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Surface plasmon resonance analysis of antipolysaccharide antibody specificity: responses to meningococcal group C conjugate vaccines and bacteria.

Antibody (Ab) responses to polysaccharides (PS), such as Neisseria meningitidis group C PS (MCPS), are characterized as being thymus independent and are restricted with regard to clonotype and isotype expression. PS conjugated to proteins, e.g., MCPS coupled with tetanus toxoid or the diphtheria toxin derivative CRM197, elicit thymus-dependent responses. The present study developed a surface plasmon resonance approach to evaluate Ab responses to MCPS conjugate vaccines, including either O-acetylated (OAc+) or de-O-acetylated (OAc-) forms of the PS. The results were generally consistent with those obtained by enzyme-linked immunosorbent assay and showed that sera from mice immunized with conjugate vaccines contain Abs that bind more effectively to OAc+ and OAc- MCPS than sera from mice immunized with fixed bacteria. The data suggest a critical shared or overlapping epitope recognized by all the conjugate vaccine immune sera and strategies for assessing polyclonal Ab avidity.

Acetylation↗

Evolution and molecular characterization of methicillin-resistant Staphylococcus aureus epidemic and sporadic clones in Cordoba, Argentina.

Since 1999, a new, epidemic, methicillin-resistant Staphylococcus aureus (MRSA) strain, named the "Cordobes clone," has emerged in Argentina and coexists with the pandemic Brazilian clone. The purpose of this study was to determine the stability over time of the new clone and to investigate its evolutionary relationship with epidemic international MRSA lineages and with other MRSA and methicillin-susceptible S. aureus (MSSA) major clones distributed in this region. One hundred three MRSA isolates recovered in 2001 from Cordoba, Argentina, hospitals and 31 MSSA strains collected from 1999 to 2002 were analyzed by their antibiotic resistance patterns, phage typing, and pulsed-field gel electrophoresis. Additionally, representative members of most MRSA defined genotypes (A, B, C, E, K, and I) were characterized by multilocus sequence typing (MLST) and spaA and SCCmec typing. The most prevalent MSSA pulsotypes were also analyzed by MLST. Our results support the displacement of the Brazilian clone (sequence type [ST] 239, spaA type WGKAOMQ, SCCmec type IIIA) by the Cordobes clone (ST5, spaA type TIMEMDMGMGMK, SCCmec type I) in the hospital environment. MRSA and MSSA isolates shared only ST5. The data support the origin of the Cordobes clone as a member of a lineage that includes the pediatric and New York/Japan international clones and that is genetically related to the British EMRSA-3 strain. Interestingly, the pediatric clone, isolated from most community-acquired infections in Cordoba, was characterized by ST100, a single-locus variant of ST5 and a new variant of SCCmec type related to SCCmec type IVc.

Anti-Bacterial Agents↗

Beyond counting cases: public health impacts of national Paediatric Surveillance Units.

Paediatric Surveillance Units (PSUs) have been established in 14 countries and facilitate national, prospective, active surveillance for a range of conditions, with monthly reporting by child health specialists. The International Network of Paediatric Surveillance Units (INoPSU) was established in 1998 and facilitates international collaboration among member PSUs and allows for sharing of resources, simultaneous data collection and hence comparison of data from different geographical regions. The impact of data collected by PSUs, both individually and collectively as members of INoPSU, on public health outcomes, clinical care and research is described.

Child↗

Analysis of LMP and TAP polymorphisms by polymerase chain reaction-restriction fragment length polymorphism in rheumatoid arthritis.

OBJECTIVE: The aim of this study was to investigate the relation between the polymorphism of large molecular weight proteasome (LMP) (LMP2-LMP7) and transporter associated with antigen processing (TAP) (TAP1-TAP2) genes and rheumatoid arthritis (RA). METHODS: Sixty RA patients and 102 ethnically matched unrelated healthy subjects were typed for LMP, TAP, and disease associated HLA-DRB1 alleles by using a new strategy based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with amplification created restriction sites. RESULTS: The polymorphism of LMP (LMP2-LMP7) and TAP (TAP1-TAP2) genes was examined in shared epitope positive and negative RA patients and controls. No significant differences in the LMP or TAP allele frequencies were observed between the total patient and control groups or the patients and controls groups or the patients and controls positive or negative for the shared epitope. CONCLUSION: The data suggest that the polymorphisms of LMP and TAP genes do not have an important influence in the pathogenesis of RA, although larger studies will be needed to provide more conclusive evidence on the role of these genes in RA. A new, highly reliable strategy for typing LMP alleles is also described.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Receptor phosphorylation does not mediate cross talk between muscarinic M(3) and bradykinin B(2) receptors.

