6.25µg and 5µg) were co-injected with ghrelin (6 nmol), respectively, measuring the food intake for 120 min after the injection. It was observed that ghrelin ICV injection considerably reduced food intake, whereas ghrelin antagonist increased food intake, depending on the dose (P<0.05). In addition, ghrelin-induced hypophagia was significantly attenuated by D1 receptor antagonist and 6-hydroxy dopamine (P<0.05), while the dopamine precursor considerably elevated the ghrelin-induced food intake (P<0.05). The dopamine-induced feeding behavior was diminished by the co-administration of [D-Lys-3]-GHRP-6 (10 nmol)+dopamine (40 nmol) (P<0.05). In addition, CB1 receptor antagonists enhanced the ghrelin influence on food intake (P<0.05). The results implied that the hypophagic impact of ghrelin was probably mediated by D1 and CB1 receptors within neonatal broilers.

    Keywords: Ghrelin, Cannabinoid, Dopamine, Broiler chicken, Food intake
  • A .Abdulaali Azeez, E. Mohammed Mustafa *, O. Mahrouf Ali Shoshin Pages 949-955

    Green tea (GT) is believed to have antioxidant properties and beneficial effects on the treatment of some diseases. However, few findings were found concerning the impact of GT on oxidative stress. In the present study, the protective influence of GT against the oxidative stress caused by hydrogen peroxide (H2O2) in rats was evaluated. The research groups included a control (Con) group and five groups supplemented with 10g GT(G1), 20g GT(G2), 1% H2O2(P), 1% H2O2and10g GT (GP1), as well as 1% H2O2 and 20g GT(GP2). The effects of GT and H2O2 administration on serum biochemical parameters, such as lipid profile, malondialdehyde (MDA), and oxidized low-density lipoprotein (Ox-LDL) were assessed. The findings of this research revealed that the usage of GT lowered the level of cholesterol, triglyceride, LDL, MDA, Ox-LDL and coronary risk index. Moreover, an increase in high-density lipoprotein and very-low-density lipoprotein (VLDL) was observed in subjects who received GT, compared to the rats of the P group. The baseline lipid profile and GT consumption with or without H2O2 were the same between the Con and GT-treated groups. Therefore, GT usage was found to be advantageous in reducing Ox-LDL and lipid peroxidation in rats. These results confirm the traditionally claimed benefits of GT for protection against lipid peroxidation and atherosclerosis.

    Keywords: Camellia sinensis, green tea, Lipid peroxidation, OX-LDL, rat
  • K .Rasheed *, I. Thamer, F. A. Hussine, A. Ibraheem Pages 957-964

    The pancreas is a pear-shaped flat organ resembling the letter L, and yellowish to pink in color. This organ is of medical significance since it is associated with two life-threatening diseases including diabetes mellitus and pancreatic cancer. This study was conducted on male rabbits which were assigned into 3 age groups (6-month-old, 1-year-old, and 3-year-old rabbits). Physiological and histological changes of the pancreas were studied in the adopted age groups. The physiological aspect and the histological structure of the pancreas were also studied by the analysis of the level of pancreatic gland hormones and hormonal changes. Based on the results, there were significant differences in the concentration of pancreatic gland hormones. Insulin level in the second study group was more than that in the first and third groups, while the highest concentration of blood sugar (glucose) was observed in the third group, compared to the first and second. Although the basic structure of the pancreas was similar in all samples, changes were observed in the tissue structure of the pancreas throughout the process of aging. By the increase of age (from 1 to 3 years old), Langerhans islets increased in size, contained alpha and beta cells that were surrounded by a loose connective tissue in the third stage. Moreover, no significant difference was observed in the diameters of cells that produced enzymes at all stages of life. Physiological and histological changes indicated that age plays a role in the function and structure of the pancreas gland during different stages of life. In addition, this study indicated that the hormonal variability of the pancreas is closely related to the histological composition of gland components. Therefore, further studies on the role of factors, such as gender, different breeds, or environmental conditions seem to be necessary and may provide more information on factors that may affect the effectiveness and activity of the pancreas gland.

    Keywords: Age Progress, Langerhans Islets Diameters, PANCREAS, Rabbit’s Pancreatic Gland
  • S. G .Gorelik *, S. A. Kolesnikov, A. A. Zheludev, Y. A .Parkhisenko, V. V. Bulynin Pages 965-974

    Gastroduodenal bleeding is one of the most challenging issues in surgery nowadays. The crude mortality rate due to severe blood loss in gastrointestinal bleeding is very high. Therefore, the use of medications to prevent severe blood loss and protect cells from the harmful effects of hypoxia should be the focus of attention in these conditions. This experimental study was carried out to establish changes in blood parameters and humoral immunity in rats after intestinal anastomoses combined with acute blood loss after the application of catholyte and anolyte. The study included 45 male Wistar rats weighing 290-320 g that were divided equally (15 animals per group) into three groups of intact animals (group 1), animals exposed to the small bowel resection (1.5 cm) with the end-to-end anastomosis and simulated acute blood loss (group 2), and animals exposed to the small bowel resection with end-to-end anastomosis and simulated acute blood loss that were daily given catholyte in the postoperative period group (group 3). After the surgery, the rats were given a catholyte solution per os, and the operative wound was treated with an anolyte. The blood samples and the wall of the small intestine in the anastomotic zone were used as a biological substrate to study the effect of catholyte on changes in the body during the healing process. The experiment was conducted for 15 days, and the data were recorded in two intervals on the 5th and 15th days after starting the experiment. The analyzed materials evidenced that the use of catholyte and liquid with negative oxidoreduction potential (ORP) (minus 500-520 mV) resulted in positive changes in the blood cell count, humoral immunity, and phagocytic activity impaired after the small bowel resection and blood loss. The use of an anolyte disinfectant (liquid with positive ORP+710-770 mV) prevented bacterial contamination of the surgical wound. The obtained findings proved that the catholyte had a positive effect on humoral immunity and healing processes in the anastomotic zone. Furthermore, the anolyte prevented the development of purulent complications and inflammation in the area of the surgical wound, and therefore, promoted the healing processes.

