Chemotherapeutic deletion of CTG repeats in lymphoblast cells from DM1 patients

Myotonic dystrophy type 1 (DM1) is caused by the expansion of a (CTG).(CAG) repeat in the DMPK gene on chromosome 19q13.3. At least 17 neurological diseases have similar genetic mutations, the expansion of DNA repeats. In most of these disorders, the disease severity is related to the length of the repeat expansion, and in DM1 the expanded repeat undergoes further elongation in somatic and germline tissues. At present, in this class of diseases, no therapeutic approach exists to prevent or slow the repeat expansion and thereby reduce disease severity or delay disease onset. We present initial results testing the hypothesis that repeat deletion may be mediated by various chemotherapeutic agents. Three lymphoblast cell lines derived from two DM1 patients treated with either ethylmethanesulfonate (EMS), mitomycin C, mitoxantrone or doxorubicin, at therapeutic concentrations, accumulated deletions following treatment. Treatment with EMS frequently prevented the repeat expansion observed during growth in culture. A significant reduction of CTG repeat length by 100-350 (CTG).(CAG) repeats often occurred in the cell population following treatment with these drugs. Potential mechanisms of drug-induced deletion are presented.     

Morphological changes in the rat exocrine pancreas after pancreatic duct ligation

In the present study, morphological changes of the exocrine pancreas in rats after pancreatic duct ligation were examined with light microscopy (hematoxylin-eosin, TUNEL, and PCNA staining) and scanning electron microscopy in order to elucidate the effects of increased pancreatic duct pressure. On the fifth day after pancreatic duct ligation, ductular proliferation, periductal fibrosis, and disappearance of acini were observed. TUNEL and PCNA staining demonstrated many apoptotic acinar cells and proliferating ductal cells immediately after ligation, which reached a maximal number on the 2nd or 3rd day. Tortuous or helical interlobular pancreatic ducts with inner surfaces containing many crater-like depressions and long cilia were found after ligation. These changes were almost identical to those observed in the pancreatic tissue of model chronic pancreatitis rats, WBN/Kob rats, and stroke-prone spontaneously hypertensive (SHRSP) rats. In summary, the morphological changes observed after pancreatic duct ligation were similar to those of chronic pancreatitis, therefore, the characteristic changes of pancreatic ducts observed in chronic pancreatitis may be caused by increased pancreatic duct pressure.     

Mapping of the QTL (quantitative trait locus) conferring partial resistance to leaf blast in rice cultivar Chubu 32

The rice cultivar Chubu 32 possesses a high level of partial resistance to leaf blast. The number and chromosomal location of genes conferring this resistance were detected by restriction fragment length polymorphism (RFLP) linkage mapping and quantitative trait locus (QTL) analysis. For the mapping, 149 F₃ lines derived from the cross between rice cultivar Norin 29, with a low level of partial resistance, and Chubu 32 were used, and their partial resistance to leaf blast was assessed in upland nurseries. A linkage map covering six chromosomes and consisting of 36 RFLP markers was constructed. In the map, only one significant QTL (LOD>2.0) for partial resistance was detected on chromosome 11. This QTL explained 45.6% of the phenotypic variation. The segregation ratio of the F3 lines was 3:1 for partial resistance to susceptibility. These results suggest that the partial resistance in Chubu 32 is controlled by a major gene. Received: 15 March 2001 / Accepted: 13 August 2001     

A bout of resistance exercise increases urinary calcium independently of osteoclastic activation in men

Ashizawa, Noriko, Rei Fujimura, Kumpei Tokuyama, andMasashige Suzuki. A bout of resistance exercise increases urinary calcium independently of osteoclastic activation in men.J. Appl. Physiol. 83(4):1159-1163, 1997.Metabolic acidosis increases urinary calciumexcretion in humans as a result of administration of ammonium chloride,an increase in dietary protein intake, and fasting-inducedketoacidosis. An intense bout of exercise, exceeding aerobic capacity, also causes significant decrease in blood pH as aresult of increase in blood lactate concentration. In this study weinvestigated changes in renal calcium handling, plasma parathyroidhormone concentration, and osteoclastic bone resorption after a singlebout of resistance exercise. Ten male subjects completed about of resistance exercise with an intensity of 60% of one repetitionmaximum for the first set and 80% of one repetition maximum for thesecond and third sets. After exercise, blood and urine pH shiftedtoward acidity and urinary calcium excretion increased.Hypercalciuria was observed in the presence of an increased fractionalcalcium excretion and an unchanged filtered load of calcium. Therefore,the observed increase in urinary calcium excretion was due primarily todecrease in renal tubular reabsorption of calcium. Likely causes of theincrease in renal excretion of calcium are metabolic acidosis itselfand decreased parathyroid hormone. When urinary calcium excretionincreased, urinary deoxypyridinoline, a marker of osteoclastic boneresorption, decreased. These results suggest that1) strenuous resistance exerciseincreased urinary calcium excretion by decreasing renal tubular calciumreabsorption, 2) urinary calciumexcretion increased independently of osteoclast activation, and3) the mechanism resulting inpostexercise hypercalciuria might involve non-cell-mediatedphysicochemical bone dissolution.

