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Anti-Bermuda grass RAST binding is minimally inhibited by pollen extracts from ten other grasses.

This article addresses the question, Does sensitization with Bermuda grass pollen generate IgE antibodies that cross-react with other grasses? We found that none of ten non-Bermuda grass extracts attained 50% inhibition of RAST binding between sera from patients in Arizona and Bermuda-coupled disks whereas Giant Bermuda grass extract was completely cross-reactive with Common Bermuda grass.

Binding Sites, Antibody

Immunologic and clinical correlates of bronchial challenge responses to Bermuda grass pollen extracts.

We have studied 26 patients with asthma who had positive skin tests for Bermuda grass pollen antigen (BGP) with history, immunologic assays, and bronchial challenge with BGP. In these patients IgE anti-BGP RAST titers were higher in those with positive, 6.8 +/- 4.4% (mean +/- SEM) compared to those with negative bronchial challenges, 2.7 +/- 0.4% (p less than 0.02). Historical data did not predict bronchial challenge response. IgG4 anti-BGP RAST was not different in patients with positive and negative challenges but was markedly increased, 15.3 +/- 11.9%, in patients who had previously had BGP immunotherapy when compared to untreated patients, 3.6 +/- 5.1% (p less than 0.01). IgE anti-Bermuda grass appears to be the major contributor to susceptibility to BGP bronchial challenge. IgG4 anti-Bermuda grass is apparently unrelated to the sensitivity to challenge but appears to be a significant component of the immunologic response to immunotherapy.

Antibodies, Anti-Idiotypic

Serum amylase activity and calcium and magnesium concentrations in young cattle grazing fescue and Bermuda grass pastures.

The study reported here was part of a long-term investigation of the effects of genotype on growth, reproduction, and metabolism in cattle grazing common Bermuda grass and endophyte-infected fescue pastures. In June 1990, blood samples were collected from the tail vein of yearling heifers and steers (Angus [AA], Brahman [BB], and their reciprocal crosses [AB, BA], n = 97). Serum amylase activity was assayed enzymatically; serum Ca and Mg concentrations were determined by atomic absorption spectrophotometry. The effects of endophyte-infected fescue depended on genotype (P less than 0.001). In yearlings having at least 1 Angus parent (AA, AB, BA), grazing endophyte-infected fescue was associated with higher serum amylase activity than was grazing Bermuda grass. But serum amylase activities of BB yearlings consuming either forage were similar. Moreover, for either forage, substantial differences were related to genotype (P less than 0.007) and gender (P less than 0.05). Angus yearlings had higher serum amylase activity than did Brahman yearlings; AB and BA yearlings had intermediate values. Heifers had higher amylase activity than did steers. The relationship among serum values of amylase, Ca, and Mg depended on forage. Yearlings consuming endophyte-infected fescue and having at least 1 Angus parent had a moderate negative correlation between serum amylase activity and Ca concentration (r = -0.53; P less than 0.0005); that is, in calves of genotypes with increased amylase activity while consuming endophyte-infected fescue (AA, AB, BA), the higher the amylase activity, the lower the serum Ca concentration. However, in yearlings consuming Bermuda grass, serum amylase and Ca values were not correlated.(ABSTRACT TRUNCATED AT 250 WORDS)

Amylases

Genome Report: De novo genome assembly of the greater Bermuda land snail, Poecilozonites bermudensis (Mollusca: Gastropoda), confirms ancestral genome duplication.

Poecilozonites bermudensis, the greater Bermuda land snail, is a critically endangered species and one of only two extant members in its genus. These snails are one of Bermuda's few endemic animal clades and their rich fossil record was the basis for the punctuated equilibria model of speciation. Once thought extinct, recent conservation efforts have focused on the recovery of the species, yet no genomic information or other molecular sequences have been available to inform these initiatives. We present a high-quality, annotated genome for P. bermudensis generated using PacBio long read and Omni-C short read sequencing. The resulting assembly is approximately 1.36 Gb with a scaffold N50 of 44.t Mb and 31 chromosome-length scaffolds. Nearly 43 percent of the genome was identified as repeat content. This assembly will serve as a resource for the conservation and study of P. bermudensis, and its only close extant and also critically endangered relative, P. circumfirmatus. Additionally, this genome adds to the growing body of data needed for a more complete understanding of gastropod evolution and for evolutionary processes in general.