    Keywords: Anolyte, Catholyte, Immunity, Intestinal anastomosis, Oxidoreduction potential (ORP)
  • S. M. J.AL Hadrawy *, Z. S .Mahdi AL Turfi Pages 975-983

    Proton pump inhibitors (PPIs) are a group of medications effectively used to inhibit gastric acid secretion and to treat many acid-related disorders, including gastroesophageal reflux disease and other gastric disorders. Recent studies recommended that they may be associated with the risk of chronic kidney disease and liver disease. Therefore, the current study aimed to investigate the effect of long-term treatment with PPIs on kidney and liver function in laboratory rats. Fifteen female albino white rats (Rattusnorvigicus) were randomly assigned to three groups of five animals. The control group was fed regular pellet, group PPI-2 received standard pellet diet and was given esomeprazole (10 mg/kg b.w.) via daily oral gavage in mornings for two weeks, and group PPI-3 was fed standard pellet diet and was given esomeprazole (10 mg/kg b.w.) via daily oral gavage in mornings for three months. Blood samples were taken after 2 weeks and 3 months by cardiac puncture for measuring serum creatinine, urea, total bilirubin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP).  In addition, kidney and liver tissues were histopathologically evaluated.  Serum creatinine, urea, ALT, total bilirubin, and ALP significantly increased in group PPI-3, compared to other groups. Histopathological study of the kidneys and liver revealed normal histology structure in the control group and the rats of the PPI-2 group, while some histological changes were observed in the liver and kidney of the animals in the PPI-3 group. The histological changes included the widening of Bowman's space and shrunken glomeruli, whereas the renal tubules had congested tubular cells. Furthermore, congestion in the blood vessels and hepatic cells degradation were observed in the liver. These data indicate that the long-term administration of PPIs has adverse effects on the structure and function of the kidney and liver.

    Keywords: Proton Pump Inhibitor, Liver Function, Kidney Function, Long-term Treatment, Esomeprazole
  • N. Nesterova *, E. Patrakhanov, V. Pokrovskii, I. Pirozhkov, T. Pokrovskaya, N. Levit, M. Ivanova, S. Kizi, A. Nesterov, V. Shutov, Y. Hoshchenko Pages 985-994
    Stroke or ischemia is caused by a blockage in a specific blood vessel that partially or completely reduces the blood flow to the brain. Nutritional factors such as antioxidants and healthy eating patterns are important variables in preventing stroke. Molecular composition properties such as molecular binding and screening can be used to evaluate the specific activity and morphological changes. The present study aimed to evaluate the effectiveness of pharmacological correction of the consequences of a hemorrhagic stroke in rats with a new derivative of taurine magnesium-bis-(2-aminoethanesulfonic)-butadioate. The animals (n=170) were divided into four groups as follows: 1) control group (n=20), 2) group 2 suffered a hemorrhagic stroke without pharmacological correction (n=50), 3) group 3 (n=50) underwent simulation of hemorrhagic stroke received Taurine at the dose of 50 mg/kg, 4) Group 4 underwent simulation of hemorrhagic stroke with correction of hemorrhagic stroke with magnesium-bis-(2-aminoethanesulfonic)-butadioate at the dose of 150 mg/kg (LKHT 3-17) (n=50). Hemorrhagic stroke was induced by transfusing autologous blood into the parietal lobe of the right hemisphere of the brain. Lethality, neurological status, locomotor, and exploratory behavior, as well as the morphological pattern of the brain damage, were assessed on the 1st, 3rd, and 7th days after the pathology simulation. Neurological deficit was determined in animals by the McGrow stroke index scale. The locomotor and exploratory behavior was evaluated using the Acti-track software and hardware complex. Two criteria were considered when assessing morphological changes in the brain: the average thickness of the cerebral cortex (in micrometers) and the number of neurons without degenerative changes. LKHT 3-17 (150 mg/kg) and taurine (50 mg/kg) reduced lethality by 1.7 and 1.36 times, respectively, on the 3rd day after stroke compared to that of the control (p<0.05). In parallel, a neurological deficit was effectively corrected LKHT 3-17 and taurine to 5.3±0.8 and 6.5±0.9, respectively, on the 1st day in contrast to the control of 8.1±0.7 points. The locomotor and exploratory behavior was significantly different on the 7th day and was accompanied by a significant increase in total activity under the influence of LKHT 3-17 to 491 conventional units (CU) compared to the control of 110 conventional units. On the 1st day, the thickness of the cortex was 1943.7±44.08 µm, and 1491.0±38.61 µm in the control and LKHT 3-17 groups, respectively. The number of neurons without neurodegenerative changes prevailed in LKHT 3-17 group (18.7±4.32), and the lowest number was observed in the group without pharmacological correction of the pathology (14.3±3.78). The taurine derivative magnesium-bis-(2-aminoethanesulfonic)-butadioate, which is a combination of the amino acid,  magnesium ion, and succinic acid, decreases the neurological deficits, lethality, and enhances the locomotor and exploratory behavior in experimental hemorrhagic stroke in rats. The effect of the studied medication on the dynamics of molecular pathophysiological mechanisms occurring in the cell requires additional research.
    Keywords: Hemorrhagic stroke, Neuroprotection, Taurine
  • A. S Netrebenko *, V. V Gureev, M. V Pokrovskii, A. V Gureeva, Y. M Tsuverkalova, I. S Rozhkov Pages 995-1004
    Chronic kidney disease (CKD) or acute kidney injury (AKI) causes impaired kidney function, leading to cognitive impairment, neuropathy, and cerebrovascular disease. Due to kidney damage, toxins stay in the blood rather than leaving the body through the urine, and brain function is affected by kidney-brain interaction. The present study aimed to investigate the protective effects of erythropoietin mimetic peptide (pHBSP) and infliximab on ischemic renal reperfusion injury. The experiment was performed on 70 white male Wistar laboratory rats which received recombinant erythropoietin, pHBSP, and infliximab. Under anesthesia, traumatic vascular clamps were applied to the left renal pedicle for 40 min, and nephrectomy was performed on the right. Functional tests and laboratory tests were performed 5 min and 24 h after the reperfusion.  Thereafter, 24 h after the surgery, the plasma creatinine and urea levels in the sham-operated animals were obtained at 45.9±0.8 mmol/L and 6.7±0.2 mmol/L, respectively. Plasma creatinine and urea levels in the control group animals were 102.63±3.6 mmol/L and 21.80±1.29 mmol/L, respectively. The administration of pHBSP and infliximab to the animals with ischemia-reperfusion kidney injury has a pronounced nephroprotective effect, as compared to erythropoietin. There was a significant decrease in blood levels of creatinine and urea, improvement of microcirculation in the kidney, normalization of glomerular filtration rate, and fractional sodium excretion. The results of the study demonstrated pointed to the prospects of pHBSP and infliximab administration in ischemia-reperfusion kidney injury and justified the feasibility of further research in this field.
    Keywords: Erythropoietin mimetic peptide (pHBSP), Infliximab, Ischemia-reperfusion kidney injury, rats, Microcirculation
  • A. Agarkova, A. Tverskoi *, V. Morozov, A. Tverskaya, E. Morozova, T. Mukhina Pages 1005-1012

