Recent Unpublished U.K. Theses on Birds

BOWLER, J.M. 1996. Feeding Strategies of Bewick's Swans Cygnus columbianus brwickii in Winter. PhD Thesis. University of Bristol.     

Q & A. Philip Ingham

Philip Ingham grew up in Liverpool and graduated from Cambridge University in 1977. He did his D.Phil in Developmental Genetics at Sussex University and postdoctoral work in Strasbourg, France before joining the laboratory of David Ish-Horowicz at the ICRF Mill Hill Laboratories. Here he applied the emerging technique of tissue in situ hybridisation to the analysis of the Drosophila segmentation genes. After a short spell at the MRC Laboratory of Molecular Biology in Cambridge, he rejoined the ICRF as a Research Scientist at the Developmental Biology Unit in Oxford. His group pioneered the analysis of the Hedgehog signalling pathway in Drosophila and in collaboration with the labs of Andy McMahon and Cliff Tabin at Harvard University, discovered the Hedgehog gene family in vertebrates. In 1996 he was appointed Professor of Developmental Genetics at the University of Sheffield where he has established the Centre for Developmental Genetics.     

Jonathon Howard.

Jonathon 'Joe' Howard (Fig. 1) is Group Leader and Director at the Max Planck Institute of Molecular Cell Biology and Genetics; he and his research group moved to Dresden, Germany, in July 2001. Howard received his PhD in neurobiology in 1983 from the Australian National University in Canberra. He did postdoctoral research there and also at the University of Bristol, UK, and at the University of California, San Francisco. In 1989, he joined the faculty at the University of Washington. His book "Mechanics of Motor Proteins and the Cytoskeleton" was published earlier this year. [interview by Mari N. Jensen]     

Intracellular nuclease activities of buffalo rumen bacterial population

Nuclease activities of the predominantly bacterial population obtained from buffalo rumen were investigated. Optimum temperature for hydrolysis of both DNA and RNA was 50°C whereas DNAase activity was observed to be stable up to 50°C, a decrease in RNAase activity was observed even after 40°C. Two pH optima, one at 5.5 and the other at 7.5, were recorded for hydrolysis of DNA. RNAase activity was maximum between pH 6.0 to 7.0. Whereas DNAase activity was stable near its optimum pH, RNAase activity was stable between pH 7.0 to 8.5. Mn²⁺ ions stimulated DNAase activity. It was strongly inhibited by Hg²⁺, Zn²⁺, Pb²⁺ and Ag⁺. RNAase activity was stimulated by Mg²⁺ ions and was strongly inhibited by Hg²⁺, Cu²⁺, Zn²⁺ and Ag⁺. Cysteine hydrochloride and 2-mercaptoethanol stimulated DNAase activity. The activity was strongly inhibited by N-ethylmaleimide, 4-chloromercuribenzoate, 8-quinolinol, iodoacetic acid and 1,10-phenanthroline. RNAase activity was stimulated by cysteine hydrochloride, reduced glutathione and 2-mercaptoethanol and was strongly inhibited by 4-chloromercuribenzoate, 8-quinolinol and 2,2′-bipyridyl. Part of PhD Thesis submitted by the first author to Kurukshetra University.     

Q & A. [interview

Joel Rosenbaum was born and grew up in Massena, New York state, on the St Lawrence River border with Ontario, Canada. He received his undergraduate and PhD degrees from Syracuse University, and a Masters Degree in high school biology teaching at St Lawrence University. His PhD work was done with the protozoologist, George Holz Jr, and his post doctoral research on cilia and flagella was at the University Of Chicago with Frank Child and Hewson Swift. He has been at Yale University for 37 years where he has taught Cell Biology. His research has been on the synthesis and assembly of the proteins of cilia and flagella, showing that the flagellar axoneme assembles at the distal tip and that detachment of the flagella upregulates the genes for flagellar proteins. More recently his group has shown that this tip assembly process is facilitated by a rapid kinesin and cytoplasmic dynein-mediated motility underneath the flagellar membrane called ‘intraflagellar transport’. He is a runner with more than 20 marathons under his belt.     

