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stress than parents of healthy children dealing with moderately severe stressors (Barakat et al, 1997; Kazak et al, 1997) Young adult survivors, but not child or adolescent survivors, report signi cant symptoms of posttraumatic stress (Barakat et al, 1997; Rourke, Stuber, Hobbie, & Kazak, 1999; see Butler, Rizzi, & Handwerger, 1996, for an exception) Currently, interventions focus on reducing the traumatic nature of cancer and its treatment during the treatment phase in addition to providing support as families move off treatment and into the survivor phase (Kazak et al, 1999) Children with cancer must endure repeated invasive and painful medical procedures such as bone marrow aspirates, biopsies, and lumbar punctures, as these are integral components of cancer diagnosis and treatment Studies have shown that children with cancer consider painful procedures as the most unpleasant and feared aspect of cancer treatment (Fowler-Kerry, 1990).

has determinant det P(x) 1 and, hence, P(x) is unimodular, (See Section 11.3 for more details.)

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B Cell pellets (a) Grow MSCs in pellets according to Protocols 137 or 138 (b) Centrifuge tube with cell pellet at 3000 g for 10 min at 4 C (c) Aspirate supernate carefully without destroying the cell pellet (d) Add 1 ml TRIzol to each tube (e) Disintegrate the pellet with a pipette tip by pipetting 20 times up and down (f) Centrifuge tube at 12,000 g for 10 min at 4 C (g) Transfer supernate into a clear microcentrifuge tube without destroying the cell pellet, and immediately perform RNA Extraction or freeze samples at 80 C for later analysis C RNA Extraction Before starting to work, clean all working surfaces with RNAse Zap as above Change gloves frequently If not mentioned otherwise, work on ice (a) Incubate sample at room temperature for 5 min (b) Add 190 l of chloroform to each tube.

The associated pain and distress does not appear to decrease with repeated procedures and may worsen if pain is not adequately managed For example, a child who has been sensitized by a previous painful procedure that was not successfully managed may have anticipatory anxiety to the point of noncompliance or refusal to cooperate with future procedures (Miser, 1993) Our research has shown that child survivors of cancer report that getting shots and needles are among the most recalled and upsetting aspects of cancer (Kazak et al, 1996) Parents, too, report that seeing their child in pain results in fear, horror, and helplessness (Kazak et al, 1996) Thus, the optimal approach to procedure-related pain in children with cancer is critical for the well-being of the child and family and may have long-term implications for adjustment This presents a compelling clinical and research goal that requires interdisciplinary collaboration.

In clinical practice, a number of different psychological treatments are used for children undergoing painful medical procedures (eg, hypnosis, biofeedback), but the constellation of treatments commonly referred to as cognitive-behavioral therapy has been the most widely studied (eg, Chen, Joseph, & Zeltzer, 2000; Powers, 1999) In his review of literature, Powers (1999) illustrated that cognitive-behavioral interventions are a well-established treatment for reducing behavioral distress and anxiety in children undergoing painful medical procedures Typical components of a cognitive-behavioral treatment package often include breathing exercises, distraction, imagery, positive self-statements, modeling, behavioral rehearsal/role play, and positive reinforcement for using coping skills or lying still Active direction from a coach (eg, psychologist, medical staff, nurse, or parent) is another common component of CBT As these approaches are well-known and described in detail elsewhere, we have provided brief descriptions of these techniques in Table 192..

are matrices of first order partial derivatives of y(x). oy/ox' is sometimes called the Jacobian matrix of y and ay' / ox is sometimes called the gradient of y.

A clear layer should occur (c) Vortex tube vigorously for 15 s (d) Incubate sample at room temperature for 3 min (e) Centrifuge tube at 12,000 g for 15 min at 4 C An upper clear aqueous phase containing the RNA appears (f) Carefully transfer upper aqueous phase to a fresh tube (do not transfer the interface); estimate the transferred volume you can take out It is important not to disturb the lower layers during extraction this will contaminate your sample and may in uence enzyme activity of later experiments (g) Add the same volume of 70% ethanol to the homogenized lysate (estimated in the previous step), and mix well by pipetting Do not centrifuge (h) Apply up to 700 l sample, including precipitate that may have formed, to an RNeasy mini spin column sitting in a 2-ml collection tube and centrifuge at 8000 g for 15 s.

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