BOS is associated with decreased HDAC2 from steroid resistant lymphocytes in the small airways.

Immunosuppression therapies including corticosteroids fail to prevent bronchiolitis obliterans syndrome (BOS), primarily a disease of the small airways, following lung transplantation. We reported increases in steroid-resistant proinflammatory lymphocytes and their loss of histone deacetylase 2 (HDAC2), an important mediator of steroid action, in the blood of stable lung transplant recipients. We noted similar increases in the steroid-resistant lymphocytes in both the blood and small airways in BOS compared with the large airways. We hypothesized that these small airway cells would also exhibit a loss of HDAC2, and that these changes could be reversed by treatment with theophylline (HDAC2 activator). Blood, bronchoalveolar lavage and large and small airway brushings were collected from lung transplant patients with BOS (n = 12) or stable lung function (n = 18) and healthy aged-matched controls (n = 13). Intracellular proinflammatory cytokines [interferon (IFN-γ) and tumour necrosis factor (TNF)-α and HDAC2 were measured in CD8+ T, natural killer (NK) T-like and NK cells from cultured small airway brushings ± 5 mg/l theophylline ± 1 µM prednisolone using flow cytometry. Increased small airway CD8 T, NK T-like and NK cells were identified in BOS versus stable transplant and controls. In BOS, these cells exhibited increased IFN-γ/TNF-α and a loss of HDAC2. HDAC2 expression by small airway CD8+ T cells correlated with forced expiratory volume in 1 s (FEV1 ) (R = 0·880, P = 0·031). Theophylline and prednisolone synergistically up-regulated HDAC2 in CD8+ T cells. BOS is associated with loss of HDAC2 from steroid-resistant proinflammatory CD8+ T, NK T-like and NK cells in the small airways. Therapeutically increasing HDAC2 in these lymphocytes may reduce steroid resistance and improve graft survival.

The impact of immunoglobulin in acute HIV infection on the HIV reservoir: a randomized controlled trial.

OBJECTIVES: Antiretroviral therapy (ART) during acute HIV infection (AHI) restricts the HIV reservoir, but additional interventions are necessary to induce a cure. Intravenous immunoglobulin (IVIG) is not HIV-specific but is safe and temporarily reduces the HIV reservoir in chronic HIV infection. We present a randomized controlled trial to investigate whether IVIG plus ART in AHI reduces the HIV reservoir and immune activation compared with ART alone. METHODS: Ten men with AHI (Fiebig II-IV) initiated ART (tenofovir, entricitabine, ritonavir boosted darunavir and raltegravir) at HIV-1 diagnosis and were randomized to ART alone or ART plus 5 days of IVIG, once virally suppressed (week 19). Blood samples were evaluated for viral reservoir, immune activation, immune exhaustion and microbial translocation. Flexible sigmoidoscopy was performed at weeks 19, 24 and 48, and gut proviral DNA and cell numbers determined. RESULTS: IVIG was well tolerated and no viral blips (> 50 HIV-1 RNA copies/mL) occurred during IVIG therapy. From baseline to week 48, total HIV DNA in peripheral blood mononuclear cells (PBMCs) (cases: -3.7 log10 copies/106 CD4 cells; controls: -3.87 log10 copies/106 CD4 cells) declined with no differences observed between the groups (P = 0.49). Declines were observed in both groups from week 19 to week 48 in total HIV DNA in PBMCs (P = 0.38), serum low copy RNA (P = 0.57) and gut total HIV DNA (P = 0.55), but again there were no significant differences between arms. Biomarkers of immune activation, immune exhaustion and microbial translocation and the CD4:CD8 ratio were similar between arms for all comparisons. CONCLUSIONS: Although safe, IVIG in AHI did not impact total HIV DNA, immune function or microbial translocation in peripheral blood or gut tissue.

Erythropoietin-induced neuroprotection requires cystine glutamate exchanger activity.