Effect of indoleacetic acid on the metabolic activity oftaraxacum root segments

Inoubation of thin slioes ofTaraxacum root segments in 3-indoleaoetic acid solutions enhanced the level of total proteins, RNA, DNA and a number of enzymes reaching an optimum at 0.01 mg 1^-1. It has been shown that the auxin promotes the synthesis of DNA, again with an optimum promotion at 0.01 mg 1^-1 IAA. This work was carried out at the Department of Botany, University of Sheffield, Sheffield, England. The author wishes to thank Dr. A. Booth for taking a keen interest in this work.     

Cornelis den hartog: An outstanding aquatic ecologist

A survey is given of the work and life of Cornelis den Hartog up to the date in 1996 at which he retired from his position as a professor at the University of Nijmegen. Cornelis (Kees) den Hartog made important contributions to aquatic ecology in the widest sense,e.g. on brackish water typology, meiofauna (microturbellaria), macroinvertebrates, littoral algae, seagrasses and aquatic macrophytes. He favoured the ecosystem approach in aquatic ecology by studying structure and functioning in an integrated way. He lead 31 students to their doctor's degree (PhD).     

Anya Hurlbert and Matt Ridley. [interview

Matt Ridley, the science writer, is married to Anya Hurlbert, reader in visual neuroscience at the University of Newcastle. She studied physics at Princeton University as an undergraduate, then went to Cambridge University as a Marshall Scholar, where she took a Part III diploma in theoretical physics and an MA in physiology. She returned to the States to do the HST MD-PhD program at Harvard-MIT, studying with Tomaso Poggio for her PhD. Matt trained as a zoologist at Oxford, and after completing his PhD on the mating habits of the pheasant, turned to journalism. They met when, as science editor for The Economist, Matt interviewed Anya, and they married while he was the magazine's Washington correspondent. They moved to England, she to Oxford for postdoctoral research, he to become American editor of The Economist. After writing as a columnist for the Daily Telegraph and other papers, Matt turned to writing books: his titles include The Red Queen, The Origins of Virtue, Genome and Nature via Nurture. They now live in rural Northumberland with their two children.     

Cryopreservation of common carp (Cyprinus carpio L.) sperm I. The importance of oxygen supply

Experiments were carried out on the cryopreservation of common carp ( Cyprinus carpio L.) sperm. The effects of pre-freezing oxygen supply on post-thaw motility and the efficacy of different extenders were studied. Sperm diluents contained DMSO as a cryoprotectant in 10% final concentration. The dilution rate was 1:9 (sperm:diluent). Sperm was diluted and equilibrated (10 min) at 0°C. Sperm was then frozen in plastic straws (0.5 ml) at the following rate: 0°C–4°C: 4°C min⁻¹−4°C–80°C: 11°C min⁻¹ from −80°C, straws were plunged directly into liquid nitrogen (−196°C) for further storage. Frozen samples were thawed in a water bath at 40°C.
The freezability of common carp sperm showing reduced motility (due to suboptimal oxygen supply) after transportation could be restored when 30 min of oxygenation was applied prior to freezing. Highest post-thaw motility (57%, percent of control) was achieved when sperm was diluted with modified Kurokura's 'Extender 2'.     

Cryopreservation of common carp (Cyprinus carpio L.) sperm II. Optimal conditions for fertilization

Experiments were carried out on the cryopreservation of common carp ( Cyprinus carpio L.) sperm. Optimal conditions for fertilization including suitable medium and sperm:egg ratio were determined. Sperm was diluted in modified Kurokura's 'Extender 2'containing DMSO as cyroprotectant in 10% final concentration. The dilution rate was 1:9 (sperm:diluent). Sperm was diluted and equilibrated (10 min) at 2°C. Sperm was then frozen in plastic straws (0.5 ml) at the following rate: 0°C–4°C: 4°C min⁻¹; −4°C–80°C: 11°C min⁻¹; from −80°C they were plunged directly into liquid nitrogen (− 196°C). Frozen samples were thawed in a water bath at 40°C. Fertilization rates achieved were much higher in water than in other solutions. Optimal ratios of frozen sperm:egg:water (1:20:20 in volume) and optimal number of frozen spermatozoa:egg (10⁵ spz: 1 egg) were determined. In such conditions, a strong positive correlation (c =+0.846) was found between the post-thaw motility and the fertility of frozen sperm securing high fertilization (99.6%, percent of control). No significant difference was found between fertilization and hatching rates achieved using frozen-thawed common carp sperm.     

