Mother to child transfer of IgG and IgA antibodies against Dermatophagoides pteronyssinus.

There is strong evidence from animal models that placental and/or breast milk-mediated transfer of maternal allergen-specific IgG prevents allergic immune responses in the progeny. Both human and animal data also point to IgA as having an important regulatory role. In contrast, little is known about maternal transfer of IgG and IgA specific for respiratory allergens in humans. Dermatophagoides pteronyssinus (Der p) is an indoor allergen that is a major cause of asthma worldwide. We analysed maternal to child Der p-specific IgG and IgA transfer in a cohort of 77 paired maternal and child samples. We found Der p-specific IgG and its IgG1, IgG2 and IgG4 subclasses in all cord blood samples. Except for IgG1, cord levels were higher in newborns from atopic mothers (n = 29) compared to non-atopic mothers (n = 48). Der p-specific IgA was found in all colostrum samples and levels were independent of maternal atopic status. Notably, anti-Der p IgG was also found in colostrum and levels were higher in atopic mothers. We believe that our work is a critical first step in the identification of early factors that may impact asthma development and should guide the development of clinical studies that assess whether Der p-specific IgG and IgA protect children from allergy as demonstrated in animal models.

Evaluation of the immune humoral response of Brazilian patients with Rubinstein-Taybi syndrome

Rubinstein-Taybi syndrome (RTS) is a rare developmental disorder characterized by craniofacial dysmorphisms, broad thumbs and toes, mental and growth deficiency, and recurrent respiratory infections. RTS has been associated with CREBBP gene mutations, but EP300 gene mutations have recently been reported in 6 individuals. In the present study, the humoral immune response in 16 RTS patients with recurrent respiratory infections of possible bacterial etiology was evaluated. No significant differences between patients and 16 healthy controls were detected to explain the high susceptibility to respiratory infections: normal or elevated serum immunoglobulin levels, normal salivary IgA levels, and a good antibody response to both polysaccharide and protein antigens were observed. However, most patients presented high serum IgM levels, a high number of total B cell and B subsets, and also high percentiles of apoptosis, suggesting that they could present B dysregulation. The CREBBP/p300 family gene is extremely important for B-cell regulation, and RTS may represent an interesting human model for studying the molecular mechanisms involved in B-cell development.

Evaluation of the immune humoral response of Brazilian patients with Rubinstein-Taybi syndrome.

Rubinstein-Taybi syndrome (RTS) is a rare developmental disorder characterized by craniofacial dysmorphisms, broad thumbs and toes, mental and growth deficiency, and recurrent respiratory infections. RTS has been associated with CREBBP gene mutations, but EP300 gene mutations have recently been reported in 6 individuals. In the present study, the humoral immune response in 16 RTS patients with recurrent respiratory infections of possible bacterial etiology was evaluated. No significant differences between patients and 16 healthy controls were detected to explain the high susceptibility to respiratory infections: normal or elevated serum immunoglobulin levels, normal salivary IgA levels, and a good antibody response to both polysaccharide and protein antigens were observed. However, most patients presented high serum IgM levels, a high number of total B cell and B subsets, and also high percentiles of apoptosis, suggesting that they could present B dysregulation. The CREBBP/p300 family gene is extremely important for B-cell regulation, and RTS may represent an interesting human model for studying the molecular mechanisms involved in B-cell development.

IL-10 and TGF-beta immunoregulatory cytokines rather than natural regulatory T cells are associated with the resolution phase of Vogt-Koyanagi-Harada (VKH) syndrome.

