فصلنامه تازه های بیوتکنولوژی سلولی مولکولی
پیاپی 44 (پاییز 1400)

Herpes simplex virus type 2 (HSV-2) is one of the most important human pathogenic viruses. The virus causes genital herpes and can cause miscarriage, pregnancy complications, and infertility in women. The aim of this study was to evaluate the role of HSV-2 in idiopathic abortion by PCR.

Fifty blood serum samples of women with idiopathic miscarriages were collected and transferred to the laboratory. The PCR test was performed using standard HSV-2 strain DNA and then the limit of detection (LOD) and specificity of the test was determined using a different strain of DNA. A standard test was performed using positive and negative controls on the samples and the results were analyzed by agarose gel electrophoresis.

The PCR technique est optimized and 231 bp amplicon were observed in agarose gel. In specificity test only with DNA HSV-2 achieved positive results as well as a limit of detection 10 Copy / Reaction. Of the 50 samples studied, 9 (4.5%) were positive.

Idiopathic miscarriages are miscarriages that do not seem to have a definite cause. But clearly, several factors, including infectious agents that are difficult to identify, can cause miscarriage. One of these factors could be HSV-2. The results of this study showed that HSV-2 is one of the most important factors in this type of miscarriage and that HSV-2 can be quickly identified by tests such as PCR.

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).

Microorganisms are more important than other sources in dye production because microorganisms have advantages such as rapid growth, higher yields, and easier extraction. Natural pigments are healthier than chemical types and their use is useful in various industries. The aim of this study was to isolate and extract the pyocyanin pigment from Pseudomonas aeruginosa isolates in order to preparation of colored paper and candle.

In this study, sampling was performed from different hospital environments in order to isolate P. aeruginosa. Environmental isolates of P. aeruginosa were identified using morphological features and differential biochemical methods. Confirmatory identification was performed by tracing the pbo1 gene using colony PCR. After extracting pyocyanin pigment with chloroform, finally, the produced pyocyanin was used to prepare colored paper and candle and the optical stability of pyocyanin was investigated.

From 43 environmental samples collected, 15 isolates of P. aeruginosa were identified and confirmed by molecular method. The mean of optical absorption (OD520) of pyocyanin after extraction was 0.488. Staining of paper and candle with the extracted pyocyanin was successful and the optical stability of pyocyanin was favorable.

Due to the rapid growth of bacteria and the possibility of its mass production in a short time and based on the results of this study, bacterial pigments can be used in various industries, including paper industry, and replaced by chemical dyes.

Wild plants in nature are valuable samples for use in various studies due to their characteristics such as resistance to adverse environmental conditions, resistance to pests and diseases and even having high active ingredients. Today, with the increasing resistance of bacteria to antibiotics, the use of medicinal plants has been welcomed.The  present study aimed  to  investigate  the  effect  of antibacterial effects of different ecotypes of Verbascum thapsus extract.

Verbascum thapsus from three regions of North Khorasan (Ashkhaneh, Bojnourd and Shirvan), after collection and drying, was extracted by methanol solvent by soaking and its  antibacterial properties was investigation against( Escherichia coli, Bacillus cereus and Staphylococcus aureus) using disk and well diffusion , MIC (Minimum Inhibitory Concentration ) and MBC ( Minimum Bactericidal  Concenteration) methods.

The results showed that between different ecotypes of North Khorasan, Verbascom extract of Bojnourd region had the most inhition zone  in disk and well diffusion method on Staphylococcus aureus bacteria were equal to 12.33 and 14.66 mm, respectively, and for Bacillus cereus bacteria were 11.33 and 13.66 mm, respectively And MIC and MBC of Verbascum extract of Bojnourd region on bacteria (Staphylococcus aureus) 12. mg/ml were observed as the most sensitive bacteria.

In this study, it was found that among different ecotypes of North Khorasan, methanolic extract of Verbascum thapsus of Bojnourd region had the most antibacterial effect on bacteria (Bacillus cereus and Staphylococcus aureus) however the extract was not effective on Escherichia coli.

Silybinin, one of the active pharmaceutical ingredients in some medical herbs, has been proven to have anti-cancer effects. The aim of this study was to synthesis and characterize lipid nano-carriers containing silybinin in order to breast cancer treatment.

