The Lectin-Gold Technique: An Overview of Applications to Pathological Problems

Lectins are proteins, mainly of vegetal origin, which recognize glycosidic residues with high specificity; for this property they have been used for many studies of molecular biology. The colloidal gold represents at present the most popular electron dense marker employed in immunocytochemistry, since it offers intrinsic and unique characteristics which are superior to those displayed by the other markers. The cytochemical method which utilizes the gold-labelled lectins takes advantages from both the two systems, in order to optimize the localization of the glycoconjugates. The present paper reviews both the technical aspects of the preparation of the lectin-gold complex and its application to some selected pathological problems. In particular, the papers concerning the eye and ear tissues, the urinary, reproductive, nervous and digestive systems and the blood cells are quoted

Characterization of Mucus Glycoconjugates in Normal Human Conjunctiva by Lectins in Light Microscopy, Transmission and Scanning Electron Microscopy

Maintenance of tear film in normal conditions is dependent on 1) mucus layer integrity and 2) the presence and distribution of conjunctival epithelial cell microvilli. In the present work a new methodology has been developed to gain correlative information about microprojection assessment and mucus composition, from the same specimen, by Light Microscopy (LM), Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM). We have characterized the glycosidic residues secreted by goblet cells in normal human conjunctiva, by means of four lectins (WGA, ConA, PNA and SBA), conjugated with FITC for LM and with colloidal gold for TEM and SEM. The cytochemical reactions were performed on histological sections of paraffin-embedded material and on semithin and ultrathin sections of both Epon embedded material directly processed for TEM and of blocks recovered from SEM and reprocessed for TEM. WGA, ConA, PNA and SBA receptors were found to be constituents of the mucus produced by goblet cells in human conjunctiva. The granules of the so-called second mucus system (SMS) cells were labelled mainly by WGA. A difference in the quality of glycoconjugates between goblet cells and SMS cells has been also demonstrated. Our results provide an improved method to evaluate alterations of tear film that occur in many conjunctival diseases

Relevant and selective activity of Pancratium illyricum L. against Candida albicans clinical isolates: a combined effect on yeast growth and virulence

BACKGROUND: Alkaloids present in plants of the Amaryllidaceae family are secondary metabolites of high biological interest, possessing a wide range of pharmacological activities. In the search for new plant-derived compounds with antimicrobial activities, two alkaloid extracts obtained from bulbs and leaves of Pancratium illyricum L., a plant of the Amarillydaceae family, were tested for their effect on bacterial and yeast growth. METHODS: The broth microdilution susceptibility test was applied to study the effect of plant extracts on the growth of reference bacterial strains and Candida albicans reference and clinical isolates strains. Extracts obtained from the different parts of the plant were tested and compared with the pure components identified in the extracts. Since matrix metalloproteinase enzymes play a role in the dissemination process of Candida albicans, the effect of the bulb extract and pure alkaloids on in vitro collagenase activity was tested. Cell viability test was carried out on human embryo lung fibroblasts (HEL 299). RESULTS: Whilst both extracts did not show any inhibitory activity against neither Gram positive nor Gram negative bacteria, a strong antifungal activity was detected, in particular for the bulb extract. All clinical isolates were susceptible to the growth inhibitory activity of the bulb extract, with endpoint IC50 values ranging from 1.22 to 78 μg/mL. The pure alkaloids lycorine and vittatine, identified as components of the extract, were also assayed for their capacity of inhibiting the yeast growth, and lycorine turned very active, with endpoint IC50 values ranging from 0.89 to 28.5 μg/mL. A potent inhibition of the in vitro collagenase activity was found in the presence of the bulb extract, and this effect was much higher than that exerted by the pure alkaloids. Viability of cell lines tested was not affected by the extract. CONCLUSIONS: Taken together, results suggest that the extract of Pancratium illyricum may act as antifungal agent both directly on the yeast growth and by altering the tissue invasion process

Criminological and Medico-legal Aspects in Homicidal and Suicidal Sharp Force Fatalities

