Serum levels of adiponectin differentiate generalized lipodystrophies from anorexia nervosa.

PURPOSE: The differential diagnosis of lipodystrophy involves other disorders characterized by severe fat loss and may be sometimes challenging. Owing to the rarity of lipodystrophy, it is relevant to search for tools and assays that differentiate it from other diseases that may mimic it. We conducted a study on leptin and high molecular weight (HMW) adiponectin serum concentrations in a series of patients diagnosed with lipodystrophy and compared them with those found in anorexia nervosa, one of the illnesses that may be cause of a missed diagnosis of lipodystrophy. METHODS: Leptin and HMW adiponectin serum concentrations were measured in six patients diagnosed with generalized lipodystrophy (GL), six with progeroid syndromes (PS), 13 with familial partial lipodystrophy type 1 (FPLD1, Kobberling syndrome), 10 with familial partial lipodystrophy type 2 (FPLD2, Dunnigan syndrome), 18 with acquired partial lipodystrophy (APL) and 12 affected by anorexia nervosa (AN). Measurements were compared to those obtained in 12 normal weight healthy subjects. RESULTS: Serum leptin concentrations were reduced to a similar degree in GL, PS and AN, proportionally to the extent of fat loss. Serum concentrations of HMW adiponectin were found extremely low in patients with GL and PS, while comparable to normal weight subjects in patients with AN. CONCLUSION: Serum HMW adiponectin can be regarded as a useful tool to discriminate between generalized lipodystrophy syndromes (including PS) and AN.

Optimisation of X-ray emission from a laser plasma source for the realisation of microbeam in sub-keV region.

In this work, the X-ray emission generated from a plasma produced by focusing Nd-YAG laser beam on the Mylar and Yttrium targets will be characterised. The goal is to reach the best condition that optimises the X-ray conversion efficiency at 500 eV (pre-edge of the Oxigen K-shell), strongly absorbed by carbon-based structures. The characteristics of the microbeam optical system, the software/hardware control and the preliminary measurements of the X-ray fluence will be presented.

Large area extreme-UV lithography of graphene oxide via spatially resolved photoreduction.

The ability to pattern graphene over large areas with nanometer resolution is the current request for nanodevice fabrication at the industrial scale. Existing methods do not match high throughput with nanometer resolution. We propose a high-throughput resistless extreme-UV (EUV) photolithographic approach operating with sub-micrometer resolution on large area (~10 mm(2)) graphene oxide (GO) films via spatially resolved photoreduction. The efficiency of EUV photoreduction is tested with 46.9 nm coherent light produced by a table top capillary discharge plasma source. Irradiated samples are studied by X-ray photoemission spectroscopy (XPS) and micro-Raman Spectroscopy (µRS). XPS data show that 200 mJ/cm(2) EUV dose produces, onto pristine GO, a 6% increase of sp(2) carbon bonds and a 20% decrease of C-O bonds. µRS data demonstrate a photoreduction efficiency 2 orders of magnitude higher than the one reported in the literature for UV-assisted photoreduction. GO patterning is obtained modulating the EUV dose with a Lloyd's interferometer. The lithographic features consist of GO stripes with modulated reduction degree. Such modulation is investigated and demonstrated by µRS on patterns with 2 µm periodicity.

Saponaria officinalis karyology and karyotype by means of image analyzer and atomic force microscopy.

The aim of this work was to offer a contribution to the characterization of taxonomic entity of Saponaria officinalis (2n = 28; an herbaceous perennial species; saporin, a type 1 Ribosome Inactivating Protein, is present in leaves and seeds) by a cytogenetic and karyomorphological approach. We investigated the karyotype's morphometry correlated with Stebbin's symmetric index; the same information has been used for computing the indices resemblance between chromosomes (REC), symmetric indices (SYI), and total form (TF%) which allow the comparison between species and evaluation of karyological evolution. Fluorescence intensities of the stained nuclei were measured by a flow cytometer and, for the first time, values for nuclear DNA content were estimated by comparing nuclei fluorescence intensities of the test population with those of appropriate internal DNA standards. Our study is also aimed to introduce chromosomal volumes, which were determined by atomic force microscopy (AFM), as novel karyomorphological parameter which could allow for chromosome discrimination especially when tiny ones are present.