Lipid nanocarriers were synthesized using a thin film method and the drug was loaded on them. Then the physicochemical properties were evaluated using DLS, AFM and spectrophotometers and their toxicity was measured in comparison with the free form of the drug by MTT assay on BT-474 breast cancer cell line.

The synthesized nano-system with a size of 113.2 nm, zeta potential of -22 ± 1.63 mV, slow release type with appropriate physicochemical properties improves the anti-cancer effects of drug on breast cancer cells.

The results of this study demonstrated that the synthesized system, in addition to having appropriate physicochemical properties, could improve anti-cancer effect of drug.

Immunodeficiency virus-1 (HIV-1) is a global health problem and it has affected more than 75 million individuals since the epidemic started. Various approaches have been investigated to some up with a preventive or therapeutic formulation against this virus. However, none of them has been achieved. The aim of the study was the immunological evaluation of HIV-1 Nef-MPER-V3 Harboring IMT-P8 penetrating peptide in BALB/c mice to induce effective immune responses.

In current study, to evaluate of the antigen immunogenicity of IMT-P8-Nef-MPER-V3, 11 different groups of female BALB/c mice were immunized with three doses of the antigen with or without Hsp27 and Hp91 adjuvants formulation. The immune responses were assessed using IgG ELISA assay and its isotype determination. IL-10 and IFN-γ were also evaluated by sandwich ELISA.

The data showed that the recombinant protein developed different levels of humoral and cellular responses. IMTP8-Nef-MPER-V3+Hp91 groups reached highest IgG2a, and elicited strong IFN-γ production towards a Th1 response compared to the other groups. Cytokine assay indicated that the immunized mice with the antigen formulation containing IMT-P8 CPP applied with Hp91 has a high potency in immune induction.

These results demonstrated that application of IMTP8-Nef-MPER-V3 combining the adjuvant-formulated (Hp91) provides strong responses which must be considered as an effective formulation towards a potential HIV vaccine candidate.

Staphylococcus aureus is one of the most important pathogens in humans. The development of resistance to various antibiotics in this bacterium has become a global problem that due to the importance accessory gene regulator (agr) in this bacterium, which is responsible for coordinating and controlling the production and secretion of toxins and virulence factors. The purpose of this study was to investigate the pattern of antibiotic resistance and determine specific groups agr in order to find the dominant type of region in sensitive and methicillin-resistant Staphylococcus aureus.

Different clinical samples were prepared of Clients to Imam Reza Hospital in Bojnourd and phenotypic tests were performed on Staphylococcus aureus strains. Disc diffusion method was used to confirm drug sensitivity. DNA extraction was performed using a QiaAmp kit and Methicillin-resistant strains with mecA gene were identified by polymerase chain reaction (PCR) and agr groups were determined by multiplex PCR.

In this study, the highest antibiotic resistance of Staphylococcus aureus against cefoxitin was (75%). Isolates (73. %) have mecA gene and in sensitive and methicillin resistant strains the highest percentage belongs to agrI group with frequency of 60.71% and 15.58%.

Due to the fact that the pathogenicity of strains belonging to agr groups is different in various regions, it is necessary to determine the strains isolated from each region in order to find the dominant type in that region.

Human Papillomaviruses (HPV) infection was known to be the leading cause of cervical cancer. Cervical cancer is the fourth most common cancer among women in the world. Designing a DNA vaccine with therapeutic properties as well as prevention can have a significant impact on reducing morbidity and mortality.

Gene sequences including the immunogenic and conserved epitopes of L1 and E7 proteins of high risk papillomaviruses were designed. After synthesis of the L1-E7 fusion construct in pUC57 cloning vector, its subcloning was performed in pcDNA3.1 (-) eukaryotic vector. The concentration and purity of the recombinant pcDNA-L1E7 plasmid was determined by NanoDrop spectrophotometry.  

The gene sequence after cutting from pUC57-L1E7 by BamHI/HindIII restriction enzymes, was subcloned in the same cloning site of pcDNA3.1 (-) vector. The presence of gene was confirmed by digestion with BamHI/HindIII enzymes, as a 639 bp fragment on 1% agarose gel. The recombinant pcDNA-L1E7 plasmid was purified by endotoxin-free extraction kit for isolation of bacterial lipopolysaccharide. The concentration of the purified DNA was obtained about 1474 ng/ µl.

Cloning of the L1-E7 fusion construct in the eukaryotic vector was successful. In the next steps, the recombinant vector will be used for gene vaccine studies in vivo.