Growth factor signaling, mediated via receptor tyrosine kinases (RTKs), needs to be tightly regulated in many developmental systems to ensure a physiologically appropriate biological outcome. At one level this regulation may involve spatially and temporally ordered patterns of expression of specific RTK signaling antagonists, such as Sef (similar expression to fgfs). Growth factors, notably FGFs, play important roles in development of the vertebrate ocular lens. FGF induces lens cell proliferation and differentiation at progressively higher concentrations and there is compelling evidence that a gradient of FGF signaling in the eye determines lens polarity and growth patterns. We have recently identified the presence of Sef in the lens, with strongest expression in the epithelial cells. Given the important role for FGFs in lens developmental biology, we employed transgenic mouse strategies to determine if Sef could be involved in regulating lens cell behaviour. Over-expressing Sef specifically in the lens of transgenic mice led to impaired lens and eye development that resulted in microphthalmia. Sef inhibited primary lens fiber cell elongation and differentiation, as well as increased apoptosis, consistent with a block in FGFR-mediated signaling during lens morphogenesis. These results are consistent with growth factor antagonists, such as Sef, being important negative regulators of growth factor signaling. Moreover, the lens provides a useful paradigm as to how opposing gradients of a growth factor and its antagonist could work together to determine and stabilise tissue patterning during development and growth.

We have compared spatial patterns of somatosensory evoked magnetic fields (SEFs) to stimulation of the ulnar and median nerves at the wrist. An oddball paradigm was used additionally to examine whether an infrequent change in the stimulation site would alter the field pattern. The response consisted of 3 parts: an early small deflection at 22-28 msec, a large deflection peaking between 34 and 86 msec, and a late deflection at 110-180 msec. The wave forms and amplitudes of the responses to ulnar and median nerve stimulation were similar, without any additional deflections for the infrequent stimuli. The field patterns, which were interpreted in terms of the dipole model, could be explained by activation of the primary sensorimotor cortex during all peaks of the response. For the early parts of the response at 22-46 msec, the locations of the equivalent sources for median and ulnar nerve stimulation differed from each other, in agreement with the known somatotopy of SI. No somatotopical order was found for the sources of the later deflections.

Functional magnetic resonance imaging (fMRI) was used to investigate neural changes as a function of category learning in normals (n=8). Subjects were trained to classify patterns of dots into four categories over 4 consecutive days. fMRI monitored the changes that occurred during learning prior to training and then following each training session. During fMRI, subjects determined whether two patterns of dots were members of the same category. The behavioral changes that occurred as a result of the training were observed as increases in response accuracy within shortened response times. fMRI illustrated initial increases in volumes of activation distributed across the known visuospatial processing networks. The regions affected by learning were identified as those involved in the planning and execution of eye movements (frontal and supplementary eye fields, FEF and SEF), spatial attention (superior and inferior parietal lobules, SPL and IPL) and visual processing (primary, secondary, and tertiary visual cortices). The volumes of activation then decreased as training progressed further. Of the two proposed mechanisms for learning, that of strengthened connectivity on a given network and that of selection of different networks, our data supports the former.

OBJECTIVE

To investigate the effect of interstimulus interval (ISI) on attentional enhancement of cortical activity in human somatosensory cortices.

METHODS

Somatosensory magnetic fields (SEFs) evoked by electrical stimulation of the right median nerve at ISIs of 0.5, 1, 3 and 5 s were recorded in two separate conditions by using whole head type magnetoencephalography. In the attend-condition: subjects directed attention to the stimuli by watching the stimulated body site and mentally counting the stimuli. In the ignore-condition: subjects ignored the stimuli by reading a book.

RESULTS

Attention to the tactile stimulation amplified SEFs in contralateral secondary (SIT) at ISIs of 1, 3 and 5 s and in ipsilateral SII at ISI of 5 s. There was no effect of attention on the SEF in primary. The ratios of the equivalent current dipole moments calculated for the attend-condition compared to ignore-condition were 1.49 +/- 0.14 at ISI of 1 s, 1.47 +/- 0.29 at 3 s and 1.34 +/- 0.23 at 5 s in the contralateral SII and 1.40 +/- 0.24 at 5 s in the ipsilateral SII.

CONCLUSIONS

Time longer than 0.5 s might be required for active attention genesis and, after it was generated, the cortical activation in the attend condition was proportional to the activity in the ignore condition.

