This study was designed to analyze the effect of vancomycin on the cytoplasmic membrane fatty acid (FA) composition of vancomycin-resistant Staphylococcus aureus (VRSA), vancomycin-intermediate resistant S. aureus (VISA), and vancomycin-susceptible S. aureus. One low-level vancomycin-resistant isolate (LLR-VRSA) termed CP2, along with two vancomycin intermediate-resistant S. aureus isolates (VISA-CP1) and Mu50 (ATCC #700699), were studied. The LLR-VRSA isolate CP2, recovered from the blood sample of a postoperative cardiac patient, exhibited vanA type vancomycin resistance [minimum inhibitory concentration (MIC) 16 μg/ml], and the vanA cassette was located on a plasmid. CP1, isolated from the pus sample of the same patient, exhibited vancomycin intermediate resistance (MIC 8 μg/ml) in the absence of the vanA, vanB, or vanC gene. As susceptible controls, we used PSA (vancomycin MIC 2 μg/ml), which was isolated from the pus sample of a neonate, and S. aureus (ATCC# 29213). Membrane FA analysis was carried out using gas chromatography coupled with mass spectrometry. For this purpose, CP1, CP2, Mu50, and the susceptible control isolates were grown in the presence and absence of vancomycin. Comparative analysis showed an increase in the relative proportion of unsaturated FAs during growth under vancomycin stress. The isolate CP2 (LLR-VRSA) exhibited a higher MIC to vancomycin than the other isolates used in present study (16 μg/ml) and under vancomycin stress conditions, quantitatively, it showed a high rate of conversion of saturated to unsaturated membrane FAs than CP1, Mu50 (VISA isolate) and the susceptible control PSA. The rate of saturated-to-unsaturated FA conversion increased as the concentration of vancomycin in the growth media was increased. Therefore, it is concluded that S. aureus tend to modify their membrane lipid chemistry from saturated to unsaturated in order to survive in a vancomycin stress environment.

A series of first-principles Monte Carlo simulations in the isobaric-isothermal ensemble were carried out for liquid water at ambient conditions (T=298 K and p=1 atm). The Becke-Lee-Yang-Parr (BLYP) exchange and correlation energy functionals and norm-conserving Goedecker-Teter-Hutter (GTH) pseudopotentials were employed with the CP2 K simulation package to examine systems consisting of 64 water molecules. The fluctuations in the system volume encountered in simulations in the isobaric-isothermal ensemble require a reconsideration of the suitability of the typical charge-density cutoff and the regular grid-generation method previously used for the computation of the electrostatic energy in first-principles simulations in the microcanonical or canonical ensembles. In particular, it is noted that a much higher cutoff is needed and that the most computationally efficient method of creating grids can result in poor simulations. Analysis of the simulation trajectories using a very large charge-density cutoff at 1200 Ry and four different grid-generation methods point to a significantly underestimated liquid density of about 0.8 g cm-3 resulting in a somewhat understructured liquid (with a value of about 2.7 for the height of the first peak in the oxygen-oxygen radial distribution function) for BLYP-GTH water at ambient conditions. In addition, a simulation using a charge-density cutoff at 280 Ry yields a higher density of 0.9 g cm-3, showing the sensitivity of the simulation outcome to this parameter.

Two cadmium(ii) coordination polymers (CPs) of compositions {[Cd(H2O)4(4-BPDB)][BPDC]}n (CP1) and {[Cd(H2O)(BrIP)(BTTMB)]·4MeOH}n (CP2) have been synthesized by solvothermal methods and characterized by several analytical methods including SXRD (Single Crystal X-ray Diffraction). The structure of CP1 can be described as a 1D cationic chain, {[Cd(H2O)4(4-BPDB)]2+}n and discrete BPDC counter anions. The structure of CP2 revealed an undulated 2D sql net comprising Cd2+ nodes bridged by the ditopic N-donor, BTTMB and dicarboxylate BrIP involved in μ2-η1η1η1η1 coordination. Supramolecular interactions in both CPs generate 3D hydrogen bonded architectures. The solid state fluorescence properties of these d10 metal ion containing CPs have been investigated. Fluorescence emission of CP1 suspended in acetonitrile is observed to be selectively quenched by acetone (LOD = 0.15 mM) over other common laboratory solvents.

