Evaluation of the Implementation of Good Handling Practices in Food and Beverage Areas of Hotels.

Because of the major international-level events that have recently been held in Brazil, concerns about the sensory and hygienic-sanitary conditions of food have increased. The objective of this study was to evaluate the implementation of good handling practices in food and beverage areas of hotels, with and without outsourced professional intervention. We evaluated 19 food and beverage areas in hotels in Porto Alegre, Rio do Sul, Brazil, using a checklist that was developed by a municipal surveillance team based on existing laws for good handling practices. The evaluation was done by a skilled professional in the food safety area on two occasions, at the beginning of the study (January to May 2013) and at the end (July to November 2014), and the establishments were classified as good, regular, or poor. After the baseline evaluation, an action plan listing the noncompliance found at each location was given to those responsible for the establishments, and a period of 1 year 6 months was stipulated for improvements to be made. In the repeat evaluation, those responsible for the establishments were asked whether they had hired an outsourced professional to assist them in the improvements. The hotels showed improvement during the repeat evaluation, but a significant increase in the percentage of overall adequacy was seen only in the food and beverages areas of the 12 hotels that used the intervention of an outsourced professional. The better percentage of adequacy in establishments with outsourced professional intervention underlines the importance of an external and impartial view of routine activities in the implementation of good handling practices.

The human Rgr oncogene is overexpressed in T-cell malignancies and induces transformation by acting as a GEF for Ras and Ral.

The Ras superfamily of GTPases is involved in the modification of many cellular processes including cellular motility, proliferation and differentiation. Our laboratory has previously identified the RalGDS-related (Rgr) oncogene in a DMBA (7,12-dimethylbenz[α]anthracene)-induced rabbit squamous cell carcinoma and its human orthologue, hRgr. In this study, we analyzed the expression levels of the human hRgr transcript in a panel of human hematopoietic malignancies and found that a truncated form (diseased-truncated (Dtr-hrgr)) was significantly overexpressed in many T-cell-derived neoplasms. Although the Rgr proto-oncogene belongs to the RalGDS family of guanine nucleotide exchange factors (GEFs), we show that upon the introduction of hRgr into fibroblast cell lines, it is able to elicit the activation of both Ral and Ras GTPases. Moreover, in vitro guanine nucleotide exchange assays confirm that hRgr promotes Ral and Ras activation through GDP dissociation, which is a critical characteristic of GEF proteins. hRgr has guanine nucleotide exchange activity for both small GTPases and this activity was reduced when a point mutation within the catalytic domain (CDC25) of the protein, (cd) Dtr-hRgr, was utilized. These observations prompted the analysis of the biological effects of hRgr and (cd) hRgr expression in cultured cells. Here, we show that hRgr increases proliferation in low serum, increases invasion, reduces anchorage dependence and promotes the progression into the S phase of the cell cycle; properties that are abolished or severely reduced in the presence of the catalytic dead mutant. We conclude that the ability of hRgr to activate both Ral and Ras is responsible for its transformation-inducing phenotype and it could be an important contributor in the development of some T-cell malignancies.

Chromium geochemistry and bioaccumulation in sediments from the lower Hackensack River, New Jersey.

Total and hexavalent chromium [Cr(VI)] were measured in sediment and sediment porewater in the lower Hackensack River (NJ) to assess the relationship between sediment geochemistry and chromium speciation, which in turn controls the mobility, bioavailability, and toxicity of chromium. Between 2003 and 2005, >100 surface (0 to 15 cm) sediment samples were tested for total chromium and Cr(VI), acid-volatile sulfides (AVS), ferrous iron (Fe(II)), divalent manganese (Mn(II)), ammonia, and organic carbon. Sediment porewater samples were collected by centrifugation or using in situ samplers colocated with the collection of sediments. In whole sediments, total chromium and Cr(VI) concentrations ranged from 5 to 9190 mg/kg dry weight (dw) and from <0.47 to 31 mg/kg dw, respectively. Sediment porewater concentrations ranged from <10 to 83 microg/l for total chromium; Cr(VI) was not detected in sediment porewater (n = 78). Concentrations of AVS (ranging between <10.6 to 4178 mg/kg) and other geochemistry measurements indicated anoxic, reducing conditions in the majority of sediment samples. In polychaetes (Nereis virens) and clams (Macoma nasuta) exposed in the laboratory for 28 days to sediments contained between 135 and 1780 mg/kg dw total chromium, concentrations in whole tissues after 24-hour depuration ranged between 1.2 and 14.8 mg/kg wet weight (ww; median 1.6 mg/kg ww) total chromium. In whole tissues of indigenous polychaetes collected from the sediment, tissue concentrations of total chromium ranged between 1.0 and 37.5 mg/kg ww (median = 2.1 mg/kg ww). Chromium concentrations in whole tissues of animals exposed in the field or in the laboratory showed no relationship with total chromium or Cr(VI) concentrations in the sediment. There were no statistical differences among animals exposed to sediments from site and reference locations. The results of this study are consistent with sediment studies conducted elsewhere indicating low chromium bioavailability in sediment under reducing conditions. This study also highlights the importance of sediment geochemistry and in situ porewater measurements to understand the ecological significance of chromium in sediment and the potential for human health and ecological exposures.

