zahra noormohammadi

Endometriosis is a complex disease that can be hereditary and its occurrence is under many genetic and environmental factors. Genome-wide studies have proven that they can be successful in identifying variants that affect the occurrence of complex diseases. In the present study, the analysis and summation of genome-wide studies in relation to WNT4 and CDKN2B-AS1 gene variants, and their association with the occurrence of this disease, has been done.

In the present meta-analysis, studies on polymorphisms associated with WNT4 and CDKN2B-AS1 genes in the GWAS database have been reviewed. The studies were related to populations with European and East Asian ancestors, and in order to investigate the variant with the highest level of association in the above gene loci with the occurrence of endometriosis, the fixed-effect statistical model was used.

Among the examined variants, loci rs3920498 (with significance p<1.1e-5 and probability ratio equal to 1.19) and rs1537377 (with significance p<1.33e-10 and probability ratio equal to 1.09) are more associated with endometriosis.

In the present study, it was found that rs3920498 (upstream of WNT4 gene) and rs1537377 (downstream of CDKN2B-AS1 gene) loci are most associated with endometriosis disease.

Biofilms are communities of microorganisms which are difficult and complicated to treat with antimicrobial agents. Therefore, there is a need to use new compounds to treat these types of resistant microorganisms. This study was done with the aim of finding an effective compound for inhibition of biofilms of Pseudomonas aeruginosa, Escherichia coli, Streptococcus mutans, and Staphylococcus aureus.

The effect of bacteriocin-like compounds produced by B. subtilis SP1 isolated from Sablan native honey on the biofilm of four bacterial strains including E. coli, P. aeruginosa, St. mutans and S. aureus was investigated by using 96-well microtiter plate and crystal violet staining.

The percentage of anti-biofilm effect against S. mutans and S. aureus, E. coli and P. aeruginosa at 80 µg/ml of bacteriocin like compounds were about 80, 62, 50, and 15 percent, respectively. The antibiofilm effect was concentration dependent and reduced at lower concentrations of bacteriocin like compounds.

The highest percentage of anti-biofilm effect of bacteriocin like compounds was observed against S. mutans and the lowest percentage of inhibition was against P. aeruginosa.

This study aimed to investigate the relationship between follicular fluid Bisphenol A (BPA) concentrationswith alterations in the expressions of NOTCH1-3, CASPASE 3/7, HLA-G, and ICAM-1 genes and the number ofretrieved mature oocytes (MII oocyte) in the cumulus cells of infertile poor ovarian response stimulates women.In this prospective cohort study, 80 infertile unexpected poor ovarian response (POR) subjectswere selected on the basis of subgroup 1a of the POSEIDON classification. They were divided into two groups: group1 consisted of 40 women, each with a higher number of metaphase II (MII) oocytes (G1, 3-4 oocytes retrieved), whilegroup 2 comprised of 40 women, each with a lower number of MII oocytes (G2, ≤2 oocytes retrieved). The expressionsof the studied genes were evaluated by quantitative-real time polymerase chain reaction (PCR). The concentration ofBPA in follicular fluid was measured with HPLC. The expression levels of NOTCH1-3, HLA-G, and ICAM-1 genes were significantly lower in G2 than G1(P<0.05). Meanwhile, CASPASE 3/7 expression levels were higher in unexpected POR patients in G2 compared toG1 (P<0.05). There was a significant direct correlation between the levels of NOTCH1-3, HLA-G and ICAM-1 geneexpressions and there was also a significant inverse correlation (P<0.05) between the levels of CASPASE 3/7, with thenumber of MII oocytes and embryo development between the two groups. The concentration of BPA in the follicularfluids of G2 was higher compared to G1 (P<0.05). A higher concentration of BPA was associated with a lower number of mature oocytes and oocyte qualityin these patients. Also, alterations of NOTCH1-3, CASPASE 3/7, HLA-G, and ICAM-1 transcript levels in unexpectedPOR women were associated with BPA concentration.

The chimer structure consisting of the extracellular domain of CTLA4 and the FC fragment of human IgG1 antibody binds to CD80 (B7-1) and CD86 (B7-2) ligands on APCs as a result inhibits CD28 binding and T lymphocyte activation. This structure, called Abatasept, was approved by the US FDA in 2005 to treat rheumatoid arthritis.

