Calcium and EGTA Alleviate Cadmium Toxicity in Germinating Chickpea Seeds

Impact of exogenous calcium and ethylene glycol tetraacetic acid (EGTA) supplement on chickpea (Cicer arietinum L.) germinating seeds exposed to cadmium stress for 6 days was studied. Ca and EGTA late treatment (3 days) alleviated growth inhibition and decreased Cd accumulation as well as lipid peroxidation and protein carbonylation in both root and shoot cells. Exogenous effector application relieved Cd-induced cell death which was associated with a constant level of ATP, which was considered as an apoptotic-like process. Redox balance was examined through the study of the redox state of pyridine nucleotide couples NAD⁺/NADH and NADP⁺/NADPH as well as their related oxidative [NAD(P)H-oxidase] and dehydrogenase (glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and malate dehydrogenase) enzyme activities. The present research illustrated an ameliorative effect of Ca and EGTA on growth of Cd-exposed chickpea seedlings that occurs through the protection of sensitive cell sites from Cd-induced oxidation, namely membrane lipids and proteins, rather than the improvement of recycling capabilities of the cellular reducing power.     

Chemical Constituents in Hybrids of Ligularia tongolensis and L. cymbulifera: Chemical Introgression in L. tongolensis

Two samples with morphologies intermediate between Ligularia tongolensis and L. cymbulifera were collected in Desha, Sichuan Province, and one, in Pachahai, Yunnan Province, P. R. China. The DNA sequencing confirmed that the samples were hybrids of the two species. Tetradymol ( 1 ), the major compound of L. cymbulifera not found in L. tongolensis, was isolated from the hybrid samples collected at both locations, while furanoeremophilan‐15‐oic acid derivative 4 , a compound characteristic to L. tongolensis, was found in the Pachahai hybrid but not in the Desha hybrids. Thus, the chemical consequence of hybridization can be variable. In addition, analysis of L. tongolensis samples at Pachahai indicated that introgression has been a mechanism of generating chemical diversity in the plant. Eleven compounds including three new ones were isolated.     

The Efficacy of Continued Sorafenib Treatment after Radiologic Confirmation of Progressive Disease in Patients with Advanced Hepatocellular Carcinoma

Background

Whether radiologically detected progressive disease (PD) is an accurate metric for discontinuing sorafenib treatment in patients with hepatocellular carcinoma (HCC) is unclear. We investigated the efficacy of sorafenib treatment after radiologic confirmation of PD in patients with advanced HCC.

Methods

We retrospectively analyzed HCC patients treated with sorafenib at Kyushu Medical Center. Six of the 92 patients with radiologically confirmed PD were excluded because they were classified as Child-Pugh C or had an Eastern Cooperative Oncology Group (ECOG) performance status (PS) ≥3; 86 patients were ultimately enrolled.

Results

Among the 86 patients, 47 continued sorafenib treatment after radiologic confirmation of PD (the continuous group), whereas 39 did not (the discontinuous group). The median survival time (MST) in the continuous group after confirmation was 12.9 months compared with 4.5 months in the discontinuous group (p <0.01). The time to progression in the continuous group after confirmation was 2.6 months compared with 1.4 months in the discontinuous group (p <0.01); it was 4.2 months and 2.1 months in patients who had received sorafenib ≥4 months and <4 months, respectively, before confirmation (p = 0.03). In these subgroups, the post-PD MST was 16.7 months and 9.6 months, respectively (p < 0.01). Independent predictors of overall survival after radiologic detection of PD were (hazard ratio, confidence interval): ECOG PS <2 (0.290, 0.107–0.880), Barcelona Clinical Liver Cancer stage B (0.146, 0.047–0.457), serum α-fetoprotein level ≥400 ng/mL (2.801, 1.355–5.691), and post-PD sorafenib administration (0.279, 0.150–0.510).

Conclusion

Continuing sorafenib treatment after radiologic confirmation of PD increased survival in patients with advanced HCC. Therefore, radiologically detected PD is not a metric for discontinuation of sorafenib treatment in such patients.     

