Specific B-cell Epitope of Per a 1: A Major Allergen of American Cockroach (Periplaneta americana) and Anatomical Localization

PURPOSE: Cockroach (CR) is a common source of indoor allergens, and Per a 1 is a major American CR (Periplaneta americana) allergen; however, several attributes of this protein remain unknown. This study identifies a novel specific B cell epitope and anatomical locations of Per a 1.0105. METHODS: Recombinant Per a 1.0105 (rPer a 1.0105) was used as BALB/c mouse immunogen for the production of monoclonal antibodies (MAb). The MAb specific B cell epitope was identified by determining phage mimotopic peptides and pair-wise alignment of the peptides with the rPer a 1.0105 amino acid sequence. Locations of the Per a 1.0105 in P. americana were investigated by immunohistochemical staining. RESULTS: The rPer a 1.0105 (~13 kDa) had 100%, 98% and > or =90% identity to Per a 1.0105, Per a 1.0101, and Cr-PII, respectively. The B-cell epitope of the Per a 1.0105 specific-MAb was located at residues99 QDLLLQLRDKGV110 contained in all 5 Per a 1.01 isoforms and Per a 1.02. The epitope was analogous to the Bla g 1.02 epitope; however, this B-cell epitope was not an IgE inducer. Per a 1.0105 was found in the midgut and intestinal content of American CR but not in the other organs. The amount of the Per a 1 was ~544 degrees Cg per gram of feces. CONCLUSIONS: The novel Per a 1 B-cell epitope described in this study is a useful target for allergen quantification in samples; however, the specific MAb can be used as an allergen detection reagent. The MAb based-affinity resin can be made for allergen purification, and the so-purified protein can serve as a standard and diagnostic allergen as well as a therapeutic vaccine component. The finding that the Per a 1 is contained in the midgut and feces is useful to increase yield and purity when preparing this allergen.

Specific B-cell Epitope of Per a 1: A Major Allergen of American Cockroach (Periplaneta americana) and Anatomical Localization

PURPOSE: Cockroach (CR) is a common source of indoor allergens, and Per a 1 is a major American CR (Periplaneta americana) allergen; however, several attributes of this protein remain unknown. This study identifies a novel specific B cell epitope and anatomical locations of Per a 1.0105. METHODS: Recombinant Per a 1.0105 (rPer a 1.0105) was used as BALB/c mouse immunogen for the production of monoclonal antibodies (MAb). The MAb specific B cell epitope was identified by determining phage mimotopic peptides and pair-wise alignment of the peptides with the rPer a 1.0105 amino acid sequence. Locations of the Per a 1.0105 in P. americana were investigated by immunohistochemical staining. RESULTS: The rPer a 1.0105 (~13 kDa) had 100%, 98% and > or =90% identity to Per a 1.0105, Per a 1.0101, and Cr-PII, respectively. The B-cell epitope of the Per a 1.0105 specific-MAb was located at residues99 QDLLLQLRDKGV110 contained in all 5 Per a 1.01 isoforms and Per a 1.02. The epitope was analogous to the Bla g 1.02 epitope; however, this B-cell epitope was not an IgE inducer. Per a 1.0105 was found in the midgut and intestinal content of American CR but not in the other organs. The amount of the Per a 1 was ~544 degrees Cg per gram of feces. CONCLUSIONS: The novel Per a 1 B-cell epitope described in this study is a useful target for allergen quantification in samples; however, the specific MAb can be used as an allergen detection reagent. The MAb based-affinity resin can be made for allergen purification, and the so-purified protein can serve as a standard and diagnostic allergen as well as a therapeutic vaccine component. The finding that the Per a 1 is contained in the midgut and feces is useful to increase yield and purity when preparing this allergen.

Effects of human contraceptive on reproduction and offspring in Chrysomya megacephala

OBJECTIVE@#To investigate the effect of human contraceptive (HC) as ability to suppress the reproductive success of blow fly, Chrysomya megacephala (Fabricius) (C. megacephala) and offspring under controlled laboratory conditions.@*METHODS@#Adult C. megacephala were fed with low (0.036 mg/mL) and high dose (0.072 mg/mL) HC (Microgest®, Thailand), containing levonorgestrel and ethinyl estradiol, in their drinking water for 7 days. Three experiments were set; experiment I with fed only in parental males, experiment II with fed only in parental females and experiment III with fed in both males and females. All experiments were then maintained for 3 generations after crossing and inbreeding.@*RESULTS@#A lower ovariole production and less fully mature ovarioles were evident in F1, F2 and F3 than control when parent males, females and both had been fed with high dose HC. Cellular changes during spermatogenesis in F1, F2 and F3 testes was confirmed using transmission electron microscopy (TEM), showing the low level of condensed chromatin, necrotic chromatin, irregularities and degenerated nuclear envelope in the nucleus. In the cytoplasm, mitochondrial swelling, rough endoplasmic reticulum swelling as well as vacuolated cytoplasm were noticed. As for the sperm per se, we found the degenerated nuclei and/or incomplete mitochondrial derivative, axoneme and vacuolated flagella. Regarding deformity in F1, F2 and F3 ovariole, ultrastructural alteration observed by scanning electron microscopy (SEM) included malformations involving fragile enveloping peritoneal sheath, cracked ovarioles, peel away chorion, crumbled eggshell and incomplete development; whereas TEM presented malformed and disorganized mass of cells, proteic yolk granules and vacuolated vesicles.@*CONCLUSIONS@#Administer of HC to adult C. megacephala caused ovariole reduction, less matured ovariole and affected cellular changes in testes and ovariole of offspring up to F3.

Peritrophic Membrane Formation in Adult Aedes aegypti after Blood Feeding

The peritrophic membrane (PM) is an acellular sheath that lines the digestive tract, separating the ingested food bolus from the midgut epithelium. It plays important roles in protecting the midgut epithelium from mechanical damage and insults from pathogens, and in facilitating food digestion. The purpose of this study was to examine the involvement of the PM in the midgut of Aedes aegypti mosquitoes after blood feeding. Ae. aegypti mosquitoes were fed with sucrose and blood, and collected at 0.5, 1, and 6 h post-oral feeding, respectively. Then, they were fixed in 2.5% glutaraldehyde in 0.1 M cacodylate buffer containing 5% sucrose pH 7.4 at 4°C, and prepared for study under a light microscope.PM was not produced when the mosquitoes received sucrose or fasted. Contrary to the blood feeding group, the PM clearly formed at 6 h post-feeding. It was concluded that PM construction was related to the ingestion of blood by the mosquitoes.