Angiogenic effects despite limited cell survival of bone marrow-derived mesenchymal stem cells under ischemia.

Bone marrow-derived mesenchymal stem cells (MSCs) are multipotent and secrete angiogenic factors, which could help patients with occlusive arterial diseases. We hypothesize that MSCs, in comparison to fibroblasts, survive better under hypoxic conditions in vitro and in vivo. MSCs and fibroblasts from L2G mice expressing firefly luciferase and GFP were cultured in normoxic and hypoxic conditions for 24 hours. In vitro cell viability was tested by detecting apoptosis and necrosis. MSCs released higher amounts of VEGF (281.1 +/- 62.6 pg/ml) under hypoxic conditions compared to normoxia (154.9 +/- 52.3 pg/ml, p = NS), but were less tolerant to hypoxia (45 +/- 7.9%) than fibroblasts (28.1 +/- 3.6%, p = NS). A hindlimb ischemia model was created by ligating the femoral artery of 18 FVB mice. After one week, 1 x 106 cells (MSCs, fibroblasts or saline) were injected into the limb muscles of each animal (n = 6 per group). Bioluminescence measurement to assess the viability of luciferase positive cells showed significant proliferation of MSCs on day four compared to fibroblasts (p = 0.001). Three weeks after cell delivery, the capillary to muscle fiber ratio of ischemic areas was analyzed. In the MSC group, vessel density was significantly higher than in the fibroblast or control group (0.5 +/- 0.08 and 0.3 +/- 0.03). Under hypoxia, MSCs produced more VEGF compared to normal conditions and MSC transplantation into murine ischemic limbs led to an increase in vessel density, although MSC survival was limited. This study suggests that MSC transplantation may be an effective and clinically relevant tool in the therapy of occlusive arterial diseases.

siRNA silencing of keratinocyte-specific GFP expression in a transgenic mouse skin model.

Small interfering RNAs (siRNAs) can be designed to specifically and potently target and silence a mutant allele, with little or no effect on the corresponding wild-type allele expression, presenting an opportunity for therapeutic intervention. Although several siRNAs have entered clinical trials, the development of siRNA therapeutics as a new drug class will require the development of improved delivery technologies. In this study, a reporter mouse model (transgenic click beetle luciferase/humanized monster green fluorescent protein) was developed to enable the study of siRNA delivery to skin; in this transgenic mouse, green fluorescent protein reporter gene expression is confined to the epidermis. Intradermal injection of siRNAs targeting the reporter gene resulted in marked reduction of green fluorescent protein expression in the localized treatment areas as measured by histology, real-time quantitative polymerase chain reaction and intravital imaging using a dual-axes confocal fluorescence microscope. These results indicate that this transgenic mouse skin model, coupled with in vivo imaging, will be useful for development of efficient and 'patient-friendly' siRNA delivery techniques and should facilitate the translation of siRNA-based therapeutics to the clinic for treatment of skin disorders.

Multiple functions for actin during filamentous growth of Saccharomyces cerevisiae.

Saccharomyces cerevisiae is dimorphic and switches from a yeast form to a pseudohyphal (PH) form when starved for nitrogen. PH cells are elongated, bud in a unipolar manner, and invade the agar substrate. We assessed the requirements for actin in mediating the dramatic morphogenetic events that accompany the transition to PH growth. Twelve "alanine scan" alleles of the single yeast actin gene (ACT1) were tested for effects on filamentation, unipolar budding, agar invasion, and cell elongation. Some act1 mutations affect all phenotypes, whereas others affect only one or two aspects of PH growth. Tests of intragenic complementation among specific act1 mutations support the phenotypic evidence for multiple actin functions in filamentous growth. We present evidence that interaction between actin and the actin-binding protein fimbrin is important for PH growth and suggest that association of different actin-binding proteins with actin mediates the multiple functions of actin in filamentous growth. Furthermore, characterization of cytoskeletal structure in wild type and act1/act1 mutants indicates that PH cell morphogenesis requires the maintenance of a highly polarized actin cytoskeleton. Collectively, this work demonstrates that actin plays a central role in fungal dimorphism.

