CD133 induces tumour-initiating properties in HEK293 cells

The pentaspan protein CD133 (Prominin-1) is part of the signature of tumour-initiating cells for various cancer entities. The aim of the present study was to investigate the impact of ectopic CD133 expression on tumourigenic properties of otherwise CD133-negative, non-tumourigenic cells in vitro and in vivo. CD133 was stably transfected into human embryonic kidney 293 (HEK293) which was then sorted for the expression of CD133. The effects of CD133 on cell proliferation were assessed upon standard cell counting of sorted cells at various time points. Severe combined immunodeficient (SCID) mice (n?=?30) were injected with HEK293 CD133^high and CD133^low transfectants (5?×?10³, 1?×?10⁵, or 5?×?10⁶ cells per injection). The expression of CD133, Ki67, CD44s, CD44v6, and EpCAM was analysed upon immunohistochemical staining of cryosections with specific antibodies. In vitro, ectopic expression of CD133 did influence neither cell proliferation nor cell cycle distribution of otherwise CD133-negative HEK293 cells. However, CD133^high cells generated tumours in vivo in SCID mice with at least 1,000-fold increased frequency compared to CD133^low cells. Tumour load was also significantly increased in CD133^high cells as compared to those tumours formed by high numbers of CD133^low cells. Immunohistochemistry stainings disclosed no changes in Ki67, CD44s, CD44v6, or EpCAM once tumours were formed by either cell type. CD133 induces tumour-initiating properties in HEK293 cells in vivo and is potentially involved in the regulation of tumourigenicity. Future research will aim at the elucidation of molecular mechanisms of CD133-induced tumourigenicity.     

Evaluation of the prophylactic use of mitomycin-C to inhibit haze formation after photorefractive keratectomy

PURPOSE: To evaluate the results of the prophylactic use of mitomycin-C to inhibit haze formation after excimer laser photorefractive keratectomy (PRK) for medium and high myopia in eyes that were not good candidates for laser in situ keratomileusis (LASIK). SETTING: Carones Ophthalmology Center, Milan, Italy. METHODS: This prospective randomized masked study comprised 60 consecutive eyes (60 patients). The inclusion criteria were a spherical equivalent correction between -6.00 and -10.00 diopters (D) and inadequate corneal thickness to allow a LASIK procedure with a residual stromal thickness of more than 250 microm. The eyes were divided into 2 groups according to the randomization protocol. After PRK, the study group eyes were treated with a single intraoperative dose of mitomycin-C (0.2 mg/mL), applied topically with a soaked microsponge placed over the ablated area and maintained for 2 minutes. The control eyes did not receive this treatment. Refraction, uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), and slitlamp evidence of corneal opacity (haze) or other visible complications were evaluated. RESULTS: No toxic or side effects were encountered postoperatively. No study group eye had a haze rate higher than 1 during the 6-month follow-up; 19 eyes (63%) in the control group did (P =.01). At 6 months, the between-group difference in the refractive outcome was statistically significant (P =.05), with 26 study group eyes (87%) and 14 control eyes (47%) within +/-0.50 D of the attempted correction. No study group eye had a BCVA loss during the follow-up; 7 control eyes had lost 1 to 3 lines at 6 months (P =.0006). CONCLUSIONS: The prophylactic use of a diluted mitomycin-C 0.02% solution applied intraoperatively in a single dose after PRK produced lower haze rates, better UCVA and BCVA results, and more accurate refractive outcomes than those achieved in the control group.     

Inhibitory activity of xanthine oxidase by fractions Crateva adansonii

ObjectiveTo study the inhibitory effect of various extracts from Crateva adansonii (C. adansonii) used traditionally against several inflammatory diseases such as rheumatism, arthritis, and gout, was investigated on purified bovine milk xanthine oxidase (XO) activity.MethodsXanthine oxidase inhibitory activity was assayed spectrophotometrically and the degree of enzyme inhibition was determined by measuring the increase in absorbance at 295 nm associated with uric acid formation. Enzyme kinetics was carried out using Lineweaver-Burk plots using xanthine as the substrate.ResultsAmong the fractions tested, the chloroform fraction exhibited highest potency (IC₅₀ 20.2±1.6 μg/mL) followed by the petroleum ether (IC₅₀ 30.1±2.2 μg/mL), ethyl acetate (IC₅₀ 43.9±1.4 μg/mL) and residual (IC₅₀ 98.0±3.3 μg/mL) fractions. The IC₅₀ value of allopurinol used, as the standard was 5.7±0.3 μg/mL.ConclusionsEnzyme inhibition mechanism indicated that the mode of inhibition was of a mixed type. Our findings suggest that the therapeutic use of these plants may be due to the observed Xanthine oxidase inhibition, thereby supporting their use in traditional folk medicine against inflammatory-related diseases, in particular, gout.     

