From marrow to brain: expression of neuronal phenotypes in adult mice.

After intravascular delivery of genetically marked adult mouse bone marrow into lethally irradiated normal adult hosts, donor-derived cells expressing neuronal proteins (neuronal phenotypes) developed in the central nervous system. Flow cytometry revealed a population of donor-derived cells in the brain with characteristics distinct from bone marrow. Confocal microscopy of individual cells showed that hundreds of marrow-derived cells in brain sections expressed gene products typical of neurons (NeuN, 200-kilodalton neurofilament, and class III beta-tubulin) and were able to activate the transcription factor cAMP response element-binding protein (CREB). The generation of neuronal phenotypes in the adult brain 1 to 6 months after an adult bone marrow transplant demonstrates a remarkable plasticity of adult tissues with potential clinical applications.

Prenatal organochlorine exposure and measures of behavior in infancy using the Neonatal Behavioral Assessment Scale (NBAS).

BACKGROUND: Previous literature suggests an association between organochlorines and behavioral measures in childhood, including inattention. OBJECTIVE: This study was designed to assess whether prenatal organochlorine exposure is associated with measures of attention in early infancy. METHODS: We investigated an association between cord serum polychlorinated biphenyls (PCBs) and p,p'-dichlorodiphenyl dichloroethene (DDE) levels and measures of attention from the Neonatal Behavioral Assessment Scale (NBAS) in a cohort of 788 infants born 1993-1998 to mothers residing near a PCB-contaminated harbor and Superfund site in New Bedford, Massachusetts. RESULTS: Medians (ranges) for the sum of four prevalent PCB congeners and DDE levels were 0.19 (0.01-4.41) and 0.30 (0-10.29) ng/g serum, respectively. For the 542 subjects with an NBAS exam at 2 weeks, we observed consistent inverse associations between cord serum PCB and DDE levels and NBAS measures of alertness, quality of alert responsiveness, cost of attention, and other potential attention-associated measures including self-quieting and motor maturity. For example, the decrement in quality of alert responsiveness score was -0.51 (95% confidence interval, -0.99 to -0.03) for the highest quartile of exposure to the sum of four prevalent PCB congeners compared with the lowest quartile. We found little evidence for an association with infant orientation, habituation, and regulation of state, assessed as summary cluster measures. CONCLUSIONS: Our findings provide evidence for an association between low-level prenatal PCB and DDE exposures and poor attention in early infancy. Further analyses will focus on whether organochlorine-associated decrements in attention and attention-related skills in infancy persist in later childhood.

Molecular mechanisms of action of new xenobiotic immunosuppressive drugs: tacrolimus (FK506), sirolimus (rapamycin), mycophenolate mofetil and leflunomide.

Among all the new immunosuppressive molecules being investigated either preclinically or clinically, four stand out: tacrolimus (FK506), sirolimus (rapamycin), mycophenolate mofetil and leflunomide (and its malononitriloamide analogs). Each drug has distinct mechanisms of immunosuppressive action, and in the past year significant advances have been made in our understanding of the actions of these drugs at the molecular and even atomic levels. Data from recent clinical trials demonstrate that these drugs very effectively suppress graft rejection or autoimmune diseases, validating the pivotal role played by each of their distinct molecular targets in the normal functioning of immune cells.

The role of respiratory epithelium in a rat model of obliterative airway disease.

BACKGROUND: The etiology and pathogenesis of obliterative bronchiolitis after lung transplantation remain to be fully elucidated. Using a rat model of heterotopically transplanted trachea grafts, we have examined the role airway epithelium plays in obliterative airway disease (OAD). METHODS: Rat trachea isografts were denuded of epithelium by protease digestion. Grafts were inoculated either with or without native airway epithelial cells and transplanted into the omentum of recipient animals. RESULTS: Airway epithelium removal resulted in OAD in denuded isogeneic trachea grafts. Reseeding of the denuded grafts with epithelial cells significantly reduced airway obliteration from 78% to 22% luminal occlusion. CONCLUSIONS: Non-immune-mediated injury will cause OAD, and epithelial cell replacement in denuded isografts can significantly reduce the fibrotic progression of the disease.