We have studied the maturation of electrocortical (ECoG) activity in fetal sheep and the impact of chronic hypoxemia using a growth restriction model with placental embolizations. Twenty chronically catheterized fetal sheep (control, n=9; hypoxemic, n=11) were monitored at 116-119, 121-126 and 128-134 days gestational age (term=145 days), with ECoG activity scored using automated analysis of amplitude and frequency components to distinguish low-voltage/high frequency (LV/HF) and high-voltage/low frequency (HV/LF) state epochs, along with indeterminate voltage/frequency (IV/F) and transition period activities. We have shown that multiple aspects of ECoG state activity in the ovine fetus undergo maturational change as electrophysiologic measures of brain development. With chronic fetal hypoxemia, some maturational changes continue to occur, i.e. ECoG activity amplitude and 95% SEF, indicating the resiliency of these parameters to adverse conditioning. However, some maturational changes were altered, i.e. LV/HF and HV/LF incidence and duration, and likely regulated and adaptive with a decrease in the brain's nonessential energy needs, while some were altered, i.e. IV/F incidence and duration, and state transition times, and likely indicating a degree of aberrant development in associated control circuitries. This may then have consequences for disturbed sleep-wake patterns during later life and for adverse neurologic sequelae known to be increased in humans born with growth restriction.

In order to test retention of teaching skills acquired on a two-day Teaching Improvement Project course 94 participants from 12 courses were pre-tested using a sef-evaluation questionnaire and then re-tested with the same questionnaire, between one and two years after course attendance. With a 77% post-course response rate significant differences were measured in participants' self-evaluation of their teaching skills, particularly in the use of learning objectives, structuring learning, questioning techniques and self-confidence. A control, 'demonstrating practical procedures', which was not part of the course, showed no significant difference. Many free comments indicated that teachers had used their new teaching skills with students, that they were more confident as teachers and that they felt their students had benefited from their teacher training. A smaller number of comments suggested some teachers had become more reflective.

Transcranial magnetic stimulation (TMS) was applied over the frontal eye field (FEF) and the supplementary eye field (SEF) regions before execution of the first saccade in a double step task. When applied over the FEF, stimulation increased the percentage error in amplitude of the contralateral second saccade as compared to no stimulation. This was due to an interference with retinotopic but not craniotopic gain calculation. Stimulation of the SEF region interfered with saccade ordering. Thus, FEF might participate in target memorization whereas SEF is confirmed to code order information for sequential saccades even in this paradigm of only two consecutive movements.

Development of the CTF MEG system has been advanced with the introduction of a computer processing cluster between the data acquisition electronics and the host computer. The advent of fast processors, memory, and network interfaces has made this innovation feasible for large data streams at high sampling rates. We have implemented tasks including anti-alias filter, sample rate decimation, higher gradient balancing, crosstalk correction, and optional filters with a cluster consisting of 4 dual Intel Xeon processors operating on up to 275 channel MEG systems at 12 kHz sample rate. The architecture is expandable with additional processors to implement advanced processing tasks which may include e.g., continuous head localization/motion correction, optional display filters, coherence calculations, or real time synthetic channels (via beamformer). We also describe an electronics configuration upgrade to provide operator console access to the peripheral interface features such as analog signal and trigger I/O. This allows remote location of the acoustically noisy electronics cabinet and fitting of the cabinet with doors for improved EMI shielding. Finally, we present the latest performance results available for the CTF 275 channel MEG system including an unshielded SEF (median nerve electrical stimulation) measurement enhanced by application of an adaptive beamformer technique (SAM) which allows recognition of the nominal 20-ms response in the unaveraged signal.

BACKGROUND

We studied the effect of variable doses of ketamine on the endpoints of hypnosis, e.g., unresponsiveness to verbal commands (UVC), loss of eyelash reflex (LER), and inhibition of body movement response with or without sneezing to nasal membrane stimulation (INBMR), and processed EEG variables, e.g., bispectral index (BIS), 95% spectral edge frequency (SEF) and median frequency (MF) during propofol infusion.

METHODS

Forty-eight patients received either propofol infusion, 30 mg.kg-1.h-1 (Group P; n = 12) or ketamine bolus, 0.25, 0.5 or 0.75 mg i.v., followed by propofol infusion, 30 mg.kg-1.h-1 + variable dose ketamine infusion, 0.25, 0.5 or 0.75 mg.kg-1.h-1 (Groups PK0.25, PK0.5 and PK0.75; n = 12 each) until UVC, LER and INBMR. BIS, 95% SEF and MF values were monitored and recorded at the endpoints of hypnosis. Propofol and ketamine concentrations were measured at INBMR.

