Alterations in the Magnetoencephalography Default Mode Effective Connectivity following Concussion.

BACKGROUND AND PURPOSE: Magnetoencephalography is sensitive to functional connectivity changes associated with concussion. However, the directional influences between functionally related regions remain unexplored. In this study, we therefore evaluated concussion-related magnetoencephalography-based effective connectivity changes within resting-state default mode network regions. MATERIALS AND METHODS: Resting-state magnetoencephalography was acquired for 8 high school football players with concussion at 3 time points (preseason, postconcussion, postseason), as well as 8 high school football players without concussion and 8 age-matched controls at 2 time points (preseason, postseason). Time-series from the default mode network regions were extracted, and effective connectivity between them was computed for 5 different frequency bands. The default mode network regions were grouped into anterior and posterior default mode networks. The combined posterior-to-anterior and anterior-to-posterior effective connectivity values were averaged to generate 2 sets of values for each subject. The effective connectivity values were compared using a repeated measures ANOVA across time points for the concussed, nonconcussed, and control groups, separately. RESULTS: A significant increase in posterior-to-anterior effective connectivity from preseason to postconcussion (corrected P value = .013) and a significant decrease in posterior-to-anterior effective connectivity from postconcussion to postseason (corrected P value = .028) were observed in the concussed group. Changes in effective connectivity were only significant within the delta band. Anterior-to-posterior connectivity demonstrated no significant change. Effective connectivity in the nonconcussed group and controls did not show significant differences. CONCLUSIONS: The unidirectional increase in effective connectivity postconcussion may elucidate compensatory processes, invoking use of posterior regions to aid the function of susceptible anterior regions following brain injury. These findings support the potential value of magnetoencephalography in exploring directional changes of the brain network following concussion.

Prevalence and Incidence of Microhemorrhages in Adolescent Football Players.

BACKGROUND AND PURPOSE: SWI is an advanced imaging modality that is especially useful in cerebral microhemorrhage detection. Such microhemorrhages have been identified in adult contact sport athletes, and the sequelae of these focal bleeds are thought to contribute to neurodegeneration. The purpose of this study was to utilize SWI to determine whether the prevalence and incidence of microhemorrhages in adolescent football players are significantly greater than those of adolescent noncontact athletes. MATERIALS AND METHODS: Preseason and postseason SWI was performed and evaluated on 78 adolescent football players. SWI was also performed on 27 adolescent athletes who reported no contact sport history. Two separate one-tailed Fisher exact tests were performed to determine whether the prevalence and incidence of microhemorrhages in adolescent football players are greater than those of noncontact athlete controls. RESULTS: Microhemorrhages were observed in 12 football players. No microhemorrhages were observed in any controls. Adolescent football players demonstrated a significantly greater prevalence of microhemorrhages than adolescent noncontact controls (P = .02). Although 2 football players developed new microhemorrhages during the season, microhemorrhage incidence during 1 football season was not statistically greater in the football population than in noncontact control athletes (P = .55). CONCLUSIONS: Adolescent football players have a greater prevalence of microhemorrhages compared with adolescent athletes who have never engaged in contact sports. While microhemorrhage incidence during 1 season is not significantly greater in adolescent football players compared to adolescent controls, there is a temporal association between playing football and the appearance of new microhemorrhages.

Discriminating between iron deficiency anemia and anemia of chronic disease using traditional indices of iron status vs transferrin receptor concentration.

We compared the ability of soluble serum transferrin receptor (TfR) concentration, quantified using the R&D Systems (Minneapolis, MN) enzyme-linked immunosorbent TfR assay, with other, more traditional indicators of iron status (total iron binding capacity [TIBC], mean corpuscular volume [MCV], percent transferrin saturation [%TS], RBC distribution width [RDW], and serum iron concentration [SIC]) for discriminating between patients with iron deficiency anemia (IDA) or anemia of chronic disease (ACD). The TfR concentration was determined in 72 serum samples selected from men and nonpregnant women classified biochemically on the basis of ferritin concentration as having IDA (n = 41) or ACD (n = 31). By using receiver operating characteristic curve analysis, the diagnostic accuracy of the various indicators of iron status that we evaluated for discriminating between IDA and ACD decreased in the following order: TIBC > TfR > MCV > (%TS = RDW) > SIC. There was no significant difference between the diagnostic accuracy of TIBC and TfR. Thus, the routine measurement of TfR offers no advantage over TIBC for discriminating between people with biochemically defined IDA or ACD.