Erythropoietin (Epo) has been used for many years in neonates for the treatment of anemia of prematurity. Epo has also been proposed for treatment of neonatal brain injury, as mounting evidence suggests neuroprotective properties for Epo. However, Epo's neuroprotective mechanism of action is poorly understood. In this study we hypothesized that Epo may confer neuroprotection by enhancing cellular redox defense brought about by cellular glutathione (GSH). This was examined in cultures of differentiated cortical neural stem cells and using the B104 cell line as model systems. Our data shows that Epo causes a time- and dose-dependent increase in expression and activity of system Xc(-), the transporter responsible for uptake of cystine for the production of glutathione. Cystine uptake increases 3-5 fold in differentiated neural stem cells and B104 cells treated with Epo. Exposure of cells to 100 microM kainate suppressed cellular GSH and caused excitotoxicity, but GSH levels and cell viability were completely restored by Epo in the continued presence of kainate. This rescue effect of Epo vanished if system Xc(-) was inhibited pharmacologically using S4-CPG in the presence of Epo leading to marked cell death of B104 cells and cultured mouse cortical neural stem cells. This could also be achieved using xCT siRNA to decrease xCT expression. This data suggests that system Xc(-) activity and protein expression are positively regulated by Epo directly explaining its neuroprotective effect.

Naloxone-precipitated withdrawal jumping in 11 inbred mouse strains: evidence for common genetic mechanisms in acute and chronic morphine physical dependence.

Physical dependence is a widely known consequence of morphine intake. Although commonly associated with prolonged or repeated morphine administration, withdrawal symptoms can be elicited even after a single prior morphine exposure. What remains contentious is the extent to which physical dependence following acute and chronic morphine treatment is mediated by common physiological substrates and, accordingly, represent distinct syndromes. The genetic relationship between acute and chronic morphine dependence was thus presently studied by comparing mice of 11 inbred strains (129P3, A, AKR, BALB/c, C3H/He, C57BL/6, CBA, DBA/2, LP, SJL, and SWR) for naloxone-precipitated withdrawal jumping responses using three subcutaneous morphine administration paradigms: acute (single injection) or chronic (three daily morphine injections for 4 days) injection, or chronic infusion (7 days via implanted osmotic minipumps). Although there were differences in the magnitude of withdrawal jumping between the three different morphine administration paradigms, large and significant strain differences were observed for each. In addition, the same strains were unusually sensitive or, conversely, altogether refractory to withdrawal jumping across all morphine treatment conditions. Overall, strain jumping means between acute and chronic dependence paradigms displayed a high degree of genetic correlation (r=0.87-0.95). The significant correlation between chronic morphine injection and continuous morphine infusion discounts the possible confounding effect of contextual learning and spontaneous withdrawal between chronic injections on the assessment of naloxone-precipitated withdrawal. Substantial heritability was also observed for acute and both paradigms of chronic dependence, with estimates ranging from h(2)=0.53 to 0.70. The present demonstration of a strong genetic correlation between physical dependence to morphine following acute and chronic treatment implies that genes associated with variable sensitivity in the two traits are the same, and is suggestive of shared physiological substrates. The data also demonstrate that the differential genetic liability to morphine physical dependence begins with, and is predicted by, the first morphine exposure.

Assessment of acute and chronic morphine dependence in male and female mice.

The present study compared male and female mice for frequency of naloxone-precipitated jumping and naloxone ED(50) values, two common indices of physical dependence, following acute and chronic morphine administration. Both sexes displayed a positive dose-response relationship between acute morphine and naloxone doses and jumping frequency. There was a significant main effect of sex, with mean jumping frequencies greater in males. The naloxone ED(50) estimate was also fourfold lower in males, indicating greater withdrawal sensitivity than females. Jumping frequencies were similar in male and female saline-treated control mice, discounting initial sex differences as a significant factor in the unequal magnitude and sensitivity in acute morphine dependence between sexes. In contrast, males and females displayed similar mean withdrawal jumping frequencies and naloxone ED(50) values after 3 days of morphine injections. Sex difference in withdrawal jumping was also not observed when morphine treatment was increased to 7 days via daily injection or continuous subcutaneous infusion. The present study demonstrates the development of greater physical dependence in male relative to female mice following acute but not chronic morphine administration.

Morphine tolerance and dependence in nociceptin/orphanin FQ transgenic knock-out mice.

