Neoplastic spindle cells in nasopharyngeal carcinoma show features of epithelial-mesenchymal transition

Luo W‐R, Chen X‐Y, Li S‐Y, Wu A‐B & Yao K‐T
(2012) Histopathology  61, 113–122 Neoplastic spindle cells in nasopharyngeal carcinoma show features of epithelial–mesenchymal transition Aim: To investigate whether the neoplastic spindle cells in nasopharyngeal carcinoma (NPC) are associated with the process of epithelial–mesenchymal transition (EMT). Methods and results: We used immunohistochemistry to analyse the expression of cytokeratin, E‐cadherin, β‐catenin, vimentin, fibronectin, Snail1, Slug and aldehyde dehydrogenase 1 (ALDH1) in 115 cases of NPC in which there were neoplastic spindle cells; in 47 cases a neoplastic squamous cell component was also present. There was no significant difference in the expression of cytokeratin observed in the neoplastic spindle cells (P = 0.644), compared to the squamous component whereas E‐cadherin expression was reduced. By contrast, the expression of β‐catenin, vimentin, fibronectin, Snail1, Slug and ALDH1 was up‐regulated in the spindle cells (all P = 0.000). Furthermore, E‐cadherin expression was associated negatively with β‐catenin (P < 0.001), vimentin (P < 0.001), fibronectin (P < 0.001), Slug (P < 0.001) and ALDH1 (P < 0.001) in neoplastic spindle cells, but did not correlate with Snail1 expression (P =0.093). Conclusions: Our findings demonstrate for the first time that EMT might play an important role in the development of neoplastic spindle cells in NPC.     

Sleep spindle frequency changes during the menstrual cycle

SUMMARY  Five healthy adult women aged 20 to 28 had 12–15 polysomnographic recordings, as well as daily basal body temperature and multiple LH, FSH, estrogen and progesterone measurements taken during a single menstrual cycle. Sleep stages were scored both visually and with a spindle and delta-wave, real-time, automatic analysing system. A cubic growth-curve model showed that the frequency of sleep spindles changed markedly over the menstrual cycle: spindle frequency was lowest about 18 days before onset of menses and highest 3 days before onset of menses. Slow waves did not change. The percentages of Stage 1 and REM sleep showed small changes during the menstrual cycle, and other parameters of visually scored sleep showed no tendency to change. Spindle frequency may reflect the effects of sex hormones on the reticular thalamic nucleus and may be a quantitative marker of premenstrual sleep disturbances.     

Motile human normozoospermic and oligozoospermic semen samples show a difference in double-strand DNA break incidence

BACKGROUND: Among ICSI children de novo structural chromosome aberrations of male descent are increased. Misrepair of double-strand DNA breaks (DSBs) is a prerequisite for such aberrations to occur. To date, no absolute assessment of the number of DSBs in human sperm nuclei after gamete fusion has been described. METHODS: Using man-mouse heterologous ICSI and gammaH2AX immunofluorescent staining, capable of detecting a single DSB, the number of lesions in ICSI selected sperm from normozoospermic men (n = 2) and oligozoospermic patients (n = 3) was quantified. A comparison with a subfertile male mouse model (n = 5) has been made. In addition, the fate of morphologically normal ejaculated immotile sperm after ICSI was examined. RESULTS: A significant increase in the fraction of sperm cells bearing DSBs was found in oligozoospermic semen compared with that from normozoospermic men (P < 0.01). The majority of morphologically normal immotile human sperm showed excess gammaH2AX staining and nuclear disintegration. However, some had a non-deviant DSB pattern. CONCLUSIONS: The increased fraction of DSB-positive sperm in both human and mouse oligozoospermic semen is adding to the surmise that semen from oligozoospermic patients has a reduced chromatin quality, causally related to reduced preimplantation embryo development. The use of ejaculated immotile sperm for in vitro reproduction is debatable due to sperm DNA degradation.     

