Orc: Its Not Easy Being Green (Orc volume 1)

When rescue crosses shown in Fig. Only heteroallelic combinations of k43 chromosomes were viable. Crude embryo nuclear extracts 0 to 12 h were loaded without any additional fractionation steps on a Sephacryl S column Amersham calibrated with thyroglobulin kDa , catalase kDa , and bovine serum albumin 67 kDa as molecular mass standards. The protocols for nuclear extraction and chromatography were described previously For the salt elution experiment, nuclear pellets were extracted at the indicated salt concentration for 30 min each. Eluted proteins were fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes.

Immunoblot analysis was done by using antibodies specific for DmOrc2 and DmOrc5, horseradish peroxidase-coupled goat anti-mouse antibody as a secondary reagent Pierce and chemiluminescent detection methods. For coimmunoprecipitation experiments, extracts were precleared with protein A-Sepharose and subsequently incubated with DmOrc5-specific antibodies for 1 h in the presence of a proteinase inhibitor cocktail Complete; Roche.

1. Paracelso (Colección Lo real) (Spanish Edition).
2. Lénigme de Saint-Olav: Melchior lApothicaire – livre 1 (Une enquête de Melchior lApothicaire) (French Edition)!
3. THIS WEBSITE USES COOKIES!
4. Topolino (Excerpts From The Book Special Treatment & Other Stories 1);
5. The Kosher Marine.

Protein A-Sepharose was added for another hour. Bound complexes were recovered by sequential washes in phosphate-buffered saline Syncytial embryos between 1 and 2 h AED were collected, and embryos corresponding to Bownes' stage 4 were used for imaging. Cellularized embryos up to 6 h AED were collected, and embryos corresponding to Bownes' stages 6 to 8 were used for imaging. After manual dechorionation, embryos were mounted on a Petriperm50 culture dish Vivascience in Voltalef 10s oil Altofina.

A coverslip supplied with double-sided tape as a spacer was placed on top. For detection of GFP, the nm laser was used and the emission signal was detected at to nm.

There was a problem providing the content you requested

For detection of monomeric red fluorescent protein mRFP , a nm laser was used and emission signal was detected at to nm. Images were aquired using the time-lapse feature of the Leica confocal software. Movies were assembled either directly using this software or by using the Advanced Batch Converter Gold-Software and Quicktime. DmORC's association with mitotic chromosomes with dependence on stabilized cyclin expression was analyzed in the following genetic background: Heat shock induction was carried out essentially as described previously 38 with the following modifications: After a min heat shock, the embryos were manually dechorionated and mounted as described above.

Thus, embryos were imaged at least 30 min after the heat shock. Control experiments revealed that the conditions of the heat shock regimen did not lead to alterations in the cell cycle dynamics of DmORC data not shown. We generated a fusion of Drosophila Orc2 DmOrc2 with a fluorescent protein tag to monitor its temporal and spatial distribution in vivo. Each of these DmOrc2-GFP rescue constructs could complement the otherwise lethal combination of k43 -null alleles, when introduced in that genetic background see Fig. Therefore, by creating N-terminal as well as C-terminal fusions, which we initially pursued as a precautionary measure against potential steric constraints arising from the fusion moiety, we obtained two independent, functional DmOrc2-GFP transgenes.

The rescue strategy which we introduced here for a wild-type allele should also be applicable to the functional analysis of mutant DmOrc2 alleles, facilitating detailed analyses of initiator protein functions in vivo. Next we determined DmOrc2 protein levels in the transgenic lines by immunoblotting of embryo extracts. Upon crossing of these transgenes in a k43 -null background, the expression levels of the fusion genes were similar to that of the DmOrc2 gene in wild-type flies Fig.

Consistent with these results, we found no free, noncomplexed GFP fusion proteins when unfractionated nuclear extracts of early rescue embryos were analyzed by size exclusion chromatography Fig. To rule out this possibility, we analyzed nuclear extract from transgenic flies in two further experiments. First, when isolated embryonic nuclei were subjected to differential salt extraction, both DmOrc2 and the DmOrc2-GFP fusion protein eluted from chromatin under the same ionic conditions, with the majority of the protein extracted between mM and mM salt Fig.