Prolonged inhibition of obliterative airway disease in murine tracheal allografts by brief treatment with anti-leukocyte function-associated antigen-1 (CD11a) monoclonal antibody.

BACKGROUND: We have previously shown that anti-leukocyte function-associated antigen (LFA)-1 (CD11a) monoclonal antibody (mAb) prevents acute rejection and produces donor-specific unresponsiveness in murine recipients of heterotopic heart allografts. Here, we investigate the ability of this mAb to prevent the development of obliterative airway disease (OAD) in murine recipients of tracheal allografts. METHODS AND RESULTS: BALB/c tracheae were heterotopically transplanted into C3H mice. OAD developed by day 28 after transplantation and was characterized histologically by a loss of epithelial cell coverage and luminal obliteration of the tracheal allograft with a proliferation of fibrogenic mesenchymal cells, which is a lesion comparable to obliterative bronchiolitis in human lung transplant recipients. Monotherapy with anti-LFA-1 mAb preserved graft epithelium, prevented the development of OAD, and maintained unresponsiveness to donor antigen for more than 42 days after the final mAb administration. CONCLUSION: These findings suggest the potential for anti-LFA-1 mAb therapy to suppress both acute and chronic rejection in clinical lung transplantation.

Immunosuppressive therapies for the prevention and treatment of obliterative airway disease in heterotopic rat trachea allografts.

BACKGROUND: Obliterative bronchiolitis remains a major long-term complication after lung transplantation. Using a reproducible model of heterotopically transplanted rat tracheas, this study examined the role of several novel immunosuppresive compounds to prevent and reverse obliterative airway disease in these animals. METHODS: Brown Norway rat trachea were transplanted into the greater omentum of Lewis (allografts) or Brown Norway (isografts) animals. Recipient animals were treated with rapamycin, cyclosporine, 15-deoxyspergulin, mycophenolate mofetil, or leflunomide from day 0, 7, or 14 until day of graft removal, either day 28 or 50. Trachea segments were evaluated for degree of lumenal occlusion, as well as percent and type of lumen epithelial cell coverage. RESULTS: All untreated allografted tracheas obliterated completely, although isografts appeared patent with normal respiratory epithelium when they were removed. Leflunomide, rapamycin, and cyclosporine effectively prevented obliteration when treatment was initiated at day 0, with rapamycin showing continued efficacy when initiated as late as day 7. 15-deoxyspergulin and mycophenolate mofetil failed to consistently inhibit obliteration with any treatment schedule. An inverse correlation was found between epithelial coverage and degree of obliteration, and was especially pronounced in grafts from cyclosporine-treated animals. CONCLUSIONS: Immunosuppressive drug therapy will inhibit airway obliteration, but efficacy sharply diminishes if initiation of treatment is delayed. Efficacy also varies among immunosuppressive compounds, and results indicate those drugs that enable epithelial regrowth most effectively inhibit airway graft obliteration.

Epithelial re-growth is associated with inhibition of obliterative airway disease in orthotopic tracheal allografts in non-immunosuppressed rats.