This study examined cross talk between phospholipase C-coupled muscarinic M(3) and bradykinin B(2) receptors coexpressed in Chinese hamster ovary (CHO) cells. Agonists of either receptor enhanced phosphoinositide signaling (which rapidly desensitized) and caused protein kinase C (PKC)-independent, homologous receptor phosphorylation. Muscarinic M(3) but not bradykinin B(2) receptors were also phosphorylated after phorbol ester activation of PKC. Consistent with this, muscarinic M(3) receptors were phosphorylated in a PKC-dependent fashion after bradykinin B(2) receptor activation, but muscarinic M(3) receptor activation did not influence bradykinin B(2) receptor phosphorylation. Despite heterologous phosphorylation of muscarinic M(3) receptors, phosphoinositide and Ca(2+) signaling were unaffected. In contrast, marked heterologous desensitization of bradykinin-mediated responses occurred despite no receptor phosphorylation. This desensitization was associated with a sustained component of muscarinic receptor-mediated signaling, whereas bradykinin's inability to influence muscarinic receptor-mediated responses was associated with rapid and full desensitization of bradykinin responses. Thus the mechanism of functional cross talk most likely involves depletion of a shared signaling component. These data demonstrate that receptor phosphorylation is not a prerequisite for heterologous desensitization and that such desensitization is not obligatory after heterologous receptor phosphorylation.

Animals↗

Widespread mechanosensory activation of the serotonergic system of the medicinal leech.

The serotonergic system of the medicinal leech comprises a small number of iterated, identified neurons, of which the Retzius (Rz) neurons are major components. Activity in pressure mechanosensory (P) cells sufficient to elicit locomotory and defensive behaviors also excites Rz neurons. We characterized the interactions between P and Rz neurons within the ganglion and at different distances along the nerve cord. 2. Within a ganglion 1) P cells excited both Rz neurons, electrically close to the site of electrical coupling between the Rz neurons; 2) each of the four P cells had similar effects on the Rz neurons; and 3) homologous contralateral P cells shared interneuronal pathways. These data show that P cells provide nearly identical bilateral information onto Rz neurons. 3. Along the nerve cord 1) every P cell excited Rz neurons in ganglia anterior and posterior to the site of stimulation; 2) the signal was carried the entire length of the nerve cord along interneuronal pathways with similar overall (but regionally different) conduction velocities in the two directions; 3) the amplitude of the Rz responses was smaller as the distance to the activated P cell increased; 4) the rate of change of the amplitude along the cord was larger when the signal traveled from front-to-back than in the opposite direction. 4. These data shows that mechanosensory input from any segment could excite Rz neurons along the cord, in proportion to the intensity of the stimulus.

Animals↗

Cranial nerves of the coelacanth, Latimeria chalumnae [Osteichthyes: Sarcopterygii: Actinistia], and comparisons with other craniata.

We reconstructed the cranial nerves of a serially sectioned prenatal coelacanth, Latimeria chalumnae. This allowed us to correct several mistakes in the literature and to make broad phylogenetic comparisons with other craniates. The genera surveyed in our phylogenetic analysis were Eptatretus, Myxine, Petromyzon, Lampetra, Chimaera, Hydrolagus, Squalus, Mustelus, Polypterus, Acipenser, Lepisosteus, Amia, Neoceratodus, Protopterus, Lepidosiren, Latimeria and Ambystoma. Cladistic analysis of our data shows that Latimeria shares with Ambystoma two characters of the cranial nerves. Our chief findings are: 1) Latimeria possesses an external nasal papilla and pedunculated olfactory bulbs but lacks a discrete terminal nerve. In other respects its olfactory system resembles the plesiomorphic pattern for craniates. 2) The optic nerve is plicated, a character found in many but not all gnathostomes. Latimeria retains an interdigitated partial decussation of the optic nerves, a character found in all craniates surveyed. 3) The oculomotor nerve supplies the same extrinsic eye muscles as in lampreys and gnathostomes. As in gnathostomes generally, Latimeria has a ciliary ganglion but its cells are located intracranially in the root of the oculomotor nerve, and their processes reach the eye via oculomotor and profundal rami. 4) The trochlear nerve supplies the superior oblique muscle as in all craniates that have not secondarily reduced the eye and its extrinsic musculature. 5) The profundal ganglion and ramus are entirely separate from the trigeminal system, with no exchange of fibers. This character has an interesting phylogenetic distribution: in hagfishes, lampreys, lungfishes and tetrapods, the profundal and trigeminal ganglia are fused, whereas in other taxa surveyed the ganglia are separate. The principal tissues innervated by the profundal nerve are the membranous walls of the tubes of the rostral organ. 6) As in lampreys and gnathostomes, the trigeminal nerve has maxillary and mandibular rami. Unlike all other gnathostomes surveyed, the trigeminal nerve of Latimeria lacks a sizable superficial ophthalmic ramus. Thus, Latimeria lacks the well-developed superficial ophthalmic complex reported in most other fishes. As in gnathostomes generally, the maxillary ramus of the trigeminal nerve fuses with the buccal ramus of the anterodorsal lateral line nerve to form the buccal+maxillary complex. We reject the term 'Gasserian ganglion', which is often applied to the fused profundal and trigeminal ganglion of tetrapods. 7) The abducent nerve innervates not only the lateral rectus muscle (a character common to myopterygians) but also the basicranial muscle. As we previously reported, it is probable that the basicranial muscle of Latimeria is homologous to the ocular retracter muscle of amphibians.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Comparison of aqueous grain dusts, cotton dusts, and mold extracts by crossed immunoelectrophoretic techniques.

