ske-T, a T-box gene expressed in the skeletogenic mesenchyme lineage of the sea urchin embryo.

T-box transcription factors regulate many developmental processes. Here we report the cloning and expression analysis of ske-T, a novel sea urchin T-box gene. The distribution of the maternal ske-T transcript is uniform in the egg and early embryonic stages while zygotic expression is restricted to the skeletogenic mesenchyme lineage.

Allergenic extracts from Metarhizium canisopliae: obtainment and characterization.

Metarhizium anisopliae is used as a biopesticide for insects that damage agricultural plantations like sugar cane and forage plants. In a previous study the sensitization to this fungus of asthmatic patients coming from sugar cane areas was showed. The aims of this work were: to compare crude extracts obtained with Tris-HCl and Coca liquid from several growth phases of M. anisopliae concerning the total content of proteins and their electrophoretic analysis profile; to evaluate in vivo allergic sensitization in Balb/c mice and allergic patients from a sugar cane area, and to characterize the allergenic fractions in the sera of patients positive for the prick test by means of Western-blotting. The extract obtained with Coca liquid on the 16th day was the one that presented the greatest number of proteic fractions, including all those present in the other extracts. Twelve fractions were verified in this extract with approximate molecular weights from 94 to 14 kDa. The allergenicity of the extract obtained on the 16th day was proven by the production of IgE antibodies in Balb/c mice, with titres of 200. Prick tests carried out with the extract of the 16th day in 79 atopic individuals (from sugar cane area), 35 atopic individuals (from urban area) and 11 non- atopic individuals showed respective positivity of 29%, 9% and 0%. The allergenic characterization in vitro was performed by means of Western blotting, and the fractions that reacted with the positive individuals' sera were those of approximate molecular weights of 67 kDa (95%); 20 kDa (55%); 94 kDa (36%); 34 and 36 kDa (23%); 43 and 48 kDa (14%); 16 kDa (9%) and 54kDa (5%). It was concluded that the crude allergenic extract, obtained with Coca liquid from the 16th day growth of Metarhizium anisopliae, contains allergenic fractions and can be used in diagnostic screening tests.

Oral rehabilitation and psychosocial evaluation of a patient with bulimia nervosa following Teeth in a Day® immediate loading protocol.

PURPOSE: The purpose of this patient report is to accurately portray the clinical experience of treating an active bulimic patient by employing an immediately loaded dental implant protocol to provide the patient with an expedited esthetic and functional result. Implementation of a psychometric component statistically quantifies the psychological effects of the oral rehabilitation. MATERIALS AND METHODS: A 34-year-old woman presented with severe dental perimolysis (erosion) resulting from a 28-year history of bulimic behaviors. Clinical and radiographic evaluation of the existing dentition indicated advanced dental erosion and previous restoration attempts in a state of failure. The patient's existing teeth were extracted and treated with full maxillary and mandibular implant-supported prostheses, following the Teeth in a Day® (TIAD) immediate functional loading protocol. Psychometric assessment was conducted through administration of a 20-item questionnaire, as seen in previous literature. RESULTS: Radiographic and clinical analysis suggested all implant fixtures to be viable and accompanied by normal levels of alveolar bone 7 months into treatment. Questionnaire results at the new-patient presentation (T1) resulted in a composite score of 4.0 (SD = 2.9; P = 0.004), indicating severe body-image dysphoria. At 3 months postoperative (T2), the questionnaire score was 0.4 (SD = -1.7; P = 0.09). The final psychometric questionnaire at 7 months (T3) resulted in a score of 0.5 (SD = -1.5; P = 0.12). The patient was followed for a total of 7 years following implant placement. CONCLUSION: In this patient report, oral rehabilitation using the TIAD protocol provided an expedited and predictable alternative method for restoring functional abilities, esthetics, and self-esteem to a severely depressed bulimic patient. Psychometrically, at the postoperative (T2) and final testing period (T3), the patient's body image profile was no longer considered to be significantly different from established normative values.

House dust mites in the city of Lima, Peru.

