IDENTIFICATION OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS USING TOUCHDOWN PCR AND PHENOTYPIC METHODS FROM PATIENTS AND HOSPITALS ENVIRONMENTS IN DIFFERENT IRAQI CITIES

The study was aimed to evaluate the prevalence of MRSA in some Iraqi hospitals and determine the most powerful methods for identification of MRSA, in order to achieve the, 278 samples were collected from different hospitals in Iraq in various intervals, 204 out of 287 were identified as Staphylococcus aureus by conventional cultural methods and microscopic characteristics and 177 isolates are identified as MRSA by using HiCrome MeReSa Agar Base medium, but 154 of 177 (87%) isolates are methicillin resistance in sensitivity test. MRSA isolates were highly resistant to β-lactam antibiotics and considered multidrug resistant (MDR) in percent of (94.9%). Touchdown PCR used to identify the isolates, 97.05% were identified as Staphylococcus aureus, while 80.88% as MRSA.

Effect of base media, FSH and anti-Müllerian hormone (AMH) alone or in combination on the growth of pig preantral follicles in vitro

To compare the efficiency of North Carolina State University medium 23 (NCSU23) and Alpha Minimum Essential Medium (α-MEM) as a base medium, and to evaluate the effects of Anti-Müllerian Hormone (AMH) alone or in combination with Follicle Stimulating Hormone (FSH) on the in vitro development and steroid production of isolated porcine preantral follicles. Porcine secondary follicles were cultured in NCSU23 or α-MEM media for 4 days. Once α-MEM was determined to be the optimal culture medium, secondary follicles were cultured in α-MEM alone or supplemented with FSH (1.5 ng/mL), AMH (50 ng/mL) or the combination of the two hormones. Follicle development was evaluated by measuring follicular growth, morphology and hormone production. There was no difference between the media NCSU23 and α-MEM in terms of follicle survival and growth (P > 0.05). However, at day 2, the antrum formation rate tended to be (P < 0.074) higher in α-MEM than NCSU23. At day 4 of culture, the estradiol and progesterone secretion were higher in α-MEM than NCSU23 (P < 0.01), while the opposite was observed for testosterone (P < 0.01). The addition of AMH and/or FSH did not affect follicular survival and growth. Nevertheless, the secretion of estradiol and progesterone induced by FSH was reduced with AMH (P < 0.01). α-MEM is a more effective base medium than NCSU23 for the in vitro follicular development of pig preantral follicles and AMH reduces the steroid production induced by FSH.

Plant Regeneration Through Somatic Embryogenesis in Two Cultivars of Pineapple (Ananas Comosus L.)

Pineapple production is mostly constrained by unavailability of high-performance suckers. However, somatic embryogenesis (SE) have been revealed the most rapid and controllable method for Pineapple propagation than conventional sucker production methods. The aim of this study was to evaluate the responses of two cultivars of pineapple regenerated through somatic embryogenesis. Calli were induced from crown leaf and plantlets leaf of Smooth Cayenne and Sugar Loaf cultivars. Murashige and Skoog base medium supplemented with vitamins B5 and different plant growth regulators combinations. BAP and / or 2,4-D have been added to base medium for calli maturation. BAP and GA3 have been added for plant elongation. The results indicated a significant influence of type of explant and copper on callus induction in pineapple cultivars. Likewise, the medium MS with NAA (0.5 mg/l) + BAP (1mg/l) has a highly significant influence with 8.8 mature somatic embryos per explant. Also, the growth regulator combinations and the cultivars have significantly influenced somatic embryos regeneration with a high rate of 55.25% plantlets using the hormonal combination BAP (3 mg/l) + GA3 (2 mg/l) for the smooth Cayenne. Leaves from plantlets constitute the explants to be used for callus induction in pineapple. The combination of BAP (1 mg/l) + copper (2 mg/l) + Picloram (6 mg/l or 12 mg/l) on Murashige and Skoog medium supplemented with vitamins B5 was favorable somatic embryos regeneration of pineapple. The protocol developed is a key study for successful mass propagation and genetics transformation of pineapple.