In this study, the CTLA4-FC chimer structure was designed and then some of its features were investigated using bioinformatics tools. The gene was then cloned into the pET-28a plasmid and transformed into E.coli BL21DE3. Recombinant protein was expressed and purified, and flow cytometry was used to evaluate the binding affinity. Mixed lymphocyte culture (MLR) method was also used to evaluate the biological activity of recombinant protein.

Bioinformatics results showed that the designed structure of CTLA4-FC is suitable in terms of physicochemical properties, second and third structure, how it binds to the receptor, antigenic properties and mRNA properties. Recombinant protein was well expressed in bacteria. Purification was performed as required. Western blotting and ELISA results were positive and flow cytometry and MLR tests were acceptable.

Recombinant CTLA4-FC protein was produced in bacterial cells and bound well to CD80 and CD86 receptors and was able to inhibit T cells. So it can be a good candidate for drug production.

Hearing loss is the second most common disease after mental retardation in Iran.Autosomal recessive non-syndromic hearing loss (ARNSHL) is an extreme and highly heterogeneous disease, for which more than 70 genes have been identified. Considering the frequency of family marriage as well as the im-portance of ARNSHLin Iran, we evaluated the genetic factors involved in this type of deafness.

We performed the whole exome sequencing (WES) of eight Iranian subjects with severe non-syndromic hearing loss selected from 110 well-characterized subjects with non-syndromic hearing loss from 2017-2019.The patients with mutated GJB2and GJB6genes were excluded from the study.

The use of the whole exome sequencing method revealed 10 different mutations in 7 genes, including SLC26A4 (c.1234G>T), FGF3(c.45DelC, c.466T>C), ADGRV1(c.12528-2A>C, c.16226-16227insAGTC), OTOG(c.7454delG), OTOF(c.3570+2T>C), ESPN(c.992G>A), OTOA(c.2359G>T, c.2353A>C). Seven new variants were observed in seven families including SLC26A4 (c.1234G>T), FGF3(c.45DelC), ADGRV1(c.12528-2A>C), OTOG(c.7454delG), ADGRV1(c.16226-16227insAGTC), OTOF(c.3570+2T>C).

The causal mutation of ARNSHLwas found in all patients using the WES. Meta-analysis studies can help to identify common mutations causing deafness in any population to facilitate identification of carriers and subjects with deafness.

Von Willebrand disease (VWD) is the most common inherited bleeding disorder around the world. Large size of the gene and heterogeneous nature of its mutations, make it difficult to determine the exact nature of mutations in VWD. Some missense mutations are common in VWD and can be assessed as first step of determining the genetic cause of the disease.

Forty four unrelated patients categorized as VWD type 3 were included in study. Tetra ARMS-PCR was applied to determine the presence of the point mutations rs61753984 and rs62643623. Then ARMS-PCR was performed with the same primers. In cases with mutant patterns, Sanger sequencing was used for conformation of results.

Two patients showed two bands of 248bp and 181bp in rs61753984 (R34X), which corresponds to homozygous mutant genotype. In other patients, homozygous pattern with 248bp and121bp bands was observed. In rs62643623 (Q218X) position, one patient with two bands of 378bp and 260bp showed homozygous mutant genotype while other patients showed homozygous wild genotype (378bp and 168bp bands). The results of ARMS-PCR was the same as tetra ARMS-PCR. Finally, the results of tetra ARMS-PCR were confirmed by Sanger sequencing.

In present study we used tetra ARMS-PCR for detection of point mutations, while in most studies PCR-PFLP has been used. We found that tetra ARMS-PCR can be used as a simple, cost effective and reliable method. If these mutations are not found, advanced molecular methods such as sequencing should be applied.

 Cancer is mainly caused by defect in natural regulation of programmed cell death (PCD). On the other hand, it is proved that, the EGFR is overexpressed on the surface of many malignant cell. Therefore, immunotoxin against EGFR, which consisting an antibody or fragment of an antibody linked to the toxic moiety is one of the most reliable strategies for cancer treatment. Here we evaluate the effect of a novel immunotoxin molecule harboring ricin, as a toxic moiety on the mRNA level of two main apoptotic molecules on cancerous cell lines.

