Inheritance of natural seed-coat cracking in chickpea.

A spontaneous mutant with natural seed-coat cracking, designated "cracked seed-coat mutant (CSM)," was identified in chickpea (Cicer arietinum L.) from an F(2) population of a cross ICRISAT chickpea (ICC) 10301 × ICC 12430. The extent of seed-coat cracking (SCC) varied widely from a minute to several wide cracks. Seed coats showed cracks before seeds were fully developed and the plants had reached physiological maturity. However, seed-coat cracks were most visible on dry matured seeds, particularly in desi types. Two loci (Scc-1 and Scc-2) that controlled SCC were identified. F(1) plants from the crosses of CSM with desi genotypes produced seeds with no SCC, whereas F(1) plants from the crosses of CSM with kabuli genotypes produced seeds with SCC. F(2) segregation followed 13:3 and 7:9 ratios for plants without SCC and with SCC in CSM × Desi and CSM × Kabuli crosses, respectively. Three alleles were identified at the first locus (Scc-1) from CSM (Scc-1 (c)), desi (Scc-1 (d)), and kabuli (Scc-1 (k)) types, with the dominance relationship being Scc-1 (d) > Scc-1 (c) > Scc-1 ( k ). At the second locus (Scc-2), CSM had the dominant allele (Scc-2), whereas both desi and kabuli types had the recessive allele (scc-2). The SCC trait showed no linkage with leaf type (pinnate vs. simple) and flower color (pink vs. white) and had no adverse effects on grain yield. The SCC trait may facilitate dehulling and preparation of splits (dal), but the cracked seed would be prone to damage by insect pests and unfavorable moisture conditions.

Expression and localization of urokinase-type plasminogen activator in human spinal column tumors.

We have sought to determine the production and activity of serine proteases in primary and metastatic spinal tumors and the association of these enzymes with the invasive and metastatic properties of spinal column tumors. Using immunohistochemical techniques, the cellular localization and expression of urokinase-type plasminogen activator (uPA) was assessed, whereas its activity was determined by fibrin zymography, and the amounts of enzyme were measured by an enzyme-linked immunosorbent assay (ELISA) in primary spinal column tumors (chordoma, chondrosarcoma, and giant cell tumor) and metastatic tumors of the spine arising from various malignancies (breast, lung, thyroid, and renal cell carcinomas, and melanomas). Metastatic tumors displayed higher levels of uPA activity than did primary spinal tumors (P<0.001). Immunohistochemical analysis revealed that uPA expression was highest in metastases from lung and breast carcinomas and melanomas, followed by metastatic tumors from thyroid and renal cell carcinomas. Similar results were obtained for uPA activity and enzyme level as determined by fibrin zymography and ELISA, respectively. We conclude that metastatic spinal tumors possess higher levels of uPA expression and activity than the primary spinal tumors, which tend to be less aggressive and only locally invasive malignancies. The results suggest that the plasminogen system may participate in the metastasis of tumors to the spinal column.

Expression and role of matrix metalloproteinases MMP-2 and MMP-9 in human spinal column tumors.

Matrix metalloproteinases (MMPs) have been implicated in the process of tumor invasion and metastasis formation. Thus, we determined the expression of MMPs in various primary and metastatic spinal tumors in order to assess the role of these enzymes in spinal invasion. MMP expression was examined by immunohistochemical localization, and quantitative evaluation of MMP protein content was determined by enzyme-linked immunosorbant assay (ELISA) and Western blotting. MMP enzyme activity was determined by gelatin zymography. Lung carcinomas and melanomas metastatic to the spine were shown to have higher levels of MMP-9 activity than those of breast, thyroid, renal metastases and primary spinal tumors. Immunohistochemical analysis revealed similar difference in expression of MMP-9 in tissue samples. When the tissue samples were subjected to gelatin zymography for examination of MMP-2 and MMP-9 activity and to ELISA and Western blotting for quantitative estimation of protein content, the most striking results were obtained for lung carcinomas and melanomas relative to the other tumors. Lung carcinomas and melanomas metastatic to the spine had considerably higher levels of MMP-9 activity than those of primary spinal tumor or breast, thyroid, and renal carcinoma metastases. Within the metastatic tumor category, neoplasms that are known to be associated with the shortest overall survival rates and most aggressive behavior, such as lung carcinomas and melanomas, had the highest levels of MMP-2 and MMP-9 activity compared to those less aggressive metastatic tumors such as breast, renal cell, and thyroid carcinomas. Our results suggest that MMPs may contribute to the metastases to the spinal column, and overexpression of these enzymes may correlate with enhanced invasive properties of both primary and metastatic spinal tumors.