Redefining the boundaries of the hippocampal CA2 subfield in the mouse using gene expression and 3-dimensional reconstruction

The morphology of neurons in the main divisions of the hippocampal complex allow the easy identification of granule cells in the dentate gyrus and pyramidal cells in the CA1 and CA3 regions of Ammon's horn. However, neurons in the CA2 subfield have been much more difficult to reliably identify. We have recently identified a set of genes whose expression is restricted to either the dentate gyrus, CA1, CA2, or CA3. Here we show that these genes have an essentially nonoverlapping distribution throughout the entire septotemporal extent of the hippocampus. 3-Dimensional reconstruction of serial sections processed for in situ hybridization of mannosidase 1, alpha (CA1), bcl-2-related ovarian killer protein (CA3), and Purkinje cell protein 4 (dentate gyrus+CA2) was used to define the boundaries of each subregion throughout the entire hippocampus. The boundaries observed for these three genes are recapitulated across a much larger set of genes similarly enriched in specific hippocampal subregions. The extent of CA2 defined on the basis of gene expression is somewhat larger than that previously described on the basis of structural anatomical criteria, particularly at the rostral pole of the hippocampus. These results indicate that, at least at the molecular level, there are robust, consistent genetic boundaries between hippocampal subregions CA1, CA2, CA3, and the dentate gyrus, allowing a redefinition of their boundaries in order to facilitate functional studies of different neuronal subtypes in the hippocampus.     

Inhibitory activities of colicins against Escherichia coli strains responsible for postweaning diarrhea and edema disease in swine

The efficacies of colicins E1 and N against Escherichia coli strains responsible for postweaning diarrhea and edema disease, two of the most prevalent disease problems for pigs in the United States, were determined in vitro. These proteins may provide an environmentally sound means for the prevention of these infections in swine.     

Small molecule activators of the heat shock response and neuroprotection from stroke

Various cellular defense pathways are mobilized in response to stress that serve to limit potential damage to organelles and biochemical pathways that would disrupt normal cellular function or trigger cell death. Strategies utilized by cells subjected to various forms of stress include the activation of detoxification systems that act to eliminate the primary damaging molecules, remove damaged cellular macromolecules, or restore organelle and macromolecule function in cases where loss of activity is generated by reversible modifications or alterations in conformation (ie, misfolding). Central to many intracellular defense mechanisms that operate to limit damage to protein function are molecular chaperones of the heat shock protein (HSP) family. This review briefly discusses the molecular mechanisms that are thought to dictate the well-established neuroprotective effects of HSPs and highlight the recent attempts to use pharmacologic approaches to activate this important cellular defense pathway.     

Examination of heat stress and stage of lactation (early versus late) on fecal shedding of E. coli O157:H7 and Salmonella in dairy cattle

Mature, healthy lactating dairy cattle were sampled on two farms in the southwestern United States to examine the effects of heat stress (Experiment I) and stage of lactation (Experiment II) on the fecal shedding of E. coli O157:H7 and Salmonella. To examine the effects of heat stress, fecal samples were collected from 45 cows at 7:00 AM (coolest part of the day) and 5:00 PM (hottest part of the day) in August 2002 on a 250 cow dairy. The study was replicated one month later (n = 170 total samples). A temperature-heat index (THI) was calculated for each sampling time. In Experiment II, stage of lactation was examined by sampling lactating dairy cattle early [< 60 days in milk (DIM)] and late (> 150 DIM) in the lactation cycle in the summer of 2001. The study was replicated the following summer (60 cows/group/replicate; n = 240 total samples). For Experiment I, THI averaged 75 and 82 for the AM and PM samplings, respectively, indicating the cows were beginning to experience heat stress in the morning and by afternoon were in severe heat stress. The shedding of E. coli O157:H7 tended to be higher (p = 0.09) in the afternoon sampling of the first replicate, however was not different in the second replicate or when both replicates were pooled (p > 0.10). Salmonella shedding was not different (p > 0.10) at any sampling time with nearly 100% of the cows positive. Stage of lactation had no effect on the number of cows shedding E. coli O157:H7 (p > 0.10). Salmonella shedding tended to be higher (p = 0.09) in early lactation cows in the first replicate, while in the second replicate more late lactation cows were shedding Salmonella (p < 0.05); however, there were no differences due to stage of lactation when replicates were pooled (p > 0.10). While further research is needed, results of this research highlight the variability in pathogen shedding in healthy dairy cattle and indicate that environmental factors and/or production demands may influence shedding patterns of E. coli O157:H7 and Salmonella.     

