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The DNAzyme motor system is constructed on the AuNP. The AuNP is functionalized with hundreds of substrate strands and dozens of DNAzyme molecules that are each silenced by a locking strand. Inside the cells, the target miRNA hybridizes to the locking strand and releases the locking strand from the DNAzyme through a strand displacement reaction. The unlocked DNAzyme subsequently hybridizes to its substrate. The cofactor Mn activates the DNAzyme, which cleaves the substrate, generating two DNA segments and . The FAM-containing segment is released from the AuNP surface, restoring its fluorescence that is previously quenched by the AuNP. Meanwhile, the DNAzyme dissociates from and subsequently hybridizes to the next substrate strand, achieving the walking of the motor from one substrate strand to the next. This stepwise walking is repeated autonomously, driving the DNAzyme motor to traverse along the AuNP surface. Monitoring the fluorescence of FAM provides real-time imaging of the intracellular operation of the motor in live cells.

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Once the DNAzyme motor is taken up by the cells, the intracellular miRNA hybridizes with the locking strand through a strand-displacement reaction, releasing the locking strand from the DNAzyme. The unlocked DNAzyme then hybridizes to its substrate on the AuNP. In the presence of the cofactor Mn 2+ , the DNAzyme cleaves a substrate molecule, releasing the FAM-labelled segment. Cleavage of the DNA–RNA chimeric substrate provides the energy needed for the DNAzyme to move from one substrate strand to the next, achieving the autonomous and processive walking along the AuNP. Each walking step and substrate cleavage is accompanied by the release of the fluorescently labelled segment of the substrate. As these molecules are detached from the AuNP, they become fluorescent. Monitoring these fluorescent molecules enables real-time detection of the intracellular motion of the DNAzyme motor.

The substrate strand (sequence in Supplementary Table 1 ) is a DNA–RNA chimeric sequence composed of an RNA nucleotide flanked by two DNA domains. These two DNA domains are binding regions of two arms of the DNAzyme motor. To enhance the accessibility of the substrate strand to the DNAzyme, we added a 14-thymine (T) spacer S1 to the substrate at the 5′-end that is conjugated to the AuNP. The 3′-end of the substrate is labelled with a FAM molecule whose fluorescence is quenched by the AuNP.

The DNAzyme, a truncated form of 8–17E DNAzyme , consists of a catalytic core sequence flanked with binding Arm 1 and Arm 2 ( 1928 Jewelry 1928 Vintage Inspirations Womens Blue Y Necklace 1Tx5O5sI
). The DNAzyme is conjugated to the AuNP through a single-stranded spacer S2 linked to the 3′-end of Arm 2 . The spacer S2 comprises a 42-thymine domain that is conjugated to the AuNP and provides the spatial distance needed for the motor walking. A 16 nt domain T*1 and Arm 2 form the locking region. The sequence selection of domain T*1 depends on the specific molecules designed to initiate the motor operation. For example, to construct a DNAzyme motor that is initiated by specific intracellular miRNA, we designed a locking strand that contains a target-binding domain complementary to the target miRNA (miR-10b) and a sequestering domain complementary to Arm 2 . The hybridization of the locking strand to the domain T*1 and Arm 2 forms a duplex with a 7 nt toehold at the 3′-end of the locking strand, which sequesters Arm 2 from binding to the substrate strands on the track. With the locking strand hybridized to the DNAzyme, the DNAzyme motor is inactive. It is the inactive DNAzyme motor that is introduced to living cells and subsequently switched on by the specific cellular target. When the inactive DNAzyme motor interacts with the target molecule, for example, miRNA, the target miRNA can hybridize with the locking strand through a strand displacement reaction, exposing Arm 2 and initiating the operation of the DNAzyme motor ( Supplementary Fig. 2 ).

