Antidepressants Cause Bradycardia and Heart Block in GD 13 Rat Embryos In Vitro

This study investigated the effects of a range of antidepressant drugs on the heart of gestation day 13 rat embryos in vitro. The general hypothesis was that the drugs would adversely affect the function of the embryonic heart since they all have some cardiac ion channel blocking activity in addition to their main pharmacological effect on neurotransmitters. The results showed that all the tested drugs caused bradycardia in a generally concentration‐dependent manner. At higher concentrations most of the drugs caused some degree of heart block consistent with sodium channel blockade and some drugs also showed negative inotropy associated with blockade of the L‐type calcium channel. One drug, trazodone, caused arrhythmia consistent with blockade of the hERG (human ether‐a‐go‐go related gene) potassium channel. In general the effects on the embryonic rat heart were only seen at “free drug” concentrations much greater than those likely to occur in pregnant women taking antidepressant medication. The least margin of safety was seen with the tricyclic antidepressants and the serotonin antagonist and reuptake inhibitor trazodone.     

Influence of gibberellin biosynthesis inhibitors on stem elongation and floral initiation on in vitro chicory root explants under dark and light conditions

Root explants of chicory (Cichorium intybus L.) were cultured in vitro under continuous light or darkness. On a standard medium (no plant growth regulators added), flowering-stems were initiated under continuous light while under continuous dark, vegetative-stems were formed. Different types of GA (gibberellin) biosynthesis inhibitors were added to the culture medium. Paclobutrazol and compounds belonging to the group of cyclohexanetriones clearly reduced flowering-stem growth under light conditions and vegetative-stem growth under dark conditions. Under light conditions, flower bud initiation was not affected. These and other results suggest that GA₁ may be synthesized during the in vitro culture period and that it controls flowering-stem growth but not floral initiation.Abbreviations CCC chlormequat chloride - GA gibberellin - LAB 198 999 3,5-dioxo-4-butyryl-cyclohexane carboxylic acid ethyl ester - BAS 111..W 1-phenoxy-3-(1H-1,2,4-triazol-1-yl)-4-hydroxy-5,5-dimethylhexane     

Genomic sequencing by ligation-mediated PCR

Genomic sequencing permits studies of in vivo DNA methylation and protein-DNA interactions, but its use has been limited due to the complexity of the mammalian genome. Ligation-mediated PCR (LMPCR) is a sensitive genomic sequencing procedure that generates high quality, reproducible sequence ladders starting with only 1 μg of uncloned mammalian DNA per reaction. This genomic sequencing procedure can be adapted for various methylation, in vivo footprinting and DNA adduct mapping procedures. We provide a detailed protocol for genomic sequencing by LMPCR and discuss the principles and applications of the method.     

Fungi in Their Own Right

Although fungi have contributed tremendously to understanding biological phenomena common to all eukaryotic organisms, some of their properties testify as to their uniqueness. Among these are growth by apical extension of hyphae, the manufacture of hydrophobins for emergence into the air, and the possession of an extended somatic heterokaryon in basidiomycetes. This justifies studies on the molecular basis of development aimed particularly at this group of organisms, which are of great importance to life on earth and human society.     

Targetting of the N‐terminal domain of the human papillomavirus type 16 E6 oncoprotein with monomeric scFvs blocks the E6‐mediated degradation of cellular p53

The E6 protein of cancer‐associated human papillomavirus type 16 (HPV16) binds to cellular p53 and promotes its degradation through the ubiquitin pathway. In an attempt to identify the regions of E6 that could be targetted for functional inhibition, we generated monoclonal antibodies to the HPV16 E6 oncoprotein (16E6) and analysed their effect on E6‐mediated p53 in vitro degradation. The isolated antibodies recognize the 16E6 oncoprotein expressed in the CaSki carcinoma cell line and strongly inhibit the proteolysis of p53 in vitro by binding specifically to a region of 10 residues located at the N‐terminal end of 16E6. The variable regions of these antibodies were cloned and expressed in E. coli as single chain Fvs (scFvs). Purified scFvs were present in monomeric form and totally abolished 16E6‐mediated p53 degradation by preventing the formation of E6/p53 protein complexes. Our results demonstrate that monovalent binding of scFvs to the N‐terminal end of 16E6 abrogates the biological mechanisms leading to the degradation of p53, and they suggest that this region of 16E6 may be a useful in vivo target for blocking the oncogenic activity of HPV16 E6 protein. Copyright © 1999 John Wiley & Sons, Ltd.     

