Synthesis and stability in biological media of 1H-imidazole-1-carboxylates of ROS203, an antagonist of the histamine H3 receptor

A series of carbamate derivatives of the H(3) antagonist ROS203 (1) were prepared, and their lipophilicity and steric hindrance were modulated by introducing linear or branched alkyl chains of various lengths. In vitro stability studies were conducted to evaluate how structural modulations affect the intrinsic reactivity of the carbamoyl moiety and its recognition by metabolic enzymes. Linear alkyl carbamates were the most susceptible to enzymatic hydrolysis, with bioconversion rates being higher in rat liver and plasma. Chain ramification significantly enhanced the enzymatic stability of the set, with two derivatives (1g and 1h) being more stable by a factor of 8-40 than the ethyl carbamate 1a. Incubation with bovine serum albumin (BSA) showed a protective role of proteins on chemical and porcine-liver esterase (PLE)-catalyzed hydrolysis. Ex vivo binding data after i.v. administration of 1h revealed prolonged displacement of the labeled ligand [(3)H]-(R)-alpha-methylhistamine ([(3)H]RAMHA) from rat-brain cortical membranes, when compared to 1. However, the high rates of bioconversion in liver, as well as the chemical instability of 1h, suggest that further work is needed to optimize the enzymatic and chemical stability of these compounds.     

The CAPN10 gene is associated with insulin resistance phenotypes in the Spanish population

Cardiovascular disease is the leading cause of morbidity and mortality in the industrialized world. Familial aggregation of cardiovascular risk factors is a frequent finding, but genetic factors affecting its presentation are still poorly understood. The calpain 10 gene (CAPN10) has been associated with type 2 diabetes (T2DM), a complex metabolic disorder with increased risk of cardiovascular disease. Moreover, the CAPN10 gene has been associated with the presence of metabolic syndrome (MS) in T2DM and in polycystic ovary syndrome (PCOS). In this work, we have analysed whether the polymorphisms UCSNP44, -43, -19 and -63 are related to several cardiovascular risk factors in the context of MS. Molecular analysis of CAPN10 gene was performed in 899 individuals randomly chosen from a cross-sectional population-based epidemiological survey. We have found that CAPN10 gene in our population is mainly associated with two indicators of the presence of insulin resistance: glucose levels two hours after a 75-g oral glucose tolerance test (OGTT) and HOMA values, although cholesterol levels and blood pressure values are also influenced by CAPN10 variants. In addition, the 1221/1121 haplogenotype is under-represented in individuals that fulfil the International Diabetes Federation (IDF) diagnostic criteria for MS. Our results suggest that CAPN10 gene is associated with insulin resistance phenotypes in the Spanish population.     

The synergistic effect of concomitant schistosomiasis, hookworm, and trichuris infections on children's anemia burden

Objective

To estimate the degree of synergism between helminth species in their combined effects on anemia.

Methods

Quantitative egg counts using the Kato–Katz method were determined for Ascaris lumbricoides, hookworm, Trichuris trichiura, and Schistosoma japonicum in 507 school-age children from helminth-endemic villages in The Philippines. Infection intensity was defined in three categories: uninfected, low, or moderate/high (M+). Anemia was defined as hemoglobin <11 g/dL. Logistic regression models were used to estimate odds ratios (OR), 95% confidence intervals (CI), and synergy index for pairs of concurrent infections.

Results

M+ co-infection of hookworm and S. japonicum (OR = 13.2, 95% CI: 3.82–45.5) and of hookworm and T. trichiura (OR = 5.34, 95% CI: 1.76–16.2) were associated with higher odds of anemia relative to children without respective M+ co-infections. For co-infections of hookworm and S. japonicum and of T. trichiura and hookworm, the estimated indices of synergy were 2.9 (95% CI: 1.1–4.6) and 1.4 (95% CI: 0.9–2.0), respectively.

Conclusion

Co-infections of hookworm and either S. japonicum or T. trichiura were associated with higher levels of anemia than would be expected if the effects of these species had only independent effects on anemia. This suggests that integrated anti-helminthic treatment programs with simultaneous deworming for S. japonicum and some geohelminths could yield a greater than additive benefit for reducing anemia in helminth-endemic regions.     

