Phenetic relationships inClerodendrum (Verbenaceae) and some phylogenetic considerations

Cluster analyses by different methods and a minimum spanning tree were used to study phenetic relationships in the genusChlerodendrum. 129 species were scored for 52 morphological characters corresponding to 119 character states. The phenetic results suggest a classification into 7 distinct groups, which may be grouped into two subgenera. This classification is supported by the iridoid distribution as well as by some phylogenetic considerations.     

The use of the rat hepatoma cell line MH1C1 to investigate the stimulus-secretion response of hepatic acute phase proteins

The rat hepatoma cell line MH1C1 has been characterized to show a stimulated secretion of C-reactive protein in response to both leukocyte supernatant and a purified human interleukin-1 preparation. The time-dependency and dose-response relationship of CRP secretion were comparable to and somewhat more sensitive than the effects of leukocyte supernatant and purified human interleukin-1 on the proliferative rate of murine thymocytes; the proliferative rate of the hepatoma cell line MH1C1 was unchanged under these conditions. Agents which affect the thymocyte bioassay response to interleukin-1 namely interleukin-2, lipopolysaccharide, concanavalin A and phytohemagglutinin showed no effect on the C-reactive protein release of the MH1C1 cell line. These data strongly support the suitability of this cell line for the in vitro study of the hepatic acute phase stimulus-secretion response.     

Conjunctival melanoma copy number alterations and correlation with mutation status,tumor features,and clinical outcome

Relatively little is known about the genetic aberrations of conjunctival melanomas (CoM) and their correlation with clinical and histomorphological features as well as prognosis. The aim of this large collaborative multicenter study was to determine potential key biomarkers for metastatic risk and any druggable targets for high metastatic risk CoM. Using Affymetrix single nucleotide polymorphism genotyping arrays on 59 CoM, we detected frequent amplifications on chromosome (chr) 6p and deletions on 7q, and characterized mutation‐specific copy number alterations. Deletions on chr 10q11.21‐26.2, a region harboring the tumor suppressor genes, PDCD4, SUFU, NEURL1, PTEN, RASSF4, DMBT1, and C10orf90 and C10orf99, significantly correlated with metastasis (Fisher's exact, p ≤ 0.04), lymphatic invasion (Fisher's exact, p ≤ 0.02), increasing tumor thickness (Mann–Whitney, p ≤ 0.02), and BRAF mutation (Fisher's exact, p ≤ 0.05). This enhanced insight into CoM biology is a step toward identifying patients at risk of metastasis and potential therapeutic targets for systemic disease.     

The Brain Response to Peripheral Insulin Declines with Age: A Contribution of the Blood-Brain Barrier?

ObjectivesIt is a matter of debate whether impaired insulin action originates from a defect at the neural level or impaired transport of the hormone into the brain. In this study, we aimed to investigate the effect of aging on insulin concentrations in the periphery and the central nervous system as well as its impact on insulin-dependent brain activity.MethodsInsulin, glucose and albumin concentrations were determined in 160 paired human serum and cerebrospinal fluid (CSF) samples. Additionally, insulin was applied in young and aged mice by subcutaneous injection or intracerebroventricularly to circumvent the blood-brain barrier. Insulin action and cortical activity were assessed by Western blotting and electrocorticography radiotelemetric measurements.ResultsIn humans, CSF glucose and insulin concentrations were tightly correlated with the respective serum/plasma concentrations. The CSF/serum ratio for insulin was reduced in older subjects while the CSF/serum ratio for albumin increased with age like for most other proteins. Western blot analysis in murine whole brain lysates revealed impaired phosphorylation of AKT (P-AKT) in aged mice following peripheral insulin stimulation whereas P-AKT was comparable to levels in young mice after intracerebroventricular insulin application. As readout for insulin action in the brain, insulin-mediated cortical brain activity instantly increased in young mice subcutaneously injected with insulin but was significantly reduced and delayed in aged mice during the treatment period. When insulin was applied intracerebroventricularly into aged animals, brain activity was readily improved.ConclusionsThis study discloses age-dependent changes in insulin CSF/serum ratios in humans. In the elderly, cerebral insulin resistance might be partially attributed to an impaired transport of insulin into the central nervous system.     