Reactions of K6Sn2Se6 (1) with [Cp*CoCl]2 were investigated in order to probe the stability of the formal +3 oxidation state at Sn and possible ligand properties of heteroatomic zintl-type anion "Sn2Se6(6)- ". From these experiments, we obtained the following compounds that are oxidized to different extent as a result of the reaction with SnIII: [Cp2*Co][Cl2Co(mu2-Cl)2Li(thf)2] (2), [(Cp*Co)3(mu-Se)2] (3), [(Cp*Co)3(mu3-Se)2][Cl2Co(mu2-Cl)2Li(thf)2] (4), and [(Cp*Co)4(mu3-Se)4] (5). These compounds were structurally characterized by single-crystal X-ray diffractometry. It shows that the reaction conditions strongly affect the type and oxidation state of the isolated product. Two of the observed compounds, 3 and 4, are closely related both structurally and electronically; this is discussed and further illustrated by cyclovoltammetric measurements. The choice of the terminal Cp* ligand attached to the transition metal in the reactand complex is assumed to be basically dependent for the alignment of unexpected structural details when compared with known compounds of similar compositions. In conclusion, 1 is observed to act as mild oxidant as well as selenide donor, but is not in the position to keep its Sn-Se framework under the given reaction conditions.

Metalloradical species [Co2 Fv(CO)4 ](.+) (1(.+) , Fv=fulvalenediyl) and [Co2 Cp2 (CO)4 ](.+) (2(.+) , Cp=η(5) -C5 H5 ), formed by one-electron oxidations of piano-stool cobalt carbonyl complexes, can be stabilized with weakly coordinating polyfluoroaluminate anions in the solid state. They feature a supported and an unsupported (i.e. unbridged) cobalt-cobalt three-electron σ bond, respectively, each with a formal bond order of 0.5 (hemi-bond). When Cp is replaced by bulkier Cp* (Cp*=η(5) -C5 Me5 ), an interchange between an unsupported radical [Co2 Cp*2 (CO)4 ](.+) (anti-3(.+) ) and a supported radical [Co2 Cp*2 (μ-CO)2 (CO)2 ](.+) (trans-3(.+) ) is observed in solution, which cocrystallize and exist in the crystal phase. 2(.+) and anti-3(.+) are the first stable thus isolable examples that feature an unsupported metal-metal hemi-bond, and the coexistence of anti-3(.+) and trans-3(.+) in one crystal is unprecedented in the field of dinuclear metalloradical chemistry. The work suggests that more stable metalloradicals of metal-metal hemi-bonds may be accessible by using metal carbonyls together with large and weakly coordinating polyfluoroaluminate anions.

BACKGROUND

Phenotypic and genotypic heterogeneity is a known feature of many cancers. Whether serum tumor marker kinds vary and change following chemotherapy is still unclear. The aim of this study was to investigate whether there is a change in the expression of serum tumor markers following chemotherapy, and the potential clinical significance in patients with epithelial ovarian carcinoma (EOC) or primary serous peritoneal carcinoma (PSPC).

METHODS

Samples were collected before surgery, during chemotherapy and during follow-up for enzyme-linked immunosorbent assay (ELISA)-based evaluation of serum CA-125, CA19-9 and CP2 levels in patients with EOC or PSPC who had received primary debulking surgery followed by adjuvant chemotherapy. In total, 72 patients were examined, including 37 patients with recurrent lesions and 35 patients receiving first-line chemotherapy.

RESULTS

In 35 de novo patients, 20% (7/35) demonstrated a significant changed serum tumor marker kinds among whom the patients with mucinous carcinoma (57.1%, 4/7) showed resistance to chemotherapy. In the 37 recurrent patients, 51.4% (19/37) had changed serum tumor markers, of whom 57.9% (11/19) presented with serous carcinoma. There was no significant difference in median progression-free survival or overall survival in patients with drug-sensitive or drug-resistant recurrence in patients with changed tumor marker kinds relative to those with unchanged markers. However, for patients with changed serum tumor markers there was a trend towards prolonged survival compared with the unchanged serum tumor marker group. In the 17 patients with secondary recurrence, 37.5% (6/17) had changed tumor marker levels. The ratios of CA-125/CP2 and CA-125/CA19-9 were significantly different after either chemotherapy or recurrence.

CONCLUSIONS

Serum tumor marker expression in patients with EOC or PSPC may change after chemotherapy or recurrence, indicating that in addition to the markers that are abnormal before surgery, those markers that are normal should also be monitored during chemotherapy and follow-up.

