Progesterone receptor membrane component 1 is involved in oral cancer cell metastasis

Cancer metastasis is a common cause of failure in cancer therapy. However, over 60% of oral cancer patients present with advanced stage disease, and the five‐year survival rates of these patients decrease from 72.6% to 20% as the stage becomes more advanced. In order to manage oral cancer, identification of metastasis biomarker and mechanism is critical. In this study, we use a pair of oral squamous cell carcinoma lines, OC3, and invasive OC3‐I5 as a model system to examine invasive mechanism and to identify potential therapeutic targets. We used two‐dimensional differential gel electrophoresis (2D‐DIGE) and matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF/TOF MS) to examine the global protein expression changes between OC3 and invasive OC3‐I5. A proteomic study reveals that invasive properties alter the expression of 101 proteins in OC3‐I5 cells comparing to OC3 cells. Further studies have used RNA interference technique to monitor the influence of progesterone receptor membrane component 1 (PGRMC1) protein in invasion and evaluate their potency in regulating invasion and the mechanism it involved. The results demonstrated that expression of epithelial‐mesenchymal transition (EMT) markers including Twist, p‐Src, Snail1, SIP1, JAM‐A, vimentin and vinculin was increased in OC3‐I5 compared to OC3 cells, whereas E‐cadherin expression was decreased in the OC3‐I5 cells. Moreover, in mouse model, PGRMC1 is shown to affect not only migration and invasion but also metastasis in vivo. Taken together, the proteomic approach allows us to identify numerous proteins, including PGRMC1, involved in invasion mechanism. Our results provide useful diagnostic markers and therapeutic candidates for the treatment of oral cancer invasion.     

Amino acid requirement of growing pigs depending on amino acid efficiency and level of protein deposition. 2nd communication: threonine

Forty Swiss Large White pigs (barrows with 31.7 kg initial to 103.7 kg final BW) were equally and randomly assigned to one of four treatments (H0, H200, L0, L200) involving a combination of chromium supplementation (0 or 200 μg/kg) and glycemic index (high GI (H) or low GI (L)). Growth performance, energy and protein digestibility, carcass composition, and some plasma traits were investigated.

The data indicated, that the substitution of dietary carbohydrates with fat and crude fibre (low GI) resulted in lower growth performance due to impaired energy digestibility. Moreover, the strong stimulation of insulin secretion due to the high and rapid availability of carbohydrates of the diets HO and H200 caused increased carcass fat deposition. Chromium supplementation also affected plasma insulin and glucagon concentrations. Depending on glycemic index, chromium affected the growth performance. Daily gain was reduced in pigs of the L200 treatment compared to the L0 group. This finding indicated that the energy availability expressed as GI is one of several nutritional factors, which determine the efficacy of dietary chromium. We could not corroborate evidences that dietary chromium modifies the chemical composition of the whole carcass, but depending on GI, chemical composition of the longissimus muscle was affected.     

Multiple cases of asymmetric introgression among horseshoe bats detected by phylogenetic conflicts across loci

Phylogenetic discordance among taxa can provide powerful insights into past episodes of introgressive hybridization, as well as lineage sorting. Previously, we showed that the taxonomically distinct taxon Rhinolophus sinicus septentrionalis has undergone historical introgression with its sympatric sister subspecies Rhinolophus sinicus sinicus. To examine in more detail the extent of gene flow between these two taxa, and also between these and their sister species Rhinolophus thomasi, we obtained new samples from China, Myanmar, and Vietnam, and combined new and published genetic data from these, Rhinolophus rouxii, and Rhinolophus indorouxii from India. Phylogenetic analyses revealed three separate cases of discordance: between R. s. septentrionalis and adjacent populations of R. s. sinicus, between R. s. septentrionalis and R. thomasi and between eastern populations of R. s. sinicus and a newly‐identified lineage. In both former cases, the mitochondrial DNA introgression appears to be asymmetric, which is likely to have resulted from mating between R. s. septentrionalis females with smaller R. s. sinicus and R. thomasi males, although we cannot rule out other scenarios completely. Further conflicts between genetic data and accepted species arrangements across the genus, with paraphyly of members of the rouxii‐group, suggest the need for a thorough systematic revision of relationships within this group. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110 , 346–361.     