Second, efficient complex integration of the fusion protein could also be monitored by coimmunoprecipitation experiments using an antibody against DmOrc5 Fig. In a further experiment, we followed the previously described decline in the expression of DmOrc genes during embryogenesis 5 , 9 , 19 , which was also evident from the abundance of maternally deposited DmOrc2-GFP Fig.

Confocal fluorescence microscopy of live rescue embryos corroborated this finding, showing a steep drop of the overall GFP signal from early to late embryonic stages. A Unfractionated nuclear extracts NE of Drosophila embryos were analyzed by S size exclusion chromatography, with subsequent immunoblot detection of DmOrc proteins. Cytoplasmic extract cyto , wash of nuclei w , and a nuclear pellet fraction after extraction np are shown as controls, as well as the total protein counterstain of the blot membrane. Immunoblot analyis of equal fractions of the nuclear extract ne at the indicated salt conditions showed that DmOrc2-GFP elutes from chromatin under the same salt conditions as the endogenous DmORC proteins.

The control shows a parallel experiment using an unrelated antibody. To suppress the signal of the immunoglobulin heavy chain Ig-hc close to the position of DmOrc5 on the blot membrane, a protein A-horseradish peroxidase conjugate was used as the secondary reagent in this immunoblot analysis D Levels of maternally deposited DmOrc proteins decrease during embryonic development. Shown is an immunoblot analysis of w wild-type wt and rescue embryos collected in the indicated time windows AED. For the rescue embryos, two live, dechorionated specimens from the indicated time windows were imaged by confocal fluorescence microscopy for GFP signal detection.

A chromosomal GFP signal appeared on anaphase chromosomes Fig. Both N- and C-terminal fusion proteins showed identical oscillatory changes in these localization studies. In the absence of any other visual marker in these GFP-only transgenic animals, it was difficult to precisely align DmOrc2-GFP loading to the course of mitotic progression. Therefore, recording more details of this process required costaining of the chromosomes. To achieve this without additional experimental interventions before and during imaging, we used a second transgenically encoded marker protein, a Drosophila histone 2A variant fused to mRFP His2AvD-mRFP His2AvD is chromatin bound throughout the cell cycle 11 , providing an omnipresent chromatin marker.

In doubly transgenic flies, the subcellular distribution and the dynamics of the GFP signal was indistinguishable from that observed in the DmOrc2-GFP rescue lines. Pictures shown in Fig. Next we asked if the dynamic shuttling of DmORC changes during embryonic development, given the major changes in cell cycle control and origin determination between the syncytium and older blastula embryos The increase in nuclear volume during interphase is paralled by a decrease in nuclear GFP signal intensity.

Preliminary quantitative evaluation of this dilution process suggests that the GFP signal, when integrated over the expanding nucleus, stays approximately constant during interphase data not shown. As expected, throughout mitosis the histone signal intensifies in more condensed chromatin. In contrast, the DmOrc2 signal is diffusely distributed in the nuclear area during early mitotic stages, with no sign of chromatin enrichment. It should be noted that in this stage of development the nuclear membrane becomes permeable to large molecules already very early in mitosis It appears that the loading process is not synchronous along chromosomes but initiates at the centromeric part and proceeds along the axis Fig.

Whatever the biological relevance, with respect to its cell cycle timing and subchromosomal localization this process is strikingly similar to the disappearance of histone H3 phosphorylation in precellular blastoderm stages of Drosophila embryos This histone modification also reflects an increase in chromatin compaction, and its reversal toward the end of mitosis could also contribute to DmORC-GFP chromosomal rebinding.

It is unclear if these findings also relate to the preferential association of Orc2 to centromeric heterochromatin found in cell lines at earlier mitotic stages 36 , 39 , as we did not see any distinct DmORC-GFP signal on metaphase chromosomes in live embryos. A Highlighted by an arrow is the interphase cell to be followed along the cell cycle.

