Macrophage-specific RAM11 monoclonal antibody cross-reacts with basal cells of stratified squamous epithelia.

RAM11 is a mouse monoclonal anti-rabbit macrophage antibody recognizing connective tissue and vascular (atheromatous tissue) macrophages. This study demonstrates a cross-reaction of RAM11 with an unknown antigen in rabbit normal epithelial cells. Formalin-fixed, paraffin sections of the New Zealand White rabbit normal skin, oral mucosa, esophagus, small intestine and lung were immunostained with RAM11 antibody followed by goat anti-mouse Cy-3-conjugated antiglobulin. RAM11-positive immunofluorescence was observed in basal layer cells of stratified squamous epithelia (skin, oral mucosa, esophagus). No RAM11 immunostaining was found in any cells of simple (intestinal, bronchial) epithelia. These findings show that basal cells of stratified squamous keratinized and non-keratinized epithelia of the rabbit express an antigenic epitope which is common with that of macrophage antigen recognized by RAM11 monoclonal antibody

Development of children's hymenoptera venom allergy quality of life scale (CHVAQoLS)

BACKGROUND: Venom allergy is a rare but life-threatening disease and may have a considerable impact on the health-related quality of life (HRQoL) of patients, especially children. This paper presents development of the HRQoL scale for children and adolescents with Hymenoptera venom allergy (HVA). METHODS: The study sample consisted of 71 children, born between 1992 and 2000, who presented with a history of insect sting reaction when referred for consultation in the allergy center of Polish-American Children’s Hospital, Krakow, Poland, during the period from 2000 to 2010. The initial pool of 60 items - divided into 6 domains - was prepared. The items with intercorrelations higher than 0.7 were removed from each domain and then principal component analysis was conducted for each domain separately, to provide a one-dimensional subscale for each domain. Reliability of the subscales was assessed using Cronbach alpha coefficient in terms of Classical Test Theory and with rho coefficient in terms of Item Response Theory. The multidimensionality of the scale was tested using multi-trait scaling. RESULTS: Three to four items from each domain were subsequently selected to constitute six subscales. Rho coefficients for all the subscales reached 0.8, similar results were achieved with the Cronbach alpha coefficients. Multi-trait method showed that the majority of the items indicated stronger correlations with their own subscales than with other subscales, which proves that our constructed subscales measure different dimensions of HRQoL. CONCLUSIONS: The presented scale comprises high validity and reliability subscales measuring six dimensions of HRQoL related to Hymenoptera venom allergy in children and adolescents. Such information may be useful in everyday clinical practice

Expression of basal cell marker revealed by RAM11 antibody during epithelial regeneration in rabbits.

RAM11 is a mouse monoclonal anti-rabbit macrophage antibody recognizing connective tissue and vascular macrophages. Our previous report showed that RAM11 reacted with basal cells of stratified squamous epithelia of rabbit skin, oral mucosa and esophagus. The aim of the present study was to follow the appearance of RAM11 immunoreactivity in basal cells of regenerating oral epithelium in rabbits. No RAM11 immunostaining was observed in the regenerating epithelium examined on days 1 and 3 of wound healing. A weak immunofluorescence first appeared on day 7 in single basal cells and 32% of RAM11- positive basal cells were observed on day 14. These findings indicate that expression of the antigen recognized by RAM11 antibody is a transient event in the differentiation of oral keratinocytes which not always occurs during epithelial repair, although it is a constant feature of epithelial turnover in mature epithelium. Therefore this antigen can be regarded as basal cell marker only in mature stratified squamous epithelia

Triple immunofluorescence labeling of atherosclerotic plaque components in apoE/LDLR -/- mice.

This paper presents a simple and reliable method of triple immunofluorescence staining that allows simultaneous detection of various cell types present in atherosclerotic plaque of apolipoprotein E and LDL receptor-double knockout (apoE/LDLR -/-) mice. We used combined direct and indirect procedures applying commercially available primary antibodies raised in different species to detect smooth muscle cells (Cy3-conjugated mouse anti-smooth muscle actin, SMA), macrophages (rat anti-CD68) and T lymphocytes (rabbit anti-CD3). Fixation of the material in acetone and modified incubation protocol employing nonfat dry milk in preincubation and incubation media significantly increased the intensity of labeling and effectively quenched the background. Our method offers an efficient way to detect qualitative as well as quantitative changes of macrophages, T lymphocytes and smooth muscle cells in atherosclerotic plaque of apoE/LDLR -/- mice during atherosclerosis development or in response to pharmacological treatment

Congruence of the current practices in Hymenoptera venom allergic patients in Poland with EAACI guidelines