    The soft and delicate tissue of the brain, which is the center of our coordination, is protected by its surrounding layers. The disruption of these layers results in complicated situations and serious health problems. The brain has three protective layers of bone or skull tissue, the blood tissue layer, and finally the meningeal layer. The layer of blood tissue contains the blood vessels that are located between the skull and the meningeal membranes. If germs or foreign matter enter the fluid through the blood vessels under any circumstances and cause infection, the bones that protect the meninges will break and cause tissue damage. The present study aimed to assess the histological and immunohistochemical characteristics of the brain of rats that underwent induced acute purulent pneumococcal meningitis after antibiotic therapy with Ceftriaxone. A number of 20 white adult male Wistar rats were assigned to three groups. The first group (n=5) regarded as the control were injected with a saline solution into the subarachnoid space in an equivalent amount. The second and third groups of rats (n=5 and 10, respectively) were infected with acute purulent meningitis by the injection of 10 μl of Streptococcus pneumoniae (S. pneumonia) suspension into the subarachnoid space of the brain using a 23-G needle. The various areas of the brains of rats after meningitis induced by S. pneumoniae were examined after the treatment with Ceftriaxone. The S. pneumoniae culture was injected into the subarachnoid space in the area of the rhomboid fossa. Treatment started 18 h after the injection. On day 10, a repeated puncture was performed with the analysis of cerebrospinal fluid in order to confirm the absence of meningitis; thereafter, the animals were taken out of the experiment. No signs of meningitis were found on histological examination. Mild perivascular and pericellular focal edema were revealed with signs of overload of the lymphatic system in the brain and focal ischemic changes in neurons. The investigation of expression with caspase-3 revealed a positive reaction of individual neurons. A positive reaction with antibodies to NeuN and Doublecortin was detected in most neurons; moreover, Glial fibrillary acidic protein (GFAP)-positive astrocytes and their processes were visualized in all layers of the brain substance. The reaction with neuron-specific enolase (NSE), microtubule-associated protein 2 (MAP-2), CD 31, and CD 34 was negative. Typical structure and pictures pointed to an intact brain and purulent meningitis in the first and second groups. The microscopic image and the changes revealed during immunohistochemistry by dual corticosteroid antibodies and neuronal nuclear protein were characterized by predominantly cytoplasmic and perinuclear reactions, respectively. Some neurons are positive for caspase-3 and are related to changes in the characteristic of premature aging.

    Keywords: Acute meningitis, Ceftriaxone, Immunohistochemical markers, Brain of rats
  • M .Aal Aaboda *, A. R.Abu Raghif, N. R .Hadi Pages 1013-1024

    Ischemia/reperfusion injury (IRI) is caused by a sudden temporary impairment of the blood flow to the particular organ. The IRI of the kidneys is one of the main causes of acute kidney injury. A vigorous inflammatory and oxidative stress response to hypoxia and reperfusion usually happens as IRI consequences that disturb the organ function. The current study aimed to investigate the effect of antagonizing toll-like receptors (TLRs) effects by lipopolysaccharide obtained from Rhodobacter sphaeroides (LPS-RS) on this critical condition. In total, 28 adult male Wistar rats were divided into four groups (n=7) as follows: the sham group which underwent only laparotomy; control group that underwent laparotomy and IRI induction; vehicle group which was similar to the control group plus vehicle treatment, LPS-RS group that was similar to the control group but was pretreated with 0.5 mg/kg of LPS-RS. The results of the current research showed that LPS-RS reduced interleukin-1β, interleukin-6, tumor necrosis factor α, and 8-isoprostane levels, compared to the control IRI group. However, LPS-RS did not ameliorate the kidney injury as manifested by the elevated levels of urea, creatinine, and neutrophil gelatinase-associated lipocalin. Taken together, the present study demonstrated that LPS-RS at the tested dose failed to offer a renoprotective effect against the IRI in rats.