A nuclear DmORC signal is visible. This cell cycle staging can be made as 1 min later an anaphase chromosome figure with an intense fluorescent signal becomes visible arrowheads , with part of the DmORC-GFP still dispersed C. Images in this figure are video stills taken from Movie S1 in the supplemental material. A Optical section of an embryo with asynchronously dividing cells. Nuclei in different stages of the cell cycle are marked: B Enlarged picture of a nucleus moving into metaphase.

C Enlarged picture of a nucleus moving out of metaphase. The color coding of the fluorescent proteins is as in panel A. Images in this figure are video stills taken from Movie S2 in the supplemental material. Images in this figure are video stills taken from Movie S3 in the supplemental material.

Quantification of relative fluorescence intensity along the indicated white lines is shown on top of the pictures Leica confocal software, Kernel 7. The observed binding of DmORC-GFP to late mitotic chromosomes raises the question of possible control mechanisms triggering the changes in protein localization in preparation for pre-RC assembly. We asked if loading of DmORC-GFP on chromosomes was prevented under these conditions or if the initiator protein complex followed its dynamic relocalization pattern independent of mitotic CDK activity.

Expression of the proteolysis-resistant cyclins was under the control of a heat shock promoter. Throughout the embryo, arrest in mitosis was far from complete. This is likely due to cell-to-cell variations in transgene expressivity or penetrance, in our hands precluding a conclusive analysis of potential direct changes in the phosphorylation status of DmORC subunits. In cells not affected by a mitotic cell cycle block and also in cells escaping from a transient cell cycle arrest, the DmORC-GFP signal reoccurred on the segregating late mitotic chromosomes see Movies S5 to S7 in the supplemental material , as shown before for DmOrc2-GFP transgenic flies with endogenous cyclin control.

This demonstrated that in this genetic background, control over DmORC-GFP localization was not disturbed whenever a given cell was able to progress beyond its expected mitotic cell cycle arrest point. Tragically, the proud orc clans were corrupted by the Burning Legion and used as pawns in the Legion's invasion of Azeroth. The orcs managed to rebel, however, and they were ultimately able to help turn the tide against their demon masters.

Led by the young Warchief Thrall , the orcs have reclaimed their strength and honor. The orcs moved from the Eastern Kingdoms to Kalimdor , and there they founded the nation of Durotar. Now, the orcs stand ready to fight not for the sake of conquest, but for their very right of survival in their adopted world. To their enemies, they are brutal and fearsome opponents, without parallel in their ferocity and cunning. To their allies, they are noble and honorable, following the traditions of a rediscovered past. The orcs of Lordaeron are part of a race that has once again found its true spirit, shunning the cruel practices of arcane and demonic magic for the paths of wisdom and power.

Some orcs still hang on to the arcane practices of the past, but their time is fading. Though brutal in combat, orcs fight with a feral grace that equals even the finest fencing of an elven noble. For an orc, skill in battle brings great honor. The concept of personal honor that pervades all echelons of orcish society has made the race more cohesive and more of a threat to their adversaries in the Alliance. Even the naming of an orc is temporary until he has performed a rite of passage. Once an orc has brought honor to his name and the name of his clan, the elders give him a second name based upon his deed.

For an orc, personal reputation is as important as his or her clan, and most will fight to the death in defense of either. This is in drastic contrast to when they were led by the forces tainted by the Burning Legion, and were a bestial force barely controlled by demonic magic. While the orcs might seem quick to anger, they are tempered by the wisdom of their leaders and their shamans. There are few of the demon-worshiping clans left, for the Alliance or the Horde itself has hunted down most of the remnants of such clans.

Now, most of the orcs have embraced the values embodied in the leadership of Thrall as taught by his mentor, Orgrim Doomhammer. The orcs are one of the most populous races on the world of Azeroth. While many of the Alliance races perceive them to be brutish and savage, the orcs have forged a complex society that embraces many occupations and many different races. No doubt the leadership of Thrall and the support of the tauren and jungle troll elders has aided in this solidarity, but it is quite apparent that humanity has underestimated the ability of the orcs to unify and create their own distinct and powerful culture.