Damage and ablation of large bandgap dielectrics induced by a 46.9 nm laser beam.

We applied a 0.3 mJ, 1.7 ns, 46.9 nm soft-x-ray argon laser to ablate the surface of large bandgap dielectrics: CaF2 and LiF crystals. We studied the ablation versus the fluence of the soft-x-ray beam, varying the fluence in the range 0.05-3 J/cm2. Ablation thresholds of 0.06 and 0.1 J/cm2 and ablation depths of 14 and 20 nm were found for CaF2 and LiF, respectively. These results define new ablation conditions for these large bandgap dielectrics that can be of interest for the fine processing of these materials.

Interactions between saporin, a ribosome-inactivating protein, and DNA: a study by atomic force microscopy.

Saporins are enzymes belonging to the PNAG class (polynucleotide: adenosine glycosidase), plant enzymes commonly known as ribosome-inactivating proteins (RIP), as a result of their property of irreversibly damaging eukaryotic ribosomes. Direct imaging with tapping-mode atomic force microscopy (AFM) has been used to study pGEM-4Z plasmid DNA binding to the saporin-SO6 (isoform from Saponaria officinalis seeds). Saporin wrapped the plasmidic DNA, and distribution of the enzyme molecules along the DNA chain was markedly variable; plasmid digested with saporin-SO6 appeared fragmented or topologically modified. The supercoiled DNA strands were cleaved, giving rise to a linearized form and to relaxed forms. Electrophoretic analysis of the effect of standard preparations of saporin-SO6 on pGEM-4S confirmed the presence of DNA strand-cleaving activity.

Comparative analysis of isolated cellular organelles by means of soft X-ray contact microscopy with laser-plasma source and transmission electron microscopy.

Soft X-ray contact microscopy (SXCM) is, at present, a useful tool for the examination at submicrometre resolution of biological systems maintained in their natural hydrated conditions. Among current X-ray-generating devices, laser-plasma sources are now easily available and, owing to their pulse nature, offer the opportunity to observe living biological samples before radiation damage occurs, even if the resolution achievable is not as high as with synchrotron-produced X-rays. To assess the potential of laser-plasma source SXCM in the study of cellular organelles, we applied it for the analysis of chloroplasts extracted from spinach leaves and mitochondria isolated from bovine heart and liver. X-ray radiation was generated by a nanosecond laser-plasma source, produced by a single shot excimer XeCl laser focused onto an yttrium target. The images obtained with SXCM were then compared with those produced by transmission electron microscopy observation of the same samples prepared with negative staining, a technique requiring no chemical fixation, in order to facilitate their interpretation and test the applicability of SXCM imaging.

Diffraction of x rays in capillary optics.

Propagation of x rays generated by a small-diameter incoherent source through the capillary waveguide that satisfies the multimode condition is studied with the Fresnel-Kirchhoff diffraction theory. The strong influence of diffraction on the propagation is demonstrated. The diffraction phenomenon is manifested by the appearance of diffraction fringes in both the guide channel and the far-field zone of the capillary output. Experimental data that confirm such behavior of the x-ray radiation is also presented. The results confirm the interference effects recently observed in some experiments on the grazing reflections of x rays in single- and multiple-capillary optics.

[Postoperative peritonitis. The criteria for a reintervention]. / Le peritoniti postoperatorie. I criteri di reintervento.

BACKGROUND: Postoperative peritonitis is a pathologic condition with a sometime nuclear clinical occurrence and therefore with an uncertain timing for reoperation. Aim of this paper is to identify the type and frequency of the digestive and systemic symptoms in relation to the anatomo-pathologic peroperative picture. METHODS: Between 1980 and 1996, 119 patients were reoperated for a postoperative peritonitis (PPO) in the Surgical Department of Modena University. PPO was due to a lesion situated above the mesocolon in 33 patients, from the small bowel in 18, postappendicectomy in 25 and from the colon in 40. The first operation (for benign disease in 66.4%, for malignancy in 33.6%) was performed in emergency in 47 cases (39.5%) and as elective surgery in 72 (60.5%). RESULTS: The global mortality was of 33.6% (40 patients). An attempt is made to identify, the earlier and the most important bioclinical parameters for a correct indication to surgery. Twenty symptoms have been identified that, with different frequency, are strictly related with the onset of a PPO (in average 5 symptoms were positive). A research of these parameters, each 4-6 hours, allow to identify a subclinical PPO. CONCLUSIONS: During the decisional timing, it is important to check these general and digestive symptoms, apparently not serious, in order to avoid the onset appearance of an abdominal tenderness or a multiorgan failure that make the prognosis more severe.

