The fate of the unnamed woman in Kira's bathroom. She's unable to move for fear that she may get cut by Ojirou's booby traps and his Stand could take effect.
The continuation of Johnny's life didn't end with a Happy Ending; as it turns out, he died to protect his wife from a disease. To make Johnny's life worse, when he tried to heal his wife, Rina, from her paper-folding disease with the Holy Corpse parts, he accidentally transferred the disease to his son, then to himself. The only way to make sure it didn't spread any further was kill himself. Ouch.
Josuke tears up a little when he sees a happy family playing around and is reminded that absolutely no one came to look for him when he went missing and lost his memory. The revelation that he's not even a true person due to being an amalgamation of Kira and someone else only makes it worse.
Similarly, Josuke desperately hugging Yasuho after the two were separated, because she's his anchor in 'a world of strangers'.
One scene has Holly Kira mistaking Josefumi for her son Yoshikage while the latter is in the same room, when Kira reminds her of himself, Holly realizes that the illness is making her forget about her son, and is brought to tears.
It may be worse if you take the hints of her knowing things she has no way of knowing (possibly a Stand she did not fully awaken, just like Yasuho's Paisley Park for a while). There is a chance she was actually seeing Josuke in Josefumi, and thus Yoshikage in Josuke just like she does later. Meaning that, deep down, she had an idea of what would happen with her son.
During the boat scene, Josefumi breaks down crying while Damo tortures him, saying to himself that he always gets abandoned; said internal dialogue is accompanied by a flashback to the time he drowned at the beach, showing that his mother just left him to drown.
Kira's death. While he was cold and kind of a jerk, he not only cares about his mother but has enough warmth and decency to care about others too, apologizing to Josefumi for dragging him into this. A brilliant alternate take on the villain from part 4. The worst part is that Kira dies a badass, awesome death- he dies of cardiac arrest induced from being at the epicenter of an explosion that he caused to save Josefumi's life, knowing full well he'd die. He also Dies Wide Open.
Later topped by Josefumi sacrificing his own body to save Kira via the Rokakaka fruit. And to make things worse, he clearly states that this is his death.
And the worst part is It didn't even work. The resulting fusion of Kira and Josefumi, Josuke, ended up being an entirely new being that's both and neither of them with zero memories to call his own.
Dolomite. Not only did he accidentally cause his lover's suicidal actions via awakening of his stand ability, but his attempt to rescue her turned him into the quadriplegic husk of a man we see in the manga. Not only that, but all he desires is to live alone in peace, resigned to his depressing fate. But Jobin's actions just get him wrapped up in a new mess, and tortured at the hands of Yasuho. On the plus side, he's left alone after his defeat and doesn't suffer a horrible death like the other Rock Human baddies
Yasuho nearly killed herself when she was 13 thanks to a Rock Animal tormenting her.
Rai chases down Poor Tom for the Rokakaka, and after a Traumatic Childhood Backstory monologue, is taken down in Chapter 75 by Ozon Baby after Poor Tom had a big enough opening to activate his stand to crush Rai's head in.
In Chapter 92, Holly rescues Josuke during a moment of lucidity, healing him and giving him a hint as to how to defeat the Head Doctor before she falls into a coma. Josuke breaks down crying and finally affirms that he is Josuke Higashikata and that, no matter what, he is Holly's son and calls her 'Mom'.
Chapter 95 has Jobin attacking Norisuke in front of his family, causing such severe damage that he's incapacitated. Jobin is so horrified at what he had done that he starts sobbing for his father.
Despite him being an asshole and a half, Tooru's death in Chapter 108 is still quite sad, especially the vision of a hornet that he has right before finally crumbling to dust, which turns out to be "just a memory of his."
The worst part about Tooru's death is just how... human it feels. There's no dignified final stand-off, no screaming in disbelief over his defeat. Tooru spends his last moments utterly terrified, begging for Kaato's mercy and Yasuho's help, freaking out, saying he doesn't want to die, eventually screaming in desperation and hysteria. His last words aren't even directed towards the heroes - they're a sad acknowledgment of just how alone Tooru feels, and give insight into his actual, underlying motive.
This line becomes even sadder in retrospect when you remember that Rock Humans reproduce by parasitizing hornet nests. Tooru’s last thoughts were of the closest thing to a family he’d ever known.
Kaato's death. After using the Equivalent Exchange on Tsurugi and Tooru, she's ultimately killed by bleeding out to death from an attack by Wonder of U.
The tragic thing is that her death seemed to give her the posthumous respect she didn't received from her family before. She had murdered a child, but only because the little bastard hurt her son and she didn't want to be forced to have to sacrifice her own life and be unable to raise her kids. She ended up getting caught, jailed without parole, and cut off from everyone, except Jobin, all because she wasn't willing to sacrifice her life in the first place.
The epilogue is nothing short of a Bittersweet Ending. Tooru and Wonder of U are gone, the Higashikata family's curse is removed thanks to Josuke's efforts...but Holly is still in a coma, and there's the implication she doesn't have much longer. Granted, there's a slim hope for her by Yasuho's remarks of the Locacaca Fruit might still be obtainable in their naturable habitat, but there's no guarantee Holly will live long enough for her to see the fruits of Josuke's labor.
And even if Holly does live, she'll wake up to her daughter dead and her son a completely different person.
