The placenta contributes to activation of the renin angiotensin system in twin-twin transfusion syndrome.

The renin-angiotensin system (RAS) in twin-twin transfusion syndrome (TTTS) is up-regulated in the donor fetus's kidneys, but down-regulated in the recipient's. Ultrasonographic and echocardiographic features suggest that the recipient is also exposed to RAS components. In this study we investigated the role and origin of RAS components in the recipient fetus. Monochorionic diamniotic (MCDA) pregnancies were recruited from a tertiary fetal medicine service. Cord blood was collected from MCDA twins (TTTS and control non-TTTS) at delivery for renin and angiotensin II immunoassays. Placental tissue was flash-frozen for mRNA and protein expression or formalin-fixed for immunohistochemistry. Archival placenta and kidney samples were used for immunohistochemistry and in-situ hybridization. Plasma renin levels were elevated (p<0.05) in recipients (median 201 pg/ml, range 54-315 pg/ml) and donors (125 pg/ml, 25-296) with TTTS compared to controls (2.5 pg/ml, 1.1-1.5 pg/ml). The same was found with angiotensin II with high levels in both recipients (300.5 pg/ml, 86.1-488 pg/ml) and donors (239 pg/ml, 76.6-422) compared to controls (169.5 pg/ml, 89-220 pg/ml, p<0.05). Renin mRNA expression, and protein appeared qualitatively higher in the placental territory of the recipient compared to that of the donor and non-TTTS controls. We conclude that both fetuses in TTTS are exposed to high levels of RAS components; these appear to be produced from different sites, namely the kidney of the donor, and the placenta of the recipient. Given the markedly different phenotypes in the genetically identical fetuses with TTTS, we suggest that the source of RAS components may influence their clinical manifestations.

Inflammatory cytokines in juvenile idiopathic arthritis: effects on physical growth and the insulin-like-growth factor axis.

OBJECTIVE: To investigate the relationship between markers of inflammation with physical growth and systemic markers of GH secretion in JIA. DESIGN: This is a cross sectional prospective study of patients with JIA recruited during therapeutic arthrocentesis of 17 children with JIA (F,10): 8 oligoarticular (OJIA) and 9 polyarticular (PJIA). RESULTS: Median adjusted height (AHt) SDS was -0.3 (-2.2 to 1.6). Serum ALS SDS (median -1.3, range -2.7 to -0.6) was reduced compared with serum IGFBP-3 SDS (median -0.5, range -7.7 to 2.3) and IGF-1 SDS (median -0.2, range -0.5 to 0.5). Log serum IL5 (95% CI -3.25, -0.81) and log serum IL15 (95% CI -9.58, -4.10) were independent factors associated with AHt SDS. Inflammatory cytokines individually showed no association with IGF-1, IGFBP-1, -2, -3 and ALS. CONCLUSION: Children with JIA and mild degree of growth retardation show decreased ALS and IGFBP-3. Cytokines did not show an association to systemic markers of GH secretion. However, this study reports the novel, preliminary association between serum levels of IL5 and IL15 and the extent of short stature.

New bicyclam-AZT conjugates: design, synthesis, anti-HIV evaluation, and their interaction with CXCR-4 coreceptor.

We report the synthesis of mono- and bis-tetraazamacrocycle-AZT conjugates. All new compounds were screened for their ability to inhibit HIV-1 replication in MT4 cell line and were compared to AZT alone. It appears that N-protected covalent prodrugs are equipotent to AZT as inhibitor of HIV replication, while N-deprotected analogues exhibit both higher activity and selectivity against HIV-infected cells. The most active antiviral compounds 27, 28, 34, and 35 were then tested for their binding capability to CXCR-4 receptor. N-Boc analogues 27 and 34 were only weakly effective; in contrast, N-deprotected conjugates 28 and 35 were antagonists to 12G5 mAb binding until 0.05 and 5 microg/mL, respectively. The stability of compound 28 in human plasma was evaluated, and half-life was found to be approximately 8 h in the described conditions. All these results seem to demonstrate the confidence of our prodrug approach, with analogue 28 emerging as the best candidate as lead compound in HIV-1 polytherapy perspective.