[Myofibroblastoma of the male breast. Apropos of a case]. / Miofibroblastoma della mammella maschile. A proposito di un caso.

A case of male breast myofibroblastoma, personally observed, is described. Preoperative examinations have a limited diagnostic value. Only histological study, with immunohistochemical stain, allows to make a right diagnosis and to distinguish this tumor from other, benign or malignant mesenchymal neoplasms.

Molecular cloning of the human fertilin beta subunit.

Fertilin, a heterodimeric sperm surface protein, first identified in guinea pig, has been shown to play an important role in sperm-egg interactions. We report here the complementary DNA and deduced amino acid sequence of human fertilin beta. The human fertilin beta shares significant sequence homology with mouse, guinea pig and monkey fertilin beta and also exhibits similar structural organization. Of particular interest, the mature guinea pig fertilin beta contains an amino-terminal 90 amino acid disintegrin domain. It has been suggested that the integrin recognition sequence (TDE) of the guinea pig fertilin beta disintegrin domain mediates sperm-egg binding. The amino acid sequence at this position in human fertilin beta differs from the mouse, guinea pig and monkey sequence (mouse-QDE; human-FEE; monkey-FDE).

Long-Duration Soft X-Ray Pulses by XeCl Laser Driven Plasmas and Applications.

We report the characterization of a soft x-ray plasma source generated by a long-pulse XeCl excimer laser system. The output energy is 4 J at a wavelength of 308 nm in a 100-ns pulse. The intensity of radiation on target is estimated to be 4 × 1012 W cm-2. X-ray emission spectra of the plasma have been recorded using a double focusing spatial resolution spectrometer with a spherical mica crystal. From these measurements, the plasma temperature and electron density have been estimated. Various applications of such a plasma source have been investigated. First images of whole intact living cells from our system, imaged using the technique of soft x-ray contact microscopy, utilizing x rays in the "water window" region (280-530 eV), are shown. The suitability of the source for other applications, for example, x-ray lithography and radiation damage studies, to living cells are discussed. Possible improvements to the x-ray source for the various applications are proposed.

An x-ray absorption near edge structure spectroscopy study of metal coordination in Co(II)-substituted Carcinus maenas hemocyanin.

High-resolution x-ray absorption near edge structure spectroscopy was used to characterize the metal sites in three different cobalt-substituted derivatives of Carcinus maenas hemocyanin (Hc), including a mononuclear cobalt, a dinuclear cobalt and a copper-cobalt hybrid derivative. Co(II) model complexes with structures exemplifying octahedral, trigonal bipyramidal, pseudo-tetrahedral, and square planar geometries were also studied. The results provide structural information about the metal binding site(s) in the Co-Hcs that extend earlier results from EPR and optical spectroscopy (Bubacco et al. 1992. Biochemistry. 31: 9294-9303). Experimental spectra were compared to those calculated for atomic clusters of idealized geometry, generated using a multiple scattering approach. The energy of the dipole forbidden 1s-->3d transition and of the absorption edge in the spectra for all cobalt Hc derivatives confirmed the cobaltous oxidation state which rules out the presence of an oxygenated site. Comparisons between data and simulations showed that the mononuclear and dinuclear Co(II) derivatives, as well as the hybrid derivative, contain four-coordinate Co(II) in distorted tetrahedral sites. Although the spectra for Co(II) in dinuclear metal sites more closely resemble the simulated spectrum for a tetrahedral complex than do spectra for the mononuclear derivative, the Co(II) sites in all derivatives are very similar. The Cu K-edge high resolution x-ray absorption near edge structure spectrum of the hybrid Cu-Co-Hc resembles that of deoxy-Hc demonstrating the presence of three-coordinate Cu(I).

