Cloning and functional analysis of the chitinase gene promoter in peanut.

Chitinase is an important pathogenesis-related protein in plants, and it can accumulate when induced by salicylic acid (SA) or other elicitors. Here, we found that chitinase mRNA levels were 4.5-times greater when peanut seedlings were sprayed with 1.5 mM SA, as compared to water. The upstream promoter sequence of the chitinase gene was cloned by TAIL-PCR and the potential cis-regulatory elements in this promoter were predicted by the cis-element databases PLACE and plantCARE. Elements in the promoter related to SA induction and disease resistance response included AS-1, GT1-motif, GRWAAW, TGTCA, W-box, and WB-box. The full-length promoter (P) and a series of 5'-deleted promoters (P1-P5) were cloned and then substituted for the 35S promoter of pCAMBIA1301-xylA, which carries the xylose isomerase gene as the selectable marker and GUS as the reporter gene. Six plant expression vectors (pCAMBIA1301-xylA-P-pCAMBIA1301-xylA-P5) were obtained. The six expression vectors were then transferred into onion epidermal cells and peanut plants by Agrobacterium-mediated transformation. Both the full-length and deleted promoters resulted in GUS staining of the onion epidermis cells when induced by SA. In onion epidermis cells, GUS enzyme activity was greater after SA induction. In transgenic peanut plants, GUS mRNA levels were greater after SA induction. Consideration of the cis-regulatory elements predicted by PLACE and plantCARE suggested that AS-1, GRWAAW, and W-box are positive regulatory elements in P2 and P3 and that GT1-motif and TGTCA are negative regulatory elements between P and P2.

Expression changes in epithelial cell adhesion molecule during colorectal cancer tumorigenesis.

We investigated the relationship between the expression of epithelial cell adhesion molecule (EpCAM) and the occurrence and development of colon cancer. Fifty colon cancer tissues and adjacent normal tissues were collected, while 40 normal intestinal mucosa tissues were collected as the blank group. EpCAM expression was detected by immunohistochemistry and the patients were followed-up to evaluate the prognosis. The positive expression rate of EpCAM reached 93.7% in patients with colorectal carcinoma, which was significantly higher than that in the negative control group and blank group (14.8 and 12.7%, P < 0.05, respectively). There was no significant difference between the control group and blank group regarding EpCAM expression. No direct relationship was observed between EpCAM expression and patient age, gender, and other characteristics. EpCAM was overexpressed in colorectal cancer and had a high detection rate. EpCAM can be used as a diagnostic biomarker for clinical detection of colorectal cancer.

New roles of filopodia and podosomes in the differentiation and fusion process of osteoclasts.

The cytoskeleton mediates various cellular processes such as differentiation and fusion, including in the filopodia and podosomes. However, apart from cell migration and formation of the sealing zone, little is known regarding the changes and related regulatory mechanisms of the cytoskeleton and additional roles of the filopodia and podosomes during the differentiation and fusion of osteoclasts. The cytomorphology and cytoskeleton of osteoclasts in the differentiation process were evaluated using tartrate-resistant acid phosphatase staining and immunofluorescence staining. Moreover, the expression levels of Rho GTPases and enzymes related to osteoclast differentiation and bone resorption were detected by quantitative reverse transcription-polymerase chain reaction. We detected 3 types of filopodia in osteoclast precursors and only 1 type of filopodia in undifferentiated cells. Mature osteoclasts were completely devoid of filopodia. Interestingly, cell fusion was highly specific, and the fusion initially occurred to the filopodia. Confocal images revealed that F-actin and microtubules significantly differed among fused cells. These results suggest that filopodia and podosomes not only play important roles in cell migration and the formation of sealing zones but also in the pre-fusion selectivity of 2 cells and the movement direction of the cell nucleus and cytoplasm during the fusion process. In addition, cdc42v1, RhoU, and RhoF regulate the formation of 3 types of filopodia during various stages of differentiation, while Rac1, Rac2, and filament A may be associated with cell selectivity during the fusion process.

Postmortem evaluation of 435 cases of intracranial neoplasia in dogs and relationship of neoplasm with breed, age, and body weight.

