Three-Component Synthesis of Pyridylacetic Acid Derivatives by Arylation/Decarboxylative Substitution of Meldrum's Acids.

A convenient and simple three-component synthesis of substituted pyridylacetic acid derivatives is reported. The approach centers on the dual reactivity of Meldrum's acid derivatives, initially as nucleophiles to perform substitution on activated pyridine-N-oxides, then as electrophiles with a range of nucleophiles to trigger ring-opening and decarboxylation.

Direct sulfonylation of anilines mediated by visible light.

Sulfones feature prominently in biologically active molecules and are key functional groups for organic synthesis. We report a mild, photoredox-catalyzed reaction for sulfonylation of aniline derivatives with sulfinate salts, and demonstrate the utility of the method by the late-stage functionalization of drugs. Key features of the method are the straightforward generation of sulfonyl radicals from bench-stable sulfinate salts and the use of simple aniline derivatives as convenient readily available coupling partners.

C-H Cyanation of 6-Ring N-Containing Heteroaromatics.

Heteroaromatic nitriles are important compounds in drug discovery, both for their prevalence in the clinic and due to the diverse range of transformations they can undergo. As such, efficient and reliable methods to access them have the potential for far-reaching impact across synthetic chemistry and the biomedical sciences. Herein, we report an approach to heteroaromatic C-H cyanation through triflic anhydride activation, nucleophilic addition of cyanide, followed by elimination of trifluoromethanesulfinate to regenerate the cyanated heteroaromatic ring. This one-pot protocol is simple to perform, is applicable to a broad range of decorated 6-ring N-containing heterocycles, and has been shown to be suitable for late-stage functionalization of complex drug-like architectures.

Precious-Metal-Free Heteroarylation of Azlactones: Direct Synthesis of α-Pyridyl, α-Substituted Amino Acid Derivatives.

A one-pot, three-component synthesis of α-pyridyl, α-substituted amino acid derivatives is described. The key transformation is a direct, precious-metal-free heteroarylation of readily available, amino acid derived azlactones with electrophilically activated pyridine N-oxides. The resulting intermediates can be used directly as efficient acylating agents for a range of nucleophiles, leading to the heteroarylated amino acid derivatives in a single vessel.

A convergent, umpoled synthesis of 2-(1-amidoalkyl)pyridines.

A convenient, one-pot, two-component synthesis of 2-(1-amidoalkyl)pyridines is reported, based upon the substitution of suitably-activated pyridine N-oxides by azlactone nucleophiles, followed by decarboxylative azlactone ring-opening. The synthesis obviates the need for precious metal catalysts to achieve a formal enolate arylation reaction, and constitutes a formally 'umpoled' approach to this valuable class of bioactive structures.

Application of ruthenium complexes of triazole-containing tridentate ligands to asymmetric transfer hydrogenation of ketones.

The synthesis of a series of tridentate ligands based on a homochiral 1,2-diamine structure attached to a triazole group and their subsequent applications to the asymmetric transfer hydrogenation of ketones are described. In the best cases, alcohols of up to 93% ee were obtained. Although base is not required, the use of Ru(3)(CO)(12) as metal source is essential, indicating a unique mechanism for the formation of the active catalyst.

Developing asymmetric iron and ruthenium-based cyclone complexes; complex factors influence the asymmetric induction in the transfer hydrogenation of ketones.

The preparation of a range of asymmetric iron and ruthenium-cyclone complexes, and their application to the asymmetric reduction of a ketone, are described. The enantioselectivity of ketone reduction is influenced by a single chiral centre in the catalyst, as well as by the planar chirality in the catalyst. This represents the first example of asymmetric ketone reduction using an iron cyclone catalyst.

Hydrogen generation from formic acid and alcohols using homogeneous catalysts.

This tutorial review describes recent progress in the development of homogeneous catalytic methodology for the direct generation of hydrogen gas from formic acid and alcohols.

Hepatitis B virus X protein differentially activates RAS-RAF-MAPK and JNK pathways in X-transforming versus non-transforming AML12 hepatocytes.