The binding constants (K) for the interaction of three copigments (CP), two epimeric vinylcatechin dimers (CP1 and CP2), and catechin dimer B3 (CP3) with two pigments, malvidin-3-glucoside (oenin) and malvidin-3,5-diglucoside (malvin), were determined. The K values clearly show that both vinylcatechin dimers have much higher affinity for oenin and malvin than dimer B3: K(CP2)>> K(CP1)>>) K(CP3). Quantum mechanics and molecular dynamics calculations were also performed to interpret the binding data and specify the relative arrangement of the pigment and copigment molecules within the complexes.

The aim of this study is to investigate interactions possibly taking place in red wine between three flavanols (copigments, CP), i.e., two epimeric vinylcatechin dimers (CP1 and CP2) and catechin dimer B3 (CP3), and a specific pigment resulting from the condensation between the main grape anthocyanin malvidin 3-O-glucoside (oenin) and catechin, catechin-(4→8)-oenin. By comparison with our previous work on oenin itself, the influence of the catechin moiety of the anthocyanin in the binding is established. The thermodynamic parameters show that both vinylcatechin dimers exhibit a higher affinity for catechin-(4→8)-oenin, in comparison with proanthocyanidin B3, as previously observed with oenin. However, the corresponding binding constants are weaker, probably due to steric hindrance in the anthocyanin brought by the flavanol nucleus. Consequently, catechin-(4→8)-oenin should be much less stabilized by copigmentation in hydroalcoholic solution than oenin. Quantum mechanics and molecular dynamics simulations are also performed to interpret the binding data, to specify the relative arrangement of the pigment and copigment molecules within the complexes, and to interpret their absorption properties in the visible range.

The physical adsorption and the chemical coupling of recombinant proteins of Trypanosoma cruzi onto polystyrene and core-shell carboxylated particles were respectively investigated with the ultimate aim of producing latex-protein complexes to be used in an immunoagglutination assay able to detect the Chagas disease. To this effect, two single proteins (RP1 and RP5) and a multiepitope protein derived from three antigenic peptides (CP2) were evaluated, and sensitizations were carried out at different pHs. The maximum physical adsorption was produced at pHs close to the protein isoelectric point (i.e., pH 6 for RP5 and pH 5 for RP1 and CP2). High fractions of antigens were chemically bound to the carboxyl groups, and the highest surface density of linked protein was also observed at pHs close to the protein isoelectric point. The three latex-protein complexes obtained by covalent coupling at such pHs were tested with sera from a panel of 16 infected and 16 non-infected patients. In the immunoagglutination assays, the latex-CP2 complex produced the best discrimination between positive and negative sera.

The effect of changes in extracellular Cap2+ concentration (Cp2+) (0.65-5.2 mM) on increases of left ventricular mechanical performance and O2 consumption (MVO2) induced by the elevation perfusion pressure (PP) was studied in closed and drained ventricle isolated rat heart preparation perfused by the modified Langendorff technique. Coronary flow and left ventricular systolic pressure did not depend on Cap2+. In contrast, elevation of Cap2+ potentiated the rise of dp/dtmax considerably. Thebesius flow was significantly lower at high Cap2+ levels. The initial phase of the MVO2 increase in the low PP range (20-60 mmHg) was not influenced by changes in Cap2+. At higher PP levels (80-160 mmHg) elevation of Ca2p+ significantly potentiated the MVO2 increase in the closed ventricle preparation, and if the slowly rising phase of MvO2 increase induced by the elevation of PP was eliminated by draining the left ventricle, the magnitude of the plateau phase of MVO2-PP relationship was a function of Cap2+. Hypoperfusion at the lowest PP level (20 mmHg) (coronary flow was always less than 2 ml/min) caused significant impairment of mechanical activity and MVO2, which could not be compensated for by elevation of Cap2+.

1. Two carboxyl-terminally amidated peptides (CP1 and CP2) were isolated from the blue crab (Callinectes sapidus) eyestalks by a method of carboxyl-terminal analysis. 2. The peptides were sequenced as pGlu-Gly-Arg-Phe-amide (CP1) and pGlu-Leu-Gly-Arg-Phe-amide (CP2). 3. Each carboxyl-terminus of the peptides was precisely determined by amino acid analysis utilizing phenylisothiocyanate derivatives. 4. CP1 was identical to the sea anemone neuropeptide, Antho-RFamide.