Fabrication of hierarchical hybrid structures using bio-enabled layer-by-layer self-assembly

Development of versatile and flexible assembly systems for fabrication of functional hybrid nanomaterials with well-defined hierarchical and spatial organization is of a significant importance in practical nanobiotechnology applications. Here we demonstrate a bio-enabled self-assembly technique for fabrication of multi-layered protein and nanometallic assemblies utilizing a modular gold-binding (AuBP1) fusion tag. To accomplish the bottom-up assembly we first genetically fused the AuBP1 peptide sequence to the C'-terminus of maltose-binding protein (MBP) using two different linkers to produce MBP-AuBP1 hetero-functional constructs. Using various spectroscopic techniques, surface plasmon resonance (SPR) and localized surface plasmon resonance (LSPR), we verified the exceptional binding and self-assembly characteristics of AuBP1 peptide. The AuBP1 peptide tag can direct the organization of recombinant MBP protein on various gold surfaces through an efficient control of the organic-inorganic interface at the molecular level. Furthermore using a combination of soft-lithography, self-assembly techniques and advanced AuBP1 peptide tag technology, we produced spatially and hierarchically controlled protein multi-layered assemblies on gold nanoparticle arrays with high molecular packing density and pattering efficiency in simple, reproducible steps. This model system offers layer-by-layer assembly capability based on specific AuBP1 peptide tag and constitutes novel biological routes for biofabrication of various protein arrays, plasmon-active nanometallic assemblies and devices with controlled organization, packing density and architecture.     

Biological responses in Balb/c mice after long‐term parenteral administration of the light subunit of mushroom tyrosinase

Light subunit of mushroom tyrosinase (LSMT) is a protein of unknown function from mushroom Agaricus bisporus that has been demonstrated to permeate through rat intestine ex vivo. Thus, it can be absorbed in the intestine, thereby holding a promise as a drug carrier for oral administration, similar to HA‐33 protein from botulinum, one of the closest structural homologs of LSMT. However, the safety of LSMT should be ensured prior to its use. Here, we described biological response of LSMT upon weekly intraperitoneal administration of 50 μg/day to the Balb/c mice for 12 weeks. Motoric and behavior profiles, as well as the index of main organs (liver, spleen, lung, heart, and kidney), and body weight, were not significantly changed as compared with the control group. Also, no IgG was detected in the serum. The results suggest that LSMT is safe for further development.     

Intermedilysin induces EGR-1 expression through calcineurin/NFAT pathway in human cholangiocellular carcinoma cells

RNF8 is a nuclear protein having an N-terminal forkhead-associated (FHA) domain and a C-terminal RING-finger (RF) domain. Depletion of RNF8 caused cell growth inhibition and cell cycle arrest at not only S but also G2/M phases. In addition, cell death was frequently observed in RNF8-depleted cells. Analyses of time-lapse microscopy revealed that the cells died in mitosis and interphase. To elucidate the RNF8 function in M phase, the Plk1 content in RNF8-depleted cells was examined. The amount of RNF8 decreased time-dependently, whereas Plk1 reciprocally increased by transfection of RNF8 siRNA. Protein contents of RNF8 and Plk1 among various cell lines were also compared. RNF8 in normal cell lines was much higher than that in many cancer cell lines. Conversely, Plk1 in normal cell lines was lower than in cancer cell lines. These results suggest that RNF8 is downregulated in many cancer cells and inversely correlated with Plk1.     

Glucose-Raising Polymorphisms in the Human Clock Gene Cryptochrome 2 (CRY2) Affect Hepatic Lipid Content

Circadian rhythms govern vital functions. Their disruption provokes metabolic imbalance favouring obesity and type-2 diabetes. The aim of the study was to assess the role of clock genes in human prediabetes. To this end, genotype-phenotype associations of 121 common single nucleotide polymorphisms (SNPs) tagging ARNTL, ARNTL2, CLOCK, CRY1, CRY2, PER1, PER2, PER3, and TIMELESS were assessed in a study population of 1,715 non-diabetic individuals metabolically phenotyped by 5-point oral glucose tolerance tests. In subgroups, hyperinsulinaemic-euglycaemic clamps, intravenous glucose tolerance tests, and magnetic resonance imaging/spectroscopy were performed. None of the tested SNPs was associated with body fat content, insulin sensitivity, or insulin secretion. Four CRY2 SNPs were associated with fasting glycaemia, as reported earlier. Importantly, carriers of these SNPs’ minor alleles revealed elevated fasting glycaemia and, concomitantly, reduced liver fat content. In human liver tissue samples, CRY2 mRNA expression was directly associated with hepatic triglyceride content. Our data may point to CRY2 as a novel switch in hepatic fuel metabolism promoting triglyceride storage and, concomitantly, limiting glucose production. The anti-steatotic effects of the glucose-raising CRY2 alleles may explain why these alleles do not increase type-2 diabetes risk.     

