Dynamic change and derivation process of FC and DFC through G1, S and G2 phases in HeLa cells

In order to get a deeper understanding of the relationship between nucleolus structure and its function, the dynamic change and derivation of FC (fibrillar center) and DFC (dense fibrillar component) through interphase were investigated in HeLa cells synchronized at the ultrastructural level. The results showed that there was a process of FC and DFC derivation in the nucleolus of HeLa cells during interphase. In G1 phase there were a few big FCs in the nucleolus of the HeLa cell. In S phase DFC around the FC got thickened and the configuration of the DFC changed. A lot of tiny FCs were derived from parts of the thickened DFC. We called the FC and DFC formed in G1 phase as primary FC (pri-FC) and primary DFC (pri-DFC) and the FC and DFC derived from the thickened pri-DFC as secondary FC (sec-FC) and secondary DFC (sec-DFC). In G2 phase sec-FC and sec-DFC were gradually separated from pri-DFC and scattered evenly in the nucleolus. Few large pri-FCs coexisted with numerous tiny sec-FCs in the nucleolus of HeLa cells in G2 phase. Based on the results of our observation, we suggest here a model of the dynamic change and the process of derivation of FC and DFC through interphase.     

Epac1‐induced cellular proliferation in prostate cancer cells is mediated by B‐Raf/ERK and mTOR signaling cascades

cAMP‐dependent, PKA‐independent effects on cell proliferation are mediated by cAMP binding to EPAC and activation of Rap signaling. In this report, we employed the analogue 8‐CPT‐2‐O‐Me‐cAMP to study binding to EPAC and subsequent activation of B‐Raf/ERK and mTOR signaling in human cancer cells. This compound significantly stimulated DNA synthesis, protein synthesis, and cellular proliferation of human 1‐LN prostate cancer cells. By study of phosphorylation‐dependent activation, we demonstrate that EPAC‐mediated cellular effects require activation of the B‐Raf/ERK and mTOR signaling cascades. RNAi directed against EPAC gene expression as well as inhibitors of ERK, PI 3‐kinase, and mTOR were employed to further demonstrate the role of these pathways in regulating prostate cancer cell proliferation. These studies were then extended to several other human prostate cancer cell lines and melanoma cells with comparable results. We conclude that B‐Raf/ERK and mTOR signaling play an essential role in cAMP‐dependent, but PKA‐independent, proliferation of cancer cells. J. Cell. Biochem. 108: 998–1011, 2009. © 2009 Wiley‐Liss, Inc.     

Ecology of the gastrointestinal parasites of Colobus vellerosus at Boabeng‐Fiema,Ghana: Possible anthropozoonotic transmission

Parasite richness and prevalence in wild animals can be used as indicators of population and ecosystem health. In this study, the gastrointestinal parasites of ursine colobus monkeys (Colobus vellerosus) at the Boabeng‐Fiema Monkey Sanctuary (BFMS), Ghana, were investigated. BFMS is a sacred grove where monkeys and humans have long lived in relatively peaceful proximity. Fecal samples (n = 109) were collected opportunistically from >27 adult and subadult males in six bisexual groups and one all‐male band from July 2004 to August 2005. Using fecal floatation, we detected three protozoans (two Entamoeba sp., Isospora sp.), five nematodes (Ascaris sp., Enterobius sp., Trichuris sp., two strongyle sp.), and one digenean trematode. Using fluorescein labeled antibodies, we detected an additional protozoan (Giardia sp.), and with PCR techniques, we characterized this as G. duodenalis Assemblage B and also identified a protistan (Blastocystis sp., subtype 2). The most prevalent parasite species were G. duodenalis and Trichuris sp. Parasites were more prevalent in the long wet season than the long dry. Parasite prevalence did not vary by age, and average parasite richness did not differ by rank for males whose status remained unchanged. However, males that changed rank tended to show higher average parasite richness when they were lower ranked. Individuals that spent more time near human settlements had a higher prevalence of Isospora sp. that morphologically resembled the human species I. belli. The presence of this parasite and G. duodenalis Assemblage B indicates possible anthropozoonotic and/or zoonotic transmission between humans and colobus monkeys at this site. Am J Phys Anthropol, 2009. © 2009 Wiley‐Liss, Inc.     