Since mites are the most common house dust allergens, knowledge about the species most prevalent in a region is important for diagnostic and specific immunotherapy purposes. In order to establish the prevalence of house dust mites in different city districts, 100 house dust samples were collected from different parts of Lima. Lima is a city of tropical climate located along the coast of the Pacific Ocean. The relative air humidity is 80-90% and the various districts studied are located at altitudes ranging from 37-355 meters. The mite Blomia tropicalis was the organism most frequently detected, being present in 59% of the house dust samples. Dermatophagoides pteronyssinus occupied second place (15.9%), followed by Chortoglyphus arcuatus and Tyrophagus putrescentiae. These four mites, taken together, represented more than 90% of the mites detected. No specimen of the species Dermatophagoides farinae was detected. We conclude that B. tropicalis and D. pteronyssinus are the most common house dust mites in Lima. Considering the high prevalence of B. tropicalis in Lima and the fact that its cross-reactivity with antigens of the mites of the family Pyroglyphidae is minimal, we conclude that sensitization to this mite should be investigated separately in allergic patients living in Lima.

Library fungi at the University of São Paulo and their relationship with respiratory allergy.

In this study, we investigated the fungi most frequently found in the air and the books of 28 libraries at the University of São Paulo, Brazil. In a second phase, 314 librarians were questioned about the presence of asthmatic or rhinitic symptoms, and the relationship with the site of work. Forty-nine percent of them reported this type of symptoms and 80% related them to the place of work. In the third stage, librarians underwent intracutaneous tests against the 20 fungi most frequently isolated in libraries. Eighteen librarians presented positive tests, 12 of whom reported rhinitis and 6 of whom were asymptomatic. In 19.5% of symptomatic patients wheals > 15 mm were observed after 6 h, while 9% of the asymptomatic patients presented similar wheals. Fourteen librarians with positive tests underwent tests with each of the 20 fungi constituting the pool. The airborne fungi isolated in libraries are likely to be found anywhere in the city of São Paulo, but they are likely to be present in higher concentrations in libraries, subsequently producing respiratory allergies.

Bronchial challenge with aspirin lysine in the diagnosis of asthmatics with sensitization to aspirin and its inhibition by aerosolized furosemide.

We performed bronchial challenge with ASA lysine in 9 patients with a history of aspirin-induced asthma, 4 asthmatics with no history of hypersensitization to aspirin and 4 control subjects. The test consisted of successive inhalations of increasing concentrations of ASA lysine (11.25, 22.5, 45, 90, 180 and 360 mg/ml) and was interrupted when FEV1 showed a decrease of at least 20%. In order to determine the degree of bronchial hyperreactivity, we first carried out a bronchial challenge with histamine. All patients in the group sensitive to aspirin had a positive reaction to ASA lysine, while this was negative for patients in the two control groups. There was no significant correlation between PC20 to histamine and ASA lysine in any of the groups. On the other hand, 6 patients with sensitivity to ASA repeated the bronchial challenge with ASA lysine after previously inhaling furosemide, and in this second test, none of the 6 had a positive reaction. The variation of ASA lysine PC20 in both tests was positive for these patients (p < 0.001).

Characterization of allergenic fractions from Drechslera monoceras.

Drechslera monoceras, a fungus of the Deuteromycota phylum, is fairly frequent in Brazil, and is spread through the atmosphere. In previous studies done in the city of Sao Paulo, it was found that in relation to 42 other fungi extracts, the crude extract of this fungi demonstrated a more intense cutaneous reaction in patients with respiratory allergies. Biochemical, antigenic and allergenic evaluations were carried out at various growth stages of this fungus. Based on these facts, the purpose of this research was the fractionation and allergenic characterization of the allergenic extract of D. monoceras to be used in diagnosis and immunotherapy in patients with positive cutaneous reaction to this fungus. In the city of Sao Paulo, 13 of 248 patients with respiratory allergy (asthma and/or rhinitis) showed positive reaction following cutaneous tests (skin prick tests). The crude extract of D. monoceras was fractionated by SDS-PAGE. The visible fractions were then separated by electroelution to be inoculated into BALB/c mice to evaluate the production of IgE antibody. The IgE content was detected by passive cutaneous anaphylaxis test in Wistar rats, and two fractions of approximate molecular weights of 14.4 and 36 KDa reacted to the test. The in vitro allergenic characterization was carried out by Western blotting, and three fractions of approximate molecular weights of 14.4, 36 and 60 KDa were positive. It was concluded that the extract of D. monoceras has at least three allergenic determinants, which can be used for diagnosis and immunotherapy in patients with respiratory allergy to this fungi.