Annotation

Asthma and climatic conditions: experience from Bermuda, an isolated island community.

A retrospective study of patients attending the emergency department with acute asthma was performed in Bermuda. Climatic data (barometric pressure, rainfall, humidity, and wind strength and direction) were obtained and compared with frequency of exacerbations of asthma. Three factors--namely, relative humidity, average daily temperature, and northeasterly winds--were found to be related to worsening asthma. Owing to Bermuda's lack of pollution and aeroallergens it was thought that these weather parameters had a direct effect on the asthmatic population.

Asthma

The preparation and testing of the proposed International Reference (IRP) Bermuda grass (Cynodon dactylon)-pollen extract.

A lyophilized candidate extract of Bermuda-grass (Cynodon dactylon) pollen, intended for use as an International Reference Standard, was prepared by pooling four individual candidate extracts. In preliminary investigations, the four candidate extracts encompassed a variety of extraction methods. The collaborative testing program simultaneously analyzed the proposed reference and the individual extracts and included 11 laboratories performing RAST inhibition, histamine release, crossed immunoelectrophoresis, crossed radioimmunoelectrophoresis, isoelectric focusing, sodium dodecyl sulfate gel electrophoresis, and protein determinations with a variety of reagents and methods. The four candidate extracts and the pooled reference were found to be equivalent. The stability of this extract has also been studied. This International Reference Preparation of Bermuda grass-pollen extract should be useful for research and industry.

Allergens

Identification of Bermuda grass (Cynodon dactylon)--pollen allergens by electroblotting.

Bermuda grass-pollen proteins were electrophoretically separated on polyacrylamide gels and transferred to nitrocellulose where IgE-binding components were detected by reaction with individual patient's serum and 125I-labeled antihuman IgE. Seventeen pollen components (in the molecular weight (MW) range of 8000 to 94,000 daltons), separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, bound IgE antibodies from a panel of 44 sera from allergic patients. The spectrum of Bermuda grass-pollen IgE-binding components detected is greater in number and wider in MW range than has previously been described. A component of MW 34,000 daltons (fraction 9) bound IgE from 100% of atopic sera tested. This component also bound the greatest quantity of IgE. Electrophoresis and transfer under nondissociating conditions revealed a component of MW 100,000 daltons that also bound IgE antibodies in all 44 sera tested. This component may be an aggregated form of fraction 9. A comparison of the electroblotting results obtained under dissociating and nondissociating conditions suggests once again that allergenic proteins in crude extracts may aggregate or associate during in vitro studies. Electrophoretic transfer analysis with sodium dodecyl sulfate-polyacrylamide gel electrophoresis may thus be the method of choice for allergen separation and identification.

Allergens

An immunoelectrophoretic analysis of the allergens of Cynodon dactylon (Bermuda grass) pollen.

A crude extract of Cynodon dactylon (Bermuda grass) pollen was obtained by extraction, centrifugation, dialysis, and lyophilization. The corresponding polyspecific rabbit antibodies were obtained by immunization, bleeding, and purification and were used for a cross immunoelectrophoretic analysis of the crude extract. At least 52 antigens (Ags), of which 47 migrated toward the anode, and five, toward the cathode, were revealed. Crossed radioimmunoelectrophoretic analysis performed with sera from 32 patients allergic to Bermuda grass and a pool from 1000 normal individuals revealed specific IgE binding to 13 of these Ags. The approximate molecular weights (MWs) for 10 of these IgE-binding Ags were determined by a combination of gel filtration and immunochemical analysis. These Ags had apparent MWs greater than 14 kd. A combination of preparative sucrose gradient isoelectric focusing and immunochemical analysis was used to determine the approximate isoelectric point values of five of the IgE-binding Ags. Most precipitating Ags had isoelectric points between 4.5 and 5.5. Four of the Ags (Ag 24, MW, less than 65 kd; Ag 31, MW, 33 kd; Ag 33, MW, 20 kd; and Ag 34/35, MW, 32 kd) were classified as major allergens.