BACKGROUND: Because epithelial cells are targets of alloimmune injury leading ultimately to airway obliteration, we tested whether epithelial re-growth could prevent obliterative airway disease (OAD) in orthotopic tracheal allografts. METHODS: Brown Norway tracheal segments were orthotopically transplanted into nonimmunosuppressed Lewis rats. Allografts were removed on days 2-10 (n=13), 30 (n=4), and 60 (n=5) for histology, computerized morphometry (obliteration), and immunohistochemical detection of mononuclear cells, smooth muscle alpha-actin, and tissue phenotype. Normal tracheas, host tracheas, and heterotopically transplanted allografts served as controls. RESULTS: Orthotopic allografts removed on days 2-10 exhibited epithelial damage and re-growth and mononuclear cell infiltration. On days 30 and 60, partially ciliated cuboidal or attenuated epithelium completely covered the lumen. Although mononuclear cells declined, numerous T cells with a high CD4/CD8 ratio were found in the epithelium till day 60. Orthotopic allograft epithelium expressed donor phenotype on day 7, but recipient phenotype on days 30 and 60. Despite subepithelial alpha-actin positive myofibroblast proliferation, obliteration did not progress from day 7 to 30 and 60 (35, 30, and 33%, respectively). Although more than in normal or host tracheas, the obliteration in orthotopic allografts on days 30 and 60 was significantly less (P<0.001) than in heterotopic allografts. CONCLUSIONS: We describe, for the first time, longterm patency of fully histoincompatible orthotopic tracheal allografts in nonimmunosuppressed rats. Despite acute alloimmune injury and induction of myofibroblast proliferation, epithelial re-growth from the host limited the progression of OAD, thus emphasizing the role of epithelium in the control of airway obliteration.

Obliterative airway disease after heterotopic tracheal xenotransplantation: pathogenesis and prevention using new immunosuppressive agents.

BACKGROUND: The purpose of this study was to investigate whether obliterative bronchiolitis might occur after xenogenic pulmonary transplantation. A model for obliterative airway disease (OAD) after tracheal allograft transplantation in the rat undergoes tracheal obliteration with histologic features characteristic of obliterative bronchiolitis in human lung transplant recipients. Using this model, the pathogenesis of OAD and its prevention with immunosuppressive drugs was studied in rat recipients of hamster tracheal grafts. METHODS: Tracheae from 30 hamsters (xenografts) or 23 Brown-Norway rats (allografts) were implanted and wrapped in the greater omentum of untreated Lewis rats. The grafts were removed on day 1, 3, 7, 14, 21, or 28 after transplantation and stained with hematoxylin and eosin and Masson's trichrome and by immunohistochemistry and immunofluorescence (IFL) techniques. In addition, 25 recipients were treated with cyclosporine (CsA, 10 mg/kg p.o.), leflunomide (LFM, 20 mg/kg p.o.), or rapamycin (RPM, 6 mg/kg i.p.) for 14 or 21 days (5 animals per treatment group). Visual and morphometric analyses were used to evaluate the extent of airway obliteration, luminal coverage by respiratory or flattened cuboidal epithelium, and extent and density of peritracheal cellular inflammation. RESULTS: In all xenografts, a neutrophilic infiltration of the mucosa and submucosa was observed from day 1 until day 14 and was associated with complete loss of tracheal epithelium by day 14. A marked peritracheal mononuclear cellular infiltrate mixed with plasma cells and eosinophils was seen on days 7 and 14. Both the extent of peritracheal inflammation and the density of the mononuclear cell infiltrate were significantly increased in xenograft tracheae when compared with the allografts. Tracheal obliteration began on day 14 and reached a maximum of 43% on day 21 with evidence of intraluminal fibrosis. In contrast to IFL of allografts, IFL of xenografts demonstrated marked deposition of rat immunoglobulin in the peritracheal tissue on days 7 and 14. The effects of treatment with immunosuppressive drugs on tracheal graft narrowing and protection of respiratory epithelium were as follows: After 14 days of treatment, the percentage of tracheal graft narrowing was 12%, 23%, and 19% in the no treatment, CsA, and LFM groups, respectively; the percentage of respiratory epithelium at 14 days was 0%, 21%, and 95%. After 21 days of treatment, the percentage of tracheal graft narrowing was 43%, 49%, 12%, and 5% for the no treatment, CsA, LFM, and RPM groups, respectively; the percentage of respiratory epithelium at 21 days was 0%, 39%, 86%, and 0%. Using computerized morphometry, the extent and densities of the peritracheal cellular infiltrates were significantly reduced in LFM- and CsA-treated groups when compared with untreated xenograft controls. LFM and RPM, but not CsA, significantly reduced the degree of luminal obliteration compared with no treatment (P<0.05). LFM and, to a lesser extent, CsA were able to prevent the loss of normal respiratory epithelium. Analysis by IFL revealed a marked decrease in rat immunoglobulin deposition in xenografts from LFM- and RPM-treated groups compared with xenografts from CsA-treated or untreated rats. CONCLUSIONS: (1) OAD occurs not only after tracheal allotransplantation but also after xenotransplantation. (2) Subepithelial infiltration of neutrophils and the appearance of plasma cells and eosinophils in the peritracheal infiltrates distinguished the histology of rejected xenografts from allografts. (3) Antibody deposition was detected by IFL only in xenografts. (4) Treatment with LFM or RPM significantly decreased the severity of luminal obliteration. Importantly, LFM also prevented the loss of respiratory epithelium.