Simplification of culture conditions and feeder-free expansion of bovine embryonic stem cells

Bovine embryonic stem cells (bESCs) extend the lifespan of the transient pluripotent bovine inner cell mass in vitro. After years of research, derivation of stable bESCs was only recently reported. Although successful, bESC culture relies on complex culture conditions that require a custom-made base medium and mouse embryonic fibroblasts (MEF) feeders, limiting the widespread use of bESCs. We report here simplified bESC culture conditions based on replacing custom base medium with a commercially available alternative and eliminating the need for MEF feeders by using a chemically-defined substrate. bESC lines were cultured and derived using a base medium consisting of N2B27 supplements and 1% BSA (NBFR-bESCs). Newly derived bESC lines were easy to establish, simple to propagate and stable after long-term culture. These cells expressed pluripotency markers and actively proliferated for more than 35 passages while maintaining normal karyotype and the ability to differentiate into derivatives of all three germ lineages in embryoid bodies and teratomas. In addition, NBFR-bESCs grew for multiple passages in a feeder-free culture system based on vitronectin and Activin A medium supplementation while maintaining pluripotency. Simplified conditions will facilitate the use of bESCs for gene editing applications and pluripotency and lineage commitment studies.

Plant regeneration through somatic embryogenesis in two cultigroups of Pineapple (Ananas comosus L.)

BackgroundPineapple production is mostly constrained by unavailability of high-performance suckers. However, somatic embryogenesis (SE) have been revealed the most rapid and controllable method for Pineapple propagation than conventional sucker production methods. The aim of this study was to evaluate the responses of two cultivars of pineapple regenerated through somatic embryogenesis. MethodsThus, calli were induced from crown leaf and plantlets leaf of Smooth Cayenne and Sugar Loaf cultivars. Murashige and Skoog base medium supplemented with vitamins B5 and different in hormonal combinations: Auxins / Cytokins. BAP and / or 2,4-D have been added to base medium for calli maturation and BAP and GA3 for plant regeneration. ResultsThe results indicated a significant influence of type of explant and copper on callus induction in pineapple cultivars. Likewise, The medium MS with growth regulator combination NAA (0.5 mg/l) + BAP (1mg / l (BAP) has a highly significant influence with 8.8 mature somatic embryos. Also, the growth regulator combinations and the cultivars have significantly influenced somatic embryos regeneration with a high rate of 55.25% shoots by using the hormonal combination BAP (3 mg/l) + GA3 (2 mg/l) for the smooth Cayenne. Conclusion Leaves from organogenesis plantlets constitute the explants to be used for callus induction in pineapple. The combination of BAP (1 mg/l) + copper (2 mg/l) + Picloram (6 mg/l or 12 mg/l) on Murashige and Skoog medium supplemented with vitamins B5 was favorable somatic embryos regeneration of pineapple. The protocol developed is a key study for successful mass propagation and genetics transformation of pineapple.

Le sfide globali dell’era odierna da assumere come coordinate generali

In an era characterized by uncertainty and complexity efforts must be increased to define general coordinates on which to base medium-long term strategies. To this end, it is reasonable to start with the precise identification of global challenges and then define the appropriate theoretical and operational tools to face them. Indeed this chapter deals with topics concerning the following trends: 1) smart manufacturing. 2) Constraints deriving from limited basic natural resources (water, energy, food). 3) Potentialities and risks of artificial intelligence developments. 4) Changes in the workplace as a result of the increasing use of AI. 5) Suggestions on how to rethink work through two trajectories and one operational trail.