Alternate AChE-R variants facilitate cellular metabolic activity and resistance to genotoxic stress through enolase and RACK1 interactions

Tumorogenic transformation is a multifaceted cellular process involving combinatorial protein-protein interactions that modulate different cellular functions. Here, we report apparent involvement in two independent tumorogenic processes by distinct partner protein interactions of the stress-induced acetylcholinesterase AChE-R and N-AChE-R variants. Human testicular tumors showed elevated levels of N-terminally extended N-AChE-R compared with healthy tissue, indicating alternate promoter usage in the transformed cells. Two-hybrid screens demonstrate that the C-terminus common to both N-AChE-R and AChE-R interacts either with the glycolytic enzyme enolase or with the scaffold protein RACK1. In vitro, the AChE-R C-terminal peptide ARP elevated enolase's activity by 12%, suggesting physiological relevance for this interaction. Correspondingly, CHO cells expressing either human AChE-R or N-AChE-R but not AChE-S showed a 25% increase in cellular ATP levels, indicating metabolic significance for this upregulation of enolase activity. ATP levels could be reduced by AChE-targeted siRNA in CHO cells expressing AChE-R but not AChE-S, attributing this elevation to the AChE-R C-terminus. Additionally, transfected CHO cells expressing AChE-R but not N-AChE-R showed resistance to up to 60muM of the common chemotherapeutic agent, cis-platinum, indicating AChE-R involvement in another molecular pathway. cis-Platinum elevates the expression of the apoptosis-regulator p53-like protein, p73, which is inactivated by interaction with the scaffold protein RACK1. In co-transfected cells, AChE-R competed with endogenous RACK1 for p73 interaction. Moreover, AChE-R-transfected CHO cells presented higher levels than control cells of the pro-apoptotic TAp73 as well as the anti-apoptotic dominant negative DeltaNp73 protein, leading to an overall decrease in the proportion of pro-apoptotic p73. Together, these findings are compatible with the hypothesis that in cancer cells, both AChE-R and N-AChE-R elevate cellular ATP levels and that AChE-R modifies p73 gene expression by facilitating two independent cellular pathways, thus conferring both a selective metabolic advantage and a genotoxic resistance.     

Signal molecules in the peanut–bradyrhizobia interaction

Main nodulation signal molecules in the peanut–bradyrhizobia interaction were examined. Flavonoids exuded by Arachis hypogaea L. cultivar Tegua were genistein, daidzein and chrysin, the latest being released in lower quantities. Thin layer chromatography analysis from genistein-induced bacterial cultures of three peanut bradyrhizobia resulted in an identical Nod factor pattern, suggesting low variability in genes involved in the synthesis of these molecules. Structural study of Nod factor by mass spectrometry and NMR analysis revealed that it shares a variety of substituents with the broad-host-range Rhizobium sp. NGR234 and Bradyrhizobium spp. Nodulation assays in legumes nodulated by these rhizobia demonstrated differences between them and the three peanut bradyrhizobia. The three isolates were classified as Bradyrhizobium sp. Their fixation gene nifD and the common nodulation genes nodD and nodA were also analyzed. Accession numbers: AY427207, EF202193, EF158295 (16S rRNA gene of strains NLH25, NOD31 and NDEHE, respectively); DQ295199, DQ295200, DQ295201 (Partial nifD gene sequences of strains NLH25, NOD31 and NDEHE, respectively).     

Reciprocal changes in calcification of the gastrolith and cuticle during the molt cycle of the red claw crayfish Cherax quadricarinatus

Mobilization of calcium during the molt cycle from the cuticle to transient calcium deposits is widely spread in crustaceans. The dynamics of calcium transport to transient calcium deposits called gastroliths and to the cuticle over the course of the molt cycle were studied in the crayfish Cherax quadricarinatus. In this species, calcium was deposited in the gastroliths during premolt and transported back to the cuticle during postmolt, shown by digital X-ray radiograph analysis. The predominant mineral in the crayfish is amorphous calcium carbonate embedded in an organic matrix composed mainly of chitin. Scanning electron micrographs of the cuticle during premolt showed that the endocuticle and parts of the exocuticle were the source of most of the labile calcium, while the epicuticle did not undergo degradation and remained mineralized throughout the molt cycle. The gastroliths are made of concentric layers of amorphous calcium carbonate intercalated between chitinous lamella. Measurements of pH and calcium levels during gastrolith deposition showed that calcium concentrations in the gastroliths, stomach, and muscle were about the same (10 to 11 mmol l(-1)). On the other hand, pH varied greatly, from 8.7+/-0.15 in the gastrolith cavity through 7.6+/-0.2 in muscle to 6.9+/-0.5 in the stomach.     