The hepatitis B virus (HBV) X protein (pX) is implicated in hepatocarcinogenesis of chronic HBV patients by an unknown mechanism. Activities of pX likely relevant to hepatocyte transformation include activation of the mitogenic RAS-RAF-MAPK and JNK pathways. To assess the importance of mitogenic pathway activation by pX in transformation, we employed a cellular model system composed of two tetracycline-regulated, pX-expressing cell lines, constructed in AML12-immortalized hepatocytes. This system includes the differentiated 3pX-1 and the de-differentiated 4pX-1 hepatocytes. Our studies have demonstrated that conditional pX expression transforms only 3pX-1 cells. Here, comparative in vitro kinase assays and various in vivo analyses demonstrate that pX affects an inverse activation of RAS-RAF-MAPK and JNK pathways in 3pX-1 versus 4pX-1 cells. Sustained pX-dependent RAS-RAF-MAPK pathway activation is observed in pX-transforming 3pX-1 cells, whereas sustained pX-dependent JNK pathway activation is observed in pX non-transforming 4pX-1 cells. This differential, pX-dependent mitogenic pathway activation affects differential activation of cAMP-response element-binding protein and c-Jun and determines the proliferative response of 3pX-1 and 4pX-1 cells. Furthermore, tetracycline-regulated, pX-NLS-expressing cell lines demonstrate that expression of the nuclear pX-NLS variant minimally activates the RAS-RAF-MAPK pathway and results in markedly reduced transformation. These results link sustained, pX-mediated activation of RAS-RAF-MAPK pathway to hepatocyte transformation.

Effects of fescue and clover forage on serum lactate dehydrogenase and glucose 6-phosphate dehydrogenase isoenzymic profiles in steers.

We determined the effects of forage type on isoenzymes of lactate dehydrogenase (LDH) and glucose 6-phosphate dehydrogenase (G6PDH). Forty-eight crossbred steers were randomly allotted to replicated pastures consisting of fungus-infected (Neotyphodium coenophialum) fescue or fungus-free fescue each with or without ladino clover overseeding. At the end of the 180-d grazing period, serum was harvested from the steers. Steers were finished in a feedlot and slaughtered after approximately 150 d in the feedlot. Isoenzymes for LDH and G6PDH were separated using PAGE. Five LDH isoenzymes (L1-15) were typically detected. Isoenzyme L1 (most anodic) had the greatest area percent as detected by laser densitometry (72, 12, 10, 5, and 7%, respectively, for L1, L2, L3, L4, and L5). Four proteins had G6PDH activity (G1-G4) with G2 having the greatest area percent (15, 52, 27, and 14, respectively, for G1, G2, G3, and G4). Isoenzymes within a dehydrogenase were correlated (P < .05). In addition, area percentage of L1 was correlated (P < .05; r = .34) with area percentage of G2, and area percentage of L4 was correlated (P < .07; r = .73) with area percentage of G1. Area percentages of L1, L2, and L3 were affected by an interaction (P < .09) of forage types. Body weight gains for steers grazing endophyte-infected fescue were depressed (P < .05); however, steers compensated with increased (P < .05) weight gains during the finishing phase. Fungal toxins produced by Neotyphodium coenophialum may alter an animal's metabolism, growth, and development via shifts in reducing equivalents (NADH).

Differential immediate early gene expression in conditional hepatitis B virus pX-transforming versus nontransforming hepatocyte cell lines.

We report construction and characterization of tetracycline-controlled hepatitis B virus pX-expressing hepatocyte (AML12) cell lines. These cell lines were constructed in AML12 clonal isolates (clones 3 and 4), which express constitutively the tetracycline-controlled transactivator. Since pX is implicated in HCC, this immortalized hepatocyte model system was used to investigate the mechanism of pX in transformation. Clonal isolates of 3pX and 4pX lineages display conditional synthesis of pX mRNA and protein and a 2-fold increase in growth saturation density following tetracycline removal, implicating pX in monolayer overgrowth. Interestingly, only 3pX clones display pX-dependent anchorage independence. Clone 3 lineages express hepatocyte nuclear factor-1alpha and hepatocyte-specific marker genes; clone 4 lineages express hepatocyte nuclear factor-1beta and reduced levels of hepatocyte-specific marker genes, suggesting the importance of the differentiated hepatocyte in pX-mediated oncogenic transformation. Importantly, 3pX and 4pX lineages display differential expression of immediate early genes c-fos and ATF3. The pX-transforming 3pX lineage displays early, pX-dependent induction of ATF3 and prolonged induction of c-fos. The nontransforming 4pX cells display an absence of pX-dependent ATF3 induction and transient induction of c-fos. Our results support the direct link of pX expression to oncogenic transformation in 3pX lineage clones and underscore the advantage of this conditional cellular model system for studying mechanisms of pX-mediated oncogenesis.

Effects of 6-N-propyl-2-thiouracil on growth, hormonal profiles, carcass and reproductive traits of boars.