    Keywords: F2-Isoprostane, Il6, Il1β, Lps-Rs, Neutrophil Gelatinase Associated Lipocalin, Ngal, Renal Ischemic Reperfusion Injury, Tlr2, Tlr4, Tnfα
  • N. I Nesterova *, V. V Pokrovskii, E. A Patrakhanov, I. V Pirozhkov, T. G Pokrovskaya, N. B Levit, I M.I, S. N. M. Kizi, A. V Nesterov, V. I Shutov, Y Yuri A. Hoshchenko Pages 1025-1034
    Sudden loss of blood flow to an area of the brain causes ischemic stroke, which leads to the loss of nerve function in the brain. The brain tissue leads to the death of brain cells in less than a few minutes due to the lack of oxygen and nutrients. This study aimed to evaluate the effectiveness of pharmacological correction of the consequences of ischemic stroke with a new derivative of taurine magnesium-bis-(2-aminoethanesulfonic)-butanedioate under laboratory code LKHT 3-17 in rats. The ischemic stroke was simulated by electrocoagulation of the right middle cerebral artery. The assessment of lethality, neurological status, locomotor, exploratory behavior, and morphological pattern of the brain damage was carried out on the 1st, 3rd, and 7th day after the pathology simulation. Neurological deficit was determined by the McGrow stroke index scale. The locomotor and exploratory behavior was evaluated using the Acti-track software and hardware complex. When assessing the morphological changes in the brain, attention was paid to two criteria, including the average thickness of the brain cortex and the number of neurons without degenerative changes. The substances were administered 60 minutes before the start of surgery. The animals were divided into an intact group (n=20); ischemic stroke simulation group without pharmacological correction (n=50); a group with correction of the ischemic stroke with taurine at the dose of 50 mg/kg (n=50); and a group with correction of ischemic stroke with magnesium-bis-(2-aminoethanesulfonic)-butadioate (LKHT 3-17) at the dose of 150 mg/kg (n=50).LHT 3-17 (150 mg/kg) and taurine (50 mg/kg) reduced lethality by 1.55 and 1.47 times, respectively, on the 7th day after stroke, compared to the control group (P<0.05). In parallel, an effective correction of neurological deficit was found for LKHT 3-17 and taurine to 4.0±0.8 and 7.6±0.9, respectively, on the 3rd day in contrast to the control of 8.1±0.8 points. The locomotor and exploratory behavior was most significantly different on the 1st and 7th days and was accompanied by a significant increase in the speed of movement under the influence of LKHT3-17 to 20 and 20 conventional units, compared to the control of 7 and 5 cu. On the 1st day, the thickness of the cortex was 1877.3±43.3 µm in the control group, and 1531.8±39.1 µm in the LKHT 3-17 group. The number of neurons without neurodegenerative changes prevailed in the group administered with LHT 3-17 (19.3±4.3), and the lowest number was observed in the group without pharmacological correction of the pathology (14.3±3.7).LKHT 3-17 at a dose of 150 mg/kg is more effective than taurine 50 mg/kg in protecting nerve activity in experimental ischemic stroke and reducing lethality, minimizing nerve defects, reducing volume, accelerating the process of tissue repair, helping stroke, and activating the regenerative processes.
    Keywords: Ischemic Stroke, Neuroprotection, Taurine
  • S. Rasheed, L. Younis *, Q. Aboud Pages 1035-1045

    The present study aimed to assess the relationship of Growth Differentiation Factor 9 (GDF9) genotypes with calving rate, Follicle-stimulating hormone (FSH), and Estradiol (E2) in the Iraqi Holstein-Friesian breed. A number of 15 blood samples were collected from a mother of dizygotic twin birth (DZTB) (with high calving rate records), and another blood sample was collected from 15 single birth (SB) cows. The DNA was extracted and six primers were designed for PCR and sequencing analysis. The FSH and E2 levels were tested through the estrus phase for the two groups (n=10 in each group). The sequence evaluation revealed the presence of two single nucleotide polymorphisms (SNPs) in exon II: A (1109) T and G (1133) A. The genotypic frequency for mutant genotypes was higher significantly (P<0.01) in DZTB cows (with calving rate), as compared to wild genotypes at the same loci. On the other hand, the wild genotypes recorded a significant increment (P<0.01) for SB cows, when compared to mutant genotypes in the same loci. Moreover, a significant rise (P<0.05) was reported in E2 and FSH levels for DZTB cows and mutant genotypes (P<0.01) against SB cows and wild genotypes in 0 and 24 h of estrus phase, respectively. Furthermore, non-significant differences were recorded in E2 concentration among the same genotypes at the same period. In conclusion, the GDF9 exon II SNPs increased the calving rate in Holstein-Friesian cows. The blood FSH and E2 concentrations were higher in the DZTB cows and control the superovulation. Finally, these SNPs can be regarded as markers to accelerate the breeding programs and used in embryo transfer and in vitro embryo production for Iraqi Holstein-Friesian cow breed.