Role of glucagon in countering hypoglycemia induced by insulin infusion in dogs.

The aim of the present study was to characterize the role of glucagon in countering the prolonged hypoglycemia resulting from insulin infusion and to determine whether its effect is manifest through glycogenolysis and/or gluconeogenesis. Two groups of 18-h fasted somatostatin-treated dogs were given intraportal insulin at 5 mU.kg-1.min-1. In one group (SimGGN; n = 6), glucagon was infused intraportally so as to mimic the normal response to hypoglycemia. In a second group (BasGGN; n = 6), glucagon was infused at a basal rate. Glucose turnover and gluconeogenesis were assessed by combining tracer and hepatic balance techniques. Exogenous glucose was infused as needed to maintain equivalent hypoglycemia at approximately 45 mg/dl in the two groups. Although glucagon concentrations were significantly different, the levels of other counterregulatory hormones were equivalent in both experimental protocols. Endogenous glucose production (EGP) in SimGGN doubled from 2.4 +/- 0.2 to 5.4 +/- 0.8 mg.kg-1.min-1 by 1 h before dropping to 4.5 +/- 0.2 mg.kg-1.min-1 in the 3rd h of insulin infusion. EGP in BasGGN was initially 2.5 +/- 0.1 mg.kg-1.min-1, unchanged by 1 h, and increased to 3.9 +/- 0.2 mg.kg-1.min-1 by the 3rd h of insulin infusion. In the 1st h of insulin infusion, the rise in gluconeogenesis in both groups was equal and represented only a small part of total EGP. By the 3rd h, gluconeogenesis was the major contributor to total EGP, and gluconeogenic efficiency increased significantly more in SimGGN than BasGGN (261 vs. 140%, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Expression and characterization of the N-terminal half of antistasin, an anticoagulant protein derived from the leech Haementeria officinalis.

Antistasin, a 15-kDa anticoagulant protein isolated from the salivary glands of the Mexican leech Haementeria officinalis, has been shown to be a potent inhibitor of factor Xa in the blood coagulation cascade. Antistasin possesses a twofold internal homology between the N- and C-terminal halves of the molecule, suggesting a gene duplication event in the evolution of the antistasin gene. This structural feature also suggests that either or both halves of the protein may possess biological activity if expressed as separate domains. Because the N-terminal domain contains a factor Xa P1-reactive site, we chose to express this domain in an insect cell baculovirus expression system. Characterization of this recombinant half antistasin molecule reveals that the N-terminal domain inhibits factor Xa in vitro, with a K(i) of 1.7 nM.

Vampire bat salivary plasminogen activator is quiescent in human plasma in the absence of fibrin unlike human tissue plasminogen activator.

The vampire bat salivary plasminogen activator (Bat-PA) is a potent PA that exhibits remarkable selectivity toward fibrin-bound plasminogen (Gardell et al, J Biol Chem 256: 3568, 1989). Herein, we describe the activity of recombinant DNA-derived Bat-PA (rBat-PA) in a human plasma milieu. rBat-PA and recombinant human single-chain tissue plasminogen activator (rt-PA) are similarly efficacious at lysing plasma clots. In stark contrast to rt-PA, the addition of 250 nmol/L rBat-PA to plasma in the absence of a clot failed to deplete plasminogen, alpha 2-antiplasmin and fibrinogen. The lytic activities exhibited by finger-domain minus Bat-PA (F- rBat-PA) and finger and epidermal growth factor-like domains minus Bat-PA (FG- rBat-PA) were less than rBat-PA, especially at low concentrations of PA; nevertheless, these truncated forms also possessed a strict requirement for a fibrin cofactor. The loss of PA activity following the addition of rBat-PA to plasma was slower than that observed when either rt-PA or two-chain rt-PA was added. The efficacy, fibrin selectivity, and decreased susceptibility to inactivation exhibited by rBat-PA in vitro in a human plasma milieu suggests that rBat-PA may be superior to rt-PA for the treatment of thrombotic complications.