A finely tuned interplay between calcium binding, ionic strength and pH modulates conformational and oligomerization equilibria in the Respiratory Syncytial Virus Matrix (M) protein.

As in most enveloped RNA viruses, the Respiratory Syncytial Virus Matrix (RSV-M) protein plays key roles in viral assembly and uncoating. It also plays non-structural roles related to transcription modulation through nucleo-cytoplasmic shuttling and nucleic acid binding ability. We dissected the structural and conformational changes underlying the switch between multiple functionalities, identifying Ca2+ binding as a key factor. To this end, we tackled the analysis of M's conformational stability and equilibria. While in silico calculations predict two potential calcium binding sites per protomer, purified RSV-M dimer contains only one strongly bound calcium ion per protomer. Incubation of RSV-M in the presence of excess Ca2+ leads to an increase in the thermal stability, confirming additional Ca2+ binding sites. Moreover, mild denaturant concentrations trigger the formation of higher order oligomers which are otherwise prevented under Ca2+ saturation conditions, in line with the stabilizing effect of the additional low affinity binding site. On the other hand, Ca2+ removal by chelation at pH 7.0 causes a substantial decrease in the thermal stability leading to the formation of amorphous, spherical-like aggregates, as assessed by TEM. Even though the Ca2+ content modulates RSV-M oligomerization propensity, it does affect its weak RNA binding ability. RSV-M undergoes a substantial conformational change at pHs 4.0 to 5.0 that results in the exposure of hydrophobic surfaces, an increase beta sheet content but burial of tryptophan residues. While low ionic strength promotes dimer dissociation at pH 4.0, physiological concentrations of NaCl lead to the formation of soluble oligomers smaller than 400 kDa at pH 4.0 or insoluble aggregates with tubular morphology at pH 5.0, supporting a fine tuning by pH. Furthermore, the dissociation constants estimated for the low- and high affinity calcium binding sites are 13 µM and 58 nM, respectively, suggesting an intracellular calcium sensing mechanism of RSV-M upon infection. We uncover a finely tuned interplay between calcium binding, ionic strength, and pH changes compatible with the different cellular compartments where M plays key roles, revealing diverse conformational equilibria, oligomerization, and high order structures, required to stabilize the virion particle by a layer of molecules positioned between the membrane and the nucleocapsid.

A conformational switch balances viral RNA accessibility and protection in a nucleocapsid ring model.

Virus from the Mononegavirales order share common features ranging from virion structure arrangement to mechanisms of replication and transcription. One of them is the way the nucleoprotein (N) wraps and protects the RNA genome from degradation by forming a highly ordered helical nucleocapsid. However, crystal structures from numerous Mononegavirales reveal that binding to the nucleoprotein results in occluded nucleotides that hinder base pairing necessary for transcription and replication. This hints at the existence of alternative conformations of the N protein that would impact on the protein-RNA interface, allowing for transient exposure of the nucleotides without complete RNA release. Moreover, the regulation between the alternative conformations should be finely tuned. Recombinant expression of N from the respiratory syncytial virus form regular N/RNA common among all Mononegavirales, and these constitute an ideal minimal unit for investigating the mechanisms through which these structures protect RNA so efficiently while allowing for partial accessibility during transcription and replication. Neither pH nor high ionic strength could dissociate the RNA but led to irreversible aggregation of the nucleoprotein. Low concentrations of guanidine chloride dissociated the RNA moiety but leading to irreversible aggregation of the protein moiety. On the other hand, high concentrations of urea and long incubation periods were required to remove bound RNA. Both denaturants eventually led to unfolding but converged in the formation of an RNA-free ß-enriched intermediate species that remained decameric even at high denaturant concentrations. Although the N-RNA rings interact with the phosphoprotein P, the scaffold of the RNA polymerase complex, this interaction did not lead to RNA dissociation from the rings in vitro. Thus, we have uncovered complex equilibria involving changes in secondary structure of N and RNA loosening, processes that must take place in the context of RNA transcription and replication, whose detailed mechanisms and cellular and viral participants need to be established.

Molecular and FISH analyses of a 53-kbp intact DNA fragment inserted by biolistics in wheat (Triticum aestivum L.) genome.

