Development of a Novel Oral Vaccine Against Human Resipratory Syncytial Virus

Human respiratory syncytial virus (HRSV) is the leading cause of bronchiolitis and pneumonia in infants, children, the elderly and the immune-compromised. The goal of immunization is to provide sufficient protection to prevent serious lower respiratory tract diseases leading to hospitalization and reducing the frequency of complications such as otitis media. Prevention and treatment of HRSV infection using antiviral agents is challenging because it is a rapid acute infection and by the time the infection is recognized it may be too late to control the disease with any antiviral therapy alone. Thus, there is a worldwide need for an HRSV vaccine. Studies have shown that the immunogenic domains of F and G proteins could confer protection against HRSV infection in vaccinated hosts. In the present study, firstly the immunogenic domain of HRSV G domain was expressed in Escherichia coli . Then a rabbit was immunized using purified-recombinant G domain protein. The results of neutralization assay showed that G domain alone could raise active polyclonal antibodies against HRSV successfully. Secondly, the potential of G and F immunogenic domains as vaccine candidates were studied by using live bacterial vaccines. Both the G and F domains were separately initially cloned in pKMSInak plasmid before they were surface displayed on Salmonella typhi Ty21a used as the delivery system. The surface displayed G and F domains were detected using indirect immunofluorescence, sero-agglutination and outer membrane protein separation approaches suggesting that the Inak protein successfully carried the G and F domains to the surface of Salmonella cells. For in vivo evaluation of the designed vaccines, Balb/c mice were immunized orally with live Salmonella cells harboring pKMSInak-G or pKMSInak-F and challenged against HRSV. The humoral (TH2), cellular (TH1) and mucosal immune (IgA) responses of the immunized mice were studied by measuring cytokines (IL-2, IL-4, IL-5, IL-9, IL-10, IL-12, IL-13, IL-17, IFN-γ and TNF-α), chemokines (RANTES and MIP-α) and immunoglobulins (IgG, IgG1, IgG2a, IgG2b and IgA) levels in their sera before and after challenging with HRSV. Lymphocyte proliferation assay was performed to evaluate the cell mediated immunity. Histopathological examinations were also carried out as confirmatory tests. The results showed that pKMSInak-G and pKMSInak-F vaccines could significantly enhance TH1 and TH2 responses as well as mucosal immunity in the immunized mice compared to the control group. Histopathological examinations indicated that the immunized mice had significantly lesser lung tissue damage than the control. Moreover, the obtained ratios of TH1/TH2 were desirable (~1) suggesting that Salmonella cells carrying pKMSInak-G and pKMSInak-F are potent vaccine candidates against HRSV

Isolation and optimization of Asparaginase producing bacteria in solid state fermentation by Plackett-Burman desighn

زمینه و هدف: این تحقیق به روش تخمیر در بستر جامد به منظور بررسی انواع بسترهای جامد در تولید آنزیم آسپارژیناز توسط سویه ی جداسازی شده از منطقه لاکان استان گیلان از جنس باسیلوس انجام گردید. روش بررسی: در این بررسی تجربی جهت جداسازی باکتری های تولید کننده ال آسپارژیناز از محیط کشت M9 استفاده شد. جهت غربالگری در بستر جامد از بسترهای مختلف سبوس برنج، ساقه برنج، سبوس گندم و ساقه گندم به عنوان منابع کربن و همچنین از عصاره مخمر، پپتون، آسپارژین، تریپتون به عنوان منابع نیتروژن استفاده شد. جهت بهینه سازی محیط کشت و بررسی اثر منابع کربن و نیتروژن بر میزان تولید آنزیم آسپارژیناز با استفاده از روش Plackett-Burman تعداد 12 آزمایش در 2 سطح طراحی شد؛ سپس انواع بسترهای جامد با تغییر فاکتورهای تأثیرگذار در تولید آنزیم مورد ارزیابی قرار گرفت. یافته ها: بهترین بستر جامد، بستر سبوس برنج و بهترین منبع ازت آسپارژین، بیشترین تأثیر را در افزایش تولید آنزیم از خود نشان دادند. بهترین شرایط تولید آنزیم به دست آمده در بستر جامد در میزان غلظت سبوس برنج 5/1 وزنی/ وزنی و میزان غلظت آسپاراژین 5/0 وزنی/ وزنی و دمای 30 درجه سانتی گراد و به مدت زمان 72 ساعت به میزان 24/69 واحد آنزیم بر گرم مشاهده گردید. نتیجه گیری: در این تحقیق میزان تولید آنزیم با استفاده از سبوس برنج به عنوان منبع کربن در بستر جامد پس از بهینه سازی نسبت به شرایط بهینه سازی نشده افزایش قابل ملاحظه داش

