Determination of 1-deoxynojirimycin in mulberry leaves using hydrophilic interaction chromatography with evaporative light scattering detection

A simple and rapid method for determining 1-deoxynojirimycin (DNJ), a potent glucosidase inihibitor present in mulberry leaves (Morus alba and Morus bombysis), by high-performance liquid chromatography coupled to an evaporative light scattering detector (ELSD) has been developed. DNJ was separated from an extract of mulberry leaves on a TSKgel Amide-80 column, which is a representative column for hydrophilic interaction chromatography. During postcolumn detection, DNJ was detected by ELSD and concurrently identified by mass spectrometry. The detection limit was 100 ng. This method is sufficiently sensitive for determining DNJ in mulberry leaves and other related products.     

Use of genetic markers to identify the illegal trade of billfish in the second largest fishing warehouse of Latin America

1. Billfish are oceanic pelagic species that are often caught by tuna fleets and are of great interest for sport fishing. Two species of billfish have specific legislation prohibiting their marketing and export in Brazil.
2. DNA barcoding is a universal system of molecular identification based on a sequence of mitochondrial DNA cytochrome oxidase subunit I (COI), which serves as a diagnostic genomic marker in each species.
3. The barcode DNA technique was used to identify billfish marketed in the second largest fishing warehouse in Latin America, the CEAGESP (Companhia de Entrepostos e Armazéns Gerais de São Paulo), located in São Paulo, Brazil. Seventy‐nine samples of billfish were collected during three inspection visits carried out by Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis.
4. After DNA sequencing, 70 samples (88.60%) were identified to the species level; 21 (30.00%) were identified as Xiphias gladius, 43 (61.42%) as Istiophorus platypterus and six (8.57%) as Kajikia albida. The sale of this latter species is prohibited in Brazil and it is considered Vulnerable on the list of endangered species of the IUCN and in the official list of species of endangered Fauna – Fish and Aquatic Invertebrates.
5. Molecular analyses proved to be very efficient at uncovering irregularities in the identification of the white marlin (K. albida), which was traded illegally in CEAGESP, demonstrating the ineffectiveness of the current monitoring techniques used and emphasizing the need for the adoption of better public policies for the conservation of the species.

     

Characterization of <Emphasis Type="Italic">Inago1</Emphasis> and <Emphasis Type="Italic">Inago2</Emphasis> retrotransposons in <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

From the genome of a Japanese field isolate of the rice blast fungus, Magnaporthe oryzae, we newly identified Inago1 and Inago2 LTR retrotransposons. Both elements were found to be Ty3/gypsy-like elements whose copies were dispersed within the genome of Magnaporthe spp. isolates infecting rice and other monocot plants. Southern hybridization patterns of nine re-isolates derived from conidia of the strain Ina168 produced after a methyl viologen treatment were not changed, indicating that the insertion pattern of Inago elements is relatively stable.     

Molecular characterization of <Emphasis Type="Italic">Potato spindle tuber viroid</Emphasis> in dahlia

A new variant of Potato spindle tuber viroid (PSTVd) was detected for the first time from dahlia grown in Japan. The dahlia isolate of PSTVd formed a quasi-species and a major sequence variant consisting of 361 nucleotides in length, including five substitutions, three insertions, and one deletion, when compared to the intermediate strain from potato. In bioassays with the new isolate, Rutgers tomato developed mild stunting and leaf curling.     

Does paternal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) affect the sex ratio of offspring?

In 1976, men who were exposed to the highest concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) after an explosion at a chemical plant near Seveso, Italy, produced more girls than boys. However, few studies have examined the possibility that the exposure of laboratory animals to TCDD, especially that of males, could lead to a lower male/female sex ratio. The aim of this study was to investigate whether direct paternal exposure to TCDD affects the sex ratio of offspring using a relatively large-scale experimental design. Male ICR mice (n=120) were randomly assigned to three, one of which served as a vehicle control, the other two were administered TCDD orally with an initial loading dose of 2 or 2,000 ng TCDD/kg, followed by a weekly maintenance dose of 0.4 (T2/0.4 group) or 400 (T2000/400 group) ng/kg prior to mating. The major organs of each mouse were weighed and histopathologically and immunohistologically investigated, and the sex ratio of offspring [males/(males + females) x 100] was calculated in each dam. There were no significant effects on organ weights, or on the structure of the testis and epididymis between the control and TCDD-exposed males, but TCDD administration produced a significantly lower proportion of male offspring from T2000/400-exposed sires despite no alteration in litter size (Control: 53.1 +/- 1.7; T2/0.4: 48.8 +/- 2.5; T2000/400: 46.2 +/- 2.1). In addition, we further divided the T2000/400 group into 3 subgroups based on the proportion of CYP1A1-immunoreactive areas in the liver; there was a significant correlation between sex ratio and CYP1A1 immunoreactivity. Thus, the present study confirms that direct paternal exposure to TCDD might be associated with an alteration in the sex ratio of offspring. Possible mechanisms through which TCDD might decrease the fertility potential of Y-bearing gametes before conception are discussed.     