    Keywords: Calving rate, Dizygotic twinning, GDF9 polymorphism, FSH, Estradiol
  • A .Nouri *, M. Bashashati, S. Gh. Mirzaie, A .Shoshtari, M. Banani Pages 1047-1053

    Avibacterium (Haemophilus) paragallinarum (Av. Paragallinarum) is the causative agent of Infectious Coryza (IC) in chickens. Despite the worldwide distribution of IC, no systematic study, to the best of our knowledge, was conducted on isolation and characterization of Av. Paragallinarum in Iran. The present study aimed to isolate and perform antibacterial susceptibility testing (AST) of IC agents from suspected backyard chickens with typical symptoms of IC in avian markets. From 18 collected choanal swab samples, four (22%) isolates of Av. Paragallinarum were detected by culture methods based on satellite growth on blood agar, which was confirmed by the biochemical reaction of Catalase and Oxidase tests and species-specific PCR (HPG-2). The hypervariable region of the hemagglutinin genes of 4 isolates was amplified and obtained sequences were deposited at a gene bank for more characterization. Meanwhile, 12 (66%) positive reactions were detected by observing expected 500 bpb and using PCR (HPG-2) on swab samples. Antibiotic susceptibility testing (AST) of obtained isolates were analyzed using the Kirby-Bauer disk diffusion method on Columbia agar with horse blood. Isolates were found to be resistant to amoxicillin, oxytetracycline, streptomycin, trimethoprim/sulfamethoxazole (up to 75%) and sensitive to cefalexin, ceftriaxone, enrofloxacin, florfenicol, gentamycin, linco-spectin, neomycin, doxycycline (50%), danofloxacin (75%), flumequine (50%), ofloxacin (75%). An intermediate growth inhibitionzone has been observed around antibiotic discs for ampicillin, colistin, erythromycin, penicillin, tiamulin (75%), tylosin (75%). In summary, to the best of our knowledge, this study is the first report of isolation and identification of Avibacterium paragallinarum from backyard chickens which may be a source of IC for commercial chicken flocks. Moreover, the prevalence of resistance to some antibacterial drugs of IC agents may impose an additional threat to the poultry industry. A more in-depth study is recommended to develop a low-cost autogenous IC vaccine for small-scale flocks of poultry to prevent and manage the disease and establish antimicrobial resistance.

    Keywords: Infectious Coryza, Avibacterium paragallinarum, PCR, Isolation, Antibiotic susecptibility test
  • R. Naji Hasan *, S .Abdal Kareem Jasim Pages 1054-1059

    A gram-positive bacterium, Staphylococcus aureus, which is widely distributed is considered as a bacterial infection that commonly infects the skin and mucous membranes. Such infections can be the cause of death and illness. In the present study  by using reverse transcription-polymerase chain reaction (rt-PCR) the Panton-Valentine leukocidin (PVL) and mecA genes of S. aureus which were isolated from skin and soft tissue infections (SSTIs) in Baghdad, Iraq were investigated. This study included 96 S. aureus isolated from SSTIs and identified by Vitek. The results showed that 61 (63.5%) and 48 (50%) of the isolates were positive for PVL and mecA genes, respectively. This work presented an effective real-time PCR technique for detecting PVL genes alone or in conjunction with mecA. The rt-PCR allows for easier reaction monitoring and eliminates the need for post-PCR processing, saving both resources and time. Moreover, it is ideal for diagnostic applications because of its high sensitivity, simplicity, and specificity. Besides, the rt-PCR has an option to do all the procedures in an automated mode of action.

    Keywords: MeC A gene, PVL gene, Real-time PCR, Staphylococcus aureus
  • S .Ahmed Hasan *, T .Fakhraddin Raheem, H. Mohammed Abdulla Pages 1061-1067

    Klebsiella pneumoniae is globally responsible for hospital- and community-acquired infections. This study aimed to determine the prevalence of K. pneumoniae and investigate the antibiotic resistance profile among clinical specimens at Azadi Teaching Hospital in Kirkuk, Iraq, and detect the rpoB gene for molecular identification of  K. pneumoniae in comparison with phenotypic and biochemical methods. In total, 250 clinical specimens were collected from patients in Azadi Teaching Hospital in Kirkuk, Iraq, between January 2018 and May 2018. The isolates were identified by morphologic and biochemical testing. Kirby-Bauer disk diffusion method was used in the antibiotics susceptibility test. Following that, 19 (7.6%) K. pneumoniae isolates were isolated from 250 clinical specimens (5 [5.61%] and 14 [8.69%] from males and females, respectively), and most of them (n=12; 11.76%) were isolated from the age group of 10-35 years old. The isolates were reported high resistance towards various types of antibiotics, especially penicillins and cephalosporins. In contrast, K. pneumoniae showed very low resistance to imipenem and amikacin (5.26% and 10.52%, respectively). The range of multidrug-resistant K. pneumoniae isolates in this study was estimated at 100%. In gene detection, all isolates in this study showed PCR product with 108 bp by K. pneumonia specific primer (rpoB). Developed antibiotic policies and regular surveillance of antibiotic susceptibility patterns may help to overcome the indiscriminate use of antibiotics that is a major cause of the emergence of drug resistance among pathogens.

