Evaluation of Chitosan Nanoparticle Antimicrobial Effect on Isolated Listeria monocytogenes Bacteria from Pregnant Women and L. monocytogenes ATCC 7644.

Background: Listeria monocytogenes is a gram positive, facultative intracellular bacteria and it is a causative agent of listeriosis. Abortion is one the most important side effect of listeriosis. Nano chitosan is widely used as nanomaterials considered due to its characteristics such as bactericidal and nontoxicity activity. The aim of this study was isolation of L. monocytogenes bacteria from pregnant women vaginal samples and evaluation of chitosan nanoparticles effects against them. Methods: Overall, 100 vaginal specimens were collected from pregnant women with and without a history of abortion referred to Tehran's Hospitals from Sep 2019 to Jul 2020 with questionnaires. Then, using biochemical methods, L. monocytogenes bacteria were isolated and identified. Isolated L. monocytogenes strains were confirmed using PCR and evaluation of prfA gene, which is the main gene for identification of this bacterium. The effect of chitosan nanoparticles was evaluated in comparison with the antibiotics used based CSLI guideline on isolated bacteria by well diffusion method. MIC and MBC were determined for nanoparticle at the end. Results: Five strains of L. monocytogenes that were confirmed by PCR method. Moreover, a statistically significant relationship was observed between the isolated strains and the samples taken from women with a history of abortion. MIC and MBC for L. monocytogenes ATCC 7644 were 156.25 and 312.5 µg/ml and for 5 isolated strains were 78.12 and 158.25 µg/ml, respectively. Conclusion: L. monocytogenes could be a causative agent of abortion in pregnant women. Concerning the results, Nano chitosan has acceptable antibacterial activity against L. monocytogenes.

Isolation and identification of carotenoid-producing Rhodotorula sp. from Pinaceae forest ecosystems and optimization of in vitro carotenoid production.

Yeasts are alternative source of natural carotenoids, a group of colored terpenoids with various market applications. Carotenoid Production by yeast fermentation technology is greatly effective and proposes considerable benefits with large scale production, cost effectiveness and safety. In this study, four pigment-producing yeasts were isolated from forest park soils with the potential to produce carotenoids. Morphological, physiological, biochemical and molecular characterization indicates the isolates belong to Rhodotorula mucilaginosa. Carotenoid production was optimized by small scale cultivation. The optimum condition was 120 h of incubation at pH 6.0, 28 °C, white light irradiation, Yeast Extract Peptone Glycerol medium composed of 10 g/L yeast extract, 20 g/L peptone, 20 ml/L glycerol, yielding maximum content of 223.5 µg/g dry weight. The ß-carotene content was confirmed by HPLC and FT-IR. The results suggested that soil yeasts are potential sources of carotenoids that could be utilized as a natural agent for industrial products.

Human Cutaneous Leishmaniosis in Iran, Up to Date-2019.

Cutaneous leishmaniasis is a neglected and parasitic vector borne diseases that is endemic in tropical and subtropical countries, including Iran. The aim of this study was to explain the present status of CL in Iran. This report is based on data that recorded by cutaneous leishmaniasis surveillance system in 2019, and evaluated in Center for Communicable Diseases Management in Ministry of Health in Iran. Iran has been considered an endemic area for cutaneous leishmaniasis in the world. Dependent to activities for cutaneous leishmaniasis control the number of cases decreased from 23202 in 2008 (Incidence rate 32 per 100000) to 13124 in 2019 (Incidence rate 15.8 per 100000), more cases reported from September to December, in 2019, 46% of cases had one lesion and 21% had 2 lesions, 85% of cases diagnosed when the diameter of lesions had 3 centimeters and bellows. Although the Leishmania control program began in 1977, the incidence of the disease has dropped dramatically since 2008 when the new cutaneous leishmaniasis control program have been implemented. Although in some areas the incidence of the disease increased, but the implementation of the new program has reduced the number of cases, in order to continue reducing the disease, permanent support for the control programs is needed.

Designing novel construction for cell surface display of protein E on Escherichia coli using non-classical pathway based on Lpp-OmpA.

