Analysis of Power Losses in 33/11 KV Distribution Feeder Circuits via Loss factor Technique - Method feeder 330 free download

Looking for:

Method feeder 330 free download 

Click here to DOWNLOAD

   

 

Method feeder 330 free download.Mikado Ultraviolet Method Feeder Carpfishing Rod Black | Waveinn

 

- El queria que lo guardara. Нуматака подавил смешок. Не упусти.  - Над ними склонился Фонтейн.

❿  

The 'Mixing Secrets' Free Multitrack Download Library. Method feeder 330 free download

 

To separate the two types of cells for analysis, the spheres, floating in the E8 medium, were isolated by aspirating the medium while the adherent spheres were scraped away with a cone. The well was washed twice with PBS. Colonies of polygonal cells were then dissociated with TrypLE 10 min. Culture conditions were identical to those used for culture with no feeders in E8 medium, except for EGF supplementation of the medium.

Three corneas from 3 different donors donor age ranged from 77 to 89 years were used for these experiments. A solution of 0. Following microscopic scoring after 24 hr of culture, only wells containing a single cell were used for further experiments. Cells were grown for two weeks, then enzymatically dissociated and serially subcultured at a density of 10 4 cells per well until senescence.

After microscopic scoring at day 5 of culture, only wells containing one colony were used for further experiments. Cells were grown for 12 days, then enzymatically dissociated and serially subcultured at a density of 10 4 cells per well until senescence. To promote differentiation of cultured cells into keratocytes, the cells dissociated from primary spheres cultured in E8 medium using collagenase A 0. MSCs from rat adult bone marrow were used as positive controls. Multiphoton microscopy was performed using a custom-developed laser scanning upright microscope as previously described [ 40 ].

Power at focus was 20 mW with kHz pixel rate. The 2PEF images revealed endogenous fluorescence of the cells and SHG images specifically revealed collagen fibrils without any staining [ 41 ]. Nuclei were stained with Dapi ; Molecular Probes. Twelve bit images were processed with ImageJ or FIJI, and Z-sections were projected on a single plane using maximum intensity under Z-project function.

Images were finally converted in 24 bits RGB color mode and figures were then assembled. The amount of total RNA isolated was quantified by optical density at nm. Human limbal cells from 6 donors were dissociated from superficial limbal explants, either after epithelial scraping or with no scraping. Colonies obtained in E8 medium are isolated and spheres float in the medium D.

After 2 weeks, cultured cells formed spheres with a ratio i. To evaluate the self-renewal capacity of SSC, primary spheres were dissociated and passaged under the same culture conditions. Secondary spheres were generated from dissociated primary spheres. In serial subculture, cloned cells could undergo 48 cumulative population doublings while maintaining their ability to form spheres Fig 2. After 48 population doublings, cultured cells still divided but no longer formed spheres. Isolated limbal cells from 3 donors corneas were cloned by limiting dilution.

Only wells containing 1 cell were used for culture. After 12 days, cultured cells eventually formed colonies of polygonal cells with a ratio, showing that each colony derived from a single cell. The average percentage of cells forming spheres was 1.

At confluency, each clone was dissociated and serially subcultured in the same culture conditions. Colonies were obtained at each passage until senescence characterized by cell growth arrest, which occurred after 28 population doublings Fig 2. To phenotypically characterize cells developing in spheres and colonies derived from limbal explants, we first examined the expression of various markers by immunofluorescence: markers for stem cells i. S1 Fig. In addition, expression of some of these marker genes was assessed by qPCR analysis of the spheres and colonies.

In order to promote synthesis of organized collagen fibrils, the spheres of SSC were plated on aligned nanofiber multiwell plates. Phase contrast light microscopy showed that spheres then exhibited an elongated shape parallel to nanofibers. The collagen fibrils were also orientated along the nanofibers as revealed by immunofluorescence staining and polarization-resolved SHG microscopy Fig 5B.

As shown by multiphoton microscopy, these disorganized collagen fibrils SHG signals in green are present around cells endogenous fluorescence, 2PEF in red along the full depth of spheres of SSC. B When spheres of SSC were plated on nanofiber plates to promote synthesis of an organized collagen extracellular matrix, spheres tended to elongate along nanofibers as shown by phase contrast microscopy. Aligned collagen fibrils green were also evidenced by SHG microscopy. After 14 days of culture in keratocyte differentiation medium, cells with dendritic morphology were observed Fig 6A.