 The Effects of immunotoxin molecule and it’s components including ricin toxin and antibody fragment on the mRNA level of pro- apoptotic BAX and anti-apoptotic BCL-2 and EGF receptor genes on the breast MDA-MB-468 and colorectal HCT-116 cancer cell lines were assayed by qRT-PCR. We used glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as a reference gene.

 Treatment with immunotoxin significantly increased the expression of BAX gene and decreased the expression of EGFR and BCL-2 genes (P<0.05) and induced apoptosis. Also, ricin toxin caused a significant decrease in BCL-2 and no significant increase in BAX in cancer cells and decreased EGFR only in breast cancer cells. The scFv part had no significant changes on genes expression.

 The present findings showed that the recombinant Panitumumab-Ricin protein, may be an appropriate candidate for the treatment of EGFR overexpressed cancers by induction in apoptosis pathway.

Vitis vinifera L. is one of the most important plant products in the world .The aim of this study was to investigate the anatomical structure of leaves and petioles of seven grape cultivars of Boroujerd region with light microscopy. Different anatomical features such as leaf thickness, petiole diameter, diameter of vein and petiole vessels, diameter of leaf and petiole bundle, thickness of leaf and petiole epidermis, diameter of main vein, number of petiole and main vein clusters and diameter of brain parenchyma. According to the results, the highest leaf thickness, petiole diameter, diameter of vein and petiole vessels, diameter of leaf and petiole vascular bundle, diameter of petiole nucleus parenchyma and number of petiole vascular bundles were related to ruby cultivar. In the case of traits such as leaf thickness, leaf epidermal thickness, diameter of vein vascular bundles, diameter of leaf vascular bundle, main vascular bundles, the lowest value was shown by black grape cultivar. Manqa cultivar showed the highest leaf epidermis thickness and main vein diameter. The smallest diameter of the main vein, the diameter of a petiole bundle and the thickness of the petiole epidermis belonged to Fakhri cultivar. Asgari cultivar showed the highest value of main vein vascular apparatus and the lowest value of petiole diameter, diameter of petiole vascular bundles and diameter of petiole brain parenchyma. Sahebi cultivar showed the highest thickness of petiole epidermis. Comparison of anatomical structure of leaves and petioles showed significant differences in the studied indices in different cultivars.

Two newly identified proteins, EspB and EspC are involved in the pathogenesis of Mycobacterium tuberculosis. The objective of the present study was to evaluate the immunogenicity of recombinant EspC, EspB, and EspC/EspB fusion proteins in mice.

BALB/c mice were immunized subcutaneously with recombinant EspC, EspB, and fusion EspC/EspB proteins, three times with along with Quil-A as an adjuvant. The cellular and humoral immune responses were evaluated by quantifying IFN-g, IL-4, IgG, IgG1, and IgG2a antibodies against the antigens.

The results showed that the mice immunized with recombinant EspC, EspB, and EspC/EspB proteins did not produce IL-4, whereas IFN-g was secreted in response to all three proteins. EspC/EspB group produced significant amounts of IFN-g in response to stimulation with all the three recombinant proteins (P<0.001). In mice immunized with EspC, high levels of IFN-g were detected in response to EspC/EspB, and EspC (P<0.0001); while mice immunized with EspB produced lower levels of IFN- g in response to EspC/EspB, and EspB (P<0.05).

All the three recombinant proteins induced Th1-type immune responses in mice against EspB and EspC; however, EspC/EspB protein is more desirable due to the presence of epitopes from both EspC and EspB proteins and the production of immune responses against both.

Rhamnolipids are one of the most important bio surfactants of glycolipids produced by Pseudomonas aeruginosa. In this study, the expression of bio surfactant producing proteins of Pseudomonas aeruginosa in the presence of sodium dodecyl sulfate coated iron nanostructure (Fe/SDS) were evaluated.

The expression of Surfactant Protein A enzyme was investigated by SDS-PAGE and Western blot. The enzyme rhamnosyl transferase was analyzed on SDS-PAGE gel and confirmed by Western blotting. The presence of nanoparticles on the surface of bacteria and the morphology of nanoparticles were investigated by TEM and SEM.