    Keywords: K. pneumoniae, Multidrug Resistance, Rpob Gene
  • N. I. Zhernakova *, S. S .Bunova, N. M .Agarkov, D. T. Lebedev, V. V. Aksenov Pages 1069-1076

    Cardiovascular diseases are among the most common causes of disability and death in the world, and the number of patients with this category of diseases is increasing every year. This study aimed to investigate the role of vitamin D and the problems caused by its deficiency on the cardiovascular system. Level of D-(25[OH]D) in blood was studied by enzyme immunoassay in 95 elderly patients with myocardial infarction (MI) (the main group) and 92 elderly patients with no history of MI. The level of lipid metabolism as an indicator was determined using a KoneLab 300 auto-analyzer. Based on the results of this study, it was found that the elderly group without MI had the highest amount of D-(25[OH]D)(24.5±1.2), compared to the elderly group with MI (14.8±1.3). The rates of expressed deficiency, deficiency, and insufficiency in the group of elderly with MI were 53.6±5.1, 23.2±4.4, and 12.6±3.4%, respectively. This experiment has shown that D-(25[OH]D) is involved in lipid metabolism and reduces the accumulation of cholesterol by macrophages. The content of vitamin D in blood plasma was a prognostic predictor of MI, which improved MI in the elderly. Regardless of pathological changes, the deficit level of D-(25[OH]D) should be considered a laboratory predictor of MI in the elderly.

    Keywords: lipid metabolism, Myocardial infarction, Old age Prognostic value, Vitamin D
  • N .Sabah Younus, Z. Abdul Munim Sharba *, M .Fakhry Altaee Pages 1077-1085

    miRNAs regulate protein abundance and control diverse aspects of cellular processes and biological functions in metabolic diseases, such as obesity and diabetes. Lethal-7(Let-7) miRNAs specifically target genes associated with diabetes and have a role in the regulation of peripheral glucose metabolism. The present study aimed to describe the gene expressions of the let-7a gene with the development of diabetes in Iraq and the difference in the expression of this gene in patients with diabetes and healthy individuals. The association between age and gender with the development of diabetes was studied in this study and the results were compared with those of healthy individuals in the group of control. Based on the obtained results, there was a lack in the mean of gene expression level (ΔCt) in patients, compared to controls. Moreover, the gene expression folding (2-∆∆Ct) of the let-7a reflects significant differences in terms of gene expression between groups of patients and controls, and the level of let-7a expression was reported to be 12.97 in patients with diabetes. On the other hand, significant difference was observed in terms of age and gender between diabetic patients and controls. The findings suggest that diabetes can affect individuals in all age groups and occur regardless of gender in both males and females. Based on the obtained results in this study, the gene expression level of miRNA let-7a was lower in diabetic patients compared to healthy individuals in the group of control. This also reflects differences in the gene expression fold (2-∆∆Ct) of gene let-7a between both groups of patients and controls.

    Keywords: Gene expression, diabetes, Micrornalet-7a
  • M. Y. Skorkina *, T. S. Shevchenko, A. S. Taranenko, E. S .Shentseva, L. R .Zakirova Pages 1087-1093

    Acute lymphoid leukemia (ALL) affects the lymph cells or lymphocytes that make up the lymph tissue and prevents the proper maturation of the bone marrow cells.  The processes through which cells convert mechanical stimuli into biochemical signals are called mechanical transitions and result in the sensation of specific cellular responses. In the present study, the functional properties of granulocytes of the patients with ALL were investigated using the in vitro mechanical stress model. The experimental part of the work was executed using blood from patients with ALL (n=30) being treated in the Hematological Department of Belgorod Region Hospital, Belgorod, Russia. The patients were in the age range of 18-45 years. Sample blood was obtained from all the patients who underwent a standard course of chemotherapy. Blood sampling was performed using a venepuncture and collected into the vacuum tubes Vacuette K3E. Blood samples from each experimental group were divided into two groups of control and experiment. The injection model of mechanical stress was used for the experiment group in vitro. Subsequently, the adenosine triphosphate (ATP) concentration increased by 1.8 times in this group, compared with the controls. Young’s module, which numerically characterizes the rigidity of the granulocytes’ plasmalemma, decreased by 54.4% (P<0.05) under the influence of mechanical stress. The surface potential of plasmalemma was not significantly different between samples in the group of control and experiment in patients with ALL. However, the adhesive force between erythrocyte and granulocyte increased by 30.7% (P<0.05).  The osmotic load test showed an increase in the cell’s volume during incubation. The use of membrane reserve by granulocytes increased by 47% (P<0.05) at the initial seconds of incubation. The obtained results pointed to the regulatory role of ATP molecules in intercellular signaling and add to the present literature regarding the mechanisms of intercellular interaction in the microvasculature on the development of leukemia. Moreover, the obtained results can be taken into account for the development of new pharmacological immune correctors.

    Keywords: Adhesive properties of biomembranes, Atomic Force Microscopy, Granulocytes, Potential surface, Young’s module
  • E. Riyadh Mohsen, N. H .Ali *, H. A .Aldaoseri Pages 1095-1105