A comparative study of the behavior of Greek neonates.

Three groups of 30 Greek neonates each (an adoptive group from the Metera orphanage, a lower-class group, and a middle-class group) were evaluated at days 1, 5, and 10 after birth using a behavioral scale of 21 items and a neurologic evaluation of 16 items. Behaviors were examined for group differences and group-by-age recovery curves were determined during the first ten days. Significant differences were found in the separate items and items grouped to reflect interactive, motor, and state behavioral dimensions. The adoptive babies at the Metera orphanage generally performed the most poorly. This difference seems not only to reflect intrauterine differences, especially in regard to nutrition, but to point to the likelihood of eliciting less than optimal responses from future environments. The middle-class group had the worst scores on physiologic items and were similar to the Metera babies in having initially depressed interactive, motor, and state behavior. Improvement in these areas over ten days suggested that temporary effects of maternal medication caused the poor scores. The recovery curves of the infants pointed to the important effects of such perinatal variables as maternal medication on early neonatal behavior.

Regional obstetric anesthesia and newborn behavior: effect over the first ten days of life.

This study examined the effects of carefully controlled amounts of analgesic premedications and anesthetics administered to mothers during delivery on the behavior of the newborn over the first ten days of life. The subjects were selected to minimize the synergistic effects of medication and other stress factors, such as abnormalities of pregnancy, labor, or delivery. The effects of these drugs on the behavior of these infants was small. The data provide a picture of the behavioral recovery of a group of minimally stressed newborns.

Enhancement of obliterative airway disease in rat tracheal allografts infected with recombinant rat cytomegalovirus.

BACKGROUND: Cytomegalovirus infection has been identified as a significant risk factor for the development of obliterative bronchiolitis in human lung transplant recipients. This study was designed to assess the influence of rat cytomegalovirus (RCMV) on the pathogenesis and development of obliterative bronchiolitis in an experimental model of obliterative airway disease, which occurs after allogenic heterotopic tracheal transplantation in rodents. METHODS: Sixty Lewis rats were infected intraperitoneally with 10(7) plaque-forming units of recombinant lac-Z-tagged RCMV expressing the gene for beta-galactosidase. Rats were either infected at the time of surgery (acute infection, n = 30) or 56 days before surgery (chronic infection, n = 30). Tracheae from Brown Norway (allograft) or Lewis (isograft) rats were implanted and wrapped in the greater omentum of infected Lewis rats. RCMV infection was verified in different recipient tissues by in vitro plaque-assays and by direct in situ staining for beta-galactosidase activity. The tracheal grafts were harvested on days 7, 14, and 21 after transplantation and stained with hematoxylin-eosin and Masson's trichrome. The peritracheal cellular inflammation was scored visually. The cellular density of the infiltrating cells and the extent of airway obliteration were analyzed by use of computer-digitized morphometry and compared with uninfected allografts as control. RESULTS: Both acute and chronic cytomegalovirus infection produced significantly higher mononuclear cell density values on days 7 and 14 compared with noninfected controls, indicating a more intense immune response in the infected allografts. Tracheal allograft obliteration was also more extensive after acute and, in particular, after chronic cytomegalovirus infection (64% narrowing after 21 days compared with 36% in grafts from noninfected control animals). CONCLUSIONS: Our experimental results provide direct evidence that the tracheal grafts were infected with RCMV and that the development of obliterative airway disease was enhanced in the acutely and chronically infected allografts compared with grafts from noninfected control animals.