C radiolabeling of proteins to monitor biodistribution of ingested proteins

The economical preparation of microgram quantities of ¹⁴C-labeled proteins by in vacuo methylation with methyl iodide is described. The ¹⁴C radiolabeling was achieved by the covalent attachment of [¹⁴C]methyl groups onto amino and imidazole groups by reaction in vacuo with [¹⁴C]methyl iodide. The method was tested by investigating the biodistribution of ¹⁴C in rats that were fed ¹⁴C-labeled human soluble cluster of differentiation 14 (CD14) protein, a receptor for bacterial lipopolysaccharide. Two other control proteins, bovine serum albumin (BSA) and casein, were also labeled with ¹⁴C and used for comparative analysis to determine the following: (i) the efficacy and cost efficiency of the in vacuo radiolabeling procedure and (ii) the extent of incorporation of the ¹⁴C label into the organs of orogastrically fed 10-day-old Sprague–Dawley rats. [¹⁴C]BSA, [¹⁴C]casein, and [¹⁴C]CD14 were individually prepared with specific radioactivities of 34,400, 18,800, and 163,000 disintegrations per minute (dpm)/μg, respectively. It was found that the accumulation of ¹⁴C label in the organs of [¹⁴C]CD14-fed rats, most notably the persistence of ¹⁴C in the stomach 480 min postgavage, was temporally and spatially distinct from [¹⁴C]BSA and [¹⁴C]casein-fed rats.     

Effects of Hydrogen Peroxide on Wound Healing in Mice in Relation to Oxidative Damage

It has been established that low concentrations of hydrogen peroxide (H₂O₂) are produced in wounds and is required for optimal healing. Yet at the same time, there is evidence that excessive oxidative damage is correlated with poor-healing wounds. In this paper, we seek to determine whether topical application of H₂O₂ can modulate wound healing and if its effects are related to oxidative damage. Using a C57BL/6 mice excision wound model, H₂O₂ was found to enhance angiogenesis and wound closure at 10 mM but retarded wound closure at 166 mM. The delay in closure was also associated with decreased connective tissue formation, increased MMP-8 and persistent neutrophil infiltration. Wounding was found to increase oxidative lipid damage, as measured by F₂-isoprostanes, and nitrative protein damage, as measured by 3-nitrotyrosine. However H₂O₂ treatment did not significantly increase oxidative and nitrative damage even at concentrations that delay wound healing. Hence the detrimental effects of H₂O₂ may not involve oxidative damage to the target molecules studied.     

Longitudinal Liver Stiffness Assessment in Patients with Chronic Hepatitis C Undergoing Antiviral Therapy

Background/Aims

Liver stiffness (LS) measurement by means of transient elastography (TE) is accurate to predict fibrosis stage. The effect of antiviral treatment and virologic response on LS was assessed and compared with untreated patients with chronic hepatitis C (CHC).

Methods

TE was performed at baseline, and at weeks 24, 48, and 72 in 515 patients with CHC.

Results

323 treated (62.7%) and 192 untreated patients (37.3%) were assessed. LS experienced a significant decline in treated patients and remained stable in untreated patients at the end of study (P<0.0001). The decline was significant for patients with baseline LS ≥ 7.1 kPa (P<0.0001 and P 0.03, for LS ≥9.5 and ≥7.1 kPa vs lower values, respectively). Sustained virological responders and relapsers had a significant LS improvement whereas a trend was observed in nonresponders (mean percent change −16%, −10% and −2%, for SVR, RR and NR, respectively, P 0.03 for SVR vs NR). In multivariate analysis, high baseline LS (P<0.0001) and ALT levels, antiviral therapy and non-1 genotype were independent predictors of LS improvement.

Conclusions

LS decreases during and after antiviral treatment in patients with CHC. The decrease is significant in sustained responders and relapsers (particularly in those with high baseline LS) and suggests an improvement in liver damage.     

Azithromycin and Ciprofloxacin Resistance in Salmonella Bloodstream Infections in Cambodian Adults

Background

Salmonella enterica is a frequent cause of bloodstream infection (BSI) in Asia but few data are available from Cambodia. We describe Salmonella BSI isolates recovered from patients presenting at Sihanouk Hospital Centre of Hope, Phnom Penh, Cambodia (July 2007–December 2010).