The pro-inflammatory cytokines play a critical role in the initiation and propagation of ocular autoimmune diseases. Regulation of these cytokines is generally mediated by the immunoregulatory cytokine such as IL-10 or TGF-beta. In this study, we investigated the immunoregulatory cytokine profile and frequency of natural regulatory T cells (nTregs) in patients with Vogt-Koyanagi-Harada (VKH). We obtained the peripheral blood mononuclear cells (PBMC) from patients with VKH and healthy controls. The cytokine profile from supernatants of PBMC cultured with or without phytohaemagglutinin (PHA) was measured by ELISA, the percentage of CD4(+) Foxp3(+) and CD25(high)Foxp3(+) T regulatory cells were analysed by flow cytometry, and the transcriptional level of Foxp3 expression was analysed by real-time quantitative PCR. The immunoregulatory cytokines, TGF-beta and IL-10, increased in patients with VKH in the inactive stage of the disease. We observed no significant difference in the CD4(+) Foxp3(+) and CD25(high)Foxp3(+) T cells as well as no reduction in FOXP3 mRNA expression in the patients with VKH when compared to healthy controls. We showed in our work, an increase in IFN-gamma secretion by PBMC of patients with VKH in the active stage of the disease when compared to healthy controls and patients in the inactive stage. Our data suggest that IL-10 and TGF-beta cytokines, rather than nTregs are associated with the resolution phase of the disease and may have a more relevant role in controlling this disease.

Human IgG but not IgM antibodies can protect mice from the challenge with live O6 Escherichia coli.

We evaluated the ability of human anti-lipopolysaccharide (LPS) O6 immunoglobulin G (IgG) and IgM antibodies to protect mice challenged with Escherichia coli serotype O6:K2ac. Purified whole IgG, commercial gammaglobulin, whole IgM-effluent, pool of normal human serum (NHS), agammaglobulinaemic serum (test groups) or phosphate-buffered saline (control group) was injected into adult male 18 h before a challenge with viable O6 E. coli. The mortality rate was assessed over a period of 72 h. To determine the opsonic and phagocytic activity of the antibody isotypes, we incubated peritoneal macrophages from the control and test groups collected at different times after challenge with the live bacteria with acridine orange for fluorescent analysis. Tumour necrosis factor (TNF)-alpha and interleukin (IL)-6 were quantified in serum of both the test and control groups. All mice that received commercial gammaglobulin or NHS survived. Purified whole IgG (containing 1.1 mg/l of anti-LPS O6 IgG antibodies) protected 87.5% of the animals tested in this experiment, while whole IgM-enriched effluent with 1.5 mg/l of anti-LPS O6 IgM antibodies protected only 12.5%. The agamma serum showed no protective capacity compared with PBS (serving as control). The minimal concentration of anti-LPS O6 IgG antibodies able to protect 50% of animals was 0.137 mg/l of purified whole IgG. Whole IgM-enriched effluent showed no protective capacity independently of the concentration tested (0.048-17.0 mg/l of anti-LPS O6 IgM antibodies). Fluorescent analysis of peritoneal macrophages from animals pretreated with purified whole IgG showed no bacteria at 8 h after the challenge. By contrast, whole IgM effluent showed an increasing number of live bacteria at the same time. Mice that had received whole IgM effluent (1.5 mg/l of anti-LPS O6 IgM antibodies) before the challenge with LPS O6 presented 20.5 microg/l of IL-6 and 1.5 microg/l of TNF-alpha. Serum from animals pretreated with purified IgG did not present any detectable pro-inflammatory cytokine. Our findings suggest that IgG but not IgM antibodies protect animals from a challenge with E. coli O6 serotype.

Salivary lysozyme levels in patients with primary immunodeficiencies.

BACKGROUND: Lysozyme is a muramidase that acts on the peptideoglycan wall of Gram positive bacteria, causing cell death. It plays part in innate immunity and is present in blood, external fluid, as well in lysossomal granules of the phagocytes. Primary Immunodeficiencies are a diverse group of illnesses that, as a result of abnormalities of the immune system, increase susceptibility to infection. Among the examples of impaired natural immunity are defects in phagocytes and in the complement system. Innate immunity could be important in protecting mucosas against infections in patients with different forms of primary immunodeficiencies. The aim of this study was to investigate lysozyme concentrations in saliva from patients with primary immunodeficiencies. METHODS: Lysozyme levels in saliva samples from 34 patients with primary immunodeficiency (30 children and adolescents between the age of 3-13 years and 4 adults between the age of 20-33) and 60 age-matched healthy controls (49 children and adolescents between the ages of 3-15 and 11 adults between the ages of 22-42) were determined by the lysoplate method. RESULTS: There was no statistically significant difference between the lysozyme concentrations in the saliva of the immunodeficient subjects and those of the healthy controls. CONCLUSION: The results in the present work clearly show that salivary lysozyme levels in primary immunodeficient patients are equivalent to those found in healthy controls, suggesting that this enzyme still represents a remaining (but not a compensatory mechanism), contributing to the protection of there patients against infections.