Density functional calculations are used to investigate the structure and bonding in several unusual cyclopentadienyl complexes of uranium with nitrogen-containing ligands. The U(VI) imido complex Cp2U(NPh)2 and the U(IV) amido complex Cp2(NHPh)2 are examined and the important orbitals involved in the U-N bonds are analyzed. The recently synthesized 22-electron U(IV) hydrazonato complex U(IV) Cp*2U(Me-N-N=CR2)2 is explored from the standpoint of an expanded valence shell, and the differences between the structures and thermochemistries of U(IV) and Zr(IV) complexes are probed.

Dually responsive diblock random copolymers poly(nPA0.8-co-DEAEMA0.2)-block-poly(nPA0.8-co-EA0.2) were made from N-n-propylacrylamide (nPA), 2-(diethylamino)ethyl methacrylate (DEAEMA) and N-ethylacrylamide (EA) via reversible addition-fragmentation chain transfer (RAFT) polymerization. Copolymers of different block length ratios, poly(nPA28-co-DEAEMA7)-block-poly(nPA29-co-EA7) (P1) and poly(nPA28-co-DEAEMA7)-block-poly(nPA70-co-EA18) (P2), self-assemble into vesicles and micelles, responding to external stimuli in aqueous solutions, and both show "schizophrenic" inversion behavior when the pH and temperature are varied. The relative lengths of the two blocks are shown to affect the self-assembly of amphiphilic diblock copolymers. P1 has a similar length for both blocks and forms spherical vesicles with the first block poly(nPA29-co-EA7) as the membrane inner layer at pH 7 and 37 °C (above the cloud point of the more hydrophobic block, CP1), while spherical micelle-like aggregates are obtained at pH 10 and 25 °C (above CP1) with the second block poly(nPA28-co-DEAEMA7) as the core. In comparison, P2 has a different block length ratio (1 : 3, thus a much longer second block) and forms spherical micelles above CP1 at both pH 7 (the second block as the core) and pH 10 (the first block as the core). Further aggregation was observed by heating the polymer solution above the cloud point of the more hydrophilic block (CP2). The variation of the length and chemical composition of the blocks allows the tuning of the responsiveness of the block copolymers toward both pH and temperature and determines the formation of either micelles or vesicles during the aggregation.

The Cryptosporidium parvum T7 phage display library was screened by using Caco-2 cells. Five specific gene fragments were identified by blasting sequences in GenBank, one of which encoding the CP2 protein was previously identified as a surface molecule of sporozoites and involved in parasite invasion. The others are hypothetic proteins with unknown functions. Bioinformatic analysis of these proteins indicated that they may be involved in the host-parasite interactions.

An ultrasensitive electrochemical genosensor has been fabricated for the double determination of two different specific sequences deduced from the maternally expressed gene3 (MEG3) lncRNA (long noncoding RNA), which was demonstrated by coupling RNase A-aided target recycling with DNA supersandwich-induced signal enhancement, based on a composite interface of graphene-like tungsten disulfide/dendritic gold nanostructures (WS2/DGN). Firstly, duple target sequences of T1 and T2 were captured by the primer probes of P1/P2 functionalized Fe3O4@C magnetic nanoparticles, via the DNA/RNA hybridization between T1/T2 and P1/P2. In the presence of RNase A, T1 and T2 were released to trigger the target recycling, accompanied by the generation of numerous intermediate DNAs designated as IT1 and IT2, respectively. After the magnetic separation, the IT1 and IT2 were liberated and hybridized with the capture probes of CP1/CP2 loaded DGN/WS2 modified electrode. Subsequently, the stepwise DNA hybridization chain reactions (HCR) labeled with ferrocene (Fc) and methyleneblue (MB) were processed, respectively. The DPV current values of Fc and MB were recorded, which were proportional with the concentration of T1 and T2, respectively. Using the multiplexed amplification strategy, this newly designed genosensor provided a wide linear range from 1 fM to 100 pM with a low detection limit of 0.25 fM for T1 and 0.3 fM for T2. The application of the genosensor in real serum sample has also been studied, confirming the excellent selectivity and sensitivity for the application in bioanalysis and clinical diagnostics.

