Sub-100 nm focusing of short wavelength plasmons in homogeneous 2D space.

We present a direct measurement of short-wavelength plasmons focused into a sub-100 nm spot in homogeneous (translation invariant) 2D space. The short-wavelength (SW) surface plasmon polaritons (SPP) are achieved in metal-insulator-insulator (MII) platform consisting of silver, silicon nitride, and air. This platform is homogeneous in two spatial directions and supports SPP at wavelength more than two times shorter than that in free space yet interacts with the outer world through the evanescent tail in air. We use an apertureless (scattering) near-field scanning optical microscope (NSOM) to map directly the amplitude and phase of these SW-SPP and show they can be focused to under 70 nm without structurally assisted confinement such as nanoantennas or nanofocusing. This, along with the use of visible light at 532 nm which is suitable for optical microscopy, can open new directions in direct biological and medical imaging at the sub-100 nm resolution regime.

Effect of melatonin on sleep quality of COPD intensive care patients: a pilot study.

Sleep deprivation is extremely common in the intensive care unit (ICU), and this lack of sleep is associated with low melatonin secretion. The objective of the current study was to explore the effect of exogenous melatonin administration on sleep quality in patients hospitalized in the pulmonary intensive care unit (ICU). We performed a double-blind, placebo-controlled study in the pulmonary ICU of a tertiary care hospital. Eight adult patients hospitalized in the pulmonary ICU with respiratory failure caused by exacerbation of chronic obstructive pulmonary disease (COPD) or with pneumonia were studied. Patients received either 3 mg of controlled-release melatonin or a placebo at 22:00, and sleep quality was evaluated by wrist actigraphy. Treatment with controlled-release melatonin dramatically improved both the duration and quality of sleep in this group of patients. Our results suggest that melatonin administration to patients in intensive care units may be indicated as a treatment for sleep induction and resynchronization of the "biologic clock." This treatment may also help in the prevention of the "ICU syndrome" and accelerate the healing process.

Patients in the intensive care unit suffer from severe lack of sleep associated with loss of normal melatonin secretion pattern.

BACKGROUND: Patients hospitalized in the intensive care unit (ICU) tend to become agitated and confused, and many even develop temporary psychoses (the ICU syndrome). We wondered whether the regulation of sleep and the secretion of melatonin is abnormal in ICU patients. Therefore, we studied the association of sleep-wake pattern in patients hospitalized in the ICU, their melatonin secretion rates, and profile compared with a control group of patients in general medical wards. METHODS: Sleep was assessed by actigraphy. Urine was collected every 3 hours for 24 hours. Melatonin secretion was assessed by measuring the melatonin metabolite 6-sulphatoxymelatonin by enzyme-linked immunosorbent assay. RESULTS: Actigraphy suggested that the ICU patients lacked normal sleep behavior for the entire study period, except for occasional short naps. Compared with controls, the nocturnal peak of melatonin secretion was absent, except in two patients in the nonventilated group, and showed a flat curve. CONCLUSIONS: Our results suggest that lack of sleep is indeed a severe problem in ICU patients and is accompanied by impairment of normal melatonin secretion. The possibility that melatonin administration may prove useful in improving sleep patterns in ICU patients deserves further study.

Digoxinlike immunoreactive factor isolated from human pleural fluid is structurally similar to digoxin.

To further define the chemical structure of human endogenous digoxinlike immunoreactive factors (DLIF) we used human pleural effusions as a source of the substance. Digoxinlike immunoreactive factor activity was detected by radioimmunoassay in the pleural fluid of each of four patients; average concentration was 0.35 ng/mL. The chemical profile of DLIF was determined by initial extraction and concentration of DLIF by ion exchange chromatography followed by reverse phase-high-pressure liquid chromatography (RP-HPLC) separation and purification. Using high-pressure liquid chromatography cochromatography of DLIF, together with several radioactively marked glycosides, we observed a single peak of DLIF activity that was chromatographically identical to digoxin. The present study further supports the recent finding that DLIF is related structurally to the cardiac glycosides, and for the first time it has been proven that DLIF is present in pleural fluids.

