Enumeration of Gut-Homing β7-Positive,Pathogen-Specific Antibody-Secreting Cells in Whole Blood from Enterotoxigenic Escherichia coli- and Vibrio cholerae-Infected Patients,Determined Using an Enzyme-Linked Immunosorbent Spot Assay Technique

Vibrio cholerae and enterotoxigenic Escherichia coli (ETEC) are noninvasive mucosal pathogens that cause acute watery diarrhea in people in developing countries. Direct assessment of the mucosal immune responses to these pathogens is problematic. Surrogate markers of local mucosal responses in blood are increasingly being studied to determine the mucosal immune responses after infection. However, the volume of blood available in children and infants has limited this approach. We assessed whether an approach that first isolates β7-positive cells from a small volume of blood would allow measurement of the antigen-specific immune responses in patients with cholera and ETEC infection. β7 is a cell surface marker associated with mucosal homing. We isolated β7-expressing cells from blood on days 2, 7, and 30 and used an enzyme-linked immunosorbent spot (ELISPOT) assay to assess the gut-homing antibody-secreting cells (ASCs) specific to pathogen antigens. Patients with ETEC diarrhea showed a significant increase in toxin-specific gut-homing ASCs at day 7 compared to the levels at days 2 and 30 after onset of illness and to the levels in healthy controls. Similar elevations of responses to the ETEC colonization factors (CFs) CS6 and CFA/I were observed in patients infected with CS6- and CFA/I-positive ETEC strains. Antigen-specific gut-homing ASCs to the B subunit of cholera toxin and cholera-specific lipopolysaccharides (LPS) were also observed on day 7 after the onset of cholera using this approach. This study demonstrates that a simple ELISPOT assay can be used to study the mucosal immunity to specific antigens using a cell-sorting protocol to isolate mucosal homing cells, facilitating measurement of mucosal responses in children following infection or vaccination.     

DNA microarray technology for neonatal screening

Modern molecular biology, owing much to the Human Genome Initiative, has elucidated many of the genetic mechanisms underlying heritable metabolic disease. While the use of molecular methods has flourished in research laboratories, complexity and cost have limited their utility in newborn screening. Newborn blood cards provide high quality DNA samples able to provide reliable support to highly multiplexed polymerase chain reactions (PCR). New manufacturing processes have reduced the cost of DNA microarray technology to the point where it is a practical tool for population screening. In a single assay, a DNA microarray facilitates the co-detection of amplification products diagnostic for several genetic diseases. High throughput is achieved with automation at every step, from DNA extraction to detection of hybrids. We suggest that it is both feasible and practical to develop a first-tier newborn screening protocol based upon multiplex PCR and analysis of amplification products using DNA microarrays. Initial data utilizing the model systems of sickle cell disease, α-1-antitrypsin deficiency and Factor V Leiden will be reported.     

Parents'fear regarding fever and febrile seizures

In order to improve the effectiveness of information, we studied parents' perceptions and knowledge about fever and febrile seizures. A questionnaire study was carried out among the parents whose children (n = 230) participated in a randomized controlled trial of ibuprofen to prevent recurrent febrile seizures. Of the 230 parents, 181 (79%) responded to the questionnaire. Of all parents, 45% were afraid or very afraid of fever, which was strongly associated with being afraid of recurrent febrile seizures. Parents of children with a non-West European background were more afraid. The consequences of parental fear included frequent temperature measurements (25% measured five times per day or more), sleeping in the same room (24%) and 13% remained awake at night. Witnessing a febrile seizure is a frightening experience for parents; a majority thought that febrile seizures were harmful, because they look dangerous. Forty-seven percent thought that their child was dying during the initial febrile seizure. On the other hand, reassuring information may be helpful: 21% mentioned it as their reason to consider febrile seizures not harmful. We conclude that parental fear of fever and febrile seizures is a major problem with several negative consequences for daily family life. Adequate provision of information may reduce parental fear. We suggest that information about fever and febrile seizures should be provided to all parents, preferably during their contact with the providers of preventive healthcare. The parents of children with a non-West European origin need extra attention.     

Ontogenetic expression of trk neurotrophin receptors in the chick auditory system.