RESULTS

Propofol infusion, 30 mg.kg-1.h-1, induced UVC, LER and INBMR at BIS: 65 +/- 2, 63 +/- 9 and 33 +/- 7; 95% SEF: 17 +/- 3, 17 +/- 4 and 14 +/- 3; and MF values of 5 +/- 2, 5 +/- 3 and 3 +/- 2, respectively. With adjunctive ketamine (Groups PK0.5 and PK0.75), the hypnotic endpoints were achieved at higher BIS and 95% SEF values and lower propofol doses and concentrations as compared to Groups P and PK0.25 (9.9 +/- 5.8 and 9.4 +/- 3.4 vs. 13.4 +/- 4.5 and 14 +/- 5.8 micrograms.ml-1).

CONCLUSIONS

Our results suggest additive interaction between propofol and ketamine (Groups PK0.5 and PK0.75) for achieving the hypnotic endpoints; however, ketamine did not depress the EEG variables in proportion to its hypnotic effect. The paradoxically higher BIS and 95% SEF values at the hypnotic endpoints may be due to lower propofol concentrations and/or no effect of ketamine on the EEG variables.

Spectral edge frequency (SEF) of the EEG was studied in 51 healthy neonates to assess its value as a measure of cerebral maturation. SEF was calculated as the frequency below which 95% of the power in the power spectrum resides. Summated SEF (sSEF) of 4 EEG channels correlated significantly with gestational age (r = 0.83). sSEF varied with behavioural state and with EEG derivation. Intra- and interindividual variability were both greater in preterm compared to term babies. We conclude that SEF is a useful measure of cerebral maturation in the newborn and may have a role in studying stressed newborns.

The cerebellar dorsal vermis lobules VI-VII (oculomotor vermis) and its output region (caudal fastigial nuclei, cFN) are involved in tracking eye movements consisting of both smooth-pursuit and saccades, yet, the exact role of these regions in the control of tracking eye movements is still unclear. We compared the neuronal discharge of these cerebellar regions using a memory-based, smooth-pursuit task that distinguishes discharge related to movement preparation and execution from the discharge related to the processing of visual motion signals or their memory. Monkeys were required to pursue (i.e., go), or not pursue (i.e., no-go) in a cued direction, based on the memory of visual motion direction and go/no-go instructions. Most (>60 %) of task-related vermal Purkinje cells (P-cells) and cFN neurons discharged specifically during the memory period following no-go instructions; their discharge was correlated with memory of no-go instructions but was unrelated to eye movements per se during the action period of go trials. The latencies of no-go discharge of vermal P-cells and cFN neurons were similar, but were significantly longer than those of supplementary eye field (SEF) no-go neurons during an identical task. Movement-preparation signals were found in ~30 % of smooth-pursuit-related neurons in these cerebellar regions and some of them also carried visual memory signals. Our results suggest that no-go neurons are a newly revealed class of neurons, detected using the memory-based pursuit task, in the oculomotor vermis-cFN pathway and that this pathway contributes specifically to planning requiring the working memory of no-go instructions and preparation of tracking eye movements.

Budded virions of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) can enter a variety of non-host cells. The capacity of GP64, AcMNPV's endogenous envelope fusion protein, and SeF, the fusion protein from a gp64(-) baculovirus, to mediate baculovirus entry into vertebrate cells was examined by comparing the transduction efficiencies of engineered AcMNPV variants with either of the two envelope proteins into 17 vertebrate cell lines. At an m.o.i. of 500, GP64-expressing viruses transduced all cell lines with varying efficiencies. Transduction efficiencies of SeF-pseudotyped gp64-null AcMNPV into all cell lines were lower than those of GP64-expressing viruses, and were undetectable in seven cell lines. At an m.o.i. of 50, transduction of all mammalian cell lines transducible by the SeF-pseudotyped gp64-null AcMNPV at an m.o.i. of 500 was no longer detectable. An amplifiable SeF-pseudotyped gp64-null AcMNPV vector with greatly reduced tropism for vertebrate cells may have applications in engineering AcMNPV for targeted transduction.

OBJECTIVE

To describe a clinical encounter (Bedside Examination Exercise [BEE]) used for assessment and teaching in the Massachusetts General Hospital neurology clerkship; to compare results of the BEE with the Harvard Medical School Subjective Evaluation Form (SEF) and National Board of Medical Examiners Shelf examination (Shelf); and to develop a grading system that assesses multiple skills and reflects proficiency.