    This study aimed to determine the correlation of disease activity of rheumatoid arthritis (RA) with Th17/regulatory T cell (Treg) and Forkhead box protein 3 (Foxp3) cells ratio in patients under therapy with anti-tumor necrosis factor (TNF)-α. Totally, 84 patients with RA and 13 healthy controls were included in this case-control study. The patients were divided into four groups to receive only methotrexate (MTX) (n=25), monotherapy (anti-TNF) (n=18), and combined therapy (MTX+anti-TNF) (n=26); however, one group received no medications (n=15) and was regarded as a positive control. Other 13 healthy controls that were considered negative controls were also enrolled in this study. Patients with RA were attending Basrah General Hospital, Rheumatology Unit, Biological Therapy Center for receiving anti-TNF therapy. Flow cytometry was used for measuring Treg/Foxp3 and Th17 markers, and the DAS-28 score was utilized to measure RA disease activity. Anti-TNF inhibitors (e.g., infliximab and etanercept), as well as other inflammatory and hematological parameters (e.g., erythrocyte sedimentation rate, total white blood cells, lymphocytes, monocytes, and neutrophil counts), were also measured in this study. DAS-28 as a disease activity score was significantly correlated with Th17/Treg/Foxp3 ratio and the Th17 cells count. Statistically, Th17/Treg/Foxp3 ratio was not correlated with body mass index, morning stiffness, and duration of the disease. Th17/Treg/Foxp3 ratio correlated significantly with DAS-28 as an RA disease activity. The lower Treg/Foxp3 frequency led to the higher DAS score reflecting higher disease activity. In the combined therapy group, disease activity was found lower than that in other patient groups indicating the effect of this combination on the relationship between MTX and anti-TNF. This study demonstrated that the main advantage of this combined therapy in RA patients was the reversion of Th17 cell expan sion.

    Keywords: Anti-TNF, Etanercept, immunotherapy, Infliximab, Rheumatoid arthritis, Th17, Treg, Foxp3 ratio
  • T. V. Pavlova *, N. B. Pilkevich, D. V. Bessmertnyi, I. A. Pavlov, D. V .Atiakshin, L. A .Pavlova Pages 1107-1113

    Kidney malignancies are among the most deadly genitourinary tumors. It is more common in males and is often seen in people aged 60-70 years old. The incidence rate of kidney cancer seems to be increasing. One reason for this may be the fact that imaging techniques, such as computed tomography scans are more commonly used. These tests may lead to the accidental detection of more kidney cancers. Fortunately, kidney cancer is often detected in the early stages, when the tumor is small and confined to the kidney. The objective of this study was the development of new diagnostic immunohistochemical methods. Clinical examination material of 134 people, including 94 (70%) males and 40 (30%) females, were used in this study. Immunohistochemical staining of tryptase was carried out in compliance with the requirements using Anti-Mast Cell Tryptase antibodies. Goat anti-mouse antibodies #AS-M1-HRP were used as secondary antibodies, visualized with ImmPACTTM DAB Peroxidase Substrate Kit (#SK-4105) according to the instructions of the manufacturer. The nuclei were counterstained with Mayer's hematoxylin, and the sections were embedded in a permanent mounting medium. The immunohistochemical study showed an increase in both tryptase- and chymase-positive mast cells in the renal parenchyma, compared with the control group. The number of mast cells with tryptase expression directly in the tumor was significantly less than the peritumoral localization. A similar pattern was observed for chymase-positive mast cells as the content of the tumor was more than 10times higher than the intratumoral arrangement. The histological and immunological characteristics did not differ in different age groups. The immunohistochemical method of research in the diagnosis of renal tumors plays an important diagnostic and prognostic value. It can assist pathologists in difficult and ambiguous cases to correctly diagnose renal tumors. This will make it possible to prescribe the correct treatment and predict the course of malignant tumor growth in patients.

    Keywords: Kidneys, metastases, oncourology, Trace elements
  • J .Jumintono *, S. Alkubaisy, D .Yánez Silva, K .Singh, A. Turki Jalil, S. Mutia Syarifah, Y. Fakri Mustafa, I .Mikolaychik, L. Morozova, M .Derkho Pages 1115-1123

    Physical and chemical changes caused by oxidative stress in the spermatozoa membrane can reduce spermatozoa function and even lead to death. Cystamine (NH2-CH2-CH2-SH, β-mercaptoethylamine) is a natural substance that modulates the endocrine and metabolic status of animals. This substance has antioxidant and anti-apoptotic effects by inducing intracellular cysteine accumulation. Cystamine is used to treat many diseases despite its many side effects. Sheep semen is sensitive to the stressful condition of chilling storage, which restricts semen storage for artificial insemination in commercial herds. The effect of cystamine on spermatogenesis is not yet fully understood. The present study aimed to investigate the effect of cysteamine addition to the sheep sperm extender during cooling storage on semen quality parameters. Sperm samples were collected from six Edilbayevskaya rams (2 and 3 years old, 70-85 kg). The samples were diluted by extender and supplemented with different concentrations of cysteamine (0, 1, 2, 5, and 10 mM) and cooled to 4ºC for 50 h. Motility parameters, membrane integrity, viability, lipid peroxidation, and mitochondrial activity of cooled semen were evaluated at 0, 25, and 50 h of cooling storage. Although cysteamine failed to affect semen quality at start time (0 hrs), extender supplementation with cysteamine improved sperm total motility, progressive motility, and mitochondrial membrane potential during storage periods (P≤0.01). Moreover, using 1 and 2 mM cysteamine functionally and viably improved (P≤0.01) sperm membrane compared to other treatments. Antioxidant potential (AOP), lipid peroxidation (LPO), and total glutathione (tGSH) (except AOP at 50 h) were significantly different after semen storage at 4 °C. Therefore, levels of AOP and tGSH were significantly increased by using cysteamine. Cysteamine supplementation (1 and 2 mM cysteamine) leads to lower levels of LPO (p<0.01) at 0, 25, and 50 h. Therefore, finding and using the best concentrations of cysteamine in a cooling extender could be effective in saving sheep semen against damages of the cooling storage process.