Dr. David Rimm is interviewed by Feras Akbik

Dr. David Rimm, MD PhD, is a professor of Pathology at the Yale University School of Medicine specializing in developing quantitative, diagnostic techniques. His lab recently engineered a fluorescence-based algorithm, Automated Quantitative Analysis (AQUA), to analyze tissue microarrays in the hope of moving toward personalized medicine and diagnoses.     

On a new charophyte from India

Chara indica is described as a new species on morphological and cytological grounds.Part of Ph. D. Thesis of Ranchi University.     

The Arbitrary Indian: The Indian Arts and Crafts Act of 1990

The Arbitrary Indian: The Indian Arts and Crafts Act of 1990. Gail K. Sheffield. Norman: University of Oklahoma Press, 1997. 223 pp.     

A rapid and specific enrichment procedure forHyphomicrobium spp.

An enrichment procedure for the isolation of stalked bacteria of the genusHyphomicrobium is described. The method is based on the use of an organic C₁ compound as carbon and energy source for growth together with anaerobic incubation in the presence of nitrate as an electron acceptor. Optimal conditions for the growth of a number ofHyphomicrobium isolates have been investigated. Applying these conditions,Hyphomicrobium spp. have been enriched from a wide range of natural habitats within 1–2 weeks. This work was supported by a Grant from the Medical Research Fund, University of Sheffield, England, to M.M.A.     

Cryopreservation of sperm from the marine shrimp Sicyonia ingentis

Sperm from a marine shrimp, Sicyonia ingentis, were frozen to -196 degrees C using a variety of cooling rates and cryoprotectants. A cooling rate of 1 degree C/min resulted in minimal cell breakage. Sperm samples were frozen in solutions of known membrane stabilizers--trehalose, sucrose, proline, and glycerol. These compounds were somewhat effective but a dramatic increase in sperm viability was seen when DMSO was present in the freezing medium. Sperm viability was assessed using the in vitro acrosome reaction technique of Griffin et al. (1987). The highest sperm survival (56%) was obtained with samples frozen at 1 degrees C/min in a 5% (v/v) DMSO solution. No decrease in viability was seen in sperm samples stored in liquid nitrogen (-196 degrees C) for 1 month.     

Sperm cryopreservation of the critically endangered olive barb (Sarpunti) Puntiussarana (Hamilton, 1822)

The present study focused on development of a sperm cryopreservation protocol for the critically endangered olive barb Puntiussarana (Hamilton, 1822) collected from two stocks within Bangladesh and reared in the Fisheries Field Laboratory, Bangladesh Agricultural University (BAU). The sperm were collected in Alsever’s solution prepared at 296 mOsmol kg⁻¹. Sperm were activated with distilled water (24 mOsmol kg⁻¹) to characterize motility. Maximum motility (90%) was observed within 15 s after activation, and sperm remained motile for 35 s. Sperm activation was evaluated in different osmolalities and motility was completely inhibited when osmolality of the extender was ?287 mOsmol kg⁻¹. To evaluate cryoprotectant toxicity, sperm were equilibrated with 5%, 10% and 15% each of dimethyl sulfoxide (DMSO) and methanol. Sperm motility was noticeably reduced within 10 min, when sperm were equilibrated with 15% DMSO, indicating acute toxicity to spermatozoa and therefore this concentration was excluded in further trials. Sperm were cryopreserved using DMSO at concentrations of 5% and 10% and methanol at 5%, 10% and 15%. The one-step freezing protocol (from 5 °C to −80 °C at 10 °C/min) was carried out in a computer-controlled freezer (FREEZE CONTROL® CL-3300; Australia) and 0.25-ml straws containing spermatozoa were stored in liquid nitrogen for 7–15 days at −196 °C. The highest motility in thawed sperm 61 ± 8% (mean ± SD) was obtained with 10% DMSO. The fertilization and hatching rates were 70% and 37% for cryopreserved sperm, and 72% and 62% for fresh sperm. The protocol reported here can be useful for hatchery-scale production of olive barb. The use of cryopreserved sperm can facilitate hatchery operations, and can provide for long-term conservation of genetic resources to contribute in the recovery of critically endangered fish such as the olive barb.     

