Double-strand breaks and the concept of short- and long-term epigenetic memory

Double-strand breaks represent an extremely cytolethal form of DNA damage and thus pose a serious threat to the preservation of genetic and epigenetic information. Though it is well-known that double-strand breaks such as those generated by ionising radiation are among the principal causative factors behind mutations, chromosomal aberrations, genetic instability and carcino-genesis, significantly less is known about the epigenetic consequences of double-strand break formation and repair for carcinogenesis. Double-strand break repair is a highly coordinated process that requires the unravelling of the compacted chromatin structure to facilitate repair machinery access and then restoration of the original undamaged chromatin state. Recent experimental findings have pointed to a potential mechanism for double-strand break-induced epigenetic silencing. This review will discuss some of the key epigenetic regulatory processes involved in double-strand break (DSB) repair and how incomplete or incorrect restoration of chromatin structure can leave a DSB-induced epigenetic memory of damage with potentially pathological repercussions

Quantification of gammaH2AX foci in response to ionising radiation

DNA double-strand breaks (DSBs), which are induced by either endogenous metabolic processes or by exogenous sources, are one of the most critical DNA lesions with respect to survival and preservation of genomic integrity. An early response to the induction of DSBs is phosphorylation of the H2A histone variant, H2AX, at the serine-139 residue, in the highly conserved C-terminal SQEY motif, forming gammaH2AX(1). Following induction of DSBs, H2AX is rapidly phosphorylated by the phosphatidyl-inosito 3-kinase (PIKK) family of proteins, ataxia telangiectasia mutated (ATM), DNA-protein kinase catalytic subunit and ATM and RAD3-related (ATR)(2). Typically, only a few base-pairs (bp) are implicated in a DSB, however, there is significant signal amplification, given the importance of chromatin modifications in DNA damage signalling and repair. Phosphorylation of H2AX mediated predominantly by ATM spreads to adjacent areas of chromatin, affecting approximately 0.03% of total cellular H2AX per DSB(2,3). This corresponds to phosphorylation of approximately 2000 H2AX molecules spanning approximately 2 Mbp regions of chromatin surrounding the site of the DSB and results in the formation of discrete gammaH2AX foci which can be easily visualized and quantitated by immunofluorescence microscopy(2). The loss of gammaH2AX at DSB reflects repair, however, there is some controversy as to what defines complete repair of DSBs; it has been proposed that rejoining of both strands of DNA is adequate however, it has also been suggested that re-instatement of the original chromatin state of compaction is necessary(4-8). The disappearence of gammaH2AX involves at least in part, dephosphorylation by phosphatases, phosphatase 2A and phosphatase 4C(5,6). Further, removal of gammaH2AX by redistribution involving histone exchange with H2A.Z has been implicated(7,8). Importantly, the quantitative analysis of gammaH2AX foci has led to a wide range of applications in medical and nuclear research. Here, we demonstrate the most commonly used immunofluorescence method for evaluation of initial DNA damage by detection and quantitation of gammaH2AX foci in gamma-irradiated adherent human keratinocytes(9)

Methylproamine protects against ionizing radiation by preventing DNA double-strand breaks

Purpose The majority of cancer patients will receive radiotherapy (RT), therefore, investigations into advances of this modality are important. Conventional RT dose intensities are limited by adverse responses in normal tissues and a primary goal is to ameliorate adverse normal tissue effects. The aim of these experiments is to further our understanding regarding the mechanism of radioprotection by the DNA minor groove binder, methylproamine, in a cellular context at the DNA level. Materials and methods We used immunocytochemical methods to measure the accumulation of phosphorylated H2AX (γH2AX) foci following ionizing radiation (IR) in patient-derived lymphoblastoid cells exposed to methylproamine. Furthermore, we performed pulsed field gel electrophoresis DNA damage and repair assays to directly interrogate the action of methylproamine on DNA in irradiated cells. Results We found that methylproamine-treated cells had fewer γH2AX foci after IR compared to untreated cells. Also, the presence of methylproamine decreased the amount of lower molecular weight DNA entering the gel as shown by the pulsed field gel electrophoresis assay. Conclusions These results suggest that methylproamine acts by preventing the formation of DNA double-strand breaks (dsbs) and support the hypothesis that radioprotection by methylproamine is mediated, at least in part, by decreasing initial DNA damage.