     

Characterization of a YAC and cosmid contig containing markers tightly linked to the myotonic dystrophy locus on chromosome 19.

Myotonic dystrophy (DM) is caused by a defect in an unknown gene that maps to 19q13.3, flanked by the tightly linked markers ERCC1 on the proximal side and D19S51 on the distal side. We report the isolation and characterization of overlapping YAC and cosmid clones around D19S51 for the construction of a physical map around this locus. The resulting contig contains the markers D19S51 and D19S62 (another new marker tightly linked to the DM locus) and the distal breakpoint of a radiation hybrid cell line used in the physical mapping of the DM region. We have compared the restriction maps of the YACs and cosmids with that of the genome to investigate the fidelity of these clones.     

Regulation of flagellar motility by temperature-dependent phosphorylation of a 43 kDa axonemal protein in fowl spermatozoa.

Phosphorylation of fowl sperm proteins was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) after incubating the demembranated spermatozoa with [gamma-32P]ATP at 30 degrees C or 40 degrees C. A marked difference of phosphorylation between 30 degrees C and 40 degrees C was observed in a 43 kDa protein. This protein was slightly phosphorylated at 40 degrees C, but strongly phosphorylated at 30 degrees C in a cAMP-independent manner. The motility of demembranated spermatozoa was negligible at 40 degrees C, but vigorous movement was observed at 30 degrees C. These results showed that phosphorylation of a 43 kDa protein is likely to be a regulatory step in the maintenance of fowl sperm motility.     

Hypoxia within macrophyte vegetation limits the use of methane-derived carbon by larval chironomids in a shallow temperate eutrophic lake

Methane-derived carbon (MDC) can subsidize lake food webs. However, the trophic transfer of MDC to consumers within macrophyte vegetation is largely unknown. We investigated the seasonality of δ¹³C in larval chironomids within Nelumbo nucifera (Gaertn.) and Trapa natans var. Japonica (Nakai) vegetation in the shallow, eutrophic Lake Izunuma in Japan. Over the past several years, N. nucifera has rapidly expanded across more than 80% of the lake surface. Prior to the expansion of N. nucifera (2007–2008), a previous study reported extremely low larval δ¹³C levels with peak sediment methane concentrations in August or September. After the expansion of N. nucifera (2014–2015), we observed extreme hypoxia as low as or lower than 1 mg l^?1 among the macrophyte coverage during June and August. During August and September, no larvae could be found among N. nucifera, and larvae in T. natans showed relatively high δ¹³C levels (>???40‰). In contrast, larvae were markedly ¹³C–depleted (down to ??60‰) during October and November. The renewed supply of oxygen to the lake bottom may stimulate MOB activity, leading to an increase in larval assimilation of MDC. Our results suggest that macrophyte vegetation can affect the seasonality of MDC transfer to benthic consumers under hypoxic conditions in summer.     

Cardiovascular and sympathetic nerve responses induced by intracerebroventricular injections of prostacyclin in rats

Intracerebroventricular (ICV) injections of prostacyclin (PGI2) produced biphasic blood pressure responses consisting of an initial hypotensive phase followed by a sustained pressor phase in awake rats. Heart rate increased following such injections in either awake or anesthetized rats. PGI2, 1 microgram, produced biphasic responses and, 10 micrograms, purely vasodepressor responses in anesthetized rats, but abdominal sympathetic nerve firing recorded was consistently increased. Hypophysectomy did not affect the hypotensive phase of the responses. These results indicate that the initial hypotension can not be explained by centrally-induced changes in sympathetic nerve activity or vasopressin release, but may be due to peripheral effects of PGI2 leaking from the injection site.