According to SEM, nanoparticles with a size of 20 nm and a spherical shape were seen. Binding of Fe / SDS nanoparticles to bio surfactant was confirmed by TEM. The enzyme rhamnosyl transferase with a molecular weight of 40 kDa was expressed in the test sample. The results of protein expression showed that the confirmed of rhamnosyl transferase enzyme increased significantly under the influence of Fe/SDS nanoparticles and showed the confirmed compared to other nanoparticles and the control sample. While SDS and Fe nanoparticles alone are not effective in confirmed this protein and are almost identical to the control sample. Western blotting results of SDS-PAGE confirmed the expression of rhamnolipid transferase under the influence of Fe/SDS nanoparticles.

The results of the present study indicate the importance of Fe/SDS nanoparticles in increasing the expression of rhamnolipid transferase enzyme in the production of bio surfactants from Pseudomonas aeruginosa.

 Dysregulation of long-term expression of non-coding RNAs (lncRNAs) has a potential role in progressive brain disorders such as Alzheimer's disease. This study aimed to analyze the apoptosis and expression of 51A and NAT-Rad18 lncRNAs and their target genes in brain tissue and peripheral blood mononuclear cells (PBMCs) of the rat model of AD, before and after memantine treatment.

 Twenty-eight male Wistar rats were divided into four groups: 1. Normal control (n = 4), 2. Sham-operated (n = 4), 3. Alzheimer's control (n = 10), and 4. The experimental group (n = 10) was treated with memantine. The qPCR and TUNEL tests were used to detect the lncRNAs expression and apoptosis.

 Sorl1 gene was reduced in brain tissue of Alzheimer’s control (p = 0.016) and PBMCs of Alzheimer's control and experimental groups (p = 0.002 and p = 0.001 respectively). The expression of NAT-Rad18 and Rad18 genes increased in brain tissue of Alzheimer's control group (p = 0.002 and p = 0.04 respectively) while reduced in PBMCs of Alzheimer's control and experimental groups (p = 0.005 and p = 0.045 for NAT-Rad18, p = 0.01 and p = 0.006 for Rad18).

 ROC curve analysis showed 100% sensitivity and 85.7% specificity for the Sorl1 gene with 0.911 under the curve area and 100% sensitivity and 100% specificity for NAT-Rad18 and Rad18, separately with one under the curve area. Decreased expression in Sorl1, NAT-Rad18, and Rad18 genes can be used as blood biomarkers for diagnosis independently. However, studies on Alzheimer's patients are needed.

Multi-drug resistant (MDR) Klebsiella pneumoniae strains cause the majority of community acquired and life-threatening infections. We aimed to detect the gyrA mutations in the clinical strains of nalidixic acid and ciprofloxacin resistant K. pneumoniae isolated from the patients with urinary tract infections.

Bacterial strains were isolated from the patients with urinary tract infections admitted to a major hospital in Tehran, Iran (2017-2018). Bacterial identification was done according to standard microbiological tests. Antimicrobial susceptibility testing for quinolones and fluoroquinolone antibiotics was done using both disc diffusion and minimal inhibitory concentrations (MICs) methods. PCR-RFLP was used to detect the probable mutation in the gyrA gene in nalidixic acid and ciprofloxacin resistant strains. Finally sequencing was performed to detect point mutations in isolated K. pneumoniae strains.

One hundred K. pneumonia isolates were recovered from the urine samples of the clinical cases. Antibiotic resistance testing showed that among all K. pneumoniae isolates, 26% and 19% of the strains were resistant to nalidixic acid and ciprofloxacin respectively. MIC value was ≥4 µg/ml for ciprofloxacin resistant isolates. The results of RFLP on gyrA PCR amplicons using HinfI restriction enzyme showed point mutation in this gene in 46% of nalidixic acid and ciprofloxacin resistant K. pneumonia. The data obtained from the sequencing confirmed the RFLP results and indicated the presence of point mutations in codons 83 and 87 in the gyrA gene which leads to the substitution of different amino acids in gyrA protein.

Our findings indicated a relative increased rate of resistance against quinolones and fluoroquinolone antibiotics that raised a concern about extensive dissemination of clinical strains of nalidixic acid and ciprofloxacin resistant K. pneumonia. Point mutation of gyrA gene was responsible for the resistance in our strains however to gain more insight into the molecular characterization of quinolone-resistant isolates, other possible mechanisms of the resistance should also be investigated.

