Ultrathin Alvarez lens system actuated by artificial muscles.

A key feature of Alvarez lenses is that they may be tuned in focal length using lateral rather than axial translation, thus reducing the overall length of a focus-tunable optical system. Nevertheless the bulk of classical microsystems actuators limits further miniaturization. We present here a new, ultrathin focus-tunable Alvarez lens fabricated using molding techniques and actuated using liquid crystal elastomer (LCE) artificial muscle actuators. The large deformation generated by the LCE actuators permits the integration of the actuators in-plane with the mechanical and optical system and thus reduces the device thickness to only 1.6 mm. Movement of the Alvarez lens pair of 178 µm results in a focal length change of 3.3 mm, based on an initial focal length of 28.4 mm. This design is of considerable interest for realization of ultraflat focus-tunable and zoom systems.

Bimodal endoscopic probe combining white-light microscopy and optical coherence tomography.

We present a novel bimodal endoscopic imaging probe that can simultaneously provide full-field white-light video microscopy and confocal optical coherence tomography (OCT) depth scans. The two modalities rely on spectrally separated optical paths that run partially in parallel through a micro-optical bench system, which has a cross-section of only 2 mm×2.76 mm and is realized via standard silicon micromachining techniques. With a numerical aperture of 0.061, the video modality has a resolution and field of view of 9.3 and 1240 µm×1080 µm, respectively. The resolution is limited by the pixel spacing of the coherent fiber bundle, which relays the acquired image from the distal to the proximal end. A custom-designed diffractive optical element placed within the video imaging path significantly improves the image contrast by up to 45% in the medium frequency range. The OCT modality is optimized for 830 nm center wavelength, and works in a confocal arrangement with an NA of 0.018. It provides single-point depth probing at the center of the video image with a lateral resolution of 20 µm. Through its compact footprint and enhanced functionality, the probe can provide depth-resolved guiding capability for existing laparoscopes and represents a major step toward a new class of multimodal endoscopic imaging probes.

Jet printing of convex and concave polymer micro-lenses.

We describe a novel approach for fabricating customized convex as well as concave micro-lenses using substrates with sophisticated pinning architecture and utilizing a drop-on-demand jet printer. The polymeric lens material deposited on the wafer is cured by UV light irradiation yielding lenses with high quality surfaces. Surface shape and roughness of the cured polymer lenses are characterized by white light interferometry. Their optical quality is demonstrated by imaging an USAF1951 test chart. The evaluated modulation transfer function is compared to Zemax simulations as a benchmark for the fabricated lenses.

[Enhanced imaging in urological endoscopy]. / Erweiterte Bildgebung in der urologischen Endoskopie.

White light cystoscopy and the concise documentation of pathological findings are standard diagnostic procedures in urology. Additional imaging modalities and technical innovations may support clinicians in the detection of bladder tumors. Modern endoscopy systems provide ultra-high-resolution imaging and the option of digital contrast enhancement. Photodynamic diagnostics and narrow band imaging are well-established in clinical routine and have shown significant benefits in the detection of bladder cancer. By means of multispectral imaging, different modalities can now be combined in real-time. Probe-based procedures such as optical coherence tomography (OCT) or Raman spectroscopy can further contribute to advanced imaging through an "optical biopsy" which may primarily improve diagnostics in the upper urinary tract. The aim of all techniques is to optimize the detection rate in order to achieve a more accurate diagnosis, resection and lower recurrence rates. Current research projects aim to digitalize the documentation of endoscopy and also make it more patient- and user-friendly. In the future, the use of image processing and artificial intelligence may automatically support the surgeon during endoscopy.

PEGylation of cyanovirin-N, an entry inhibitor of HIV.

Cyanovirin-N (CV-N) is a potent inhibitor of human immunodeficiency virus and many other viruses. It has a high potential for use as a systemic compound to control viral load or in the development of microbicides to prevent primary viral infection. Due to its cyanobacterial origin it is likely to show the typical drawbacks associated with pharmaceutical use of foreign proteins such as short plasma half-life, proteolysis and immunogenicity. Several strategies were used to covalently bond poly(ethylene glycol) (PEGylate) to CV-N. Random PEGylation at lysine residues resulted in poor retention of antiviral activity. Many site-directed mutants were made to test site-specific PEGylation. One mutant, where glutamine 62 was replaced with cysteine (CV-N(Q62C)) and PEGylated with maleimide activated PEG, retained significant anti-HIV activity in vitro.