Resilience of inshore, juvenile snapper Pagrus auratus to angling and release

This study assessed the mortality of 157 snapper Pagrus auratus (9-29 cm, total length, L(T) ) after being conventionally angled and then released into cages (along with 48 controls) for 4 days off south-eastern Australia. Fatalities were restricted to 12 angled fish (7·6%) and mostly attributed to the ingestion of hooks and especially their subsequent removal, which caused substantial blood loss and immediate death. Hook ingestion was significantly biased towards smaller fish (<21 cm L(T)) and attributed to a lower chance of anglers initially detecting these individuals on the line (allowing them to consume more of the baits). While mortalities might be reduced in future via (1) choosing terminal rigs that promote mouth hooking and (2) cutting the line on any-hook ingested fish, the results nevertheless validate releasing unwanted angled inshore juvenile P. auratus as a means for managing their exploitation.     

Long term recovery of median nerve repair using laser‐activated chitosan adhesive films

Sutures remain the standard peripheral nerve repair technique, whether applied directly or indirectly to nerve tissue. Unfortunately, significant postoperative complications can result, such as inflammation, neuroma formation and foreign body reactions. Photochemical‐tissue‐bonding (PTB) using rose Bengal (RB) integrated into a chitosan bioadhesive is an alternative nerve repair device that removes the need for sutures. Rats were arranged into three groups: RB‐chitosan adhesives‐repair, end‐to‐end epineural suture‐repair (surgical standard) and sham laser‐irradiated control. Groups were compared through histological assessment, electrophysiological recordings and grip motor strength. RB‐chitosan adhesive repaired nerves displayed comparable results when compared to the standard suture‐repair based on histological and electrophysiological findings. Functionally, RB‐chitosan adhesive was associated with a quicker and more pronounced recovery of grip force when compared to the suture‐repair. (© 2013 WILEY‐VCH Verlag GmbH &Co. KGaA, Weinheim)     

Prophylactic role of D-Saccharic acid-1,4-lactone in tertiary butyl hydroperoxide induced cytotoxicity and cell death of murine hepatocytes via mitochondria-dependent pathways

D-Saccharic acid 1,4-lactone (DSL) is a derivative of D-glucaric acid. It is a beta-glucuronidase inhibitor and possesses anticarcinogenic, detoxifying, and antioxidant properties. In the present study, the protective effects of DSL were investigated against tertiary butyl hydroperoxide (TBHP) induced cytotoxicity and cell death in vitro using murine hepatocytes. Exposure of TBHP caused a reduction in cell viability, enhanced the membrane leakage, and disturbed the intracellular antioxidant machineries in murine hepatocytes. Investigating the signaling mechanism of TBHP-induced cellular pathophysiology and protective action of DSL, we found that TBHP exposure disrupted mitochondrial membrane potential, facilitated cytochrome c release in the cytosol, and led to apoptotic cell death via mitochondria-dependent pathways. DSL counteracted these changes and maintained normalcy in hepatocytes. Combining, results suggest that DSL possesses the ability to ameliorate TBHP-induced oxidative insult, cytotoxicity, and apoptotic cell death probably due to its antioxidant activity and functioning via mitochondria-dependent pathways.     

Evaluation and recommendations for greater accessibility of colour figures in ornithology

People who are colour‐blind or have some form of colour vision deficiency form an invisible minority and scientists should strive to be as inclusive as possible. We reviewed 2873 figures published in 2019 from 1031 scientific papers in 27 ornithological journals to determine those that were colour‐blind compatible, and those that were black‐and‐white printer friendly. About 26% of the published figures were in colour, and while most were colour‐blind compatible, only ~ 60% of them were black‐and‐white printer friendly. Ensuring figures in all forms of scientific communication can be interpreted by readers who are colour‐blind, and can be printed in black‐and‐white will improve the accessibility of ornithological research.     

Mechanism and rate constants of the Cdc42 GTPase binding with intrinsically disordered effectors

Intrinsically disordered proteins (IDPs) are often involved in signaling and regulatory functions, through binding to cellular targets. Many IDPs undergo disorder‐to‐order transitions upon binding. Both the binding mechanisms and the magnitudes of the binding rate constants can have functional importance. Previously we have found that the coupled binding and folding of any IDP generally follows a sequential mechanism that we term dock‐and‐coalesce, whereby one segment of the IDP first docks to its subsite on the target surface and the remaining segments subsequently coalesce around their respective subsites. Here we applied our TransComp method within the framework of the dock‐and‐coalesce mechanism to dissect the binding kinetics of two Rho‐family GTPases, Cdc42 and TC10, with two intrinsically disordered effectors, WASP and Pak1. TransComp calculations identified the basic regions preceding the GTPase binding domains (GBDs) of the effectors as the docking segment. For Cdc42 binding with both WASP and Pak1, the calculated docking rate constants are close to the observed overall binding rate constants, suggesting that basic‐region docking is the rate‐limiting step and subsequent conformational coalescence of the GBDs on the Cdc42 surface is fast. The possibility that conformational coalescence of the WASP GBD on the TC10 surface is slow warrants further experimental investigation. The account for the differences in binding rate constants among the three GTPase‐effector systems and mutational effects therein yields deep physical and mechanistic insight into the binding processes. Our approach may guide the selection of mutations that lead to redesigned binding pathways. Proteins 2016; 84:674–685. © 2016 Wiley Periodicals, Inc.     