Loneliness is one of the most common problems of elderly aging, which greatly affects the health of the elderly. Lifestyle is one of the most important factors affecting health. This study was conducted to explore the relationship between loneliness and healthy lifestyle in the elderly 60-75 years in Gonabad city in 2017.

In this cross-sectional study, 263 elderly people in Gonabad were selected by stratified random sampling. In order to collect information, a questionnaire of demographic information, feeling of loneliness and healthy lifestyle in the elderly was used. Descriptive and inferential statistical tests (independent t-test, analysis of variance and Spearman correlation coefficient) and SPSS version 20 were used to analyze the data. Significance level less than 0.05 was considered.

The mean score of loneliness and stuttering style of elderly was 65.26±11.65 and 153.19±27.95. Furthermore, a significant and reverse relationship was detected between loneliness and healthy lifestyle in the elderly. Thus, lifestyle score was higher in the elderly who had less loneliness (P=0.00, R=-0.72).

The results of this study show that reducing loneliness  has an effective role in improving lifestyle of elderly. With this in mind, the role of supporting nursing caregivers, educating specialist staff in the field of egalitarian medicine, family counseling and mental health for the elderly and their families are highlighted.

Pelargonium graveolens L ′Her is an aromatic and medicinal plant which is widely used in pharmaceutical, perfumery and food industry. In this study, the effect of ultraviolet radiation with different intensities (0, 0.12, 0.26 and 0.38 W/m2) was investigated on P. graveolens in InVitro condition. The amount of total phenol, flavonoid and anthocyanin, activity of phenylalanine ammonia lyase (PAL), photosynthetic pigments, carbohydrates, proteins and activity of some anti-oxidant enzymes (Catalase, peroxidase and superoxide dismutase) were evaluated. The antioxidant activity of methanolic extract was measured by using 2-2-di-phenyl-1-picaril hydrazil (DPPH) method. Total protein content and chlorophylls a, b showed a significant decrease (P≤ 0.05), while the amount of carotenoid increased significantly (P≤ 0.05), when ultraviolet radiation increased. With increasing UV-B radiation, the activity of studied enzymes, total phenols, flavonoids and anthocyanins content increased significantly (P≤ 0.05). Also, with increasing radiation intensity, the mean IC50 of methanolic extract decreased. The decreasing pattern of IC50 was consistent with the trend of increasing the mean of phenols. The present results show that P.graveolens in response to UVB stress has increased its antioxidant capacity by increasing the activity of antioxidant enzymes and increasing the phenols.

 Klebsiella pneumoniae is one of the three pathogens that has become a global disease control and treatment problem due to its resistance to common antibiotics. For this reason, it is crucial to study the genes that cause antibiotic resistance in it. Therefore, the aim of this study was to investigate the phenotypic and genotypic frequency of tetracycline resistance in clinical isolates of K. pneumoniae in Tehran, Iran.

 In this study, 100 isolates of K. pneumoniae isolated from clinical samples (urine) during 2018-2019 were studied. In addition to microbial and biochemical phenotypic tests, genotypic tests were conducted to determine the frequency of antibiotic resistance genes tet (A, B, C, 39).

 Out of 100 isolates of K. pneumoniae, 49 isolates were resistant to tetracyclines. The results of multiplex PCR showed that 31 samples were positive for tetA gene, 8 isolates for tetB gene, 21 samples for tetC and, and 8 isolates for tet39. None of the isolates were positive for all four tetracycline genes.

 The results of this study showed that the isolates were positive for at least one gene and at most 2 tetracycline resistance genes. The tetA gene showed the highest frequency and the lowest frequency was demonstrated by tetB. The highest binary combination of genes was tetA-tetC, and the lowest was tetA-tet39.

Rabies is almost always fatal but entirely preventable through proper vaccination. Inadequacy of costly high-quality cell culture vaccines is sometimes a bottleneck for expanded rabies control plans. Reverse genetics along with other molecular biology means are trying to improve the immunogenicity and yield of rabies vaccine products.

An additional glycoprotein gene of the rabies virus PV strain was inserted between the glycoprotein and polymerase genes of the virus. The viral proteins were expressed at the T7BHK cell line to rescue the recombinant virus.

The recombinant virus containing two consecutive glycoprotein genes was rescued from T7BHK cells. The virus particles were functional and successfully infected the permissive BSR cell line.

