p53 mutation, deprivation and poor prognosis in primary breast cancer.

BACKGROUND: The deprivation gap for breast cancer survival remains unexplained by stage at presentation, treatment, or co-morbidities. We hypothesised that p53 mutation might contribute to the impaired outcome observed in patients from deprived communities. METHODS: p53 mutation status was determined using the Roche Amplichip research test in 246 women with primary breast cancer attending a single cancer centre and related to deprivation, pathology, overall, and disease-free survival. RESULTS: p53 mutation, identified in 64/246 (26%) of cancers, was most common in 10 out of 17 (58.8%) of the lowest (10th) deprivation decile. Those patients with p53 mutation in the 10th decile had a significantly worse disease-free survival of only 20% at 5 years (Kaplan-Meier logrank chi(2)=6.050, P=0.014) and worse overall survival of 24% at 5 years (Kaplan-Meier logrank chi(2)=6.791, P=0.009) than women of deciles 1-9 with p53 mutation (c.f. 56% and 72%, respectively) or patients in the 10th decile with wild-type p53 (no disease relapse or deaths). CONCLUSION: p53 mutation in breast cancer is associated with socio-economic deprivation and may provide a molecular basis, with therapeutic implications, for the poorer outcome in women from deprived communities.

Discriminators between hantavirus-infected and -uninfected persons enrolled in a trial of intravenous ribavirin for presumptive hantavirus pulmonary syndrome.

To provide a potentially therapeutic intervention and to collect clinical and laboratory data during an outbreak of hantavirus pulmonary syndrome (HPS), 140 patients from the United States with suspected HPS were enrolled for investigational intravenous ribavirin treatment. HPS was subsequently laboratory confirmed in 30 persons and not confirmed in 105 persons with adequate specimens. Patients with HPS were significantly more likely than were hantavirus-negative patients to report myalgias from onset of symptoms through hospitalization, nausea at outpatient presentation, and diarrhea and nausea at the time of hospitalization; they were significantly less likely to report respiratory symptoms early in the illness. The groups did not differ with regard to time from the onset of illness to the point at which they sought care; time from onset, hospitalization, or enrollment to death was significantly shorter for patients with HPS. At the time of hospitalization, patients with HPS more commonly had myelocytes, metamyelocytes, or promyelocytes on a peripheral blood smear, and significantly more of them had thrombocytopenia, hemoconcentration, and hypocapnia. Patterns of clinical symptoms, the pace of clinical evolution, and specific clinical laboratory parameters discriminated between these 2 groups.

Intravenous ribavirin for hantavirus pulmonary syndrome: safety and tolerance during 1 year of open-label experience. Ribavirin Study Group.

Intravenous ribavirin was provided non-selectively for investigational open-label use among persons with suspected hantavirus pulmonary syndrome (HPS) in the United States between 4 June 1993 and 1 September 1994. Therapy was initiated prior to laboratory confirmation of hantavirus infection because most deaths from HPS occur within 48 h of hospitalization. Thirty patients with confirmed HPS, 105 patients without HPS and 5 patients without adequate diagnostic testing for HPS were enrolled. This observational study arguably provides the most complete information available on ribavirin-associated adverse effects. Although ribavirin was generally well tolerated, 71% of recipients became anaemic and 19% underwent transfusion. An apparent excess of hyperamylasaemia/pancreatitis was either therapy-associated or due to enrollment bias. The 30 enrolled HPS patients had a case-fatality rate of 47% (14/30). It is not possible to assess efficacy with this study design. However, comparison of survival curves for the 30 enrolled HPS patients and 34 patients who developed HPS during the same time period but were not enrolled did not suggest an appreciable drug effect. A randomized, placebo-controlled trial that enrolls patients during the prodrome phase would be necessary to assess the efficacy and further define the safety of intravenous ribavirin for HPS.

Time course of the effects of 6-hydroxydopamine on catecholamine-containing neurones in rat hypothalamus and striatum.

