Somatic embryogenesis and plantlet regeneration from protoplast culture of Muscari neglectum Guss

A protocol has been developed for plant regeneration from protoplast culture of Muscari neglectum using regenerable embryogenic calli obtained from bulb culture on Murashige and Skoog medium (MS) plant growth regulators (PGR)-free or containing lower 1-naphthaleneacetic acid (NAA) and 6- benzylaminopurine (BA) concentrations and 30 g/l sucrose. Protoplasts were isolated directly from embryogenic calli, embedded in Ca-alginate beads and cultured with nurse cells in MS medium supplemented with 1 mg/l each of NAA and BA, 100 mg/l ascorbic acid and 0.5 M mannitol at 25°C in darkness. After 4 weeks of culture, microcalli appeared on the surface of the Ca-alginate beads. Growth of microcalli in the medium with nurse cells (33.3%) was much better than those in the medium without nurse cells (6.5%). Transferring beads onto MS medium supplemented with 0.1 mg/l BA increased the growth of embryogenic calli. Somatic embryo development was observed either on half strength MS medium PGR-free or with 1 mg/l abscisic acid at 25°C under continuous illumination with fluorescent light. Maturated embryos germinated and then converted to plantlets on half strength MS medium containing 1 mg/l BA after 3 months. The plantlets left in the medium produced bulbs after 5 months.Key words: Ca-alginate beads, Muscari neglectum, nurse culture, plantlet regeneration, protoplast culture, somatic embryogenesis

First record of two hard coral species (Faviidae and Siderastreidae) from Qeshm Island (Persian Gulf, Iran)

Abstrak. Moradi M, Kamrani E, Shokri MR, Ranjbar MS, Hesni MA (2009) Rekaman pertama dua spesies karang keras (Faviidae dan Siderastreidae) dari Pulau Qeshm (Teluk Persia, Iran). Nusantara Bioscience 2: 34-37. Dua jenis karang keras termasuk Cyphastrea chalcidicum (Forskal 1775) (Faviidae) dan Coscinaraea monile (Forskal 1775) (Siderastreidae) dikumpulkan dari selatan Pulau Qeshm (Teluk Persia, Iran) pada akhir tahun 2008. Spesies ini sebelumnya dilaporkan terdapat di Teluk Persia selatan, Teluk Aden, Afrika Tenggara dan Indo-Pasifik. Tinjauan literatur pada distribusi kedua jenis mengungkapkan bahwa spesies ini pertama kali tercatat dari Teluk Persia. Temuan ini semakin menunjukkan tingginya keragaman fauna karang di perairan Iran di bagian utara Teluk Persia. Kata kunci: catatan pertama, Coscinaraea monile, Cyphastrea chalcidicum, Qeshm island, Persian gulf

Novel 5-oxo-hexahydroquinoline derivatives: design, synthesis, in vitro P-glycoprotein-mediated multidrug resistance reversal profile and molecular dynamics simulation study

Overexpression of the efflux pump P-glycoprotein (P-gp) is one of the important mechanisms of multidrug resistance (MDR) in many tumor cells. In this study, 26 novel 5-oxo-hexahydroquinoline derivatives containing different nitrophenyl moieties at C-4 and various carboxamide substituents at C-3 were designed, synthesized and evaluated for their ability to inhibit P-gp by measuring the amount of rhodamine 123 (Rh123) accumulation in uterine sarcoma cells that overexpress P-gp (MES-SA/Dx5) using flow cytometry. The effect of compounds with highest MDR reversal activities was further evaluated by measuring the alterations of MES-SA/Dx5 cells' sensitivity to doxorubicin (DXR) using MTT assay. The results of both biological assays indicated that compounds bearing 2-nitrophenyl at C-4 position and compounds with 4-chlorophenyl carboxamide at C-3 demonstrated the highest activities in resistant cells, while they were devoid of any effect in parental nonresistant MES-SA cells. One of the active derivatives, 5c, significantly increased intracellular Rh123 at 100 mu M, and it also significantly reduced the IC50 of DXR by 70.1% and 88.7% at 10 and 25 mu M, respectively, in MES-SA/Dx5 cells. The toxicity of synthesized compounds against HEK293 as a noncancer cell line was also investigated. All tested derivatives except for 2c compound showed no cytotoxicity. A molecular dynamics simulation study was also performed to investigate the possible binding site of 5c in complex with human P-gp, which showed that this compound formed 11 average H-bonds with Ser909, Thr911, Arg547, Arg543 and Ser474 residues of P-gp. A good agreement was found between the results of the computational and experimental studies. The findings of this study show that some 5-oxo-hexahydroquinoline derivatives could serve as promising candidates for the discovery of new agents for P-gp-mediated MDR reversal