A Fast Air-dry Dropping Chromosome Preparation Method Suitable for FISH in Plants

Preparation of chromosome spreads is a prerequisite for the successful performance of fluorescence in situ hybridization (FISH). Preparation of high quality plant chromosome spreads is challenging due to the rigid cell wall. One of the approved methods for the preparation of plant chromosomes is a so-called drop preparation, also known as drop-spreading or air-drying technique. Here, we present a protocol for the fast preparation of mitotic chromosome spreads suitable for the FISH detection of single and high copy DNA probes. This method is an improved variant of the air-dry drop method performed under a relative humidity of 50%-55%. This protocol comprises a reduced number of washing steps making its application easy, efficient and reproducible. Obvious benefits of this approach are well-spread, undamaged and numerous metaphase chromosomes serving as a perfect prerequisite for successful FISH analysis. Using this protocol we obtained high-quality chromosome spreads and reproducible FISH results for Hordeum vulgare, H. bulbosum, H. marinum, H. murinum, H. pubiflorum and Secale cereale.     

Results of Blood Transfusions from Donor Rabbits Infected with Eimeria stiedai to Recipient Coccidia-free Rabbits*

Blood, withdrawn by heart puncture, from donor rabbits infected with Eimeria stiedai, was transfused by heart puncture, to uninfected recipient rabbits. Fifty-nine of 76 young coccidia-free recipients, given heparinized whole blood, erythrocytes or leucocytes from donors orally inoculated up to 27 days earlier, developed E. stiedai infections.     

Effect of precipitation variability on net primary production and soil respiration in a Chihuahuan Desert grassland

Precipitation regimes are predicted to become more variable with more extreme rainfall events punctuated by longer intervening dry periods. Water‐limited ecosystems are likely to be highly responsive to altered precipitation regimes. The bucket model predicts that increased precipitation variability will reduce soil moisture stress and increase primary productivity and soil respiration in aridland ecosystems. To test this hypothesis, we experimentally altered the size and frequency of precipitation events during the summer monsoon (July through September) in 2007 and 2008 in a northern Chihuahuan Desert grassland in central New Mexico, USA. Treatments included (1) ambient rain, (2) ambient rain plus one 20 mm rain event each month, and (3) ambient rain plus four 5 mm rain events each month. Throughout two monsoon seasons, we measured soil temperature, soil moisture content (θ), soil respiration (R_s), along with leaf‐level photosynthesis (A_net), predawn leaf water potential (Ψ_pd), and seasonal aboveground net primary productivity (ANPP) of the dominant C₄ grass, Bouteloua eriopoda. Treatment plots receiving a single large rainfall event each month maintained significantly higher seasonal soil θ which corresponded with a significant increase in R_s and ANPP of B. eriopoda when compared with plots receiving multiple small events. Because the strength of these patterns differed between years, we propose a modification of the bucket model in which both the mean and variance of soil water change as a consequence of interannual variability from 1 year to the next. Our results demonstrate that aridland ecosystems are highly sensitive to increased precipitation variability, and that more extreme precipitation events will likely have a positive impact on some aridland ecosystem processes important for the carbon cycle.     

膳食结构对供浆员血浆蛋白及血脂含量的影响研究

目的探讨不同的膳食结构对供浆员血浆蛋白及血脂含量的影响。方法测定400例以牛、羊肉和马铃薯为主要膳食结构的供血浆者和400例以猪肉和马铃薯为主要膳食结构的供血浆者的血浆蛋白和血脂水平;同时统计食用高脂肪膳食后血浆出现乳糜的情况。结果经测定以牛、羊肉和马铃薯为主要膳食结构的供血浆者和以猪肉和马铃薯为主要饮食结构的供血浆者的血浆蛋白和血脂水平相当,两者之间无显著性差异(P>0.05);献浆前1 2 h内食用高脂肪膳食后血浆出现乳糜的概率明显增高,与食用清淡膳食相比具有显著性差异(P<0.05)。结论这两种膳食结构的差异对供浆员血浆蛋白及血脂水平没有明显影响;献浆前12 h内应当避免食用高脂肪类膳食。