X-ray diffraction study of fast and slow mammalian skeletal muscle in the live relaxed state.

Experimental methods have been developed by which fast and slow muscles of the mouse can be maintained in good condition for the time needed to record detailed X-ray patterns. The results presented here show that thick filaments of both types of fibre show only minor differences from those of frog muscle. However, in slow muscle alone a remarkable transformation of this structure can occur.

The Mycobacterium tuberculosis mysB gene product is a functional equivalent of the Escherichia coli sigma factor, KatF.

Mycobacterium tuberculosis, the causative agent of tuberculosis, may remain dormant within its host for many years. The nature of this dormant or latent state is not known, but it may be a specialized form of the stationary growth phase. In Escherichia coli, KatF (or RpoS) is the major stationary phase sigma factor regulating an array of genes expressed in this phase of growth. A potential M. tuberculosis katF homologue was cloned using a fragment of the E. coli katF gene as a probe. DNA sequence analysis of a resultant clone showed 100% identity to a fragment of DNA encoding the M. tuberculosis mysA and mysB genes. Overexpression of mysB in M. bovis BCG resulted in an increase in katG mRNA and catalase and peroxidase activity, and an increase in sensitivity of the cells to isoniazid. An increase in katG promoter activity from a reporter vector was demonstrated when mysB was overexpressed from the same plasmid, indicating a direct relationship between MysB and katG expression.

Characterisation of a lipoprotein in Mycobacterium bovis (BCG) with sequence similarity to the secreted protein MPB70.

The monoclonal antibody, mAb3C4, raised against sonicated Mycobacterium bovis (Mb) BCG (Tokyo strain 172) cells recognises a 23-kDa protein in the cell wall. The gene encoding this protein was cloned and sequenced and found to be 100% homologous to mpb83 and mpt83 and the putative protein to have a 76% sequence similarity to the secreted, Mb-specific protein, MPB70. MPB83 contains the amino acid (aa) sequence LAGC, which corresponds to the consensus sequence for bacterial lipoprotein modification and processing. MPB83 associated with the detergent phase when separated with Triton X-114 confirming that it is a lipoprotein. When the putative site of acylation, the Cys in the sequence LAGC, was substituted with Ser, the mutated MPB83 associated with the aqueous phase. The cloned gene was used to determine the distribution of mpb83 in various Mycobacterium species. The gene was present in the M. tuberculosis (Mt) complex organisms, as well as in M. kansasii. In addition, Southern blot analysis of Mb and Mt DNA indicated that the mpb83 and mpb70 genes are located close to each other on the genome. Western blot analysis of cell lysates of various Mycobacterium species indicated that only Mt H37Rv and H37Ra produced proteins which reacted with mAb3C4. Furthermore, only two out of six of the Mb field isolates produced detectable antigen, indicating that expression of the mpb83 gene is variable within the Mt complex organisms.

Growth phase-associated changes in protein expression in Mycobacterium smegmatis identify a new low molecular weight heat shock protein.

De novo protein synthesis and the heat-shock response during different stages of bacterial culture of Mycobacterium smegmatis LR222 were investigated. A discontinuance in the increase in number of colony forming units occurred at mid-exponential phase of growth. This coincided with a plateau in the ATP content of the culture, a reduction in the synthesis of exponential phase proteins (58, 30.5, and 20 kDa), a transitory synthesis of a 32 kDa protein and the induction of stationary-phase proteins (48, 46, 31, 25, and 20 kDa). The response to heat shock showed a growth-phase dependency, with the highest fold-induction of the 75 kDa (DnaK) protein occurring during the transitory cessation in the increase in CFU, while the greatest increase of the 95 kDa, 66 kDa (GroEL), and approximately 17 kDa (a doublet) proteins occurred during stationary phase. The approximately 17 kDa doublet was resolved into four polypeptides by two-dimensional electrophoresis. Mass spectrometric analysis of the sequence of one polypeptide (named Hsp17-2, 16.8 kDa) revealed significant homology to a conserved, 16.2 kDa, hypothetical protein of unknown function in Mycobacterium tuberculosis H37Rv. The increased synthesis of Hsp17-2 in response to heat shock suggests that it may represent a new low molecular weight heat shock protein.