A Time Series Model for Assessing the Trend and Forecasting the Road Traffic Accident Mortality

Background: Road traffic accident (RTA) is one of the main causes of trauma and known as a growing public health concern worldwide, especially in developing countries. Assessing the trend of fatalities in the past years and forecasting it enables us to make the appropriate planning for prevention and control. Objectives: This study aimed to assess the trend of RTAs and forecast it in the next years by using time series modeling. Materials and Methods: In this historical analytical study, the RTA mortalities in Zanjan Province, Iran, were evaluated during 2007 - 2013. The time series analyses including Box-Jenkins models were used to assess the trend of accident fatalities in previous years and forecast it for the next 4 years. Results: The mean age of the victims was 37.22 years (SD = 20.01). From a total of 2571 deaths, 77.5% (n = 1992) were males and 22.5% (n = 579) were females. The study models showed a descending trend of fatalities in the study years. The SARIMA (1, 1, 3) (0, 1, 0) 12 model was recognized as a best fit model in forecasting the trend of fatalities. Forecasting model also showed a descending trend of traffic accident mortalities in the next 4 years. Conclusions: There was a decreasing trend in the study and the future years. It seems that implementation of some interventions in the recent decade has had a positive effect on the decline of RTA fatalities. Nevertheless, there is still a need to pay more attention in order to prevent the occurrence and the mortalities related to traffic accidents

Prevalence of Chlamydia trachomatis infections in symptomatic women by polymerase chain reaction (PCR) immunofluorescence and Giemsa stain

Chlamydia trachomatis is a ubiquitous human pathogen that is responsible for the most prevalent bacterial sexually transmitted disease worldwide. Studies show that polymerase chain reaction (PCR) is more sensitive than cellular culture for detection of C. trachomatis infections. The aim of this study is to compare different laboratory methods, including Giemsa staining, direct immunofluorescence assay (DFA) and PCR for detection of C. trachomatis in women with urethral symptoms. In this study, 130 women with urethral symptoms admitted in the gynecology clinic, were used and specimens were obtained with endocervical swab for Giemsa staining, DFA and PCR. All the cases underwent these three techniques. Demographic data and the medical history of patients were obtained by direct interview; however, the mean age of cases was 33.8±9.06. Clinical symptoms included abnormal vaginal discharge in 101 cases (77.7%), spotting in 14 cases (10.8%), dysmenorrheal in 7 cases (5.4%), irritation in 6 cases (4.6%) and dysuria in 2 cases (1.5%). In DFA technique, 5 cases (3.8%) were positive and 3 (2.3%) were suspicious, while in the PCR technique, 6 cases (4.6%) were positive for C. trachomatis. However, 3 suspicious cases with DFA were negative in PCR. There was no positive case for C. trachomatis in Giemsa staining. In conclusion, C. trachomatis was not frequent in this study and it can be concluded that this infection was not a major hygienic problem in the same populations that were previously studied. Consequently, the causes that necessitate monogamy could be related to religious causes. Frequency of Chlamydia detection of DFA and PCR was same in the two groups. Nonetheless, Giemsa staining is not a reliable method for evaluating C. trachomatis.Key words: Chlamydia trachomatis, polymerase chain reaction (PCR), direct immunofluorescence assay (DFA)

Typing methods used in the molecular epidemiology of microbial pathogens: a how-to guide

