Experimental validation of a novel compact focusing scheme for future energy-frontier linear lepton colliders.

A novel scheme for the focusing of high-energy leptons in future linear colliders was proposed in 2001 [P. Raimondi and A. Seryi, Phys. Rev. Lett. 86, 3779 (2001)]. This scheme has many advantageous properties over previously studied focusing schemes, including being significantly shorter for a given energy and having a significantly better energy bandwidth. Experimental results from the ATF2 accelerator at KEK are presented that validate the operating principle of such a scheme by demonstrating the demagnification of a 1.3 GeV electron beam down to below 65 nm in height using an energy-scaled version of the compact focusing optics designed for the ILC collider.

Isolation and characterization of a human homologue of the latrophilin gene from a region of 1p31.1 implicated in breast cancer.

We have identified a region of chromosome 1p31.1 that shows high frequency loss of heterozygosity (LOH) in human breast cancer. This region forms part of a 7 Mb YAC/BAC contig. In order to identify candidate sequences, mutation of which might contribute to the development of disease, we have carried out mapping studies of ESTs localized to 1p31.1. This analysis, coupled with library screening and a modified 5' RACE-PCR strategy, resulted in the identification and characterization of a novel gene (LPHH1) which is located adjacent to the smallest region of overlapping loss (SRO) seen in tumours. The 4209 bp open reading frame of the 7 kb LPHH1 transcript encodes a peptide which shows approximately 65% identity to rat latrophilin, a G-coupled, seven span transmembrane protein, which binds alpha-latrotoxin. In the human sequence, whilst conservation of the transmembrane domain is high, the intra- and extracellular domains show two regions of variable structure, which are presumably generated by alternative splicing. Surprisingly, while expression of the rat gene is tightly restricted to neurological and perhaps some endocrine cells, the human sequence appears to be expressed very widely in all normal tissues tested. Northern and RT-PCR analysis of a panel of tumour cell lines showed that LPHH1 expression was variable, apparently elevated in some lines and absent or markedly reduced in others. Furthermore, characterization of the range of transcripts encoded in a breast tumour cell line, compared to normal breast, suggested that gene product variability was higher in the tumour.

Rare variant (Glu to Lys) in the leucine zipper region of CHOP (GADD153).

CHOP (GADD153) has been shown to be a dominant negative inhibitor of specific transcription factors. Direct sequencing of the gene, amplified from the DNA of a Li-Fraumeni family index case (liposarcoma, breast cancer) revealed a constitutional variant within the coding region. This alteration, though not responsible for the Li-Fraumeni phenotype, resulted in a glutamic acid to lysine switch within the leucine zipper domain, at a residue conserved between CHOP and its potential target molecules and between the human and hamster sequences. The variant created a Taq I restriction fragment length polymorphism (RFLP) facilitating screening. Analysis of 159 breast tumour DNA samples detected two encoding variant alleles (tumour and constitutional DNA).

Genetic and functional studies of a germline TP53 splicing mutation in a Li-Fraumeni-like family.

We report an extensive Li-Fraumeni-like family in which there is an unusual spectrum of tumours at relatively late onset. A germline TP53 splice donor mutation in exon 4 is present in all affected family members available for testing. The mutation abolishes correct splicing of intron 4 and techniques of RT-PCR have identified three different aberrant transcripts from the mutant TP53 allele. Using the yeast functional assay to analyse transcripts in cells from a number of family members with the mutant allele, TP53 appears wild-type. Functional studies have been carried out on cells from patients with and without cancer who carry the germline mutation, and on cells from unaffected individuals from the same family who do not carry the mutation. Using a number of functional endpoints known to distinguish between cells carrying mutant or wild-type TP53 alleles, we were unable to discriminate normal (wt/wt) from heterozygous (wt/mut) cells by lymphocyte apoptosis and fibroblast survival following low dose rate ionising radiation exposure. However germline mutation carriers show increased sensitivity to radiation-induced chromosome damage in the G2 phase of the cell cycle, and decreased transient and permanent G1 arrest. These studies demonstrate the importance of fully characterising the effects of TP53 germline mutations, and may explain some of the phenotypic features of this family.