Methodology

Blood was cultured as part of a microbiological prospective surveillance study. Identification of Salmonella isolates was performed by conventional methods and serotyping. Antibiotic susceptibilities were assessed using disk diffusion, MicroScan and E-test macromethod. Clonal relationships were assessed by Pulsed Field Gel Electrophoresis; PCR and sequencing for detection of mutations in Gyrase and Topoisomerase IV and presence of qnr genes.

Principal Findings

Seventy-two Salmonella isolates grew from 58 patients (mean age 34.2 years, range 8–71). Twenty isolates were identified as Salmonella Typhi, 2 as Salmonella Paratyphi A, 37 as Salmonella Choleraesuis and 13 as other non-typhoid Salmonella spp. Infection with human immunodeficiency virus (HIV) was present in 21 of 24 (87.5%) patients with S. Choleraesuis BSI. Five patients (8.7%) had at least one recurrent infection, all with S. Choleraesuis; five patients died. Overall, multi drug resistance (i.e., co-resistance to ampicillin, sulphamethoxazole-trimethoprim and chloramphenicol) was high (42/59 isolates, 71.2%). S. Typhi displayed high rates of decreased ciprofloxacin susceptibility (18/20 isolates, 90.0%), while azithromycin resistance was very common in S. Choleraesuis (17/24 isolates, 70.8%). Two S. Choleraesuis isolates were extended spectrum beta-lactamase producer.

Conclusions and Significance

Resistance rates in Salmonella spp. in Cambodia are alarming, in particular for azithromycin and ciprofloxacin. This warrants nationwide surveillance and revision of treatment guidelines.     

A Quantitative,Comparative Study of Element Variations Found within the Range of Blood Cells from the Tropical Ascidian Phallusia philippinensis,using the Nuclear Microscope

The present study examines the concentrations of vanadium, bromine and sulphur contained within cryofixed/freeze-dried blood cells of the ascidian Phallusia philippinensis. Elemental profiles of seven cell types were obtained using the National University of Singapore nuclear microscope, in order to ascertain the cell types predominantly involved in accumulation. Morula cells were found to contain the following mean values (in ppm dry weight); 7878 vanadium, 34484 bromine and 61078 sulphur. Signet ring cells contained 5191 vanadium, 23945 bromine and 15281 sulphur. Compartment cells had 606 vanadium, 20700 bromine and 24309 sulphur. Other less abundant cell types such as lymphocytes, macrogranular amoebocytes, carotenoid pigment cells and granular amoebocytes were also analysed and found to contain (in ppm) 4384, 6652, 2366 and 10246 vanadium, 19652, 15630, 5964 and 11735 bromine and 13289, 15309, 3106 and 42968 sulphur, respectively. Sulphur occurred in high levels in all cell types, which could indicate its involvement in the vanadium concentration process, while bromine, incorporated into complexes, may be utilised for anti-fouling rather than as a deterrent to predators.     

Glycation improves the thermostability of trypsin and chymotrypsin

A novel glycation procedure, in vacuo glycation, was used to attach glucose covalently to the lysine residues of trypsin and chymotrypsin. Glycated trypsin and glycated chymotrypsin have greatly increased thermostability compared to the native enzymes. For example, glycated bovine trypsin, incubated at 50 degrees C and pH 8.0 for 3 h, retained more than 50% of its original activity whereas the native enzyme was inactivated under the same conditions. Similarly, after incubation at 50 degrees C and pH 8.0, glycated bovine chymotrypsin retained 45% of its original activity and the native enzyme was inactivated. Glycated porcine trypsin is exceptionally thermostable and could be used to digest native ribonuclease at 70 degrees C without the need for prior denaturation. The apparent increase in the thermal stability of the glycated proteins observed in activity measurements is also reflected by an increase in the T(m) values determined with differential scanning calorimetry (DSC) and circular dichroism (CD). The glycation does not alter the activity or specificity of these enzymes.     

8-aminoquinolines as anticoccidials - Part III.

Analogues of the antimalarial pentaquine, 1, in which the nature of the side-chain on the 8-amino position was varied, were prepared and evaluated for anticoccidial activity both in vitro and in vivo. Specifically, both the inter-nitrogen distance and the nature of the terminal amino group were investigated. Novel analogues of equal or improved efficacy in vitro and in vivo to pentaquine were discovered.