The new virus strain with an additive copy of the glycoprotein gene has a good potential to be utilized in different studies, including cell biology and immunological properties of the rabies virus. In this study, the recombinant rabies virus was successfully rescued from cell culture which would pave the way for further investigations on this virus.

Soil microorganisms play an important role in the biological control of pathogenic fungi. The aim of this investigation was to isolate soil-borne Bacillus subtilis with the ability to produce antifngal lipopeptides  that are suitable  for suppressing Fusarium graminearum, which contaminates wheat, corn, potato and a wide range of plants.Materials &

B. subtilis was isolated from the rhizospher of healthy plants of park slocated in the five areas north, south, east, west and center of Tehran and was identified based on morphological characteristics, biochemical tests and sequencing of 16S rRNA. The antagonistic activity of isolated strains against F. graminearum was investigated by Well method. The strains that inhibited the growth of fungi and showed the greatest inhibition zone, were selected. Surfactin of selected bacteria was purified by methanol method and bacterial metabolites and pure surfactin (Sigma Company) were compared with, high performance liquid chromatography (HPLC).

Among 60 isolated strains, 27 strains had antifungal activity. Six strains with the highest fungal inhibition zone (8-16 mm) were selected and their yellow color and transparency was a confirmation to HPLC. Two bacteria with the highest amount of surfactin production by  molecular method showed high similarity with B. subtilis.

The results show that Bacillus subtilis isolated from soil are good candidates for biological control of plant pathogenic fungi and therefore can a suitable alternative to chemical fungicides.

SLC39A6 (solute carrier family 39) or LIV-1, is a zinc-transporter protein associated with estrogen-positive breast cancer and its metastatic spread. Significantly there is a direct relation between high zinc intake and unregulated proliferation and cancers. Blocking SLC39A6 protein may result in reduced metastasis and proliferation in many malignant tumors. This study aimed to develop an anti-SLC39A6 nanobody that is able to detect and block the SLC39A6 protein on the surface of cancerous cells.

The recombinant SLC39A6 was expressed and used for camel immunization. The VHH library was constructed and screened for SLC39A6-specific nanobody. Then, the strength of nanobody in SLC39A6 detection was evaluated by Western blotting and flow cytometry.

We showed the ability of nanobody to detect SLC39A6 by Western blotting and flow cytometry and the specificity of the C3 nanobody for the SLC39A6 antigen. Furthermore, the selected nanobody potently inhibits cell proliferation.

These data show the potential of SLC39A6-specific nanobody for the blockade of zinc transportation and provide a basis for the development of novel cancer therapeutics.

The use of plants as medicine has always been considered. In the present study, the effect of aqueous extract collected in spring and autumn of Artemisia aucheri on the survival of HSV- 1 virus-infected cells was investigated.

In this study, the effect of aqueous extract of Artemisia aucheri in spring and autumn on the survival of Vero cell line after HSV- 1 infection at different times and concentrations was compared using TCID50 and MTT methods. The effect of aqueous extract of Artemisia aucheri at concentrations of 25, 50, 75, and 100 mg / ml on Vero cell line infected with HSV- 1 virus was performed for 24 hours in DMEM medium containing 2% (FBS) and then a dose of 50% Tissue Culture Infectious Dose(TCID50) was determined. Cell viability was also assessed by MTT assay at 2-, 0, 4, 8, and 24 hours.

The dose of 50% TCID50 with different dilutions of spring and autumn aqueous extracts of Artemisia aucheri was determined on Vero cells infected with (MOI = 0.1) HSV- 1 virus by inverted microscope and compared by MTT method. Concentrations of 50, 75, µg/ml and 100 µg/ml of spring extract and concentrations of 50 and 75 µ g / ml of autumn extract significantly reduced viral infection. Among them, spring extract with a concentration of 75 µ g / ml showed the highest viral inhibition and cell survival. Also, a significant difference was observed between spring and autumn aqueous extracts of Artemisia aucheri on cell survival (r = 0.97, P =0.0002).

The results indicated that the spring aqueous extract of Artemisia aucheri has a significant anti-HSV- 1 antiviral effect compared to the autumn aqueous extract of Artemisia aucheri (A = 0.0001) (P = 0.0001).