Peripartum changes in plasma estrone sulfate and estradiol-17beta profiles associated with and without the retention of fetal membranes in holstein-friesian cattle

The aim of this study was to evaluate the changes in plasma concentrations of estrone sulfate (E(1)S) and estradiol-17beta (E(2)beta) during the peripartum period (from day 10 prepartum to day 1 postpartum) associated with and without retention of fetal membranes (RFM) in Holstein-Friesian cattle (n=42). Plasma samples were analyzed for E(1)S and E(2)beta by ELISA. All parturitions were spontaneous and normal. Of 38 cattle delivering singletons, 29 had no RFM (singleton-normal group) and nine had RFM for more than 12 h (singleton-RFM group). Four cows gave birth to twins, and each twin had its own fetal membrane (FM). Two twinning cows expelled both FMs normally within 12 h (twin-normal group). In the remaining 2 twinning cows (twin-RFM group), the FM was expelled normally for one twin (first), while the FM of the other (second) was retained. There were no significant differences in the E(1)S concentrations or their increments from the concentrations on the preceding day between the normal and RFM groups of singleton cows on any peripartum day. The mean plasma E(2)beta concentrations on each day from day 10 to day 3 prepartum were significantly lower (P<0.05) in the singleton-RFM group compared with the singleton-normal group; however, on days 2 and 1 prepartum, the increments in the E(2)beta concentrations from the concentrations on the preceding days were significantly higher (P<0.05) in the singleton-RFM group than in the singleton-normal group. Thus, the plasma E(1)S concentrations just before parturition may not be associated with RFM. In the cows with RFM, the lower plasma E(2)beta concentrations that were found prior to day 2 prepartum may have been associated with immature placentomes, and the rapid rise in plasma E(2) beta within 1 to 2 days prior to calving may have produced asynchrony of placental and/or fetal maturation in relation to calving, thus resulting in RFM.     

Evaluation of a sequential extraction procedure for the fractionation of thallium in soils and determination of the content by flame atomic absorption spectrometry

Abstract

The release of carbonyl sulfide (COS) from 7 unamended paddy soils was estimated under aerobic and waterlogged conditions. The addition of organic materials to the soils increased the amount of COS evolved under waterlogged conditions. To assess the sources of the COS released from the soils, the soils were incubated under aerobic and waterlogged conditions after treatment with 17 inorganic or organic S-containing compounds. It was concluded that the following compounds represented sources of the COS released from the paddy soils: cystine, cysteine, thiocyanate, lanthionine and djenkolic acid.     

Synthesis of Hydroxy-Sodalite (“Zeolite”) from Waste Coal Ash

Abstract

Nitrogen accumulation in the apical spikelet of the top primary branch (superior spikelet) and the second spikelet of the lowest secondary branch (inferior spikelet), of the ear on the main stem of rice plant (Oryza sativa L. var. Sasanishiki) was characterized during grain filling. In the superior spikelet, the accumulation of dry matter and nitrogen which started immediately after lowering, proceeded rapidly, and reached the maturation level at 20 d after heading. In the inferior spikelet, however, the amount of dry matter and nitrogen accumulation was minimal immediately after flowering. It increased when grain filling of the superior spikelet was almost completed. ¹⁵N-labeled ammonia was administered to the plants at different stages of ripening and the amount of incorporation in the spikelets was analyzed at harvest. The labeled nitrogen administered at the early stages of ripening was the main source of the labeled nitrogen incorporated in the superior spikelet. However, the labeled nitrogen incorporated in the inferior spikelet largely consisted of the labeled nitrogen administered at the late stages of ripening. When all the spikelets except for the five inferior spikelets were removed from the ear at various stages of ripening, the amount of dry matter and nitrogen accumulation increased immediately, irrespective of the stage of ripening.

These results indicate that dry matter and nitrogen partitioning among spikelets of the ear is highly regulated during ripening and their accumulation in the inferior spikelets at the early stage of ripening is suppressed by some unknown factor(s) which can be eliminated by the removal of the superior spikelets.     

Rapid and sensitive detection of “<Emphasis Type="Italic">Candidatus</Emphasis> Liberibacter asiaticus” by cycleave isothermal and chimeric primer-initiated amplification of nucleic acids

We developed a detection method for “Candidatus Liberibacter asiaticus”, causal agent of citrus huanglongbing, using isothermal and chimeric primer-initiated amplification of nucleic acids combined with cycling probe technology (Cycleave ICAN). With Cycleave ICAN, the reaction was done in one tube in 1 h without the need for electrophoresis, and false positives were not generated. In addition, Cycleave ICAN method was more sensitive than the conventional PCR method. Cycleave ICAN helps shorten the time for the large-scale detection needed to manage huanglongbing.