To evaluate the possibility that an immune response is involved in the pathogenesis of grain dust or cotton dust induced respiratory disease, 1 grain dust extract (A) was compared with 4 similar grain dust extracts, a cotton dust extract, and a mixture of fungal preparations, using crossed immunoelectrophoretic techniques and rabbit anti-A sera. All extracts demonstrated a significant number of shared antigenic determinants. These data suggest that grain and cotton dust may induce disease via a common pathogenic mechanism and putative agent(s).

Allergens↗

Lessons from European population genetic databases: comparing the law in Estonia, Iceland, Sweden and the United Kingdom.

The advent of large-scale, population genetic databases (PGDs) in several countries around the world marks a significant development in human DNA banking and genetic research. The European countries that have led the way in the development of PGDs are Iceland, Sweden, Estonia and the U.K. In legal terms, the emergence of PGDs has been far from straightforward as such projects pose a range of difficult and complex issues for the law to address. This article canvasses the current law in Iceland, Estonia, Sweden and the U.K. on four fundamental issues of principle pertaining to PGDs, in order to illustrate the difficulties that have emerged around PGDs, highlight key areas of legal concern, and shed light on possible ways forward. It compares and contrasts the differing legal positions and lawmakers' responses to date in these four European countries that have established PGDs or are seeking to do so. The four fundamental issues examined are: (1) consent, especially for secondary research purposes; (2) ownership of biological samples, data and databases; (3) the rights of certain third parties to gain access to, and to use, PGD biological samples and data; and (4) benefit sharing, including the provision of feedback and genetic counselling to participants. This analysis may offer some guidance for policymakers in other jurisdictions where PGDs have been proposed or are being established.

Databases, Genetic↗

Psychiatric residences: notification, NIMBY, and neighborhood relations.

OBJECTIVE: Federal legislation has given mental health agencies the authority to forego notifying neighbors when establishing shared, supervised houses and apartments for individuals with severe mental illness, but many continue to notify neighbors in the belief that outreach enhances community integration. This study examined the associations between notification strategies and initial neighborhood opposition and longer-term neighborhood relations. METHODS: Telephone interviews were conducted with 169 mental health administrators in seven states who described the notification strategy and short- and long-term neighborhood responses to their most recently established shared, supervised residence. Additional data on neighborhood relations were collected in interviews with 138 staff who worked on site at the residences described by the mental health administrators. RESULTS: Just under half of the agencies used a strategy that involved notification of neighbors, and those organizations were significantly more likely to experience initial opposition (59% of notifiers experienced opposition compared with 35% of non-notifiers). Those who notified neighbors were more likely to initiate activities such as an open house with neighbors after the site was established. There were no significant associations between current neighborhood attitudes and initial notification strategies or initial opposition. Agencies that initiated postsiting social activities with neighbors were significantly more likely to report "very accepting" attitudes. CONCLUSIONS: Administrators who use notification may experience higher levels of initial opposition, but longer-term neighborhood relations may not vary significantly regardless of whether neighbors were notified or whether initial opposition was present. Some agency-sponsored postsiting activities may facilitate substantive connections with neighbors.