Proteins expression clustering of Alzheimer disease in rat hippocampus proteome

Because of the huge amounts of proteomic data and demand for new methods of laboratory analysis results, proteins collective analysis, in addition to taking less time, biostatistician assist at identification of new patterns in the data set. In this study, rat hippocampus proteome in normal and Alzheimer's disease (AD) were analyzed by using proteomic techniques and bioinformatics’ analysis. Protein extracts from normal and Alzheimer's rats were separated by using two-dimensional electrophoresis (2DE). The silver staining method was used for detecting spots. Bioinformatics analysis of proteome were performed by progensis same spots software. Bioinformatics and statistical analysis of 2DE gel techniques obtained 760 protein spots were detected in both normal and AD rats.  Comparisons between controls and Alzheimer gel containing 20 common proteins were expressed significantly differences. 16 new proteins were expressed in AD, while 36 proteins were suppressed. Proteins clustering by using correlation analysis evaluated 3 clusters in the proteome; Principal component analysis also confirmed the results of clustering. Finally, we can conclude that a significant expression of Alzheimer changes in the hippocampus proteome which are associated with specific biological processes summarized in 3 main clusters indicated 3 principal biological pathways of AD.

How to test normality distribution for a variable: a real example and a simulation study

Many commonly used statistical methods require that the population distribution be nearly normal. Unfortunately, in some papers the one-sample Kolmogorov-Smirnov test has been used for testing normality while the assumptions of applying this test are not satisfied. To conduct this test, it is assumed that the population distribution is fully specified. In practical situation where the mean and SD of population distribution is not specified in advance, one can use a modification of the K-S test for checking the normality assumption which is called, Lilliefors test. In this paper, we explain the method of computing this test with some common statistical softwares such as SPSS, S-PLUS, R and StatXact and utilize a dermatology dataset from Skin Research Center of Shohada-e-Tajrish hospital to illustrate how the use of the one-sample K-S (with the mean and SD estimated from the sample) instead of its modification can be misleading in practice. We also use Monte Carlo simulation to compare the approximate power of the one-sample K-S test (with the estimated population mean and SD) with Lilliefors test in some common specified continuous distributions. The result indicates that one should not use the one-sample K-S test for assessing the normality assumption in practical situation.

Biomarker Profiling (Erbb2, P53, and PR) for Stage I of Breast Cancer

Breast cancer (BC) accounts for one of the major health problems around the world. Since the diagnosis process can have great effect on therapy outcomes, we studied the biomarkers specific to breast tumors stage I based on examining different Iranian patients. Cases from different stages were examined to discover their highly expressed proteins. In addition, pathologic evaluations were performed as the diagnosis procedure. Considering positive percentage of over-expressed protein in different stages in the population, it is guessed that over-expression of ErBb2 and PR are positively correlated, while P53 is in negative correlation with them. Therefore, these molecules can probably account for stage I biological marker. This study suggests that alterations in over-expression of specific biomarkers in different stages may be associated to the stage classification, and can help achieve more effective therapies of this malignancy

Positive Result for SARS-CoV-2 RNA Test after a Long Time for the Patient with COVID-19 even after Discharge from the Hospital

Background: Ruthin's coronavirus disease 2019 (COVID-19) diagnosis is based on the positive result of real-time polymerase chain reaction (PCR) from the nasal and oropharyngeal swab. However, chest CT scans can play an important role in diagnosing patients with COVID-19. Cases Report: In this study, we reported a 44 years old female with a mild form of the COVID-19 who showed a positive result for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA for 44 days after symptom onset. The suspected case was detected using real-time PCR. After two weeks of hospitalization, the patient was discharged, but her molecular tests were performed twice after one month and 44 days, and they remained positive for SARS-CoV-2 RNA. Conclusion: In theory, if the patient becomes re-infected or the virus reacts, these individuals may serve as a transmission source. So far, the only way to screen for possible reinfection has been by using PCR on separate specimens

Antibacterial effects of Scrophularia striata seed aqueous extract on staphilococcus aureus

Plant-based drugs are regarded promising recently. Scrophularia has been shown various biological activities such as antimicrobial, antitumor and anti-inflammatory properties. According to pervious researches, bacteria are becoming resistant to some kinds of antibiotics, so it is prominent to find more reliable sources against them. Here Staphylococcus aureus as a common type of Gram- positive bacteria has been chosen for this in vitro study. After culturing this bacterium, it was treated with various dosages of Scrophularia striata seed aqueous extract and tetracycline, and then its antibacterial effect was assessed by spectrophotometery method based on bacteria population alteration after 24h incubation. In order to investigate probable side effects of the seed extract its effect on human fibroblast cells has been studied. MTT assay was applied for cell survival determination of human fibroblast cells after 24h. Findings indicate that bacterial population has been declined between 1 to 20 μg mL-1 dosages of the extract, which 5 μg mL-1 is the most effective dosage without containing any cytotoxic effect on human fibroblast cells. In fact extract not only has no antiproliferation properties but also has evoked cell profileration, so it can be consider as a cell growth factor. Tetracycline, on the contrary, showed its potent impact merely in highest dosages with noticeable side effects. It can be concluded that, Scrophularia striata could be possibly a promising antibiotic which shows significant antibacterial properties, and with lesser side effects.