Candidin in immediate hypersensitivity. Comparison of two antigens.

One hundred outpatients in a Clinic of Allergy were submitted to intradermic tests with two types of candidins. The main focus of the research was the comparison of two antigens obtained from the same strains of Candida albicans: one a suspension of yeast cells and the other, a polysaccharide. The readings, taken 20 minutes after the intradermic injections, with positive results were considered as hypersensitivity of the immediate type. Positive results were obtained in 74% of the patients with the yeast cell antigen and in 73% with the polysaccharide antigen. This research mainly deals with the advantages that can be obtained by using the polysaccharide antigen in intradermic tests for evaluating hypersensitivity of the immediate type.

Acquired immunity to Africanized honeybee (Apis mellifera) venom in Brazilian beekeepers.

Seventy-eight Brazilian beekeepers who had been stung on average six times per month were studied. Sixty-eight beekeepers (87.1%) showed restricted local clinical reactions; nine individuals (11.5%) had extensive local reactions, and only one (1.2%) suffered anaphylactic shock. The humoral immunologic pattern of these individuals were studied by using immunoenzymatic methods to determine the serum titles of specific IgE and specific IgG4. Three groups of beekeepers presenting different humoral immunologic patterns were identified, in which the predominant pattern was the absence of specific IgE and high levels of specific IgG4 (38.4%). There was a positive correlation between the high levels of specific IgG4 and the number of bee stings. This correlation was not found in either specific or total IgE. The results of the present study suggest: i) the immunologic response to the number of exposures to Africanized honey-bee venom is not related to the number of exposures; and, ii) other nonhumoral and/or nonimmunologic factors may be involved in the reaction to the insect sting, which are responsible for both the clinical symptoms and protection.

Honeybee venom specific IgG subclass antibodies in Brazilian beekeepers and in patients allergic to bee stings.

Fifty-nine beekeepers who had been practicing apiculture for more than 2 years were selected in order to determine the distribution of bee venom specific IgG subclasses using ELISA. The assays were standardized into arbitrary units. For comparison, IgG subclasses were determined in eight individuals allergic to bee stings who did not receive specific treatment. No correlation was detected between beekeeping time and specific IgG1, IgG2 or IgG4 levels. There was a correlation between IgG2 levels and mean number of stings per month received by the beekeepers. Twenty-five percent of the beekeepers presented bee venom specific IgE class II or more in an ELISA assay. The IgG1 levels detected in beekeepers were similar to those detected in allergic individuals. IgG2 and IgG4 levels were significantly higher in beekeepers than in allergic individuals. IgG3 was not detected in any group studied. In conclusion, the maintenance of high levels of bee venom specific IgG2 and IgG4 represents a characteristic of beekeepers. These subclasses may be related to a modulatory effect of IgG on allergic reactions.

Extracts of Anacardium occidentale (cashew) pollen in patients with allergic bronchial asthma.