Diet of Chelidonichthys obscurus and Chelidonichthys lastoviza (Pisces: Triglidae) from the Gulf of Gabes (Tunisia)

The most common gurnard species in the Gulf of Gabes (Tunisia) are Chelidonichthys lastoviza and C. obscurus. These two species were collected by trawl nets between February 2000 and July 2002 at depths ranging from 80 to 150 meters. In the laboratory the stomach contents were examined for diet assessment and to determine diet variation in relation to fish size, season and sex. The importance of different prey items was analysed utilising several feeding indexes, while diet overlap was tested using Schoener's index and the null model test. The two species showed similar diets, with the same qualitative and quantitative composition. Basic food consisted of crustaceans, mostly mysids and decapods. For Chelidonichthys lastoviza:Gastrosaccus sp. (F = 48.57%), Anchialina agilis (F = 30.57%) and Sicyonia carinata (F = 21.71%); for Chelidonichthys obscurus:Gastrosaccus sp. (F = 80.95%), A. agilis (F = 55.41%) and Pontocaris lacazei (F = 15.15%). A change in food composition based on fish size was observed in both species: mysids more frequently in the diet of small specimens, and decapods in the largest fishes. Seasonal variation in their diet was also recorded. The two fishes showed a significantly higher number of empty stomachs in late autumn and winter. Regardless of species and size class, a biologically significant diet overlap was observed between C. lastoviza and C. obscurus. This result may indicate a lack of competition among triglid species size‐class combinations or strong competition that has not yet led to resource portioning.     

Furnished cages for laying hens: study of the effects of group size and litter provision on laying location, zootechnical performance and egg quality

The furnished cage is a new housing system for layers. A current trend in furnished cage design is to increase group size and replace the litter box with a mat provided with litter. An experiment was set up to determine the effects of group size and litter provision on laying performance and egg quality of beak-trimmed ISA Brown hens housed in large furnished cages with more than 12 hens. Six treatments, each of 18 furnished cages (768 cm(2)/hen including nest and litter area) were compared in a 3 × 2 experimental trial: three group sizes (S20 (20 hens per cage), S40 (40) and S60 (60)), with or without feed as litter distributed on the mat of the litter area. The provision of facilities per hen was equal in all treatments. Mortality, laying rate, mean egg weight, feed intake and feed conversion ratio were unaffected by group size over the 53-week laying period, and performance exceeded the ISA production standards. The overall percentage of eggs laid in the nest exceeded 95% except that it was slightly lower in group S20 (92.0% ± 6.4% v. S40: 96.0% ± 3.3% and S60: 96.2% ± 2.7%) leading to a higher proportion of dirty eggs (S20: 1.6% ± 2.2%, S40: 1.4% ± 1.5%, S60: 1.0% ± 1.0%). At 66 to 70 weeks, eggs laid outside the nest had a slightly higher count of mesophilic bacteria on the eggshell (5.0 log CFU/egg ± 0.4) than those laid in the nest (4.8 log CFU/egg ± 0.5) but no difference in contamination was observed between group sizes. Litter provision had no effect on mortality, egg weight or egg quality traits except for a higher proportion of broken eggs in cages with litter (5.3% ± 6.2% v. 4.6% ± 5.7%). Providing hens with feed for litter was associated with a higher laying rate (97.3% ± 3.2% v. 94.8% ± 4.4% at 23 weeks) and an apparent improvement in feed efficiency at the beginning of the laying period (feed conversion ratio based on feed consumption at the trough: 2.18 ± 0.06 with litter v. 2.28 ± 0.09 without litter at 25 weeks). The results of this study showed that a high level of productivity and good egg quality could be obtained in large furnished cages. Further research is needed to assess the impact on hens' welfare and performance of using more economically competitive substrates than feed for litter.