Science Educational Outreach Programs That Benefit Students and Scientists

Both scientists and the public would benefit from improved communication of basic scientific research and from integrating scientists into education outreach, but opportunities to support these efforts are limited. We have developed two low-cost programs—"Present Your PhD Thesis to a 12-Year-Old" and "Shadow a Scientist”—that combine training in science communication with outreach to area middle schools. We assessed the outcomes of these programs and found a 2-fold benefit: scientists improve their communication skills by explaining basic science research to a general audience, and students'' enthusiasm for science and their scientific knowledge are increased. Here we present details about both programs, along with our assessment of them, and discuss the feasibility of exporting these programs to other universities.     

Large-scale preparation of T4 endonuclease VII from over-expressing bacteria

Endonuclease VII is the product of gene 49 of phage T4 and was the first enzyme shown to resolve Holliday structures in vitro [Mizuuchi, K. et al. (1982) Cell 29, 357-365]. Low amounts of the enzyme were originally purified from phage-infected cells [Kemper, B. & Garabett, M. (1981) Eur. J. Biochem. 115, 123-131]. We now report a purification procedure for milligram amounts of cloned endonuclease VII expressed in Escherichia coli with gene 49 under the control of a temperature-inducible promoter on a plasmid system [Tomaschewski, J. (1988) PhD Thesis, University of Bochum, FRG]. The protein was purified 500-fold from crude extracts in five steps with a recovery of 15%. The steps include (a) poly(ethyleneglycol)/dextran two-phase separation; (b) DEAE-cellulose; (c) single-stranded DNA-agarose; (d) Mono-Q and (e) Mono-S chromatography. The final protein was more than 98% pure as estimated from SDS/PAGE analysis. The protein has an apparent molecular mass of 17.8 kDa on SDS-containing polyacrylamide gels and 36 kDa when determined by gel filtration or sedimentation through sucrose gradients in the presence of high salt (600 mM NaCl). In the absence of additional salt, the enzyme has a tendency to aggregate and products of molecular masses differing in steps of about 18 kDa appear on SDS-containing polyacrylamide gels.     

Purification and characterisation of soluble invertases from leaves of Arabidopsis thaliana

Multiple isoforms of -fructofuranosidase (invertase, EC 3.2.1.26) were identified in mature green leaves of the cruciferous plant Arabidopsis thaliana (L.) Heynh. There were four major and one minor isoforms of soluble acid invertase and an additional activity which could be released from the cell wall by buffers of high ionic strength. This study reports the separation and characterisation of three soluble isoforms following ammonium sulphate and polyethylene glycol 6000 precipitations, Concanavalin A, MonoQ ion exchange, Superose 12 sizeexclusion chromatography and chromatofocusing. These isoforms, designated INV1, INV2 and INV3, had isoelectric points of 4.75, 4.70 and 4.65 and a K _m for sucrose of 5, 12 and 5 mM, respectively. Each had a pH optimum of 5.5, exhibited optimal activity at 45 °C and used sucrose as the preferred substrate. All fractions containing these isoforms contained a 52-kDa polypeptide which was specifically detected by immunoblotting with an antibody raised against deglycosylated wheat invertase. The N-terminal amino-acid sequence of this polypeptide was homologous to acid invertases isolated from other plant species. The possible origin of isoforms of soluble acid invertase is discussed.Abbreviations PEG polyethylene glycol - pI isoelectric point - PMSF phenylmethylsulphonyl fluoride We wish to acknowledge the support of the British/Swiss Joint Research Programme and the Sheffield University Research Support Fund. X.T. was in receipt of an Overseas Research Scholarship and a University of Sheffield Research Scholarship. We wish to thank Dr A. Moir for his help in N-terminal amino-acid sequencing.