Amyloid fibril formation by human stefin B: influence of pH and TFE on fibril growth and morphology.

As shown before, human stefin B (cystatin B) populates two partly unfolded species, a native-like state at pH 4.8 and a structured molten globule state at pH 3.3 (high ionic strength), from each of which amyloid fibrils grow. Here, we show that the fibrils obtained at pH 3.3 differ from those at pH 4.8 and that those obtained at pH 3.3 (protofibrils) do not transform readily to mature fibrils. In addition we show that amorphous aggregates are also a source of fibrils. The kinetics of amyloid fibril formation at different trifluoroethanol (TFE) concentrations were measured. TFE accelerates fibril growth at predenaturational concentrations of the alcohol. At concentrations higher than 10%, the fibrillar yield decreases proportionately as the population of an all alpha-helical, denatured form of the protein increases. At an optimum TFE concentration, the lag and the growth phases are observed, similarly to some other amyloidogenic proteins. Morphology of the protein species at the beginning and the end of the reactions was observed using atomic force microscopy and transmission electron microscopy. Final fibril morphologies differ depending on solvent conditions.     

Concomitant radiotherapy with mitomycin C and bleomycin compared with radiotherapy alone in inoperable head and neck cancer: final report

Purpose: To compare the efficacy of concomitant irradiation with mitomycin C and bleomycin in patients with inoperable head and neck carcinoma with radiotherapy alone.

Methods and materials: Between March 1991 and December 1993, 64 patients with inoperable head and neck carcinoma (41 with oropharyngeal site) were randomized to radiotherapy alone (group A) or radiotherapy combined with simultaneous application of mitomycin C and bleomycin (group B). In both groups patients were irradiated five times weekly with 2 Gy to a total dose of 66–70 Gy. The planned concomitant treatment in group B was: bleomycin 5 units twice a week IM, total dose 70 units, mitomycin C 15 mg/m² IV after delivery of 10 Gy, and 10 mg/m² IV on the last day of radiotherapy. To enhance the effect of these two drugs, patients received also nicotinamide, chlorpromazine, and dicoumarol.

Because significantly better results were achieved in arm B for patients with inoperable oropharyngeal carcinoma, the study was closed and such patients were after December 1993 routinely treated with the combined therapy (as in arm B). Until October 1996, we treated and followed up 48 such consecutive patients.

Results: Median follow-up of our study patients is 42 months. Complete remission (CR) rate in group A was 31% and in group B 59% (p = 0.04); disease-free survival (DFS) in group A was 8% and in group B 37% (P = 0.01); and overall survival (OS) was 7% in group A and 26% in group B (p = 0.08). CR rate for patients with oropharyngeal carcinoma was 29% in group A (N = 21) and 75% in group B (N = 20) (p = 0.007); DFS in group A was 10% and in group B 48% (p = 0.001); and the OS was 10% in group A and 38% in group B (p = 0.019). In patients with inoperable oropharyngeal carcinoma treated after December 1993, complete remission was achieved in 32/48 (67%, 95% CI: 52%–80%). DFS at the median follow-up of 14 months was 60% (95% CI 43–77%) and OS 58% (95% CI 42–74%).

Conclusion: From the results of our study it seems that the concomitant treatment significantly improves CR rate, DFS, and OS in patients with inoperable oropharyngeal carcinoma in comparison with radiotherapy alone.     

Change of phonation control after cochlear implantation.

OBJECTIVE: To assess the influence of acquired auditory control on some voice parameters in deaf children and adults after cochlear implantation. STUDY DESIGN: Prospective clinical study. SETTING: Tertiary referral center. PATIENTS: Twenty-nine prelingually deafened children and 11 postlingually deafened adults. INTERVENTIONS: The samples of a vowel /a/ were analyzed with an Multi-Dimensional Voice Program (Kay Elemetrics Corporation, Lincoln Park, NJ) before and 6 to 12 months after the cochlear implantation. MAIN OUTCOME MEASURES: The average fundamental frequency (F0), the short-term variation of F0 (JIT) and the amplitude (SH), the very long-term variation of F0 (vF0) and the amplitude (vAm), and the noise-to-harmonic ratio (NHR) were determined and compared for both age groups. The results of the acoustic analysis performed before the implantation were compared with the results after the implantation for children and adults. RESULTS: Significantly greater JIT, SH, vF0, and vAm were detected in the children than in the adults before and after the implantation. The prelingually deafened children significantly improved the control of their phonation after 6 to 12 months' use of the cochlear implant (JIT: p=0.014, SH: p=0.011, vF0: p=0.014, vAm: p=0.031). In the postlingually deafened adults, no significant improvement was found in any of the studied voice parameters after the implantation. F0 showed little or no change after the implantation in children and adults. CONCLUSION: As expected, the voice quality of the prelingually deafened children was significantly worse than that of the postlingually deafened adults. After cochlear implantation, the children significantly improved their short-term and long-term F0 and amplitude variability. In adults, no significant improvement was detected. We suppose that the improvement is a consequence not only of the acquired hearing control but also of the adaptation ability of neuromuscular phonation control and the maturing of these control mechanisms in children. In adults, better phonation quality in general and lesser improvement after the implantation can be the results of well-developed and stable phonation patterns.     

