Search PubMed⌕ Search

Biomedical subjects

J Shanks

Publications and source records attributed to J Shanks.

At least 19 recordsLinked to original sources
PubMed2003

The detection of Simian virus 40 in mesotheliomas from New Zealand and England using real time FRET probe PCR protocols.

AIMS: To detect Simian virus 40 (SV40) DNA in mesotheliomas from New Zealand and from England using novel real time FRET probe polymerase chain reaction (PCR) protocols. METHODS: Twenty four mesotheliomas from New Zealand (Central North Island) and 32 mesotheliomas from England (Greater Manchester region) were examined. Two real time FRET probe PCR protocols were optimised and their analytical sensitivity compared using dilutions of SV40 DNA. A conventional SV40 large tumour antigen protocol with detection by probe hybridisation and chemiluminescent Southern blotting was also optimised. RESULTS: Both real time PCR protocols had the same analytical sensitivity, detecting down to 10(-6) pg of SV40 DNA for each reaction, approximately one SV40 copy. All of the 56 mesothelioma samples contained amplifiable beta globin DNA, but none contained amplifiable SV40 DNA with the conventional large T antigen PCR-Southern blotting protocol, or the two real time FRET probe PCR protocols. The positive and negative controls gave the expected results. There was no evidence of inhibition. CONCLUSIONS: There is abundant evidence in the literature for the presence of SV40 in mesotheliomas. However, this study found no evidence of SV40 in mesotheliomas from England and New Zealand. The extensive use of SV40 contaminated polio vaccine in New Zealand does not seem to have resulted in SV40 associated mesotheliomas.

DNA, Viral↗
PubMed1998

Microtubule dynamic instability does not result from stabilization of microtubules by tubulin-GDP-Pi subunits.

The proposal that microtubule dynamic instability results from stabilization of microtubule ends by tubulin-GDP-Pi subunits (where Pi is inorganic phosphate) [Melki et al. (1996) Biochemistry 35, 12038] was based on studies of GTP hydrolysis and microtubule assembly that showed that tubulin-GDP-Pi subunits can transiently accumulate at microtubule ends. There is no direct evidence that GDP-Pi-subunits can stabilize microtubules under conditions where dynamic instability is observed and this has been inferred from the observation that tubulin-GDP-BeFn subunits stabilize microtubules. To test if tubulin-GDP-Pi stabilizes microtubules we sought evidence for a synergism between the effect of Pi and BeFn. We found, however, that Pi antagonizes the effect of BeFn by displacing it from tubulin subunits. The alternate mechanism in which Pi inhibits BeFn stabilization of microtubules by displacing fluoride from beryllium was ruled out from the 9Be and 19F NMR spectra in the presence and absence of Pi. Further evidence that tubulin-GDP-BeFn is not an analogue of tubulin-GDP-Pi and that tubulin-GDP-Pi is not responsible for maintaining the growth phase in microtubules manifesting dynamic instability was provided by our observation that Pi did not decrease the disassembly rate under conditions where tubulin-GDP-Pi subunits are expected to have formed. Results showing that BeFn binds randomly to subunits in microtubules provided evidence that Pi dissociation from the tubulin-GDP-Pi intermediate formed during GTP hydrolysis occurs randomly rather than processively starting at the growing microtubule tip.

Animals↗
PubMed1998

Opium alkaloid noscapine is an antitumor agent that arrests metaphase and induces apoptosis in dividing cells.

An alkaloid from opium, noscapine, is used as an antitussive drug and has low toxicity in humans and mice. We show that noscapine binds stoichiometrically to tubulin, alters its conformation, affects microtubule assembly, and arrests mammalian cells in mitosis. Furthermore, noscapine causes apoptosis in many cell types and has potent antitumor activity against solid murine lymphoid tumors (even when the drug was administered orally) and against human breast and bladder tumors implanted in nude mice. Because noscapine is water-soluble and absorbed after oral administration, its chemotherapeutic potential in human cancer merits thorough evaluation.

Animals↗
PubMed1997

Regional and fiber-type percentages and sizes in the hamster diaphragm after swim training.