When exposed to differentiating medium conditions, SSC are able to differentiate into keratocytes, myofibroblasts, fibroblasts and neurons. In contrast, cells from dissociated colonies of LSC seeded in the keratocyte and the fibroblast differentiation medium did not adhere and no cell growth was observed S2A and S2B Fig. Individual spheres obtained after primary culture of limbal cells in E8 medium, were plated in poly-D-lysine and laminin coated wells. When colonies of LSC were dissociated, and seeded in the same medium, cells did not adhere and no cell growth was observed S2C Fig.

SSC and LSC switched to adipogenic, chondrogenic or osteogenic differentiation media were assayed for mesenchymal phenotypes after 3 weeks in culture Fig 7. MSC cultured in the same conditions were used as positive controls. After primary culture, MSC used as positive controls and SSC were dissociated and plated into osteocyte, adipocyte and chondrocyte differentiating media. From small superficial limbal biopsies taken from human donor corneas, we have developed culture methods whereby both LSC and SCC could be isolated with the same medium in primary culture.

E8 medium has been used previously to grow induced pluripotent stem cells iPS and permitted the growth of iPS in feeder-free and xeno-free culture condition [ 35 , 42 ].

E8 medium is commercially available; this is of interest for cell therapy units aimed to standardize method preparation of cells for transplantation in patients. Compared with the medium described by others for the culture of SSC, E8 medium is a xeno-free medium, which does not contain FBS [ 43 , 44 ], B27 supplement [ 45 , 46 ] and cholera toxin [ 42 ]. To our knowledge, E8 medium has never been used to grow limbal epithelial and stromal stem cells. Our results show that E8 medium allows adult SSC to grow with maintenance of stemness through 48 population doublings together with various differentiation potentials.

Sixty-five to 67 cumulative populations doubling of SSC isolated from fresh bovine corneas were found by Funderburgh et al. As we used elderly donor tissues as a source of isolated limbal cells, the growth potential of our cells was probably limited, although still valuable for therapeutic developments. Results of such transplantations of elderly donor corneas are successful [ 48 ].

This issue is supported by previous studies [ 49 ]. This group found that compared with dispase-isolated sheets, LSC outgrowth generated by collagenase-isolated clusters featured larger diameter and its single cells yielded more holoclones on 3T3 fibroblast feeder layers. In the present study, after primary culture of collagenase-isolated cells, subculture of spheres resulted in growth of both adherent cells and spheres featuring SSC phenotype and no colonies of epithelial cells.

No CK3 and p63alpha positive cells were observed in these secondary cultures. In the present work, the two stem cell populations grown from limbal explants exhibited clear-cut differences regarding phenotype, growth potential and differentiation ability. Our results show that isolated human SSC expressed neural crest markers, displayed stem cell characteristics and were multipotent.

They were also able to form spheres. As opposed to SSC, LSC expressed ectoderm together with stem cell markers and were unipotent in our culture conditions. Although they originated from the same donors, the two stem cell populations showed different growth potentials.

This could be explained by the very rapid turnover of the corneal epithelium, which probably requires frequent LSC divisions. In a rabbit experimental model, the renewal time of the corneal epithelium has been shown to be 2 to 3 weeks under physiologic conditions [ 52 ]. Conversely, corneal keratocytes are thought to divide less frequently and SSC divisions are probably less frequent. Loss saving can be realized by governing the current flow through the conductors.

Voltage profile improvement occurs due to loss reduction. Traditionally, the most obtainable Reactive Volt Ampere resources in the distribution system are switchable shunt capacitor banks.

Nowadays, Distributed Energy Resources DER are another added important VAR resource because of their growing penetration, that brings innovative chances as well as encounters for loss saving. By the coordinated control of these switchable shunt capacitor banks and Distributed Energy Resources, optimum loss reduction and power quality of distribution network can be realized. Another effective technique for enhancement of voltage profile and loss reduction in distribution network is implementation of High Voltage Distribution System HVDS.

This method reduces the length of LT lines and results in loss reduction, voltage profile improvement, reduction of power failure rate, reduction of accidents, reliability etc. Also, unauthorized power theft can be avoided ,unlike in LVDS. The results show that HVDS system improves the voltage profile and minimize the loss and thus maintain power quality.