Salivary lysozyme levels in patients with primary

Background: Lysozyme is a muramidase that acts on the peptideoglycan wall of Gram positive bacteria, causing cell death. It plays part in innate immunity and is present in blood, external fluid, as well in Iysossomal granules of the phagocytes. Primary Immunodeficiencies are a diverse group of iIInesses that, as a result of abnormalities of the immune system, increase susceptibility to infection. Among the examples of impaired natural immunity are defects in phagocytes and in the complement system. Innate immunity could be important in protecting mucosas against infections in patients with different forms of primary immunodeficiencies. The aim of this study was to investigate Iysozyme concentrations in saliva from patients with primary immunodeficiencies. Methods: Lysozyme levels in saliva samples from 34 patients with primary immunodeficiency (30 children and adolescents between the age of 3-13 years and 4 adults between the age of 20-33) and 60 agematched healthy controls (49 children and adolescents between the ages of 3-15 and 11 adults between the ages of 22-42) were determined by theIysoplate method. Results: There was no statistically significant difference between the Iysozyme concentrations in the saliva of the immunodeficient subjects and those of the healthy controls. Conclusion: The results in the present work clearly show that salivary Iysozyme levels in primary immunodeficient patients are equivalent to those found in healthy controls, suggesting that this enzyme still represents a remaining (but not a compensatory mechanism), contributing to the protection of there patients against infections

Introducción: La lisozima es una muramidasa que actua sobre la pared de peptidoglicano de las bacterias Gram-positivas, provocando la muerte celular. Desempeña un papel en la inmunidad innata y está presente en la sangre, secreciones externas y en los gránulos de los fagocitos. Las inmunodeficiencias primarias constituyen un grupo diverso de enfermedades que, como consecuencia de las anormalidades en el sistema inmune, presentan un aumento de susceptibilidad a las infecciones. Ejemplos de deficiencias de la inmunidad natural son los defectos de los fagocitos y del sistema de complemento. La inmunidadinnata puede ser importante para la protección contra las infecciones de las mucosas en los pacientes con diferentes tipos de inmunodeficiencia primaria. El objetivo de este estudio es investigar la concentración de lisozima en la saliva en pacientes con inmunodeficiencia primaria. Métodos: Los niveles de lisozima en las muestras de saliva de 34 pacientes con inmunodeficiencia primaria (30 niños y adolecentes con edad entre 3-13 años y 4 adultos con edad entre 20-33 años) y 60 controles sanos de la misma edad (49 niños y adolescentes con edad entre 3-15 años y 11 adultos conedad entre 22-42 años) fueron determinados por el método Iysoplate. Resultados: No se observó una diferencia estadística significativa entre las concentraciones de lisozima en la saliva de los individuos inmunodeficientesy de los controles sanos. Conclusión: Los resultados del presente trabajo claramente indican que los niveles de lisozima en la saliva de los pacientes con inmunodeficiencias primarias son equivalentes a los de los controles sanos, sugiriendo que este enzima representa un mecanismo remanescente, pero no compensador, que contribuye a la protección contra las infecciones en estos pacientes

Elevated levels and different repertoire profile of colostral anti-LPS antibodies may have a significant role in compensating newborn immunity.