    Keywords: Cooling, Cystamin, Semen, Sheep
  • A .Musaev *, S. Sadykova, A. Anambayeva, M. Saizhanova, G. Balkanay, M. Kolbaev Pages 1125-1135

    Mare's milk is a highly valuable organic substance that has a great potential to replace cow's milk. Consumption of cow's milk causes digestive disorders in some individuals. Immunoglobulin E (IgE)-mediated cow’s milk allergy (CMA) is one of the most common food allergies among infants. Therefore, finding a protein substitute with the same nutritional value is a priority. Mare's milk can be a good substitute for cow's milk, especially for those suffering from CMA. Prerequisites for this study were the recent interest in mare's milk, as an ancient relic of the Turkic peoples which contains lots of nutrients. The present study aimed to systematize relevant information on the composition of mare's milk and its application in medicine. Google Scholar, PubMed, Cochrane, Elsevier, CyberLeninka were employed for a comprehensive literature search. The searched keywords for this study were mare's milk, saumal, composition, properties, use in medicine. A total of 77 sources were selected for reviewing the literature. Most sources were in English, except for one of the bottom 40 sources published in the last 10 years. Among the milk of many mammalian species, mare's milk is chemically similar to human milk so it can be used as a substitute. It is also used to feed people with various health conditions, especially in patients at risk, or suffering from tuberculosis, hepatitis C, psoriasis, and various types of immunodeficiency. The present study describes the rich composition, antibacterial and antiviral properties of mare's milk. A review of the literature revealed that mare's milk is an excellent thirst quencher, and has valuable nutrients necessary for the human body which is by no means inferior to human milk.

    Keywords: antibacterial property, Antiviral property, Composition, Mare’s milk
  • A. M. Baqir AlDhalimy, S. K. Alabsawy, M .Al Mousaw, D. M .Al Dhalemi * Pages 1137-1142

    Stress is defined as physical and/or psychological modifications that disrupt homeostasis in living organisms. The stimuli that confront homeostasis are determined as stressors; these external factors may be physical, chemical, psychological, and environmental. The results of some studies have shown that ascorbic acid is related to fertility and has an evolutionary significant role as an essential nutrient for humans and other animal species. Selenium is the most important mineral element in protecting health and growth and performing various biochemical and physiological functions. Therefore, the present study aimed to determine the protective effects of vitamin C and selenium against restraint stress levels that caused a decrease in sperm quality in rats. This study was conducted on40 adult male Wistar rats that were randomly divided into 4equal groups (n=10 each). The first group (vitamin C group)was exposed to restraint stress for 6 h a day and supplemented with vitamin C (50 mg/kg bw/day) orally by gavage; the second group (Se group)was exposed to restraint stress for 6 h a day and supplemented with selenium (0.02 µg /kg bw/day) orally by gavage; the third group (negative control[NC] group)was exposed to restraint stress for 6 h a day and given normal saline (2 ml)orally by gavage; the fourth group (positive control [PC] group)was not exposed to restraint stress and given normal saline (2 ml)orally by gavage. The results showed that all the sperm parameters, such as total and progressive motility, and sperm viability increased significantly (P≤0.05) in vitamin C and Se groups, compared to the NC group. The rate of acrosome defects in vitamin C, Se, and PC groups was significantly reduced (P≤0.05), compared to the NC group. Moreover, the findings showed no significant differences among all the four groups. The results of the current study confirmed the ameliorated effect of vitamin C and selenium on semen quality and sperm parameters, such as motility, viability, morphology, and concentration, against the adverse effect of stress.

    Keywords: Sperm, Infertility, ascorbic acid, Selenium, Stress
  • B. I. Abilov *, K. B .Isbekov, S. Z. Assylbekova, N. B. Bulavina, G. A. Kulmanova, Koishybayeva S. K, L. Nikolova Pages 1143-1154

    The objective of changing the simple exploitation of fish stocks to highly efficient fish farms in lakes and reservoirs is to improve the productivity of inland freshwater fish. The small- and medium-sized lakes can be used to increase the production of farmed fish with lake management. Therefore, this study proposed to investigate the production and economic efficiency of carp in lake commercial fish farms. In this investigation, the results of carp farming experiments in fish farms in small lake commercial fish farms (LCFF) are evaluated using advanced methods and techniques. The research was carried out based on the Voroshilovsky reservoir, which operated in the LCFF mode. The farm had hatchery and carp fry ponds for expanding fingerlings based on the "Scientific and production center of fishery" LLP from 2019 to 2020. This study was performed on different types of common carp and herbivorous fish (grass and silver carp), and sexual products were collected in fried ponds and the Voroshilovsky reservoir. The absolute growth gain of common carp, silver carp, and grass carp were 301.00, 300.40, and 577.00 grams, respectively, and their mean daily weight gain values were 2.50, 2.50, and 4.80 grams. Common carp recorded the highest level of planned fish productivity (169.30 kg/ha), and the lowest level of this trait was grass carp (43.50 kg/ha). Data of mean weight and body length of common carp, grass carp, and silver carp fishes showed a variation of 4.55 kg and 56.25 cm, 6.06 kg and 75.50 cm, as well as 6.30 Kg and 75.05 cm, respectively. This difference can be justified according to the variance of fish length, which on average, 80 grams of weight is obtained per centimeter of fish length. The economic efficiency of carp was calculated, and the net profit was determined at more than 50% of total income. According to the net profit indicator, due to the implementation of a part of the fingerlings, the carp reared in the pond area; as a result, this method is profitable and accounts for 104% of the total planned economic profit. Therefore, fish production from aquaculture can rise to 10 times to maintain high-quality food security and other essential nutrients, provide job opportunities, and cash income to help job-seeking youth.

    Keywords: Aquaculture, Commercial fish, CARP, Lake-commercial fish farm, Reclamation, Reservoir, Stocking material