A randomised controlled trial of morphine versus phenobarbitone for neonatal abstinence syndrome.

BACKGROUND: The incidence of neonatal abstinence syndrome (NAS) has increased 10-fold over the last decade in Glasgow. In the Princess Royal Maternity Hospital, it now accounts for 17% of special care baby unit (SCBU) admissions. OBJECTIVE: To compare opiate replacement therapy (morphine sulphate) with the present standard treatment (phenobarbitone) for management of NAS. The primary study end point was duration of pharmaceutical treatment. Secondary end points were the requirement for additional drugs and the requirement for SCBU admission. DESIGN: Double blind, randomised controlled clinical trial. METHODS: Differential diagnoses were excluded, and two consecutive Lipsitz scores > 4 defined NAS requiring treatment. Infants were randomised to receive morphine sulphate or phenobarbitone. Treatments were identical in appearance, odour, and volume. Increments, decrements, and discontinuation of treatments were protocol driven. RESULTS: Seventy five infants participated. All mothers received opiate replacement therapy (methadone) during pregnancy and most used other drugs (n = 62, 83%). No significant difference in maternal drug use patterns was observed between treatment groups. Median treatment duration was four days shorter with opiate replacement (8 v 12 days, Mann-Whitney U test, p = 0.02). Phenobarbitone treated infants tended to require second line treatment (47% v 35%, chi(2) test, p = 0.11) and SCBU admission (62% v 30%, chi(2) test, p = 0.04) more often. CONCLUSIONS: Opiate replacement therapy appears to be superior for management of symptomatic NAS when maternal opiate use is prevalent. The shorter treatment duration and lower requirement for higher intensity nursing may have significant cost implications. Tailoring NAS treatment to local maternal drug use may result in similar benefits.

Familial aplasia cutis congenita.

Two siblings with extensive aplasia cutis congenita of the extremities are described. These lesions healed spontaneously and in the older sibling at four years follow-up there was minimal scar tissue at the affected sites.

Neonatal listeriosis in Scotland.

Thirty-one cases of neonatal listeriosis occurred in Scotland between 1 January 1987 and 31 August 1989. In each of these, Listeria monocytogenes was cultured from blood, cerebrospinal fluid or both. No case of neonatal listeriosis occurred between 1 September 1989 and 31 July 1990, a statistically significant difference in incidence (p less than 0.001). A possible explanation may be that pregnant women adjusted their diet in accordance with the advice given by the Chief Medical Officer in his letter to General Practitioners of February 1989 entitled 'Food and Listeriosis'. Full clinical features and outcome of nineteen of the thirty-one cases were obtained from the paediatricians responsible for their care. Of these, sixteen were early onset and three late onset cases. There were eight deaths, all early onset cases. Two severely asphyxiated infants could not be resuscitated and of the five infants who required assisted ventilation from birth, four died. Four early onset and three late onset cases presented with pyogenic meningitis and all seven recovered without sequelae. It is concluded that when neonatal listeriosis presents as an asphyxiated infant who, after resuscitation, requires assisted ventilation, a fatal outcome is probable. When neonatal listeriosis presents as pyogenic meningitis, recovery without sequelae is the usual outcome.

Toxic shock syndrome (TSS) in children.

Toxic shock syndrome is uncommon in the prepubertal age group. Two children presented with pyrexia, macular erythroderma, vomiting, hypotension and rapid deterioration of consciousness. One child had severe neurological involvement. The diagnosis of toxic shock syndrome was established in both cases by the exclusion of other causes and by culturing staphylococcus aureus. We postulate that the neurological manifestations were caused by a direct neurotoxic action of the staphylococcal-produced toxin. Both children made a complete recovery.

Tuberculous meningitis in children: a review of 15 cases.