Mobile phone emission increases inter-hemispheric functional coupling of electroencephalographic alpha rhythms in epileptic patients

It has been reported that GSM electromagnetic fields (GSM-EMFs) of mobile phones modulate - after a prolonged exposure - inter-hemispheric synchronization of temporal and frontal resting electroencephalographic (EEG) rhythms in normal young and elderly subjects (Vecchio et al., 2007, 2010). Here we tested the hypothesis that this can be even more evident in epileptic patients, who typically suffer from abnormal mechanisms governing synchronization of rhythmic firing of cortical neurons. Eyes-closed resting EEG data were recorded in ten patients affected by focal epilepsy in real and sham exposure conditions. These data were compared with those obtained from 15 age-matched normal subjects of the previous reference studies. The GSM device was turned on (45 min) in the "GSM" condition and was turned off (45 min) in the other condition ("sham"). The mobile phone was always positioned on the left side in both patients and control subjects. Spectral coherence evaluated the inter-hemispheric synchronization of EEG rhythms at the following frequency bands: delta (about 2-4 Hz), theta (about 4-6 Hz), alpha1 (about 6-8 Hz), alpha2 (about 8-10 Hz), and alpha3 (about 10-12 Hz). The effects on the patients were investigated comparing the inter-hemispheric EEG coherence in the epileptic patients with the control group of subjects evaluated in the previous reference studies. Compared with the control subjects, epileptic patients showed a statistically significant higher inter-hemispheric coherence of temporal and frontal alpha rhythms (about 8-12 Hz) in the GSM than "Sham" condition. These results suggest that GSM-EMFs of mobile phone may affect inter-hemispheric synchronization of the dominant (alpha) EEG rhythms in epileptic patients. If confirmed by future studies on a larger group of epilepsy patients, the modulation of the inter-hemispheric alpha coherence due to the GSM-EMFs could have clinical implications and be related to changes in cognitive-motor function.     

Automatic analysis of electro-encephalogram sleep spindle frequency throughout the night

A fully automatic method to analyse electro-encephalogram (EEG) sleep spindle frequency evolution during the night was developed and tested. Twenty allnight recordings were studied from ten healthy control subjects and ten sleep apnoea patients. A total of 22 868 spindles were detected. The overall mean spindle frequency was significantly higher in the control subjects than in the apnoea patients (12.5Hz against 11.7Hz, respectively; p<0.004). The proposed method further identified the sleep depth cycles, and the mean spindle frequency was automatically determined inside each sleep depth cycle. In control subjects, the mean spindle frequency increased from 12.0Hz in the first sleep depth cycle to 12.6Hz in the fifth cycle. No such increase was observed in the sleep apnoea patients. This difference in the spindle frequency evolution was statistically significant (p<0.004). The advantage of the method is that no EEG amplitude thresholds are needed.     

Sleep evoked delta frequency responses show a linear decline in amplitude across the adult lifespan

Aging is associated with many changes in sleep, with one of the most prominent being a reduction in slow wave sleep. Traditional measures of this phenomenon rely on spontaneous activity and typically confound the incidence and amplitude of delta waves. The measurement of evoked K-complexes during sleep, enable separate assessment of incidence and amplitude taken from the averaged K-complex waveform. The present study describes data from 70 normal healthy men and women aged between 19 and 78 years. K-Complexes were evoked using short auditory tones and recorded from a midline array of scalp sites. Significant reductions with age were seen in the amplitude of the N550 component of the averaged waveform, which represents the amplitude of the K-complex, with linear regression analysis indicating approximately 50% of the variance was due to age. Smaller, yet still significant reductions were seen in the ability to elicit K-complexes. The data highlight the utility of evoked K-complexes as a sensitive marker of brain aging in men and women.     

Granular cell astrocytomas show a high frequency of allelic loss but are not a genetically defined subset

Granular cell astrocytomas (GCA) are an uncommon morphologic variant of infiltrative glioma that contains a prominent population of atypical granular cells. As a rule, they are biologically aggressive compared to similar tumors without granular features. We sought to determine whether GCAs possess distinct genotypic alterations that might reflect their unique morphology or clinical behavior. Eleven GCAs occurring in 7 men and 4 women ranging in age from 46 to 75 years were investigated for genetic alterations of known significance in glial tumorigenesis, including LOH at 1p, 9p, 10q, 17p, and 19q, point mutations of TP53, deletions of p16(CDKN2A) and p14ARF, as well as EGFR amplifications. Tumors included had an infiltrative growth pattern and consisted of large, round cells packed with eosinophilic, PAS-positive granules that varied in quantity, ranging from 30 to 100% of tumor cells. Three tumors were of WHO grade II, one was grade III, and 7 were grade IV lesions. Overall, the tumors showed higher frequencies of LOH at 1p, 9p, 10q, 17p, and 19q than typical infiltrating astrocytomas of similar grades. Losses on 9p and 10q occurred in nearly all cases, including low grade lesions. TP53 mutations were identified in 2 grade IV GCAs, while combined p14ARF and p16(CDKN2A) homozygous deletions were noted in only one grade IV lesion. None showed EGFR amplification. We found no genetic alterations specific for GCA. Instead, it appears that granular cell change occurs across genetic subsets. The high frequency of allelic loss, especially on 9p and 10q, may confer aggressive growth potential and be related to their rapid clinical progression.     