It has been hypothesized that morphine tolerance and dependence in mice following chronic exposure may reflect increased compensatory activity of antiopioid systems. The endogenous peptide nociceptin/orphanin FQ has been shown to have anti-opioid effects, for example antagonizing morphine analgesia. Moreover, chronic morphine administration increases synthesis of the peptide, and morphine tolerance and dependence can be attenuated or reversed by antagonists and agonists of the nociceptin/orphanin FQ receptor, respectively. The present study seeks to confirm a role for nociceptin/orphanin FQ in opioid tolerance and dependence by comparing morphine ED(50) values and naloxone-precipitated withdrawal jumping in mice homozygous (knock-out) and heterozygous for a null mutation of the Npnc1 gene encoding the nociceptin/orphanin FQ propeptide, and their wild type littermates, following chronic morphine exposure. Relative to morphine-naive control mice, significant rightward shifts in the morphine dose-response curve, resulting in increased morphine ED(50) values (approximately two to three-fold), was observed for all genotypes following three days of repeated systemic morphine injections. However, no differences between genotypes in the magnitude of tolerance were observed. In contrast, knock-out mice displayed significantly increased naloxone-precipitated withdrawal jumping relative to heterozygous and wild-type mice following implantation with a morphine pellet (25mg) for 72h. Use of nociception/orphaninFQ transgenic knock-out mice thus demonstrate the differential involvement of nociceptin/orphanin FQ in morphine tolerance and dependence.

Selecting patients for combined femorofemoral bypass grafting and iliac balloon angioplasty and stenting for bilateral iliac disease.

PURPOSE: This study examines the selection of patients for combined femorofemoral bypass (FFB) grafting and iliac balloon angioplasty (IBA) and stenting for bilateral iliac occlusive disease (successively or simultaneously) and the correlation of the length and location of stenoses of the donor iliac artery to the success of FFB grafts. METHODS: Forty-one patients with long iliac occlusion and significant contralateral iliac stenosis were treated with combined FFB grafting and IBA and stenting, which were performed simultaneously or percutaneously within 1 to 2 days before surgery. Stenting was performed for suboptimal IBAs. IBA/graft patency was evaluated by duplex scanning/ankle-brachial index at 1, 3, 6, and 12 months and every 12 months thereafter. A life-table analysis of patency was performed, according to the length of stenosis as classified by the Society of Cardiovascular Interventional Radiology (group A, < 3 cm and 3-5 cm; group B, > 5 cm). RESULTS: Indications for surgery were limb salvage (22%), rest pain (44%), and claudication (34%). The mean follow-up time was 34.1 months. Perioperative complications were 7% for group A versus 62% for group B (P = .0007) with no perioperative deaths or amputations. Stenting was needed in 12 of 13 patients (92%) in group B versus four of 28 patients (14%) in group A (P < .0001) and in 11 of 12 external iliac artery lesions versus five of 29 common iliac artery lesions (P < .0001). The overall early success rate was 100% for group A and 62% for group B (P = .0028). The primary patency rates at 1, 2, and 3 years were 96%, 85%, and 85% for group A, respectively, and for group B were 46%, 46%, and 31%, respectively (P < .01). The secondary patency rates for group A at 1, 2, and 3 years were 100%, 96%, and 87%, respectively; and for group B were 62%, 54%, and 27%, respectively (P < .001). The overall primary and secondary patency rates for common iliac and external iliac artery lesions were similar (72% and 72% versus 67% and 75%, respectively). The overall limb salvage rates were 96% for group A and 85% for group B. Seven of 13 patients (54%) of group B, in contrast with 0 of 28 patients in group A, had to undergo a revision of the procedure within 30 days (P < .01). CONCLUSION: Combined use of IBA and stenting and FFB grafting is effective and durable and can be performed simultaneously, if the donor iliac stenosis length is 5 cm or less. Percutaneous transluminal angioplasty/stenting of stenoses of 5 cm or more fail to support FFB grafting in most patients; therefore, their combination should be questioned.

Morphine tolerance after chronic intracerebroventricular injection in male and female mice.