Genomic structure and expression profile of LPHH1, a 7TM gene variably expressed in breast cancer cell lines.

Gene identification studies, centred on a region of overlapping loss of heterozygosity in breast tumours within band 1p31.1, lead to the characterisation of LPHH1, a novel human 7TM gene. The coding sequence of LPHH1 extends over a 60 kb region and comprises in excess of 28 exons. Alternative splicing occurs minimally at five positions, four of which are within the coding sequence. The fifth region of alternative splicing occurs at the extreme 5' end of the transcript. A clear tissue specific bias in alternative exon selection is observed to some degree at all five positions, including the extreme 5' region, which raises the possibility of multiple and perhaps tissue specific promoters. One such putative promoter region, which appears to be utilised predominantly in breast cancer cells, has been identified. LPHH1 is highly evolutionarily conserved, with the simplest (19 exon) gene product being 95% identical between human and rat. Comparison of the alternatively spliced exons between three species, where data are available, has so far revealed 100% identity in the encoded peptide sequences, suggesting conservation of a functional aspect of this splicing. Gene expression has been observed in all tissues and cell lines tested, with the exception of lymphoblastoid and multiple myeloma lines, where there appears to be only a very low level of transcription. LPHH1 also appears to be downregulated in human bone marrow. These data are consistent with a role for the gene products in adhesion-mediated signalling. Analysis of a panel of breast tumour cell lines revealed that a number apparently overexpressed the gene whilst others showed very low levels of transcription. In one case, the overexpression correlated with a low level increase in gene copy number in the tumour line. In addition to differences in the overall levels of expression, LPHH1 mRNAs were alternatively spliced to varying degrees with shifts in the major gene product to truncated or altered forms in some lines. No somatic LPHH1 mutations were detected through sequence analysis of four primary breast tumours that showed loss of the adjacent 1p31.1 marker D1S207.

Cloning and sequence analysis of an Escherichia coli gene conferring bicyclomycin resistance.

We have cloned and sequenced DNA from Escherichia coli that, when present in a high-copy-number plasmid, confers resistance to the diketopiperazine antibiotic, bicyclomycin (Bc). The DNA includes a 378-amino-acid open reading frame (ORF), disruption of which results in the loss of Bc resistance. This ORF contains the BcR gene. Studies using the minicell expression system reveal that a polypeptide of 31 kDa is produced from this cloned region. The ORF maps at 47.1 min on the E. coli genome map. Sequence comparison between the translated ORF and a protein database reveal between 26.5 and 23.4% aa sequence homology to bacterial transmembrane (TM) proteins including those mediating chloramphenicol (Cm) and tetracycline (Tc) resistance and an arabinose-proton symport protein. Sequence analysis using the Diagon program showed the BcR gene product (BcR) had homology with the N-terminal regions of the CmR and TcR-encoded proteins and weak N-terminal homology with the arabinose-proton symport protein. Hydropathy profiles of the BcR protein and CmR products show a striking similarity, both having twelve predicted TM domains.

The histamine H2 receptor agonist impromidine: synthesis and structure-activity considerations.

Impromidine (1) is a potent and selective histamine H2 receptor agonist and its structure comprises a strongly basic guanidine group containing two different imidazole-containing side chains. In this paper we report the synthesis of analogues in which both of the side chains and the guanidine group are modified and tested as agonists or antagonists at histamine H2 receptors on guinea pig atrium. A protonated amidine group linked by a chain of three carbon atoms to a tautomeric imidazole ring appears to be an essential feature for agonist activity and it is suggested that the second imidazole-containing side chain in impromidine mainly contributes toward affinity for histamine H2 receptors.

Cyanoguanidine-thiourea equivalence in the development of the histamine H2-receptor antagonist, cimetidine.