Normal colon tissue and colon carcinoma show no difference in heparanase promoter methylation

Introduction

Heparanase, the sole heparan sulfate degrading enzyme, has a role in cellular invasion. Accordingly, a large number of studies have demonstrated an association between heparanase expression and tumor stage and patients' prognosis. In colon carcinoma, heparanase shows increased expression in tumor compared to normal tissue and its expression correlates with the presence of metastasis. One of the regulatory mechanisms of heparanase expression is de-methylation on its promoter. In the present study we evaluated the role of heparanase promoter methylation in colon carcinoma.

Material and methods

Analysis of heparanase promoter methylation was done on 32 samples of colon carcinoma as well as 30 samples of normal colonic mucosa. DNA was extracted from FFPE tissue and subjected to bisulfite conversion. The relative fraction of methylated and unmethylated DNA was evaluated using quantitative real-time PCR.

Results

The fraction of methylated DNA was 1 ± 3.4% in the colon carcinoma group, and 2.5 ± 3.3% in the normal colon group (P = 0.11). Only one case in the normal group and one case in the tumor group showed more than 10% methylation in the heparanase promoter.

Conclusion

We did not find any significant difference in heparanase promoter methylation between colon carcinoma and normal colonic mucosa, suggesting that heparanase overexpression in colon carcinoma is mediated by other mechanisms.     

Effect of frequency difference on sensitivity of beats perception

Two vibrations with slightly different frequencies induce the beats phenomenon. In tactile perception, when two pins of different frequencies stimulate the fingertips, an individual perceives a beats caused by a summation stimulus of the two vibrations. The present study demonstrates experimentally that humans can perceive another vibration based on the beats phenomenon when two tactile stimuli with slightly different frequencies are stimulated on the finger pad with a small contactor in different locations at the same time. Moreover, we examined the amplitude of the detection threshold to be able to perceive beats phenomenon on the index finger with 5 carrier frequency (63.1, 100, 158.5, 251.2, and 398.1 Hz) and 4 beats frequency (2.5, 3.98, 6.31, and 10 Hz) when two stimuli 1 mm distance apart are vibrated at a slightly different frequency. From the experiments, it is concluded that the amplitude threshold to be able to perceive beats decreases as the standard frequency increases under 398 Hz. Furthermore, from comparing the absolute detection threshold and beats detection threshold, as the carrier frequency increases, the required amplitude at two pins for the detection of beats decreases compared to absolute vibration.     

Basophils from patients with allergic asthma show a primed phenotype.

BACKGROUND: IL-3, IL-5, and GM-CSF are not able to induce histamine release in purified basophils of nonallergic donors. However, we have recently found that preincubation with 2 micromol/L thapsigargin, which induces a rise in intracellular free calcium ions, renders human basophils extremely sensitive for IL-3, IL-5, or GM-CSF, leading to enhanced histamine release. Histamine release was also induced in the reverse order (first cytokine and then thapsigargin). OBJECTIVE: Because these cytokines are supposed to be increased in allergic inflammation, we examined whether basophils of patients with allergic asthma showed an enhanced response to thapsigargin. METHODS: We measured the histamine release induced by thapsigargin in a group of allergic asthmatic subjects (n = 24) and compared this response with those of 3 control groups. The control groups consisted of healthy control subjects (group 1, n = 21); patients with a nonallergic, nonasthmatic lung disease (group 2, n = 22); and patients with nonallergic asthma (group 3, n = 9). RESULTS: There was no difference in spontaneous histamine release. Also, no significant difference in histamine release was found when anti-IgE or formyl-methionyl-leucyl-phenylalanine was used as a stimulus. Histamine release induced by IL-3 alone or a combination of IL-3 and thapsigargin also did not differ. In contrast, basophils from the group with allergic asthma showed a significantly higher percentage of histamine release induced by thapsigargin (38.2% +/- 13.2%) than did basophils from the 3 control groups (healthy control subjects, 22.5% +/- 6.9%; subjects with lung disease, 24.9% +/- 8.9%; subjects with nonallergic asthma 15.0% +/- 3.0%; all mean +/- SD). CONCLUSION: These data indicate that basophils in peripheral blood of subjects with allergic asthma have a primed phenotype and that thapsigargin-induced histamine release is a practical tool to study this phenomenon.     

