The days ahead

How are government controls affecting healthcare foodservices?Are growing elderly & outpatient populations changing foodservices' missions. What are the hottest trends? As members of the American Society for Hospital Food Service Administrators prepared for their annual meeting in Orlando, FM queried 11 Southeastern directors to learn their most pressing concerns & how they are positioning their departments for the future.     

Using multiwalled carbon nanotube modified electrodes for the adsorptive striping voltammetric determination of hesperidin

Hesperidin, a flavone glycoside found in the skins and juices of citrus fruits, can be detected using multiwalled carbon nanotube (MWCNT)-modified electrodes using the technique of adsorptive stripping voltammetry (AdSV) with accumulation at open circuit potential. This is relevant because hesperidin can be used as an indication of the citrus fruit juice's freshness. The oxidation mechanism to explain the observed voltammetry corresponds to the redox chemistry of the guaiacol sub-unit within the hesperidin molecular structure. Hesperidin could be detected over a linear range up to 30 μM, and with a detection limit of 0.61 μM and 7 nM, with less than 5% variation between different electrodes, using cyclic voltammetric or square wave adsorptive stripping techniques respectively. This methodology was extended to MWCNT-modified screen-printed electrodes (MWCNT-SPEs), allowing the development of a cheap, mass produced, disposable sensor that we show is capable of measuring the concentration of hesperidin in real orange juice samples, and be applied within the citrus fruit industry.     

Continuous dense‐phase CO2 processing of a coconut water beverage

Effects of dense‐phase CO₂ (DPCD) on microbial, physical, chemical and sensorial quality of coconut water (CW) beverage were evaluated. Pressure during DPCD treatment was not significant in microbial reduction whereas temperature and % CO₂ levels were significant. DPCD‐treated (34.5 MPa, 25 °C, 13% CO₂, 6 min), heat‐pasteurised (74 °C, 15 s) and untreated CW beverages were evaluated during 9 weeks of refrigerated storage (4 °C). Total aerobic bacteria of DPCD and heat‐treated samples decreased whereas that of untreated samples increased to >10⁵ CFU mL^?1 after 9 weeks. DPCD increased titratable acidity but did not change pH (4.20) and °Brix (6.0). Likeability of DPCD‐treated CW was similar to untreated. Heat‐treated samples were less liked (α = 0.05) at the beginning of storage. Off flavour and taste‐difference‐from‐control scores of heated samples were higher than DPCD during the first two weeks. DPCD extended shelf life of acidified, sweetened and carbonated CW over 9 weeks at 4 °C.     

Synthesis and biological properties of novel pyridinioalkanoyl thiolesters (PATE) as anti-HIV-1 agents that target the viral nucleocapsid protein zinc fingers

Nucleocapsid p7 protein (NCp7) zinc finger domains of the human immunodeficiency virus type 1 (HIV-1) are being developed as antiviral targets due to their key roles in viral replication and their mutationally nonpermissive nature. On the basis of our experience with symmetrical disulfide benzamides (DIBAs; Rice et al. Science 1995, 270, 1194-1197), we synthesized and evaluated variants of these dimers, including sets of 4,4'- and 3,3'-disubstituted diphenyl sulfones and their monomeric benzisothiazolone derivatives (BITA). BITAs generally exhibited diminished antiviral potency when compared to their disulfide precursors. Novel, monomeric structures were created by linking haloalkanoyl groups to the benzamide ring through -NH-C(=O)- (amide) or -S-C(=O)- (thiolester) bridges. Amide-linked compounds generally lacked antiviral activity, while haloalkanoyl thiolesters and non-halogen-bearing analogues frequently exhibited acceptable antiviral potency, thus establishing thiolester benzamides per se as a new anti-HIV chemotype. Pyridinioalkanoyl thiolesters (PATEs) exhibited superior anti-HIV-1 activity with minimal cellular toxicity and appreciable water solubility. PATEs were shown to preferentially target the NCp7 Zn finger when tested against other molecular targets, thus identifying thiolester benzamides, and PATEs in particular, as novel NCp7 Zn finger inhibitors for in vivo studies.     

Exploring structure-activity relationships around the phosphomannose isomerase inhibitor AF14049 via combinatorial synthesis

Phosphomannose Isomerase (PMI) has been shown by genetic methods to be an essential enzyme in fungal cell wall biosynthesis. The PMI inhibitor AF14049 was discovered as an unanticipated side product from high-throughput library screening against the enzyme from C, albicans. Solid-phase synthetic methods were developed and a series of libraries and discrete analogs synthesized to explore SAR around AF14049.     

Transformation of hematopoietic cell lines to growth-factor independence and induction of a fatal myelo- and lymphoproliferative disease in mice by retrovirally transduced TEL/JAK2 fusion genes

Recent reports have demonstrated fusion of the TEL gene on 12p13 to the JAK2 gene on 9p24 in human leukemias. Three variants have been identified that fuse the TEL pointed (PNT) domain to (i) the JAK2 JH1-kinase domain, (ii) part of and (iii) all of the JH2 pseudokinase domain. We report that all of the human TEL/JAK2 variants, and a human/mouse chimeric hTEL/mJAK2(JH1) fusion gene, transform the interleukin-3 (IL-3)-dependent murine hematopoietic cell line Ba/F3 to IL-3-independent growth. Transformation requires both the TEL PNT domain and JAK2 kinase activity. Furthermore, all TEL/JAK2 variants strongly activated STAT 5 by phosphotyrosine Western blots and by electrophoretic mobility shift assays (EMSA). Mice (n = 40) transplanted with bone marrow infected with the MSCV retrovirus containing either the hTEL/mJAK2(JH1) fusion or its human counterpart developed a fatal mixed myeloproliferative and T-cell lymphoproliferative disorder with a latency of 2-10 weeks. In contrast, mice transplanted with a TEL/JAK2 mutant lacking the TEL PNT domain (n = 10) or a kinase-inactive TEL/JAK2(JH1) mutant (n = 10) did not develop the disease. We conclude that all human TEL/JAK2 fusion variants are oncoproteins in vitro that strongly activate STAT 5, and cause lethal myelo- and lymphoproliferative syndromes in murine bone marrow transplant models of leukemia.     

Identification of residues in the cysteine-rich domain of Raf-1 that control Ras binding and Raf-1 activity

We have identified mutations in Raf-1 that increase binding to Ras. The mutations were identified making use of three mutant forms of Ras that have reduced Raf-1 binding (Winkler, D. G., Johnson, J. C., Cooper, J. A., and Vojtek, A. B. (1997) J. Biol. Chem. 272, 24402-24409). One mutation in Raf-1, N64L, suppresses the Ras mutant R41Q but not other Ras mutants, suggesting that this mutation structurally complements the Ras R41Q mutation. Missense substitutions of residues 143 and 144 in the Raf-1 cysteine-rich domain were isolated multiple times. These Raf-1 mutants, R143Q, R143W, and K144E, were general suppressors of three different Ras mutants and had increased interaction with non-mutant Ras. Each was slightly activated relative to wild-type Raf-1 in a transformation assay. In addition, two mutants, R143W and K144E, were active when tested for induction of germinal vesicle breakdown in Xenopus oocytes. Interestingly, all three cysteine-rich domain mutations reduced the ability of the Raf-1 N-terminal regulatory region to inhibit Xenopus oocyte germinal vesicle breakdown induced by the C-terminal catalytic region of Raf-1. We propose that a direct or indirect regulatory interaction between the N- and C-terminal regions of Raf-1 is reduced by the R143W, R143Q, and K144E mutations, thereby increasing access to the Ras-binding regions of Raf-1 and increasing Raf-1 activity.