Comparative analyses of the venom components in the salivary gland transcriptomes and saliva proteomes of some heteropteran insects.

Salivary gland-specific transcriptomes of nine heteropteran insects with distinct feeding strategies (predaceous, hematophagous, and phytophagous) were analyzed and annotated to compare and identify the venom components as well as their expression profiles. The transcriptional abundance of venom genes was verified via quantitative real-time PCR. Hierarchical clustering of 30 representative differentially expressed venom genes from the nine heteropteran species revealed unique groups of salivary gland-specific genes depending on their feeding strategy. The commonly transcribed genes included a paralytic neurotoxin (arginine kinase), digestive enzymes (cathepsin and serine protease), an anti-inflammatory protein (cystatin), hexamerin, and an odorant binding protein. Both predaceous and hematophagous (bed bug) heteropteran species showed relatively higher transcription levels of genes encoding proteins involved in proteolysis and cytolysis, whereas phytophagous heteropterans exhibited little or no expression of these genes, but had a high expression of vitellogenin, a multifunctional allergen. Saliva proteomes from four representative species were also analyzed. All venom proteins identified via saliva proteome analysis were annotated using salivary gland transcriptome data. The proteomic expression profiles of venom proteins were in good agreement with the salivary gland-specific transcriptomic profiles. Our results indicate that profiling of the salivary gland transcriptome provides important information on the composition and evolutionary features of venoms depending on their feeding strategy.

Development and validation of the Yin deficiency scale.

OBJECTIVES: Yin Deficiency (YD) is a pathophysiologic pattern that manifests with emaciation, dry mouth, tidal fever, Five Center Heat, night sweats, and malar flushing. The purpose of the present study was to develop and validate a new YD metric, the Yin Deficiency Scale (YDS), to define an optimum cutoff point for the YDS, and to examine the relationships between YD and Cold-Heat patterns. METHODS: A total of 169 outpatients and 83 college students were asked to complete the YDS. The college students were also asked to complete the Yin Deficiency Questionnaire (YDQ) and Cold-Heat Questionnaire (CHQ). Twelve clinicians determined whether or not the 169 outpatients had YD. YDS and YDQ data were used to estimate the internal consistency, construct validity, and concurrent validity of the YDS. CHQ data were used to examine the relationships between YDS and CHQ subscale scores. Total YDS scores and the clinicians' diagnoses of the outpatients were used to define an optimum cutoff score for the YDS. RESULTS: The 27-item YDS had satisfactory internal consistency (α=0.885) and concurrent validity, with correlations between the YDS and YDQ subscale scores. A total of eight factors were extracted in the test of construct validity, using principal component analysis, and the overall variance explained by all factors was 63.1%. Among the eight factors, cough, fever, and skin-related factors were associated only with Heat pattern, whereas pain, urine, and fatigue-related factors were associated with both Cold and Heat patterns. In the receiver operating characteristic curve analysis, the sensitivity, specificity, and area under the curve of the YDS were 73.7%, 84.8%, and 0.875, respectively. The optimum cutoff score was defined as 10 points. CONCLUSIONS: The results of this study suggest that the YDS is a reliable and valid instrument for evaluating YD.

The multimerization of hantavirus nucleocapsid protein depends on type-specific epitopes.

Multimerization of the Hantaan virus nucleocapsid protein (NP) in Hantaan virus-infected Vero E6 cells was observed in a competitive enzyme-linked immunosorbent assay (ELISA). Recombinant and truncated NPs of Hantaan, Seoul, and Dobrava viruses lacking the N-terminal 49 amino acids were also detected as multimers. Although truncated NPs of Hantaan virus lacking the N-terminal 154 amino acids existed as a monomer, those of Seoul and Dobrava formed multimers. The multimerized truncated NP antigens of Seoul and Dobrava viruses could detect serotype-specific antibodies, whereas the monomeric truncated NP antigen of Hantaan virus lacking the N-terminal 154 amino acids could not, suggesting that a hantavirus serotype-specific epitope on the NP results in multimerization. The NP-NP interaction was also detected by using a yeast two-hybrid assay. Two regions, amino acids 100 to 125 (region 1) and amino acids 404 to 429 (region 2), were essential for the NP-NP interaction in yeast. The NP of Seoul virus in which the tryptophan at amino acid number 119 was replaced by alanine (W119A mutation) did not multimerize in the yeast two-hybrid assay, indicating that tryptophan 119 in region 1 is important for the NP-NP interaction in yeast. However, W119A mutants expressed in mammalian cells were detected as the multimer by using competitive ELISA. Similarly, the truncated NP of Seoul virus expressing amino acids 155 to 429 showed a homologous interaction in a competitive ELISA but not in the yeast two-hybrid assay, indicating that the C-terminal region is important for the multimerization detected by competitive ELISA. Combined, the results indicate that several steps and regions are involved in multimerization of hantavirus NP.