16S rRNA sequence analysis of an isolate of Mycobacterium haemophilum from a heart transplant patient.

Biopsy samples from a heart transplant patient with cellulitis and bursitis yielded an isolate of Mycobacterium haemophilum. The isolate was identified on the basis of a growth requirement for haemin or ferric ammonium citrate, growth at 30 degrees C but not at 37 degrees C, negative catalase test, intracellular growth in McCoy fibroblasts and sequence identify with a portion of the 16S rRNA sequence of the type strain. In comparisons with known 16S rRNA sequences, M. haemophilum grouped with other pathogenic, slow-growing mycobacteria, showing close sequence similarity to M. marinum (98.8%) and lower similarity to M. ulcerans and M. tuberculosis complex organisms. M. haemophilum and M. marium share other features including optimal growth at 30 degrees C and the ability to cause superficial skin lesions in man.

Efficacy of montelukast in the treatment of chronic obstructive pulmonary disease in five horses.

Five horses with moderate to severe chronic obstructive pulmonary disease (COPD) were treated with 0.11 (0.01) mg/kg bodyweight of montelukast, a cysteinyl leukotriene receptor antagonist, once a day for 26 days. The horses were evaluated clinically and endoscopically and subjected to arterial blood gas analysis and lung function tests before and after the period of treatment, and the plasma concentrations of montelukast were determined by high-performance liquid chromatography with fluorescence detection. The treatment did not result in statistically significant differences in the total scores of clinical and endoscopical signs, or in the difference in the arterioalveolar partial pressure of oxygen, maximal changes in pleural pressure, pulmonary resistance or dynamic compliance. The mean (sd) peak plasma concentration (C(max0) of montelukast was 12 (4) ng/ml and was reached 66 (13) minutes (t(max)) after its oral administration. The dose of montelukast per kg bodyweight was approximately the same as that for human beings, but the C(max) in the horses was 28 times lower and the t(max) was reached in one-fifth of the time, suggesting that its oral bioavailability may be lower.

Study of the effect of Bronchipret on the lung function of five Austrian saddle horses suffering recurrent airway obstruction (heaves).

The effects of an oral preparation containing an extract of thyme and primula (Bronchipret; Bionorica) on the lung function of five horses suffering heaves were determined in a longitudinal study. The horses accepted the product well. The plasma concentrations of the marker substance, thymol, indicated that at least one of the substances in the extract had been absorbed from the gastrointestinal tract. The compliance, pulmonary pressure and airway resistance of the horses' lungs were all significantly improved after one month of treatment However, the severity of their clinical signs and their arterial oxygen partial pressure had not improved significantly.

The use of (GTG)5 oligonucleotide as an RAPD primer to type Campylobacter concisus.

AIM: DNA fingerprinting using (GTG)(5) oligonucleotide as a primer in a random amplified polymorphic DNA (RAPD) assay was assessed by typing isolates of Campylobacter concisus strains, collected over a period of 8 years. METHODS AND RESULTS: RAPD analysis using the (GTG)(5) oligonucleotide as a primer was used to type 100 isolates of C. concisus comprising mostly isolates from children with diarrhoea. Using this method, 86% of the isolates were found to be genotypically diverse. Of these heterogeneous isolates, 25 of the strains were also shown to be genetically distinct in a previous study using pulsed field gel electrophoresis. The remaining isolates (14) could be classified into five profile groups based on the DNA fingerprinting patterns. The assay successfully identified epidemiologically linked strains from the unrelated genetically diverse pool of strains. CONCLUSIONS: Laboratory RADP typing using the (GTG)(5) primer proved to be useful in distinguishing related strains of C. concisus from a large pool of unrelated strains of this organism. SIGNIFICANCE AND IMPACT OF THE STUDY: RAPD typing using (GTG)(5) is a simple method that could be used to investigate the epidemiology of C. concisus. The results suggest that homologous lineages of C. concisus may exist within an otherwise heterogeneous species complex. However, these data need to be confirmed using a more robust typing method.