Identification and preliminary characterization of two human digitalis-like substances that are structurally related to digoxin and ouabain.

In order to characterize the structure of endogenous digitalis-like immunoreactive factor (DLIF), we utilized peritoneal dialysis fluid from patients with chronic renal failure as a source of endogenous digitalis-like immunoreactive factor (DLIF), and subjected it to one-step ion exchange chromatography, followed by one step reverse HPLC. Crude dialysis fluid contained 0.09 ng/ml of DLIF, and using Amberlite XAD-2 chromatography we extracted 110 ng of DLIF from 800 ml of dialysis fluid. By applying this partially purified DLIF to our HPLC system, we discerned three peaks of DLIF activity, with retention times of 34, 58 and 63 minutes. The first peak overlapped the elution profile of ouabain, and the third peak co-eluted precisely with digoxin. The second DLIF peak was not in proximity to any of the digitalis-like markers employed. Thus, our results indicate that DLIF isolated from peritoneal dialysis fluid exists in three distinct forms, one of which resembles ouabain, and one which is identical to digoxin.

Endogenous digoxin-like immunoreactive factor is elevated in advanced chronic respiratory failure.

Digoxin-like immunoreactive factor (DLIF) is an endogenous substance with natriuretic and diuretic activity. Elevated plasma levels of DLIF are found in various clinical states characterized by water and sodium retention. Chronic respiratory failure, particularly of an advanced stage, also is frequently associated with water and sodium retention. In order to determine whether elevated plasma levels of DLIF are present in chronic respiratory failure, we measured plasma DLIF levels in seven patients (four with COPD [two of whom had associated sleep apnea disturbance] and three with kyphoscoliosis) suffering from advanced chronic respiratory failure with severe hypoxemia and hypercapnia. We found that in these patients plasma levels of DLIF were significantly higher than in healthy control subjects. We conclude that patients with advanced chronic respiratory failure respond with increased levels of DLIF. This may represent an attempt at homeostasis of water and sodium metabolism which is frequently deranged in this clinical condition.

Determinants of site-specific recombination in the lambdoid coliphage HK022. An evolutionary change in specificity.

The temperate bacteriophage HK022, like its relative lambda, inserts its chromosome into a specific site in the bacterial chromosome during lysogenization and excises it after induction. However, we find that the recombinational specificities of the two phages differ: they use different bacterial sites, and neither promotes efficient insertion or excision of the other phage chromosome. In order to determine the basis for this difference in specificity, we sequenced the HK022 elements that are involved in insertion and excision, and compared them to the corresponding lambda elements. The location, orientation, size and overall arrangement of the int and xis genes and the phage attachment sites are nearly identical in the two genomes, as is common for other functionally related elements in lambdoid phages. The Xis proteins of the two phages are functionally interchangeable, and their predicted amino acid sequences differ by but one residue. In contrast, the two Int proteins are not functionally interchangeable, and their sequences, although similar, differ at many positions. These sequence differences are not uniformly distributed: the amino-terminal 55 residues are completely conserved, but the remaining 302 show a pattern of differences interspersed with identities and conservative changes. These findings imply that the specificity difference between HK022 and lambda site-specific recombination is a consequence of the inability of the respective Int proteins to recognize pairs of heterologous attachment sites. The two phage attachment sites are remarkably similar, especially the two "arm" segments, which in lambda contain binding sites for Int, Xis and integration host factor. They are less similar in the segment between the two arms, which in lambda contains the points of recombinational strand exchange and a second class of binding site for Int protein (the "core-type" sites). The two bacterial attachment sites are quite different, although both have a short stretch of perfect homology with their respective phage partners at the points of strand exchange. We propose that the two Int proteins recognize similar or identical sites in the arms of their cognate attachment sites, and that differences in binding or action at the core-type sites is responsible for the divergent specificities. Genetic experiments and sequence comparisons suggest that both proteins recognize different but overlapping families of core-type sites, and that divergence in specificity has been achieved by an alternating succession of small, mutually compatible changes in protein and site.

Endogenous digoxin-like factor in neonates: effect of age and relation to serum bilirubin levels.