Microbial typing is often employed to determine the source and routes of infections, confirm or rule out outbreaks, trace cross-transmission of healthcare-associated pathogens, recognize virulent strains and evaluate the effectiveness of control measures. Conventional microbial typing methods have occasionally been useful in describing the epidemiology of infectious diseases. However, these methods are generally considered too variable, labour intensive and time-consuming to be of practical value in epidemiological investigations. Moreover, these approaches have proved to be insufficiently discriminatory and poorly reproducible. DNA-based typing methods rely on the analysis of the genetic material of a microorganism. In recent years, several methods have been introduced and developed for investigation of the molecular epidemiology of microbial pathogens. Each of them has advantages and limitations that make them useful in some studies and restrictive in others. The choice of a molecular typing method therefore will depend on the skill level and resources of the laboratory and the aim and scale of the investigation. This study reviews the most popular DNA-based molecular typing methods used in the epidemiology of bacterial pathogens together with their advantages and limitations

Detecting very low allele fraction variants using targeted DNA sequencing and a novel molecular barcode-aware variant caller

The supplementary materials include supplementary methods, supplementary figures, and supplementary tables. (PDF 706 kb

Characterization of endemic Shigella boydii strains isolated in Iran by serotyping, antimicrobial resistance, plasmid profile, ribotyping and pulsed-field gel electrophoresis

Background: Shigellosis is one of the major causes of morbidity in children with diarrhea in Iran. The present study was undertaken to characterize apparently sporadic Shigella boydii strains isolated from pediatric patients in Iran. Findings: Ten S. boydii strains isolated from pediatric cases of gastroenteritis and acute diarrhea in Tehran between December 2002 and November 2003 were submitted to serotyping, antimicrobial susceptibility testing, plasmid profile analysis, ribotyping and pulsed field gel electrophoresis (PFGE). Seven isolates were attributed to serotype 2, whereas the remaining three belonged to serotypes 14, 18, 19, respectively. Six drug resistance phenotypes (R1 to R6) were defined with R4 - streptomycin (STR), ampicillin (AMP), sulfamethoxazole-trimethoprim (SXT) - being the most prevalent. Plasmid analysis resulted in seven different plasmid profiles with one to five DNA bands. All strains, but one, shared the same ribotype, but PFGE differentiated them in four groups. Conclusion: Based upon ribotyping and PFGE results, endemic circulation of S. boydii in Tehran, Iran, could be attributed to a few clones. Resistance pattern and plasmid profile analysis proved to be very effective in discriminating apparently unrelated strains of S. boydi

Microencapsulation of Bacillus velezensis Using Alginate-Gum Polymers Enriched with TiO2 and SiO2 Nanoparticles

Bacillus bacteria are a group of plant growth stimulants that increase plant growth and resistance to plant pathogens by producing various metabolites. With their large surface area and small size, nanoparticles can be used in controlled-release formulations and increase the efficiency of the desired product. Encapsulation of biological agents in combination with nanoparticles can be an essential step in increasing the performance of these agents in adverse environmental conditions. In this study, which is the result of a collaboration between scientists from Italy and Iran, Bacillus velezensis was encapsulated in alginate combined with whey protein and zedo, mastic, and tragacanth gums in the presence of silica and titania nanoparticles to obtain two-layer and multilayer assemblies acting as novel, smart micro-encapsulation systems. The results of laboratory studies showed that the B. velezensis could produce protease, lipase, siderophore, auxin, and a dissolution of mineral phosphate. Scanning electron microscopy images (SEM) showed that the studied microcapsules were almost spherical. Moisture affinity, swelling, and efficiency of each microcapsule were examined. The results showed that the highest encapsulation efficiency (94.3%) was related to the multilayer formulation of alginate-whey protein-zedo. XRD and FTIR spectroscopy showed that the alginate, whey protein, and zedo were mixed properly and no incompatible composition occurred in the reaction. This study aimed to provide a suitable formulation of biofertilizers based on biodegradable compounds as an alternative to chemical fertilizers, which is low cost and very effective without harming humans and the environment