Incorporation of arginine, glutamine or leucine in culture medium accelerates in vitro activation of primordial follicles in 1-day-old mouse ovary

SummaryIn vitro activation of primordial follicles provides cancer patients subjected to oncotherapy with a safe therapeutic strategy for fertility preservation, however a successful protocol for activation of primordial follicles in prepubertal patients has not yet been defined comprehensively. There is evidence that amino acids such as leucine, arginine and glutamine could stimulate the mammalian target of rapamycin (mTOR) pathway, which plays a pivotal role in primordial follicle activation. Nevertheless, there has been no report that elucidates the effect of these amino acids on in vitro development of ovarian follicles. Therefore, the present study was conducted to evaluate the effects of these amino acids and their combination on the formation and activation of primordial follicles in 1-day-old murine ovaries during an 11-day culture period. The experimental groups consisted of base medium (BM), base medium + arginine (ARG), base medium + glutamine (GLU), base medium + leucine (LEU) and base medium + a combination of arginine, glutamine and leucine (AGL). The proportions of different stages of ovarian follicles and gene expression of regulatory factors were assessed using histology and quantitative real-time PCR on days 5 and 11 of culture. The proportion of transitional and primary follicles was greater in all amino acid-treated groups compared with the BM group (P < 0.05). Moreover, leucine resulted in elevated expression of Gdf9 and Bmp15, and glutamine augmented the expression of Pi3k on day 11 of culture. In conclusion, the present study showed that inclusion of leucine, glutamine, arginine or their combination in the culture medium for murine ovarian tissue could accelerate the activation of primordial follicles and alter the expression of the corresponding factors.

36 Extended culture after vitrification-warming helps in spindle recovery of bovine oocytes

The meiotic spindle is one of the most vulnerable cytoplasmic organelles when performing oocyte vitrification. It has been proposed that submitting oocytes to a post-warming incubation period in maturation medium helps in the reorganization of microtubules and chromosomes. Our previous experiments found no differences in spindle morphology after submitting vitrified oocytes to a 2-h incubation period. The aim of this experiment was to determine the effect of extended culture on the reorganization of the meiotic spindle of vitrified-warmed bovine oocytes. Oocytes were purchased from a commercial vendor (n=86) and matured during shipment. In this experiment, three treatments were evaluated: fresh oocytes (F) (n=30), vitrified-warmed (VW; n=26), and extended culture (EC; n=30). Cumulus-oocyte complexes were removed at 18h of maturation. Fresh oocytes were denuded by vortexing in hyaluronidase (1.5mgmL−1) and immediately fixed using 4% paraformaldehyde. Oocytes undergoing vitrification were partially denuded by pipetting in hyaluronidase (1.5mgmL−1). The vitrification protocol consisted of incubation in equilibration solution (7.5% dimethyl sulfoxide + 7.5% ethylene glycol) for 9min and then in vitrification solution (15% dimethyl sulfoxide + 15% ethylene glycol + 0.5M sucrose). While in vitrification solution, oocytes were mounted onto a Cryolock and plunged into liquid nitrogen in less than 1min. Warming was performed by placing a Cryolock into 0.5M sucrose for 3min and then into 0.25M sucrose for 3min. Finally, oocytes were washed in base medium. The base medium used for cryoprotectant and warming solutions was Dulbecco's phosphate-buffered saline supplemented with 20% fetal bovine serum. Both, vitrification and warming, were performed at 38.5°C. After warming, half of the oocytes were completely denuded and fixed and the other half underwent a 6-h incubation period in maturation medium (IVF-Bioscience). To examine microtubule distribution and chromosome arrangement, fixed oocytes were submitted to an immunostaining protocol using α tubulin antibody (1:100) and anti IgG-Alexa Fluor 488 (1:1000; Thermo Fisher Scientific) and counterstained with Hoechst. The effect of extended culture on the incidence of abnormal microtubule distribution and chromosome arrangement was analysed using logistic regression with a binomial response variable (normal/abnormal). There was no difference in maturation rates among groups (F=73.3%, VW=77%, EC=86.6%; P=0.43). For microtubule distribution, oocytes fixed immediately after warming had a higher incidence of abnormal spindles (57.7%) when compared with oocytes submitted to extended culture (26.6%; P=0.02). The most common abnormality seen in oocytes fixed after warming was small and faintly stained spindles. Microtubule distribution in fresh oocytes did not differ from oocytes in the other groups. There were no differences in chromosome arrangement among groups (P=0.11). Future research will focus on evaluating the benefits that this technique offers to improve development following IVF using vitrified-warmed oocytes.