In the histamine H2-receptor antagonist metiamide (2a) isosteric replacement of thione sulfur (=S) by carbonyl oxygen (=O) or imino nitrogen (=NH) affords the urea 2c and guanidine 2d which are antagonists of decreased potency. The guanidine is very basic and at physiological pH is completely protonated. However, introduction of strongly electronegative substituents into the guanidine group reduces basicity and gives potent H2-receptor antagonists, viz. the cyanoguanidine 2b (cimetidine, "Tagamet") and nitroguanidine 2e. A correspondence between the activity of thioureas and cyanoguanidines is demonstrated for a series of structures 1-4. The close correspondence between cyanoguanidine and thiourea in many physicochemical properties and the pharmacological equivalence of these groups in H2-receptor antagonists leads to the description of cyanoguanidine and thiourea as bioisosteres. Acid hydrolysis of the cyanoguanidine 2b yields the carbamoylguanidine 2f at ambient temperatures and the guanidine 2d at elevated temperatures. Cimetidine is slightly more active than metiamide in vivo as an inhibitor of histamine-stimulated gastric acid secretion and has clinical use in the treatment of peptic ulcer and associated gastrointestinal disorders.

Gas chromatographic determination of guanabenz in biological fluids by electron-capture detection.

A sensitive gas chromatographic method for the determination of guanabenz[(2,6-dichlorobenzylidene)amino]guanidine in urine and plasma was developed. The method depends upon the acid hydrolysis of guanabenz to 2,6-dichlorobenzaldehyde, which has strong electron capturing properties and is volatile enough to be eluted from a gas chromatographic column. Concentrations as low as 0.1 ng of guana-benz/ml can be determined and recovery of the drug from urine and plasma samples is 81.8+/- 5.5% (SD). No interferences arising from plasma, urine, or reagents were encountered. Examples of the application of the method are given.

Accident and emergency services at Auckland hospital.

This is an investigation of the role that the accident and emergency department of the Auckland Hospital plays in the provision of medical care in the Auckland area. It is demonstrated that a significant part of the work that is done in that department could equally well be performed by general practitioners and that there is a need for further education of the public concerning the delivery of health care.

Demographic variables in Auckland medical students.

AIMS: To review the important background details of students admitted to the Auckland School of Medicine over its 25 years. METHODS: Data collected at the time of application on 2448 students who successfully gained entry to the course has been analysed and compared with similar demographic variables in the New Zealand population as a whole, and with some of the findings from a survey performed on the total student population enrolling at Auckland University during 1990. RESULTS: Successful applicants had a mean age of 18.6 years, 39.7% were females and 77% were born in New Zealand. Eighty-three percent were European, 3.6% Maori, 2.4% Pacific Islanders and 10.8% Chinese, Indian or other Asian. Sixty-three percent came from cities over 100,000, 16% being from towns less than 20,000 people. State schools were attended by 77% of entrants and 55% went to single sex schools. University students enrolling in 1990 came from affluent backgrounds with 70% of medical students that year being from socioeconomic levels one and two. Parental occupation was found to clearly influence the career choice of students. CONCLUSIONS: Auckland medical students are predominantly from large cities and affluent backgrounds with only those of European origin being admitted in the same proportion as they occupy in the New Zealand population. The high ratio of Asian students reflects their commitment to senior secondary school studies while the lower admission rate for Maori and Polynesian students is due in part to the large number who leave school without completing their secondary education. The possible impact of these demographic variables on recruitment and loss from the course and on the choice and location of practice is discussed.

Cold water survival suits for aircrew.

Laboratory and sea trials were used to compare the effectiveness of three aircrew exposure garments--the British Mark 10, the United States CWU 21/P, and the Canadian U.VIC. Thermofloat jacket. The first two are waterproof coveralls, whereas the third is a neoprene-lined jacket designed on the basis of the "wet suit" concept. Rectal and skin temperatures, electrocardiograms and other variables were measured while subjects, wearing the suits, were immersed in water at temperatures of 70 degrees C and 10.5 degrees C. The three garments were found to be similar in the degree of thermal protection provided, but the Thermofloat jacket appeared superior in other ways and has the greater potential for development. A previously unreported observation was a marked reduction in core cooling rate after the expected linear fall in core temperature. This has possible implications in the conduct of research in this field.

Photographic memory, money, and liposuction: survey of medical students' wish lists.