METHODS

The BEE was administered to 409 students. Final grades were compared with those of 71 students evaluated with the SEF alone. We compared results on the SEF, BEE, and Shelf examination in another 132 students. A composite score was developed, weighted SEF 70%, BEE 15%, and Shelf 15%, to derive the final grade.

RESULTS

The BEE helped limit grade inflation, but did not predict final grade determined by the SEF. Grades from the three test instruments had normal distributions, but different means and SDs: SEF 84% +/- 10.3%; BEE 83% +/- 9.3%; Shelf 69% +/- 8.4%. There was poor agreement among individual students between different tests, even within core competencies. The 70-15-15 composite score had a normal distribution, 81% +/- 8.5%.

CONCLUSIONS

The Bedside Examination Exercise (BEE) is useful for assessing and teaching clinical skills. No single test instrument predicts results of another with acceptable accuracy. Use of complimentary assessment tools (BEE, Subjective Evaluation Form, and Shelf) lessens uncertainty in deriving the composite score, and facilitates evaluation of different attributes. The composite score enables a five-tier grading system that recognizes proficiency, rewards excellence, and provides meaningful feedback. This approach could be generalized to other clerkships.

Thirty-three patients with angina (31 men and 2 women, age 33 to 68 years, 52), as well as signs and symptoms of severe left ventricular dysfunction, were evaluated for coronary revascularization surgery. All had multiple vessel coronary artery disease and at least one prior myocardial infarction. Cardiac catheterization demonstrated abnormally elevated left ventricular end-diastolic pressure (LUEDP), low cardiac output, and depressed resting biplane systolic ejection fraction (SEF) ranging from 18 to 45 per cent (31 per cent). To evaluate potential myocardial function, a premature ventricular contraction was introduced during the ventriculogram and the SEF of the postextrasystolic potentiated (PESP) beat calculated and compared to a sinus beat SEF. Patients were separated into two groups based on the increase in SEF: those with greater than 0.10 augmentation (24 patients) and those with less than 0.10 augmentation (9 patients). Coronary revascularization was carried out with at least two bypass grafts in each patient. The operative mortality in those with more than 0.1 SEF augmentation was 9 per cent (2/24), late mortality rate 5 per cent (1/22), and 20/21 became Class I or II in the follow-up period of 11 to 57 months (25). Operative mortality in those with SEF augmentation of less than 0.1 3/9 33 per cent), late mortality rate 1/6, and only 1/5 achieved Class 1 status during the follow-up period of 10 to 35 months (22) postoperatively. These data suggest that significant augmentation of SEF by a premature ventricular contraction is a simple and useful indicator to aid in selection of patients with left ventricular dysfunction for coronary revascularization.

In tetrapod limbs, an anteriormost digit has common traits of small, short and less-phalange morphology. In this study, we focused on three genes, Mkp3, Sef and Tsukushi (TSK), which have anterior-specific or anterior-prominent expression patterns in the developing limb bud at the autopod-forming stage. The anterior expression is not fixed in the period of limb development, but the expression domains of Mkp3, Sef and TSK change considerably from the distal domain to the anterior domain. This change in expression domains, anterior shift, of these genes involves maintenance of gene expression in the anterior side and downregulation in the posterior side. Manipulated overdose of fibroblast growth factor (FGF) in the presumptive digit 2 region of chick forelimb bud results in elongation of cartilage elements of digit 2, suggesting that attenuated FGF signaling, which Mkp3, Sef, and TSK negatively regulate, provides digit 2-specific traits of morphology. The anterior expression of Mkp3 and Sef but not TSK is conserved also in limb buds of the mouse and gecko, and the anterior shift of these genes, accumulation of their transcripts in the anterior side and appropriate regulation of strength of FGF signaling may control species-specific morphology of the anteriormost digit.

Classic identity negative priming (NP) refers to the finding that when an object is ignored, subsequent naming responses to it are slower than when it has not been previously ignored (Tipper, S.P., 1985. The negative priming effect: inhibitory priming by ignored objects. Q. J. Exp. Psychol. 37A, 571-590). It is unclear whether this phenomenon arises due to the involvement of abstract semantic representations that the ignored object accesses automatically. Contemporary connectionist models propose a key role for the anterior temporal cortex in the representation of abstract semantic knowledge (e.g., McClelland, J.L., Rogers, T.T., 2003. The parallel distributed processing approach to semantic cognition. Nat. Rev. Neurosci. 4, 310-322), suggesting that this region should be involved during performance of the classic identity NP task if it involves semantic access. Using high-field (4 T) event-related functional magnetic resonance imaging, we observed increased BOLD responses in the left anterolateral temporal cortex including the temporal pole that was directly related to the magnitude of each individual's NP effect, supporting a semantic locus. Additional signal increases were observed in the supplementary eye fields (SEF) and left inferior parietal lobule (IPL).