The diphosphaazide complex (Mes*NPP)Nb(N[Np]Ar)3 (Mes* = 2,4,6-tri-tert-butylphenyl, Np = neopentyl, Ar = 3,5-Me2C6H3), 1, has previously been reported to lose the P2 unit upon gentle heating, to form (Mes*N)Nb(N[Np]Ar)3, 2. The first-order activation parameters for this process have been estimated here using an Eyring analysis to have the values Delta H(double dagger) = 19.6(2) kcal/mol and Delta S(double dagger) = -14.2(5) eu. The eliminated P2 unit can be transferred to the terminal phosphide complexes P[triple bond]M(N[(i)Pr]Ar)3, 3-M (M = Mo, W), and [P[triple bond]Nb(N[Np]Ar)3](-), 3-Nb, to give the cyclo-P3 complexes (P3)M(N[(i)Pr]Ar)3 and [(P3)Nb(N[Np]Ar)3](-). These reactions represent the formal addition of a P[triple bond]P triple bond across a M[triple bond]P triple bond and are the first efficient transfers of the P2 unit to substrates present in stoichiometric quantities. The related complex (OC)5W(Mes*NPP)Nb(N[Np]Ar)3, 1-W(CO)5, was used to transfer the (P2)W(CO)5 unit in an analogous manner to the substrates 3-M (M = Mo, W, Nb) as well as to [(OC)5WP[triple bond]Nb(N[Np]Ar)3](-). The rate constants for the fragmentation of 1 and 1-W(CO)5 were unchanged in the presence of the terminal phosphide 3-Mo, supporting the hypothesis that molecular P2 and (P2)W(CO)5, respectively, are reactive intermediates. In a reaction related to the combination of P[triple bond]P and M[triple bond]P triple bonds, the phosphaalkyne AdC[triple bond]P (Ad = 1-adamantyl) was observed to react with 3-Mo to generate the cyclo-CP2 complex (AdCP2)Mo(N[(i)Pr]Ar)3. Reactions of the electrophiles Ph3SnCl, Mes*NPCl, and AdC(O)Cl with the anionic, nucleophilic complexes [(OC)5W(P3)Nb(N[Np]Ar)3](-) and [{(OC)5W}2(P3)Nb(N[Np]Ar)3](-) yielded coordinated eta(2)-triphosphirene ligands. The Mes*NPW(CO)5 group of one such product engages in a fluxional ring-migration process, according to NMR spectroscopic data. The structures of (OC)5W(P3)W(N[(i)Pr]Ar)3, [(Et2O)Na][{(OC)5W}2(P3)Nb(N[Np]Ar)3], (AdCP2)Mo(N[(i)Pr]Ar)3, (OC)5W(Ph3SnP3)Nb(N[Np]Ar)3, Mes*NP(W(CO)5)P3Nb(N[Np]Ar)3, and {(OC)5W}2AdC(O)P3Nb(N[Np]Ar)3, as determined by X-ray crystallography, are discussed in detail.

Auditory evoked potentials (AEPs) to motion onset in humans are dominated by a fronto-central complex, with a change-negative deflection 1 (cN1) and a change-positive deflection 2 (cP2) component. Here the contribution of veridical motion detectors to motion-onset AEPs was investigated with the hypothesis that direction-specific adaptation effects would indicate the contribution of such motion detectors. AEPs were recorded from 33 electroencephalographic channels to the test stimulus, i.e. motion onset of horizontal virtual auditory motion (60° per s) from straight ahead to the left. AEPs were compared in two experiments for three conditions, which differed in their history prior to the motion-onset test stimulus: (i) without motion history (Baseline), (ii) with motion history in the same direction as the test stimulus (Adaptation Same), and (iii) a reference condition with auditory history. For Experiment 1, condition (iii) comprised motion in the opposite direction (Adaptation Opposite). For Experiment 2, a noise in the absence of coherent motion (Matched Noise) was used as the reference condition. In Experiment 1, the amplitude difference cP2 - cN1 obtained for Adaptation Same was significantly smaller than for Baseline and Adaptation Opposite. In Experiment 2, it was significantly smaller than for Matched Noise. Adaptation effects were absent for cN1 and cP2 latencies. These findings demonstrate direction-specific adaptation of the motion-onset AEP. This suggests that veridical auditory motion detectors contribute to the motion-onset AEP.

