Date Palm DNA Mini-Preparation without Liquid Nitrogen

We have developed a new mini-procedure for isolation of total cellular DNA from date palm (Phoenix dactylifera L.). The procedure, which does not use liquid nitrogen, has proved useful due to temporary disruptions in supplies of liquid nitrogen that occur in countries where date palm trees are cultivated. DNA suitable for RFLP and PCR analyses is obtained.     

Burn Injury Reduces Neutrophil Directional Migration Speed in Microfluidic Devices

Thermal injury triggers a fulminant inflammatory cascade that heralds shock, end-organ failure, and ultimately sepsis and death. Emerging evidence points to a critical role for the innate immune system, and several studies had documented concurrent impairment in neutrophil chemotaxis with these post-burn inflammatory changes. While a few studies suggest that a link between neutrophil motility and patient mortality might exist, so far, cumbersome assays have prohibited exploration of the prognostic and diagnostic significance of chemotaxis after burn injury. To address this need, we developed a microfluidic device that is simple to operate and allows for precise and robust measurements of chemotaxis speed and persistence characteristics at single-cell resolution. Using this assay, we established a reference set of migration speed values for neutrophils from healthy subjects. Comparisons with samples from burn patients revealed impaired directional migration speed starting as early as 24 hours after burn injury, reaching a minimum at 72–120 hours, correlated to the size of the burn injury and potentially serving as an early indicator for concurrent infections. Further characterization of neutrophil chemotaxis using this new assay may have important diagnostic implications not only for burn patients but also for patients afflicted by other diseases that compromise neutrophil functions.     

A comparative study of chromatin from lymphocyte nuclei upon activation of transcription by irradiation from an He-Ne-laser or phytohemagglutinin]

The influence of He-Ne laser radiation (632.8 nm, 56 J/m2, t = 10 s) and phytohaemagglutinin (PHA, 2 micrograms/ml) on chromatin structure in human lymphocytes was studied by electron microscopy using ultrathin cell sections. Morphometric analysis of extranuclear condensed chromatin masses was performed 1 h after the irradiation or after the beginning of PHA treatment. In the irradiated cells the following insignificant changes were revealed: decrease in the relative area of the nucleoplasmic chromatin, increase in the relative area of decondensation zones as well as increase in the number of clumps of nucleoplasmic chromatin and relative length at their boundary with nucleoplasma. The tendency of these morphological changes may be interpreted as functional activation of extranucleolar RNA synthesis in response to irradiation by red laser light. Action of PHA results in significant changes of the surfaces of chromatin clumps, namely increase in relative length of nucleoplasmic chromatin boundary and decrease in relative length of perimembranous chromatin boundary with nucleoplasma as well as some less expressed delamination of the chromatin masses from the nuclear membrane. These essential changes may reflect chromatin activation by proliferative stimulus. Peculiarities of the ultrastructural reorganisation in the condensed chromatin after irradiation and PHA-treatment probably reflect the differences in the processes of gene activation caused by the two agents.     

Dissociating Two Stages of Preparation in the Stop Signal Task Using fMRI

Often we must balance being prepared to act quickly with being prepared to suddenly stop. The stop signal task (SST) is widely used to study inhibitory control, and provides a measure of the speed of the stop process that is robust to changes in subjects’ response strategy. Previous studies have shown that preparation affects inhibition. We used fMRI to separate activity that occurs after a brief (500 ms) warning stimulus (warning-phase) from activity that occurs during responses that follow (response-phase). Both of these phases could contribute to the preparedness to stop because they both precede stop signals. Warning stimuli activated posterior networks that signal the need for top-down control, whereas response phases engaged prefrontal and subcortical networks that implement top-down control. Regression analyses revealed that both of these phases affect inhibitory control in different ways. Warning-phase activity in the cerebellum and posterior cingulate predicted stop latency and accuracy, respectively. By contrast, response-phase activity in fronto-temporal areas and left striatum predicted go speed and stop accuracy, in pre-supplementary motor area affected stop accuracy, and in right striatum predicted stop latency and accuracy. The ability to separate hidden contributions to inhibitory control during warning-phases from those during response-phases can aid in the study of models of preparation and inhibitory control, and of disorders marked by poor top-down control.     

Age-related variability in occlusal wear planes

Investigation of two populations of 136 individuals shows several patterns of occlusal wear plane change which are positively correlated with age. For individuals up to the age of 18, there is a characteristic pattern in which the occlusal wear planes of the mandibular teeth are lingually sloped and those of the maxillary dentition buccally sloped, with the exception of the maxillary premolars, which are also lingually sloped. The long axes of the mandibular teeth give them a lingual orientation relative to the maxillary teeth, and the long axes of the maxillary molars, by contrast, are buccally oriented. In the 18-30 age range for all sexes, the mandibular M1 becomes buccally sloped on its occlusal surface while the occlusal wear plane on the maxillary M1 becomes lingually sloped. Later age changes indicate a trend for the mandibular premolars to become buccally sloped, while the wear planes of the maxillary premolars remain lingually sloped. There is a corresponding tendency for the maxillary and mandibular second molars to undergo changes in the initial orientation of the occlusal wear planes.