KEY MESSAGE: A large, 53-kbp, intact DNA fragment was inserted into the wheat ( Triticum aestivum L.) genome. FISH analyses of individual transgenic events revealed multiple insertions of intact fragments. Transferring large intact DNA fragments containing clusters of resistance genes or complete metabolic pathways into the wheat genome remains a challenge. In a previous work, we showed that the use of dephosphorylated cassettes for wheat transformation enabled the production of simple integration patterns. Here, we used the same technology to produce a cassette containing a 44-kb Arabidopsis thaliana BAC, flanked by one selection gene and one reporter gene. This 53-kb linear cassette was integrated in the bread wheat (Triticum aestivum L.) genome by biolistic transformation. Our results showed that transgenic plants harboring the entire cassette were generated. The inheritability of the cassette was demonstrated in the T1 and T2 generation. Surprisingly, FISH analysis performed on T1 progeny of independent events identified double genomic insertions of intact fragments in non-homoeologous positions. Inheritability of these double insertions was demonstrated by FISH analysis of the T1 generation. Relative conclusions that can be drawn from molecular or FISH analysis are discussed along with future prospects of the engineering of large fragments for wheat transformation or genome editing.

Structured surface reflector design for oblique incidence beam splitter at 610 GHz.

An iterative alternate projection-based algorithm is developed to design structured surface reflectors to operate as beam splitters at GHz and THz frequencies. To validate the method, a surface profile is determined to achieve a reflector at 610 GHz that generates four equal-intensity beams towards desired directions of ±12.6° with respect to the specular reflection axis. A prototype is fabricated and the beam splitter behavior is experimentally demonstrated. Measurements confirm a good agreement (within 1%) with computer simulations using Feko, validating the method. The beam splitter at 610 GHz has a measured efficiency of 78% under oblique incidence illumination that ensures a similar intensity between the four reflected beams (variation of about 1%).

The genome of Laccaria bicolor provides insights into mycorrhizal symbiosis.

Mycorrhizal symbioses--the union of roots and soil fungi--are universal in terrestrial ecosystems and may have been fundamental to land colonization by plants. Boreal, temperate and montane forests all depend on ectomycorrhizae. Identification of the primary factors that regulate symbiotic development and metabolic activity will therefore open the door to understanding the role of ectomycorrhizae in plant development and physiology, allowing the full ecological significance of this symbiosis to be explored. Here we report the genome sequence of the ectomycorrhizal basidiomycete Laccaria bicolor (Fig. 1) and highlight gene sets involved in rhizosphere colonization and symbiosis. This 65-megabase genome assembly contains approximately 20,000 predicted protein-encoding genes and a very large number of transposons and repeated sequences. We detected unexpected genomic features, most notably a battery of effector-type small secreted proteins (SSPs) with unknown function, several of which are only expressed in symbiotic tissues. The most highly expressed SSP accumulates in the proliferating hyphae colonizing the host root. The ectomycorrhizae-specific SSPs probably have a decisive role in the establishment of the symbiosis. The unexpected observation that the genome of L. bicolor lacks carbohydrate-active enzymes involved in degradation of plant cell walls, but maintains the ability to degrade non-plant cell wall polysaccharides, reveals the dual saprotrophic and biotrophic lifestyle of the mycorrhizal fungus that enables it to grow within both soil and living plant roots. The predicted gene inventory of the L. bicolor genome, therefore, points to previously unknown mechanisms of symbiosis operating in biotrophic mycorrhizal fungi. The availability of this genome provides an unparalleled opportunity to develop a deeper understanding of the processes by which symbionts interact with plants within their ecosystem to perform vital functions in the carbon and nitrogen cycles that are fundamental to sustainable plant productivity.

Maternal-infant transfer of SARS-CoV-2 antibodies following vaccination in pregnancy: A prospective cohort study.

