Role of source-to-sink transport of methionine in establishing seed protein quantity and quality in legumes.

Grain legumes such as pea (Pisum sativum L.) are highly valued as a staple source of protein for human and animal nutrition. However, their seeds often contain limited amounts of high-quality, sulfur (S) rich proteins, caused by a shortage of the S-amino acids cysteine and methionine. It was hypothesized that legume seed quality is directly linked to the amount of organic S transported from leaves to seeds, and imported into the growing embryo. We expressed a high-affinity yeast (Saccharomyces cerevisiae) methionine/cysteine transporter (Methionine UPtake 1) in both the pea leaf phloem and seed cotyledons and found source-to-sink transport of methionine but not cysteine increased. Changes in methionine phloem loading triggered improvements in S uptake and assimilation and long-distance transport of the S compounds, S-methylmethionine and glutathione. In addition, nitrogen and carbon assimilation and source-to-sink allocation were upregulated, together resulting in increased plant biomass and seed yield. Further, methionine and amino acid delivery to individual seeds and uptake by the cotyledons improved, leading to increased accumulation of storage proteins by up to 23%, due to both higher levels of S-poor and, most importantly, S-rich proteins. Sulfate delivery to the embryo and S assimilation in the cotyledons were also upregulated, further contributing to the improved S-rich storage protein pools and seed quality. Overall, this work demonstrates that methionine transporter function in source and sink tissues presents a bottleneck in S allocation to seeds and that its targeted manipulation is essential for overcoming limitations in the accumulation of high-quality seed storage proteins.

Three­Dimensional Ultrasonography in Preoperative and Postoperative Volume Assessment of the Undescended Testicle.

BACKGROUND Ultrasound (US) is the preferred imaging method for cryptorchidism, but most guidelines indicate that its value is questionable. The aim of this study was to evaluate the clinical value of ultrasonic mobility and testicular atrophy index (TAI) based on three­dimensional US (3DUS) in preoperative and postoperative assessment of the undescended testis. MATERIAL AND METHODS Data from 158 children with unilateral extraperitoneal cryptorchidism were collected and their diagnoses were surgically confirmed. They were divided into different age groups and into 2 ultrasonic mobility groups: the mobile group (MG) and the restricted group (RG). Differences in sonographic characteristics between different groups were compared. Three-dimensional ultrasound performed with virtual organ computer-aided analysis (VOCAL) was used to determined preoperative and postoperative TAI and the reliability of TAI was analyzed. RESULTS Measurement of testicular volume with the VOCAL method was significantly more reliable than that done with the two-dimensional Lambert method. In all age groups, preoperative testicular volumes were smaller than that in the contralateral scrotal testis and postoperatively, they increased steadily. Both preoperative and postoperative TAI were higher in the RG than in the MG. In the MG, postoperative TAI decreased significantly in all age groups. In the RG, in contrast, effective volume growth was only achieved in patients who had undergone surgery before they reached age 1 year. CONCLUSIONS TAI values determined with 3DUS using the VOCAL technique objectively reflect recovery of testicular volume following surgery for undescended testicle. Ultrasonic mobility evaluation is beneficial for clinical management of the condition.

Correlation of adrenomedullin with the erythropoietin receptor and microvessel density in hepatocellular carcinoma.

INTRODUCTION: Uncontrolled angiogenesis plays an essential role in the occurrence, metastasis and malignant progression of hepatocellular carcinoma (HCC). This study aimed to investigate the expression of adrenomedullin (ADM) in human HCC and its correlation with the expression of erythropoietin receptor (EPOR), microvessel density (MVD) and the tumor pathological characteristics. MATERIAL AND METHODS: Fresh tumor tissues were obtained from 30 HCC patients after hepatectomy. Ten cirrhotic and 10 normal liver tissues were included as controls. Expression of ADM and EPOR was determined by real-time PCR. The MVD was determined by counting the number of microvessels. RESULTS: The MVD and the mRNA levels of ADM and EPOR in cancer tissues were significantly higher than those in the non-cancer tissues (p < 0.05). Expression of ADM was significantly correlated with the MVD and EPOR (r = 0.68 and 0.74, p < 0.01). Adrenomedullin and EPOR mRNA levels in HCC tissues were correlated with capsule invasion, pathological differentiation and tumor metastasis (p < 0.05). CONCLUSIONS: Our findings suggest that ADM and EPOR may serve as new regulatory factors involved in angiogenesis of HCC and represent novel targets for the treatment of HCC.

Elevated expression of MDR1 associated with Line-1 hypomethylation in esophageal squamous cell carcinoma.