OBJECTIVES: To examine whether medical students made fewer altruistic wishes and more money oriented wishes in later years of the medical course than students in earlier years. DESIGN: Anonymous questionnaire survey. SETTING: Auckland University School of Medicine. PARTICIPANTS: 520 medical students from 6 years of the course responded to the questionnaire item "If you had three wishes what would you wish for?" MAIN OUTCOME MEASURES: Proportion of wishes in various categories. RESULTS: The three most popular categories of wishes were happiness (34% of students), money (32%), and altruistic wishes (31%). Rates of altruistic wishes (odds ratio=1.05, 95% confidence interval 0.94 to 1.18; P=0.36) and wishes for money (odds ratio=0.96, 0.86 to 1.08; P=0.52) did not vary over the years of the course. Female medical students were more likely than males to make altruistic wishes (36% v 26%; chi(2)=5.68, P=0. 02), intimacy wishes (25% v 18%; chi(2)=3.74, P=0.05), and happiness wishes (42% v 26%; chi(2)=18.82, P=0.0001). Men were more likely than women to make sexual wishes (5% v 0.8%; chi(2)=7.34, P=0.01). CONCLUSIONS: We found no evidence that students were less altruistic and more money oriented in the later years of the medical course.

Isotopic decay of urinary or plasma 3-methylhistidine as a potential biomarker of pathologic skeletal muscle loss.

BACKGROUND: Skeletal muscle loss accompanying aging or cancer is associated with reduced physical function and predicts morbidity and mortality. 3-Methylhistidine (3MH) has been proposed as a biomarker of myofibrillar proteolysis, which may contribute to skeletal muscle loss. METHODS: We hypothesized that the terminal portion of the isotope decay curve following an oral dose of isotopically labeled 3MH can be measured non-invasively from timed spot urine samples. We investigated the feasibility of this approach by determining isotope enrichment in spot urine samples and corresponding plasma samples and whether meat intake up to the time of dosing influences the isotope decay. RESULTS: Isotope decay constants (k) were similar in plasma and urine, regardless of diet. Post hoc comparison of hourly sampling over 10 h with three samples distributed over 10 or fewer hours suggests that three distributed samples over 5-6 h of plasma or urine sampling yield decay constants similar to those obtained over 10 h of hourly sampling. CONCLUSION: The findings from this study suggest that an index of 3MH production can be obtained from an easily administered test involving oral administration of a stable isotope tracer of 3MH followed by three plasma or urine samples collected over 5-6 h the next day.

SCE induction and harlequin staining in mycoplasma-contaminated Chinese hamster cells.

Chinese hamster V79 and CHO cells infected with Mycoplasma hyorhinis show elevated sister-chromatid exchange (SCE) levels but normal cell proliferation and levels of chromosomal aberrations when compared with uninfected cells. Harlequin staining patterns differ from those seen with uninfected cells at similar levels of bromodeoxyuridine (BrdUrd), indicating that BrdUrd is rapidly depleted from the medium by the mycoplasmal uridine phosphorylase and therefore becomes unavailable over the two cell cycles necessary for harlequin staining. Continuous treatment with the antibiotic minocycline restores the SCE level and harlequin staining to that seen in uncontaminated cells. The results suggest that mycoplasma infection should be suspected if harlequin staining patterns indicate a sudden decrease in incorporation of BrdUrd in cells grown in normal levels of BrdUrd.

Selection of Auckland medical students over 25 years: a time for change?

The selection procedures used in Auckland have been reviewed, and the characteristics of those admitted over 25 years analysed. Students are admitted either as school-leavers, mature entrants, or through an affirmative action scheme. A further small number are admitted as part of overseas development assistance. School-leavers are invited for interview on the basis of their academic achievement. Mature students and the affirmative group must have a minimum acceptable academic standard, with the interview playing a dominant role. Two thousand four hundred and forty-eight students have been admitted. The mean age was 18.6 years, and 39.7% were women. Over one half of the students had a parent who had attended university and 13% had a medical parent. One in ten students failed to complete the course, academic failure and withdrawal being of equal importance. The high loss seen in the affirmative group was due to academic failure and has led to the introduction of extra tuition and support for these students. The emphasis on academic achievement by school-leavers has excluded many applicants with outstanding personal qualities. The academic staff has therefore decided to modify the selection procedure, the final rank order of these applicants being based on their personal attributes and life experiences.