Neonatal 6-N-propyl-2-thiouracil (PTU)-induced hypothyroidism reduces body weight but increases testicular size in adult male rodents. The objective of this study was to determine the effect of prepubertal PTU treatment on boars. For Experiment I, boars (n = 28) were randomly allotted to eight pens. Each pen received one of four PTU doses (0, 0.01, 0.03 and 0.1% in a basal diet) between 28 and 56 days of age (DOA). Due to a lack of difference among three PTU treatments, PTU-treated boars were pooled. Boars treated with PTU had lower (P < 0.05) ADG during treatment, lighter (P < 0.05) BW after 56 DOA and less (P < 0.05) developed epididymides at 154 DOA. For Experiment II, boars (n = 19) were randomly allotted to six pens. Each pen received one of three PTU treatments orally as: control (carrier), PTU-I (0.002% BW of PTU daily between 7 and 70 DOA), or PTU-II (0.002% BW of PTU daily between 28 and 91 DOA). During treatment, PTU-treated boars had lower (P < 0.05) serum T4 levels, rectal temperature, feed intake and ADG. Boars treated with PTU had lower (P < 0.05) BW between 63 and 154 DOA but higher (P < 0.05) gain/feed between 105 and 133 DOA. Boars treated with PTU had less (P < 0.05) developed epididymides and sperm count per gram testis at 238 DOA. These results suggest that prepubertal PTU-induced hypothyroidism had significant effects on growth, hormonal profiles, and reproductive traits of boars; however, it does not appear to be an effective method for increasing testis size and sperm production of commercial boars.

Polarization changing and beam profile deformation of light during the isotropic Raman-nath acousto-optic interaction.

A spatial Fourier transform approach is used to study the phenomena of polarization changing and beam profile deformation of light during the Raman-Nath, acousto-optic interaction in isotropic media. Starting from the vector version of the well-known Raman-Nath interaction equation and using a spatial Fourier transform allows analytic solutions that encompass the effects of polarization changing and beam-profile deformation for the multiple scattered light to be found in the spatial-frequency domain. Two kinds of sound wave, longitudinal and shear, are assumed to be interacted with the light, whose transverse spatial profile and state of polarization are arbitrary. It is shown that, for light with an arbitrary spatial profile after interaction with the sound wave in the Raman-Nath regime, the spatial profiles of the scattered light are almost the same shape as those of the input light. For the polarization changing part, it is found that the state of polarization and the direction of rotation can alter, depending not only on the sound amplitude but also on the propagation mode of the sound wave. Simulation results are provided to confirm the validity of this approach.

The effect of 5,8,11,14-eicosatetraynoic acid on endothelial cell gene expression.

The endothelium plays a key role in inflammation, hemostasis and organ rejection. We report here that a synthetic polyunsaturated fatty acid, 5,8,11,14-eicosatetraynoic acid (ETYA), selectively inhibits the up-regulation of several genes on endothelial cells. ETYA suppresses endothelial cell activation by inhibiting the up-regulation of adhesion molecules like E-selectin. A runoff assay for E-selectin demonstrated that the suppression is at the level of transcription. The fact that ETYA inhibits E-selectin upon stimulation with a diverse group of stimuli like lipopolysaccharide, tumor necrosis factor-alpha or phorbol 12-myristate 13-acetate, suggests that ETYA does not exert its effect by modifying membrane-bound receptors. The messenger RNA for interleukin-8 and glyceraldehyde phosphate dehydrogenase are not affected. Pre-treatment of endothelial cells with ETYA also prevents the adherence of monocytes to tumor necrosis factor-alpha-stimulated cells.

Selective suppression of endothelial cell activation by arachidonic acid.

Endothelial cell (EC) activation plays a key role in inflammation, thrombosis and organ rejection. Normally, EC are in a quiescent state in which their function is to prevent coagulation and thrombosis, and to participate in the regulation of leukocyte migration from the bloodstream into the tissue. Upon activation with cytokines or other stimuli, EC up-regulate a number of genes, including E-selectin (ELAM-1), intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1, interleukin (IL)-1, IL-8, tissue factor (TF), plasminogen activator inhibitor-1 (PAI-1), MCP-1 (monocyte chemoattractant protein-1) and endothelial cell inducible gene (ECI-6). Arachidonic acid (AA) is produced by several cell types, including EC, and acts on various cells. We report here that AA inhibits the up-regulation of some, but not all genes that are induced with EC activation in a dose-dependent manner. AA suppresses TNF-alpha, IL-1 alpha, LPS or PMA-induced E-selectin expression, as well as mRNA accumulation of E-selectin, ICAM-1 and IL-8 stimulated by TNF-alpha. The inhibition appears to be at the level of transcription. At the same time under the same conditions AA does not, repress mRNA accumulation for PAI-1, ECI-6, MCP-1 and VCAM-1. We suggest that the induced expression of AA with EC activation may result in a negative feedback loop regulating further activation.

Synchronous bilateral carcinoma of the ureter in association with unilateral incomplete duplication of the ureter.