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A growing number of brain imaging studies in humans, along with work in rodents and non-human primates, are beginning to define the brain circuits that mediate distinct aspects of mood and emotion under normal circumstances and in various pathological conditions that are indicative of low resilience. The field has identified several limbic regions in the forebrain, which are highly inter-connected and function as a series of integrated parallel circuits that regulate emotional states ( VIDA Short Pendant Content Desires by VIDA M4TptemW
). In the sections that follow, we review how these various regions interact to mediate distinct emotional behaviours that are related to resilience. The neural regulation of endocrine and autonomic responses to stress, described in detail in REF. 70 , can be studied in humans by monitoring stress responses during functional imaging studies Novica Lapis lazuli flower earrings Blossoming Rhyme CGDZxv
.

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Figure 3
Neural circuitries of fear and reward

A simple schematic of the key limbic regions in the fear and reward circuitries. These regions are highly interconnected and function as a series of integrated parallel circuits that regulate emotional states. Each is heavily innervated by the brain's monoaminergic systems — noradrenaline (from the locus coeruleus (LC)), dopamine (from the ventral tegmental area (VTA)) and serotonin (from the raphe nuclei (not shown)) — which are thought to modulate the activity of these areas. a | Fear-inducing sensory information is relayed through the thalamus (Thal) to the amygdala (Amy). The amygdala is particularly important for conditioned aspects of learning and memory, as is best studied in fear models. The hippocampus (HP) has a crucial role in declarative memory, but it probably functions more broadly in regulating emotional, including fear, behaviour. b | The nucleus accumbens (NAc) is a key reward region that regulates an individual's responses to natural rewards and mediates the addicting actions of drugs of abuse. The prefrontal cortex (PFC) — which is composed of multiple regions (for example, the dorsolateral PFC, the medial PFC, the orbitofrontal cortex and the anterior cingulate cortex, among others) with distinct but overlapping functions — is sometimes also included in the limbic system and is essential to emotion regulation. PFC regions provide top-down control of emotional responses by acting on both the amygdala and the NAc ( a and b ). Several regions that are important for fear and reward learning are not shown in the respective circuits; for example, the NAc also regulates responses to fearful stimuli and the hippocampus also regulates responses to rewarding stimuli. The limbic regions depicted are also part of integrated circuits that mediate social responses and behaviour. The functional status of all of these circuits has important implications for resilience to stressful life events. Notably, alterations in one neurotransmitter, neuropeptide or hormone system will affect more than one circuit. Blue lines represent glutamatergic connections; green lines represent noradrenergic connections; red lines represent dopaminergic connections; the orange line represents a GABA (γ-aminobutyric acid)-ergic connection.

Neural circuitry of fear

Current models of the patho-physiology of PTSD, an example of conditions that are characterized by diminished resilience on exposure to a traumatic stressor, involve abnormal fear learning and an underlying dysfunction in the neural circuitry of fear, comprising the amygdala, the hippocampus and the ventromedial PFC (vmPFC) ( David Van Hagen Football Boot Cufflinks Silver Silver Colour tSmXcpvaMY
). Brain imaging studies in healthy participants have shown that acquisition of fear conditioning is centred in the amygdala, whereas extinction of fear memory involves both the vmPFC and the amygdala; activation in these structures, as well as the thickness of the vmPFC, has been associated with extinction success. A recent fmRI study examined the ability of physiological and neural fear responses to adapt flexibly to stimuli that changed from threatening to safe, and from safe to threatening. Both the initial fear response and the subsequent flexible shift were associated with activation of a network that includes the amygdala, the striatum and the vmPFC. In particular, the vmPFC seemed to mediate the shifting of fear to a different stimulus under stressful conditions. Findings from a recent study suggest that emotion regulation — a more advanced cognitive function — of conditioned fear might act through connections with more basic mechanisms of fear extinction, which humans share with other species.

The neural circuitry of fear is clearly important in resilience, but it has not been studied carefully in resilient individuals. It is possible that a well-functioning system in resilient individuals can prevent over-generalizing from specific conditioned stimuli, induce differential functioning of reconsolidation and extinction processes, or lead to an increased capacity for enhanced inhibition of amygdala responses by the medial PFC (mPFC) in stressful situations. Of note, preliminary findings suggest that treating patients with PTSD using cognitive behaviour therapy might have beneficial effects, by reducing amygdala activation and increasing rostral anterior cingulate cortex (ACC) activation during fear processing.