Attitude↗

The Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM): a review of the ICCVAM test method evaluation process and current international collaborations with the European Centre for the Validation of Alternative Methods (ECVAM).

Over the last decade, national authorities in the USA and Europe have launched initiatives to validate new and improved toxicological test methods. In the USA, the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) and its supporting National Toxicology Program Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM) were established by the Federal Government to work with test developers and Federal agencies to facilitate the validation, review, and adoption of new scientifically sound test methods, including alternatives that can refine, reduce, and replace animal use. In Europe, the European Centre for the Validation of Alternative Methods (ECVAM) was established to conduct validation studies on alternative test methods. Despite differences in organisational structure and processes, both organisations seek to achieve the adoption and use of alternative test methods. Accordingly, both have adopted similar validation and regulatory acceptance criteria. Collaborations and processes have also evolved to facilitate the international adoption of new test methods recommended by ECVAM and ICCVAM. These collaborations involve the sharing of expertise and data for test-method workshops and independent scientific peer reviews, and the adoption of processes to expedite the consideration of test methods already reviewed by the other organisation. More recently, NICEATM and ECVAM initiated a joint international validation study on in vitro methods for assessing acute systemic toxicity. These collaborations are expected to contribute to accelerated international adoption of harmonised new test methods that will support improved public health and provide for reduced and more-humane use of laboratory animals.

Animal Testing Alternatives↗

Clinical quality improvement in a multihospital system: the voluntary hospitals of America/Pennsylvania experience.

Sharing of patient outcome data among hospitals hastened the evolution of continuous quality improvement at Voluntary Hospitals of American/Pennsylvania (VHA/PA) to a focus on learning from the best: clinical benchmarking. Analysis of VHA/PA MedisGroups comparative outcome data showed significant variation. In the first study, variations from 3 to 25% normal appendectomy rates were noted. A second study on primary and secondary cesarean section rates again showed variation in outcomes. A study of the benchmark hospitals showed characteristics, specific to those institutions, that resulted in excellent outcomes. To quantify best processes, VHA/PA has embarked on a clinical benchmarking project with the assistance of MediQual Systems, Inc.

Adolescent↗

The barriers to social inclusion as perceived by people with intellectual disabilities.

Very little research has been done on social inclusion from the perspective of people with intellectual disabilities, including perceived barriers and remedies. Focus groups were held with 68 persons, mostly tenants in supported living or shared group homes. Qualitative data were obtained using a mix of workshop activities and small group discussions. Four main barriers were identified: lack of necessary knowledge and skills; role of support staff and service managers; location of house; and community factors such as lack of amenities and attitudes. Participants were able to identify a range of solutions for these barriers. Most of their proposals were in line with the aims of current government policy and good practice. These findings reinforce the contribution individuals could make to the planning of local services for themselves and others. This advocacy has been an essential element in reducing the social isolation of other marginalized groups.

Adult↗

An Asilomar moment.

As governments in the US and Europe contemplate legislation that would divert funding of some genomics-driven research to Offices of Homeland Security and the like, and that would restrict the freedom of biologists to publish and share some of their data, we of the scientific community are facing a crisis, brought on by fears of bioterrorism, that eerily mirrors the early days of recombinant DNA research.

Access to Information↗

Differentiation of HL-60 cells into cells with the osteoclast phenotype.

The osteoclast is the unique multinucleated cell that is responsible for bone degradation in both physiological and pathological circumstances. However, knowledge of the lineage of this inaccessible cell, the nature of its precursors, and the regulation of its formation and activation is limited and controversial. Here we show that the human promyelocytic cell line HL-60 has the potential, under the appropriate culture conditions, to differentiate into cells that have morphological and functional characteristics of osteoclasts, including multinucleation, presence of tartrate-resistant acid phosphatase activity, cross-reactivity with monoclonal antibodies that preferentially recognize osteoclasts, and capacity to resorb bone and respond to calcitonin. The multinucleated cells also contain high affinity receptors to calcitonin, in contrast to wild-type undifferentiated HL-60 cells. These data suggest that osteoclasts share a common precursor with hematopoietic cells. These undifferentiated and differentiated HL-60 cells should provide a unique model for study of the cell biology of human osteoclast differentiation, allowing molecular biological and biochemical studies heretofore not possible.

Bone Resorption↗

Although DR3-DQB1*0201 may be associated with multiple component diseases of the autoimmune polyglandular syndromes, the human leukocyte antigen DR4-DQB1*0302 haplotype is implicated only in beta-cell autoimmunity.

