Agrobacterium-mediated transformation of the CrDAT gene and selection of transgenic periwinkle lines with a high vincristine accumulation

Catharanthus roseus contains vincristine and vinblastine, which are outstanding drugs for cancer. In the biosynthetic pathways of terpenoid indole alkaloids (TIAs) in C. roseus, deacetylvindoline 4-O-acetyltransferase (DAT) is a key enzyme that catalyses the last reaction of vindoline biosynthesis to form vinblastine and vincristine. In this study, the CrDAT transgene was transferred into the periwinkle by Agrobacterium-mediated transformation and generated transgenic periwinkle lines with an increase in vincristine accumulation. The C. roseus DAT gene was introduced into C. roseus plants and it was confirmed that CrDAT was successfully transferred into the genome of periwinkle plants and efficiently translated to synthesise recombinant DAT protein. Four transgenic periwinkle lines in T1 generation, T1-1, T1-3, T1-6, and T1-7, expressed recombinant DAT protein with the total protein content in the range of 2.86 μg.mg^?1 to 5.12 μg.mg^?1. Moreover, the vincristine contents of four transgenic lines increased by 1.63?2.48-fold compared to non-transgenic plants, ranging from 6.91 µg.g^?1 (fresh weight) to 10.53 µg.g^?1 (fresh weight). The T1-1 line had the highest vincristine content. Hence, the overexpression of the recombinant DAT protein can improve the vincristine accumulation of transgenic C. roseus plants.

Abbreviation: CrDAT - Catharanthus roseus Deacetylvindoline-4-O-Acetyl Transferase; D4H - Deacetoxyvindoline 4-hydroxylase; ELISA - Enzyme-Linked Immunosorbent Assay Monoterpene indole alkaloid; T0, T1 - Generations of transgenic plants; TIAs - Terpenoid indole alkaloids; WT- The wild-type tobacco plants (non transgenic plant); 35S - Cauliflower mosaic virus 35S promoter     

Effects of elevated CO2 concentration on bulbil germination and early seedling growth in Chinese yam under different air temperatures

The present study investigated the effects of elevated carbon dioxide concentration ([CO₂]) and air temperature on the germination of seed bulbils and the seedling vigour of two Chinese yam lines. Plants were grown under two [CO₂] levels, ambient and elevated (ambient + 200 μmol mol^?1), and two mean air temperature regimes, 22.2 °C (ambient + 1.4 °C) and 25.6 °C (ambient + 5.2 °C). Elevated [CO₂] did not affect bulbil germination under both air temperature regimes. During the early growth stage, the dry weight (DW) of leaves, vines, shoots, roots, belowground parts (roots + tubers) and whole plants were higher under elevated [CO₂] than ambient [CO₂] for both lines under the low- and high-temperature regimes. The values of vigour indexes (index I = germination % × seedling length and index II = germination % × seedling DW) were also higher under elevated [CO₂] than ambient [CO₂] for both lines. These results indicated that Chinese yam seedlings respond positively to elevated [CO₂] during the early growth stage. The above:belowground DW ratios were lower under elevated [CO₂] than ambient [CO₂] in seedlings with very small new tubers for both yam lines, indicating that elevated [CO₂] strongly affected the root growth in the early growth stage. The DWs of post-treatment seed bulbils were higher in the elevated [CO₂] under both air temperature regimes. The results showed that Chinese yam used a smaller amount of the reserves in seed bulbils under elevated [CO₂] than under ambient [CO₂].     

MAGNETIC RESONANCE, ULTRASOUND AND HISTOPATHOLOGIC CORRELATION OF ACUTE AND HEALING EQUINE TENDON INJURIES

Ultrasonography and MRI have become valuable tools for imaging of tendon injuries. The current study examines the histopathologic basis for the imaging abnormalities. Five injured equine forelimbs and two normal contralateral limbs were studied with high resolution real time ultrasound and MRI. Histologic sections were made and correlated with the diagnostic images. All lesions were readily seen by both modalities. Lesions characterized by hemorrhage, edma, and cellular infiltration were sonolucent on ultrasound and bright on MRI images. MRI returned to normal as fibrogenesis ensued. Ultrasound images remained abnormal until fibrillar reallgnment occured with completion of the healing process. High resolution real time ultrasound and MRI both accurately reflect the tissue abnormalities in acute tendon injuries. The injured horse does appear to be an effective model for correlative imaging studies of tendon injuries.     

Use of 64Copper Measurements to Diagnose Canine Copper Toxicosis

Inherited canine copper toxicosis is a serious problem in Bedlington terriers and West Highland White terriers, and may also be a problem in other less-studied breeds. Affected dogs become ill at midlife with progressive and ultimately fatal liver disease. Treatments for removal of copper and prevention of copper accumulation are available, but are most effective if begun before the dog becomes ill. Until recently diagnosis has not been available until the dog is 1 year of age, and then only by an invasive liver biopsy with determination of liver copper concentration. The authors studied the use of 64copper for early diagnosis of canine copper toxicosis. Two procedures were evaluated. The first involved measuring the concentration of 64copper in blood 24 hours after oral administration of the radioisotope. At this time, 64copper was associated primarily with ceruloplasmin secreted into the blood by the liver. This procedure is useful in the diagnosis of the human counterpart, Wilson's disease. However, the authors found it to be nondiscriminatory between affected and unaffected dogs. In contrast, the second procedure, which involved measuring 64copper excreted in stool during 48 hours after an intravenous dose of radioisotope, yielded results that differentiated most affected and unaffected dogs.     