Much of our knowledge about the neural circuitry of fear comes from animal studies. The amygdala mediates the ability of cues that were associated with a fearful stimulus (for example, a footshock or a predator odour) to become aversive in their own right, as established in fear conditioning and related paradigms. The hippocampus mediates contextual and temporal aspects of fear conditioning. Reactivation of memories (that is, re-exposure to the cue or context) can lead to either reconsolidation (further strengthening of the memory) or extinction, with extinction generally requiring more intensive training. Conversely, conditioned inhibition of fear, in which animals are trained to feel protected from a threat in a certain environment, induces several antidepressant-like effects in mice. Animal studies have also made it possible to examine distinct functions of central, basolateral and medial amygdala nuclei.

Both amygdala- and hippocampus-dependent fear conditioning in animals have been related to long-term potentiation and other forms of synaptic plasticity. Accordingly, blockade of NMDA (-methyl- D -aspartate) glutamate receptors in the amygdala blocks cue-associated fear conditioning, and NMDA receptor blockade in the hippocampus blocks context-dependent fear conditioning. Consistent with the notion that extinction represents the formation of a new memory rather than the erasure of an existing memory, administration of D -cycloserine, an NMDA receptor partial agonist, can enhance extinction of fear conditioning in animal models and in patients with PTSD undergoing prolonged exposure therapy. Blockade of β-adrenergic receptors in the amygdala can also block cue-dependent fear conditioning, and β-adrenergic antagonists have been tested in patients exposed to trauma, with mixed results. In addition, fear conditioning and its extinction are regulated by activation of GRs in the hippocampus and perhaps in the amygdala, which suggests the possible use of glucocorticoids in the treatment of trauma. However, whether β-adrenergic and GR levels or functioning are different in resilient and non-resilient individuals has not yet been studied.

Neural circuitry of reward

Patients with major depressive disorder and PTSD have shown evidence of reward system dysfunction in fmRI studies, with reduced striatal activation during the performance of reward-related tasks. Altered activation of reward circuits in depressed adolescents was associated with self-reports of reduced positive affect in naturalistic settings. Differential reward system function has also been demonstrated in children of depressed versus never-depressed parents. Of note, there is evidence that inter-individual variability in neural responses to reward anticipation in healthy individuals is associated with the Val158met polymorphism.

Trait optimism, which is linked to resilience (as discussed above), might relate to reward circuit function. Sharot scanned participants who were imagining positive and negative future events. Optimism bias — the tendency to expect future events to be positive — was associated with higher activation in the amygdala and the rostral ACC when imagining positive events than when imagining negative events. The level of activation in the rostral ACC was positively correlated with dispositional optimism. Research on special forces soldiers showed that their reward-processing regions had higher reactivity than those of healthy civilian controls. Conversely, susceptibility to social-reward frustration in healthy males was associated with increased activation in prefrontal (top-down control) areas during performance of a monetary task.

Animal studies have greatly informed our understanding of the brain's reward circuitry and its possible importance for resilience. The best-established reward circuit is the mesolimbic dopamine system, which involves dopaminergic neurons of the VTA and their innervation of the nucleus accumbens and many other forebrain limbic regions ( FIG. 3 ). VTA dopamine neurons can be viewed as gauges of reward: they are activated in response to a reward (for example, food, sex or social interaction) or even the expectation of a reward, and are inhibited by an aversive stimulus or the absence of an expected reward. However, certain dopaminergic neurons are also activated by aversive stimuli, suggesting that they are more generally involved in mood regulation. Indeed, an increasing number of studies report the involvement of the VTA–nucleus accumbens circuit in depression and antidepressant responses in humans and rodents, although there is not yet a clear consensus on the role of dopamine function in resilience and vulnerability. A recent study in the social-defeat paradigm in mice (see Supplementary information S1 (box) ) has shown that increased activity of VTA dopamine neurons mediates vulnerability by increasing the activity-dependent release of BDNF onto nucleus accumbens neurons, and that resilient animals escape vulnerability in part by upregulating K channels in the VTA to prevent this increase in neuronal excitability and BDNF release ( FIG. 2 ).