We have studied 46 patients to compare the efficacy of the auditory evoked potential (AEP) index, bispectral index (BIS), 95% spectral edge frequency (SEF) and median frequency (MF) in predicting movement in response to insertion of the laryngeal mask airway (LMA). Anaesthesia was induced with target-controlled infusions of propofol and alfentanil. After loss of eyelash reflex and adequate jaw relaxation, the LMA was inserted without the assistance of a laryngoscope or neuromuscular blocker. Patients who showed any visible spontaneous muscle movement within 1 min of LMA insertion were defined as movers. Values in movers and non-movers at 30 s before LMA insertion were analysed. Only AEP index discriminated between movers and non-movers with a prediction probability of 0.872. BIS, SEF and MF could not predict movement at LMA insertion. AEP index was the most reliable predictor of movement in response to LMA insertion.

Browsing and grazing ruminants are thought to differ in the degree their rumen contents are stratified-which may be due to different characteristics of their respective forages, to particular adaptations of the animals, or both. However, this stratification is difficult to measure in live animals. The papillation of the rumen has been suggested as an anatomical proxy for stratification-with even papillation indicating homogenous contents, and uneven papillation (with few and small dorsal and ventral papillae, and prominent papillae in the atrium ruminis) stratified contents. Using the surface enlargement factor (SEF, indicating how basal mucosa surface is increased by papillae) of over 55 ruminant species, we demonstrate that differences between the SEF(dorsal) or SEF(ventral) and the SEF(atrium) are significantly related to the percentage of grass in the natural diet. The more a species is adapted to grass, the more distinct this difference, with extreme grazers having unpapillated dorsal and ventral mucosa. The relative SEF(dorsal) as anatomical proxy for stratification, and the difference in particle and fluid retention in the rumen as physiological proxy for stratification, are highly correlated in species (n = 9) for which both kind of data are available. The results support the concept that the stratification of rumen contents varies among ruminants, with more homogenous contents in the more browsing and more stratified contents in the more grazing species.

OBJECTIVE

To investigate the effect of conditioning painful stimulation on the early somatosensory magnetic fields (SEF) of test stimulation, in order to clarify the location of the gating effect of pain on tactile response.

METHODS

We used a conditioning stimulus (CS) and test stimulus (TS) paradigm. The CS was applied at the left index finger followed by the TS at the left median nerve. The interstimulus interval between the CS and TS was varied from 100 to 1000 ms. There were two sessions corresponding to two intensities of the CS, painful CS (PCS) and non-painful CS (NPCS). Early components of SEF recorded 20 (1M) and 30 ms (2M) following the TS and the components obtained 20 (1m) and 30 ms (2m) following the CS were analyzed. Each value was compared between the two sessions.

RESULTS

PCS and NPCS attenuated the response of the 2M but not the 1M. The effect of PCS was significantly stronger and lasted longer than that of NPCS. The 1m and 2m components did not differ between PCS and NPCS in terms of amplitude and latency.

CONCLUSIONS

Our data indicated that the early components of the median nerve SEF were affected by a preceding painful stimulation much more than a non-painful stimulus given on the median nerve, and that the sensory gating effect of a painful stimulation on tactile sensation lasted longer than that of a non-painful stimulation. Furthermore, our findings suggested the existence of a 'touch gate' (effect of pain on tactile sensation) at the level of the thalamus or primary somatosensory cortex (SI).

CONCLUSIONS

The finding suggested that the touch gate might lie in the thalamus or SI.

OBJECTIVE

To record the electroencephalographic changes during castration in ponies anaesthetized with halothane and given intravenous (IV) lidocaine by infusion. The hypothesis tested was that in ponies, IV lidocaine is antinociceptive and would therefore obtund EEG changes during castration.

METHODS

Ten Welsh mountain ponies referred to the Department of Clinical Veterinary Medicine, Cambridge for castration under general anaesthesia.