A high prevalence of systemic infections caused by enterobacteria such as Escherichia coli is observed during the neonatal period. Lipopolysaccharide (LPS) is one of the major factors responsible for septic shock caused by these Gram-negative bacteria. We have recently demonstrated the presence of anti-LPS immunoglobulin (Ig)G antibodies in cord blood with a repertoire identical to that found in maternal serum. In the present study, we analyzed anti-LPS O111 antibody isotypes in maternal serum and colostrum from mothers and in cord serum from their respective full-term (n = 30) and preterm (n = 13) neonate infants. The main isotype found in serum samples from mothers of term infants was IgM (range between 28 and 54 mg/l), followed by IgA (1-2 mg/l) and IgG (2-3 mg/l). The range of IgG antibody concentrations in cord blood was between 2 and 3 mg/l, as a result of placental transfer. A novel observation in our study was that the LPS bands recognized by colostral antibodies were completely different from those recognized by IgG in serum. Colostral IgA antibodies recognized several bands not bound by serum IgG antibodies from the respective maternal serum, independently of the antibody quantity. In addition, we verified the pattern of LPS recognition by serum IgA and colostral IgA antibodies was identical, what suggested that the antibody isotype found in serum could probably be derived from differentiated IgA-positive cells which were homing to the mucosa through the mucosal homing mechanism. Identical pattern of recognition was obtained comparing the IgA and IgM isotypes in colostrum. Slight differences in the pattern of recognition were found between colostral and serum IgM antibodies. The fact that colostral antibodies recognize much more bands than serum antibodies may be important for the host to mount an effective immune response in the intestinal lumen, in order to prevent excessive absorption of LPS, reducing possible systemic effects caused by the molecule.

Developing an orthopaedic outcomes program.

Over the past several years there has been an increased effort to document outcomes, especially in the area of orthopaedics. Outcomes research is important because it can quantitatively and qualitatively demonstrate changes in a patient's condition, provide validation of clinical procedures, and impact upon patient satisfaction. Outcomes research is similar to traditional research in that it looks at a group of patients or a patient care process and focuses on the outcomes of treatment or care. An outcomes program is the infrastructure necessary to collect and analyze outcomes data within a scientifically sound framework. The steps of developing an outcomes program include identifying key players in the process, identifying the diagnoses and procedures to track in the program, deciding outcomes to measure, identifying instruments to be used, developing the data collection process, establishing a timeline, and developing the data entry and analysis process. A successful outcomes program is the product of a collaborative team effort. With knowledge of the necessary components and effective teamwork, it is relatively simple to establish a successful outcomes program.

Introduction to the nursing research process.

Many nurses have expressed an interest in conducting a nursing research project, but some have not had sufficient or recent exposure to the process of how to develop a research study. The nursing research process can be outlined using the nursing process steps of assessment, planning, intervention and evaluation. In the assessment phase the problem is identified, the literature is reviewed, and the variables are identified. In the planning phase the research question or hypothesis is formulated, and decisions are made on how the variables will be measured and how the sample will be chosen. In the intervention phase the data collection occurs; in the evaluation phase the data is analyzed and interpreted and the findings are communicated. By giving nurses a detailed yet understandable plan on how to conduct nursing research, their curiosity is encouraged and the body of knowledge will grow.

Creating an effective poster presentation.

One way to build knowledge in nursing is to share research findings or clinical program outcomes. The dissemination of these findings is often a difficult final step in a project that has taken months or years to complete. One method of sharing findings in a relaxed and informal setting is a poster presentation. This method is an effective form for presenting findings using an interactive approach. The milieu of a poster presentation enables the presenters to interact and dialogue with colleagues. Guidelines for size and format require that the poster is clear and informative. Application of design helps to create visually appealing posters. This article summarizes elements of designing and conducting a poster presentation.

Computer assisted outcomes research in orthopedics: total joint replacement.

Long-term studies are needed to determine clinically relevant outcomes within the practice of orthopedic surgery. Historically, the patient's subjective feelings of quality of life have been largely ignored. However, there has been a strong movement toward measuring perceived quality of life through such instruments as the SF-36. In a large database from an orthopedic practice results are presented. First, computerized data entry using touch screen technology is not only cost effective but user friendly. Second, patients undergoing hip or knee arthroplasty surgeries make statistically significant improvements in seven of the eight domains of the SF-36 in the first 3 months after surgery. Additional statistically significant improvements over the next 6 to 12 months are also seen. The data are presented here in detail to demonstrate the benefits of a patient outcomes program, to enhance the understanding and use of outcomes data and to encourage further work in outcomes measurement in orthopedics.