On the Utility of MIBG SPECT/CT in Evaluating Cardiac Sympathetic Dysfunction in Lewy Body Diseases

BackgroundAbnormal cardiac uptake of ¹²³I-metaiodobenzylguanidine (¹²³I-MIBG) is a diagnostic marker of Lewy body diseases (LBDs), e.g., Parkinson’s disease (PD) and dementia with Lewy bodies (DLB). Planar imaging is generally used to assess cardiac sympathetic dysfunction in ¹²³I-MIBG scintigraphy; however, its clinical utility requires further improvement. We hypothesized that the co-registration of single-photon emission tomography (SPECT) and computed tomography (CT) images would improve the diagnostic accuracy of ¹²³I-MIBG cardiac scintigraphy for LBDs. This study sought to evaluate the effects of SPECT/CT imaging on ¹²³I-MIBG cardiac scintigraphy for diagnosing LBDs.MethodsWe retrospectively investigated data of 54 patients (consecutive 18 patients in each PD, DLB, and idiopathic normal pressure hydrocephalus [iNPH] groups) who underwent ¹²³I-MIBG cardiac scintigraphy (planar and SPECT/CT) because of suspected LBDs at the Tohoku University hospital from June 2012 to June 2015. We compared the diagnostic accuracies of the conventional planar ¹²³I-MIBG method and SPECT/CT methods (manual and semi-automatic).ResultsIn the conventional planar analysis, ¹²³I-MIBG uptake decreased only in the DLB group compared with the iNPH group. In contrast, the SPECT/CT analysis revealed significantly lower ¹²³I-MIBG uptake in both the PD and DLB groups compared with the iNPH group. Furthermore, a receiver operating characteristic analysis revealed that both the manual and semi-automatic SPECT/CT methods were superior to the conventional planar method in differentiating the 3 disorders.ConclusionsSPECT/CT ¹²³I-MIBG cardiac scintigraphy can detect mild cardiac sympathetic dysfunction in LDBs. Our results suggest that the SPECT/CT technique improves diagnostic accuracy for LBDs.     

A Pressure-Jump Method as a New Tool in Growth Physiology for Monitoring Physiological Wall Extensibility and Effective Turgor

A new method has been devised in order to allow the rapid determinationof the physiological extensibility () of the cell wall togetherwith the apparent effective turgor (P — Y'). The methodconsists of determining the elongation growth rate of a segmentof pea hypocotyl with an auxanometer and applying a very smallincrease (around 10 kPa) in xylem pressure (jumps) by the xylemperfusion method. The cell turgor pressure is increased by thewater surge from the xylem that is due to the pressure jumpand reaches a new level within a short time. The elongationrate is also increased asymptotically to a new steady-statelevel within a short time. is estimated by simply dividingthe increase in the steady-state growth rate by the increasein perfusion pressure, which is monitored by a pressure transduceror by the increase in turgor pressure measured by a pressureprobe. Minimal length of the time period required to get a newsteady elongation rate is 1.5 min. Therefore, it is possibleto scan continuously the change in by the intermittent applicationof the brief, small pulses of xylem pressure during any experimentaltreatment. The apparent effective turgor P — Y' can becalculated by dividing the initial elongation growth rate by Using this method, we have demonstrated experimentally thenon-linearity of the relationship between elongation rate andturgor (Lockhart mechanical equation). (Received April 26, 1989; Accepted July 17, 1989)     

Molecular nature of chromosome 5q loss in colorectal tumors and desmoids from patients with familial adenomatous polyposis

Summary Familial adenomatous polyposis (FAP), which includes familial polyposis coli (FPC) and the Gardner syndrome (GS), is a genetically determined premalignant disease of the colon inherited by a locus (APC) mapping within 5q15–q22. To elucidate the role of 5q loss in FAP tumorigenesis, we analysed 51 colorectal tumors and seven desmoids from 19 cases of FPC and five GS patients, as well as 15 sporadic colon cancers. RFLP analysis revealed a high incidence of allelic deletion in hereditary colon cancers as well as in sporadic colon cancers with a peak at the APC locus. APC loss resulted primarily from interstitial deletion or mitotic recombination. Combined tumor and pedigree analysis in a GS family revealed loss of normal 5q alleles in three tumors, including a desmoid tumor, which suggests the involvement of hemizygosity or homozygosity of the defective APC gene in colon carcinogenesis and, possibly, in extracolonic neoplasms associated with FAP.     