Sensing surface morphology of biofibers by decorating spider silk and cellulosic filaments with nematic microdroplets

Probing the surface morphology of microthin fibers such as naturally occurring biofibers is essential for understanding their structural properties, biological function, and mechanical performance. The state-of-the-art methods for studying the surfaces of biofibers are atomic force microscopy imaging and scanning electron microscopy, which well characterize surface geometry of the fibers but provide little information on the local interaction potential of the fibers with the surrounding material. In contrast, complex nematic fluids respond very well to external fields and change their optical properties upon such stimuli. Here we demonstrate that liquid crystal droplets deposited on microthin biofibers—including spider silk and cellulosic fibers—reveal characteristics of the fibers’ surface, performing as simple but sensitive surface sensors. By combining experiments and numerical modeling, different types of fibers are identified through the fiber-to-nematic droplet interactions, including perpendicular and axial or helicoidal planar molecular alignment. Spider silks align nematic molecules parallel to fibers or perpendicular to them, whereas cellulose aligns the molecules unidirectionally or helicoidally along the fibers, indicating notably different surface interactions. The nematic droplets as sensors thus directly reveal chirality of cellulosic fibers. Different fiber entanglements can be identified by depositing droplets exactly at the fiber crossings. More generally, the presented method can be used as a simple but powerful approach for probing the surface properties of small-size bioobjects, opening a route to their precise characterization.Natural microfilaments produced by plants, insects, or spiders are fascinating materials not just because of their specific properties such as wear resistance, elasticity, tensile strength, and toughness (1–5) but also because of their microorganization (6–9). Their macroscopic properties can match properties of materials like kevlar but are at the same time biocompatible and biodegradable (10). These fascinating macroscopic properties actually originate from bulk and surface properties of the fibers (1). The chemical composition of the threads combined with their morphology determines the final properties of the material (11–13). The mechanical properties of the spider fibers are determined by the existence of a lyotropic liquid crystalline phase, from which the threads are drawn (14). Such silks are known to include nanoscale networks of defects and cavities that yield surface structures notably dependent on the spider species (3). These differences do not affect much the mechanical performance of the fibers (1, 3, 5). From a technological perspective, many attempts have been made to reproduce these natural bionetworks (15–17). In fact cellulose-based fibers with few micrometers of diameter, produced by electrospinning, can also acquire different morphologies depending upon the processing conditions, giving diverse features of the final threads and mats (18). Therefore, probing the surface structure of the microfibers is crucial for a complete understanding of their individual and interthreaded properties.From another perspective, nematic complex fluids are materials which are inherently responsive to diverse external stimuli, notably including diverse surface interactions which in the literature are known as the surface anchoring (19). Being effectively elastic materials, the orientational order of nematics responds on long, typically micrometer scales (20–22), which results in a spatially varying birefringence that can be optically detected (23). Recently, it was demonstrated that glass fibers induce numerous defects in a well-aligned nematic liquid crystal cell and thus provide a simple illustration of topological phenomena (24). It is also known that liquid crystal droplets can considerably change their structure by the action of otherwise imperceptibly small external stimuli (21). Pierced nematic and chiral nematic droplets develop defects that can be controlled by the liquid crystal elasticity, chirality, and surface boundary conditions (25, 26) indicating exceptional sensitivity. Therefore, to generalize, putting nematics into contact with diverse surfaces (18, 27) can be used as a simple but very powerful technique to detect the surface properties of microobjects such as biological fibers.In this paper we demonstrate the surface morphology sensing of biorelevant fibers, including spider silk and cellulosic microfibers, by nematic droplets that are sprayed onto the fibers. Specifically, we explore the chiral and achiral nature of the fiber’s surface and the in-plane or perpendicular alignment fields the fibers impose on the nematic. Droplets with degenerate in-plane and perpendicular alignment of the nematic at their free surfaces are explored, combining experiments and numerical modeling, to allow for tuning of the sensing precision. Further, the entanglement sites of the fiber webs are explored, with the droplets deposited at the sites clearly revealing contact, noncontact, and entangled morphologies.     

His bundle pacing in a young child guided by electroanatomical mapping

Pediatric patients with complete congenital atrio‐ventricular (AV) block are generally exposed to life‐long dyssynchronous right ventricular (RV) pacing. His bundle pacing (HBP) is an alternative method of pacing that better restores physiological ventricular activation which could prevent pacing‐induced cardiomyopathy. We present a case of a 5‐year‐old child with complete AV block who underwent successful permanent HBP implantation. Three‐dimensional electro‐anatomical mapping system was used to facilitate the procedure and reduce the fluoroscopy time. There were no acute procedure‐related complications, and electrical parameters were stable at short‐term follow‐up.