The relationship between Taxol and (+)-discodermolide: synthetic analogs and modeling studies.

BACKGROUND: During the past decade, Taxol has assumed an important role in cancer chemotherapy. The search for novel compounds with a mechanism of action similar to that of Taxol, but with greater efficacy particularly in Taxol-resistant cells, has led to the isolation of new natural products. One such compound, (+)-discodermolide, although structurally distinct from Taxol, has a similar ability to stabilize microtubules. In addition, (+)-discodermolide is active in Taxol-resistant cell lines that overexpress P-glycoprotein, the multidrug-resistant transporter. Interestingly, (+)-discodermolide demonstrates a profound enhancement of the initiation process of microtubule polymerization compared to Taxol. RESULTS: The synthesis of (+)-discodermolide analogs exploiting our highly efficient, triply convergent approach has permitted structure-activity relationship (SAR) studies. Small changes to the (+)-discodermolide structure resulted in a dramatic decrease in the ability of all four discodermolide analogs to initiate tubulin polymerization. Two of the analogs also demonstrated a decrease in total tubulin polymerization, while a change in the olefin geometry at the C8 position produced a significant decrease in cytotoxic activity. CONCLUSIONS: The availability of (+)-discodermolide and the analogs, and the resultant SAR analysis, have permitted an exploration of the similarities and differences between (+)-discodermolide and Taxol. Docking of the X-ray/solution structure of (+)-discodermolide into the Taxol binding site of beta-tubulin revealed two possible binding modes (models I and II). The preferred pharmacophore model (I), in which the C19 side chain of (+)-discodermolide matches with the C2 benzoyl group of Taxol and the delta-lactone ring of (+)-discodermolide overlays with the C13 side chain of Taxol, concurred with the results of the SAR analysis.

Resistance to Taxol in lung cancer cells associated with increased microtubule dynamics.

Microtubule dynamics are crucial for mitotic spindle assembly and chromosome movement. Suppression of dynamics by Taxol appears responsible for the drug's potent ability to inhibit mitosis and cell proliferation. Although Taxol is an important chemotherapeutic agent, development of resistance limits its efficacy. To examine the role of microtubule dynamics in Taxol resistance, we measured the dynamic instability of individual rhodamine-labeled microtubules in Taxol-sensitive and -resistant living human cancer cells. Taxol-resistant A549-T12 and -T24 cell lines were selected from a human lung carcinoma cell line, A549. They are, respectively, 9- and 17-fold resistant to Taxol and require low concentrations of Taxol for proliferation. We found that microtubule dynamic instability was significantly increased in the Taxol-resistant cells. For example, with A549-T12 cells in the absence of added Taxol, microtubule dynamicity increased 57% as compared with A549 cells. The length and rate of shortening excursions increased 75 and 59%, respectively. These parameters were further increased in A549-T24 cells, with overall dynamicity increasing by 167% compared with parental cells. Thus, the decreased Taxol-sensitivity of these cells can be explained by their increased microtubule dynamics. When grown without Taxol, A549-T12 cells were blocked at the metaphase/anaphase transition and displayed abnormal mitotic spindles with uncongressed chromosomes. In the presence of 2-12 nM Taxol, the cells grew normally, suggesting that mitotic block resulted from excessive microtubule dynamics. These results indicate that microtubule dynamics play an important role in Taxol resistance, and that both excessively rapid dynamics and suppressed dynamics impair mitotic spindle function and inhibit proliferation.

Combined effects of pentachlorophenol and salinity stress on chemiluminescence activity in two species of abalone.