β-Galactosidase-deficient human fibroblasts: uptake and processing of the exogenous precursor enzyme expressed by stable transformant COS cells

Summary COS-1 cells were transfected by electroporation with a cDNA for human acid -galactosidase cloned in our laboratory and stable transformants expressing the enzyme activity were selected. The precursor form of the enzyme was secreted in large quantities into the culture medium. The fibroblasts from patients with G_M1-gangliosidosis or Morquio B disease showed a remarkable increase of enzyme activity, up to the normal level, after culture in this medium for 2 days; the amount of uptake was essentially the same as that for the precursor form in human fibroblasts. After endocytosis, the precursor molecules were processed normally to the mature form and remained as stable as those produced by human fibroblasts. On the other hand, cells from galactosialidosis patients did not show any increase of enzyme activity in a similar experiment. It was concluded that the transformants are useful as the source of precursor proteins for the study of intracellular turnover of enzyme molecules in mutant cells.     

Requirement of heat and metabolic energy for the expression of inhibitory action of colicin K.

Escherichia coli B, induced for beta-galactoside permease, can accumulate thio-methyl-beta-galactoside in the cell even at 0 degrees D. At this temperature, cells adsorb colicin K but the adsorbed colicin does not inhibit thiomethyl-beta-galactoside uptake. Inhibition by colicin K is, however, seen at 0 degrees C after exposure of the colicin K-cell complex to a high temperature: a greater degree of inhibition occurs with increasing temperature or duration or exposure. There is a transition point at around 21 degrees C in Arrhenius plots of this colicin K activation reaction. If inhibitors of energy yielding reactions are present during the heat treatment, the inhibitory action of colicin K (as measured by thiomethyl-beta-galactoside uptake after returning the colicin K-cell complex to 0 degrees C and removal of the inhibitors) is prevented. These results indicate that adsorbed colicin K is converted into the active state only in the presence of metabolic energy and that cell surface fluidity appears to be concerned in this process.     

Tetrodotoxin-sensitive uptake of ions and water by slices of rat brain in vitro

1. Exchange of (3)H(2)O with H(2)O takes place rapidly in incubated rat brain slices but at a lower rate in slices from infant brain than from adult brain. The temperature coefficient (Q(10)) of the exchange process, between 37 and 4 degrees C, is 1.76 with infant brain and 1.26 with adult brain. The exchange process is unaffected by the presence of ouabain or 2,4-dinitrophenol. 2. An approximately linear relationship exists between water uptake and the concentration of ATP in the incubated slices in the presence of various concentrations of glucose. Little or no change occurs in water uptake and ATP concentration in the presence of a glucose concentration exceeding 3mm. A linear relationship also exists between the water uptake and ATP concentration in the presence of 10mm-glucose and various concentrations of sodium l-glutamate but the line is parallel to that found with changed glucose concentrations and shifted in the direction of increased water uptake. A similar parallel relationship exists between water uptake and ATP concentration in the presence of 2,4-dinitrophenol, but the amount of water uptake is significantly smaller in the presence of 2,4-dinitrophenol than in its absence. 3. Copper chloride (0.3mm) or mercuric chloride (0.3mm) both increase water uptake and diminish the ATP concentration in slices. Sodium malonate (2mm) or sodium d-glutamate (10mm) has similar effects. 4. Substances, or conditions, affecting water uptake in incubated brain slices may be divided roughly into two classes in accordance with their effects on adenosine triphosphatase and membrane permeability, but there may be considerable lack of specificity.