The interpretation of sharp force fatality dynamics may be difficult in some cases, but a contribution to analysis of the phenomenon may be provided by case studies. Therefore, the purpose of our study is focused on identifying, in observed sharp force fatalities, reliable parameters that can differentiate a homicidal and suicidal manner of death, with particular reference to criminological parameters. Data derived from sharp force fatality cases in Padua and Venice from 1997 to 2019, anonymized and collected in Excel, included personal, circumstantial, clinical, and psychopathological\u2013criminological data, as well as crime scene investigation, necroscopic, and toxicological data. Statistical analyses were performed using chi-square and Wilcoxon rank-sum tests. Possible predictors of homicide were analyzed by logistic regression. Six parameters (bloodstains distant from the body, clothing lacerations, hesitation/defense wounds, number of injuries, and potential motives) were significantly different in the two groups (p\ua0<\ua00.05). An independent statistical association between potential motives explaining the crime (p\ua0<\ua00.001; OR 27.533) and homicide on multiple logistic regression analysis was highlighted. The absence of clothing lacerations was inversely related to homicide (p\ua0=\ua00.002, OR 0.092). To the best of our knowledge, this is one of very few Italian studies concerning the differential diagnosis between homicidal and suicidal sharp force fatalities. The dynamics of the event is established in most cases by the integrated evaluation of data from crime scene investigation and the autopsy. Nevertheless, in an atypical scenario, a psychopathological\u2013criminological analysis may provide essential elements, and particular attention should be given to the identification of potential explanatory motives

Correlative Scanning Electron Microscopy in the Study of Human Gastric Mucosa

We studied two aspects of the human gastric mucosa: - the surface morphology of mucous cells, as viewed by scanning electron microscopy (SEM); - the glycosidic components of intracellular mucins, characterized by means of lectins. The latter were conjugated with fluorescein isothiocyanate and with colloidal gold-silver for the visualization of the reaction products in light microscopy (LM) and in SEM (backscattered mode) respectively. The surface morphology of mucous cells appears to be correlated to the secretory state.In gastric ulcers we found a prevalence of non-secreting cells. A decrease in glycosidic receptors for fucose-binding lectin and galactose-(l-3)-N-acetyl-galactosamine-binding lectin was also observed. This suggests the presence of an impaired mucus secretion which may play a role in the pathogenesis of gastric ulcer. Spiral bacteria, supposed to be aetiologically related to peptic ulcer and gastritis, were easily detected by SEM. Intestinal metaplasia defined complete in LM showed surface morphology and glycosidic components different from those of true intestinal mucosa. This implies the necessity of taking into account also these parameters when classifying this lesion. The same applies to polyps. Our data indicate that correlative SEM may contribute further information on the pathogenesis and pathology of gastric diseases

Analysis of short-term blood pressure variability in pheochromocytoma/paraganglioma patients

Data on short-term blood pressure variability (BPV), which is a well-established cardiovascular prognostic tool, in pheochromocytoma and paraganglioma (PPGL) patients is still lack and conflicting. We retrospectively evaluated 23 PPGL patients referred to our unit from 2010 to 2019 to analyze 24 h ambulatory blood pressure monitoring (24-h ABPM)-derived markers of short-term BPV, before and after surgical treatment. PPGL diagnosis was assessed according to guidelines and confirmed by histologic examination. The 24-h ABPM-derived markers of short-term BPV included: circadian pressure rhythm; standard deviation (SD) and weighted SD (wSD) of 24-h, daytime, and night-time systolic and diastolic blood pressure (BP); average real variability (ARV) of 24-h, daytime, and night-time systolic and diastolic BP. 7 males and 16 females of 53 ± 18 years old were evaluated. After surgical resection of PPGL we found a significant decrease in 24-h systolic BP ARV (8.8 ± 1.6 vs. 7.6 ± 1.3 mmHg, p &lt; 0.001), in 24-h diastolic BP ARV (7.5 ± 1.6 vs. 6.9 ± 1.4 mmHg, p = 0.031), and in wSD of 24-h diastolic BP (9.7 ± 2.0 vs 8.8 ± 2.1 mmHg, p = 0.050) comparing to baseline measurements. Moreover, baseline 24-h urinary metanephrines significantly correlated with wSD of both 24-h systolic and diastolic BP. Our study highlights as PPGL patients, after proper treatment, show a significant decrease in some short-term BPV markers, which might represent a further cardiovascular risk factor

Metabolomic analysis and bioactivities of Arbutus unedo leaves harvested across the seasons in different natural habitats of Sardinia (Italy)