OBJECTIVES: To measure and evaluate the impact of receiving severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines in pregnancy on immunoglobulin G (IgG) and immunoglobulin A (IgA) titres in maternal and infant samples. DESIGN: Prospective cohort study. SETTING: Tertiary obstetric centre. POPULATION OR SAMPLE: 52 pregnant women who received one or more SARS-CoV-2 vaccine doses during pregnancy and their neonates. METHODS: IgG and IgA concentrations against SARS-CoV-2 antigens were measured from samples collected at delivery and 4-6 weeks postpartum and compared using Spearman correlations. MAIN OUTCOME MEASURES: Maternal and infant IgG and IgA titres in response to vaccination and infection in pregnancy. RESULTS: In maternal serum collected at delivery, participants without evidence of prior infection who received 3 + doses of a SARS-CoV-2 vaccine had higher Anti-Spike (S) IgG geometric mean concentrations (log10 AU/mL)(GMC) than those who received 2 doses (3 + Doses = 5.00, 2 Doses = 4.60, p = 0.08). The differences in IgG Anti-S GMC were statistically significant in cord serum, and in postpartum samples of maternal serum, infant serum and breast milk (Cord GMCs: 3 + Doses = 5.32, 2 Doses = 4.98, p < 0.05; Postpartum maternal serum GMCs: 3 + Doses = 5.25, 2 Doses = 4.57, p < 0.001; Postpartum infant serum GMCs: 3 + Doses = 5.10, 2 Doses = 4.72, p = 0.03; Postpartum breast milk GMCs: 3 + Doses = 2.61, 2 Doses = 1.94, p < 0.0001). Among participants with 3 + Doses, those with evidence of SARS-CoV-2 infection had statistically significant higher anti-S IgG GMCs than those without prior infection (Maternal serum at delivery: SARS-CoV-2+=5.65, SARS-CoV-2-=5.00, p = 0.004; Cord: SARS-CoV-2+=5.68, SARS-CoV-2-=5.32, p = 0.02; Postpartum maternal serum: SARS-CoV-2+=5.66, SARS-CoV-2-=5.25, p < 0.001; postpartum infant serum: SARS-CoV-2+=5.50, SARS-CoV-2-=5.10, p = 0.003; Postpartum breast milk: SARS-COV-2+=3.25, SARS-COV-2-=2.61, p = 0.009). Significant positive correlations were found for anti-S IgG titres between paired maternal and infant samples at delivery and postpartum (Delivery: R = 0.91, p < 0.001; postpartum: R = 0.86, p < 0.001). CONCLUSIONS: Receipt of a SARS-CoV-2 vaccine and SARS-CoV-2 infection elicit strong IgG and IgA antibody responses in pregnant women with evidence of transplacental transfer to the fetus.

[Endoscopic submucosal dissection]. / Endoskopische Submukosadissektion.

Endoscopic submucosal dissection (ESD) was developed in Japan but has now also become permanently established in various centers in Europe. ESD is an endoscopic en bloc mucosal resection technique for the treatment of early cancers with a diameter >1 cm and also superficial precancerous lesions, which could only be removed unsatisfactorily in several fragments or with uncertain lateral safety margins using previous loop excision procedures. Using ESD a lesion is excised after circular marking and generous submucosal injection with a safety margin of approximately 5 mm and subsequently resected at the level of the submucosa with a 1-3 mm short diathermic knife. ESD requires high technical skills in interventional endoscopy and is more time-consuming than snare resection techniques. However, numerous studies have shown a clear superiority for ESD with respect to the R0 resection rate and the local recurrence rate. The present article gives a current review of the use of ESD in the upper and lower gastrointestinal tract and demonstrates perspectives of the procedure.

Reactogenicity, pregnancy outcomes, and SARS-CoV-2 infection following COVID-19 vaccination during pregnancy in Canada: A national prospective cohort study.