A case of bilateral synchronous ureteral tumors, in association with a unilateral incomplete duplication of ureter, is presented. This patient underwent right nephroureterectomy with removal of a cuff of bladder, partial resection of left ureter, and ileal loop interposition between renal pelvis and bladder. Followup showed no recurrence in the residual urinary tract 2-years postoperatively.

Identification and upregulation of galactose/N-acetylgalactosamine macrophage lectin in rat cardiac allografts with arteriosclerosis.

Using differential mRNA display to uncover potential mediators associated with chronic rejection, we identified a cDNA fragment induced in Lewis to F344 rat cardiac allografts with arteriosclerosis but not Lewis syngrafts. The full-length cDNA (1.4 kb) isolated from a rat cardiac allograft cDNA library was 99% identical to galactose/N-acetylgalactosamine (Gal/GalNAc) macrophage lectin, a cell-surface receptor. This cDNA hybridized in Northern analysis with total RNA from eight cardiac allografts but not with host hearts, syngrafts, or other organs. There was a significant allograft-specific increase in transcript levels measured by reverse transcriptase PCR at days 7, 14, 28, and 75 in comparison with paired F344 host hearts (subject to same circulation but histologically normal), day-0 hearts, and syngrafts (P < 0.008, n = 4 at each time). Transcript levels in cardiac allografts were higher than those in paired host spleens (a major source of inflammatory cells) (P < 0.0001), indicating the localized nature of Gal/GalNAc lectin induction. By in situ hybridization and immunostaining, Gal/GalNAc lectin expression localized to a subset of inflammatory cells in cardiac allografts. These findings link Gal/GalNAc macrophage lectin to the chronic rejection process, as a possible mediator of macrophage infiltration.

[Testis yolk sec tumor--a case report].

A 16-months old normal boy, a right testicular mass was discovered incidentally by his grandmother. Right inguinal orchiectomy was performed. Histological examination showed yolk sac tumor of testis. A preoperative alpha-fetoprotein level was markedly elevated at 13568.9 ng/ml, by the end of the twelve postoperative months, the serum alpha-fetoprotein levels was 118 ng/ml. The patient is doing well one-year postoperatively.

Plasma characteristics of beef cattle classified as resistant or susceptible to horn flies.

Horn flies inflict economic losses on cattle producers. Therefore, the objective of this study was to identify a serological marker for cows that are innately resistant to horn flies. Plasma characteristics (optical density, cortisol, and protein pattern) were studied in beef cattle classified (cow-type) as horn fly resistant or susceptible. Cows (n = 30) of five breed groups were used in this study. Cows were bled via jugular venipuncture in May (the beginning of the horn fly season). Plasma optical density at wavelengths 200 and 464 nm was different (P < .05) between resistant and susceptible cattle. Cow-type also affected (P < .05) area percentage for proteins with running molecular weights (M(r)) of 74,000 and 54,000 daltons. Breed group affected (P < .05) optical density at wavelengths 200, 280, 320, and 464 nm and concentration of cortisol in plasma. When the ratio of area percentage for protein bands 7 and 9 (M(r) 74,000 and 54,000, respectively) was determined, cows could be categorized as horn fly resistant or susceptible. These data suggest that a serological marker for horn fly resistant cattle has been identified; however, the marker will need to be tested on a larger population of cattle.

Selection for arsenite resistance causes reversible changes in minicircle composition and kinetoplast organization in Leishmania mexicana.

Certain minor minicircle sequence classes in the kinetoplast DNA (kDNA) networks of arsenite- or tunicamycin-resistant Leishmania mexicana amazonensis variants whose nuclear DNA is amplified appear to be preferentially selected to replicate (S. T. Lee, C. Tarn, and K. P. Chang, Mol. Biochem. Parasitol. 58:187-204, 1993). These sequences replace the predominant wild-type minicircle sequences to become dominant species in the kDNA network. The switch from wild-type-specific to variant-specific minicircles takes place rapidly within the same network, the period of minicircle dominance changes being defined as the transition period. To investigate the structural organization of the kDNA networks during this transition period, we analyzed kDNA from whole arsenite-resistant Leishmania parasites by dot hybridization with sequence-specific DNA probes and by electron-microscopic examination of isolated kDNA networks in vitro. Both analyses concluded that during the switch of dominance the predominant wild-type minicircle class was rapidly lost and that selective replication of variant-specific minicircles subsequently filled the network step by step. There was a time during the transition when few wild-type- or variant-specific minicircles were present, leaving the network almost empty and exposing a species of thick, long, fibrous DNA which seemed to form a skeleton for the network. Both minicircles and maxicircles were found to attach to these long DNA fibrils. The nature of the long DNA fibrils is not clear, but they may be important in providing a framework for the network structure and a support for the replication of minicircles and maxicircles.