Adolescent

Identification of allergens and antigens of Bermuda grass (Cynodon dactylon) pollen by immunoblot analysis.

Allergens and antigens of Bermuda grass pollen fractionated by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes were identified using twenty-one sera of Bermuda grass pollen-allergic patients. The IgE- and IgG-binding pollen components transferred to nitrocellulose were detected by reaction with enzyme-labelled anti-human IgE and anti-human IgG, respectively. There was heterogeneity in both IgE- and IgG-binding patterns of the allergic sera tested. Fourteen pollen components, ranging in molecular weight from 16,000 to 88,000 daltons, bound to IgE antibodies. Only two of the fourteen allergens identified reacted with IgE antibodies of more than 50% of the twenty-one allergic sera. The pollen component with a molecular weight of 32,000 daltons showed by far the highest frequency of IgE binding, being recognized by sixteen (76%) of the twenty-one sera examined. Fifteen (71%) of the twenty-one sera tested had IgE antibodies that reacted with more than one of the fourteen allergenic components identified. Pollen components recognized by IgE antibodies also reacted with IgG antibodies, and there were components only recognized by IgG antibodies. Results obtained from this study should be useful both clinically and in research.

Allergens

Nasopharyngeal cancer in Bermuda.

Six cases of nasopharyngeal carcinoma (NPC) were diagnosed in residents of Bermuda in the 7-year period 1966-1972. While epidemiologic study revealed no history of common environmental exposures, two of the six cases occurred in adult sibs from a family in which a third adult sib developed NPC in 1963. These three cases were from a large, possibly in-bred Portuguese family with origins in the Azores Islands. The remaining four cases were all in black children ages 10-15; two of these were first cousins. These observations suggest the possibility that genetic influence may contribute to the etiology of NPC.

Adolescent

Motorcycle injuries in Bermuda.

Tourists who hire mopeds or motorcycles for personal use have a risk for accident. The incidence of tourist accidents in Bermuda was 1.57 accidents/1000 tourists evaluated over a 6-month period. The accident incidence increased as the tourist population got larger. Tourists of more than 40 years of age had the greatest increase in incidence for accident during this period. The risk of an injury requiring hospital stay was one patient for every 16,000 tourists visiting the island, and usually involved the fracture of an extremity. All accidents, major or minor, resulted in some form of skin injury.

Accidents, Traffic

A clinical environmental study of the aeroallergens of the islands of Bermuda.

The major allergen responsible for allergic respiratory disease in the islands of Bermuda has been shown to be derived from Dermatophagoides pteronyssinus. 73% of the atopic group included in the survey gave weal and erythema reactions to extracts of this mite, whereas only 30% reacted to mixed pollen extracts and 10% to mould extracts. D. pteronyssinus was isolated from all house dust samples and it represented nature, with only a modest seasonal influence, and pollen counts were low throughout the year.

Adolescent

Characterization of the major allergen of Cynodon dactylon (Bermuda grass) pollen, Cyn d I.