Today, transference of recombinant protein on the outer surface of bacteria is deemed as a valuable process for various applications in biotechnology including preparation of vaccines. In this study, Lpp-OmpA structure was used to present outer membrane protein E of Haemophilus influenzae on E. coli outer membrane. Also, a structure was designed according to Lpp-OmpA based on non-classical secretion pathway using bioinformatics software such as MEMSAT-SVM, ScrotumP and SignalP where it lacked any signal peptide at its N-terminal. Potential of this structure in the presentation of protein E on the surface of E. coli through non-classical pathway was indicated by western blotting, SDS page and fluorescent microscopy techniques, similarly its effectiveness was compared with Lpp-OmpA system. The results of the current study showed that the new structure had higher efficiency than Lpp-OmpA, and it could transport protein E on outer membrane well. This study is the first report in the presentation of H. influenzae PE onto the surface of E. coli by Lpp-OmpA, and the structure originated from Lpp-OmpA, according to the non-classical secretion pathway. Our results suggest that non-classical secretion pathway may be exploited as a new secretory pathway on the outer surface of the cell for recombinant proteins.

Estrogen treatment enhances neurogenic differentiation of human adipose derived stem cells in vitro.

OBJECTIVES: Estrogen is a sexual hormone that has prominent effects on reproductive and non-reproductive tissues. The aim of this study is to evaluate the effects of estrogen on the proliferation and neural differentiation of human adipose derived stem cells (ADSCs) during neurogenic differentiation. MATERIALS AND METHODS: Isolated human ADSCs were trans-differentiated in neural induction medium containing neurobasal medium, N2 and B27 with or without 17ß-estradiol (E2) treatment. Proliferation rate and neural differentiation of human ADSCs were assessed using MTT assay, immunostaining and real time RT- PCR analysis, respectively. RESULTS: Analysis of data show that estradiol treatment can significantly increase proliferation rate of differentiated cells (P<0.05). Immunocytochemical and real time RT-PCR analysis revealed that the expression of precursor and mature neuronal markers (nestin and MAP2) was significantly higher in the E2 treated cell cultures when compared to the untreated cell cultures (P<0.05). CONCLUSION: According to our findings, estrogen can promote proliferation and neuronal differentiation of human ADSCs.

Co-culture with neurotrophic factor secreting cells induced from adipose-derived stem cells: promotes neurogenic differentiation.

Adipose-derived stem cells (ADSCs) and bone marrow stem cells (BMSCs) can be equally proper in the treatment of neurodegenerative diseases. However, ADSCs have practical benefits. In this study, we attempted to induce the secretion of neurotrophic factors (NTF) in human ADSCs. We then evaluated the effects of co-culture with NTF secreting cells in neural differentiation of human ADSCs. Isolated human ADSCs were induced to neurotrophic factors secreting cells. To evaluate the in vitro effects of NTF-secreting ADSCs on neurogenic differentiation of ADSCs, we used neurogenic induction medium (control group), the combination of neurogenic medium and conditioned medium, or co-cultured NTF-secreting ADSCs which were encapsulated in alginate beads (co-culture) for 7 days. ELISA showed increased (by about 5 times) release of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in NTF-secreting ADSCs compared to human ADSCs. Real time RT-PCR analysis revealed that NTF-secreting ADSCs highly expressed NGF and BDNF. In addition, co-culture with NTF-secreting ADSCs could also promote neuronal differentiation relative to gliogenesis. Overall, NTF-secreting ADSCs secrete a range of growth factors whose levels in culture could promote neuronal differentiation and could support the survival and regeneration in a variety of neurodegenerative diseases.

Comparing brain-derived neurotrophic factor and ciliary neurotrophic factor secretion of induced neurotrophic factor secreting cells from human adipose and bone marrow-derived stem cells.