Neural circuitry of emotion regulation

A greater capacity for emotion regulation has also been related to stress resilience, and studies of individuals with psychiatric disorders suggest that they have abnormalities in their emotion regulation systems. A neural model of emotion regulation consisting of ventral and dorsal systems has been described, with various patterns of abnormalities associated with a range of psychiatric disorders. Studies in mood and anxiety disorders have most consistently identified abnormalities in amygdala, hippocampus, subgenual ACC and PFC function.

In healthy individuals, differential amygdala reactivity to negative stimuli could represent an intermediate phenotype associated with vulnerability to anxiety and depressive disorders. Indeed, several studies have linked the short allele of and the Met158 allele of with higher anxiety levels, vulnerability to negative mood, increased amygdala reactivity to negative stimuli and altered functional coupling between the amygdala and the cortex. Furthermore, individual differences in cortico-limbic connectivity suggest that some people might have a genetic predisposition to inflexible emotion processing. As mentioned above, recent imaging studies have shown evidence that multiple gene interactions have an effect on limbic reactivity to unpleasant stimuli. In addition, studies of healthy children and young adults at high familial risk for depression have yielded evidence that their neural responses to emotional stimuli differed from those of controls at low familial risk for depression.

One mechanism of emotion regulation — cognitive reappraisal — has received particular attention. fMRI studies have shown increased activation in lateral and medial PFC regions and decreased amygdala activation during reappraisal, with increased activation in the lateral PFC associated with reappraisal success. It has thus been suggested that the PFC regulates the intensity of emotional responses by modulating the activation of the amygdala.

A recent fMRI study using mediation analysis demonstrated that the vlPFC acts on both the amygdala and the nucleus accumbens, resulting in opposite behavioural responses: the pathways through the amygdala and the nucleus accumbens were associated with reduced and increased reappraisal success, respectively. These findings are consistent with animal studies, which have established that the amygdala and the nucleus accumbens work in concert to regulate an individual's responses to both negative and positive emotional stimuli. Thus, variability in the functions of these two pathways might underlie individual differences in emotional response and emotion regulation in stressful contexts. Greater use of reappraisal in everyday life has also been linked to greater PFC and lower amygdala activation to negative stimuli, suggesting that there might be a central mechanism through which reappraisal could promote successful coping and reduce the risk of mood disorder onset.

A recent fMRI study found that resilient women with a history of sexual trauma were more successful at cognitively enhancing emotional responses to aversive pictures than women with PTSD after sexual trauma and healthy, non-traumatized controls. This increased capacity to enhance emotional responses was associated with increased PFC activation. These results highlight the complexity of emotion regulation systems and suggest that resilience could also be associated with the ability to sustain attention to unpleasant stimuli. Perhaps this increased attention is related to a more accurate or optimistic appraisal of the perceived threats.

Additional neural circuits relevant to social behaviour

The capacity for empathy enables individuals to generate appropriate emotional responses in social contexts and might be related to social competence, which is a characteristic of resilient individuals. Recent years have seen a surge of interest in the study of the so-called mirror neuron system, which comprises cortical neurons that fire similarly when an animal performs a task or observes another animal of the same species performing that task. It is proposed that this system, acting in conjunction with limbic brain regions, has a central role in understanding others' emotions and intentions. In humans, the vmPFC is activated both when people think about their own mental states and when they think about those of other people, and patients with lesions of this region have deficits in social emotions such as shame, guilt and empathy. Much future work is needed to understand possible links between the capacity for empathy, mirror neuron system function and resilience. Preliminary findings of greater activation in presumed mirror neuron and associated limbic areas during imitation of emotional faces in children with higher levels of empathy and interpersonal competence are promising. Of note, a recent study suggests that the neuropeptide oxytocin could improve a person's ability to infer the mental states of others.