BACKGROUND AND PURPOSE: The purpose of this study was to determine the effect of a swimming endurance training program on changes in percentages and sizes of fiber types in different regions of the hamster diaphragm. METHODS: Adult male golden Syrian hamsters were randomly assigned to a control group (n = 9) or a swimming group (n = 10). Hamsters in the swimming group swam for 80 minutes per session, 5 days per week, for 13 weeks. Fiber-type percentages and sizes were determined for the costal region and for the abdominal and thoracic surfaces of the crural region of the diaphragm from cross sections processed for myofibrillar adenosine triphosphatase. RESULTS: Muscle fibers in the thoracic surface of the crural region were smaller in the swimming group than in the control group. Fiber-type percentages in the diaphragm, however, were not different between groups. CONCLUSION AND DISCUSSION: Swim training may have improved the endurance of the thoracic/crural region by decreasing cross-sectional area and thus decreasing the distance for oxygen to diffuse to the internal regions of the muscle fibers.

Adenosine Triphosphatases↗
PubMed1996

Evidence that a single monolayer tubulin-GTP cap is both necessary and sufficient to stabilize microtubules.

Evidence that 13 or 14 contiguous tubulin-GTP subunits are sufficient to cap and stabilize a microtubule end and that loss of only one of these subunits results in the transition to rapid disassembly(catastrophe) was obtained using the slowly hydrolyzable GTP analogue guanylyl-(a,b)-methylene-diphosphonate (GMPCPP). The minus end of microtubules assembled with GTP was transiently stabilized against dilution-induced disassembly by reaction with tubulin-GMPCPP subunits for a time sufficient to cap the end with an average 40 subunits. The minimum size of a tubulin-GMPCPP cap sufficient to prevent disassembly was estimated from an observed 25- to 2000-s lifetime of the GMPCPP-stabilized microtubules following dilution with buffer and from the time required for loss of a single tubulin-GMPCPP subunit from the microtubule end (found to be 15 s). Rather than assuming that the 25- to 2000-s dispersion in cap lifetime results from an unlikely 80-fold range in the number of tubulin-GMPCpP subunits added in the 25-s incubation, it is proposed that this results because the minimum stable cap contains 13 to 14 tubulin-GMPCPP subunits. As a consequence, a microtubule capped with 13-14 tubulin-GMPCPP subunits switches to disassembly after only one dissociation event (in about 15 s), whereas the time required for catastrophe of a microtubule with only six times as many subunits (84 subunits) corresponds to 71 dissociation events (84-13). The minimum size of a tubulin-GMPCPP cap sufficient to prevent disassembly was also estimated with microtubules in which a GMPCPP-cap was formed by allowing chance to result in the accumulation of multiple contiguous tubulin-GMPCPP subunits at the end, during the disassembly of microtubules containing both GDP and GMPCPP. Our observation that the disassembly rate was inhibited in proportion to the 13-14th power of the fraction of subunits containing GMPCPP again suggests that a minimum cap contains 13-14 tubulin-GMPCPP subunits. A remeasurement of the rate constant for dissociation of a tubulin-GMPCPP subunit from the plus-end of GMPCPP microtubules, now found to be 0.118 s-1, has allowed a better estimate of the standard free energy for hydrolysis of GMPCPP in a microtubule and release of Pi: this is +0.7 kcal/mol, rather than -0.9 kcal/mol, as previously reported.

Animals↗
PubMed1996

Relationship between feeding pattern and body mass index in 220 free-living people in four age groups.