OBJECTIVE: To describe self-reported reactogenicity, pregnancy outcomes, and SARS-CoV-2 infection following COVID-19 vaccination during pregnancy. DESIGN: National, prospective cohort study. SETTING: Participants across Canada were enrolled from July 2021 until June 2022. POPULATION: Individuals pregnant during the COVID-19 pandemic, regardless of vaccination status, were included. METHODS: The Canadian COVID-19 Vaccine Registry for Pregnant and Lactating Individuals (COVERED) was advertised through traditional and social media. Surveys were administered at baseline, following each vaccine dose if vaccinated, pregnancy conclusion, and every two months for 14 months. Changes to pregnancy or vaccination status, SARS-CoV-2 infections, or significant health events were recorded. MAIN OUTCOME MEASURES: Reactogenicity (local and systemic adverse events, and serious adverse events) within 1 week post-vaccination, pregnancy and neonatal outcomes, and subsequent SARS-CoV-2 infection. RESULTS: Among 2868 participants who received 1-2 doses of a COVID-19 vaccine during pregnancy, adverse events described included: headache (19.5-33.9%), nausea (4.8-13.8%), fever (2.7-10.2%), and myalgia (33.4-42.2%). Reactogenicity was highest after the 2nd dose of vaccine in pregnancy. Compared to 1660 unvaccinated participants, there were no statistically significant differences in adverse pregnancy or infant outcomes, aside from an increased risk of NICU admission ≥ 24 h among the unvaccinated group. During follow-up, there was a higher rate of participant-reported SARS-CoV-2 infection in the unvaccinated compared to the vaccinated group (18[47.4%] vs. 786[27.3%]). CONCLUSIONS: Participant-reported reactogenicity was similar to reports from non-pregnant adults. There was no increase in adverse pregnancy and birth outcomes among vaccinated vs. unvaccinated participants and lower rates of SARS-CoV-2 infection were reported in vaccinated participants. TWEETABLE ABSTRACT: No significant increase in adverse pregnancy or infant outcomes among vaccinated versus unvaccinated pregnant women in Canada.

Colon capsule endoscopy: European Society of Gastrointestinal Endoscopy (ESGE) Guideline.

PillCam colon capsule endoscopy (CCE) is an innovative noninvasive, and painless ingestible capsule technique that allows exploration of the colon without the need for sedation and gas insufflation. Although it is already available in European and other countries, the clinical indications for CCE as well as the reporting and work-up of detected findings have not yet been standardized. The aim of this evidence-based and consensus-based guideline, commissioned by the European Society of Gastrointestinal Endoscopy (ESGE) is to furnish healthcare providers with a comprehensive framework for potential implementation of this technique in a clinical setting.

International Conference on Capsule and Double-Balloon Endoscopy (ICCD). Paris, 27-28 August 2010.

This review reports the highlights of the first International Conference on Capsule and Double-Balloon Endoscopy, held in Paris (27 - 28 August 2010), under the chairmanship of Professor G. Gay (Nancy, France). The conference was devoted to the technical issues and clinical experience related to capsule endoscopy and double-balloon endoscopy (DBE), techniques that have been developed over the past decade for the endoscopic investigation of intestinal diseases. The conference focused on the technical advances of new-generation endoscopes and capsules and the clinical experience gained with capsule endoscopy and DBE. Important advances in the knowledge of obscure gastrointestinal bleeding, inflammatory bowel diseases, and celiac disease were also discussed. The second part of the conference was mainly devoted to the investigation of the colon and discussed the technical issues and results of initial controlled studies of the colonic capsule vs. conventional colonoscopy. In addition, the clinical usefulness of double colonoscopy was demonstrated in patients with previously failed colonoscopy. Finally, the role of DBE was discussed with respect to the investigation and treatment of patients with surgically modified anatomy. The conference was attended by 750 delegates and presented a unique opportunity for an in-depth review of the current knowledge in the field and to ascertain/determine the main lines for the future developments of capsule endoscopy and DBE.

CMOS compatible strategy based on selective atomic layer deposition of a hard mask for transferring block copolymer lithography patterns.

A generic, CMOS compatible strategy for transferring a block copolymer template to a semiconductor substrate is demonstrated. An aluminum oxide (Al(2)O(3)) hard mask is selectively deposited by atomic layer deposition in an organized array of holes obtained in a PS matrix via PS-b-PMMA self-assembly. The Al(2)O(3) nanodots act as a highly resistant mask to plasma etching, and are used to pattern high aspect ratio (>10) silicon nanowires and nanopillars.

Different patterns of regulation for the copper and cadmium metallothioneins of the ectomycorrhizal fungus Hebeloma cylindrosporum.