Abdullah Al-Badi. Kashem Muttaqi. Euro Asia International Journals. Puneet Chawla. Fenil Master. Anastasios Bakirtzis. IAES Journals. Anwar Sahito , Shah Murad Tunio. Hazlie Mokhlis. Ab Bin Abu Bakar. Innocent Kamwa. Seyed Mohamed Buhari. Chethan Ramesh. Mehdi Vakilian. Almoataz Y. Mani Ashouri. Gundala Srinivasarao.

Divya Teja Bonthu. The Ijes The Ijes. Ram Krishna Mishra. Sudhkara Reddy. Sivkumar Mishra. Inno Davidson. Jorge Crespo. Rakesh Ranjan , Nitin Malik. Gaddafi Sani Shehu. Mir Hadi Athari. Available in the Inline or Elasticated version. The Inline is equipped with a swivel for self strike rigging or for sliding rigging, used in combination with a Fast Change Bead. The Elasticated version allows you to use much thinner hook lengths.

For both the versions the flat bed grants a strong grip on the bottom, while the peculiar ribbed design keeps the groundbait right in position.

Method Feeder featuring an innovative shape, designed to securely hold groundbait during the cast and supplied with a matching quick release mould within every pack. The flatbed profile offers a unique grip on any bottom, while the ribbed design leaves the hookbait sitting in the dead centre, with no possibility to get the hookbait trapped on the frame, providing the perfect presentation every time, since the fed area is so confined. When fully loaded, the unique shape creates a perfect weight forward aerodynamic set up, which delivers fantastic casting accuracy and increases range.

These compact inline feeders have been designed to carry on the bottom pellets or sticky maggots, which get a quick hydration thanks to the micro holes around the body. In the commercial lakes or in slow moving waters, they attract fish downloading the hookbait in the middle of the other baits. The unobtrusive light green colour involves the lead, too, which has some 3D dots underside for better grip on the bottom.

The rubber cone, located on top of the body, gets the swivel or stops the Fast Change Bead which connects the very short hooklink to main line, while, at the same time, avoids line tangles with its elongated shape. The product is packed in a stylish blister containing two pieces, and is available in two different volumes Medium and Large.

The small size of dedicated feeders are due to the extreme conditions of use. The obtained superficial limbal rims from six donor corneas, were either scraped with a scalpel to remove the limbal epithelium three superficial limbal rims , when the aim was to obtain only stromal cells, or processed with no epithelial scraping three superficial limbal rims , when both stromal and epithelial cells were expected.

Samples were then centrifuged and the supernatant was removed. One mL of DMEM was added and samples were centrifuged a second time followed by removal of the supernatant. After centrifugation at g for 5 minutes min , the cells were resuspended in DMEM and counted with a hemocytometer.

The medium used was cholera toxin-free Green medium [ 35 ]. The culture medium was changed three times a week. E8 Medium is a xeno-free and feeder-free medium especially formulated for the growth and expansion of human pluripotent stem cells PSCs [ 37 , 38 ]. E8 medium was supplemented with 1. To separate the two types of cells for analysis, the spheres, floating in the E8 medium, were isolated by aspirating the medium while the adherent spheres were scraped away with a cone.

The well was washed twice with PBS. Colonies of polygonal cells were then dissociated with TrypLE 10 min. Culture conditions were identical to those used for culture with no feeders in E8 medium, except for EGF supplementation of the medium. Three corneas from 3 different donors donor age ranged from 77 to 89 years were used for these experiments. A solution of 0.

Following microscopic scoring after 24 hr of culture, only wells containing a single cell were used for further experiments. Cells were grown for two weeks, then enzymatically dissociated and serially subcultured at a density of 10 4 cells per well until senescence.

After microscopic scoring at day 5 of culture, only wells containing one colony were used for further experiments. Cells were grown for 12 days, then enzymatically dissociated and serially subcultured at a density of 10 4 cells per well until senescence. To promote differentiation of cultured cells into keratocytes, the cells dissociated from primary spheres cultured in E8 medium using collagenase A 0.

MSCs from rat adult bone marrow were used as positive controls. Multiphoton microscopy was performed using a custom-developed laser scanning upright microscope as previously described [ 40 ]. Power at focus was 20 mW with kHz pixel rate. The 2PEF images revealed endogenous fluorescence of the cells and SHG images specifically revealed collagen fibrils without any staining [ 41 ]. Nuclei were stained with Dapi ; Molecular Probes.