Apnea revisited: a longitudinal follow-up

Volunteers aged 65 years and over who were previously studied underwent follow-up interviews and sleep recordings several years later (mean follow-up, 4.6 years). Modest correlations between the initial and subsequent recordings were found for apnea index, hypopnea index, and respiratory disturbance index. Sleep apnea indices did not increase over time; however, individual subjects showed great variability in amount of apnea. Periodic leg movement indices, on the other hand, significantly increased. Subjective reports indicated a significant increase in the amount of time spent awake and a significant decrease in the amount of loud snoring.     

No show is a major problem in planning appointments for infertility patients

We examined the pattern of bookings for and actual visits undertaken by patients attending the Infertility Clinics at the Rotunda Hospital Dublin as well as the accuracy of systems in place for recording such events. The data provided by the Hospital Patient Administration System (PAS), clinic worksheets and clinic record chart of first visit couples attending initially over a 12 month period 1995/1996 and their subsequent records to February 1998 was analysed. Comparable figures were provided from the corresponding private clinic over the same time frame. Sixty-nine percent had already had investigations and treatment elsewhere. Discrepancies were noted between the PAS and manual systems as confirmed by the patient records. The manual proved more accurate. We found 32% (88 of 276) patients failed to turn up for a first appointment compared with 17% of private patients. Un-notified no-show provides great logistic difficulties in planning clinic management. Waiting lists grow, staff and patients alike are greatly inconvenienced. This apparent recklessness makes the Charter of Rights for Hospital Patients difficult to comply with. A possible answer is to demand reconfirmation near to the clinic date and in it's absence reassign the appointment to another. Population education is clearly needed.     

Magnitude of oscillations in the response of Ia muscle spindle endings under a static gamma stimulation of increasing frequency

Under static gamma stimulations, Ia afferents may discharge in a highly irregular way or may be driven. However, the genesis of the highly irregular form of discharge is unclear. We offer an interpretation of irregular discharge behavior. Twenty-three primary (Ia) muscle spindle afferents from the tibial anterior muscle of the cat were subjected to static gamma stimulation, the stimulation frequency increasing linearly from 2 to 110/s. In addition, 17 of the spindle afferents were subjected to two different prestretch values of the muscle while the static gamma fiber was now subjected to constant frequency stimulation at five different stimulation frequencies ranging from 9.4 to 95/s. The responses of the Ia afferents to the static gamma stimulation were presented through discharge patterns that were constructed by the frequencygram method and were subjected to computer analysis, by means of which the Ia responses were evaluated quantitatively. Two groups of static gamma stimulations were identified. The first group of gamma stimulations leads in the Ia response to highly irregular discharging within a broad discharge band. This highly irregular discharging resolves into regular oscillatory responses of large magnitude occurring in the rhythm of the gamma stimuli. According to this observation, the highly irregular discharges result from the fact that the Ia afferent generates more than one action potential per gamma stimulus. The second group of gamma stimulation leads in the Ia response either to driving of the action potentials in the rhythm of the gamma stimulation frequency or of submultiples of it or to irregular discharging within a smaller discharge band. Under the two latter conditions, oscillatory Ia responses of small magnitude occurring in the rhythm of the gamma stimuli are proved to be generated by the Ia afferents. The results are explained in terms of the strength of contraction of the polar parts and the resulting stretch of the sensory parts of the intrafusal muscle fibers that are responsible.     

Analysis of response properties of deefferented mammalian spindle receptors based on frequency response.

Sinusoidal responses of primary and secondary endings in deefferented spindles of anesthetized cats were studied over the low-frequency range 0.001-0.1 Hz. Stretch amplitudes were chosen conservatively small (25-100 mum peak-to-peak) so as to lie within the linear region. 1. At 0.1 Hz average sensitivity was 350 pps/mm for primary endings and 80 pps/mm for secondary endings. Sensitivity fell to lower values at lower frequencies, but even at 0.001 Hz, corresponding to 17 min/cycle, sensitivity remained elevated above static values determined with large stretches. Phase lead varied from 5 to 50 degrees and, in the case of primary endings, tended to be greater at lower frequencies. 2. Except for the different scaling factors, the only apparent difference between the frequency responses of primary and secondary endings was a tendency for primary endings to show a greater phase lead over the range 0.001-0.01 Hz. 3. Dynamic responsiveness was assessed theoretically from frequency-response data by calculating responses to ramps at various velocities. Over most of the velocity range dynamic responses were not proportional to velocity. The greater dynamic responsiveness of primary endings during large (6 mm) ramp stretches might be related to frequency response below 0.01 Hz. 4. Certain aspects of dynamic responsiveness to large ramps (6 mm) were accounted for by assuming all phases of responses were attenuated by 25 dB in the case of primary endings and 20 dB in the case of secondary endings. The nonlinearity responsible for attenuation appears to occur at an early stage in the sensory process. 5. Comparison of individual responses to slow ramps with predictions based on linear theory indicated the presence of abrupt departures from linearity for both primary and secondary endings.     

