Application of spectrometric analysis to the identification of pollution sources causing cultural heritage damage.

Black crusts are recognized to have been, up to now, one of the major deterioration forms affecting the built heritage in urban areas. Their formation is demonstrated to occur mainly on carbonate building materials, whose interaction with an SO2-loaded atmosphere leads to the transformation of calcium carbonate (calcite) into calcium sulfate dihydrate (gypsum) which, together with embedded carbonaceous particles, consequently forms the black crusts on the stone surface. An analytical study was carried out on black crust samples collected from limestone monumental buildings and churches belonging to the European built Heritage, i.e., the Corner Palace in Venice (Italy), the Cathedral of St. Rombouts in Mechelen (Belgium), and the Church of St. Eustache in Paris (France). For a complete characterization of the black crusts, an approach integrating different and complementary techniques was used, including laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), Fourier transform infrared spectroscopy, optical and scanning electron microscopy. In particular, the application of LA-ICP-MS permitted to obtain a complete geochemical characterization in terms of trace elements of the black crusts from the inner parts to the external layers contributing to the identification of the major combustion sources responsible for the deterioration over time of the monuments under study. In addition, the obtained results revealed a relation between the height of sampling and the concentration of heavy metals and proved that the crust composition can be a marker to evaluate the variation of the fuels used over time.

A review of bacteriocinogenic lactic acid bacteria used as bioprotective cultures in fresh meat produced in Argentina.

Several lactic acid bacteria (LAB) associated with meat products are important natural bacteriocin producers. Bacteriocins are proteinaceous antagonistic substances that are important in the control of spoilage and pathogenic microorganisms. The use of LAB as bioprotective cultures to extend the shelf life of fresh meat can improve microbial stability and safety in commercial meat preservation. Lactobacillus curvatus CRL705 used as a protective culture in fresh beef is effective in inhibiting Listeria innocua and Brochothrix thermosphacta as well as the indigenous contaminant LAB, retaining its inhibitory effect at low temperatures and having a negligible effect on meat pH. In addition to the hurdle represented by low temperature and vacuum-packaging, the use of live cells of Lb. curvatus CRL705 seems more feasible from an economic point of view - and without legal restrictions - compared to the addition of purified bacteriocins. A description of meat-borne bacteriocins and their application in meat to extend shelf life is discussed.

Reduction of Escherichia coli population following treatment with bacteriocins from lactic acid bacteria and chelators.

The inhibitory activity of lactocin 705/AL705 (2133 arbitrary units per ml (AU ml(-1))), two bacteriocins produced by Lactobacillus curvatus CRL705 and nisin (1066AU ml(-1)) produced by Lactococcus lactis CRL1109 in combination with chelating agents against Escherichia coli strains in TSB medium at 21 and 6 degrees C was investigated. Treatment with EDTA (500 and 1000 mm) and Na lactate (800 mm) alone produced a variable effect depending on the strain, Na lactate being inhibitory against E. coli NCTC12900 at both assayed temperatures while EDTA (1000 mm) led to its inactivation only at 6 degrees C. Direct and deferred strategies using EDTA and Na lactate showed that the direct addition of bacteriocins and chelators was not as effective as compared to deferred treatments. When the deferred treatment effectiveness was evaluated at 6 degrees C, the use of EDTA (500 and 1000 mm) and Na lactate (800 mm) in combination with lactocin 705/AL705 demonstrated to be the most inhibitory strategy against both E. coli strains. Nevertheless, treatments with chelators and bacteriocins was highly dependent upon strain sensitivity. Permeabilization of the outer membrane of E. coli strains with EDTA and Na lactate combined with lactocin 705/AL705 showed to be valuable in controlling this foodborne bacteria at low temperatures.

[Mininvasive treatment of strangulated paraesophageal hiatal hernia in emergency. Case report]. / Trattamento chirurgico minivasivo d'urgenza di ernia gastrica iatale paraesofagea strozzata. Caso clinico.

The Authors report a case of strangulated paraesophageal hiatal hernia occurred in a elderly woman and treated with laparoscopic approach. After review of the literature regard on this uncommon pathology that present about 5% of the hiatal hernias, they emphasize that the laparoscopic approach is appropriated even in emergency and comprises complete reduction of the stomach in abdomen, control of suitable position of the distal esophagus and cardias and making of effective hiatus-plasty.

[A fecal fistula in patient who underwent an abdomino-perineal resection with a continent magnetic prosthesis: observations on an obsolete technique]. / Fistola stercoracea in paziente sottoposta ad intervento di Miles e portatrice di protesi magnetica: considerazioni su una tecnica obsoleta.

