The role of GlcNAc-PI-de-N-acetylase gene by gene knockout through homologous recombination and its consequences on survival, growth and infectivity of Leishmania major in in vitro and in vivo conditions

At present, there are no efficacious vaccines or effective drugs against leishmaniasis; therefore new and innovative control methods are urgently required. One way to achieve this important goal is through using reverse genetic engineering to evaluate important enzymes, proteins and macromolecules. One of the most important enzymes for Glycosylphosphatidylinositol (GPI) biosynthetic pathways is GlcNAc-PI-deN-acetylase (GPI12). The molecular constructs were cloned in Escherichia coli strain Top 10 and confirmed by molecular methods and were transfected by electroporation into Leishmania major. We demonstrated that two alleles of the GPI12 gene in L. major were successfully removed and enabling the generation of a null mutant, which supports the idea that GPI12 is not an essential gene for the growth and survival of Leishmania and the homozygous knockouts of Leishmania are able to survive. We were able to produce a mutant parasite that caused no damaged to the host. Further investigations are essential to check the safety profile in laboratory animal

Indoor environment assessment of special wards of educational hospitals for the detection of fungal contamination sources: A multi-center study (2019-2021)

Background and Purpose: The hospital environment was reported as a real habitat for different microorganisms, especially mold fungi. On the other hand, these opportunistic fungi were considered hospital-acquired mold infections in patients with weak immune status. Therefore, this multi-center study aimed to evaluate 23 hospitals in 18 provinces of Iran for fungal contamination sources.Materials and Methods: In total, 43 opened Petri plates and 213 surface samples were collected throughout different wards of 23 hospitals. All collected samples were inoculated into Sabouraud Dextrose Agar containing Chloramphenicol (SC), and the plates were then incubated at 27-30ºC for 7-14 days.Results: A total of 210 fungal colonies from equipment (162, 77.1%) and air (48,22.9%) were identified. The most predominant isolated genus was Aspergillus (47.5%),followed by Rhizopus (14.2%), Mucor (11.7%), and Cladosporium (9.2%). Aspergillus(39.5%), Cladosporium (16.6%), as well as Penicillium and Sterile hyphae (10.4% each), were the most isolates from the air samples. Moreover, intensive care units (38.5%) and operating rooms (21.9%) had the highest number of isolated fungal colonies. Out of 256 collected samples from equipment and air, 163 (63.7%) were positive for fungal growth.The rate of fungal contamination in instrument and air samples was 128/213 (60.1%) and 35/43 (81.2%), respectively. Among the isolated species of Aspergillus, A. flavus complex (38/96, 39.6%), A. niger complex (31/96, 32.3%), and A. fumigatus complex (15/96, 15.6%) were the commonest species.Conclusion: According to our findings, in addition to air, equipment and instrument should be considered among the significant sources of fungal contamination in the indoor environment of hospitals. Airborne fungi, Hospital, Indoor air, Equipment, Sources of fungal contamination in the indoor environment of hospitals

Comparing postpartum stressors and social support level in primiparous and multiparous women

Background and aim: Postpartum period is an exclusive period after birth which can act as a potential stressor and could be accompanied with psychological disorders. Social support could play an important role in maternal mental health. Considering various stressors and different levels of social support for women, this study aimed to compare postpartum stressors as well as social support level between primiparous and multiparous women. Method: This descriptive comparative study was conducted on 400 primiparous and multiparous mothers who referred to urban health centers, Mashhad, Iran in 2011. They had no history of medical or psychological problems and had healthy term neonates who were 8-25 days old. The sampling was carried out through a multistage cluster sampling. Data were collected using modified Hung questionnaire of postpartum stressors, Helen questionnaire of anxiety and Hopkins social support questionnaire. Data were analyzed using descriptive analytic statistics by SPSS version 11. Results: The mean stress scores were 242.5±157.1 in primiparous and 28.8±179.8 in multiparous women. The main stressor of primiparous and multiparous women was neonate bathing and lower-back pain, respectively. The mean score of social support was 108.3±8.25 in the primiparous and 102.0±26.6 in multiparous women, which showed a significant difference between two groups (P=0.000). Conclusion: Various care programs are essential to support primiparous and multiparous mothers from different stressors that they face in postpartum period. It is also recommended to provid more information regarding the social support for the families

Designing and Cloning Molecular Constructs to Knock Out N-Acetylglucosamine Phosphatidylinositol De-N-Acetylase (GPI12) Gene in Leishmania major (MRHO/IR/75/ER)

Background: Leishmaniasis represents a major public health concern in tropical and sub-tropical countries. At present, there is no efficacious vaccine against the disease and new control methods are needed. One way to access this important goal is to knock out genes of specific macromolecules to evaluate the effect of deletion on the growth, multiplication, pathogenesis and immunity of the parasite. The aim of this study was to design and clone molecular constructs to knock out N-acetylglucosamine phosphatidylinositol de-N-acetylase (GPI12) gene in Leishmania major. Methods: For designing and making molecular constructs, we used pLEXSY-neo2 and pLEXSY-hyg2 vectors. The molecular constructs were cloned in E. coli strain Top10. The molecular constructs were transfected by electroporation into L. major in two stages. Results: The molecular constructs were confirmed by Colony PCR and sequencing. The recombinant strains were isolated by selective antibiotics, after which they were confirmed by PCR, Southern and Western blots. Conclusion: Recombinant parasites were created and examined for subsequent study. With the use of molecular constructs, it was possible to remove and study gene GPI12 and to achieve a live recombinant Leishmania parasite that maintained the original form of the antigenic parasites. This achievement can be used as an experimental model for vaccine development studies. Further investigations are essential to check this model in a suitable host