We compared morphine analgesia ED(50) values of male and female mice prior to (Day 1) and after (Day 4) 3 days of intracerebroventricular morphine injections. Increases in ED(50) values from Day 1 to Day 4, indicating tolerance, were of similar magnitude in both sexes. The data suggest that spinal opioid analgesic mechanisms, acting alone or in synergy with supraspinal loci, may contribute to pharmacodynamic explanations of sex differences in systemic morphine tolerance.

A comparison of morphine analgesic tolerance in male and female mice.

Studies comparing morphine tolerance in males and females are rare, and all studies to date have utilized the rat. To generalize from findings with rats morphine tolerance was investigated in male and female mice using the tail-withdrawal test. Three and 7 days of systemic morphine injections produced significant but unequal rightward shifts in the morphine dose-response curve such that females displayed greater increases in analgesic ED(50) values when compared to males. In a separate experiment, males and females displayed similar reductions in morphine analgesic sensitivity when %MPE (maximum possible effect) and %total (area under the curve) were compared after 3 days of morphine. Differences in initial morphine sensitivity between sexes were not observed in either study. The data demonstrate that, in contrast to rats, female mice undergo greater reductions in morphine analgesia relative to males following chronic morphine, but this sex difference may depend on the method of assessing analgesia. Furthermore, the duration and/or cumulative dose of morphine treatment does not affect the expression of sex differences in morphine tolerance.

Sex differences in thermoregulation after acute and chronic morphine administration in mice.

Sex differences in thermoregulation have been reported following acute morphine administration in rats only. This study assessed whether male and female mice also differ in thermoregulatory responses following acute and chronic morphine administration. Females displayed significantly higher baseline colorectal temperature and greater morphine (24mg/kg, s.c.) hypothermia (2. 5-fold) on day 1. Two additional days of morphine treatment did not alter baseline temperature readings on Day 4 in either sex, but significantly reduced the morphine hypothermia relative to Day 1 in a sex-dependent manner. Whereas the morphine hypothermia was completely abolished in males, significant hypothermia was still observed in females. Acute and chronic saline injections had no effect on colorectal temperature. The data demonstrate sex differences in the thermoregulatory responses to acute and chronic morphine administration in mice.

Calcium kinetics in children with osteogenesis imperfecta type III and IV: pre- and post-growth hormone therapy.

Children with osteogenesis imperfecta (OI) type III and type IV were studied using a (42)Ca stable isotope technique. Serum dilution kinetics of (42)Ca were studied pre- and post-growth hormone (GH) treatment in 9 OI III (age range 5-9 years) and 8 OI IV patients (age range 5-12 years). Each subject was studied twice: at baseline and following GH therapy (range 1-1.5 years). Isotopic enrichments of (42)Ca were followed over 7 days using thermal ionization mass spectrometry. A binding site model, which describes reversible and irreversible binding of calcium (Ca) ions to postulated short- and long-term binding sites in bone, was used to analyze the kinetic data. In type III patients, GH treatment (1) increased the fraction of short-term binding sites, theta (0.777 +/- 0.112 versus 0.877 +/- 0.05, respectively; P = 0.034); (2) increased the apparent half-life of a Ca ion attached to the long-term binding site by 76% (P = 0. 009); (3) although not statistically significant (P = 0.098), a trend toward an increased growth rate was observed with increasing change in theta (Deltatheta); (4) patients experienced a 75% increase in growth rate during the first 6 months of treatment. In type IV patients, GH treatment increased the apparent half-life of a Ca ion attached to the long-term binding site by 83% (P = 0.048), however, no trend toward an increased growth rate was observed with increasing Deltatheta in these patients. These significant changes in Ca binding to bone may influence growth in type III patients.

Effect of growth hormone treatment on calcium kinetics in patients with osteogenesis imperfecta type III and IV.