RAD in stable lung and heart/lung transplant recipients: safety, tolerability, pharmacokinetics, and impact of cystic fibrosis.

BACKGROUND: RAD is a novel macrolide with potent immunosuppressive and antiproliferative activities. This study characterizes the safety, tolerability, and pharmacokinetics of two different single oral doses of RAD in stable lung and heart/lung transplant recipients with and without cystic fibrosis (CF). METHODS: This was a Phase I, multicenter, randomized, double-blind, two-period, two-sequence, crossover study. Single doses of RAD capsules at doses of 0.035 mg/kg (2.5 mg maximum) or 0.10 mg/kg (7.5 mg maximum) were administered with cyclosporine (Neoral [cyclosporine, USP] modified), steroids, and azathioprine on Day 1. The alternate dose was administered on Day 16. Laboratory assessments, vital signs, and adverse events were recorded throughout the study. RAD pharmacokinetic profiles were assessed over a 7-day period following each dose. Steady-state cyclosporine (CsA) profiles were assessed at baseline and with each RAD dose; RAD and CsA trough concentrations were obtained throughout the study period. RESULTS: Of the 20 patients randomized, 8 had CF and 12 did not. Single doses of RAD were safe and well tolerated. Headache was the most common side effect. RAD produced a mild, dose-dependent, reversible decrease in platelet and leukocyte counts. Cholesterol and triglycerides were minimally affected. At both doses, CF patients had significantly lower peak concentrations of RAD than did non-CF patients (p = 0.03); however, overall exposure (area under the curve/dose) was not different between the groups (p = 0.63). At the higher dose, there was a clinically minor under-proportionality in AUC, averaging -11%. Steady-state pharmacokinetics of CsA were not affected by RAD co-administration.RAD was safe and well tolerated by stable lung and heart/lung transplant recipients with and without CF. The presence of CF did not influence the extent of RAD exposure. Single doses of RAD did not affect the pharmacokinetics of CsA. Ongoing studies are assessing the long-term safety and efficacy of RAD in lung and heart/lung transplantation.

Working with families. Opportunities for early intervention.

The birth or arrival of a baby is a defining event for families. Most infants progress through predictable, yet individual, patterns of development. All families want the opportunity to optimize their children's futures, yet each spurt of development is a transition that can stress the child and the entire family system. These biobehavioral shifts, or "touchpoints," provide opportunities for the pediatrician to help the family understand, adapt, enhance, and promote the child's outcome. This family level approach serves as a form of anticipatory guidance that helps both parent and child successfully manage the challenging phases of development: motor, cognitive, social, and emotional. The "touchpoints" concept can be adapted for the pediatrician to help older children and their families.

The infant at risk.

The opportunity in early infancy for joining the developmental processes of infants with the energy in the parents for bringing the infants to their best potential has been out of proportion to our expectations. By visualizing the capacity of babies to organize around the positive experiences of interaction with a nurturing adult in our neonatal assessments, we have better understood normal recovery processes from labor and delivery and plasticity for recovery from CNS insults in the neonate. As we observed and identified with them to produce best behavior in the infants, their parents were able to believe in our methods and to work toward their infants' optimal recovery. Hence, our assessments of premature and normal infants became a window for us and for parents into organization and ongoing development. In later infancy, the face-to-face procedure served a similar purpose. In other words, within the observational assessment of a small baby is contained not only the organizational capacities of his or her present status, but also an opportunity to see what he will do to his parents and what they will have to do to organize him. By sharing this observation with them, we share these processes and give them an opportunity to identify the baby's positive potential in addition to his or her deficits. As such, parents can utilize the forces for nurturance in their relationship to produce the baby's optimal recovery. Serial observations over time provide an opportunity to observe the baby's capacity to organize himself as he recovers from a known set of experiences surrounding labor, delivery, and a new environment.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of kangaroo care and standard care: behavioral organization, development, and temperament in healthy, low-birth-weight infants through 1 year.