METHODS

Following pre-anaesthetic medication with intramuscular acepromazine (0.02 mg kg(-1)) anaesthesia was induced with IV guaiphenesin (60 mg kg(-1)) and thiopental (9 mg kg(-1)) and maintained with halothane at an end-tidal concentration (FE'HAL) of 1.2%. A constant rate infusion of IV lidocaine (100 microg kg(-1) minute(-1)) was administered throughout anaesthesia. The electroencephalogram (EEG) was recorded continuously using subcutaneous needle electrodes. All animals were castrated using a closed technique. The raw EEG signal was analysed after completion of each investigation, and the mean values of EEG variables (median frequency, spectral edge frequency, total amplitude) recorded during a baseline period (before surgery began) and the removal of each testicle were compared using anova for repeated measures.

RESULTS

Spectral edge frequency (SEF) 95% decreased during removal of the second testicle compared with baseline recordings. No other significant EEG changes during castration were measured.

CONCLUSIONS

Lidocaine obtunded the EEG changes identified during castration in a previous control study, providing indirect evidence that lidocaine administered peri-operatively was antinociceptive and contributed to anaesthesia during castration.

CONCLUSIONS

The antinociceptive effect of lidocaine combined with its minimal cardiovascular effects indicate a potential use for systemic lidocaine in clinical anaesthetic techniques.

The nucleotide sequence of a 714-bp DNA fragment containing the enterotoxic Escherichia coli (ETEC) adhesive factor 8786 structural gene, designated nfaA, revealed an open reading frame of 498 bp encoding a polypeptide of 166 amino acids. Primer-extension experiments showed that the nfaA gene is within a single transcription unit. No homology was found with the ETEC adhesive factors already sequenced. In contrast, a homology with Salmonella enteritidis fimbrin SEF 14 was observed.

OBJECTIVE

To clarify the somatotopic representation of the tongue secondary somatosensory cortex (SII) in humans.

METHODS

Somatosensory evoked magnetic fields (SEFs) were recorded from nine subjects after stimulating four body sites, left antero (LA) and postero (LP) lateral margins of the tongue, left median nerve at the wrist (Hand), and left tibial nerve at the ankle (Foot).

RESULTS

Clear neural activities were recorded from the bilateral SII in both hemispheres after the four sites were stimulated. The tongue SII for LA and LP was located close to the hand SII and significantly more anterior than the Foot SII. There was no significant difference in the location of dipoles between the LA and LP areas of the tongue SII. The mean peak latencies of the tongue SII for LA and LP were significantly shorter in the hemisphere contralateral to the stimulation than the ipsilateral hemisphere.

CONCLUSIONS

The tongue areas are considered to occupy a small region in SII with insufficient spatial separation to differentiate anterior from posterior areas even using magnetoencephalography which has a higher spatial resolution than electroencephalography (EEG).

CONCLUSIONS

This is the first systematical study to clarify the activated regions in SII following stimulation of the tongue.

At 22 hr after an uncomplicated delivery of a healthy full-term infant, a 26-year-old woman developed toxic-shock syndrome (TSS). A vaginal culture yielded a coagulase-positive Staphylococcus that produced staphylococcal enterotoxin F (SEF) but no other enterotoxins. Breast milk specimens obtained on postpartum days 5, 8, and 11 contained 3.0, 2.5, and 2.0 ng of SEF/ml, respectively. Sera obtained from the mother on postpartum days 4 and 38 had titers (by radioimmunoassay) of antibody to SEF of 1:5 and less than 1:5, a result demonstrating a persisting lack of antibody to SEF after the first episode of TSS; the infant's serum titer of antibody to SEF on day 38 was also less than 1:5. Further longitudinal monitoring of SEF and antibody to SEF in breast milk from this patient is presented. This case is the first isolation of SEF from a body fluid obtained from a patient with TSS further strengthens the association between SEF and TSS.

Bupleuri Radix (Chai-hu in Chinese and Saiko in Japanese) is one of the most important traditional Chinese crude drugs for treating hepatitis malaria and intermittent fever. B. kaoi is one of the Bupleurum spp. families locally found in Taiwan. The effects of saponin-enriched fraction (SEF) from Bupleurum Kaoi in human non-small cell lung cancer A549 cells were investigated in this study. An enhancement in Fas and its two forms of ligands, membrane-bound Fas ligand (mFasL) and soluble Fas ligand (sFasL), was responsible for the apoptotic effect induced by SEF. Taken together, our study suggests that the activity of the Fas/Fas ligand apoptotic system may participate in the antiproliferative activity of SEF in A549 cells.