Colchicum speciosum is a medicinal plant rich in colchicine and phenolic compounds. In this study, the effect of different concentrations of nitrogen and nano-nitrogen fertilizers were investigated on phenols and colchicine accumulation in the corm of C.speciosum from three regions in Mazandaran province (Philband, Kelerd and Sangdarka). Five fertilizer treatments were performed. The results showed that all nitrogen and nano-nitrogen treatments caused a significant increase in corm phenol in the plants of the Philband region. Comparison of the mean flavonoids of plants in three regions in different treatments showed that the use of nitrogen and nano-nitrogen fertilizers has significantly reduced flavonoids in plant corms of all studied plants. The results showed that nitrogen fertilizer (1.1 mg / l) caused a significant increase in corm anthocyanin in all three populations. Nitrogen fertilizer with higher concentration also increased anthocyanin to some extent, while nano-nitrogen at different concentrations did not have a significant effect on the amount of anthocyanin in corm. Comparison of corm colchicine in nitrogen and nano-nitrogen treatments with control showed that nitrogen fertilizers in both concentrations decreased and nano-nitrogen fertilizers increased the amount of corm colchicine. The present results showed that the response of plants collected from the three regions to the different fertilizers was not the same.

The aim of this study was to investigate the relationship between BPA concentration and alterations of Notch1, Notch2, Notch3, Caspase-3 and Caspase-7 genes expression in the cumulus cells of infertile women with poor ovarian stimulation response (POR) following the antagonist protocol.

In this case-control study performed on 80 POR patients under 35 years of age, with 4-9 oocyte in puncture date, patients were divided into two groups. The first group consisted of 40 POR women who used plastic containers and the second group consisted of 40 POR women who used less plastic containers. Changes in the expression of Notch and Caspase genes in cumulus cells were evaluated using Q-PCR technique.

In the POR patients, the expression of Notch1-3 genes in the first group was significantly lower than the second group (p<0.05). In contrast, the expression of Caspase-3 and Caspase-7 genes in the first group was significantly higher than the second group (p<0.05).On the other hand, in both groups, was a statistically significant correlation (p<0.05) between the expression levels of Notch1-3, Caspase-3 and Caspase-7 genes and quality of Oocytes. Also, concentration of BPA in the follicular fluids of the first group was higher compared to the second group (p<0.05).

Alterations in the expression of Notch and Caspase genes in POR patients are associated with increased BPA concentration. Also, Increasing of the BPA concentration can be considered as an effective factor on the reducing of reproduction and oocyte growth.

Cytotoxic T-lymphocyte-associated protein-4 (CTLA-4) is the most important human immune checkpoint that modulates T cells activity and brings about immune-homeostasis. Accordingly, checkpoint inhibitor cancer therapy has been approved as a growing method to block over-expressed immune checkpoints, such as CTLA-4 receptors. Considering the competitive characteristics of single-domain antibodies with monoclonal antibodies, we tried to develop a camelid Nanobody against human CTLA-4. We have constructed the VHH gene library by using immunized-camel peripheral blood mononuclear cells and carrying out the Nested-PCR technique. VHH-library was screened by phage display technique and specific nanobodies against CTLA-4 protein were selected and amplified with bio-panning steps. Stronger binders were screened by Periplasmic Extract-ELISA, followed by estimating the complexity of the library. Specific anti-CTLA-4 Nanobody and 3hCTL55, with longer CDR3 and a higher binding rate, were selected for more assays. Results revealed the existence of two different clones in the library with 108 binders. In comparison with seven different antigens, using the ELISA technique confirmed the specificity of Nanobody 3hCTL55 against human CTLA-4 antigen. We calculated Nanobody 3hCTL55 affinity for human CTLA-4 antigen at 50×10-9 M, approximately. Performing western blot and Flow-cytometry techniques showed that Nanobody 3hCTL55 was able to specifically detect and attach both commercial human CTLA-4 protein and human CTLA-4 antigen on the cell surface and in the cell lysate. Taken together, this developed camelid-specific anti-CTLA-4 Nanobody 3hCTL55, selected from a high-quality immune library by phage display technique, may be effective for further study about cancer diagnosis and cancer-therapy purposes.