Human leukocyte antigen (HLA)-DRB1 and -DQB1 alleles were analyzed using a PCR-based sequence-specific priming technique in 16 patients with autoimmune polyglandular syndrome type I (APS-I), 31 patients with APS-II, and 110 patients with component diseases of APS-II, including 9 patients with isolated Addison's disease, 43 patients with Hashimoto's thyroiditis, 22 patients with Graves' disease, and 36 patients with vitiligo. No significant associations was observed between HLA and APS-I patients in our data set, nor was sharing of HLA haplotypes by sibling pairs affected by APS I significantly different from the random expectation. Thus, HLA-DRB1 and -DQB1 genes are probably not involved in APS-I. To delineate the associations between HLA-DRB1, DQB1, and APS-II, we analyzed APS-II patients with or without beta-cell autoimmunity [i.e. insulin-dependent diabetes (IDD) and/or islet cell or glutamic acid decarboxylase autoantibodies]. Our results suggest that the association between DR4-DQB1*0302 and APS-II was entirely due to the presence of pancreatic beta-cell autoimmunity, since this haplotype was otherwise not significantly associated with APS-II or with any other of its component diseases. In contrast, the DR3-DQB1*0201 haplotype was associated not only with IDD, but also with APS-II in the absence of pancreatic beta-cell autoimmunity, as were several its component diseases, including isolated Addison's disease, Graves' disease, and Hashimoto's thyroiditis. Interestingly, the frequency of DQB1*0602, a dominantly protective allele against IDD, was not significantly decreased in the APS-II patients with IDD or beta-cell autoimmunity, albeit the patient numbers were small. This phenomenon may suggest that the development of autoimmunity to nonpancreatic endocrine glands may predispose autoimmunity to the pancreatic beta-cells and involve genes other than those of the MHC.

Adolescent↗

Social and cognitive processes in interpersonal communication: implications for advanced telecommunications technologies.

Interactive multimedia conferencing systems, in which two or more remotely located people can work on cooperative tasks through shared audio, video, and data, appear to be the wave of the future. However, because of great advances in the underlying technology of multimedia conferencing systems, many design decisions have been driven by what is technically feasible as opposed to what will best suit the needs of the users. In this paper we provide a framework for the design and evaluation of features in advanced telecommunications products and services which is derived from empirical research on interpersonal communication. We also discuss implications of this research for the development and use of advanced telecommunications technologies.

Cognition↗

Influenza and other respiratory virus-related emergency department visits among young children.

BACKGROUND: Influenza and other winter respiratory viruses cause substantial morbidity among children. Previous estimates of the burden of illness of these viruses have neglected to include the emergency department, where a large number of patients seek acute care for respiratory illnesses. This study provides city- and statewide population estimates of the burden of illness attributable to respiratory viruses for children receiving emergency department-based care for respiratory infections during the winter months. METHODS: The number of patients < or = 7 years of age presenting to the emergency department of an urban tertiary care pediatric hospital with acute respiratory infections was estimated by using a classifier based on presenting complaints. The rates of specific viral infections in this population were estimated by using the rates of positivity for respiratory syncytial virus, influenza virus, parainfluenza virus, adenovirus, and enterovirus. Local emergency department market share and US Census data enabled determination of the rates of emergency department visits in the Boston, Massachusetts, area and in Massachusetts. RESULTS: During the 11-year study period, the mean yearly number of patients < or = 7 years of age presenting to the study emergency department during the winter season was 17397. On the basis of the respiratory classifier, the mean number of patients with an acute respiratory infection was 6923, or 398 per 1000 emergency department visits. In the city population, the mean number of emergency department visits for acute respiratory infections was 17906, which is equivalent to 113.9 per 1000 children residing in the city, and in the state population the mean number was 61529, or 94.5 per 1000 children residing in the state. At the state level, 23114 of the visits were for respiratory syncytial virus, 5650 for influenza, 1751 for parainfluenza virus, 2848 for adenovirus, and 798 for enterovirus. For patients 6 to 23 months of age in the state population, there were 19860 emergency department visits for acute respiratory infections, or 168 per 1000 children in this age group, with 6235 visits resulting from respiratory syncytial virus and 2112 resulting from influenza. CONCLUSION: There is a high incidence of emergency department visits for infectious respiratory illnesses among children. This important component of health care use should be included in estimates of the burden of illness attributable to influenza and other winter respiratory viruses.

Boston↗