Optical scatter imaging detects mitochondrial swelling in living tissue slices

Mitochondrial swelling is observed in neuronal injury and is a key event in many pathways to cell death. Currently, there is no technique for directly measuring mitochondrial size changes within living tissue slices with a field of view of several millimeters. In this paper, we test our hypothesis that Mie light-scatter theory can be used to study mitochondrial swelling in living tissue sections. Using a unique dual-angle scatter ratio (DASR) optical imaging system previously demonstrated to be sensitive to latex particle size changes and N-methyl-D-aspartate (NMDA) treatment of hippocampal slices, we studied mitochondrial swelling induced by 500 microM NMDA treatment of hippocampal slices. We observed a strong (R(2) = 0.73) and significant (P < 0.000005) correlation between the electron microscopy-determined diameters of swollen, intact mitochondria and the DASR imaging. We examined the robustness of the technique by evaluating the correlation between the dual-angle scatter ratio and the diameter of the dendrites, observed to swell, in NMDA-treated slices and found no correlation (R(2) = 0.06). The advantage of DASR imaging over electron microscopy or other methods of studying mitochondrial swelling is the sensitivity of DASR imaging to mitochondrial swelling over a large field of view (>9 mm(2)) in an intact tissue slice. This novel technique may allow for the study of regional changes in mitochondrial swelling and recovery as sequential events within a single specimen. This technique will eventually be useful in studying the efficacy of stroke and other disease therapies targeting mitochondrial swelling.     

Analysis of N-ras gene mutation and p53 gene expression in human hepatocellular carcinomas

AIM:To study the relationship between N-ras gene mutation and p53 gene expression in the carcinogenesis and the development of human hepatocellular carcinomas (HCC).METHODS:The N-ras gene mutation and the p53 gene expression were analyzed in 29 cases of HCC by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and immunohistochemistry.RESULTS:Thirteen cases of HCCs were p53 positive (44.8%), which showed a rather high Cpercen-tage of p53 gene mutation in Guangxi. The aberrations at N-ras codon 2-37 were found in 79.31% of HCCs and 80.77% of adjacent non-tumorous liver tissues. More than 2 point mutations of N-ras gene were observed in 22 cases (75.86%). Twelve cases (41.37%) of HCCs showed both N-ras gene mutation and p53 gene expression.CONCLUSION:N-ras gene and p53 gene may be involved in the carcinogenesis and the development of HCC.That 38% of HCCs with N-ras gene mutation did not express p53 protein indicates that some other genes or factors may participate in the carcinogenesis and the development of HCC.     

Slope analysis of somatosensory evoked potentials in spinal cord injury for detecting contusion injury and focal demyelination

In spinal cord injury (SCI) research there is a need for reliable measures to determine the extent of injury and assess progress due to natural recovery, drug therapy, surgical intervention or rehabilitation. Somatosensory evoked potentials (SEP) can be used to quantitatively examine the functionality of the ascending sensory pathways in the spinal cord. A reduction of more than 50% in peak amplitude or an increase of more than 10% in latency are threshold indicators of injury. However, in the context of injury, SEP peaks are often obscured by noise. We have developed a new technique to investigate the morphology of the SEP waveform, rather than focusing on a small number of peaks. In this study, we compare SEP signals before and after SCI using two rat models: a contusion injury model and a focal experimental autoimmune encephalomyelitis model. Based on mean slope changes over the signal, we were able to effectively differentiate pre-injury and post-injury SEP values with high levels of sensitivity (83.3%) and specificity (79.2%).     

A study of potential vestibulotoxic effects of once daily versus thrice daily administration of tobramycin

Tobramycin 5.1 mg/kg/day was administered for 9 days to 20 healthy human volunteers to determine the potential vestibulotoxicity of once daily versus thrice daily treatment regimens. Vestibular function was tested carefully before, during, and following drug administration, using a battery of postural and caloric tests. Although subjects receiving the once daily regimen performed better on the postural tests, we feel this does not necessarily indicate an adverse effect in either group. The caloric test was the primary measure of vestibular function in this experiment, and this test appeared to demonstrate a slight adverse effect during drug administration of equal magnitude in both treatment groups. The results of this experiment indicate that there is no significant difference in the risk of adverse vestibular effects between once daily and thrice daily tobramycin administration.     

Anterior thalamic mediation of experimental seizures: selective EEG spectral coherence

PURPOSE: Physiological evidence has shown that the anterior thalamus (AN) and its associated efferents/afferents constitute an important propagation pathway for pentylenetetrazol (PTZ)-mediated generalized seizures in rodents. Previous work demonstrated metabolic, physical, chemical, and electrical stimulation data supporting a role for AN in the expression of PTZ seizures. We now extend these observations through examination of neuroelectric signal indicators during seizure epochs. We show that the EEG recorded from AN is highly coherent with surface cortical (CTX) EEG during the immediate preconvulsant period and during the ictal stateough. METHODS: Awake rats were continuously infused with PTZ until clonic seizures were recorded by using both subcortical AN, posterior thalamus (PT), or hippocampal (HPC) bipolar electrodes and cortical EEG. Through the signal-analysis techniques of ordinary and partial coherence, it was possible to focus selectively on signal correlations between AN and CTX (AN/CTX) by removing the effects of unaffiliated regions such as PT and HPC. RESULTS: Coherence of PT/CTX was observed to be modest, and partial coherence of PT/CTX with the effects of AN/CTX removed did not improve the signal coherence of PT/CTX (PT/CTX-AN). In contrast, AN/CTX coherence was observed to be high, with undiminished correlation when PT/CTX influence was removed (AN/CTX-PT). The most robust band of AN/CTX coherence was centered around the spike-wave clonic frequency of 1-3 Hz. Partial multiple coherence-analysis techniques were used to remove the possible signal contributions from hippocampus in addition to PT. The AN/CTX coherence remained fully preserved in the low-frequency bands. CONCLUSIONS: These data provide electrophysiologic evidence supporting the special role of the anterior thalamus in the propagation of seizure activity between subcortex and cortex.     