Endogenous digoxin-like immunoreactive factor(s) (DLIF) have been found in serum and urine of newborn infants, including those born prematurely. We assessed the effect of age on serum levels of DLIF in 73 samples obtained from 66 healthy full term newborn infants at birth and during the first two months of life. DLIF concentrations were highest at birth and fell progressively with age. In cord blood, DLIF levels were 0.73 +/- 0.35 ng/ml (mean +/- SD). DLIF concentrations were 0.45 +/- 0.11 ng/ml on day 1, 0.26 +/- 0.08 ng/ml on day 3, 0.19 +/- 0.07 ng/ml on day 5, 0.17 +/- 0.09 ng/ml on day 11, 0.11 +/- 0.02 ng/ml on days 15-30, and not detectable after 45 days of life. We also studied the relation between serum levels of DLIF and bilirubin in 23 jaundiced newborns between 3-5 days of life. We found a highly significant positive correlation between serum bilirubin concentrations and DLIF. These findings support the assumption that DLIF plays a role in impeding bilirubin excretion in the neonatal period, perhaps by inhibiting the activity of (Na-K)ATPase.

Digoxin-like immunoreactivity: occurrence in three molecular forms and partial characterization.

Digoxin-like immunoreactivity (DLI) has been found in serum from subjects with a variety of physiological and pathological conditions, including uremia, liver disease, pregnancy, and the neonatal period. The physicochemical nature of this material is still not known. Using gel filtration chromatography, extraction with methylene chloride, and treatment with beta-glucuronidase, we determined that serum DLI exists in three forms: protein-bound, glucuronidated, and free, with respective mol wt of 5000, 400, and 230. In serum, DLI exists in the protein-bound and free forms, while in urine, DLI is found in the glucuronidated and free states.

Fresh and cryopreserved fetal bones replacing massive bone loss in rats.

Cartilaginous fetal bones from rat preserved by deep freezing procedures were compared to comparable fresh bones with regard to the following parameters: chemical composition, water and uronic acid contents; cell viability measured by the rate of proteoglycan synthesis; mineralization-ossification status by calcium binding; matrix integrity by the release of uronic acid containing substances; and biological activity as transplants inducing the formation of bone. The transplanted material was chemically analyzed and checked for its rate of proteoglycan synthesis. The quality of the formed bone was similar whether isogeneic or allogeneic, fresh or cryopreserved bone was employed as transplant material. Evidently those various fetal bones may be of clinical value whenever the need for replacement of massive bone loss arises. Although the viability and the cartilaginous nature of the graft are critical, the isogeneity and freshness are of a quantitative advantage only. These biochemical observations were confirmed by roentgenological and histological evaluations of the grafts. An optimal cryopreserving procedure and tests for examining bone candidates for successful grafting are described.

Viral genome RNA serves as messenger early in the infectious cycle of murine leukemia virus.

When NIH/3T3 mouse fibroblasts were infected with the Moloney strain of murine leukemia virus, part of the viral genome RNA molecules were detected in polyribosomes of the infected cells early in the infectious cycle. The binding appears to be specific, since we could demonstrate the release of viral RNA from polyribosomes with EDTA. Moreover, when infection occurred in the presence of cycloheximide, most viral RNA molecules were detected in the free cytoplasm. Size analysis on polyribosomal viral RNA molecules indicated that two size class molecules, 38S and 23S, are present in polyribosomes at 3 h after infection. Analysis of the polyriboadenylate [poly(rA)] content of viral RNA extracted from infected polyribosomes demonstrated that such molecules bind with greatest abundance at 3 h after infection, as has been detected with total viral RNA. No molecules lacking poly(rA) stretches could be detected in polyribosomes. Furthermore, when a similar analysis was performed on unbound molecules present in the free cytoplasm, identical results were obtained. We conclude that no selection towards poly(rA)-containing viral molecules is evident on binding to polyribosomes. These findings suggest that the incoming viral genome of the Moloney strain of murine leukemia virus may serve as a messenger for the synthesis of one or more virus-specific proteins early after infection of mouse fibroblasts.