Using a dual stable isotope technique, the effect of growth hormone (GH) on whole body calcium (Ca) metabolism was studied in children (ages 5-14 years) with type III (n = 9) and IV (n = 8) osteogenesis imperfecta. Each subject was studied twice: at baseline and following a GH (0.1-0.2 U/kg per day) treatment period of 1-1.5 years. Subjects were given 42Ca intravenously and 44Ca orally. The sera and urine 42Ca and 44Ca isotopic enrichments were followed over 7 days using thermal ionization mass spectrometry. The SAAM program was used to fit a three-compartment model to the tracer data. No significant differences were observed between: (1) children with type III and IV disease; or (2) baseline studies of boys and girls within each disease type. However, GH treatment significantly increased: (1) the exchangeable calcium pool (EP) in type III patients (2086 vs. 4422 mg/day, p = 0.02); and (2) the parameter associated with bone calcium accretion in type IV patients (Vo+: 973 vs. 1560 mg/day,p = 0.03) with boys responding with a significantly greater increase than girls (p = 0.008). Although not statistically significant, a trend toward an increase in Vo+ in type III patients and in EP in type IV was observed following treatment. Our observations imply that more Ca was available for bone mineralization following GH treatment in these subjects.

A single surgeon's experience with the Delaire palatoplasty.

The purpose of this review was to evaluate the clinical outcomes regarding velopharyngeal insufficiency and fistulization in patients with cleft palate who underwent primary repair with the one-stage Delaire palatoplasty. All patients who had a primary Delaire-type palatoplasty performed by the senior surgeon over a 10-year period (1988 to 1998) were studied. During this period, each consecutive patient with an open palatal cleft underwent the same type of repair by the same surgeon. Speech quality and velopharyngeal competence as determined by a single speech pathologist were recorded. A total of 95 patients were included in this series. The average length of follow-up was 31 months (range, 1 to 118 months). Average age at time of surgery was 13.3 months (range, 6 to 180 months). Thirty-one patients (32.6 percent) had significant associated anomalies. The average length of hospital stay was 1.9 days (range, 1 to 8 days) with a trend in recent years toward discharge on postoperative day 1. There were no intraoperative complications, either surgical or anesthetic. Three patients (3.2 percent) developed palatal fistula; none of them required repair. Six patients (6.3 percent) had velopharyngeal incompetence. In patients with more than 1 year of follow-up, the incidence of velopharyngeal incompetence was 9.2 percent (6 of 65). The incidence of fistula after the Delaire palatoplasty was lower than usually reported. The incidence of velopharyngeal incompetence requiring pharyngoplasty was equal to or lower than that seen after other types of palatoplasty, suggesting superior soft-palate muscle function attributable to approximation of the musculus uvulae. The Delaire palatoplasty results in a functional palate with low risk for fistula formation and velopharyngeal incompetence.

Noise-induced neurologic disturbances in divers exposed to intense water-borne sound: two case reports.

Divers may be exposed to intense noise underwater. Two cases of neurologic disturbances during experimental exposures to 15 min of continuous underwater sound are described. Sound exposure in the first case consisted of a warble tone with center frequency of 240 Hz and a sound pressure level of 160 dB re 1 microPa. Symptoms during exposure consisted of somnolence, lightheadedness, and an inability to concentrate. No apparent effect on hearing was noted. In the second case, a center frequency of 1,000 Hz at 181 dB was used. Lightheadedness, inability to concentrate, agitation, and head vibrations were noted during the exposure. The diver also exhibited a temporary auditory threshold shift of 19.2 dB. In both cases, overt symptoms resolved within 30 min after exposure, but both divers reported recurrent symptoms days to weeks after the exposures. Medical histories and examinations, assessment of dive profiles, and breathing gas analysis failed to support a source other than the sound exposures to account for the symptoms observed. Potential mechanisms for the described symptoms are discussed.

Alterations in swim stress-induced analgesia and hypothermia following serotonergic or NMDA antagonists in the rostral ventromedial medulla of rats.