Evaluation of serum levels of IgG subclasses and anti-ribosyl-ribitolphosphate IgG and IgG2 in children with Haemophilus influenzae b meningitis.

In 40 children with Haemophilus influenzae b (Hib) meningitis, we determined serum levels (mg/dl) of IgG subclasses using the radial immunodiffusion method; 67.8 per cent of these children were less than 24 months old. In 14 children of the sample we measured serum IgG and IgG2 anti-ribosyl-ribitolphosphate (anti-PRP) (by enzyme-linked immunosorbent assay, ELISA) in the acute and convalescent phases of the disease. Lower IgG2 levels than those of the control group were obtained in all age ranges: 3-12 months, 1-2 years (p < 0.01), and 2-5 years (p < 0.001). IgG4 was also present in lower levels in patients of all age ranges (p < 0.05, p < 0.001, and p < 0.01 respectively). Serum levels of IgG anti-PRP and IgG2 anti-PRP measured were very low in the acute phase of the disease in all age ranges and there was no notable increase in levels during the convalescent phase of the disease. This result indicates that children less than 24 months old do not produce sufficient levels of IgG and IgG2 anti-PRP even after Hib meningitis.

Placental transfer of IgG antibodies against Haemophilus influenzae type b capsular polysaccharide in Brazilian term and preterm newborns.

Placental transfer of antibodies to polysaccharide antigens is still a controversial subject. The incidence of invasive Haemophilus influenzae type b (Hib) infections is high in countries where the vaccine has not been included in routine immunization schedules. In the present work, we proposed to evaluate the natural immune response to Hib capsular polysaccharide in term and preterm Brazilian newborns and their respective mothers. Although the means, medians, and ranges of antibody titres in paired maternal and cord sera from preterm neonates were similar, the maternal levels were slightly higher than the cord levels and a poor correlation between these levels was verified. Term neonates showed similar antibody levels to those of their respective mothers and a very significant correlation between these levels was observed.

Orthopaedic nursing research priorities: a replication and extension.

PURPOSE: This study identified current orthopaedic nursing research priorities that should be investigated to advance the practice of orthopaedic nursing. The study was accomplished by the National Association of Orthopaedic Nurses' (NAON) Research Committee via a national survey of selected NAON members. DESIGN: A descriptive design was used to determine research priorities. SAMPLE: The sample consisted of a random sample of 133 NAON members. The sample was stratified for either graduate degrees (> or = master's), other than a graduate degree (< or = bachelor's) (to insure representation from "frontline" practicing nurses), and registrants in the NAON Researcher Database and recipients of NAON Foundation or American Academy of Orthopaedic Surgeons (AAOS)/NAON grants. METHOD: A three round Delphi survey technique was used to build consensus by systematically generating, synthesizing, and analyzing opinions of a group of experts while maintaining confidentiality of the individuals. MAIN RESEARCH CLASSIFICATIONS: Nursing research priorities, Delphi method, Orthopaedic nursing. FINDINGS: The nine target research questions for orthopaedic nursing identified as high priority were grouped into the following categories: patient acuity, care delivery models, staffing issues, patient complications, pain management (in the elderly and those with altered mental status), and patient mobility. These research priority items are intended to direct the orthopaedic nurse researcher to study specific questions within these categories. CONCLUSION: Results reflect the dramatic changes occurring in orthopaedic nursing practice. Research priorities reveal the need for more research on pain and patient complications (e.g., deep vein thrombosis (DVT)) despite a preponderance of existing, published research on these topics. IMPLICATIONS FOR NURSING RESEARCH: This study identified target research questions for orthopaedic nursing. These questions may be used by orthopaedic nurses to develop nursing research proposals as well as collaborative research endeavors with other members of the orthopaedic health care team. An ongoing and wider dissemination of results of existing research to the NAON membership needs to be implemented.