Perhaps in time, the two factions might come to an understanding, but with the current division of Alliance and Horde, no one can say how or when peace might come. When they were freed from the Burning Legion, orcs experienced a cultural revolution as the unnatural, arcane bloodlust left their bodies, giving them calmer and more focused states of mind.

They, for the most part, follow Thrall as he forges ahead to reform the Horde both inside, with his shamanism, and outside, with the founding of Durotar. Still, not all orcs are pleased with these movements, namely the evil orcs who still reside on Azeroth. These orcs, who include a good number of the surviving warlocks, actively raid Alliance and Independent caravans and towns, and renounce all that Thrall proclaims the Horde is.

Like the two faces of the Alliance, the different Horde factions meet rarely; but there are some orc warlocks on Kalimdor who would very much like to retake control of the Horde forces and are gaining strength in the wilderness to do so. The orcs of Draenor had lived in a noble shamanistic society, roaming in tribes the grasslands of Nagrand on their dusty world of Draenor, for over 5, years.

They lived in peace with the draenei refuges and were at war with the ogres. Eventually, the presence of the draenei drew the Burning Legion to Draenor. After investigating their world, the powerful Eredar demon lord Kil'jaeden tricked the respected shaman Ner'zhul into striking a bargain with him. He convinced Ner'zhul that the draenei were conspiring against the orcs, and were planning on attacking. In exchange for their service to the Burning Legion , Ner'zhul and all of the orcs would receive the power necessary to conquer vast new lands, and Kil'jaeden would have an army able to crush the draenei.

To obtain this power, the orcs would need to first drink from the blood of Mannoroth the Destructor, a mighty pit lord of the Burning Legion. Grom Hellscream was one of the first to drink, and easily convinced the other chieftains and their clans to follow suit. This put them under the Blood Curse.

With the mass murder of the draenei , the elements refused to aid the orcs in their war. Believing that the elements had turned on them, the orcs turned to Ner'zhul. Gul'dan was then introduced as the new leader of the Horde. Slowly the entire race was corrupted into rampaging clans that would later be forged into the Horde. Over the next few decades the draenei and many other indigenous races of Draenor were almost utterly destroyed. Completely devoured by their demonic bloodlust and without new enemies to fight, many orc clans began fighting amongst themselves.

Petty rivalries escalated into full scale bloodbaths, and total chaos descended upon orcish society. The few remaining draenei took advantage of this and started a guerrilla campaign that continues to this day. During this time, Ner'zhul, no longer willing to watch his race destroy itself, betrayed Kil'jaeden and was subsequently replaced as Spiritual Leader of the clans by Gul'dan.

Gul'dan cared little for the Horde and easily agreed to follow Kil'jaeden in exchange for even more power.

1. Associated Data.
2. Intimate Relationships: Why They Do and Do Not Work!
3. Orc: It's Not Easy Being Green by Ed Page.
4. See a Problem?!
5. Life on Mitchell Road?
6. Is That My Child? The Brain Food Plan: Help your child reach their potential and overcome learning difficulties;
7. Books for Kids: Jungle Exploration with Captain Maverick: Adventure, habitats & Amazonian Animals (Maverick Bedtime Story Books for Kids Book 4).

Kil'jaeden taught Gul'dan how to project himself into the Twisting Nether and to commune with the dead. Gul'dan was changed by these encounters and realized how to attain even more power. Gul'dan also changed the way the orcs were ruled by giving Blackhand the title of Warchief during the war. Gul'dan gathered all warlocks who shared a desire for ultimate power and attempted to the share knowledge of communing with the dead.

Many died in the process, but the few who survived formed the initial ranks of the Shadow Council. Through careful manipulation and intricate machinations, the Shadow Council became the real ruling body of the Horde, with the clan chiefs under their thumbs. By using the promise of new lands to conquer on worlds other than Draenor, the Shadow Council was able to form a tenuous unity within the Horde.