Metallothioneins (MTs) are small cysteine-rich peptides involved in metal homeostasis and detoxification. We have characterized two MT genes, HcMT1 and HcMT2, from the ectomycorrhizal fungus Hebeloma cylindrosporum in this study. Expression of HcMT1 and HcMT2 in H. cylindrosporum under metal stress conditions was studied by competitive reverse transcription-PCR analysis. The full-length cDNAs were used to perform functional complementation in mutant strains of Saccharomyces cerevisiae. As revealed by heterologous complementation assays in yeast, HcMT1 and HcMT2 each encode a functional polypeptide capable of conferring increased tolerance against Cd and Cu, respectively. The expression levels of HcMT1 were observed to be at their maximum at 24 h, and they increased as a function of Cu concentration. HcMT2 was also induced by Cu, but the expression levels were lower than those for HcMT1. The mRNA accumulation of HcMT1 was not influenced by Cd, whereas Cd induced the transcription of HcMT2. Zn, Pb, and Ni did not affect the transcription of HcMT1 or of HcMT2. Southern blot analysis revealed that both of these genes are present as a single copy in H. cylindrosporum. While the promoters of both HcMT1 and HcMT2 contained the standard stress response elements implicated in the metal response, the numbers and varieties of potential regulatory elements were different in these promoters. These results show that ectomycorrhizal fungi encode different MTs and that each of them has a particular pattern of expression, suggesting that they play critical specific roles in improving the survival and growth of ectomycorrhizal trees in ecosystems contaminated by heavy metals.

Absence of microsomal triglyceride transfer protein in individuals with abetalipoproteinemia.

Abetalipoproteinemia is a human genetic disease that is characterized by a defect in the assembly or secretion of plasma very low density lipoproteins and chylomicrons. The microsomal triglyceride transfer protein (MTP), which is located in the lumen of microsomes isolated from the liver and intestine, has been proposed to function in lipoprotein assembly. MTP activity and the 88-kilodalton component of MTP were present in intestinal biopsy samples from eight control individuals but were absent in four abetalipoproteinemic subjects. This finding suggests that a defect in MTP is the basis for abetalipoproteinemia and that MTP is indeed required for lipoprotein assembly.

Short communication: The effect of centrifugation and resuspension on the recovery of Mycoplasma species from milk.

Low sensitivity of a single bulk tank milk culture is a major limitation for detection of mycoplasma organisms. We hypothesized that sedimentation of Mycoplasma spp. in a milk sample by centrifugation followed by resuspension in a small volume of fluid before agar plating would increase the ability to detect Mycoplasma spp. compared with direct conventional culture. The experiment was conducted to determine recovery of Mycoplasma spp. from milk as affected by 1) treatment (centrifugation vs. conventional method); 2) 2 species (Mycoplasma bovis and Mycoplasma californicum and 4 strains for each species); and 3) 4 different concentrations of Mycoplasma spp. (1,000, 100, 10, and 1 cfu/mL). A 5-mL portion of mycoplasma suspension from each strain was inoculated into 45 mL of fresh bulk tank milk to achieve concentrations of 1,000, 100, 10, and 1 cfu/mL. Treatment samples were vigorously mixed and centrifuged at 5,000 x g for 30 min. Control samples were vigorously mixed. All samples were plated on modified Hayflick agar. Plates were incubated at 37 degrees C and 5% CO(2) for 5 d. Mean (+/-SE) log(10) mycoplasma counts (cfu/mL) in the treatment groups (1.91 +/- 0.15) were higher than those in the control groups (1.70 +/- 0.16). Recovery of at least 1 mycoplasma colony on agar culture was 100% in both treatment and control groups at high, medium, and low concentrations. At the lowest concentration, recovery of at least 1 mycoplasma colony on agar culture in treatment and control groups was 75% (n = 12/16) and 18.75% (n = 3/16), respectively. Centrifugation of milk followed by suspension in a smaller volume of saline before conventional culture increased the ability to detect mycoplasma microorganisms in the milk sample compared with controls. Recovery by centrifugation appeared best at the lowest concentration where detection of a positive sample was 4 times more likely than when conventional methods were used.

Esophageal capsule endoscopy in patients with suspected esophageal disease: double blinded comparison with esophagogastroduodenoscopy and assessment of interobserver variability.