Tuberculosis meningitis (TBM) is the most serious form of infection with Mycobacterium tuberculosis. Between 1968 and 1986 15 children (five boys and 10 girls) were seen at the Royal Hospital for Sick Children, Glasgow, because of TBM. Fourteen children were Caucasian and one was Asian. The mean age at presentation was two years. None had been given BCG vaccination. In 12 children close contact with other cases of tuberculosis was reported. The signs and symptoms which helped in the diagnosis are discussed together with the initial CSF findings, results of mantoux testing and chest X-rays. Three children had unusual modes of presentation. All children were treated with chemotherapy though the drug combinations, route of administration and therapy varied from case to case. Steroids were used in nine children. Five children required neurosurgical intervention. Two children died and of the survivors six had serious sequelae. Five children made a complete recovery. The outcome of TBM depended on the duration of symptoms prior to the onset of therapy, on the neurological status reached at the time of diagnosis and the age of the child. The roles of chemotherapy, steroids and neurosurgery in the management of TBM are discussed. The need for routine BCG vaccination of all neonates is examined.

Ocular complications in juvenile chronic arthritis (JCA).

Fifty-six children with JCA have attended the Rheumatology Clinic at the Royal Hospital for Sick Children, Glasgow, over the past five years. Eleven have monoarthritis, 21 pauciarthritis and 24 polyarthritis. Seven children with pauciarthritis and one with monoarthritis developed ocular complications. Of these six were girls. In six children the arthritis preceded the uveitis. In one child arthritis and uveitis presented at the same time and in another the uveitis preceded the arthritis by one year. All were treated with steroids (7 topically, 1 systemically) and topical mydriatic agents. After an initial response the uveitis persisted as a low-grade inflammation gradually leading to secondary complications and increasing loss of vision. Only two patients enjoy normal vision at present. The importance of routine slit-lamp microscopy in all children with JCA is stressed, especially in those with pauciarthritis and antinuclear antibodies.

[Interleukin 4 increases lymphocytic adhesion to allogenic endothelium through a dependent protein kinase A]. / L'interleukine 4 augmente l'adhésion lymphocytaire à l'endothélium allogénique par une voie protéine kinase A dépendante.

Interleukin 4 and tumor necrosis factor alpha increase endothelial cell adhesiveness for lymphocytes by promoting adhesion molecules synthesis. We have shown that intracellular mechanisms, which transduce IL 4--but not TNF alpha--signal, involve a protein kinase A.

[Adhesion of lymphocytes to allogenic endothelium. Four different pathways]. / Adhésion des lymphocytes à l'endothélium allogénique. Quatre voies différentes.

Lymphocyte adhesion to allogeneic endothelium is a critical step in graft rejection. To further characterize the expression and the regulation of adhesion molecules involved in this process, we stimulated endothelial cells with Interleukin 4 or tumor necrosis factor alpha (TNF alpha) and blocked lymphocyte adhesion to stimulated-endothelial cells with monoclonal antibodies. We demonstrated that lymphocytes bound endothelial cells by at least four adhesion pathways: a) a LFA 1/ICAM 1 dependent pathway on TNF alpha-stimulated endothelial cells, b) a LFA 1 dependent/ICAM 1 independent pathway on unstimulated or IL 4-stimulated endothelial cells, c) a VLA 4/VCAM 1 dependent pathway on unstimulated and stimulated-endothelial cells, d) a CD 2 dependent/LFA 3 independent pathway on IL 4 and TNF alpha-stimulated endothelial cells.

Development of monoclonal antibodies to pre-haptoglobin 2 and their use in an enzyme-linked immunosorbent assay (ELISA).