An allergen from Cynodon dactylon (Bermuda grass) pollen, Cyn d I, has been purified by a combination of concanavalin A-Sepharose affinity chromatography, and carboxymethyl-Sepharose chromatography. The allergen constitutes the major allergenic component of the pollen extract as observed by immunoelectrophoretic techniques, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, an IgE-inhibition experiment, and skin testing. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Cyn d I is presented as a dominant 32 kd band and a minor 29 kd band, both binding IgE. Both bands are identified by monospecific rabbit antibodies (Abs) raised against Cyn d I. These Abs only weakly precipitate allergens from other grass species, indicating that Cyn d I possesses some unique immunochemical properties. Two of four purified murine monoclonal Abs raised against Cyn d I also bind to both bands of Cyn d I, indicating that the bands represent isoallergens with slightly different immunochemical properties. All four monoclonal Abs cross-react with pollen components from other grass species, especially Poa pratensis and Dactylis glomerata. The NH2-terminal sequence corresponding to approximately 10% of the complete sequence was determined, and it revealed high homology to the corresponding sequence of the major allergen of Lolium perenne, Lol p I. From the amino acid composition determination and immunoelectrophoretic comparison, the amount of Cyn d I in the source whole-pollen extract was estimated to be 15% wt/wt.

Allergens

Isolation and partial characterization of Bermuda grass pollen allergen, BG-60a.

In an earlier study we showed that Bermuda grass (Cynodon dactylon) pollen contains at least 12 IgE-binding proteins that can be analysed by immunoblot technique. One of the active components (BG-60) proved to be a basic protein of glycoprotein nature. It contained about 28% carbohydrate as determined from the dry weight and consisted of four molecules. One of the components was purified from the pollen extract by a combination of ammonium sulphate precipitation, ion-exchange chromatography on carboxymethyl-TSK, gel filtration on Ultrogel AcA 44 and chromatofocusing. Its molecular weight was approximately 60 kD by SDS-PAGE and 34 kD by gel filtration chromatography. The isoelectric point of the antigen was about 9.7. The homogeneity of the antigen BG-60a was assessed by one single arc of immunoprecipitation both in immunodiffusion and crossed immunoelectrophoresis and by one single band after SDS-PAGE. Its allergenicity was demonstrated by direct intradermal skin test on allergic patients and by examining IgE-binding reactivity with allergic patients' serum.

Adolescent

A unique human IgE-binding epitope on the Bermuda grass pollen recognized by mouse lambda-type monoclonal antibodies.

A group of six mouse monoclonal antibodies (MoAbs) with the unusual lambda-type light chain were generated by fusion of NS-1 cells with splenic cells derived from BALB/c mice immunized with crude extracts of Bermuda grass pollen (BGP). Four of them were IgG1, one was IgG2b, and one was IgG3. Binding inhibition assay showed that they recognized the same (or very similar) epitope. Using sera from BGP-allergic patients, it was found that the specific binding between the IgE antibodies and the MoAb 26-11-fixed antigen could be blocked by MoAb 26-11 itself and another MoAb 9-13 in a dose-dependent manner. It appears that the epitope recognized by the lambda-type MoAbs is a human IgE-binding antigenic determinant. Further physico-chemical analyses showed that this epitope was stable under heat but sensitive to treatments of sodium periodate and proteinase K. Results from these studies indicate that this unique epitope which leads to the generation of lambda-type MoAbs is part of a glycoprotein.

Animals

Analysis of allergenic components of Bermuda grass pollen by monoclonal antibodies.