The Authors consider a case of a patient who underwent an abdominal-perineal resection and presented a fecal fistula as a late complication of a magnetic prosthesis implant. After a revision of the literature, the Authors evaluate the reasons for abandoning this surgical technique of continence many years ago, underlying not only the complications observed during the experimentation but also the lack of those benefits for which this technique was proposed.

[Pelvic abscesses. Treatment modalities and personal experience]. / Gli ascessi pelvici. Modalità di trattamento ed esperienza personale.

The pelvic abscesses as complication of surgical operations or various pathologies is a delicate clinical situation because of the possible and unexpected evolution into a settic shock with a high risk of death. The authors report their personal experience of 16 cases undergone surgical treatment. They underline the importance of an early diagnosis, of a control of the patient general conditions with a specific antibiotic therapy, of the possibility to treat this pathology in a percutaneous way with the help of radiologic techniques or eventually by surgery.

Identification and discrimination of Electrogena species by numerical methods (Ephemeroptera: Heptageniidae).

Nine species of Electrogena from thirty-one populations were investigated on the basis of five meristic and five ratio characters of the larvae. The attribution of populations to species was checked by a cluster analysis based on the generalized distances between pairs. Groups representing species were then subjected to a multiple discriminant analysis; discriminant functions and loadings were calculated. The correct attribution of individuals to species by the discriminant functions ranged from 97% to 100%. The analysis of discriminant loadings shows which characters contribute more to the discrimination of species. Although many species could be identified with some confidence by a sequential monothetic key based on meristic and qualitative characters, the discriminant analysis improves the effectiveness of identification of all species.

Regulation of cytochrome P450scc synthesis and activity in the ovine corpus luteum.

The rate-limiting step in luteal biosynthesis of progesterone consists of cleavage of the side chain of cholesterol by mitochondrial cytochrome P450 side-chain cleavage enzyme (P450scc) to form pregnenolone. Luteal mRNA encoding P450scc, quantitated on selected days of the 16-day ovine estrous cycle, was similar on days 3 and 6, increased by 2-fold on day 9 (P < 0.05) and remained elevated on day 15. Levels of P450scc mRNA on day 15 of pregnancy were not different from those found on any day of the cycle (P < 0.05). To determine whether levels of mRNA encoding P450scc are hormonally regulated, ewes on day 10 of the estrous cycle were injected with hCG or prostaglandin F2 alpha (PGF2 alpha). P450scc mRNA was not increased for up to 36 h after injection of hCG, nor decreased within 8 h after injection of PGF2 alpha (P < 0.05). An assay for P450scc activity was developed which utilized ovine small and large luteal cells in the presence of 22R-hydroxycholesterol and ovine high density lipoprotein. Enzyme activity was quantitated by measurement of progesterone production. In small luteal cells activation of the protein kinase A (PKA) second-messenger system by treatment with LH resulted in 910% increase in progesterone production without altering activity of P450scc. Activation of the protein kinase C (PKC) second-messenger system with phorbol 12-myristate 13-acetate caused a 51% reduction in progesterone secretion from large luteal cells but did not alter activity of P450scc. These findings suggest that in mature luteal tissue steady state levels of mRNA encoding P450scc, and enzyme activity are independent of acute regulation by activation of PKA or PKC second-messenger systems.

A specific binding protein from Tenebrio molitor for the insecticidal toxin of Bacillus thuringiensis subsp. tenebrionis.

Biopesticides based on the bacterium Bacillus thuringiensis have attracted wide attention as safe alternatives to chemical insecticides. In this paper, we report, for the first time, the identification of a single binding protein from a coleopteran insect, Tenebrio molitor, that is specific for the cryIII toxin of B. thuringiensis. The protein appeared as a single band of 144 kDa on radioligand and immunoblots of total proteins extracted from brush border membrane vesicles of the midgut of T. molitor. Radiolabelled cryIIIA toxin bound to the protein with a Kd value of 17.5 nM and could be specifically blocked by unlabelled toxin but not by toxins from other subspecies of B. thuringiensis. This study lays the groundwork to clone the cryIIIA toxin binding protein and to determine the molecular mechanism(s) of toxin action.

Luteal function: the estrous cycle and early pregnancy.

A number of morphological and biochemical changes occur as the cells of the recently ovulated follicle luteinize and develop into a functional CL. There are two distinct steroidogenic luteal cell types that appear to differentiate from thecal and granulosal cells in the follicle. The control of progesterone secretion is quite different in the two cell types. Prostaglandin F2 alpha (PGF2 alpha) is the primary luteolytic hormone in most mammals. PGF2 alpha appears to exert its antisteroidogenic actions via activation of the protein kinase C system, while its cytotoxic effects appear to be mediated via a dramatic increase in intracellular levels of free calcium. The mechanisms involved in maternal recognition of pregnancy are very diverse between species and may involve direct luteotropic stimulation of the CL, reduced uterine secretion of PGF2 alpha, and/or inhibition of actions of PGF2 alpha at the level of the CL.