Estimation of the Parameters of two Parallel Regression Lines Under Uncertain Prior Information

The problem of parallelism for bi‐linear regression lines arises in many real life investigations. For two linear regression models with normal errors, the estimation of the slope as well as the intercept parameters is considered when it is apriori suspected that the two lines are parallel. Three different estimators are defined by using both the sample data and the non‐sample uncertain prior information. The relative performances of the unrestricted, restricted and preliminary test estimators are investigated based on the analysis of the bias, and risk functions under quadratic loss. An example based on a medical study is used to illustrate the method.     

Blood lactate loads of redthroat emperor Lethrinus miniatus associated with angling stress and exhaustive exercise

Baseline, post‐angling and maximum attainable blood lactate concentrations were measured for the fishery species redthroat emperor Lethrinus miniatus to gain insight into the condition of fish released following c. 30 s angling and <45 s air exposure. Mean ± s.d . baseline blood lactate was 1·5 ± 0·6 mmol l^?1, which increased and plateaued around 6 mmol l^?1 at 15–30 min post‐angling. These values were significantly lower than those obtained from fish maximally exhausted with a prolonged chase and air exposure protocol following capture (10·9 ± 1·8 mmol l^?1), suggesting that L. miniatus is not maximally exhausted during standard angling practices.     

Multiscale Morphological and Electrical Characterization of Charge Transport Limitations to the Power Performance of Positive Electrode Blends for Lithium‐Ion Batteries

In this work, exhaustive characterizations of 3D geometries of LiNi_1/3Mn_1/3Co_1/3O₂ (NMC), LiFePO₄ (LFP), and NMC/LFP blended electrodes are undertaken for rational interpretation of their measured electrical properties and electrochemical performance. X‐ray tomography and focused ion beam in combination with scanning electron microscopy tomography are used for a multiscale analysis of electrodes 3D geometries. Their multiscale electrical properties are measured by using broadband dielectric spectroscopy. Finally, discharge rate performance are measured and analyzed by simple, yet efficient methods. It allows us to discriminate between electronic and ionic wirings as the performance limiting factors, depending on the discharge rate. This approach is a unique exhaustive analysis of the experimental relationships between the electrochemical behavior, the transport properties within the electrode, and its 3D geometry.     

Testing the suitability of insect orders as indicators for olive farming systems

• 1 A previous study suggested the use of certain insects groups as indicators for detecting organic olive farming in Southern Spain. To validate the use of these groups, insects were collected from olive orchards in Cordoba and Granada, comprising two Andalusian provinces with different surrounding landscapes.
• 2 Canopies were sampled using the branch‐beating technique during pre‐blooming and post‐blooming periods over 3 years in Granada (1999, 2000 and 2003) and 1 year in Cordoba (2003).
• 3 Using a nonparametric linear discriminant analysis method, based on the k‐nearest neighbour algorithm, two discriminant functions were constructed. A first discriminant model took into account interannual variability in Granada Province and the second model focused on environmental heterogeneity between the two provinces. Cross‐validation techniques, such as leave‐one‐out and split‐sample, were applied to the associated discriminant functions for each model to check their performance.
• 4 Even though differences existed with respect to the insect composition of the regions, the second model correctly classified 78.1% of the sampled blocks under the non‐organic and organic farming systems at the same time as taking into account two orders: Coleoptera and Hemiptera [excluding Euphyllura olivina olivina (Psyllidae) and the Heteroptera suborder]. The results suggest that the relative abundance of these groups in the post‐blooming period could constitute a potential bio‐indicator of organic olive farming system.