The effect of pentachlorophenol (PCP) combined with salinity stress on hemocyte microbicidal activity was examined in two species of abalone. Microbicidal phagocytic function was determined in red (Haliotis rufescens) and black (Haliotis cracherodii) abalone after in vivo exposure to 25, 35 and 45 per thousand seawater salinity plus 1.2 mg/l PCP using luminol-dependent chemiluminescence (CL). Red and black abalone exposures of 3.5 and 6.5 h, respectively, were based on species-specific metabolic endpoints (MEPs) derived from previous nuclear magnetic resonance spectroscopy (NMR) data. Endpoints examined include total CL (CL(total)), peak CL (CL(max)), and the time to reach peak CL (T(max)). Overall, black abalone CL was significantly greater than red abalone CL particularly at ambient and high salinities. High salinity alone had a dramatic effect on red abalone whereas black abalone demonstrated few salinity effects. While the addition of PCP stimulated CL(max) and CL(total) among red abalone at ambient and high salinities, PCP exposure inhibited CL(max) at each salinity and inhibited CL(total) at ambient salinity among black abalone. Black abalone generally did not demonstrate effects of PCP within the 3.5 h exposure period except at high salinity plus PCP, which caused a reduction of CL(total). T(max) was greatly increased after PCP exposure at each salinity among red abalone but did not effect T(max) at any salinity tested among black abalone. No lysozyme activity was detected among red or black abalone after exposure to any of four different target particles tested either in the presence or absence of PCP. Overall, PCP in combination with salinity stress causes a modulation in the production of reactive oxygen species and this modulation varies between abalone species. Agents that decrease CL activity in hemocytes may reduce the antimicrobial potential of these cells thereby increasing susceptibility to infectious disease.

Taxol and discodermolide represent a synergistic drug combination in human carcinoma cell lines.

Recently, three natural products have been identified, the epothilones, eleutherobin, and discodermolide, whose mechanism of action is similar to that of Taxol in that they stabilize microtubules and block cells in the mitotic phase of the cell cycle. In this report, we have compared and contrasted the effects of these new agents in Taxol-sensitive and -resistant cell lines. We also have taken advantage of a human lung carcinoma cell line, A549-T12, that was isolated as a Taxol-resistant cell line and found to require low concentrations of Taxol (2-6 nM) for normal cell division. This study then examined the ability of these new compounds to substitute for Taxol in sustaining the growth of A549-T12 cells. Immunofluorescence and flow cytometry have both indicated that the epothilones and eleutherobin, but not discodermolide, can substitute for Taxol in this Taxol-dependent cell line. In A549-T12 cells, the presence of Taxol significantly amplified the cytotoxicity of discodermolide, and this phenomenon was not observed in combinations of Taxol with either the epothilones or eleutherobin. Median effect analysis using the combination index method revealed a schedule-independent synergistic interaction between Taxol and discodermolide in four human carcinoma cell lines, an effect that was not observed between Taxol and epothilone B. Flow cytometry revealed that concurrent exposure of A549 cells to Taxol and discodermolide at doses that do not induce mitotic arrest caused an increase in the hypodiploid population, thereby indicating that a possible mechanism for the observed synergy is the potentiation of apoptosis. Our results suggest that Taxol and discodermolide may constitute a promising chemotherapeutic combination.

Regulation of electrolyte transport by nitric oxide in the mouse cecum.

The effect and role of nitric oxide (NO) in the regulation of ion transport in the mouse cecum were investigated. L-arginine, used to increase NO production, increased short-circuit current (Isc), a measure of ion transport, in a concentration-dependent manner with a maximal increase of 193.8+/-65.5 microA/cm2. This increase was not changed in Cl-- or HCO3--free buffers, but was significantly decreased in Na+-free buffer. Using immunohistochemistry, the constitutive form of nitric oxide synthase was found not to be different in the inflamed cecum. The inducible form of the enzyme, however, which was absent in the cecum of normal mice, was present in high levels in the cecum of the colitic mouse. These results suggest that NO causes an increase in Na+ absorption. The increased levels of inducible NO synthase in the inflamed cecum suggest a role for NO in the pathophysiology of inflammatory bowel disease.

Morphological factors influencing transepithelial conductance in a rabbit model of ileitis.

BACKGROUND & AIMS: Infection of rabbits with coccidia (Eimeria magna) causes chronic ileal inflammation and diarrhea. Inflamed ileum also shows decreased transmural conductance. The aim of this study was to characterize morphological factors known to affect paracellular permeability that may alter transmural conductance in inflamed ileum. METHODS: Ileal mucosa was mounted in Ussing chambers for study of [3H]mannitol and [3H]inulin fluxes. Light and electron microscopy were used for morphometric studies. Alterations in the zonula occludens of epithelial cells were evaluated in freeze-fracture replicas. RESULTS: Inflamed ileum showed diminished paracellular fluxes. Inoculated rabbits showed marked lymphoplasmocytic infiltration and villus blunting in ileum. Villus linear junctional density was unaffected. However, total villus apical surface area per square centimeter of tissue was reduced in inflamed ileum, causing a diminished total villus linear junctional pathway per square centimeter of apical surface. Villus zonula occludens strand number was reduced in inflamed ileum, whereas the frequency of both villus and crypt lateral surface extrajunctional strands increased. CONCLUSIONS: Chronic inflammation exerts a profound effect on ileal paracellular permeability. Morphological data suggest that this effect may be caused in part by alterations in inflamed ileal mucosal structure and tight junctional organization and density, particularly on villi.