The role of oxytocin in promoting social attachment in humans has recently received increased attention. A study in healthy men participating in a laboratory-based economic trust game showed that intranasal administration of oxytocin increased trust, and suggested involvement of the amygdala. Imaging studies have demonstrated that mutual cooperation induces activation in reward circuitry regions, which are modulated by oxytocin and vasopressin. Conversely, oxytocin reduced amygdala activation in response to fear-inducing visual stimuli and reduced connectivity between the amygdala and brainstem areas that mediate autonomic and behavioural fear responses. Oxytocin thus seems to facilitate social attachment by enhancing the reward value of social stimuli and reducing potential fear responses. In laboratory animals, central release of oxytocin and vasopressin regulates anxiety and social behaviour. In rodent species, oxytocin and vasopressin increase social recognition, pair bonding and affiliation.

Social contact promotes health and well-being. An fMRI study of married women demonstrated that holding hands with their husband attenuated neural responses to the threat of receiving a shock, a response that was proportional to the quality of their relationship. As discussed above, social competence and openness to social support are core characteristics of resilient individuals, and these qualities might help to modulate central responses to stress in these individuals. The effects of social contact on neural responses to threat, and the potential involvement of neuropeptides that promote social attachment, warrants direct investigation in resilient individuals.

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Stress resilience refers to an individual's capacity for successful adaptation to acute stress, trauma or more chronic forms of adversity. Although the range of complex mechanisms that lead to resilient phenotypes is far from being fully determined, a model of resilience has begun to emerge from the study of adaptive stress responses at multiple phenotypic levels. Beginning in development, an individual's genes and their inter action with environmental factors (and perhaps with stochastic epigenetic events) shape the neural circuitry and neuro chemical function that are expressed in an observable range of psychological strengths and behaviours characteristic of resilient individuals. Various genetic polymorphisms affect a person's limbic reactivity and prefrontal-limbic connectivity, influencing their initial responses to negative or traumatic events, as well their capacity for cognitive reappraisal of those events.

Ten years ago, as downtown was cordoned off for security and recovery purposes, many people thought it would take the city decades to recover. There were predictions that businesses would flee, residents would leave for the suburbs and crime would return.

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We bonded in grief with neighbors and strangers -- and we vowed to bring our city back, in defiance of the terrorists and in honor of the victims.

Each of us did that in our own ways: by flying flags and supporting our first responders. By volunteering our time and donating our blood. By supporting families who lost loved ones. And, most of all, by living our lives without bowing to fear. Together, and with the generous support we received from people across the country and around the world, we began bringing our city back, faster and stronger than anyone thought possible.

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, Dallas and Philadelphia -- combined. But those numbers don’t begin to tell the story of the transformation that has occurred downtown.

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Before the attacks, lower Manhattan had been a business district, but not much of a neighborhood. In 2002, our administration outlined a new vision for lower Manhattan. We believed that we could create a vibrant and dynamic 24/7 neighborhood by investing in the parks, schools and cultural facilities that attract families -- and the infrastructure that attracts commercial development.

Today, with essential support from the state and federal governments, that vision has come to life.

Now, when you walk around downtown, you’ll see streets bustling not just with bankers, but with baby strollers. You'l see new restaurants, new bars, new shops, new housing, new schools -- with 4,000 more classroom seats. And you’ll see new parks and playgrounds, many of them along the waterfront. For instance, take Pier 25 on the Novica Rhodium plated amethyst dangle earrings Pretty Lilac UtcDPE
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Weddings

By The New York Times

165 Years of Love (and War) in The New York Times Wedding Announcements

Portraits of a man and woman believed to be Sarah Mullett and John Grant, depicted around 1851, the year of their joyous union. Christopher Capozziello for The New York Times

By Lois Smith Brady

The debut issue of The New York Times on Sept. 18, 1851, known then as David Yurman Osetra Medium Rhodolite Garnet Hoop Earrings fLWD7
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, and one wedding announcement only, for the newlyweds Sarah Mullett and John Grant. He was 26 and she was 23. The ceremony took place at the Trinity Episcopal Church in Fredonia, the town in western New York where the bride had grown up. The bridegroom was from Jamestown. They were married Sept. 10 (though from the smudgy looks of the article, the Times reporter got it wrong and reported Sept. 15), by the Rev. T. P. Tyler. The announcement was a mere sentence, a bare-bones beginning for a Times institution.