Impacts of pesticides and fertilisers on soil

The effects of inorganic amendments (fertilisers and pesticides) on soil biota that are reported in the scientific literature are, to say the least, variable. Though there is clear evidence that certain products can have significant impacts, the effects can be positive or negative. This is not surprising when you consider the number of organisms and amount of different functional groups, the number of products and various rates at which they could be applied, the methods of application and the environmental differences that occur in soil at a micro scale (within centimetres) in a paddock, let alone between paddocks, farms, catchments, regions etc. It therefore becomes extremely difficult to draw definitive conclusions from the reported results in order to summarise the impacts of these inputs. Several research trials and review papers have been published on this subject and most similarly conclude that the implications of many of the effects are still uncertain.

Lithology and palynology of cave floor sediment cores from Wakulla Spring, Wakulla County, Florida

Five short bottom sediment cores taken in Wakulla Spring Wakulla County, Florida, were described lithologically and sampled for palynological study. Four of the cores were recoveredfrom sediments at the spring cave entrance (130 feet water depth). One core was taken in a fossil vertebrate bone bed, 280 feet distance into the main spring cave at a water depth of 240 feet. Sediments in the cores are composed of alternating intervals of quartz sand and calcilitite, containing freshwater diatoms, freshwater mollusk shells and plant remains. The predominant pollen present in all cores consists of a periporate variety typical of the herb families Chenopodiaceae and Amaranthaceae. Arboreal flora, typical of the area surrounding the spring today, represent a very low percentage of thle pollen assemblage in the cores. Clustered Chenopod-Amaranth type pollen observed in one core suggest minimal transport prior to deposition, and indicate that the bottom sediments in the cave may be essentially In situ. An absence of exotic flora suggests a Quaternary age for the sediments. (PDF contains 11 pages.)

Miscellaneous studies

MISCELLANEOUS STUDIES, which includes the following papers: "Geology of the Area in and Around the Jim Woodruff Reservoir" by Charles W. Hendry, Jr. and J. William Yon, Jr.; "Phosphate Concentrations near Bird Rookeries in South Florida" by Dr. Ernest H. Lund, Department of Geology, Florida State University; and "An Analysis of Ochlockonee River Channel Sediments" by Dr. Ernest H. Lund, Associate Professor and Patrick C. Haley, Graduate Assistant, Department of Geology, Florida State University. (PDF contains 81 pages)

Alligator Diet in Relation to Alligator Mortality on Lake Griffin, FL

Alligator mississippiensis (American Alligators) demonstrated low hatchrate success and increased adult mortality on Lake Griffin, FL, between 1998 and 2003. Dying Lake Griffin alligators with symptoms of poor motor coordination were reported to show specific neurological impairment and brain lesions. Similar lesions were documented in salmonines that consumed clupeids with high thiaminase levels. Therefore, we investigated the diet of Lake Griffin alligators and compared it with alligator diets from two lakes that exhibited relatively low levels of unexplained alligator mortality to see if consumption of Dorosoma cepedianum (gizzard shad) could be correlated with patterns of mortality. Shad in both lakes Griffin and Apopka had high levels of thiaminase and Lake Apopka alligators were consuming greater amounts of shad relative to Lake Griffin without showing mortality rates similar to Lake Griffin alligators. Therefore, a relationship between shad consumption alone and alligator mortality is not supported.

Long-term natural acidification of upland sites in Cumbria: Evidence from post-glacial lake sediments

Studies by the Freshwater Biological Association over the last 25 years have supplied data relevant to the levels of acidity in local soils and water before the onset of industrial pollution and current interest in acid rain. This article reviews published analysis from cores of lake sediments, in or near the catchment of the River Duddon. Electron spin resonance spectra of humic acids and iodine values confirm evidence from pollen analysis for a history of progressive acidification of the source material of lake sediments since before 5000 radiocarbon years, in upland catchments of the Lake District. Processes involved included: removal of basic ions from soils by rainfall, the effects of which were intensified by removal by man of deciduous forest; acidification of soils and waters by decomposition products of Calluna and further acidification of waters by Sphagnum species which colonized habitats where drainage became impeded by paludification processes.

Der Befall von Seelachsfilets mit Nematodenlarven und deren Abtötung durch Tiefgefrieren

Im Rahmen unserer Untersuchungen zum Befall von Seefischen mit Nematodenlarven und deren Abtötung unter industriellen Verarbeitungsbedingungen wurden während der 88. Reise des FFS "Walther Herwig" vom 23.1.88-17.2.88 u.a. Seelachs (Köhler; Pollachius virens) verschiedener Größenklassen untersucht.