[Immunological behavior (IgG, IgM, IgA) and total complement (CH50) of newborns infants with risk factors for early onset sepsis. Comparative analysis of newborns with and without infection]. / Comportamento imunológico (IgG, IgM, IgA) e complemento total (CH 50) de recém-nascidos com fatores de risco para sepse precoce. Análise comparativa entre recém-nascidos com e sem infecção.

Immunological behavior (IgG, IgM, IgA) and total Complement (CH50) of newborns infants with risk factors for early onset sepsis. Comparative analysis between newborns with and without infection. Rev. Hosp. Clín. Fac. Med. S. Paulo, 53(6): 303-310, 1998. The objective of this study was to verify the immunological behavior of the newborn infant in front of an infection. We studied 60 newborn infants that had risk factors for early onset sepsis (premature rupture membranes, clinic amnionitis or tract urinary infection) from de immunological and infection point of view. They were classified into three gestational age groups: < 34 weeks, between 34 and 36 6/7 weeks and > or = 37 weeks. Sepsis diagnosis was done through clinical and laboratorial data and we also included the followings exams: Immunological types (IgG, IgM, IgA) and total complement (CH50) obtained from the newborn at birth and on the fifth day of life. We could verify that 15 newborns (25%) presented early sepsis. There was a statistical association between perinatal asfixia and infection in the group with gestational age < 34 weeks and this same group presented statistical association between infection and death. The serical levels of IgG and CH50 were directly related to the gestational age and there were significant statistical differences between levels of IgG, IgM and total Complement between infected and not infected newborns within the same group os gestional age. We observed that the infection was associated to low levels of IgG and CH50, at birth and on the fifth day, mainly in the group of infected newborns with gestional age < 34 weeks, being this group, therefore, the one that would mostly benefit from an immunological support in front of and infection.

Transfer of IgG subclasses across placenta in term and preterm newborns.

In order to study placental transfer of IgG subclasses, paired blood samples were collected from mothers and umbilical cord of preterm (N = 69) and full-term (N = 68) newborns. The full-term group was further divided into 3 subgroups: appropriate for gestational age (AGA, N = 43), large for gestational age (LGA, N = 13) and small for gestational age (SGA, N = 12), according to birth weight. IgG subclasses (IgG1, IgG2, IgG3 and IgG4) were measured by the single radial immunodiffusion technique using monoclonal antibodies. IgG1 and IgG3 newborn subclass concentrations (10.17 and 0.57 g/l, respectively) increased with increasing gestational age and reached maternal levels (IgG1 = 8.86; IgG3 = 0.67 g/l) during the 37th week of pregnancy. Low levels of these subclasses were found in premature newborns. IgG2 from newborns were always lower than maternal levels (P < 0.05). LGA and SGA newborns had equivalent levels of IgG1 and IgG2 compared with AGA. SGA newborns had higher levels of IgG3 and lower levels of IgG4 than LGA and AGA newborns.

Transfer of IgG subclasses across placenta in term and preterm newborns

In order to study placental transfer of IgG subclasses, paired blood samples were collected from mothers and umbilical cord of preterm (N = 69) and full-term (N = 68) newborns. The full-term group was further divided into 3 subgroups: appropriate for gestational age (AGA, N = 43), large for gestational age (LGA, N = 13) and small for gestational age (SGA, N = 12), according to birth weight. IgG subclasses (IgG1, IgG2, IgG3 and IgG4) were measured by the single radial immunodiffusion technique using monoclonal antibodies. IgG1 and IgG3 newborn subclass concentrations (10.17 and 0.57 g/l, respectively) increased with increasing gestational age and reached maternal levels (IgG1 = 8.86; IgG3 = 0.67 g/l) during the 37th week of pregnancy. Low levels of these subclasses were found in premature newborns. IgG2 from newborns were always lower than maternal levels (P<0.05). LGA and SGA newborns had equivalent levels of IgG1 and IgG2 compared with AGA. SGA newborns had higher levels of IgG3 and lower levels of IgG4 than LGA and AGA newborns.