Renal transplant patients show variations in their self-reactive repertoires: a serial study

We addressed the question of whether allo-transplantation (Tx) induces breakdown of tolerance to self-antigens or alteration of the autoreactive T cell repertoire in humans. The serial variation of T cell autoreactivity was studied in the peripheral blood of 12 renal transplant patients, by autologous limiting dilution assay and autologous mixed lymphocyte reaction. Ten of 12 patients presented a positive response in autologous peripheral blood mononuclear cells in the post-Tx period, in contrast to four of 12 patients before Tx (P = 0.038). Multi-hit kinetics was found in 57% of the assays analyzed, indicating frequent regulatory control of the autologous response. Quantitative analysis performed in eight patients showed an increase in precursor frequency at >1 year post-Tx in five patients. These data indicate that autoreactivity increases or develops following Tx, in humans. Post-Tx events such as alloreactivity, infections or immunosuppression could interfere with the balance of autoreactive and regulatory cells, leading to changes in the T cell repertoires to self-antigens and eventually breakdown of self-tolerance. Further investigation is needed to elucidate whether post-Tx autoreactivity contributes to rejection, plays a regulatory role over alloreactivity or both, at separate times.     

Sleep spindle activity changes in patients with affective disorders

Various polysomnographic sleep patterns are associated with affective disorders, but very little is known about sleep spindle characteristics in adult depression. In primary endogenous depressive male patients (unipolar, UP, and bipolar, BP) with comparable depression scores and in normal control subjects recorded during 3 consecutive nights, no night effect was observed on the sleep variables investigated except for REM latencies of stages 1 and 2. Stage 2 duration and variables related to sleep spindle characteristics (the number and the density of spindles of 1/2 s; the number and the density of full spindles of stage 2 over the 3 nights) were significantly lower in depressed patients than in control subjects, the mean number of spindles being lower in UP than in BP patients. Sleep spindle measures were clearly negatively correlated with age in the overall group (i.e., depressed plus control subjects). They were also negatively correlated with the REM latencies of stages 1 and 2 in BP depressed patients, whereas this relation was not observed in UP patients.     

Visceral and post-Kala-Azar dermal leishmaniasis isolates show significant difference in their in vitro drug susceptibility pattern

Visceral leishmaniasis (VL) remains a major health problem in old world, and India accounts for half of the world burden. The widespread emergence of resistance to standard drug in India poses a major obstacle in the control of leishmaniasis. Post-Kala-Azar dermal leishmaniasis (PKDL) is considered as main source of drug resistance. Experimental data indicate that resistance against newer drugs is also imminent. Therefore, in vitro studies were carried out to test minimum parasiticidal concentration of five conventional and newly introduced anti-leishmanial drugs against 20 field isolates of Leishmania donovani obtained from visceral and post-Kala-Azar dermal leishmaniasis patients of India. Study revealed wide range of variation in minimum inhibitory concentration of sodium antimony gluconate (SAG). PKDL isolates displayed significantly lower susceptibility to SAG and miltefosine than VL isolates with P value of 0.0006 and 0.0243, respectively. All clinical isolates had higher IC₅₀ value for paromomycin and miltefosine as compared to reference strain indicating their vulnerability to develop unresponsiveness. However, isolates were uniformly susceptible to pentamidine and amphotericin B. The results of gene expression analysis of AQP1 were largely in agreement with phenotypic drug sensitivity results. Interestingly, significant down-regulation of AQP1 was observed in PKDL isolates as compared to VL isolates indicating their increased propensity for drug unresponsiveness. However, no significant difference in mRNA expression of LdMT and LdRos3 gene was found for two groups. The present study unravels valuable baseline scientific data showing variation in the drug susceptibility pattern in the L. donovani isolates. The information might have impact on the management and control of Indian visceral leishmaniasis.