Soluble transferrin receptor (sTfR) concentration quantified using two sTfR kits: analytical and clinical performance characteristics.

We compared the analytical and clinical performance characteristics of the Ramco and R&D Systems enzyme-linked immunosorbent assays (ELISAs) for quantifying serum levels of soluble transferrin receptor (sTfR). In addition, we determined both the number of samples required to determine the true individual mean sTfR concentration for a single individual and the critical difference (CD) between serial measurements that indicates a statistically significant change in sTfR concentration. sTfR concentration was determined in 127 serum samples selected retrospectively from males (n=32) and non-pregnant (n=40) and pregnant women (n=55). Intra- and inter-assay precision for both methods was good (CV values 5--10%) to excellent (CV values <5%) over a wide range of sTfR concentrations. Correlation between these methods was good (r=0.93); however, sTfR values by the R&D kit were approximately 2.9 times higher than values obtained using the Ramco kit on the same serum samples. Nevertheless, receiver-operator characteristic (ROC) curve analysis demonstrated that the diagnostic accuracy of both assays in discriminating between patients with iron-deficiency anemia (IDA) or anemia of chronic disease (ACD) was high (area-under-the-curve (AUC) values >0.95) and not significantly different (P=0.480). We determined that a minimum of 8 samples are required to determine an individual's true sTfR concentration, while a >40% difference between serial sTfR measurements would be required to indicate a statistically significant change in sTfR concentration. We concluded that both the Ramco and R&D Systems sTfR methods have similar analytical and clinical performance characteristics and were likely to be equally useful in discriminating between patients with biochemically defined IDA or ACD.

Killing of flies in electrocuting insect traps releases bacteria and viruses.

Electrocuting insect traps (EIT) are popular devices frequently used by homeowners and food handlers attempting to localize the control of flying insects, including the ubiquitous house fly (Musca domestica L.). The traps contain a visual attractant and a high-voltage metal grid. Upon contact with the grids, the insects are disintegrated by the high voltage. As part of a systematic evaluation of EITs and their role in infectious disease spread, we quantitated spread of bacteria and a bacterial virus during electrocution of house flies. We loaded flies with Serratia marcescens or with the Escherichia coli phage PhiX174 and placed sprayed or fed flies into a room containing an EIT. While flies were being electrocuted, liberated particles and bacteria were assayed via agar plates or via air filtration samplers. Sprayed flies released one of every 10,000 of the added bacteria or viruses, and fed flies released one of every 1,000,000 of the consumed bacteria or viruses. Results of our studies suggest EITs could play a role in the spread of infectious disease agents, but the potential is influenced by the insect's route of contamination.

Adverse effects of microgravity on the magnetotactic bacterium Magnetospirillum magnetotacticum.

Bacteria that contain magnetosomes display magnetotaxis and align themselves to the earth's magnetic field. When magnetotactic bacteria were first isolated several decades ago it was presumed that geomagnetic orientation allowed magnetotactic bacteria to orient themselves downward towards sediments where the habitat is favorable to their growth and metabolism. As more species of magnetotactic bacteria have been isolated and studied, differences in magnetotactic responses have been observed which suggested that the primary role of magnetosomes might simply be to enhance a microorganism's response to gravity. To resolve if gravity influences magnetotactic behavior in bacteria, Magnetospirillum magnetotacticum was used to examine magnetotaxis in the absence of gravity. Experiments to compare the orientation of bacteria to north- or south-pole magnets were conducted in normal gravity and in the microgravity environments aboard the Space Shuttle and Space Station MIR. In each of the microgravity situations studied, bacteria were impaired in their ability to orient to magnets and the failure to exhibit magnetotaxis appeared to be a function of the loss of magnetosomes. The disappearance of aggregated magnetosomes seemed to correlate with a general loss of cellular integrity in microgravity.