BACKGROUND AND STUDY AIMS: The study aim was to compare the diagnostic yield of capsule endoscopy and esophagogastroduodenoscopy (EGD) in patients with suspected esophageal disease. Secondary aims were to assess interobserver variability of capsule endoscopy readings and safety. PATIENTS AND METHODS: In total, 98 patients (53 men, 53 +/- 13 years) with an indication for EGD were included. The patient population was artificially enriched to include two thirds of patients with abnormal esophageal findings at EGD, which was followed by capsule endoscopy. Capsule recordings were blindly read by three endoscopists, one from the center that recruited the patient and two from the other center. Study outcomes were the findings described on EGD and capsule endoscopy, agreement between EGD and capsule endoscopy for findings, quality of the capsule recording, and interobserver agreement for capsule endoscopy quality and findings. RESULTS: EGD was normal in 34 patients and showed esophageal findings in 62 (esophagitis 28, hiatus hernia 21, varices 21, Barrett's esophagus 11, others 7). Average esophageal transit time of the capsule was 361 +/- 393 seconds. Capsule endoscopy was normal in 36 patients but detected esophagitis in 23, hiatus hernia in 0, varices in 23, Barrett's esophagus in 18, and others in 4. The positive predictive value of capsule endoscopy was 80.0 % and the negative predictive value was 61.1 %. Overall agreement per patient was moderate between EGD and capsule endoscopy for the per-patient (kappa = 0.42) and per-findings (kappa = 0.40) analyses. Interobserver agreement between capsule endoscopy readings was moderate for findings (kappa = 0.39) and quality assessment (kappa = 0.24). No adverse event was observed after either EGD or capsule endoscopy. CONCLUSION: In this study, despite artificial prevalence enrichment, capsule endoscopy showed a moderate sensitivity and specificity in the detection of esophageal diseases.

Molecular dynamics of the DNA-binding domain of the papillomavirus E2 transcriptional regulator uncover differential properties for DNA target accommodation.

Papillomaviruses are small DNA tumor viruses that infect mammalian hosts, with consequences from benign to cancerous lesions. The Early protein 2 is the master regulator for the virus life cycle, participating in gene transcription, DNA replication, and viral episome migration. All of these functions rely on primary target recognition by its dimeric DNA-binding domain. In this work, we performed molecular dynamics simulations in order to gain insights into the structural dynamics of the DNA-binding domains of two prototypic strains, human papillomavirus strain 16 and the bovine papillomavirus strain 1. The simulations underline different dynamic features in the two proteins. The human papillomavirus strain 16 domain displays a higher flexibility of the beta2-beta3 connecting loop in comparison with the bovine papillomavirus strain 1 domain, with a consequent effect on the DNA-binding helices, and thus on the modulation of DNA recognition. A compact beta-barrel is found in human papillomavirus strain 16, whereas the bovine papillomavirus strain 1 protein is characterized by a loose beta-barrel with a large number of cavities filled by water, which provides great flexibility. The rigidity of the human papillomavirus strain 16 beta-barrel prevents protein deformation, and, as a consequence, deformable spacers are the preferred targets in complex formation. In contrast, in bovine papillomavirus strain 1, a more deformable beta-barrel confers greater adaptability to the protein, allowing the binding of less flexible DNA regions. The flexibility data are confirmed by the experimental NMR S2 values, which are reproduced well by calculation. This feature may provide the protein with an ability to discriminate between spacer sequences. Clearly, the deformability required for the formation of the Early protein 2 C-terminal DNA-binding domain-DNA complexes of various types is based not only on the rigidity of the base sequences in the DNA spacers, but also on the intrinsic deformability properties of each domain.

Surface-wave interferometry on single subwavelength slit-groove structures fabricated on gold films.

We apply the technique of far-field interferometry to measure the properties of surface waves generated by two-dimensional (2D) single subwavelength slit-groove structures on gold films. The effective surface index of refraction n(surf) measured for the surface wave propagating over a distance of more than 12 mum is determined to be n(surf) = 1.016+/-0.004, to within experimental uncertainty close to the expected bound surface plasmon-polariton (SPP) value for a Au/Air interface of n (spp) = 1.018. We compare these measurements to finite-difference-time-domain (FDTD) numerical simulations of the optical field transmission through these devices. We find excellent agreement between the measurements and the simulations for n(surf). The measurements also show that the surface wave propagation parameter k(surf) exhibits transient behavior close to the slit, evolving smoothly from greater values asymptotically toward k (spp) over the first 2-3 mum of slit-groove distance x(sg). This behavior is confirmed by the FDTD simulations.

Double-balloon colonoscopy after failed conventional colonoscopy: a pilot series with a new instrument.