OBJECTIVE: To assess the relationship between feeding pattern and body mass index in free-living humans. DESIGN AND SUBJECTS: Feeding pattern was assessed from 220 7-day weighed dietary records. 187 records were obtained from three separate existing studies, and reanalysed. These studies contained data on three age groups in the British population; Elderly group (n = 88), Middle-aged group (n = 40), Working age group (n = 59). A separate study of 13-14 year olds living in Croydon was conducted from which 33 usable diet records were collected to produce a fourth, Adolescent group. RESULTS: 'Nibbling' and greater energy intakes at breakfast were associated with a lower body mass index (BMI) in the Adolescent group. In the Middle-aged group, greater energy intakes at breakfast and lower energy intakes during the evening were associated with a lower BMI. However, when diet records which produced unreasonably low energy intakes were removed from the analysis, these relationships disappeared except for energy intakes at breakfast and BMI in the Adolescent group. CONCLUSION: It is suggested that the relationship between feeding pattern and BMI observed in the Adolescent and Middle-aged groups was caused by underestimation of 'habitual' energy intake from snacks and the omission of breakfast by females and those who were overweight. The lack of relationship in the Working age group was attributed to the fact that more individuals in this group appeared to report valid diet records. Reported energy intake was directly related to BMI in the Working age group, but was not related to BMI in the other three age groups. It is concluded that feeding pattern is not a major factor in determining BMI in humans. Also, since snacks have a relatively high sugar and low fat composition compared with meals, it is suggested that biased under-reporting of snacks by the obese could produce spurious results from free-living studies which show that obesity is related to the proportion of energy from fat in the diet.

Adolescent↗
PubMed1995

Induction of microtubule catastrophe by formation of tubulin-GDP and apotubulin subunits at microtubule ends.

The recent discovery that GTP linked to latex beads binds to microtubule ends suggested that nucleotide interactions at this site may play a role in regulating microtubule (MT) dynamics. Evidence for this was sought using DIC microscopy to analyze effects of the free GTP and GDP concentration on the rates of MT elongation and phase transition to rapid shortening (catastrophe, kc). That nucleotide can dissociate and thereby destabilize the plus end by forming nucleotide-free (apotubulin) subunits was indicated by an increase in kc from 0.001 to 0.05 s-1, when the free GTP concentration was reduced from 100 to 0.5 microM, during assembly with 15 microM tubulin--GTP subunits (TuT). That nucleotide can bind to the minus end was indicated by a nearly 5-fold decrease in the rate of elongation when the free GDP concentration was increased from 1.6 to 175 microM, during assembly with a mixture of 36 microM TuT and 54 microM TuD. Further evidence that nucleotide can bind to both ends was provided by the observation that with a mixture of 36 microM TuT and 54 microM TuD, kc was increased from 0.0036 to 0.05 s-1 at the plus end, and from 0.0005 to 0.005 s-1 at the minus end, when the free GDP concentration was increased from 1.6 to 175 microM. Our evidence for destabilization of microtubules by formation of apotubulin and by nucleotide exchange to form terminal TuD subunits suggests that microtubule dynamics can be regulated in cells by an exchange factor that generates apotubulin subunits, or by a GTPase activating protein that forms TuD subunits at microtubule ends.

Animals↗
PubMed1995

Analysis from inner London of deprivation payments based on enumeration districts rather than wards.

OBJECTIVE: To estimate the effect of calculating the Jarman index using the smaller geographical unit of the census enumeration district on the changes in deprivation payments made to general practitioners. The Jarman index, or underprivileged area score, is used to calculate the allowance that general practices in the United Kingdom receive for each patient registered with them who lives in an area of relative social deprivation. Current values of the Jarman score are derived from the 1981 census and are based on electoral wards. The change in payments to some practices brought about by using data from the 1991 census may cause severe financial hardship. DESIGN: Jarman indices for wards and enumeration districts from the 1981 and 1991 censuses were used to calculate the payments made to 169 practices in Lambeth, Southwark, and Lewisham; the changes in payments under ward and enumeration district based schemes were then compared. MAIN OUTCOME MEASURES: Standard deviations of the changes in payments to practices. Extreme values of changes in payments. RESULTS: The standard deviation of the change in payment between the two censuses was 6365 pounds with the enumeration district Jarman index, whereas it was 9452 pounds under the ward based scheme. If the ward based scheme is used 10 practices would find their payments changed by over 20,000 pounds, whereas only two practices would have changed by more than this amount under the scheme based on enumeration districts. CONCLUSION: The Jarman index could be more sensitively and appropriately applied to calculate the deprivation payments that practices receive using the census enumeration district as its unit for calculation. This would result in fewer precipitate changes in payments when census data change every 10 years.

Catchment Area, Health↗