Haptoglobins (HPs) are alpha 2-globulin proteins that bind free hemoglobin in plasma to prevent oxidative damage. HPs are produced as preproteins that are proteolytically cleaved in the ER into alpha and beta chains prior to forming mature, functional tetramers. Two alleles exist in humans (HP1 and HP2), therefore three genotypes are present in the population, i.e., HP1-1, HP2-1, and HP2-2. A biochemical role for nascent haptoglobin 2 (pre-haptoglobin 2 or pre-HP2) as the only known modulator of intestinal permeability has been established. In addition, elevated levels of serum pre-HP2 have been detected in multiple conditions including celiac disease and type I diabetes, which are believed to result in part through dysregulation of the intestinal barrier. In this study, we report the development of a monoclonal antibody that is specific for pre-HP2 with a binding affinity in the nanomolar range. Additional antibodies with specificities for preHP but not mature haptoglobin were also characterized. A sandwich enzyme-linked immunosorbent assay (ELISA) was established and validated. The ELISA showed high specificity for pre-HP2 even in the presence of excess pre-HP1 or mature haptoglobins, and has excellent linearity and inter- and intra-assay reproducibility with a working range from 3.1ng/mL to 200ng/mL. Testing of sera from 76 healthy patients revealed a non-Gaussian distribution of pre-HP2 levels with a mean concentration of 221.2ng/mL (95% CI: 106.5-335.9ng/mL) and a median value of 23.9ng/mL. Compared to current approaches, this ELISA offers a validated, monoclonal-based method with high sensitivity and specificity for measuring pre-HP2 in human serum.

Generation and molecular characterization of a monoclonal antibody reactive with conserved epitope in sphingomyelinases D from Loxosceles spider venoms.

We report the production of a neutralizing monoclonal antibody able to recognize the venoms of three major medically important species of Loxosceles spiders in Brazil. The mAb was produced by immunization of mice with a toxic recombinant L. intermedia sphingomyelinase D {SMases D isoform (rLiD1)} [1] and screened by enzyme-linked immunosorbent assay (ELISA) using L. intermedia, L. laeta and L. gaucho venoms as antigens. One clone (LiD1mAb16) out of seventeen anti-rLiD1 hybridomas was cross-reactive with the three whole Loxosceles venoms. 2D Western blot analysis indicated that LiD1mAb16 was capable of interacting with 34 proteins of 29-36kDa in L. intermedia, 33 in L. gaucho and 27 in L. laeta venoms. The results of immunoassays with cellulose-bound peptides revealed that the LiD1mAb16 recognizes a highly conserved linear epitope localized in the catalytic region of SMases D toxins. The selected mAb displayed in vivo protective activity in rabbits after challenge with rLiD1. These results show the potential usefulness of monoclonal antibodies for future therapeutic approaches and also opens up the perspective of utilization of these antibodies for immunodiagnostic assays in loxoscelism.

Cerebrospinal Aß11-x and 17-x levels as indicators of mild cognitive impairment and patients' stratification in Alzheimer's disease.

In the present work, the concentrations of Aß11-x and Aß17-x peptides (x=40 or 42), which result from the combined cleavages of ß-amyloid precursor protein (AßPP) by ß'/α or α/γ-secretases, respectively, were assessed in cerebrospinal fluid (CSF) samples from patients with Alzheimer's disease (AD) or mild cognitive impairment (MCI). Specific multiplexed assays were set up using new anti-40 and anti-42 monoclonal antibodies (mAbs) for the capture of these N-truncated Aß peptides and anti-11 or anti-17 mAbs for their detection. The specificity, sensitivity and reproducibility of such assays were assessed using synthetic peptides and human cell models. Aß11-x and Aß17-x were then measured in CSF samples from patients with AD (n=23), MCI (n=23) and controls with normal cognition (n=21). Aß11-x levels were significantly lower in patients with MCI than in controls. Compared with the combined quantification of Aß1-42, total Tau (T-Tau) and phosphorylated Tau (P-Tau; AlzBio3, Innogenetics), the association of Aß11-40, Aß17-40 and T-Tau improved the discrimination between MCI and controls. Furthermore, when patients with MCI were classified into two subgroups (MCI ≤1.5 or ≥2 based on their CDR-SB (Cognitive Dementia Rating-Sum of Boxes) score), the CSF Aß17-40/Aß11-40 ratio was significantly higher in patients with CDR-SB ≤1.5 than in controls, whereas neither Aß1-42, T-Tau nor P-Tau allowed the detection of this subpopulation. These results need to be confirmed in a larger clinical prospective cohort.