Allergic reactions to the pollen of trees is among the most prevalent allergic sensitivities. The cashew tree grows in abundance in the northeast region of the Brazil, mainly in Fortaleza city, in state of the Ceará. It flowers once a year between August and October. This is the first study conducted to establish the possible role of the cashew pollen extract in causing skin test reactivity in patients with allergic asthma. A stock solution of pollen extract was prepared with the standard weight/volume method for intradermal skin tests and for the protein content of the extract, estimated with the use of Folin phenol reagent and a spectrophotometer. Ten nonallergic volunteers and 80 subjects with allergic asthma, as documented by previous positive skin test reactions to various pollens, were studied. All of the 80 patients (100%) had positive test reactions (grade III and grade IV reactions). None of the control subjects (n = 10) had positive responses to the intradermal tests. This study provided us with knowledge of an additional pollen extract of the Anacardium occidentale, which could provoke skin test reactivities in asthmatic individuals from the northeastern area of Brazil. The results suggest a relationship between the period of flowering of the cashew tree and the increased number of allergic asthma cases.

Biochemical properties and study of antigenic cross-reactivity between Africanized honey bee and wasp venom.

Africanized honey bees and the wasp Polistes versicolor are common insects in Brazil; their venoms are composed of a complex mixture of components which present several biological activities. Stinging accidents are very frequent and are generally followed by important clinical reactions, and even deaths are not uncommon. In the present study, venom was extracted from Africanized honey bees and P. versicolor, and it was biochemically characterized and the antigenic cross-reactivity was investigated by Western blot analysis and specific IgE determination by ELISA in the sera of subjects allergic to each venom. The honey bee venom presented higher phospholipase A2 and hyaluronidase activities than P. versicolor venom, which in turn presented higher lipase, acid phosphatase and esterase activities. A high incidence of false-negatives was also observed during determinations of specific IgE for P. versicolor venom when the kits with venoms from wasps of temperate climates were used, suggesting that the diagnosis of allergy to neotropical wasp venom must take into consideration the clinical history and skin tests.

Candidin: comparison of two antigens for cutaneous delayed hypersensitivity testing.

A candidin, which is a suspension of killed yeast cells, is commonly used for intradermal tests of delayed hypersensitivity, to evaluate the immunological cellular competence of the patient, when the test is applied along with other similar tests. When working with a cellular antigen, the histopathology of positive skin tests reveals a cellular infiltrate which not only presents a characteristic hypersensitivity reaction but also a neutrophilic abscess in the central part. This research presents the results of a comparison between the yeast cell suspension and the polysaccharide antigens, both obtained from the same strains of Candida albicans. The results obtained by skin tests in one hundred individuals were 61.0% with the polysaccharide antigen and 69.0% with the yeast cell suspension antigen. Concordant results concerning the two antigens were observed in 82.0% of the individuals. The discussion section presents an assumption to explain the differences of positivity obtained with the two antigens. We conclude that the polysaccharide antigen can be utilized in the intradermal test of delayed hypersensitivity to Candida albicans.

[Immunology of tertiary pinta]. / Imunologia da pinta terciaria.

We've studied the immunological performed of twenty two natives Tikunas suffering from tertiary pinta. Among those, patients had been treated previously (two years earlier) which 2,400,000 IU of G benzathine penicillin, and twelve had no treatment. Both groups demonstrated an increment in the IgM synthesis (72.72%), IgG (50%), indicating the presence of strong antigenic stimuli. The great majority presenting a negative response revealed also a reduction in the cellular immune competence to at least two of the tests performed (92.3%), when were realized the PPD, DNCB and skin grafts tests.

[Hypersensitivity in patients with Hebra's prurigo caused by flea bite]. / Estudo da hipersensibilidade dos doentes com prurigo de Hebra à picada de pulga.

The authors present a study of hypersensitivity of patients with Hebra's prurigo (HP) to feleabites. Thirty six patients were studied. With the results obtained the following conclusions are held: 1. Among the probable responsible factors found in the history of patients with HP, the flea bite is the principal factor. 2. Almost all the patients with HP who were studied show hypersensitivity to flea bites. 3. With the flea bites on the patients with HP, one can observe the clinical and histopathology similarity of both the experimental and elementary lesions of the disease. 4. It was possible to show humoral antibodies in the serum of the patients with HP immunodiffusion technique to the 1:40 flea extract.

Cutaneous positivity in patients with respiratory allergies to 42 allergenic extracts of airborne fungi isolated in São Paulo, Brazil.