Background Arbutus unedo L. is a wild tree of Mediterranean regions used as food and in traditional medicine and important for afforestation programs. There is no detailed information available on the variation of A. unedo leaves metabolome across the seasons. The leaves were analyzed by Proton nuclear magnetic resonance ((1) H NMR)-based metabolomics, comparing samples harvested across the seasons and in ten different natural habitats of Sardinia (Italy).Results Multivariate analysis showed the impact of seasonal variation on the metabolome: glucose and quinic acid increased in summer, while in spring sucrose was accumulated. beta-Arbutin, the main known active principle of A. unedo, generally reached the highest concentration in autumn. In winter, O-beta-methylglucose, gamma-aminobutyric acid (GABA), flavonols (quercetin-3-O-alpha-rhamnoside, myricetin-3-O-alpha-rhamnoside, kaempferol-3-O-alpha-rhamnoside), catechin, and gallocatechin increased. Characteristic metabolomic features were found also for samples collected in different locations. For instance, trees growing at the highest altitude and exposed to lower temperatures produced less flavonols and catechins. The only sample collected on trees growing on limestones, dolomites, and dolomitic limestones type of soil showed generally the highest content of arbutin. The highest phenolics content was found during spring, while samples collected on flowering branches in winter were the ones with the highest flavonoid content. The antioxidant activity was also variated, ranging from 1.3 to 10.1 mg of Trolox equivalents (TE)/mL of extract, and it was positively correlated to both total phenolics and flavonoid content. Winter samples showed the lowest antibacterial activity, while summer and autumn ones exhibited the highest activity (IC50 values ranging from 17.3 to 42.3 mu g/mL against Staphylococcal species).Conclusion This work provides H-1 -NMR fingerprinting of A. unedo leaves, elucidating the main metabolites and their variations during seasons. On the basis of arbutin content, autumn could be considered the balsamic period of this taxon. Samples collected in this season were also the most active ones as antibacterial. Moreover, an interesting metabolomic profile enriched in catechins and flavonols was observed in leaves collected in winter on flowering branches which were endowed with high antioxidant potential

Mutation within TARDBP leads to Frontotemporal Dementia without motor neuron disease.

2009 AUG 4. [EPUB AHEAD OF PRINT

Importance of Tests for the Complete Lorentz Structure of the t --> W+ b vertex at Hadron Colliders

The most general Lorentz-invariant decay-density-matrix for $t\to W^{+}b\to (l^{+}\nu)b$, or for $t\to W^{+}b\to (j_{\bar d}j_u)b$, is expressed in terms of eight helicity parameters. The parameters are physically defined in terms of partial-width-intensities for polarized-final-states in $t\to W^{+}b$ decay. The parameters are the partial width, the $b$ quark's chirality parameter $\xi$, the $W^+$ polarimetry parameter $\sigma$, a "pre-SSB" test parameter $\zeta$, and four $W_{L}$-$W_{T}$ interference parameters $\eta$, $\eta^{'}$, $\omega$, $\omega^{'}$ which test for $\tilde T_{FS}$ violation. They can be used to test for non-CKM-type CP violation, anomalous $\Gamma_{L,T}$'s, top weak magnetism, weak electricity, and second-class currents. By stage-two spin-correlation techniques, percent level statistical uncertainites are typical for measurements at the Tevatron, and several mill level uncertainites are typical at the LHC.Comment: Minor clarifications. Expression for r_{+-} corrected. 19 pages LaTex + Tables + 1 Figur

High Resolution Hybrid Pixel Sensors for the e+e- TESLA Linear Collider Vertex Tracker

In order to fully exploit the physics potential of a future high energy e+e- linear collider, a Vertex Tracker, providing high resolution track reconstruction, is required. Hybrid Silicon pixel sensors are an attractive option, for the sensor technology, due to their read-out speed and radiation hardness, favoured in the high rate environment of the TESLA e+e- linear collider design but have been so far limited by the achievable single point space resolution. In this paper, a conceptual design of the TESLA Vertex Tracker, based on a novel layout of hybrid pixel sensors with interleaved cells to improve their spatial resolution, is presented.Comment: 12 pages, 5 figures, to appear in the Proceedings of the Vertex99 Workshop, Texel (The Netherlands), June 199