Thirty children with cerebral palsy (CP) and 22 typical developing (TD) were tested with 3D-gait analysis. At turning, trunk rotation was larger in CP2 (GMFCS II) than in TD and CP1 (GMFCS I), and head flexion was larger in CP3 (GMFCS III) than TD. Maximum head and trunk flexion values during the entire trial were larger in CP3 than in the other groups, and trunk flexion was larger in CP2 than in TD. Trial time increased with GMFCS-level. Less trunk rotation than TD and CP1 reflects spatial insecurity in CP2, which in CP3 is compensated by the walker. The flexed head and trunk in CP3 and trunk in CP2 may reflect deficits in proprioception and sensation requiring visual control of the lower limbs.

We study a general model of isotropic two-dimensional spin-1 magnet, which is relevant for the physics of ultracold atoms with hyperfine S=1 spins in an optical lattice at odd filling. We demonstrate a novel mechanism of soliton pairing occurring in the vicinity of a special point with an enhanced SU(3) symmetry: upon perturbing the SU(3) symmetry, solitons with odd CP2 topological charge are confined into pairs that remain stable objects.

There are approximately 20 known species of the genus Cryptosporidium, and among these, 8 infect immunocompetent or immunocompromised humans. C. hominis and C. parvum most commonly infect humans. Differentiating between them is important for evaluating potential sources of infection. We report here the development of a simple and accurate real-time PCR-based restriction fragment length polymorphism (RFLP) method to distinguish between C. parvum and C. hominis. Using the CP2 gene as the target, we found that both Cryptosporidium species yielded 224 bp products. In the subsequent RFLP method using TaqI, 2 bands (99 and 125 bp) specific to C. hominis were detected. Using this method, we detected C. hominis infection in 1 of 21 patients with diarrhea, suggesting that this method could facilitate the detection of C. hominis infections.

White spot syndrome virus (WSSV) and Taura syndrome virus (TSV) are highly pathogenic to penaeid shrimp and have caused significant economic losses in the shrimp culture industry around the world. During 2010 and 2011, both WSSV and TSV were found in Saudi Arabia, where they caused severe mortalities in cultured Indian white shrimp Penaeus indicus. Most outbreaks of shrimp viruses in production facilities can be traced to the importation of infected stocks or commodity shrimp. In an attempt to determine the origins of these viral outbreaks in Saudi Arabia, we performed variable number of tandem repeat (VNTR) analyses for WSSV isolates and a phylogenetic analysis for TSV isolates. From the WSSV genome, the VNTR in open reading frames (ORFs) 125 and 94 were investigated with PCR followed by DNA sequence analysis. The genotypes were categorized as {N125, N94} where N is the number of repeat units in a specific ORF, and the subscript indicates the ORF (i.e. ORFs 125 and 94 in this case). From 15 Saudi Arabia WSSV isolates, we detected 3 genotypes: {6125, 794}, {7125, del94}, and {8125, 1394}. The WSSV genotype of {7125, del94} appears to be a new variant with a 1522 bp deletion encompassing complete coding regions of ORF 94 and ORF 95 and the first 82 bp of ORF 93. For TSV genotyping, we used a phylogenetic analysis based on the amino acid sequence of TSV capsid protein 2 (CP2). We analyzed 8 Saudi Arabian isolates in addition to 36 isolates from other areas: SE Asia, Mexico, Venezuela and Belize. The Saudi Arabian TSV clustered into a new, distinct group. Based on these genotyping analyses, new WSSV and TSV genotypes were found in Saudi Arabia. The data suggest that they have come from wild shrimp Penaeus indicus from the Red Sea that are used for broodstock.

The Dictyostelium mutant HI4 progresses through morphogenesis normally, but is defective in the reverse program of dedifferentiation. In contrast to dedifferentiating wild-type cells, HI4 cells retain the capacity to rapidly reaggregate well after the "erasure event" employing a nonchemotactic aggregation mechanism involving random collisions and cohesion. They also do not lose contact sites A (gp80) at the prescribed time in the dedifferentiation program. HI4 cells accumulate transcripts of the cysteine protease gene CP2 (formerly referred to as 16G1) and the cohesion glycoprotein gene gp80 at the correct times in the morphogenetic program, but abnormally retain these transcripts at high levels well after the prescribed times at which they are lost in wild-type cells during the reverse program of dedifferentiation. The retention of these mRNAs in HI4 cells after the erasure event is not due to abnormal maintenance of a high level of intracellular cAMP during dedifferentiation. The rapid reduction in the level of gp80 transcript which can be effected by the addition of cAMP prior to the erasure event in wild-type cells is also retained by HI4 cells well after the erasure event. The results suggest that cells possess at least two mechanisms for the reduction of gp80 transcript. One involves the immediate response to cAMP and may function during the forward program of development. The second functions specifically during the reverse program of dedifferentiation. It is this latter, erasure-specific mechanism which is selectively defective in the HI4 variant.