So why were John and Sarah the chosen ones?

Based on old newspaper clippings and census reports, it appears that the Mulletts were a prominent family in Fredonia. The bride’s father, James, epitomized upward mobility. He grew up on a farm in Vermont, the oldest of 13 children, and made his way to Fredonia, where he became a self-taught lawyer (such a thing was possible in those days). He eventually became a State Supreme Court justice.

The bridegroom was a cousin of Ulysses S. Grant, then an Army lieutenant, who would go on to become a general and president.

Today, oil portraits of John and Sarah hang side by side in the New Haven family room of Wendy Grant Haskell, 63, who is a psychotherapist and the Grants’ great-great-granddaughter. Each is wearing black, and a similarly dour and humorless expression, as if they had whispered to each other beforehand, “Don’t smile!”

“I think they are trying to project importance, status and prestige,” Ms. Haskell said, adding that the portraits were probably painted soon after the wedding.

(When I first called Ms. Haskell to inquire about John and Sarah Grant, she was flummoxed; I think she thought I was trying to sell her a Times subscription. When I explained my reason for calling, she said she had no idea the Grants had appeared in the newspaper’s first wedding announcement. “What a hoot!”)

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Her brother David Marshall Grant, an actor you might recognize from “The Devil Wears Prada" or his Tony-nominated performance as Joe Pitt in “Angels in America,” said: “I would guess, looking at those portraits, that it was a less joyful time. I’m sure there were struggles just to get by. I can’t imagine what it was like just to make breakfast. I’m angry when there’s a line at Starbucks.”

John and Sarah Grant’s lives probably did not turn out exactly as they envisioned. After the wedding, they settled in Jamestown, where John was co-owner of a grocery store dealing in tea, fruit, molasses and salt, among other products. Sarah kept house, according to the census, and had a live-in Irish servant. The couple had one son, James Mullett Grant, who married and also had one son, Havens Grant. Then, sometime before the turn of the century, James Mullett Grant traveled alone to Havana and never returned.

“John and Sarah Grant were all about the promise of being a really significant couple starting a whole line of prosperity, but then look at what happened,” Ms. Haskell said. “Their son fell off the edge of the earth.”

On June 2, 1883, John Grant’s obituary appeared in The Evening Observer, a local newspaper. As brief and terse as his wedding announcement, the notice merely reported that he had “dropped dead” in his home at age 60. Sarah Grant died 29 years later.

Then things got even more interesting.

The couple’s grandson, Havens Grant, went to Yale, became a lawyer in New York City, married a free-spirited poet and ran a moonshine business on the side during Prohibition.

Today, the Grant descendants gather every Christmas at Ms. Haskell’s home in Connecticut and take a family photo with John and Sarah’s portraits in the background. In a recent photograph, everyone is wearing colorful tie-dyed scarves or shirts. David Marshall Grant appears with his husband, KC Reischerl, and their daughter, Evelyn May Reischerl Grant, who is 6 and the youngest descendant. She is also the only female family member in the photo. Ms. Haskell’s three sons are gay and appear with their partners.

Ms. Haskell’s son David Haskell is an editor at New York magazine and is carrying on the family moonshine tradition. He is a co-owner of Kings County Distillery , which is based in Brooklyn Navy Yard and makes craft whiskeys.

What would John and Sarah Grant think? If they were to see the lineup of gay couples in tie-dye, David Marshall Grant said, “I think they would think they had gone to Mars; it’s an astonishing example of what happens in time.”

See the announcement in the first issue of The New-York Daily Times.

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