Electrical Performance and Automatic Capture Characteristics of a 3.5-mm2 Passive Fixation Lead during 1-Year Follow-Up

Background: Bipolar low polarization electrodes are recommended for a regular AutoCapture™ (St. Jude Medical, Inc., Sylmar, CA, USA) function in order to effectively detect the evoked response (ER) signal. The objective of this national multicenter registry was to evaluate the electrical performance and the AutoCapture™ characteristics of the bipolar ventricular pacing lead IsoFlex S, model 1636T or 1646T (St. Jude Medical), in combination with single- and dual-chamber pacemakers.
Methods: Ventricular pacing and sensing thresholds, lead impedance, ER amplitude, and polarization signals were measured at discharge and routine follow-up visits after 1, 3, 6, 9, and 12 months. AutoCapture™ activation was recommended based on the results of the ER sensitivity test.
Results: Of the 252 patients initially included, 109 (43%) have completed the follow-up. The mean ventricular pacing threshold was 0.43 ± 0.19 V at discharge and 0.68 ± 0.32 V at 12 months postimplant. The values for the ventricular sensing threshold were between 9.51 ± 4.12 and 9.99 ± 4.09 mV at discharge and at the 12-month follow-up. The unipolar lead impedance decreased from 533 ± 94 to 476 ± 73 ohms during the follow-up. The mean ER amplitude was 16.47 ± 6.70 mV at discharge and 17.42 ± 7.43 mV after 12 months, and the corresponding mean polarization signals were 0.59 ± 1.00 and 0.74 ± 1.24 mV, respectively. AutoCapture™ activation was recommended in at least 95% of the patients investigated over the 12-month follow-up.
Conclusion: The bipolar ventricular pacing lead IsoFlex S 1636/1646T shows a good electrical performance and is mostly compatible with the AutoCapture™ algorithm.     

Evaluation of tracheobronchial lesions with spiral CT: comparison between virtual endoscopy and bronchoscopy]

INTRODUCTION: The aim of this study is to describe the scanning parameters for virtual bronchoscopy in the evaluation of the tracheobronchial tree and to compare the results of this examination with the endoscopic findings. MATERIAL AND METHODS: 27 patients with tracheobronchial neoplasms suspected at preliminary clinical and chest film findings or postoperative follow-up for malignant disease were evaluated with spiral CT of the chest and bronchoscopy. Virtual endoscopy was performed on the pulmonary volume involved by the lesion, using narrow axial images (thickness 2 mm, table index 3 mm, reconstruction index 1 mm.) so as to obtain MPR, MIP and 3D reconstructions with 3D Endo Vew program (Philips Medical System, Eindhoven, Holland). We compared these reconstructions and the findings the normal spiral CT scanning with the corresponding endoscopic examinations. RESULTS: In all patients we were able to study the lobar and segmental bronchi in all patients and in 2 we also evaluated the subsegmental bronchi. 25 lesions in 23 patients were shown by virtual endoscopy (8 occlusions, 8 stenosis, 5 compressions, 3 flogosis with endobronchial mucus, 1 bronchocele) and in 4 patients the examinations were negative. The bronchoscopy was negative in 4 patients and positive in 23 patients with 25 lesions, but we had agreement in 23/27 patients (85,1%). In 2 patients virtual endoscopy showed the lesions in a different bronchus compared to bronchoscopy. In one patient we interpreted the obstruction as neoplastic instead of mucus inside the bronchi and in the last patient bronchoscopy was not performed due to his old age and the virtual endoscopy showed total obstruction of a segmental bronchus. DISCUSSION AND CONCLUSIONS: The results show that virtual endoscopy can study the tracheobronchial tree as far as the segmental bronchi, and sometimes also the subsegmental bronchi and the bronchi below a closed obstruction. In addition, it can evaluate the extraluminal location of the lesions. For these reasons virtual endoscopy provides a road map for bronchoscopy as a guide for transbronchial biopsy and for endobronchial treatment planning. The limitation of this technique is its inability to evaluate the mucosal surface and distinguish flogosis from neoplastic lesions by biopsy. It can be used however in the postoperative follow-up both for cancer and transplant, when immediate biopsy is not necessary.     