Serotonergic, NMDA, or opioid antagonists in the rostral ventromedial medulla (RVM) reduce morphine analgesia elicited from the periaqueductal gray (PAG). Continuous (CCWS) and intermittent (ICWS) cold-water swims elicit respective naltrexone-insensitive and naltrexone-sensitive analgesic responses. CCWS analgesia is reduced by systemic NMDA receptor antagonism and by systemic, but not intrathecal serotonergic antagonism. ICWS analgesia is reduced by both systemic and intrathecal serotonergic antagonism, but unaffected by systemic NMDA antagonism. The present study evaluated whether serotonergic (methysergide: 5-10 microg) or competitive [AP7 (2-amino-7-phosphonoheptanoic acid): 0.01-0.1 microg] or non-competitive [MK-801 (dizocilipine maleate): 0.3-3 microg] NMDA antagonists in the RVM altered CCWS and ICWS analgesia and hypothermia as well as basal nociceptive latencies. Methysergide in the RVM significantly potentiated CCWS, but not ICWS analgesia. In contrast, AP7 in the RVM significantly potentiated ICWS analgesia. Antagonist-induced changes in either hypothermia or basal nociception failed to account for any alterations in stress-induced analgesia. These data suggest that serotonergic, but not NMDA, receptors in the RVM may mediate collateral inhibition between mesencephalic morphine analgesia and naltrexone-insensitive CCWS analgesia.

Cancer support nurses: a co-ordinating role in cancer care.

Cancer support nurses (CSNs) have a key co-ordinating role in the cancer services in North Lancashire and South Lakeland. The working practices of the three CSNs in North Lancashire are described in detail. There was a total cumulative caseload of 604 patients in September 1996, with 397 referrals in the previous 12-month period. There were 267 deaths during this time. CSNs are an essential element of cancer service provision. A ratio of one CSN to 40,000 population gives a manageable workload, provided there is an infrastructure of supportive care to which patients, carers and bereaved can be referred.

Panic with a twist: an unusual presentation of combined psychiatric and neurologic symptoms in a tactical jet aviator.

A 28-yr-old Naval F-14 aviator presented with complaints of flight-related anxiety occurring intermittently over an 18-mo period. Symptoms included sensation of strangeness, concern over the welfare of his radar intercept officer, flushing, nausea, and intense need to immediately land the aircraft. He also described a 6-mo history of episodes wherein he would see "shooting stars" in the periphery of his vision, accompanied by dizziness and disorientation. These latter attacks were always precipitated by head turning, usually in combination with positive Gz maneuvers, and were relieved by head straightening. The anxiety symptoms were consistent with a form of panic attack, but the neurological symptoms provoked further workup. Magnetic resonance cerebral angiogram demonstrated a dominant right vertebral artery and hypoplastic left vertebral artery. All symptoms resolved once the aviator was removed from flying the aircraft. After a year of follow-up with an aviation psychiatrist, he remained asymptomatic and was reassigned to maritime patrol aircraft. This case illustrates a difficult diagnostic, therapeutic, and disposition challenge. This aviator suffered from a complex interaction of neurologic and psychiatric manifestations having a common inciting stimulus, namely flying the F-14 Tomcat. A promising aviation career was preserved upon removal of that stimulus.

Role of the flight surgeon in fighter squadron Arctic deployments.

Operational fighter squadrons frequently find themselves deployed to semi-isolated stations in the Arctic. This paper discusses the major issues necessary for flight surgeon consideration. In particular, the areas of pre-deployment planning, preparing for the worst, routine operations, and post deployment actions are discussed. A recent month-long deployment of a 12-ship squadron of F18s with support elements from Bagotvi le, Canada, to Evenes, Norway, is examined. A proposed kit list to support a similar deployment to a semi-isolated station is provided.

An aviator with an unusual gait: a rare disease teaches some everyday lessons.

Muscular diseases including the dystrophies and myopathies are often incompatible with a variety of occupations including aviation and military duty. Many of these diseases present early in life, are readily diagnosable, and are therefore rare in the aviation community because of pre-screening and selection. Some forms, however, may not present until adulthood during an established aviation career. Furthermore, although initial presentations may be subtle and insidious, the potential occupational and aeromedical ramifications of these diseases can be profound. The following report describes the case of a subjectively asymptomatic career military aviation officer who presented with an unusual gait, and was subsequently determined to have one of the late-presenting muscle disease variants: Anterior compartment Distal Myopathy. The patient's presentation and progression, diagnostic evaluation, prognosis, aeromedical risk and disposition, and issues of occupational and aeromedical significance are discussed.