Flies and their bacterial loads in greyhound dog kennels in Kansas.

Breeders of greyhound dogs traditionally feed racing animals and nursing bitches raw meat, and that meat generally is obtained frozen from commercial renderers. Previous studies have shown that the rendered meat is frequently contaminated with enteric bacteria, including Salmonella spp., and that during thawing the rendered meat is exposed to filth flies common in dog kennels. Nursing greyhound pups tend to experience a high morbidity and mortality from intestinal infections, and we attempted to determine in this study whether enterics could be spread to pups through contaminated flies. At intervals during 1995 and 1996, flies were trapped or were net-collected from 10 dog breeding kennels in the region around Abilene, KS. Trapped flies were identified and counted to determine population numbers, and netted flies were cultured in tetrathionate broth and streaked to medium selecting for Salmonella sp. and other lactose-negative Gram (-) bacteria. The relative numbers of different fly species varied with the sampling method, but traps and sweep nets produced similar proportions of the different fly species. Blow flies were twice as likely to be contaminated with enteric bacteria as any other fly. The most common enteric bacteria found were Proteus spp., followed by Providencia spp., Pseudomonas spp., and Salmonella spp. The incidence of Salmonella and Proteus spp. seemed to correlate more with accessibility of flies to dog excrement than to rendered meat. The apparent high incidence of enteric contamination of filth flies clearly implicates them as vectors of enteric diseases in kennels.

Autopsy, radiographic, and prenatal ultrasonographic examination of a stillborn fetus with femoral facial syndrome.

Femoral facial syndrome (FFS) is comprised of cleft palate, micrognathia, short or absent femora, and vertebral and genitourinary malformations. We report on a stillborn fetus with FFS delivered to a mother with gestational diabetes. Prenatal ultrasound examination showed abnormalities at 21 weeks of gestation; prior ultrasound findings were interpreted as normal. Long bone histology showed disorganization of the growth plate with a relative decrease in cartilaginous matrix and vacuolization and binucleation of the chondrocytes.

Methods for increasing survivability during storage of exponentially growing bacteria.

A protocol has been developed for storing Gram (-) bacterial cells at 0 degrees C, which allows greater than 90% of stored cells to retain colony-forming ability for up to 60 days. The protocol, which yields essentially identical results when used with Escherichia coli or Pseudomonas aeruginosa, does not enhance survivability of Bacillus cereus. The greatest and longest survival is enjoyed when exponentially growing cells in minimal-glucose medium are deprived of carbon for about 9 h, supplemented with 750 microg/ml chloramphenicol, and immediately placed at 0 degrees C. By decreasing the period of carbon starvation from 9 to 5 h, or increasing the period of carbon starvation from 9 to 12 h, both the ultimate survival rate and kinetics of loss of culturability are affected. Survival enhancement induced by chloramphenicol is not similarly induced by kanamycin.

Effects of microgravity on the binding of acetylsalicylic acid by Rhizobium leguminosarum bv. trifolii.

Bacteroids can be induced in vitro by treating growing Rhizobium leguminosarum bv. trifolii with succinic acid or succinic acid structural analogs like acetysalicylic acid. Quantitating bacteroid induction by measuring acetylsalicylic binding under normal (1 g) conditions showed two forms of binding to occur. In one form of binding cells immediately bound comparatively high levels of acetylsalicylic acid, but the binding was quickly reversed. The second form of binding increased with time by first order kinetics and reached saturation in 40 s. Similar experiments performed in the microgravity environment aboard the NASA 930 aircraft showed only one form of binding and total acetylsalicyclic acid bound was 32% higher than at 1 g.

Antimicrobial activity of dogwood fruits (Cornus drummondii) from winter food caches of eastern woodrats (Neotoma floridana).