BACKGROUND: Intracranial neoplasia of dogs is frequently encountered in veterinary medicine, but large-scale studies on prevalence are lacking. OBJECTIVES: To determine the prevalence of intracranial neoplasia in a large population of dogs examined postmortem and the relationship between breed, age, and weight with the presence of primary intracranial neoplasms. ANIMALS: All dogs that underwent postmortem examination from 1986 through 2010 (n = 9,574), including dogs with a histopathologic diagnosis of primary (n = 227) and secondary (n = 208) intracranial neoplasia. METHODS: Retrospective evaluation of medical records from 1986 through 2010. RESULTS: Overall prevalence of intracranial neoplasia in this study's population of dogs was 4.5%. A statistically significant higher prevalence of primary intracranial neoplasms was found in dogs with increasing age and body weights. Dogs ≥15 kg had an increased risk of meningioma (odds ratio 2.3) when compared to dogs <15 kg. The Boxer, Boston Terrier, Golden Retriever, French Bulldog, and Rat Terrier had a significantly increased risk of primary intracranial neoplasms while the Cocker Spaniel and Doberman Pinscher showed a significantly decreased risk of primary intracranial neoplasms. CONCLUSIONS AND CLINICAL IMPORTANCE: Intracranial neoplasia in dogs might be more common than previous estimates. The study suggests that primary intracranial neoplasia should be a strong differential in older and larger breed dogs presenting with signs of nontraumatic intracranial disease. Specific breeds have been identified with an increased risk, and others with a decreased risk of primary intracranial neoplasms. The results warrant future investigations into the role of age, size, genetics, and breed on the development of intracranial neoplasms.

High segregation distortion in maize B73 x teosinte crosses.

Two genetic linkage maps of cultivated maize inbred lines and teosinte species were constructed. One population comprised 81 F(2) individuals derived from a cross between maize inbred line B73 and Zea mays ssp parviglumis, while the second consisted of 63 backcross individuals from a cross of maize inbred line B73 with Z. mays ssp diploperennis. In the B73 x Z. mays ssp parviglumis F(2) population, 172 simple sequence repeat (SSR) markers were mapped to 10 chromosomes, which covered 2210.8 cM. In the B73 x Z. mays ssp diploperennis backcross population, 258 SSR markers were mapped to 10 chromosomes, covering 1357.7 cM. Comparison of the two maps revealed that the total map length of Z. mays ssp diploperennis covers 1357.7 cM, which is about 61.4% of that of Z. mays ssp parviglumis (2210.8 cM). Extensive segregation distortion regions were found on chromosomes 1, 2, 3, 5, 6, 7, and 10 in the B73 x Z. mays ssp parviglumis F(2) population and on chromosomes 1-5 and 8-10 in the B73 x Z. mays ssp parviglumis backcross population. Segregation distortion analysis confirmed that the segregation distortion ratio in the interspecific population B73 x Z. mays ssp diploperennis was higher than in B73 x Z. mays ssp parviglumis. We found that the recombination distances are highly variable in these genetic crosses between cultivated and wild species of maize.

Remaining small bowel length: association with catheter sepsis in patients receiving home total parenteral nutrition: evidence of bacterial translocation.

Patients with short bowel syndrome (SBS) receiving total parenteral nutrition (TPN) have a high incidence of catheter-related sepsis, one of its major complications. The aim of this study was to correlate the length of remaining small bowel (RSB) with septic episodes related to the central venous catheter in a group of patients with severe SBS with home TPN. The length of the RSB (<50 cm or > or = 50 cm) was related to the frequency of catheter sepsis, time until the first episode, and the agents responsible in eight SBS patients receiving home TPN. There were 13 episodes of catheter infection (0.88 per patient-year). The group with a shorter RSB length (five patients) presented 1.3 to 2.76 infections/year and 2 to 9 months until the first episode, compared to 0 to 0.75 infections/ year (p = 0.0357) and 11 to 65 months until the first episode (p = 0.0332) in the group with the longer RSB. In the first group, the agents isolated were Enterobacteriae (Enterobacter sp., Klebsiella sp., Pseudomonas sp., and Proteus sp.) in eight episodes and Candida sp. in one. In the latter sepsis was caused by Staphylococcus sp. in three episodes and Pseudomonas sp. in one. Therefore patients with remaining small bowel shorter than 50 cm have a higher frequency of catheter-related sepsis, particularly by enteric microorganisms. This might be an evidence of the occurrence of bacterial translocation and its role in the pathogenesis of catheter-related sepsis in patients with an extremely short RSB receiving home TPN.