Gul'dan and his warlocks began probing the Twisting Nether , desperately searching for new worlds within easy reach before the clans' bloodlust exploded beyond control. Gul'dan also founded the schools of Necromancy to spread the new demonic magics to even more orcs. One night, an extremely powerful entity touched the thoughts of many orc warlocks.

Gul'dan sought the advice of Kil'jaeden as to what this new presence might be, but his summons went unanswered. If Gul'dan's own tutor Kil'jaeden feared this entity, then it could prove to be a powerful tool if he could re-establish contact with it. Weeks later, Gul'dan was finally successful and opened up communication with Medivh , a sorcerer on some distant world. Gul'dan attempted to probe the designs of this Medivh, but Medivh's mind moved far too quickly for Gul'dan to discern much of any value.

Gul'dan knew with near-certainty that Medivh was attempting the same, and did not want Medivh to gain an advantage, and so quickly broke contact. Medivh later returned to the dreams of the warlocks on Draenor, presenting them with images of the land of Azeroth. The Shadow Council, despite the debate over Medivh's true intentions, decided to do Medivh's bidding if he could furnish a way to bring the horde into Azeroth. Those warlocks who were not members of the Council but had experienced the vision, were killed to keep the coming invasion secret.

Navigation menu

Weeks passed with no further word from Medivh and some members of the Council believed he was playing tricks on them. But then the rift appeared. With time and much effort the orcish warlocks were able to expand the rift enough to allow orcs to squeeze through. Though their first scouts were driven mad, either by the rift itself or by what they had seen, the council was able to confirm that on the other side of the rift was the world Medivh had shown them.

A small contingent of orcs was sent through the stabilized rift, now known as the Dark Portal , to scout and construct a base of operations. The caution urged by the Shadow Council fell on deaf ears when the clan chieftains learned of how seemingly weak the native humans of the area were. Bloodlust soon overcame the Horde, and they launched a preemptive strike against the most powerful establishment of humans in the area, the Kingdom of Stormwind. Each chieftain blamed the other for this failure, and the Horde split into two factions.

The Shadow Council attempted to reunite the Horde, but could not act directly, and so they chose an avatar to act as their puppet ruler: Blackhand the Destroyer was named Warchief of all the Horde once again. Under Blackhand's iron fist, order was restored. It was then that Medivh once again made contact with Gul'dan. Medivh seemed even more powerful, but less sane. Medivh ordered Gul'dan to have the Horde destroy the Kingdom of Stormwind, and make Medivh the new ruler of the humans.

Gul'dan initially refused to do Medivh's bidding; after all, the Horde had a new target and Medivh's usefulness, in Gul'dan's eyes, had run out. Desperate to see his plans succeed, Medivh tempted Gul'dan by promising to reveal the location of the Tomb of Sargeras , the lord of the Burning Legion and Kil'jaeden's master. And so the First War between the Horde and the humans of Azeroth occurred, ending with the destruction of the Kingdom of Stormwind. Near the beginning of that conflict the Frostwolf Clan , one of very few clans of orcs that had rejected the demonic gifts of Kil'jaeden, was exiled to Azeroth and its leader Durotan was murdered by Gul'dan's forces as a warning.

His infant son was left for dead but was taken in by a nobleman from Lordaeron fleeing the carnage of Stormwind. The Frostwolves, leaderless, fled to the far northern mountains. Toward the end of the war, a surgical strike was launched by the humans to kill the treacherous Medivh. As Medivh was assaulted, Gul'dan felt the psychic trauma waves that Medivh emanated and realized that his chance to obtain the power of Sargeras was about to slip out of his grasp. He entered Medivh's mind and attempted to steal the location of the Tomb of Sargeras while Medivh was weakened and distracted.

It was at this moment that Medivh died, and Gul'dan, having been in his mind at the time of death, was thrown into a coma. When he awoke, Gul'dan learned of a major power shift within the horde. Blackhand the Destroyer had been overthrown by Orgrim Doomhammer after he had learned of Blackhand's role in corrupting the Horde.