细粒棘球蚴重组抗原B的表达提取及其血清学检测初探

用基因工程技术制备出的细粒棘球绦虫重组抗原B（rAgB）表达载体,经诱导表达、亲和层析纯化获得具生物活性的重组蛋白质rAgB,用Western-Blot法检测病人血清。结果显示rAgB敏感性为91．6％（44／48）,特异性为93．8％（30／32）,其中10例泡球蚴（AE）病人及10例肿瘤病人血清均无交叉反应。说明rAgB具有较高的敏感性及特异性,可用于包虫病的常规血清学诊断,rAgB在宿主菌JM109内稳定表达,因此可在实验室内大量制备用于血清学诊断的rAgB。     

Ocular penetration and pharmacokinetics of cefotaxime: an experimental study

Cefotaxime levels in the cornea and the aqueous humour were assayed after local topical application and subconjunctival injection in healthy and infected corneas. The results in infected corneas were analysed pharmacokinetically. The absorption and elimination of cefotaxime within 2 hours after topical administration are adequately predicted by a two-compartment model in which one compartment represents the cornea and the other the aqueous humour, with the elimination taking place from the latter. The transfer coefficient kc (cornea-aqueous) was estimated as 1.452 h-1 and the return transfer coefficient kca (aqueous-cornea) as 0.287 h-1. After subconjunctival injection the levels measured exhibit bi-exponential kinetics, with the apparent absorption constant Ka = 3.816 h-1.     

Circadian periodicity of heart rate variability in hospitalized angor patients

Abstract The relationship between unstable angor (angina) and circadian periodicity of heart rate variability (HRV) was explored in a group of patients hospitalized in a coronary care unit (CCU). Patients were classified as normal (whose symptoms had non-cardiovascular origin, n=8), moderate angor (n=13) and severe angor (n=11). A fourth group of ambulatory healthy volunteers (n=12) was included. Individual 24 h Holter records were analyzed, mean RR and standard deviation of RR (SDNN) being obtained from 1 h-length windows. For frequency domain analysis, 5 min-length windows were employed. The spectral components analyzed were total power (spectral power between 0.01 and 0.5 Hz), low frequency power (LF: power between 0.04 and 0.15 Hz), and high frequency power (HF: power between 0.15 and 0.4 Hz). In addition, LF to HF areas ratio (L/H) was computed. Mesor, amplitude and acrophase for every 24 h rhythm were calculated by cosinor analysis.As compared to ambulatory controls, admission to the CCU diminished amplitude and phase-delayed the circadian oscillation of most HRV parameters, except for SDNN. Moderate angor patients showed decreased amplitude of RR and L/H and augmented amplitude of SDNN when compared to normal hospitalized subjects. A phase delay of about 1.5 h for RR intervals and a phase advance of 3.5–6 h for LFA and SDNN were found in the moderate angor group when compared to normal. Amplitude of 24 h variation of total power decreased in severely angor patients and the circadian oscillation of HF (an indicator of vagal control on the heart) became free running. A phase delay of 2.5 h in SDNN acrophase was found in severely affected patients when compared to moderate. The results indicate that severity of unstable angor correlates with desynchronization of parasympathetic control of heart rate.     

Safety and efficacy of a new recombinant FVIII formulated with sucrose (rFVIII–FS) in patients with haemophilia A: a long-term, multicentre clinical study in Japan

The recombinant full-length FVIII product Kogenate has been reformulated using sucrose (rFVIII-FS) instead of human serum albumin as a stabiliser in purification and formulation. The in vivo recovery, haemostatic efficacy, and safety of rFVIII-FS were investigated in 20 previously treated patients with severe or moderate haemophilia A for > or = 24 weeks. In vivo recoveries of 73.5 +/- 16.3%, 78.4 +/- 16.1%, and 82.8 +/- 23.9% after the initial infusion of 50 IU kg(-1) rFVIII-FS and at weeks 12 and 24, respectively, showed no significant changes over time. A total of 1115 infusions (mean dose 24.1 +/- 8.4 IU kg(-1)) were included in the analysis of haemostatic efficacy. One (80.5%) or two (8.2%) infusions achieved adequate haemostasis in 88.7% of all bleeding episodes, and haemostatic efficacy was judged 'excellent' or 'good' in 749 of 764 episodes (98.0%). The haemostatic efficacy was judged as 'excellent' or 'good' in 924 of 1115 (82.9%) infusions. Twenty-one adverse events were observed in 12 patients in the total 1541 infusions included in the safety analysis. Causality with respect to rFVIII-FS could not be ruled out in three events in one HIV-negative patient: elevated CD4(%), decreased CD8(%), and elevated CD4/CD8 ratio. No FVIII inhibitor development was observed in any patient. ELISA assay testing for antibodies to rFVIII, baby hamster kidney cell (BHK) protein, and murine IgG were all negative. These results show that rFVIII-FS is a safe and effective for long-term treatment of patients with haemophilia A.