1. The effects of intraventricular injection of 6-hydroxydopamine (6-OHDA) on tyrosine hydroxylase activity, uptake of (3)H-noradrenaline and endogenous catecholamine concentration in rat hypothalamus and striatum were investigated at various times after the injection of 6-OHDA.2. In the hypothalamus after the injection of 250 mug of 6-OHDA there was a rapid decrease in tyrosine hydroxylase activity, (3)H-noradrenaline uptake and noradrenaline content, which was essentially complete within 2 hours.3. In the striatum after this dose of 6-OHDA there was a much slower reduction in tyrosine hydroxylase activity and (3)H-noradrenaline uptake during the first 48 h after drug injection. For the first 24 h the dopamine concentration in this brain area was increased significantly above control values, but had fallen below control values by 48 hours.4. After the injection of a smaller dose of 6-OHDA (25 mug) the only detectable change in the striatum was a rapid increase in the dopamine concentration. In the hypothalamus this dose induced a rapid depletion of noradrenaline, not accompanied initially by any significant reduction in tyrosine hydroxylase activity.5. These results are consistent with the hypothesis that 6-OHDA causes a rapid degeneration of catecholamine-containing nerve terminals in the central nervous system (CNS). These degenerative changes, indicated by the loss of tyrosine hydroxylase and noradrenaline uptake sites, did not appear to be preceded by an initial displacement of endogenous catecholamines by 6-OHDA, except possibly at early times after the administration of small doses of the drug.

Organ culture of Rhipicephalus appendiculatus with maturation of Theileria parva in tick salivary glands in vitro.

A technique is described for the organ culture of Rhipicephalus appendiculatus ticks. Whole, unfed adult ticks with the dorsal integument removed, known as backless tick explants, were cultured in enriched Leibovitz' L-15 medium in which they remained active for at least 32 days at 28 degrees C and 9 days at 36 degrees C. Development of Theileria parva, as demonstrated by methyl green-pyronin staining, occurred in the salivary glands of infected backless tick explants held for 8 days at 28 degrees C or 3 days at 36 degrees C. Maturation in vitro of T. parva in backless tick explants was compared with that in cultured excised salivary glands. After 3-7 days at 36 degrees C glands from backless tick explants and excised salivary glands showed similar numbers of infected acini per infected tick. However, after 12 days at 28 degrees C backless tick explants showed 20-30 times as many infected acini per infected tick as excised salivary glands, in two experiments. No assessment was made of degree of parasite maturity or infectivity. It was concluded that both organ culture techniques supported development in vitro of the salivary gland stages of T. parva, but the backless tick explant technique was simpler and gave generally better results than culture of excised salivary glands.

Alcoholism and cancer. I. Effects of long-term exposure to alcohol on spontaneous mammary adenocarcinoma and prolactin levels in C3H/St mice.

Serum prolactin levels were significantly lower in mice exposed to alcohol for 28 days than they were in controls, and the latency period for adenocarcinoma development was significantly shorter in the alcohol-exposed animals.

Artificial infection of the tick Rhipicephalus appendiculatus with Theileria parva.

Methods for infecting Rhipicephalus appendiculatus ticks with Theileria parva by injection and by artificial feeding were confirmed and compared. The injection method proved simpler and at best as effective and suggested improvements are described.

Daily online bony correction is required for prostate patients without fiducial markers or soft-tissue imaging.

AIM: To compare online position verification strategies with offline correction protocols for patients undergoing definitive prostate radiotherapy. MATERIALS AND METHODS: We analysed 50 patients with implanted fiducial markers undergoing curative prostate radiation treatment, all of whom underwent daily kilovoltage imaging using an on-board imager. For each treatment, patients were set-up initially with skin tattoos and in-room lasers. Orthogonal on-board imager images were acquired and the couch shift to match both bony anatomy and the fiducial markers recorded. The set-up error using skin tattoos and offline bone correction was compared with online bone correction. The fiducial markers were used as the reference. RESULTS: Data from 1923 fractions were analysed. The systematic error was ≤1 mm for all protocols. The average random error was 2-3mm for online bony correction and 3-5mm for skin tattoos or offline-bone. Online-bone showed a significant improvement compared with offline-bone in the number of patients with >5mm set-up errors for >10% (P<0.001) and >20% (P<0.003) of their fractions. CONCLUSIONS: Online correction to bony anatomy reduces both systematic and random set-up error in patients undergoing prostate radiotherapy, and is superior to offline correction methods for those patients not suitable for fiducial markers or daily soft-tissue imaging.

Implementation of an image-guided radiation therapy program: Lessons learnt and future challenges.

The aim of this paper is to detail the experience obtained in implementing an image-guided radiation therapy program at the Northern Sydney Cancer Centre. This required retrofitting a Varian Clinac 21EX with an on-board imager. The commissioning and quality assurance procedures, organisation of a multidisciplinary image guided radiation therapy group, and the development of clinical protocols for orthogonal kV and cone beam computed tomography implementation are described. Reassessment of the image-guided radiation therapy program has continued as new equipment and software versions were made available in the department.

Importance of daily electronic portal imaging in radiotherapy.

An audit was conducted on 20 randomly selected patients who had daily electronic portal imaging during the course of their radiotherapy treatment. The daily images were reviewed to determine whether they were within tolerance according to departmental protocol. If they were not, the actions that were taken were documented. Four treatment areas (spine, chest, breast and prostate) were compared among five patients belonging to each of these categories. The patients were also categorized according to their treatment intent (radical or palliative). A total of 889 electronic portal images of 475 fractions were audited and 33.5% of all fractions were outside tolerance. It was found that 95% of patients needed an action during their treatment and 80% of the patients needed a treatment centre move during the course of their treatment. We found that errors occurred throughout the treatment and it was not possible to predict patients who could have daily imaging omitted. Concordance between radiation therapists and radiation oncologists for identification of error was also investigated. Despite the use of familiar electronic portal imaging protocols, image reviewers (radiation therapists and radiation oncologists) disagreed in interpretation 10% of the time. Our results support the hypothesis that daily imaging may be a useful tool for patients undergoing radiotherapy and that imaging may be ideally carried out before each fraction. Image assessments would be ideally carried out by a team approach, with all images reviewed by both radiation therapists and radiation oncologists. This approach has significant resource implications and may require review of current Medicare and Health Program Grant reimbursements.

The development of Theileria annulata in the salivary glands of the vector tick Hyalomma anatolicum anatolicum.

The development at 28 degrees C of Theileria annulata (Hissar) in the salivary glands of its tick vector, Hyalomma anatolicum anatolicum, was studied using Giemsa-stained smears, methyl green-pyronin-stained preparations of whole glands, and electron microscopy. Nymphs which had engorged on T. annulata-infected calves showed kinetes in the haemolymph from Day 7 to Day 14 post engorgement, when all ticks had completed moult. Intracellular sporonts were observed within salivary gland acini from Day 7 onwards and these developed by rapid nuclear division and cytoplasmic proliferation to form primary sporoblasts. Further development was stimulated by feeding on a rabbit or by incubation at 36 degrees C. The primary sporoblasts appeared to become organized into membrane-bound subunits. Within 48 hours of attachment to the host, or 72 hours of 36 degrees C incubation, these units dissociated to form secondary sporoblasts. The final phase of development resulted in the progressive formation of discrete, uninuclear sporozoites within these secondary sporoblasts. No morphological differences were observed in parasite maturation between the fed and incubated groups although development was retarded by at least 24 hours in the latter. In the incubated group there was also a marked decrease in the degree of synchrony of development which resulted in fewer sporozoites being present at any one time.

A study of the acceptability of lateral records by the Whip-Mix articulator.

1. Of 50 lateral condylar settings made, 10 were not accepted by the Whip-Mix articulator. 2. An improved method of obtaining lateral interocclusal records was described. 3. The Whip-Mix articulator is considered useful in diagnosis and treatment in prosthodontic procedures if one is aware of its limitations.

Bond strengths between elastomeric impression materials and disinfected preliminary impressions.

Bond strengths between disinfected putty impressions and subsequent wash applications were determined in specimens made to simulate clinically an interrupted two-step impression technique. The effect of a disinfectant on the bond strength between a disinfected wash impression and a subsequent wash application was also evaluated to simulate perfecting a disinfected final impression clinically. Vinyl polysiloxane (putty or wash) and polyether (wash only) impression materials were placed in perforated metal cylinders, allowed to set, and dipped in an iodine or glutaraldehyde disinfectant. Wash material was then applied to the disinfected material by use of another metal cylinder and allowed to set before the specimen was tested in tensile mode. Significant decreases in bond strengths were found in both clinical simulations but were limited to specific impression material and disinfectant combinations.

Rapid quantitative assessment of Theileria infection in ticks.

A simplified method for methyl green pyronin staining is described for Theileria parva and T. annulata in whole salivary glands of Rhipicephalus appendiculatus and Hyalomma anatolicum subspecies respectively. The stain gives results comparable with Feulgen staining and can be used after the ticks have been in cold storage for 3 days. There is considerable variability in the rate and intensity of infection of these ticks with theilerial parasites and it is concluded that the method permits large samples (60 ticks per person per day) to be examined to overcome this variability when assessing infection quantitatively.

Murine susceptibility to mercury. I. Autoantibody profiles and systemic immune deposits in inbred, congenic, and intra-H-2 recombinant strains.

Inbred, congenic, and intra-H-2-recombinant mouse strains were given subcutaneous injections of either 1.6 mg HgCl2/kg body wt or 0.1 ml NaCl thrice weekly for 5-6 weeks. Mercury-treated mice from strains carrying the H-2s haplotype developed antinucleolar antibodies (ANoA), which targeted the 34-kDa nucleolar protein fibrillarin, and in some instances also nucleolar proteins of 60-70 and 10-15 kDa, the latter corresponding to histones. Strains with H-2b and H-2d haplotypes were resistant to induction of ANoA. The susceptibility to development of AnoA/antifibrillarin antibodies (AFA) was mapped to the H-2A-region using intra-H-2-recombinant strains. We were not able to confirm earlier reports that expression of H-2E genes dampens the development of ANoA. Mercury treatment caused a substantial increase in the titer of antichromatin (ACA) and/or antihistone (AHA) antibodies in a fraction of SJL/J, A.SW, A.TH, B10.S, and B10.HTT mice (H-2s), and in A/J (H-2k) mice, whereas mice from the C57BL/6J and C57BL/10J (H-2b), and the DBA and BALB/c (H-2d) strains were low or nonresponders. The development of AHA and ACA could not be linked to the H-2 complex. A significant, substantial increase of granular mesangial and systemic vessel wall IgG deposits occurred in mice with serum ANoA/AFA. However, the B10.S(9R) and B10.HTT strains, which express the H-2E genes, developed only an intermediately increased titer of mesangial IgG deposits. Systemic vessel wall IgG deposits occurred in only 60-80% of the B10.S(9R) mice and in none of the B10.HTT mice. This contrasted with the high titer of mesangial IgG deposits and uniform development of systemic vessel wall IgG deposits observed in B10.S mice not expressing H-2E. Mice with mesangial IgG deposits showed a mild glomerulonephritis. There was no systemic vasculitis. The susceptibility to development of ANoA, AHA, ACA, and systemic, granular IgG deposits in the B10.S strain was influenced by the sex, since males showed less uniform development of these immunopathologic features than females.

Murine susceptibility to mercury. II. autoantibody profiles and renal immune deposits in hybrid, backcross, and H-2d congenic mice.

Inorganic mercury causes systemic autoimmunity and/or immune-complex deposits in strains of mice carrying certain H-2 haplotypes, for example H-2s and H-2d. This study aimed at describing the genetic mechanisms regulating these reactions. Inbred SJL, C57BL/6J (B6), C57BL/10J (B10), and DBA mice, F1(SJL x DBA), F1(SJL x B6), and F2(SJL x B6) hybrids, and mice derived from a backcross of SJL or B6 mice to F1(SJL x B6) hybrids were given subcutaneous injections of either 1.6 mg HgCl2/kg body wt or 0.1 ml NaCl every third day for 6 weeks. SJL mice developed a high titer of serum antinucleolar antibodies (ANoA) of the IgG class targeting the nucleolar protein fibrillarin and a significantly increased titer of IgG and C3 colocalized as granular deposits in the renal mesangium and vessel walls. The B6 and DBA strains lacked ANoA and showed no increase in titers of immune deposits. Nine percent of mercury-treated F1(SJL x DBA) hybrids developed IgG-ANoA which were of a low titer, and only occasional hybrids showed an increased titer of granular mesangial IgG deposits. Mercury treatment induced ANoA of low titer in 41% of F1(SJL x B6) hybrids, and 24% had increased granular mesangial immune deposits. Four of 61 mercury-treated BC-[SJL x F1(SJL x B6)] mice showed ANoA which were of a high titer and targeted the nucleolar protein fibrillarin. ANoA were not found in 55 mercury-treated F2(SJL x B6) hybrids or in 56 mercury-treated mice derived from a backcross of B6 mice to F1(SJL x B6) hybrids. Increased mesangial immune deposits were regularly accompanied by vessel wall deposits in F1- and F2(SJL x B6) hybrids, but only 53% of BC(SJL x F1[SJL x B6]) mice with increased mesangial deposits had vessel wall deposits. Vessel wall immune deposits were only present in mice with increased mesangial deposits. A majority of mice which developed significantly increased titers of mesangial IC deposits showed no ANoA. In conclusion, the susceptibility in SJL mice to develop ANoA during mercury treatment, which has been shown to reside in the H-2A locus, was codominantly inherited in a cross with mice carrying the H-2b and H2d haplotypes. Non-H-2 genes dampened ANoA expression to a degree which varied between the strains. Since renal vessel and mesangial IC deposits developed in backcross mice lacking serum ANoA, these deposits must contain IC not related to fibrillarin-antifibrillarin.(ABSTRACT TRUNCATED AT 400 WORDS)

Arsenic and cancer: effects of joint administration of arsenite and selenite on the genesis of mammary adenocarcinoma in inbred female C3H/St mice.

The joint administration of 2 ppm of arsenic as arsenite and of 2 ppm of selenium as selenite in the drinking water of inbred female C3H/St mice increases the incidence of spontaneous mammary adenocarcinoma significantly over the simultaneous controls and animals receiving 2 ppm As as arsenite or 2 ppm Se as selenite alone, indicating that arsenic abolishes the anticarcinogenic effect of selenium. Arsenite caused a significant increase of the tumor growth rates and raised the incidence of multiple tumors. The tumor growth rates in the As-Se group were also increased, but the incidence of multiple tumors was lower than in the 2 ppm As group.

The maintenance and survival of Theileria annulata in colonies of Hyalomma anatolicum anatolicum.

Procedures are described for the colonization of Hyalomma anatolicum anatolicum ticks using gerbils, rabbits and cattle as hosts. On rabbits H. a. anatolicum undergoes a two-host cycle but methods are described for obtaining either unfed or engorged nymphs. Data are given on the life cycle timings and the numbers and timings of tick feeding regimes. The infection of H. a. anatolicum with two strains of Theileria annulata is described and methods given for monitoring the development and survival of T. annulata in H. a. anatolicum. Data are presented indicating the optimum maintenance conditions for T. annulata in H. a. anatolicum. Piroplasm parasitaemias in cattle greater than 2% gave high infection rates in adult ticks if engorged nymphs were moulted at 28 degrees C for 28 days. The Theileria will survive for ten months in such adults stored at 12 degrees C after moult, and when fed the ticks will produce maximum numbers of sporozoites on the third day of feeding, whatever their age. The moult of engorged nymphs is retarded at 18 degrees C but the Theileria in such ticks will develop normally when the ticks are moulted at 28 degrees C after four months storage at 18 degrees C.