BACKGROUND

Even though metal ceramic restorations (MCRs) are widely used by clinicians, the influence of the metal on the color of overlaying porcelain is unknown.

OBJECTIVE

The purpose of this study was to analyze the color alterations of different types of metal ceramic alloys during several stages of metal surface preparation and to determine the effect of those changes on the resulting color of opaque porcelain (OP).

METHODS

Seven different types of alloys (3 base metal, 3 noble, and 1 high noble) were used to prepare disk-shaped specimens (1 mm × 10 mm, n=3), followed by OP application (0.1 mm). L*a*b* values of specimens were recorded after different stages of metal surface preparation (ingot, after casting, after oxidation, and after the OP application) in addition to the shade tab of OP B1 (target shade). L*a*b* values of alloys were measured from the ingot structure to the OP application stage and statistically analyzed (Repeated measures ANOVA, and Bonferroni corrected paired t test, α=.05). L*a*b* values of OP applied groups and the OP shade tab (target shade) were analyzed (1-way ANOVA with Dunnett's multiple comparison test, α=.05). The color differences of the target shade both before and after OP application were calculated and statistically analyzed (1-way ANOVA, Ryan-Einot-Gabriel-Welsch Multiple Range Test, α=.05).

RESULTS

The L* values of all alloys changed significantly after each stage except for 2 alloys (V-Deltaloy SF (N-VDSF)) and (Gnathos Plus (HN-GP)) after casting and airborne-particle abrasion (P<.05). The a* value of all alloys increased after casting. Changes in the a* coordinate were significant except for one of the base metal alloys (P<.05). The a* coordinate changes of alloys showed variation in direction after oxidation and OP application (P<.05). The b* coordinate changes of alloys showed variation in direction after each stage (P<.05). The L*a*b* values of some OP applied alloys were significantly different from that of the OP shade tab (P<.05). Color difference values (ΔE (OP applied alloy-target shade)) of 2 OP-applied alloys (Cerapall 2 (N-CP2) and Ceradelta (N-CD)) were significantly different (P<.05) and higher than the other OP-applied alloys.

CONCLUSIONS

The achromatic color behavior of different alloys was all in the same direction at all metal surface preparation stages. The chromatic behavior of the different alloys was primarily towards the same direction after casting and airborne-particle abrasion, whereas it varied after oxidation and OP application. The color difference of OP for all alloys, regardless of their type, was not visually perceivable when compared to the target shade (ΔE<2.6).

Cryptosporidiosis, a gastrointestinal disease caused by protozoans of the genus Cryptosporidium, is a common cause of diarrheal diseases and often fatal in immunocompromised individuals. Bifunctional thymidylate synthase-dihydrofolate reductase (TS-DHFR) from Cryptosporidium hominis (C. hominis) has been a molecular target for inhibitor design. C. hominis TS-DHFR inhibitors with nM potency at a biochemical level have been developed however drug delivery to achieve comparable antiparasitic activity in Cryptosporidium infected cell culture has been a major hurdle for designing effective therapies. Previous mechanistic and structural studies have identified compound 906 as a nM C. hominis TS-DHFR inhibitor in vitro, having μM antiparasitic activity in cell culture. In this work, proof of concept studies are presented using a nanotherapy approach to improve drug delivery and the antiparasitic activity of 906 in cell culture. We utilized PLGA nanoparticles that were loaded with 906 (NP-906) and conjugated with antibodies to the Cryptosporidium specific protein, CP2, on the nanoparticle surface in order to specifically target the parasite. Our results indicate that CP2 labeled NP-906 (CP2-NP-906) reduces the level of parasites by 200-fold in cell culture, while NP-906 resulted in 4.4-fold decrease. Moreover, the anticryptosporidial potency of 906 improved 15 to 78-fold confirming the utility of the antibody conjugated nanoparticles as an effective drug delivery strategy.