Doomhammer was not as gullible or easily swayed as Blackhand had been, and quickly discovered the Shadow Council's presence in orcish affairs. He completely eradicated the Council through accusations of treason. Gul'dan survived only by 'swearing' allegiance to Doomhammer, and by promising to provide a vast undead army for the Horde's use. He formed the Stormreaver Clan and began the process of re-animating the corpses of fallen knights with the spirits of the fallen members of the Shadow Council. These new Death Knights, along with other fel projects such as the capture of Alexstrasza , gave the Horde enough strength to advance steadily north despite facing the might of the unprecedented Alliance of all the human nations Lordaeron , Stromgarde , Kul Tiras , Gilneas , Alterac and the magical forces of Dalaran.

The elven nation of Quel'Thalas sent support to the Alliance, and after the Horde took their beloved lands of Khaz Modan, the dwarves and gnomes gladly joined the ranks of the Alliance. When the Kingdom of Alterac betrayed the Alliance, the victory of the Horde seemed inevitable, but the Horde was to suffer a betrayal of their own. With victory in sight, Gul'dan convinced Cho'gall of the Twilight's Hammer clan that he knew the location of the Tomb of Sargeras.

Together, along with the Stormreaver clan, they abandoned their posts and set out to claim the demonic power for their own. This loss of nearly a third of the Horde brought their campaign to a standstill at the doorstep of Lordaeron. Doomhammer, furious with the insubordination at such a critical time, deployed a large portion of his own forces to attack the deserting clans and their leaders. This allowed the Alliance forces to rally and crush the Horde while they were divided. With the destruction of the Dark Portal the Second War ended.

Privacy Policy (Updated: 5/18/18)

Although a number of powerful men in the kingdom of Lordaeron wanted the orcs rounded up and executed, King Terenas ignored them and had the orcs placed in internment camps with hopes that they would one day lose their bloodlust. There, cut off from their demonic rulers and with no way to replenish their fel stamina, the orcs languished and eventually slipped into lethargy. Even despite his vow of honor, he loved her deeply and she him. The characters were very well done and I grieved as Arianna did when she first thought him dead.

Can't wait for the next installment! A treat to read. This was a great story. I wondered if it would have enough romance in it for me, because first and foremost I'm a romance reader, but this had just the right balance for me. There is fantasy, blood magic, orcs and elves, some murder and plotting, but the heart of the story for me was the romance between a lonely princess and the orc imprisoned in a mirror in her father's castle tower. I love alien romance where the hero is not so human in appearance, and while this isn't an alien romance, it still gave me what I love in those books.

This hero is big and brutish, with grey-green skin with tattoos and scarring.

Orc (Dungeons & Dragons) - Wikipedia

He has glossy black hair and yellow eyes, not to mention tusks. This hero called to mind The Hulk: Bolthorn's a great hero. Who knew an orc could be sexy? He's protective, possessive, and very much in love. Besides his appearance, I loved most about him his sense of honor, his need for right to prevail over wrong, and his desire to protect the princess from any hurt or danger.

It all felt romantic, and I loved them getting to know each other, looking beyond appearance and seeing the true heart of a person, orc or not. There is some sex but nothing too graphic in nature. This has a lovely beauty and the beast feel to it and was just fun and romantic to read. Not overpowered by the overall story and action, the romance is complete, satisfying, and strong. The author has an easy way with her story-telling, easily catching the reader up in what's happening and making you care about the characters. The story ends with several questions to be answered; I can think of at least 10 things I need to know more about!

There is more to come in this saga, and I'll be reading the next book. Consegne illimitate in 1 giorno. Iscriviti ad Amazon Prime: Ulteriori informazioni su Amazon Prime. Metodi di pagamento Amazon. Visualizza o modifica i tuoi ordini alla pagina Il mio account. Hai bisogno di aiuto? Hai dimenticato la password? Acquista un Buono Regalo.