Expression of Bacillus amyloliquefaciens amylase and Vibrio alginolyticus protease A fusion genes.

Previously we reported [Deane, S. M., Maharaj, R., Robb, F. T. & Woods, D. R. (1987) Journal of General Microbiology 133, 2295-2302] that the production of a Vibrio alginolyticus SDS-resistant alkaline serine protease (Pro A) cloned in Escherichia coli was characterized by a 12 h delay between the synthesis of an inactive precursor and secretion of active Pro A. Replacement of the V. alginolyticus promoter region by the alpha-amylase promoter region from Bacillus amyloliquefaciens resulted in the simultaneous synthesis and secretion of Pro A in E. coli. The V. alginolyticus pro A gene cloned on a shuttle vector did not produce active Pro A in Bacillus subtilis. Although Pro A has a typical Gram-positive signal sequence, it was not functional in B. subtilis. Replacement of the Pro A signal sequence with the alpha-amylase signal sequence resulted in the production of active Pro A in B. subtilis.

A reporter gene vector to investigate the regulation of glutamine synthetase in Bacteroides fragilis Bf1.

The Clostridium acetobutylicum eglA gene, encoding a beta-1,4-endoglucanase (EG), was shown to be a useful reporter gene for the study of gene expression in Bacteroides fragilis. The eglA reporter gene has the advantages that it can be easily identified in both Escherichia coli and B. fragilis on agar media containing carboxymethylcellulose, and EG production can be rapidly quantified in liquid medium. Since the B. fragilis glutamine synthetase (GS) is inactivated in permeabilized cells and cell extracts, the eglA reporter gene was used to study the regulation of GS production in B. fragilis. Gene fusions containing the GS glnA promoter region fused to the promoterless eglA gene showed that glnA expression was regulated by nitrogen in B. fragilis at the transcriptional level. A glnA upstream region containing a near-perfect direct repeat sequence was essential for efficient GS expression and for regulation by nitrogen.

A survey on pain complaints and health care utilization in a German population sample.

A postal survey was carried out on every 71st person aged between 18 and 80 in the population registers in County Regierungsbezirk Karlsruhe in the State of Baden-Würtemberg. It asked 2127 persons whether they had, in the previous 6 months, experienced any form of unduly prolonged pain (as distinct from brief intercurrent self-limiting episodes related to injury inflammation etc.) and, if so, to specify its location, duration, severity and persistence. It also sought information on the resulting calls on healthcare professionals and the degree of satisfaction with treatments received. The age and gender distributions of the sample selected for survey matched those in the population from which it was drawn. Of the 1420 respondents, only 1304 declared their age and gender--a condition for inclusion in the analysis. Of these, 610 reported some form of unduly prolonged pain, which had lasted more than a year in 530. For all pain lasting longer than a year, the estimated prevalence of mild pain was 11%, severe 25% and intolerable 3.5%: the corresponding estimates for persistent as opposed to episodic pain were 2% for mild, 10% for sever and 1% for intolerable. Pain was present in more than one anatomical location in most of those who reported it. Musculoskeletal pain was overwhelmingly the most common. Increasing age, obesity and being female pre-disposed to the reporting of pain, with women being more liable to report headache and pain in the neck and shoulder. One hundred and thirty-six pain reporters either gave no information on consultation or sought no help from healthcare professionals: a third of the remainder consulted more than one professional, with general practitioners and specialists in physical medicine (niedergelassener Orthopäde) being the most common. A wide variety of treatments were used, with oral medications, massage, exercises, mud pack and heat treatment being the most popular; two-thirds of sufferers used more than one type of treatment. The most popular types of treatment tended also to be the most successful, except for oral medication (which was also the most heterogeneous). Multiple logistic regression analyses identified consistent associations between duration and severity of pain, the number of sites where it was reported, the numbers of healthcare professionals consulted and the number of treatments tried, and the same groupings of features were associated with decreased likelihood of overall satisfaction with treatment received.