BACKGROUND: The aim is to discuss the relationship of Line-1 methylation and the MDR1 expression in esophageal squamous cell carcinoma (ESCC). METHODS: We analyzed the methylation level of Line-1 by quantitative real-time MSP, and the expression of MDR1 by real-time RT-PCR in 310 ESCC and corresponding non-tumor tissues. RESULTS: We found that the methylation index (MI) of Line-1 decreased from 0.90 in non-tumor tissues toward 0.78 in ESCC. The cumulative survival was significantly shorter in ESCC patients with MI ≤ 0.78 (34 months) than that in patients with MI > 0.78 (43 months). There was a statistical difference between MI ≤ 0.78 and MI > 0.78 cases with these clinicopathologic parameters (age, AJCC stage, differentiation; P = 0.010, P < 0.0001, P = 0.015, respectively). These results implied that Line-1 hypomethylation could be more in ESCC patients with older, advanced tumor and poor differentiation group. Meanwhile, ESCC with demethylation of Line-1 were shown elevated MDR1 expression in tumor (Mean-∆∆Ct = 0.21), but ESCC with hypermethylation of Line-1 were considered to be decreased MDR1 expression in tumor (Mean-∆∆Ct = -0.86). CONCLUSIONS: Line-1 hypomethylation could be as a biomarker of poor prognosis in ESCC patients. MDR1 gene could be activated via epigenetic mechanisms with demethylation of Line-1 in ESCC, and enhance tumor progression.

Iron-catalyzed cascade carbochloromethylation of activated alkenes: highly efficient access to chloro-containing oxindoles.

An iron-catalyzed carbodi- and trichloromethylation of activated alkenes with readily available dichloro- and tetrachloromethane has been developed. A diaryliodonium salt is used as an efficient oxidant in this transformation. This reaction tolerates a variety of functional groups and allows for a highly efficient synthesis of various chloro-containing oxindoles.

[Immune tolerance induced by bone marrow cell transplantation combined with use of cyclophosphamide in diabetic rats with pancreatic transplantation].

OBJECTIVE: To study the immune tolerance induced by bone marrow cell transplantation combined with short-term use of cyclophosphamide after pancreatic transplantation in diabetic rats. METHODS: Type I diabetes mellitus was induced in BN rats with streptozotocin (STZ) intraperitoneal injection at a single dose of 45 mg/kg. Pancreatic transplantations were performed with the SD rats as donors and the diabetic BN rats as recipients. Twenty BN rats with type I diabetes mellitus were randomly divided into four groups. The BN rats in Group I received pancreas transplantations only. The BN rats in Group II received intraperitoneal injection of 150 mg/kg cyclophosphamide on the first day after pancreas transplantations. The BN rats in Group 11 received injection of 2.0 x 10(8) donors' bone marrow cells via the portal vein during the pancreas transplantations. The BN rats in Group IV received injection 2.0 x 10(8) donors' bone marrow cells via the portal vein during the pancreas transplantations and an intraperitoneal injection of 150 mg/kg cyclophosphamide on the first day after pancreas transplantations. The blood glucose of the rats was measured after transplantations. The graft functional survival time (GFST) was recorded. Peripheral blood was obtained two weeks after the transplantations to prepare single cell suspension for detecting chimera formation rate and the level of Vbeta11+ T cell by flow cytometry. RESULTS: The average GFST of group IV was (18 +/- 2.2) d, significantly longer than those of group I (7.8 +/- 1.2) d, group II (8.2 +/- 1.6) d, and group III (8.8 +/- l.4) d (P < 0.05). The rats in group IV had significant lower level of Vbeta11+ T cells (2.5 +/- 0.3)% than those in the other groups (P < 0.05). Donors' bone marrow-derived cells could be detected in the peripheral blood of diabetic rats in group IV, with a chimeric rate of (10.0 +/- 2.3)%. No donors' bone marrow-derived cells were detected in the rats in other groups. CONCLUSION: Bone marrow cell transplantation combined with short-term use of cyclophosphamide promote chimerism formation and induce immune tolerance in rats with pancreatic transplantations, which prolongs pancreatic graft functional survival time.

The alteration of protein profile of Walker 256 carinosarcoma cells during the apoptotic process induced by ultrasound.

The objective of this study was to investigate the alteration of the protein profile in cells after sonication and to identify the key proteins involved in the process of cell apoptosis. Walker 256 carinosarcoma cells were exposed to focused ultrasound (US) at the intensity of 2.0, 7.0, 10.2, 14.2 and 17.0 W/cm2 (I(spta)) for 10 min in vitro and the morphologic and functional changes of the cells were detected by hematoxylin & eosin staining and flow cytometry, with double staining of fluorescein isothiocyanate (FITC)-labeled Annexin V/propidium iodide (PI). The protein compositions in the cells after sonication were detected by 2-D SDS polyacrylamide gel electrophoresis. Our results showed that apoptosis of Walker 256 carinosarcoma cells could be induced by US. The percentage of early apoptosis and secondary necrosis increased with increasing intensity of US irradiation. Comparing with the protein patterns of cells before sonication, it was found that around 420 new protein spots were present in the gel after sonication. Among them, Hsp60 and Bcl-2 like protein 13 were found to be involved in the process of cell apoptosis and US-induced apoptosis of the cells was probably performed through the pathway of promoting the activation of caspase-3.