A More Complete Story of the Genetic Bases of Resistance to the Rice Hoja Blanca Virus

Rice hoja blanca is one of the most serious diseases in rice growing areas in tropical Americas. Its causal agent is the Rice hoja blanca virus (RHBV), transmitted by the planthopper Tagosodes orizicolus Müir. Genetic resistance is the most effective and environment-friendly way of controlling the disease. So far, only one major quantitative trait locus (QTL) of Oryza sativa ssp. japonica origin, qHBV4.1, that alters incidence of the virus symptoms in two Colombian cultivars has been reported. This resistance has already started to be broken, stressing the urgent need for diversifying the resistance sources. In the present study we performed a search for new QTLs of O. sativa indica origin associated with RHBV resistance. We used four F2:3 segregating populations derived from indica resistant varieties crossed with a highly susceptible japonica pivot parent. Beside the standard method for measuring disease incidence, we developed a new method based on computer-assisted image processing to determine the affected leaf area (ALA) as a measure of symptoms severity. Based on the disease severity and incidence scores in the F3 families under greenhouse conditions, and SNP genotyping of the F2 individuals, we identified four new indica QTLs for RHBV resistance on rice chromosomes 4, 6 and 11, namely qHBV4.2WAS208, qHBV6.1PTB25, qHBV11.1 and qHBV11.2. We also confirmed the wide-range action of qHBV4.1. Among the five QTLs, qHBV4.1 and qHBV11.1 had the largest effects on incidence and severity, respectively. These results provide a more complete understanding of the genetic bases of RHBV resistance in the cultivated rice gene pool, and can be used to develop marker-aided breeding strategies to improve RHBV resistance. The power of joint- and meta- analyses allowed precise mapping and candidate genes identification, providing the basis for positional cloning of the two major QTLs qHBV4.1 and qHBV11.1.

Quantification by computerized morphometry of tissue levels of sulfomucins and sialomucins in diversion colitis in rats

PURPOSE: To quantify the intensity of sulfomucin and sialomucin expression in the colon mucosa, by means of computer-assisted image processing, comparing segments with and without fecal stream and correlating with the duration of fecal transit exclusion. METHODS: Forty-five Wistar rats were subjected to diversion of the fecal stream in the left colon by means of constructing a proximal colostomy and distal mucosal fistula. They were distributed randomly into three experimental groups of 15 animals, of which 10 were subjected to colon diversion (experimental subgroup) and five were only subjected to laparotomy, without colon diversion (control subgroup). The three experimental groups were formed according to the sacrifice date, which was to be performed six weeks after the surgical procedure (Group A), 12 weeks (Group B) and 18 weeks (Group C). The sulfomucin and sialomucin expression in the colon mucosa was evaluated using the histochemical technique of high iron diamine-alcian blue (HID-AB). The tissue expression was quantified for each animal, in the segments with and without fecal stream, at a location where there were four complete contiguous crypts in two random fields, with the aid of the computer-assisted image analysis software. The final value was taken to be the mean reading from the two fields selected, in the segments with and without fecal stream. To compare the expressions of the two mucin subtypes in the segments with and without fecal stream, the paired Student t test was used. To analyze variance according to duration of exclusion, ANOVA with the Newman-Keuls post-test was used, setting the significance level at 5% (p<0.05). RESULTS: There were significant reductions in tissue sulfomucin and sialomucin content in the colon without fecal stream, independent of the duration of exclusion considered. There was increased tissue sulfomucin content and decreased tissue sialomucin in the segments without fecal stream, with increasing duration of exclusion. CONCLUSIONS: Diversion of the fecal transit decreased the tissue sulfomucin and sialomucin content in the segments without fecal stream. Notwithstanding the reduction in the levels of both subtypes of acid mucin in the segments without fecal stream, there was increased tissue sulfomucin content and decreased tissue sialomucin with increasing duration of intestinal diversion.

Apoptotic effects of inositol hexaphosphate on biomarker Itpr3 in induced colon rat carcinogenesis

PURPOSE: To study the effect of the modulation of inositol hexaphosphate (IP6) in the biological immunohistochemistry expression of cellular signaling marker apoptosis, in model of carcinogenesis of colon induced by azoxymethane (AOM). METHODS: Wistar rats (N=112) distributed in 4 groups (n=28): Control; B, AOM (5 mg kg-1, 2x, to break week 3); C, IP6 (in water 1%, six weeks); D, IP6+AOM. Weekly euthanasia (n=7), from week three. Immunohistochemistry of ascendant colon with biological marker inositol 1,4,5 triphosphate receptor type III (Itpr3). Quantification of the immune-expression with use of computer-assisted image processing. Analysis statistics of the means between groups, weeks in groups, groups in weeks, and established significance when p<0.05. RESULTS: One proved significant difference between groups in the expression of Itpr3, p<0.0001; with Itpr3 reduction of BxD group, p<0.001. CONCLUSION: Inositol hexaphosphate promotes modulation of biological markers with reduction of Itpr3 in carcinogenesis of colon.

Reliability of the WIDERYST Susceptibility Testing System for Detection of In Vitro Antifungal Resistance in Yeasts

ABSTRACT This study evaluated the WIDERYST system, a commercially available computer-assisted image-processing device for the antifungal susceptibility testing of yeasts. A collection of 90 clinical isolates selected to represent ranges of susceptibilities in vitro as broad as possible was tested. An evaluation compared the results obtained by the new system with those achieved by both the Clinical and Laboratory Standards Institute (CLSI) microdilution reference procedure and the antifungal susceptibility standard of the European Committee for Antimicrobial Susceptibility Testing (EUCAST). Overall, the agreement and the correlation index between results obtained by the EUCAST method and the WIDERYST system were 89% and 0.84 (P < 0.01), respectively, and agreement and correlation index between data obtained by the CLSI procedure and the WIDERYST system were 90% and 0.86 (P < 0.01), respectively. The system was able to detect amphotericin B-resistant isolates. All Candida sp. isolates with resistance in vitro to azole agents were detected as well. The system misclassified some isolates belonging to the slowly growing genera Dipodascus and Pichia. A total of 2.7% very major errors were detected for fluconazole. The WIDERYST system is an alternative to reference procedures for antifungal susceptibility testing of clinical isolates of yeasts, particularly for Candida and Cryptococcus species.