BACKGROUND AND STUDY AIMS: The endoscopes that were developed for double-balloon enteroscopy have been successfully used in cases of failed colonoscopy. This study was a pilot series in which a new colonoscope was tested that utilized this double-balloon principle. PATIENTS AND METHODS: A total of 29 patients (5 men, 24 women; mean age 54 years) in whom conventional colonoscopy had failed were included in this study. Both the failed colonoscopy and the double-balloon colonoscopy procedures were performed under general anesthesia, the usual practice in France. A prototype instrument (working length 152 cm, diameter 9.4 mm) designed to incorporate the principles of double-balloon enteroscopy was used. The completeness of colonoscopy was assessed according to conventional criteria by the achievement of a stable position in the cecum. The indicatons for the procedure, the time to reach the cecum, the need for fluoroscopic control, and adverse events were recorded. RESULTS: The previous colonoscopy failed due adhesions (n = 16), or to long or fixed loops (n = 13). Complete colonoscopy using the balloon method was achieved in 28/29 patients, taking an average time of 18 +/- 14 minutes; a long sigmoid loop limited the examination to the left flexure in one patient. Balloon colonoscopy using double-balloon methodology was used in 24 patients and the instrument was used without an overtube (i. e. using a single-balloon technique) in five patients. Fluoroscopy was used in 16 patients to monitor endoscope progression. No complications were reported. CONCLUSIONS: Double-balloon colonoscopy enables full colonic examination in almost all patients with a previous incomplete colonoscopy. The overtube should be used in most cases. The use of fluoroscopic assessment of scope progression could be reduced further with increasing experience.

Surface quality and surface waves on subwavelength-structured silver films.

We analyze the physical-chemical surface properties of single-slit, single-groove subwavelength-structured silver films with high-resolution transmission electron microscopy and calculate exact solutions to Maxwell's equations corresponding to recent far-field interferometry experiments using these structures. Contrary to a recent suggestion the surface analysis shows that the silver films are free of detectable contaminants. The finite-difference time-domain calculations, in excellent agreement with experiment, show a rapid fringe amplitude decrease in the near zone (slit-groove distance out to 3-4 wavelengths). Extrapolation to slit-groove distances beyond the near zone shows that the surface wave evolves to the expected bound surface plasmon polariton (SPP). Fourier analysis of these results indicates the presence of a distribution of transient, evanescent modes around the SPP that dephase and dissipate as the surface wave evolves from the near to the far zone.

Trophic relationships in an Arctic food web and implications for trace metal transfer.

Tissues of subsistence-harvested Arctic mammals were analyzed for silver (Ag), cadmium (Cd), and total mercury (THg). Muscle (or total body homogenates of potential fish and invertebrate prey) was analyzed for stable carbon (delta13C) and nitrogen (delta15N) isotopes to establish trophic interactions within the Arctic food chain. Food web magnification factors (FWMFs) and biomagnification factors for selected predator-prey scenarios (BMFs) were calculated to describe pathways of heavy metals in the Alaskan Arctic. FWMFs in this study indicate that magnification of selected heavy metals in the Arctic food web is not significant. Biomagnification of Cd occurs mainly in kidneys; calculated BMFs are higher for hepatic THg than renal THg for all predator-prey scenarios with the exception of polar bears (Ursus maritimus). In bears, the accumulation of renal THg is approximately 6 times higher than in liver. Magnification of hepatic Ag is minimal for all selected predator-prey scenarios. Though polar bears occupy a higher trophic level than belugas (Delphinapterus leucas), based on delta15N, the metal concentrations are either not statistically different between the two species or lower for bears. Similarly, concentrations of renal and hepatic Cd are significantly lower or not statistically different in polar bears compared to ringed (Phoca hispida) and bearded seals (Erignathus barbatus), their primary prey. THg, on the other hand, increased significantly from seal to polar bear tissues. Mean delta15N was lowest in muscle of Arctic fox (Alopex lagopus) and foxes also show the lowest levels of Hg, Cd and Ag in liver and kidney compared to the other species analyzed. These values are in good agreement with a diet dominated by terrestrial prey. Metal deposition in animal tissues is strongly dependent on biological factors such as diet, age, sex, body condition and health, and caution should be taken when interpreting magnification of dynamic and actively regulated trace metals.