Neurotrophins and their cognate receptors are critical to normal nervous system development. Trk receptors are high-affinity receptors for nerve-growth factor (trkA), brain-derived neurotrophic factor and neurotrophin-4/5 (trkB), and neurotrophin-3 (trkC). We examine the expression of these three neurotrophin tyrosine kinase receptors in the chick auditory system throughout most of development. Trks were localized in the auditory brainstem, the cochlear ganglion, and the basilar papilla of chicks from embryonic (E) day 5 to E21, by using antibodies and standard immunocytochemical methods. TrkB mRNA was localized in brainstem nuclei by in situ hybridization. TrkB and trkC are highly expressed in the embryonic auditory brainstem, and their patterns of expression are both spatially and temporally dynamic. During early brainstem development, trkB and trkC are localized in the neuronal cell bodies and in the surrounding neuropil of nucleus magnocellularis (NM) and nucleus laminaris (NL). During later development, trkC is expressed in the cell bodies of NM and NL, whereas trkB is expressed in the nerve calyces surrounding NM neurons and in the ventral, but not the dorsal, dendrites of NL. In the periphery, trkB and trkC are located in the cochlear ganglion neurons and in peripheral fibers innervating the basilar papilla and synapsing at the base of hair cells. The protracted expression of trks seen in our materials is consistent with the hypothesis that the neurotrophins/tyrosine kinase receptors play one or several roles in the development of auditory circuitry. In particular, the polarized expression of trkB in NL is coincident with refinement of NM terminal arborizations on NL.     

Bioactive tumour necrosis factor alpha but not granulocyte- macrophage colony-stimulating factor correlates inversely with Langerhans' cell numbers in skin tumours

Langerhans' cells (LCs) are thought to play an important role in presentation of tumour antigens for the induction of anti-tumour immunity. Epidermis overlying some transplanted murine skin tumours contains increased numbers of LCs; however, alterations in LC numbers are not related to tumour antigenicity or host immunity, suggesting that another factor(s), such as tumour-produced cytokines, influences LC density. It has been postulated that dendritic epidermal T cells (DETCs) play a role in immunosurveillance within the normal epidermis. Two cytokines which potentially alter LC numbers or function include granulocyte-macrophage colony-stimulating factor (GM-CSF) and tumour necrosis factor-α (TNF-α). GM-CSF maintains LC viability in culture, and there are reports that it can increase LC density. There is evidence that TNF-α induces LC to migrate from the epidermis. In the present study, LC densities in regressor and non-regressor murine skin tumours and overlying epidermis were enumerated, and bioactive GM-CSF and TNF-α present in the tumours were measured. We found significantly increased epidermal LC numbers above non-regressor, but not regressor, tumours. DETC numbers were significantly increased above some tumours. Although all tumour types produced TNF-α, the regressors, which did not increase LC numbers, produced the most TNF-α. In contrast, tumour production of GM-CSF did not correlate with any pattern of alteration of LC density or tumour growth. Tumour production of neither cytokine nor tumour growth correlated with DETC numbers overlying tumours. Our results suggest that TNF-α may be associated with skin tumour regression and may prevent LC accumulation by tumours. Int. J. Cancer 75:210–216, 1998. © 1998 Wiley-Liss, Inc.     

Nanocrystalline zirconia: A novel sorbent for the preparation of 188W/188Re generator

Nanocrystalline zirconia, a novel high capacity sorbent material was synthesized and tested for its utility in the preparation of ¹⁸⁸W/¹⁸⁸Re generators. The structural investigation of the material was carried out using X-ray diffraction, surface area determination, FTIR and TEM micrograph analysis. Various experimental parameters were optimized to separate ¹⁸⁸Re from ¹⁸⁸W. The capacity of the material was found to be ～325 mg W/g at the optimum pH. A chromatographic ¹⁸⁸W/¹⁸⁸Re generator was developed using this material from which >80% of ¹⁸⁸Re generated could be eluted with 0.9% saline solution, with high radionuclidic, radiochemical and chemical purity and appreciably high radioactive concentration suitable for radiopharmaceutical applications.     

Consensus guidelines for the management of atopic dermatitis: An Asia–Pacific perspective

Atopic dermatitis (AD) is a relatively common disease in patients in the Asia–Pacific region. It presents a particular clinical challenge and requires careful clinical management. The chronic nature of AD characterized by flares, exacerbations and periods of quiescence requires a multipronged approach aimed at reducing itch, inflammation and the appearance of secondary lesions. In addition, varying levels of maintenance therapy may be required to avoid exacerbations. Survey data from the region indicate that there is significant variation across the Asia–Pacific with regard to current treatment practices. The management of AD may also be influenced by differing health‐care systems, variable climate, access to medical care and cultural diversity. The current consensus guidelines have been developed to provide up‐to‐date and concise evidence‐ and experience‐based recommendations directed towards general practitioners and general dermatologists in the Asia–Pacific region on the management of pediatric and adult AD.