A panel of 16 monoclonal antibodies (MoAbs) directed against Bermuda grass (Cynodon dactylon) pollen (BGP) were generated for identification and purification of the major allergenic components of the eliciting antigen (Ag). Radioimmunoprecipitation (RIP) analysis revealed that there were at least eight antigenic components with molecular weights (MW) ranging from 12 kilodalton (12 kDa) to 200 kDa. Each of these components has distinct biochemical characteristics based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing (IEF). Among them, Cyn d Bd67K and Cyn d Bd58K were basic proteins, Cyn d Bd35K consisted of at least four isomeric components with isoelectric points ranging from 6.2 to 7.2. The other antigens (Cyn d Bd68K, 48K, 38K, Cyn d Bd200K, Cyn d Bd46K, Cyn d Bd25K and Cyn d Bd12K) were all acidic proteins. The IgE binding capacity of all these antigens was determined with sera from 11 BGP-allergics by using a modified radioallergosorbent test. All but one of the antigens (Cyn d Bd200K) were found to react with human IgE from sera of BGP-allergic patients. Among those human IgE-binding molecules, Cyn d Bd35K reacted with allergic sera most frequently (10 of 11), followed by Cyn d Bd58K (8 of 11) and Cyn d Bd46K (7 of 11) respectively. Our results suggest that Cyn d Bd35K, Cyn d Bd58K, and Cyn d Bd46K are major allergens of BGP, and the MoAbs we obtained should be valuable tools for further purification of these allergens.

Allergens

Technical note: stocking equivalents and stocking rate-gain relationships for steers and cow-calf pairs grazing oversown bermuda grass.

Grazing experiments may use steers or cow-calf pairs for measuring animal performance on pasture treatments, but the validity of extrapolation between these classes of cattle has not been verified. A grazing study was conducted in the spring and summer of both 1988 and 1989 to determine stocking equivalents and stocking rate-weight gain relationships for steers and cow-calf pairs grazing Coastal bermuda grass (Cynodon dactylon [L.] pers.) oversown with rye (Secale cereale L.) and ryegrass (Lolium multiflorum Lam.). Average daily gain and stocking rate (SR; 3.2, 4.2, 6.2, and 7.4 animals per hectare for steers and 1.7, 2.5, 3.7, and 4.9 pairs per hectare for cow-calf pairs) were both adjusted so that comparisons could be made on an equal BW basis. Disk meter height readings were used as measurements of forage accessibility. Disk meter height responses to SR did not differ (P greater than .10) between steer and cow-calf paddocks. There was a linear (P less than .001) decrease in ADG as SR increased, but this decline was steeper (P less than .001) for steers than for cows or suckling calves. Steers tended to be more productive than calves at low SR but less productive at high SR. Disk meter heights for the range of SR used in the study did not differ (P greater than .10) for steers and cow-calf pairs at equivalent BW per hectare. Our study suggests that live BW is a reasonable basis for determining forage requirements of steers and cow-calf pairs under grazing conditions, but extrapolation of production between classes of livestock will not be reliable.

Animal Feed

Effects of supplemental Bermuda grass hay or corn on intake, digestion and performance of cattle consuming endophyte-infected fescue.

Effects of supplemental Bermuda grass hay (BG) or ground corn on intake, digestion and performance of cattle consuming endophyte-infected fescue (I) were studied. In Exp. 1, a Latin square study, five growing Holstein steers (158.1 kg) consumed I ad libitum and were offered 0, .3, .6, .9 or 1.2% body weight (BW) of BG daily. Total dry matter (DM) intake rose linearly (P less than .05) with increasing BG, although intake was numerically similar with .6, .9 and 1.2% BW of BG. Digestibility was constant with diet (P greater than .10). Six growing Holstein steers used in Exp. 2, a Latin square with a 2 x 3 factorial arrangement of treatments, ingested I or noninfected (NI) fescue hay ad libitum with 0, .5 or 1.0% BW of ground corn. Total DM intake increased linearly as the level of corn rose (P less than .05). Total intake with I increased more with the first than with the second addition of corn, and the opposite occurred with NI (interaction between fescue infection and the quadratic effect of corn level, P less than .10). Organic matter digested (g/d) was greater for NI than for I and rose linearly with increasing corn ingestion (P less than .05). Ninety-six crossbred beef heifers and steers (184.2 kg avg initial live weight) were used in a 77-d fall grazing experiment (Exp. 3) with a 2 x 3 factorial treatment arrangement. Cattle grazed I or NI paddocks and were given no supplement or .34% BW of BG or .65% BW of ground corn on a daily basis (DM).(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed