Conducting clinical research in the new NHS: the model of cancer. United Kingdom Coordinating Committee on Cancer Research.

The United Kingdom Coordinating Committee on Cancer Research represents the major organizations funding cancer research in the United Kingdom. The deliberations of a working party convened by the committee to evaluate recently expressed concerns that the changes in the NHS threaten research, especially clinical trials to evaluate new treatments, are reported. A survey of contributors to trials coordinated by the committee showed that half are now experiencing difficulties in continuing to participate in clinical trials. The two major problems identified were lack of time and of staff, especially for NHS staff in non-teaching hospitals. Recent changes in junior doctors'' hours and proposed reductions in the length of time for training will exacerbate this. It is possible to identify the direct and indirect excess costs of conducting research in the NHS, but currently the mechanism does not exist to designate funds specifically for this purpose. Consultation with the regional directors of research and development confirmed that the service increment for teaching and research is not the solution for this. Proposals are made to secure future clinical research in the NHS, including finance, indemnity, the licensing of new drugs, the greater use of nurse counsellors, and the value of cancer registries.     

Using Parasitic Load to Measure the Effect of Anthropogenic Disturbance on Vervet Monkeys

Vervet monkeys, Chlorocebus pygerythrus, thrive in urban areas of KwaZulu-Natal, South Africa, and present a suitable model to assess parasitic load as a measure of anthropogenic disturbance, such as urbanization. We collected vervet monkey faecal samples from four study sites representing a gradient of land use and urbanization. We assessed faecal parasites using the faecal flotation method calculating eggs per gram and parasite richness. Overall, the more urban vervet monkey populations had a significantly higher parasite richness and abundance. Our study shows the applicability of using parasite load to measure the effect of urbanization on wildlife.     

Influence of deslorelin (GnRH-agonist) implant on plasma progesterone, first wave dominant follicle and pregnancy in dairy cattle

The objectives of this study were to investigate the effect of a synthetic GnRH-agonist (Deslorelin) implant on CL function and follicle dynamics when administered 48 h after PGF2 alpha, in a timed-insemination protocol, and to determine if the incorporation of a Deslorelin implant into a timed-insemination protocol to synchronize ovulation would be beneficial to the establishment of pregnancy. In Experiment 1, 15 non lactating cyclic Holstein cows received Buserelin (8 micrograms, i.m.) on Day-9, Lutalyse (25 mg, i.m.) on Day-2, and then on Day 0 received either a Deslorelin implant (700 micrograms, s.c.; n = 5), Buserelin (8 micrograms, i.m.; n = 5), or no treatment (control; n = 5). Blood samples were collected on Days-9, -2, 0 and thereafter daily until the next ovulation. Ovaries were scanned by ultrasound on Days-9, -2, 0, 1 (day of ovulation) and 3 times a week thereafter until a subsequent ovulation. From Days 0 to 15, the rate of increase of plasma progesterone (P4) was greater (P < 0.01) for Deslorelin than for control and Buserelin. Establishment of the first-wave dominant follicle (FWDF) as a Class 3 (> 9 mm) follicle was delayed (P < 0.01) with Deslorelin (14.2 +/- 1.3 d) compared with the control (4.6 +/- 1.3 d) and Buserelin (5.0 +/- 1.5 d) treatments. The FWDF resumed growth after Day 13 in all 5 Deslorelin-treated cows, and 2 cows ovulated spontaneously. In 1 Deslorelin-treated cow, the FWDF regressed, and a second-wave dominant follicle ovulated, while 2 other Deslorelin cows failed to ovulate until after Day 36. The cumulative numbers of Class 2 and 3 follicles was lowest in the Deslorelin group (P < 0.01), while the cumulative number of Class 1 follicles was highest (Deslorelin > Buserelin > Control; P < 0.01). The number of days to CL-regression and days to subsequent estrus did not differ (P > 0.05) among treatments. In Experiment II, 16 lactating potentially subfertile (body condition score 2.25) cows received Cystorelin (100 micrograms, i.m.; Day-9), Lutalyse (25 mg, i.m.; Day-2), and either a Cystorelin injection (100 micrograms, i.m.; n = 8) or Deslorelin implant (700 micrograms, s.c.; n = 8) on Day 0 and inseminated 16 h later. Deslorelin-treated cows had a higher plasma P4 concentration between Days 0 and 16 (P < 0.05) than the 2 other groups, and 5 of the 8 cows in this group were pregnant (Day 45, palpation) compared with 1 of 8 cows in the Cystorelin group (P < 0.05). Incorporation of a Deslorelin implant into a timed-insemination protocol enhanced the pregnancy rate in cows of poor body condition. The results support the hypothesis that enhanced CL function and delayed establishment of the first-wave dominant follicle may enhance embryo survival.     

<Emphasis Type="Italic">α</Emphasis>-Amylase inhibitor-1 gene from <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> expressed in <Emphasis Type="Italic">Coffea arabica</Emphasis>plants inhibits α-amylases from the coffee berry borer pest

Background  

Coffee is an important crop and is crucial to the economy of many developing countries, generating around US70 billion per year. There are 115 species in the < i > Coffea < /i > genus, but only two, < i > C. arabica < /i > and < i > C. canephora < /i > , are commercially cultivated. Coffee plants are attacked by many pathogens and insect-pests, which affect not only the production of coffee but also its grain quality, reducing the commercial value of the product. The main insect-pest, the coffee berry borer ( < i > Hypotheneumus hampei < /i > ), is responsible for worldwide annual losses of around US70 billion per year. There are 115 species in the Coffea genus, but only two, C. arabica and C. canephora, are commercially cultivated. Coffee plants are attacked by many pathogens and insect-pests, which affect not only the production of coffee but also its grain quality, reducing the commercial value of the product. The main insect-pest, the coffee berry borer (Hypotheneumus hampei), is responsible for worldwide annual losses of around US500 million. The coffee berry borer exclusively damages the coffee berries, and it is mainly controlled by organochlorine insecticides that are both toxic and carcinogenic. Unfortunately, natural resistance in the genus Coffea to H. hampei has not been documented. To overcome these problems, biotechnological strategies can be used to introduce an α-amylase inhibitor gene (α-AI1), which confers resistance against the coffee berry borer insect-pest, into C. arabica plants.     

Effect of an implant containing the GnRH agonist deslorelin on secretion of LH, ovarian activity and milk yield of postpartum dairy cows

Prevention of high plasma progesterone concentrations in the early postpartum period may improve fertility. Our objective was to determine whether a Deslorelin implant (DESL; 2100 microg, s.c.) would reduce secretion of LH and alter follicle dynamics, plasma concentrations of progesterone, estradiol and PGF2alpha metabolite (PGFM) in postpartum dairy cows. Cows received DESL on Day 7 postpartum (Day 7, n=8) or were untreated (Control, n=9). All cows were injected with GnRH (100 microg, i.m.) on Day 14 to assess LH response. A protocol for synchronization of ovulation with timed AI was initiated on Day 60 (GnRH [Day 60], CIDR [Day 60 to Day 67], PGF2alpha [Day 67, 25 mg and Day 68, 15 mg], GnRH [Day 69] , AI [Day 70]). The LH response to injection of GnRH on Day 14 was blocked in animals treated with DESL. Numbers of Class 1 (<6 mm) follicles were unaffected (P > 0.05) whereas numbers of Class 2 (6 to 9 mm) (P < 0.01) and Class 3 (>9 mm) follicles were less (P < 0.01) in DESL cows between Day 7 and Day 21. From Day 22 to Day 60, DESL-treated cows had more of Class 1 follicles and less Class 2 (P < 0.01) and Class 3 (P < 0.01) follicles, and lower plasma concentrations of progesterone and estradiol (P < 0.01). Concentrations of PGFM between Day 7 and Day 42 were not affected by treatment (P > 0.05). All cows ovulated in response to GnRH on Day 69. Subsequent luteal phase increases in plasma progesterone concentrations (Day 70 to Day 84) did not differ. The use of the DESL implant associated with PGF2alpha given 14 days later suppressed ovarian activity and caused plasma progesterone concentrations to remain < 1 ng/mL between Day 22 and Day 51. The DESL implant did not affect milk production.     

Neutral super-oxidised solutions are effective in killing P. aeruginosa biofilms

Bacteria growing in biofilms can become up to 1000-fold more resistant to antibiotics and biocides as compared to their planktonic counterparts. As a result of this increased resistance, biofilms and biofilm-related infections cannot be effectively treated with conventional antibiotic therapy. The goal of this study was to determine the efficacy of three neutral pH, super-oxidised solutions (nSOSs, OIS-80, OIS-125, OIS-200, Microcyn Technology) varying in oxychlorine concentration (80, 125 and 200 ppm) against P. aeruginosa grown planktonically and as biofilms. Exposure for 20 s of exponential phase cells to any of the three solutions was sufficient to reduce viability by more than five logs. However, only exposure for 10 min to OIS-125 and OIS-200 for 10 min was sufficient to eradicate stationary phase P. aeruginosa cells. The efficacy of nSOSs on P. aeruginosa biofilms, grown to maturity in continuous flow tube reactors, was determined upon treatment up to 60 min. Viability pre- and post-treatment was determined by CFU counts. The effect of these solutions on P. aeruginosa biofilms and biofilm architecture was further visualised by confocal scanning laser microscopy and quantitatively analysed by COMSTAT. Under these experimental conditions, only OIS-125 and OIS-200 achieved a >3-log reduction and biofilm disaggregation within 30 min of exposure. Because OIS-125 and OIS-200 enhance the disaggregation of biofilms, their use in the treatment of surface-related biofilm infections deserves further investigation.     

Purification of essentially RNA free plasmid DNA using a modified Escherichia coli host strain expressing ribonuclease A

Regulatory agencies have stringent requirements for the large-scale production of biotherapeutics. One of the difficulties associated with the manufacture of plasmid DNA for gene therapy is the removal of the host cell-related impurity RNA following cell lysis. We have constructed a modified Escherichia coli JM107 plasmid host (JMRNaseA), containing a bovine pancreatic ribonuclease (RNaseA) expression cassette, integrated into the host chromosome at the dif locus. The expressed RNaseA is translocated to the periplasm of the cell, and is released during primary plasmid extraction by alkaline lysis. The RNaseA protein is stable throughout incubation at high pH ( approximately 12-12.5), and subsequently acts to hydrolyse host cell RNA present in the neutralised solution following alkaline lysis. Results with this strain harbouring pUC18, and a 2.4 kb pUC18DeltalacO, show that sufficient levels of ribonuclease (RNase) activity are produced to hydrolyse the bulk of the host RNA. This provides a suitable methodology for the removal of RNA, whilst avoiding the addition of exogenous animal sourced RNase and its associated regulatory requirements.     

Synthesis and oxidative insolubilization of cell-wall proteins during osmotic stress

The cell walls in the new white roots of jack pine (Pinus banksiana Lamb.) were observed to constrict around the shrinking protoplast of osmotically stressed roots, and pressure was maintained via an apparent adjustment of cell-wall size and elasticity. These elastic alterations of the cell wall permitted the root cells to maintain full turgor despite the loss of most of the water in the tissue. The constriction of the root cell wall around the dehydrating protoplasts to maintain turgor may reflect changes in cell wall structure. We found that these shrinking root cells synthesize and secrete into the intercellular fluid a set of proteins. These proteins become tightly associated (i.e. guanidine HCl- and sodium dodecyl sulfate-insoluble) with the cell wall but can be released from the matrix, after briefly boiling in 0.1% sodium dodecyl sulfate, by the combination of guanidine HCl, CaCl₂ and dithiothreitol. However, these cell-wall proteins became insoluble with time. The proteins could subsequently be destructively extracted from the wall with acid NaClO₂ treatments. After these proteins were incorporated into the cell walls, the roots adopted a new, smaller maximal tissue volume and elastic coefficients returned to normal levels. Received: 8 July 1998 / Accepted: 19 November 1998     

A study of prostaglandin F2α as the luteolysin in swine: II characterization and comparison of prostaglandin F, estrogens and progestin concentrations in utero-ovarian vein plasma of nonpregnant and pregnant gilts

Polyvinyl catheters were inserted into the right and left utero-ovarian veins (UOV) and saphenous vein (SV) and artery (SA)_of six nonpregnant (O) and five pregnant (P) gilts on day 11 after onset of estrus. Beginning on day 12, UOV blood samples were collected at 15-min intervals from 0800 to 1100 hr and 2000 to 2300 hr, and single samples were taken at 1200 and 2400 hrs. Peripheral blood (SA or SV) was sampled at 0800, 1200, 2000 and 2400 hr until gilts returned to estrus ( ) or day 24 or pregnancy. UOV plasma PGF concentrations (ng/ml; n = 1929) were measured by RIA. Status (P O) by day interactions were detected (P<.01) but variances among treatments were heterogenous (P<.01). Cuvilinear day trends were detected for PGF in 0 gilts (P<.01) but not P gilts. PGF peaks, defined as concentrations greater than two SD above the mean concentration for each gilt, occurred with greater frequency ((ifχ² = 16.4; P>.01)) in 0 than P gilts; and mean peak levels ( ) were 5.0 ± .27 and 3.84 ± .13 ng/ml, respectively.Progesterone concentrations were maintaiend in pregnant pigs and were indicative of luteal maintenance. Systematic differences in day trends of utero-ovarian venous plasma estradiol were detected between O and P pigs. These differences may be of paramount physiological importance and are discussed.