With the purpose of measuring the frequency of allergies of the respiratory tract to airborne fungi in São Paulo, Brazil, 201 patients with bronchial asthma and/or allergic rhinitis were submitted to intradermal testings with allergenic extracts of 42 fungus genera isolated from the air in São Paulo. The extracts of these fungi were prepared according to Coca's method and then standardized by the weight by the volume method. At first, the patients were submitted to intradermal testing with the total polyvalent (TP) extract, getting positive reaction in 70 of them (34.8%). The patients with positive reaction to the TP were tested with the 42 individual extracts and 74.3% of them reacted positively to one or more extracts. These results led us to the conclusion that the most frequent fungi in the air of São Paulo were not the ones which cause the highest number of positive intradermal reactions. Therefore, the convenience in employing as many allergens as possible in intradermal testings for diagnosis should be reinforced.

Systematic isolation of the genus Alternaria in the atmosphere of the "Grande Sao Paulo" area (Brazil).

The genus Alternaria has been isolated from the atmosphere of several countries and its spores are considered as having a strong allergenic power. The description of the genus isolation with a relative frequency was noted in the following countries: Mexico, the United States of America, Canada, Great Britain, Spain, South Africa, Denmark and Argentina. Many Brazilian authors have pointed out the rare incidence of this genus excluding Faria (5) who recorded in the city of Belo Horizonte, State of Minas Gerais, a frequency of 21.1%. Several authors have failed to detect Alternaria sp. spores in the atmosphere of the city of Sao Paulo. As we were studying in our laboratory the airborne fungi of a region called Greater Sao Paulo, where the collections were made in some boroughs of the main city and its neighborhood, we began to make findings of the Alternaria sp. This gave us the opportunity of studying its incidence in that area during a period of one year. We chose 11 areas of this region where we made weekly plate expositions performing 429 trials in all. The technique of collection was the usual one with the exposition of the Petri dishes containing Sabouraud Agar distributed 72 hours before. The collection was made from September 1973 to August 1974. The exposition period was of 5 minutes duration in the open air or in a protected place when it rained. The exposition height was 1 meter from the ground. The identification was made through the colony macroscopic morphology and through the Riddell technique. The Alternaria sp. was isolated in the mentioned area through all the months of the year with its highest frequency in August (35.8%) and its lowest in December 3.7%). The average relative frequency obtained was of 17%. The highest isolation frequency was attained in Winter, the lowest in Spring; in Summer and Autumn, it remained at a medium level. In relation to the studied areas, the Alternaria sp. was isolated in all of them with a minimum frequency of 8.3% in Campos Eliseos and with a maximum frequency of 28.9% in Osasco. The authors would like to emphasize this unusual finding because in the region studied the Alternaria sp. was previously considered absent.

Expression pattern of Brachyury in the embryo of the sea urchin Paracentrotus lividus.

Brachyury is a key transcription factor whose homologs have been identified in many animal species. Different Brachyury expression patterns have been observed amongst echinoderms. We have isolated PlBra, the Brachyury ortholog from the sea urchin Paracentrotus lividus and analyzed its expression during development. PlBra is first expressed at the end of cleavage in a ring of cells at the border between the presumptive endoderm and mesoderm territories. At later stages, PlBra is expressed around the blastopore and in the stomodaeum area as in most basal deuterostomes.

Study of humoral and cellular immunity in lobomycosis.

Fourteen patients with lobomycosis from the Amazon Basin were studied in order to assess their humoral and cellular immunological responses. The humoral response was normal, as shown by the normal levels of serum immunoglovulins. The cellular immunity was depressed for recent sensitizations as measured by skin contact test with DNCB (negative in 83.3% of 12 patients). The intradermal reactions to PPD and fungal extracts suggested that all patients were able to respond to at least 2 of the antigens used. It was also observed that patients reacting to PPD did not become sensitized to DNCB. Conversely, some Mantoux negative patients had positive reactions to DNCB. Both tests were negative in 3 patients.