Introduction: Venom immunotherapy (VIT) practice is the definitive treatment for patients with potentially fatal allergic reactions to Hymenoptera stings. The aim is assesing compliance of VIT practice in Poland with the current European Academy of Allergy and Clinical Immunology (EAACI) guidance. Material and methods: A multicentre study was carried out using a structured questionnaire which was sent by post to all VIT practitioners in Poland. Some questionnaire items were altered, in comparison to original version by adding additional answer options or alowing multiple answer option. The response rate was 100%. The obtained results were compared with the published EAACI guidelines. Results: Twenty-six Polish centres took part in the survey. SSIgE and skin prick tests (SPT) are together used as the first line of investigation, whereas confirmatory intradermal tests (IDT) are applied in half of centres. Only a few centres measure baseline serum tryptase levels. The ultra-rush protocol is preferred. Antihistamine pre-medication is routinely practiced. A target dose equal to 100 µg is used in most centres. A 6-week interval between booster doses is the most frequent. Five years is considered as an optimal VIT duration. Before the VIT completion, SSIgE is evaluated in fifty percent of centres, whereas sting challenge is considered by half of responders. Conclusions: There are some differences between current practice in Poland and the EAACI recommendations, indicating areas requiring better compliance. Comparision between Poland and the United Kingdom revealed that health service organization and health care funding may play a major role in the provision of allergy services. This may affect the extent to which international guidance may be applied in individual countries. It is worth considering conducting the same survey in other European countrie

Identification of Secreted Exoproteome Fingerprints of Highly-Virulent and Non-Virulent Staphylococcus aureus Strains

Staphylococcus aureus is a commensal inhabitant of skin and mucous membranes in nose vestibule but also an important opportunistic pathogen of humans and livestock. The extracellular proteome as a whole constitutes its major virulence determinant; however, the involvement of particular proteins is still relatively poorly understood. In this study, we compared the extracellular proteomes of poultry-derived S. aureus strains exhibiting a virulent (VIR) and non-virulent (NVIR) phenotype in a chicken embryo experimental infection model with the aim to identify proteomic signatures associated with the particular phenotypes. Despite significant heterogeneity within the analyzed proteomes, we identified alpha-haemolysin and bifunctional autolysin as indicators of virulence, whereas glutamylendopeptidase production was characteristic for non-virulent strains.Staphopain C (StpC) was identified in both the VIR and NVIR proteomes and the latter fact contradicted previous findings suggesting its involvement in virulence. By supplementing NVIR, StpC-negative strains with StpC and comparing the virulence of parental and supplemented strains, we demonstrated that staphopain C alone does not affect staphylococcal virulence in a chicken embryo model

Theoretical validity and reliability of Vespide Quality of Life Questionnaire in Polish adolescents with Hymenoptera venom allergy

Measurement of health-related quality of life is mostly used with advanced measurement tools, such as scales. The use of a certain scale in different cultural settings or in age groups other than those it was designed for needs conducting the adaptation process and assessment of psychometric properties of the adapted scale. This paper presents the results of theoretical validity and reliability analysis of the Polish adaptation of the VQLQ scale for adolescents with Hymenoptera venom allergy. The study sample consisted of 78 adolescents aged 14-19 years, who were treated with venom immunotherapy in Polish allergological centers in 2008 year. Theoretical validity of the scale was analyzed with exploratory factor analysis using the principal components method. Reliability analysis was assessed in terms of internal consistency with Cronbach's α coefficient and by testing Kline's criterion. The results showed satisfactory validity of the scale: factor analysis revealed a 3-factor structure of the scale - extracted factors were described as anxiety, caution and discomfort. All the scale items contributed to unique factors, except for one item concerning limitation in summer due to allergy, which was identified as a separate dimension of health-related quality of life of Polish adolescents with Hymenoptera venom allergy. All the extracted subscales were characterized by values of α coefficient equal or higher than 0.8, what is usually considered as a high-level reliability coefficient. The adapted scale is a valid and reliable tool measuring health-related quality of life in Polish adolescents treated with venom-specific immunotherapy

Health-related quality of life in Polish adolescents with "Hymenoptera" venom allergy treated with venom immunotherapy

Introduction: Venom allergy, though rare, may seriously influence health-related quality of life (HRQoL). There is a paucity of research on HRQoL of adolescents and young adults with Hymenoptera venom allergy. The aim was to assess the level of HRQoL and to evaluate its independent predictors in Polish adolescents and young adults treated with venom immunotherapy. Material and methods: A multicenter cross-sectional study based on the Vespid Allergy Quality of Life Questionnaire (VQLQ) adapted for Polish adolescents was used. The study sample included 87 patients (14-21 years) studied at different stages of venom immunotherapy (VIT). Statistical analysis was done with multivariate linear regression. Results: Anxiety level was higher in patients with 4th grade of Mueller’s classification (anaphylactic shock) than in those with 3rd grade (B = 0.84, 95% CI = 0.07-1.61, p = 0.03). Caution increased along with an increase of anxiety of adolescents treated with VIT (B = 0.54, 95% CI = 0.39-0.68, p < 0.01). Level of limitations increased with increasing caution of adolescents (B = 0.63, 95% CI = 0.35-0.91, p < 0.01). Discomfort increased along with a rise of caution of patients (B = 0.38, 95% CI = 0.22-0.55, p < 0.01). Similarly, it increased with an increase of their feeling of limitations (B = 0.37, 95% CI = 0.23-0.51, p < 0.01). The level of discomfort in adolescents treated with VIT was lower in those who were treated with conventional protocol in comparison to those treated with rush or ultrarush ones (B = –0.47, 95% CI = –0.90 - –0.03, p = 0.04). Conclusions: Severity of anaphylactic reaction is an independent determinant of anxiety level in adolescents treated with VIT. The VIT protocol affects HRQoL of treated patients