Adipose derived stem cells (ADSCs) and bone marrow stem cells (BMSCs) may be equally beneficial in treating neurodegenerative diseases. However, ADSCs have practical advantages. In this study, we aimed to induce neurotrophic factors secreting cells in human ADSCs. Then, we compared the level of brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) secretion in neurotrophic factors secreting cells from human adipose and bone marrow-derived stem cells. Isolated human ADSCs and BMSCs were induced to neurotrophic factor (NTF)-secreting cells. The levels of expression and secretion of BDNF and CTNF of induced cells were assessed using immunocytochemical, Real-Time polymerase chain reaction, and enzyme linked immunosorbent assay (ELISA). The level of BDNF significantly increased in both the induced mesenchymal stem cells (MSCs) relative to ADSCs and the BMSCs (P < 0.01). Moreover, ELISA analysis showed that the release of BDNF in the induced BMSCs was almost twofold more than the induced ADSCs. Overall, NTF-secreting factor cells derived BMSCs and ADSCs could secret a range of different growth factors. Therefore, the variation in neurotrophic factors of different induced MSC populations suggest the possible beneficial effect of each specific kind of neurotrophic factor secreting cells for the treatment of a particular neurodegenerative disease.

Portable, battery-operated, low-cost, bright field and fluorescence microscope.

This study describes the design and evaluation of a portable bright-field and fluorescence microscope that can be manufactured for $240 USD. The microscope uses a battery-operated LED-based flashlight as the light source and achieves a resolution of 0.8 microm at 1000x magnification in fluorescence mode. We tested the diagnostic capability of this new instrument to identify infections caused by the human pathogen, Mycobacterium tuberculosis. Sixty-four direct, decontaminated, and serially diluted smears were prepared from sputa obtained from 19 patients suspected to have M. tuberculosis infection. Slides were stained with auramine orange and evaluated as being positive or negative for M. tuberculosis with both the new portable fluorescence microscope and a laboratory grade fluorescence microscope. Concordant results were obtained in 98.4% of cases. This highly portable, low cost, fluorescence microscope may be a useful diagnostic tool to expand the availability of M. tuberculosis testing at the point-of-care in low resource settings.

Coagulase-negative staphylococci isolated from ocular wound infections after laser refractive surgery: attachment to and accumulation on soft contact lenses.

OBJECTIVES: Disposable soft contact lenses that are commonly used after laser refractive surgery are known to be colonized by bacteria and play a key role in Bacterial Keraitis (BK) pathogenesis. Coagulase-negative staphylococci (CoNS) have been found to be the most common pathogen involved in this postoperative infection. In this study a rapid and a simple assay was developed for studying attachment and accumulation of CoNS on soft contact lenses in vitro using [3H] thymidine. METHODS: Thirty-five isolates of CoNS were obtained from 27 laser refractive surgery patients. Twenty-five of these thirty-five CoNS were isolated in multiple cultures. Ten CoNS were isolated in cultures from patients who underwent reoperation. The assay was optimized using a biofilm-producing strain, S. epidermidis RP62A, which was subcultured overnight at 37 degrees C on blood agar medium. Quantitative determination of biofilm production was tested. Presence of the genes icaADB and icaD was determined in all isolates. All isolates were biochemically analyzed using the Phene Plate (PhP) system modified for typing of CoNS. The CoNS isolates were further characterized to species level using ID32Staph.Mann-Whitney rank sum test and chi-square test were used to identify statistical differences in adherence, index, antibiotic susceptibility patterns, and biofilm production or presence of the ica operon between clinically significant isolates and non-postoperative BK isolates. RESULTS: No differences in attachment and accumulation were found between isolates causing BK after laser refractive surgery and contaminant isolates. In addition, there were no differences in the distribution of the ica operon between the two groups, as determined by polymerase chain reaction. Nevertheless, the ability to produce biofilm was found to be present significantly more frequently among BK isolates than among non-postoperative BK isolates. CONCLUSIONS: This study shows that the method using radioactive thymidine to analyze adherence of CoNS to soft contact lenses enables detection of differences in the adherence patterns of individual isolates.

Investigation of Yersinia pestis in Xenopsylla astia.

In this study, at the Department of Parasitology in the Pasteur Institute of Iran, Xenopsylla were allowed to feed on mice infected with Yersinia pestis. After 24-48 hours, they were killed by ether and kept in alcohol (70%) for 20 minutes. They were then examined for pathological signs and bacilli in different tissues and organs, longitudinally and cross-sectionally. The samples were studied using conjugated antibody and fluorescence microscopy. The results of this study revealed that the bacilli are abundant in the proventriculus after 6 hours, but it was found in other organs, rarely.