Identification of protein kinase C-alpha, epsilon and zeta in rabbit ileal enterocytes.

Activation of protein kinase C (PKC) has been shown to regulate electrolyte transport in rabbit small intestine. We investigated the types of PKC isoforms in rabbit ileal villus and crypt cells and the potential for phorbol 12-myristate 13-acetate (PMA) to induce translocation of the PKC from the cytosol to the plasma membrane. Our results indicate that there are at least three PKC isoforms, alpha, epsilon and zeta, are present in both villus and crypt enterocytes. Acute treatment with PMA resulted in translocation of the PKC-alpha from the cytosol to the membrane fraction in both cell types. Prolonged exposure of the villus cells to PMA resulted in a significant progressive decrement in PKC-alpha, suggesting down regulation. Since PMA treatment results in translocation, this isoform may be involved in the regulation of electrolyte transport in the rabbit ileum.

[Prevention of vena cava thrombosis during catheterization. Comparison of calciheparin and defibrotide]. / Profilassi della trombosi venosa cavale in corso di cateterizzazione. Confronto tra calcieparina e defibrotide.

Total intraparenteral nutrition therapy using an endocaval catheter is subject to complications due to the catheter itself and to the onset of phlebothrombosis of the venous district involved. Two groups of patients suffering from abdominal surgical pathologies requiring prolonged postoperative parenteral treatment were subjected to antithrombotic prophylaxis. In the first group of 20 patients, calciheparin u.s. was used and in the second defibrotide in the doses recommended in the literature. Serial lab screening of certain clotting parameters was carried of the brachio-subclavio-caval district. In the calciheparin group, the trend of certain examinations shows, as usual, a change in parameters in agreement with the drug's anticlotting action; in the group treated with defibrotide, the haemostatic balance is respected: the angiographies never showed intimal lesion or phlebothrombosis in either group. The usefulness of correct antithrombotic prophylaxis in these patients is confirmed and it is pointed out that defibrotide is more flexible and handy and that it can be used in patients in whom calciheparin is potentially risk.

Augmentation mentoplasty: a critical analysis.

An evaluation of the senior author's mentoplasty technique by use of inorganic silicone and acrylic implants is presented, encompassing 16 years of experience. The evaluation was chiefly concerned with the silicone implant and long-term followup observations of its survival and some of its extrusions and complications.

[Total parenteral nutrition and dialysis. Complementary importance in the therapy of acute renal insufficiency in aortic surgery]. / Nutrizione parenterale total (N.P.T.) e dialisi. Momenti complementari della terapia della insufficienza renale acuta (I.R.A.) nella chirurgia dell'aorta.

The acute renal insufficiency (A.R.I.) consequent on surgical pathology of the aorta is in the Milan School, one of the most important postoperative complications and requires the use of total parenteral feeding (T.P.F.). Parenteral infusion of AAe and hypertonic glucose in patients with A.R.I. has given positive results, not only insofar as it improves the general nutritional state, but also because it facilitates recovery of renal function and improves survival. Reutilisation of endogenous nitrogen gives a synthesis of structural proteins to the benefit of metabolic homoeostasis and the patient's clinical condition. On the basis of recent nephrology studies on uraemic toxicity, the therapeutic problem of A.R.I. in surgical patients has been examined: early peritoneal dialysis associated with T.P.F. and the combination, in the postoperative stage, of parenteral feeding and periodic peritoneal dialysis. The average duration of this treatment has been personally found to be about 8-18 days. The example is given of a clinical case of A.R.I. in a patient operated by aneurysmectomy for rupture of an aneurysm of the abdominal aorta.

[Comparative evaluation of local 02 consumption in lumbar sympathetic infiltration as compared with acupuncture]. / Valutazione comparativa del consumo di O2 distrettuale con l'infiltrazione del simpatico lombare e la riflessoterapia (agopuntura)

The effectiveness of acupuncture in the treatment of lower limb obliterating arteritis as an alternative to pharmacological sympatheticolumbar block was assessed by comparing local O2 consumption in the ischaemic extremity. It was found that both techniques lead to vasodilation sufficient to normalise local values. Acupuncture, however, has a more protracted effect up to 24 hr., as compared with only 12 hr after pharmacological block.