Survey of protection role of selenium in reduction of T-2 toxin effect on phagocytosis function of human neutrophil

زمینه و هدف:مایکوتوکسین T-2 اثرات جدی بر سلول های سیستم ایمنی می گذارد. این مطالعه با هدف بررسی اثر سم T-2 بر روی فعالیت فاگوسیتوزی نوتروفیل خون محیطی انسان و نقش حفاظتی سلنیم بر این فرآیند طراحی و اجرا شده است. روش بررسی:در این مطالعه تجربی، نوتروفیل خون محیطی انسان با استفاده از روش دکستران- فایکول خالص سازی گردید. برای تعیین میزان فعالیت فاگوسیتوزی نوتروفیل ها از روش شمارش تعداد بلع مخمرهای کاندیدا آلبیکانس اپسونیزه استفاده شد. سپس میزان فعالیت فاگوسیتوزی نوتروفیل در حضور غلظت های مختلف سم T-2 مورد بررسی قرار گرفت. در نهایت، اثر غلظت های مختلف سلنیم در کاهش اثرات سم T-2مطالعه شد. از روش آماری آنالیز واریانس در نرم افزارStata جهت تجزیه و تحلیل داده ها استفاده شد. یافته ها:سم T-2 در غلظت µg/ml 100بر فعالیت کاندیدا کشی نوتروفیل تأثیر گذاشته و این فعالیت را تقریباً به نصف کاهشداد. سلنیم در غلظت µg/ml100 بهترین اثر را داشت و فعالیت کاندیداکشی را نزدیک به 90 درصد بهبود بخشید. نتیجه گیری:براساس نتایج این مطالعه سم T-2 موجب کاهش شدید فعالیت فاگوسیتوزی نوتروفیل ها می گردد و سلنیم اثر حفاظت بخش دارد و می تواند فعالیت فاگوسیتوزی را تقریبا به حالت طبیعی بازگردان

Evaluation and comparison of immunization level between recombinant proteins of binding subunit of entrotoxigenic Escherichia coli and botulinum toxin

زمینه و هدف: در میان عوامل باکتریایی، شایع ترین عامل بیماری اسهال، باکتری اشریشیاکلی انتروتوکسیژنیک است. زیر واحد LTB سم این باکتری قادر به ایجاد مصونیتی شش ماهه است. کلستریدیوم بوتولینوم نیز عامل بیماری کشنده بوتولیسم می باشد و زیر واحد BoNT/A-Hc سم آن می تواند تا دو سال در برابر این بیماری مصونیت ایجاد کند. میزان ایمنی زایی که هر یک از این زیر واحدهای نوترکیب ایجاد می کنند، می تواند از عواملی باشد که در به وجود آمدن مصونیتی با ماندگاری متفاوت تأثیرگذار باشد. هدف از این مطالعه بررسی و مقایسه میزان تولید آنتی بادی ناشی از استفاده از دو پروتئین LTB و BONT/A-Hc در موش آزمایشگاهی بود. روش بررسی: در این مطالعه تجربی از باکتری Bl21DE3 E. Coli تراریخت شده با وکتور pET28aاستفاده گردید. این وکتور حاوی ژن نوترکیب LTB اشریشیاکلی انتروتوکسیژنیک و ژن نوترکیب BONT/A-Hc بوتولینوم به طور جداگانه بود. پس از بهینه سازی بیان و تخلیص پروتئین نوترکیب LTB از فاز نامحلول و BoNT/A-Hc از فاز محلول عصاره سلولی، محصولات آن ها بر روی ژلSDS-PAGE بررسی گردید. موش های آزمایشگاهی به وسیله پروتئین های حاصل، ایمنی زایی شدند. تیتر آنتی بادی حاصل از هر دو نوع پروتئین نوترکیب با استفاده از آزمون آماری t-Test در نرم افزار SPSS ارزیابی و مورد مقایسه قرار گرفت. یافته ها: پروتئین های نوترکیب بیان شده BoNT/A-Hc و LTB با ستون نیکل تخلیص شدند. پس از ایمنی زایی موش های آزمایشگاهی، تفاوت معناداری در تیتر آنتی بادی برای دو پروتئین BoNT/A-Hc و LTB مشاهده گردید (01/0>P). نتیجه گیری: اختلاف کمی بین تیتر آنتی بادی برای دو پروتئین BoNT/A-Hc و LTB مشاهده گردید که می تواند به دلیل خاصیت قوی ادجوانسیتی LTBو ایمنی زایی ایجاد شده تقریباً یکسان، در فاصله زمانی محدود باش

Autophagy Role as a Double-Edged Sword in Anesthesiology and Critical Care

Autophagy is a mechanism, which host cells can utilize it to defend against infections. Trapped cargo such as viral cargo and delivered to a lysosome for degradation. Primary immune response against some viruses can start by autophagy mechanism. In this study, we reviewed role of autophagy in viral infections