Post-harvest loss of resources to microbial competitors affects a number of species of animals, and many of them have evolved behaviors that reduce the likelihood of such loss. We previously described the inhibition ofBacillus psychrophilus by dogwood fruits (Cornus drummondii), which can comprise a significant portion of the winter food cache of the eastern woodrat (Neotoma floridana). The present study is a further investigation of the ability of dogwood fruits to inhibit growth ofB. psychrophilus. Additionally, we tested the antimicrobial activity of dogwood fruits onEscherichia coli andStaphlococcus aureus. Results of our study indicate that fresh fruits inhibit the growth ofB. psychrophilus andS. aureus but notE. coli. However, fruits taken from woodrat dens after four and six months storage inhibited growth of all three species of bacteria. Further, the strength of inhibition increased over the test period. We suggest the presence of an inhibitor allows woodrats to compete effectively for a valuable and limited food resource.

Rhizobium trifolii 0403 Is Capable of Growth in the Absence of Combined Nitrogen.

Rhizobium trifolii 0403 was treated with 16.6 mM succinate and other nutrients and thereby induced to grow in nitrogen-free medium. The organism grew microaerophilically on either semisolid or liquid medium, fixing atmospheric nitrogen to meet metabolic needs. Nitrogen fixation was measured via N incorporation (18% N enrichment in 1.5 doublings) and acetylene reduction. Nitrogen-fixing cells had a K(m) for acetylene of 0.07 atm (ca. 7.09 kPa), required about 3% oxygen for optimum growth in liquid medium, and showed a maximal specific activity of 5 nmol of acetylene reduced per min per mg of protein at 0.04 atm (ca. 4.05 kPa) of acetylene. The doubling time on N-free liquid medium ranged from 1 to 5 days, depending on oxygen tension, with an optimum temperature for growth of about 30 degrees C. Nodulation of white clover by the cultures showing in vitro nitrogenase activity indicates that at least part of the population maintained identity with wild-type strain 0403.

Stimulation of Nodulation in the Clover-Rhizobium trifolii 0403 System by Penicillin and Mecillinam.

The number of nodules produced per clover seedling inoculated with Rhizobium trifolii 0403 can be increased almost 2-fold by the addition of penicillin or mecillinam. Two-day-old dutch white clover seedlings grown in 250 milliliter boston round jars containing agar-solidified plant growth medium were inoculated with exponentially growing Rhizobium trifolii 0403 cells. Penicillin or mecillinam (100 micrograms per milliliter) were added immediately or after 24 hours. Following 42 days growth, 10 replicate sets of 5 plants for each treatment were assayed for nodule number, plant dry weight, and Kjeldahl nitrogen. Both antibiotics increased nodule number, plant dry weight, and Kjeldahl nitrogen. Increases in nodule number and dry weight were statistically significant. The range of values in Kjeldahl nitrogen was so extensive as to make the data insignificant at the P < 0.05 level, however nodule number, plant dry weight, and Kjeldahl nitrogen displayed a significant correlation with each other. There were no significant differences in treatment with either antibiotic or with time of treatment. Nodule number increased by about 85%, and plant dry weight and nitrogen increased by about 30%.

Response of bacteria isolated from a pristine prairie stream to concentration and source of soluble organic carbon.

Responses of native aquatic bacteria to source and concentration of dissolved organic carbon was observed by using gradient plates. Dissolved organic carbon of prairie (grasses) or gallery forest (bur oak) origin was used as the carbon source in these observations. Bacteria isolated from sediments in the grassland reaches of a prairie stream showed growth only on grass leachates. Bacteria isolated from the gallery forest reaches of the same stream were able to grow on plates made from either source of leachate. The differing quality, quantity, and rate of supply of these dissolved organic carbon sources should select for variation in the bacterial assemblages.

Fine Structure of Succinate-Swollen Rhizobium trifolii 0403.

Transmission electron micrographs of glutaraldehyde- OsO(4)-fixed Rhizobium trifolii 0403 before and after cells were treated with 16.6 mM succinate showed that treated cells increased in mass by increasing cytoplasmic volume. The morphology of succinate-treated cells was identical to that of bacteroids, and the appearances of the envelope and periplasmic space were similar. The primary difference was in inclusion number and type.