Steroidogenic enzyme activity after acute activation of protein kinase (PK) A and PKC in ovine small and large luteal cells.

Intracellular effector systems which utilize PKA and PKC can be pharmacologically activated by forskolin and phorbol 12-myristate 13-acetate (PMA) and appear to be important for regulation of steroidogenesis by cells of the corpus luteum. In this study the effect of pharmacologic activation of PKA (forskolin) or PKC (PMA) on the activity of adenylate cyclase, cholesterol esterase, P450 cholesterol side chain cleavage (P450scc) and 3 beta-hydroxysteroid dehydrogenase/delta 5, delta 4 isomerase (3 beta HSD) was determined. Basal adenylate cyclase activity (as measured by intracellular and secreted cAMP) was extremely low in both large and small luteal cells. Forskolin stimulated adenylate cyclase activity in both large and small luteal cells but progesterone production was increased only in small cells. PMA inhibited progesterone production by large and forskolin-stimulated small cells without altering adenylate cyclase activity. Basal cholesterol esterase activity was greater in small than in large cells and was stimulated by forskolin only in small cells. PMA did not significantly alter cholesterol esterase activity in either cell type. Activity of P450scc or 3 beta HSD was measured by conversion of hydroxylated cholesterol derivatives (P450scc) or pregnenolone (3 beta HSD) to progesterone. Although basal progesterone production was 47 times greater in large than small cells, there was only 5.1 (P450scc) and 6.4 (3 beta HSD) times greater enzyme activity in large than in small luteal cells. Activation of PKA and/or PKC did not alter the activity of P450scc or 3 beta HSD in either cell type.(ABSTRACT TRUNCATED AT 250 WORDS)

Regulation of messenger ribonucleic acid encoding 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase in the ovine corpus luteum.

Three experiments were conducted to determine how steady-state levels of mRNA encoding 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase (3 beta-HSD) in the ovine corpus luteum vary 1) between the two steroidogenic luteal cell types, 2) during the estrous cycle, and 3) during prostaglandin F2 alpha (PGF2 alpha)-induced luteolysis. In the first experiment, RNA (10 micrograms) was isolated from purified preparations (n = 4) of large or small steroidogenic luteal cells. Large luteal cells contained 42% more (p < 0.05) message for 3 beta-HSD per microgram RNA than did small luteal cells, while the amount of mRNA for tubulin did not differ between the two types of luteal cells. To determine whether luteal levels of mRNA for 3 beta-HSD differ during the estrous cycle, corpora lutea were collected from cycling ewes (n = 3/day) on Days 3, 6, 9, 12, and 15 postestrus. Levels of mRNA for 3 beta-HSD were similar on Days 3, 6, 9, and 12 but were lower (p < 0.05) on Day 15 postestrus, while levels of mRNA for tubulin were unchanged. In the final experiment, ewes were treated on Day 10 postestrus with two injections of PGF2 alpha (5 mg each) or saline (control) at a 4-h interval. Corpora lutea were collected from ewes (n = 4/treatment) 1 h or 8 h after the second injection of PGF2 alpha or 8 h after the second saline injection.(ABSTRACT TRUNCATED AT 250 WORDS)

Proteins secreted from the early ovine conceptus block the action of prostaglandin F2 alpha on large luteal cells.

In this study we evaluated whether the early conceptus secretes a factor that blocks the action of prostaglandin (PG) F2 alpha on cultured ovine large luteal cells. PGF2 alpha inhibited progesterone production by lipoprotein-stimulated large luteal cells and this anti-steroidogenic action was blocked in a dose-dependent manner by conceptus proteins secreted from Day 15 embryos. Purified ovine trophoblast protein-1 (oTP-1) did not exhibit the anti-PGF2 alpha activity, but secreted conceptus proteins devoid of oTP-1 did prevent the anti-steroidogenic effects of PGF2 alpha. This activity does not appear to be a nonspecific effect of protein since neither serum albumin nor thyroglobulin, gamma globulin, insulin, LH, secreted ovine endometrial proteins, or heat-inactivated secreted conceptus proteins had this action. After molecular-sizing chromatography we found a high- and a low-molecular weight fraction with luteal protective activity. Neither of the secreted conceptus protein fractions blocked the binding of 3H-PGF2 alpha to large luteal cells. However, conceptus proteins did block the anti-steroidogenic action of phorbol ester and calcium ionophore on large luteal cells, suggesting that secreted conceptus proteins act after activation of the free calcium/protein kinase C intracellular effector pathways. Thus, the early ovine conceptus secretes a luteal protective protein(s) that may be important for maintaining the corpus luteum during early pregnancy; however, the physiologic significance of this luteal protective protein(s) cannot be stated without further investigation.