Twelve bit images were processed with ImageJ or FIJI, and Z-sections were projected on a single plane using maximum intensity under Z-project function. Images were finally converted in 24 bits RGB color mode and figures were then assembled. The amount of total RNA isolated was quantified by optical density at nm.

Human limbal cells from 6 donors were dissociated from superficial limbal explants, either after epithelial scraping or with no scraping. Colonies obtained in E8 medium are isolated and spheres float in the medium D. After 2 weeks, cultured cells formed spheres with a ratio i. To evaluate the self-renewal capacity of SSC, primary spheres were dissociated and passaged under the same culture conditions. Secondary spheres were generated from dissociated primary spheres.

In serial subculture, cloned cells could undergo 48 cumulative population doublings while maintaining their ability to form spheres Fig 2. After 48 population doublings, cultured cells still divided but no longer formed spheres. Isolated limbal cells from 3 donors corneas were cloned by limiting dilution. Only wells containing 1 cell were used for culture.

After 12 days, cultured cells eventually formed colonies of polygonal cells with a ratio, showing that each colony derived from a single cell. The average percentage of cells forming spheres was 1. At confluency, each clone was dissociated and serially subcultured in the same culture conditions.

Colonies were obtained at each passage until senescence characterized by cell growth arrest, which occurred after 28 population doublings Fig 2. To phenotypically characterize cells developing in spheres and colonies derived from limbal explants, we first examined the expression of various markers by immunofluorescence: markers for stem cells i. S1 Fig. In addition, expression of some of these marker genes was assessed by qPCR analysis of the spheres and colonies.

In order to promote synthesis of organized collagen fibrils, the spheres of SSC were plated on aligned nanofiber multiwell plates. Phase contrast light microscopy showed that spheres then exhibited an elongated shape parallel to nanofibers.

The collagen fibrils were also orientated along the nanofibers as revealed by immunofluorescence staining and polarization-resolved SHG microscopy Fig 5B. As shown by multiphoton microscopy, these disorganized collagen fibrils SHG signals in green are present around cells endogenous fluorescence, 2PEF in red along the full depth of spheres of SSC. B When spheres of SSC were plated on nanofiber plates to promote synthesis of an organized collagen extracellular matrix, spheres tended to elongate along nanofibers as shown by phase contrast microscopy.

Aligned collagen fibrils green were also evidenced by SHG microscopy. After 14 days of culture in keratocyte differentiation medium, cells with dendritic morphology were observed Fig 6A. When exposed to differentiating medium conditions, SSC are able to differentiate into keratocytes, myofibroblasts, fibroblasts and neurons.

In contrast, cells from dissociated colonies of LSC seeded in the keratocyte and the fibroblast differentiation medium did not adhere and no cell growth was observed S2A and S2B Fig. Individual spheres obtained after primary culture of limbal cells in E8 medium, were plated in poly-D-lysine and laminin coated wells. When colonies of LSC were dissociated, and seeded in the same medium, cells did not adhere and no cell growth was observed S2C Fig. SSC and LSC switched to adipogenic, chondrogenic or osteogenic differentiation media were assayed for mesenchymal phenotypes after 3 weeks in culture Fig 7.

MSC cultured in the same conditions were used as positive controls. After primary culture, MSC used as positive controls and SSC were dissociated and plated into osteocyte, adipocyte and chondrocyte differentiating media. From small superficial limbal biopsies taken from human donor corneas, we have developed culture methods whereby both LSC and SCC could be isolated with the same medium in primary culture. E8 medium has been used previously to grow induced pluripotent stem cells iPS and permitted the growth of iPS in feeder-free and xeno-free culture condition [ 35 , 42 ].

E8 medium is commercially available; this is of interest for cell therapy units aimed to standardize method preparation of cells for transplantation in patients. Compared with the medium described by others for the culture of SSC, E8 medium is a xeno-free medium, which does not contain FBS [ 43 , 44 ], B27 supplement [ 45 , 46 ] and cholera toxin [ 42 ]. To our knowledge, E8 medium has never been used to grow limbal epithelial and stromal stem cells.

Our results show that E8 medium allows adult SSC to grow with maintenance of stemness through 48 population doublings together with various differentiation potentials. Sixty-five to 67 cumulative populations doubling of SSC isolated from fresh bovine corneas were found by Funderburgh et al.

❿     ❿