Presyncope caused by central hypovolaemia is not preceded by evoked potential alterations.

The mechanism(s) responsible for the onset of presyncope during a central hypovolaemic challenge have gone undefined for many years. It has been speculated that a decrease in cerebral blood flow initiates presyncopal responses, which in turn lead to greater decreases in cerebral oxygen delivery and unconsciousness. Somatosensory evoked potentials (SEP) were monitored as a measure of cerebral functioning in ten subjects during presyncopal symptom limiting lower body negative pressure (a central hypovolaemic challenge). SEP latency and amplitudes have been correlated with cerebral oxygen uptake, so SEP activity can serve as an indirect indicator of cerebral homeostasis. SEPs were generated by electrically stimulating the median nerve and recoding the resulting potentials over the contralateral cerebral cortex. While heart rate and mean blood pressure both fell at presyncope, there were no changes noted in either SEP latency or amplitude at any point before (latency = 22.9 +/- 9 ms; amplitude = 2.86 +/- 0.24 microV), during (22.6 +/- 0.9 ms; 2.68 +/- 0.2 microV), or after (22.7 +/- 0.9 ms; 2.37 +/- 0.23 microV) the occurrence of presyncope. We conclude that the onset of presyncope is not associated with a decrease in cerebral function.     

MEMSurgery: an integrated test-bed for vascular surgery

Many surgical procedures require skillful manipulations of blood vessels, especially in conventional invasive or minimally invasive surgical procedures. Current surgical methods do not allow the surgeon to receive any real time feedback of the tissue properties when operating on the vessel. As a result, the unintentional application of excessive force may damage the blood vessel. To minimize such trauma, and to study the interaction of surgical instruments with the vessel structure, we have developed an integrated surgical testbed called MEMSurgery (Microelectromechanical Sensory augmented Surgery). The test-bed integrates four elements: a) force sensors mounted on surgical appliances, b) a feedback control mechanism utilizing the intrinsic mechanical properties of the blood vessel, c) feedback of the force applied on the tissue back to the surgeon through a haptic feedback device, and d) visual feedback by a graphical computer model of the vessel. Finally, we evaluate the performance of MEMSurgery by testing the hypothesis that the combination of haptic feedback, feedback control based on vascular mechanical properties, and real-time visual representation of the vessel will help the surgeon decrease the probability of applying excess force while occluding the blood vessel. To this end, we designed a rodent experimental model to obtain the ideal minimum occlusion force (MOF). After a series of human performance studies, and subsequent comparison to direct application of force on the forceps (without feedback), the results show that the probability of applying reasonable MOF increases from 35.5% to 80%. After a brief training period, the probability increases to 90%.     

Pharmacokinetics of enfuvirtide in pediatric human immunodeficiency virus 1-infected patients receiving combination therapy

BACKGROUND: Enfuvirtide is the first of a new class of antiretroviral agents, the fusion inhibitors. OBJECTIVES: The primary objective of this analysis was to evaluate the pharmacokinetics of 2.0 mg/kg enfuvirtide in human immunodeficiency virus 1 (HIV-1)-infected children and adolescents when administered in combination with at least 3 other antiretrovirals. METHODS: Twenty-five HIV-1-infected pediatric patients (5-16 years of age) enrolled in an ongoing phase I/II study were included in this analysis. Patients received enfuvirtide 2.0 mg/kg sc twice daily (bid) for at least 7 days. Blood samples were collected on day 7, and plasma concentrations of enfuvirtide and its metabolite were measured by a validated liquid chromatography-tandem mass spectrometry method. Pharmacokinetics measures [Cmax, tmax, Ctrough, and area under the concentration time curve time 0 to 12 hours (AUC12 hours)] were calculated from plasma concentration-time data by standard noncompartmental methods. RESULTS: There was no significant difference between children and adolescents for enfuvirtide Cmax (6.43 versus 5.88 microg/mL), Ctrough (2.87 versus 2.98 microg/mL) and AUC12 hours (56.1 versus 52.7 hours . microg/mL). Similarly no significant differences were found when the pharmacokinetic measures were compared based on sexual maturity stages. A post hoc regression analysis based on AUC12 hours showed that body weight-adjusted dosing of enfuvirtide provides drug exposure that is independent of age group, body weight and body surface area. CONCLUSIONS: Body weight-adjusted dosing of enfuvirtide, at a dose of 2.0 mg/kg sc bid, in HIV-1-infected pediatric patients at least 5 years of age, provides drug exposure comparable with that previously observed in HIV-1-infected adults after 90 mg sc bid dosing. Drug exposure in children and adolescents is independent of age group, body weight, body surface area and sexual maturity stage.     

Excision of a recurrent mediastinal cyst through mid sternotomy with high thoracic epidural anesthesia as the sole anesthetic—A case report

Conclusion Our case report highlights the possibility of tackling mediastinal, pericardial and pleural problems with regional anesthesia alone even if the surgery requires opening of the pleura.