Raminolipids belong to the group of glycolipid bio surfactants and were first isolated from Pseudomonas aeruginosa. Rhamnolipids are a good alternative to synthetic surfactants due to their low toxicity, biodegradability and selective performance. In this study, the effect of gold (Au) nanoparticles on the growth and production of Pseudomonas aeruginosa PBCC5 bio surfactant was evaluated. Different concentrations of 1, 500 and 1000 mg / l nanoparticles were used. In this study, bio surfactant surface tension and emulsification indices (24E) were measured. The presence of nanoparticles on the bacterial surface was investigated by TEM and the morphology of nanoparticles was investigated by SEM. Binding of nanoparticles to bio surfactants was confirmed by TEM. The results showed that gold nanoparticles were not bactericidal and also increased bacterial growth and bio surfactant production. The surface tension of all samples was reduced from 72 mN / m distilled water to 35-32 mN / m.

Human papillomaviruses (HPVs) especially types 16 and 18 are known as the major causes of cervical cancer. Among HPV proteins, L1 and L2 capsid proteins, and also E7 oncoprotein are proposed as target antigens for vaccine design. In the recent years, the recombinant multiepitope polypeptides have attracted a special interest. The goal of this study is the design of L1-L2-E7 fusion construct and evaluation of its expression in bacterial expression system. 

In this study, the immunogenic and conserved epitopes of HPV16/18 L1, L2 and E7 proteins were selected using different bioinformatics analyses. After synthesis of the designed L1-L2-E7 fusion sequence in pUC57 cloning vector, its subcloning was performed in pET24a (+) prokaryotic expression vector using EcoRI/ HindIII restriction enzymes. The expression of the recombinant multiepitope polypeptide was done in E.coli Rosetta strain using IPTG inducer, and confirmed by SDS-PAGE and western blotting using anti-His-tag antibody. The expression was optimized under different conditions such as optical density (OD in wavelength of 600 nm), temperature and time after induction, and IPTG concentration.   

The recombinant pET-L1-L2-E7 vector was confirmed by the presence of a clear band (~525 bp) related to the L1-L2-E7 gene on agarose gel after enzyme digestion. The expression of L1-L2-E7 polypeptide in bacteria showed the presence of a clear band (~20 kDa) in SDS-PAGE and western blotting. The best conditions for expression of the recombinant polypeptide were at temperature of 37◦C, optical density of 0.7-0.8, IPTG concentration of 1mM, and time of 16 h after induction.  

The successful expression of the L1-L2-E7 multiepitope polypeptide was performed in bacterial system. In the next step, the recombinant polypeptide will be purified to use as a vaccine candidate.

SLC39A6 Protein (solute carrier family 39) or LIV-1 is a zinc transporter protein that is overexpressed in positive estrogen cancers such as breast cancer. The LIV-1 protein transfer zinc into the cytoplasm through the plasma membrane. Today it is known that just as a decrease in the concentration of zinc in the cell can cause cancer, an excessive increase in the concentration of zinc can also stimulate irregular cell division and caused cancer. Thus, inhibition of zinc transporter protein may play a role in preventing malignancies and metastasis. It can also be used as a diagnostic marker in the diagnosis of cancers in various laboratory methods. The present study was performed to prepare a polyclonal camel antibody for the detection of LIV-1 protein at the cell surface.

This study was started in the Pasture Institute of Iran in 2018 September and finished in February 2020. An expression construct containing the human LIV-1 gene was prepared and transferred to the E.coli BL21 by chemical (CaCl2) and heat shock method. The expression of the protein was induced by IPTG and then protein was purified by affinity (Ni-NTA) chromatography. After preparing recombinant protein one female camel was immunized, 6 times at two weeks intervals with Freundchr('39')s adjuvant. After immunization, the isolated polyclonal antibody was evaluated by ELISA, western blotting and flow cytometry in the detection of LIV-1 protein.

The result showed that LIV-1 protein was well purified and also the camel polyclonal antibody was able to detect LIV-1 protein in ELISA, western blot and also it can detect LIV-1 on the cell surface as shown by flow cytometry test.

In recent years, LIV-1 has been shown to be a good candidate as a marker in breast cancer, so polyclonal antibodies against LIV-1 can be used for early detection of breast cancer by various diagnostic methods. In this study, it has been shown that polyclonal camel antibodies can be used in laboratory methods and can be considered for immunological tests and therapeutic applications.