OBJECTIVE: To determine whether Kangaroo Care (KC) for healthy, low-birth-weight (LBW) infants can promote better behavioral and developmental outcomes. STUDY DESIGN: In this historical control study, 26 infants in the KC group (GA: 34.3+/-2.5 weeks, BW: 1833.9+/-167.6 g) and 27 infants in the comparison group who received the standard medical-nursing care (34.6+/-2.3 weeks, 1850.9+/-156.7 g) were analyzed by the Neonatal Behavioral Assessment Scale (NBAS) at 40 weeks of postmenstrual age, the Bayley Scales of Infant Development and the Carey's Infant Temperament Questionnaire (ITQ) at 6 and 12 months corrected ages. RESULTS: KC infants had significantly higher NBAS scores in Orientation, State Regulation, and Supplementary items; lower Intensity scores and higher Mood scores at 6 months on the ITQ; and higher Bayley Scales score at 12 months. CONCLUSION: KC effectively promoted neonatal behavioral organization and enhanced developmental outcome over the first year of life for LBW infants.

Behavioral and psychophysiologic assessment of the preterm infant.

Cardiorespiratory activity was recorded during attentional responsivity on the Brazelton scale in term and preterm infants. Preterm infants showed less heart rate deceleration, less heart rate variance and less power in the ECG spectrum at frequencies associated with respiratory sinus arrhythmia and oscillations in blood pressure: A lower threshold for attentional stimulation in the preterm infant may trigger a CNS mechanism that protects the infant from stimulus overload. Study of the coordination between behavioral and physiological reactivity provides an understanding of the role of the CNS in mediating information processing.

Specificity of preventative pediatric intervention effects in early infancy.

The efficacy of two contrasting short term preventative interventions administered to a heterogeneous sample of new mothers during the perinatal period was evaluated. The first was infant-centered and used the Brazelton Neonatal Behavioral Assessment Scale (NBAS) as a method of highlighting newborn behavior to new mothers. The second was mother-centered and consisted of an in-depth interview focused on the mothers' concerns about parenting. Mothers were randomized into groups and were administered intervention by experienced clinicians at 3 days in the hospital and again at 14 and 30 days at home. Effects of intervention on maternal reports of parenting stress, mother-infant interactive behavior, and infant developmental quotient were evaluated at 4 months infant age. It was hypothesized that participation in the infant-centered intervention would be related to more positive maternal and infant outcomes at 4 months. It was also expected that the impact of each intervention would be moderated by differences in maternal and infant risk and parity. Hierarchical multiple regressions controlling for risk and parity yielded no significant main effects of intervention at 4 months. However, significant interactions of intervention with parity and risk were observed, indicating that intervention was beneficial for specific groups of mothers. These data suggest that early intervention should be tailored to the needs of individual groups of mothers.

Issues for working parents.

Not only are infants of working parents at risk if we do not provide optimal substitute care for them, their parents will suffer as well. The opportunity to strengthen the family, rather than weaken it, can be provided around a new baby if such issues as parental leave, quality infant care, and attention to key emotional factors in the early separation of parents and infant are addressed.

The Touchpoints Pediatric Asthma Program.

The Brazelton Touchpoints Center at the Child Development Unit, Children's Hospital, Boston, MA, designed a program intended to change the way asthma is managed in medical offices across the United States. This program was recently implemented at five pediatric asthma practices in the Chicago area where asthma prevalence is alarmingly high.