Serum bilirubin constituents in different experimental models of conjugated hyperbilirubinemia

The object of the study was to explain the differences in bilirubin level in various experimental jaundice models. The Bil constituents in conjugated hyperbilirubinemic dog models were identified. Total Bil was measured using the Jendrassik and Grof method and direct spectrophotometry. Conjugated and unconjugated Bil were measured using the Weber-Schalm extraction method. Bilirubin covalently bound to albumin was measured indirectly from the total Bil and the non-CBBA fraction. The non-CBBA was estimated either as the sum of the CB and UCB concentrations determined by the W-S method or as the nonprecipitated fraction after deproteinization with ammonium sulfate-saturated ethanol when using DS. The TB, CB, UCB, and CBBA levels were compared in two hyperbilirubinemic dog models: chronic bile duct ligation (CBDL) and internal choledochocaval anastomosis (CDCA). The mean TB in internal CDCA (16.5 +/- 3.67 mg%) was significantly higher than in CBDL (3.4 +/- 1.75 mg%). Most of the serum Bil in these two models was conjugated, 13.4 +/- 2.24 and 3.2 +/- 1.7 mg%, respectively. No CBBA was found in the CBDL or in the partially obstructed internal-CDCA dogs. The TB in an external-CDCA model was essentially similar to the internal-CDCA model. The indirect Bil level in the external-CDCA model was six to seven times higher than the UCB level, and the CBBA level varied between 30 and 80% of the TB. Up to 50% CBBA was found also in patients with intra- or extrahepatic cholestasis. The findings indicate that, unlike the commonly assumed hypothesis, the serum TB level in the CDCA models, which was higher than in the CBCL one, is not due to high UCB levels. Rather, hyperbilirubinemia in the external-CDCA model is due to increased levels of CBBA or, perhaps, to variations in the amount and/or composition of CB entering the blood and cleared by the kidney.     

Virucidal Nanofiber Textiles Based on Photosensitized Production of Singlet Oxygen

Novel biomaterials based on hydrophilic polycaprolactone and polyurethane (Tecophilic®) nanofibers with an encapsulated 5,10,5,20-tetraphenylporphyrin photosensitizer were prepared by electrospinning. The doped nanofiber textiles efficiently photo-generate O₂(¹Δ_g), which oxidize external chemical and biological substrates/targets. Strong photo-virucidal effects toward non-enveloped polyomaviruses and enveloped baculoviruses were observed on the surface of these textiles. The photo-virucidal effect was confirmed by a decrease in virus infectivity. In contrast, no virucidal effect was detected in the absence of light and/or the encapsulated photosensitizer.     

Cluster of three cases of invasive meningococcal disease in a preschool facility in West Bohemia, the Czech Republic

From 1993 to 2009, there was only one cluster of invasive meningococcal disease (IMD) reported in a community of children in the Czech Republic. This exceptional cluster that occurred in a preschool facility is the focus of this report. In response to the announcement of the disease, anti-epidemic precautions were put in place. Neisseria meningitidis isolates were delivered from local laboratories to the National Reference Laboratory for Meningococcal Infections in Prague. Phenotyping was performed there along with multilocus sequence typing. Related factors and microbiological results were analyzed retrospectively. In October 2009, three girls contracted IMD within a period of 1 week in a 42-member group in a preschool facility attached to the elementary school in Starý Plzenec-Sedlec. In relation to three cases of the disease, another 66 people were registered of which 58 underwent a microbiological examination. N. meningitidis was detected in a total of five (8.6 %) people. The National Reference Laboratory for Meningococcal Infections defined the type of the strain to be C: P1.18-1,34-2,38: F1-7: ST-467 (cc269) and penA27. Tests showed the precise identity of all strains obtained from the three sick children and of two strains contracted through contact with the preschool facility. Despite the complete recovery of all patients with no permanent damage, the need for rapid cooperation between clinical sites, diagnostic laboratories, and epidemiologists was confirmed.     

Organ/tissue-specific changes in the mitochondrial genome organization of in-vitro cultures derived from different explants of a single wheat variety

Summary We have previously shown that the mitochondrial genome of long-term tissue cultures prepared from immature embryos of several varieties of cultivated wheat underwent variety-specific rearrangements resulting from either changes in the relative amounts of subgenomic components or from the appearance of novel genomic configurations. In the present work, both categories of rearrangements were studied in long-term tissue cultures initiated from other explants (shoot meristem, young leaf base, young root tip, immature inflorescence) of the same wheat variety (Chinese Spring) and were compared to those previously obtained with immature embryo cultures. Two main patterns of reorganization were found in a region of the mitochondrial genome known to be hypervariable in structure. In addition, some of the novel subgenomic configurations were obviously organ/tissue-specific whereas others were present in more than one type of organ. In several instances, the age of culture was found to determine the degree of mitochondrial DNA rearrangement. The data presented in this study strengthen the hypothesis of an association between a particular organization of the mitochondrial genome in tissue culture and its regeneration capacity.