Botulinum toxin type B: an overview of its biochemistry and preclinical pharmacology

Produced by Clostridium botulinum, botulinum toxins are high molecular weight protein complexes consisting of the neurotoxin and additional nontoxic proteins that function to protect the toxin molecule. The neurotoxin acts to inhibit the release of acetylcholine at the neuromuscular junction, causing muscle paralysis. Purified toxin complexes have found a niche in the treatment of clinical disorders involving muscle hyperactivity. The different serotypes are structurally and functionally similar; however, specific differences in neuronal acceptor binding sites, intracellular enzymatic sites, and species sensitivities suggest that each serotype is its own unique pharmacologic entity. Recently, botulinum toxin type B has been developed as a liquid formulation to avoid the lyophilization (vacuum-drying) and reconstitution processes associated with decreasing the potency and stability of current type A toxin preparations. Biochemical tests were conducted to evaluate the quality of toxin in this formulation. In 3 consecutive manufacturing lots, the botulinum toxin type B complex was found to be highly purified, intact, uniform, and consistent from lot to lot. Also, it showed long-term stability at refrigerator and room temperatures (2 to 25 degrees C). Electrophysiologic studies in cynomolgus monkeys showed that botulinum toxin type B is effective in paralyzing injected muscle groups, with minimal spread to relatively distant noninjected muscles.     

Chest radiograph interpretation by medical students

AIM: To assess the ability of final year medical students to interpret conventional chest radiographs. MATERIALS AND METHODS: Ten conventional chest radiographs were selected from a teaching hospital radiology department library that were good radiological examples of common conditions. All were conditions that a medical student should be expected to recognize by the end of their training. One normal radiograph was included. The radiographs were shown to 52 final year medical students who were asked to describe their findings. RESULTS: The median score achieved was 12.5 out of 20 (range 6-18). There was no difference between the median scores of male and female students (12.5 and 12.3, respectively, p=0.82) but male students were more likely to be certain of their answers than female students (median certainty scores 23.0 and 14.0, respectively). The overall degree of certainty was low. On no radiograph were more than 25% of students definite about their answer. Students had received little formal radiology teaching (2-42 h, median 21) and few expressed an interest in radiology as a career. Only two (3.8%) students thought they were good at interpreting chest radiographs, 17 (32.7%) thought they were bad or awful. CONCLUSION: Medical students reaching the end of their training do not perform well at interpreting simple chest radiographs. They lack confidence and have received little formal radiological tuition. Perhaps as a result, few are interested in radiology as a career, which is a matter for concern in view of the current shortage of radiologists in the UK.     

Dynamic nature of electrocardiographic waveform predicts rescue shock outcome in porcine ventricular fibrillation

STUDY OBJECTIVE: Survival decreases with duration of ventricular fibrillation, and it is possible that failed rescue shocks increase myocardial damage. Structure in the ECG signal during ventricular fibrillation can be quantified by using the scaling exponent, a dimensionless measure that correlates with ventricular fibrillation duration. This study examined whether the scaling exponent could predict rescue shock success and whether unsuccessful rescue shocks altered the structure of the ventricular fibrillation waveform and the responsiveness to subsequent rescue shocks. METHODS: Ventricular fibrillation was electrically induced in 44 anesthetized swine, which were randomly assigned to receive 70-J biphasic rescue shocks at 2, 4, 6, 8, or 10 minutes. If rescue shocks failed, up to 2 subsequent rescue shocks were performed at 2-minute intervals. The scaling exponent was calculated at 1-second intervals from ECG to quantify the organization of the ventricular fibrillation waveform. RESULTS: A total of 92 rescue shocks were delivered, of which 23 successfully converted ventricular fibrillation to an organized rhythm (immediate success). After these 23 rescue shocks, 14 swine sustained organized rhythms for more than 30 seconds (sustained success). Lower scaling exponent values were associated with increased probability of successful rescue shocks. Receiver operating characteristic curves had an area under the curve of 0.86 for immediate rescue shock success and 0.93 for sustained rescue shock success. Failed rescue shocks increased the rate of scaling exponent increase over time but did not appear to affect subsequent rescue shock success when the scaling exponent was taken into account. CONCLUSION: Highly deterministic ventricular fibrillation, reflected by a low scaling exponent, predicted rescue shock success regardless of antecedent failed rescue shocks. In addition, unsuccessful rescue shocks might decrease post-rescue shock ventricular fibrillation waveform organization.