Site-specific integration of an F'' lac pro factor in the region of the replication origin (oriC) of E. coli

Summary An episome, F 128, which carries approximately 8x10⁴ base pairs of chromosomal DNA homologous to the lac pro region of the E. coli chromosome, has been found to integrate into the oriC region of the chromosome in a site specific reaction. While the event appears to be recA-dependent, no homology between the episome and this region of the chromosome was detected. The Hfr strains formed result from the integration of intact F 128 molecules. The structure of the Hfr strains generated has been determined and their transfer properties analyzed.     

The ultrastructure of the symbionts ofRhizocarpon geographicum,Parmelia conspersa andUmbilicaria pustulata growing under dryness conditions

Summary The dryness-induced ultrastructural changes of both myco- and phycobiont of three lichen species (R. geographicum, P. conspersa, andU. pustulata) have been studied over three months and half, period of time. During this time other ecological factors, such as rock substratum, temperature, light and gas interchange were unaltered compared to the natural conditions. A large number of ultrastructural changes were observed in the mycobiont as well as in the phycobiont (Trebouxia) and often, cells showed a highly disorganized morphology. The most important ultrastructural modifications were: 1. pyrenoglobuli of the algae were peripheral, 2. new and unknown structures were observed in the phycobionts of bothR. geographicum andU. pustulata as well as in the mycobiont of the latter species.     

Versatility and specialization in labrid fishes: ecomorphological implications

Summary The term specialized has been used to describe species that possess unique functional attributes and/or a narrow, stereotyped range of attributes, but there are few comparative functional analyses of specialists and generalists. If species with functional morphological specializations are capable of functioning over a broad range, the link between morphology and ecology may be relaxed under certain environmental conditions. In this study, high-speed films of jaw movements during prey capture were compared statistically for three coexisting coral reef fish species in the family Labridae, one trophic specialist and two trophic generalists. The trophic specialist possessed a unique functional feature related to the movement of the hyoid in the floor of the mouth, while the trophic generalists were not observed to possess any functional specializations. All three species showed functional versatility in that they were able to adjust their prey capture mechanism in response to the evasive potential of the prey. The functional versatility of trophic specialists has implications for ecomorphological studies, since species characterized as possessing unique functional or morphological features may demonstrate marked flexibility in ecological variables such as diet or foraging behavior, decreasing the likelihood of identifying correlations between morphology and ecology.     

The distribution of RFLP markers on chromosome 2(2H) of barley in relation to the physical and genetic location of 5S rDNA

The 5S rDNA locus on the long arm of barley chromosome 2(2H) was genetically mapped in two crosses in relation to 30 other RFLP loci. Comparison of the genetic maps with the previously published physical position of the 5S rDNA, determined by in-situ hybridization, showed that there was a marked discrepancy between physical and genetic distance in both crosses, with recombination being less frequent in the proximal part of the arm. Pooled information from the present study and other published genetic maps showed that at least 26 of the 44 (59%) RFLPs that have been mapped on 2(2H)L lie distal to the 5S rDNA locus even though this region is only 27% of the physical length of the arm. The distribution of RFLP markers is significantly different from expected (P < 0.01), implying that the low-copy sequences used for RFLP analysis occur more frequently in distal regions of the arm and, or, that sequences in distal regions are more polymorphic.     

Variation of cooling rate and concentration of dimethyl sulfoxide on rabbit kidney function

Rabbit kidney function was assessed in vitro after cryoprotection with either 3 or 4 M dimethyl sulfoxide. The introduction and removal of the cryoprotectant was carried out in a stepwise progressive manner and the removal in a stepwise progressive manner with hypertonic mannitol solutions. This in vitro model can be shown to respond to various ischemic-like states resulting in poor or absent function. Active tubular transport can be demonstrated. It has been used by many authors as an intermediate step prior to the ultimate test of reimplant and contralateral nephrectomy. Variations in the rate of cooling at cryoprotection levels of 3 and 4 M dimethyl sulfoxide concentration (Me2SO) were carried out. In general, at 3 M concentration of Me2SO, creatinine clearance, sodium and glucose reabsorption are preserved with a fair degree of success after cooling to -10, -15, and -20 degrees C in our model, when the rate of cooling to these levels is 1.0 degree C/min. When a cooling rate of 0.5 degree C/min is used, renal function is significantly reduced whether the final temperature is -10, -15, or -20 degrees C. Control rabbit kidneys will tolerate 4 M concentration of Me2SO and give fairly good function. When cooled to -15 or -20 degrees C, there is poor function at 0.1 and 0.5 degrees C/min. Fair function is obtained at the rate of 1 degree C/min to -10 degrees C. Therefore, at cryoprotectant levels of 3 and 4 M Me2SO, kidney function as assayed by in vitro perfusion, is better when the cooling rate is 1.0 degree C/min.     

Premortem autophagy determines the immunogenicity of chemotherapy-induced cancer cell death

One particular strategy to render anticancer therapies efficient consists of converting the patient's own tumor cells into therapeutic vaccines, via the induction of immunogenic cell death (ICD). One of the hallmarks of ICD dwells in the active release of ATP by cells committed to undergo, but not yet having succumbed to, apoptosis. We observed that the knockdown of essential autophagy-related genes (ATG3, ATG5, ATG7 and BECN1) abolishes the pre-apoptotic secretion of ATP by several human and murine cancer cell lines undergoing ICD. Accordingly, autophagy-competent, but not autophagy-deficient, tumor cells treated with ICD inducers in vitro could induce a tumor-specific immune response in vivo. Cancer cell lines stably depleted of ATG5 or ATG7 normally generate tumors in vivo, and such autophagy-deficient neoplasms, upon systemic treatment with ICD inducers, exhibit the same levels of apoptosis (as monitored by nuclear shrinkage and caspase-3 activation) and necrosis (as determined by following the kinetics of HMGB1 release) as their autophagy-proficient counterparts. However, autophagy-incompetent cancers fail to release ATP, to recruit immune effectors into the tumor bed and to respond to chemotherapy in conditions in which autophagy-competent tumors do so. The intratumoral administration of ecto-ATPase inhibitors increases extracellular ATP concentrations, re-establishes the therapy-induced recruitment of dendritic cells and T cells into the tumor bed, and restores the chemotherapeutic response of autophagy-deficient cancers. Altogether, these results suggest that autophagy-incompetent tumor cells escape from chemotherapy-induced (and perhaps natural?) immunosurveillance because they are unable to release ATP.