     

Enhancing effects of the chemical adjuvant levamisole on the DNA vaccine pVIR‐P12A‐IL18‐3C

DNA‐based vaccination is an attractive alternative for overcoming the disadvantages of inactivated virus vaccines; however, DNA vaccines alone often generate only weak immune responses. In this study, the efficacy of LMS as a chemical adjuvant on a DNA vaccine (pVIR‐P12A‐IL18‐3C) encoding the P1‐2A and 3C genes of the FMDV and swine IL‐18, which provides protection against FMDV challenge, was tested. All test pigs were administered booster vaccinations 28 days after the initial inoculation, and were challenged with 1000 ID₅₀ FMDV O/NY00 20 days after the booster vaccination. Positive and negative control groups were inoculated with inactivated virus vaccine and PBS respectively. The DNA vaccine plus LMS induced greater humoral and cell‐mediated responses than the DNA vaccine alone, as evidenced by higher concentrations of neutralizing and specific anti‐FMDV antibodies, and by higher concentrations of T‐lymphocyte proliferation and IFN‐γ production, respectively. FMDV challenge revealed that the DNA vaccine plus LMS provided higher protection than the DNA vaccine alone. This study demonstrates that LMS may be useful as an adjuvant for improving the protective efficiency of DNA vaccination against FMDV in pigs.     

Semi-Mechanized Simultaneous Synthesis of Multiple Oligonucleotide Fragments

Abstract

A new strategy for the simultaneous synthesis of multiple oligonucleotides is described, which is based on asynchronous chain growth and use of a maximum number of support segments in each elongation. An approach to the mechanization of this strategy and applications to the preparation of gene fragments and sequentially variant probes are discussed.     

Potential of high‐density pheromone‐releasing microtraps for control of codling moth Cydia pomonella and obliquebanded leafroller Choristoneura rosaceana

Recent large‐cage studies with codling moth Cydia pomonella (L.) reveal that the removal of moths from an apple orchard using pheromone‐releasing traps is more effective at reducing capture in a central monitoring trap than is a mating disruption protocol without kill/capture. The present study uses open orchard 0.2‐ha plots comparing a high‐density trapping scenario with mating disruption to confirm those results. Two tortricid moth pests of tree fruit are studied: codling moth and obliquebanded leafroller Choristoneura rosaceana (Harris). Codling moth treatments include Isomate CM FLEX (ShinEtsu Ltd, Japan), nonsticky traps baited with Trécé CM lures (Trécé, Inc., Adair, Oklahoma), and sticky traps baited with Trécé CM lures, all at equal application rates of 500 dispensers ha^?1, as well as a no pheromone control. These microtraps are of a novel design, small and easy to apply, and potentially inexpensive to produce. Mating disruption using Isomate CM FLEX and nonsticky traps reduces codling moth capture in standard monitoring traps by 58% and 71%, respectively. The attract‐and‐remove treatment with sticky traps reduces capture by 92%. Obliquebanded leafroller treatments include Isomate OBLR/PLR Plus and Pherocon IIB microtraps baited with Trécé OBLR lures, both applied at 500 dispensers ha^?1, as well as a no pheromone control. Mating disruption reduces capture in monitoring traps by 69%. The attract‐and‐remove treatment reduces capture by 85%. Both studies suggest that an attract‐and‐remove approach has the potential to provide superior control of moth populations compared with that achieved by mating disruption operating by competitive attraction.     

Effect of cropping systems on cereal stemborers in the cool‐wet and semi‐arid ecozones of the Amhara region of Ethiopia

1 Field experiments were conducted on maize and sorghum at three locations in the Amhara state of Ethiopia to determine the effects of mixed cropping on stemborer infestation, borer natural enemies and grain yields. In the cool‐wet ecozone of western Amhara, sole maize was compared with maize intercropped with faba bean, mustard, potatoes and cowpea. In the semi‐arid ecozone of eastern Amhara, the trial was conducted on both maize and sorghum with the companion crops haricot bean, sesame, cowpea and sweet potatoes. 2 The results showed that the predominant borer species in western and eastern Amhara were, respectively, Busseola fusca and Chilo partellus. In Addis Zemen, western Amhara, maize intercropped with mustard and potatoes had significantly lower pest numbers and percent tunnelling than other intercrops and the maize monocrop during the vegetative stage. In eastern Amhara, the cropping system did not significantly affect pest densities but damage to stem, ear or heads tended to be greatest when cereals were intercropped with sweet potatoes. 3 Parasitism of C. partellus by the braconid Cotesia flavipes was greater on maize than sorghum, and on maize it was greater with sweet potatoes than in other intercrops or sole maize. Cocoon mass number per plant did not vary significantly between treatments. 4 There were significant differences between treatments in yields of both sorghum and maize (per plant and per unit area) with the lowest yields observed when they were intercropped with a tuber crop. 5 The results suggest that simultaneous planting of the crop species selected has little advantage over monocropped maize.