Succinate-Induced Morphology of Rhizobium trifolii 0403 Resembles That of Bacteroids in Clover Nodules.

Morphological changes which accompany nutrient enrichment of Rhizobium trifolii 0403 were studied. Assays of cell number and size coupled with scanning electron microscopy and immunofluorescence microscopy showed that succinate induces cells to stop dividing in vitro and to swell either in the cell center or at one cell pole. The extent and frequency of in vitro cell swelling were in direct relation to the concentration of succinate added to the enrichment medium. The in vitro swelling of cells in 16.6 mM succinate plus Casamino Acids, glucose, and yeast extract closely resembled that of bacteroids of R. trifolii 0403 in nitrogen-fixing nodules of white clover. We hypothesize that succinate may be involved in the transformation of vegetative bacteria into the bacteroid morphology found in nitrogen-fixing nodules.

Effect of carbon source on growth rate and phospholipid composition of Escherichia coli 15T- and an unsaturated fatty acid auxotroph.

Escherichia coli 15T- was grown with glucose, succinic acid, aspartic acid, oleic acid, and oleic plus aspartic acids as carbon sources, and a fatty acid auxotroph derived from 15T- was grown on oleic acid and oleic plus aspartic acids. The doubling time, phospholipid composition, phosphorus content, and the fatty acid composition of the phospholipids of cells in each of the media were determined. In all cases, phosphatidylethanolamine was the major phospholipid present; but with 15T- its concentration was inversely proportional to the doubling time in unsupplemented media. With the auxotroph the phosphatidylethanolamine concentration was essentially unchanged with growth. Total lipid phosphorus was inversely proportional to doubling time, an effect particularly evident with the auxotroph. Without oleic acid supplementation, the major effects of carbon source on fatty acid composition are decreases in the content of palmitoleic acid and increases in the content of cis-9, 10-methylene hexadecanoic acid as growth rate decreases. Oleic acid supplementation elevated 18:1 fatty acid content in both 15T- and the auxotroph.

Nondividing, Bacteroid-Like Rhizobium trifolii: In Vitro Induction Via Nutrient Enrichment.

Rhizobium trifolii 0403 maintained in exponential phase via periodic dilution doubled in 210 min in mannitol-salts medium and doubled in 244 min in glycerolsalts. In both media, cell number and optical density increased in parallel. When exponentially growing cells in either medium were supplemented with a mixture of glucose, Casamino Acids, succinate, and yeast extract, optical density continued to increase but within less than the time required for one doubling, division ceased. The increase in optical density coupled with division cessation resulted in the formation of large, pleomorphic, nondividing cells. Large cells apparently increased in size as a result of swelling only at regions of most recent cell envelope synthesis. Greater than 95% of the cells in a population swelled, and commitment to swelling occurred within two doubling time equivalents. Swollen cells eventually reached a characteristic maximum size and exhibited osmotic fragility.

Variations in the cell cycle of slow-growing Escherichia coli 15T-.

DNA synthesis was measured via 3H-thymidine pulses, and patterns of incorporation were compared to cell number increases during synchronous division of Escherichia coli 15T- (555-7) in M9.01% aspartate. The timing of chromosome initiation, duration of chromosome synthesis, and the D period were found to vary over the three cycles of division analysed. In addition, the D period measured by comparing 3H-thymidine incorporation and concomitant division, differed from D period measurements made by timing division cessation in DNA synthesis inhibitors.

Temperature-induced differences in growth rate and fatty acid composition in two strains of Escherichia coli 15T-.

The saturated/unsaturated fatty acid ratio of Escherichia coli 15T- decreases almost threefold as growth temperature decreases from 43 to 27 degrees C, wheras the ratio of a fast-growing mutant derived from 15T- changes only half as much. Strain 15T- experiences a 2.4-fold change in doubling time across this temperature range, but doubling time in the mutant changes 3.3-fold.