USE OF 123IODINE METAIODOBENZYLGUANIDINE SCINTIGRAPHY FOR THE DIAGNOSIS OF A PHEOCHROMOCYTOMA IN A DOG

A 13-year-old neutered female Yorkshire terrier presented with a history of progressive episodic weakness and disorientation of 4 months duration. Physical and neurologic examinations were normal at presentation. Abdominal ultrasound revealed a mass involving the right adrenal gland. Standard planar scintigraphy was performed at 4, 18, and 24 hours after intravenous injection 185 MBq (5mCi) of ¹²³I-labeled metaiodobenzylguanidine (¹²³I-MIBG). An area of focal intense uptake was identified in the area of the right adrenal gland. A pheochromocytoma was confirmed histologically after surgical excision.     

Evaluation of Transmittance and Reflectance Pulse Oximetry in a Canine Model of Hypotension and Desaturation

Ten, anesthetized dogs were instrumented with three pulse oximeter probes; two lingual transmittance probes and one rectal reflective probe. Arterial oxygen desaturation was produced by decreasing the inspired oxygen concentration. Hypotension was produced with an infusion of nitroprusside. Simultaneous pulse oximeter readings (SpO₂) were compared to co-oximeter measured arterial saturation (SaO₂) collected over a range of SaO₂ (50–100%) and mean arterial pressures (40–100mmHg). Each of the monitors and means of evaluating SpO₂ studied provided accurate SpO₂ measurements over a range of mean arterial pressure from 40–100mmHg. All of the monitors tested tended to overestimate the SaO₂ when the arterial saturation was less than 70%.     

Effects of Fermentation Conditions Using Lactobacillus plantarum on the Charantin,Stigmasterol Glucoside and β-sitosterol Glucoside Contents of Bitter Gourd (Momordica charantia L.) Juice

Plant Foods for Human Nutrition - Bitter gourd fruits contain high amounts of charantin, stigmasterol glucoside and β-sitosterol glucoside, which have been shown to provide health benefits for...     

Differences in Ion Accumulation and Salt Tolerance among Glycine Accessions

Salinity reduces crop yield by limiting water uptake and causing ion‐specific stress. Soybean [Glycine max (L.) Merr.] is sensitive to soil salinity. However, there is variability among soybean genotypes and wild relatives for salt tolerance, suggesting that genetic improvement may be possible. The objective of this study was to identify differences in salt tolerance based on ion accumulation in leaves, stems and roots among accessions of four Glycine species. Four NaCl treatments, 0, 50, 75 and 100 mm , were imposed on G. max, G. soja, G. tomentella and G. argyrea accessions with different levels of salinity tolerance. Tolerant genotypes had less leaf scorch and a greater capacity to prevent Na⁺ and Cl^? transport from soil solution to stems and leaves than sensitive genotypes. Magnitude of leaf injury per unit increase in leaf Na⁺ or Cl^? concentrations was lower in tolerant than in susceptible accessions. Also, plant injury was associated more with Na⁺ rather than with Cl^? concentration in leaves. Salt‐tolerant accessions had greater leaf chlorophyll‐meter readings than sensitive genotypes at all NaCl concentrations. Glycine argyrea and G. tomentella accessions possessed higher salt tolerance than G. soja and G. max genotypes.     

Profile of Steroid Receptors and Increased Aromatase Immunoexpression in Canine Inflammatory Mammary Cancer as a Potential Therapeutic Target

Canine inflammatory mammary cancer (IMC) has been proposed as a model for the study of human inflammatory breast cancer (IBC). The aims of this study were to compare the immunohistochemical expression of aromatase (Arom) and several hormone receptors [estrogen receptor α (ERα), estrogen receptor β (ERβ), progesterone receptor (PR) and androgen receptor (AR)], in 21 IMC cases vs 19 non‐IMC; and to study the possible effect of letrozole on canine IMC and human inflammatory breast cancer (IBC) in vitro using IPC‐366 and SUM‐149 cell lines. Significant elevations of the means of Arom Total Score (TS), ERβ TS and PR TS were found in the IMC group (p = 0.025, p = 0.038 and p = 0.037, respectively). Secondary IMC tumours expressed higher levels of Arom than primary IMC (p = 0.029). Non‐IMC PR‐ tumours contained higher levels of Arom than non‐IMC PR+ tumours (p = 0.007). After the addition of letrozole, the number of IMC and IBC cells dropped drastically. The overexpression of Arom found and the results obtained in vitro further support canine IMC as a model for the study of IBC and future approaches to the treatment of dogs with mammary cancer, and especially IMC, using Arom inhibitors.