Pulmonary artery sarcoma mimicking pulmonary embolism.

Two cases of primary pulmonary sarcoma which were initially diagnosed as pulmonary thromboembolism are presented. The clinical and radiologic features of pulmonary sarcoma are reviewed, with special emphasis on the features which distinguish it from thromboembolism.

An audit of clinico-radiologic conferences.

RATIONALE AND OBJECTIVES: To assess the various functions of regular review conferences between clinicians and radiologists, proceedings at 19 such meetings were audited. In particular, the authors attempted to quantify their educational component. METHODS: The cases discussed during each meeting were monitored, and the educational component was assessed. Changes in imaging strategy, diagnosis, and management were noted. The participants were questioned regarding their perceptions of such meetings. RESULTS: Two hundred cases were presented at 19 meetings (mean duration 44.8 minutes). Overall, 75 of the 200 cases (37.5%) were presented for routine purposes, 100 (50%) for radiologic assistance, and 25 (12.5%) for general interest. More changes in diagnosis and management and a greater educational contribution occurred in those cases presented for radiologic assistance than in those presented out of routine. Most participants considered that these meetings bestowed considerable benefit to their continuing medical education and to the management of their patients. Discussion considered of educational value developed in 88 of the 200 patients presented; this discussion required 61.7% of the total duration of these meetings. CONCLUSIONS: These conferences provide benefit to the patient, as well as serving an educational role for the medical community. Discussion of imaging strategies and the presentation of controversial and interesting cases should be particularly encouraged.

Urinary iodine and thyroid status of New Zealand residents.

OBJECTIVES: The aim of this project was to assess the clinical significance of our low iodine excretions in terms of thyroid hormone status and thyroid volume in an adult population in a low soil iodine area of the South Island of New Zealand. DESIGN AND SETTING: Two-hundred and thirty-three residents of Otago, New Zealand collected two 24 h urine samples for assessment of iodine status. Thyroid status was determined from serum total T(4), TSH and thyroglobulin, and thyroid volumes. Relationships between urinary iodide excretion and measures of thyroid status were determined and subjects were allocated to one of three groups according to low, medium and high iodide excretion, for comparison of thyroid hormones and thyroid volumes. RESULTS: Significant correlations were found for relationships between measures of urinary iodide excretion and thyroid volume and thyroglobulin. Multiple regression analysis of data for subjects divided into three groups according to 24 h urinary iodide excretion (<60, 60-90; >90 microg iodide/day) or iodide/creatinine ratio (<40; 40-60; >60 microg/g Cr) showed significant differences in thyroid volume (P=0.029; P=0.035, respectively) and thyroglobulin (P=0.019; P=0.005, respectively) among the groups. CONCLUSIONS: The results of this study confirm the low iodide excretions of Otago residents, and indicate that the fall in iodine status is being reflected in clinical measures of thyroid status, including enlarged thyroid glands and elevated thyroglobulin. Our observations suggest the possible re-emergence of mild iodine deficiency and goitres in New Zealand. This situation is likely to worsen should iodine intakes continue to fall and continued monitoring of the situation is imperative.

Histopathological correlates of abnormal pericolic fat on CT in the assessment of colorectal carcinoma.

The aim of this study was to assess the pathological significance of abnormal pericolic fat shown by CT in the context of colorectal carcinoma. CT and histopathological findings of 63 resected colorectal carcinomas were retrospectively reviewed. CT examinations were assessed by two observers for the presence or absence of abnormal pericolic fat (typically linear or nodular opacities) at tumour sites. Specimens were reviewed histopathologically for depth of tumour invasion, extramuscular tissue reaction, and number and largest size of tumour-involved and tumour-free lymph nodes. The sensitivity, specificity, positive predictive value and negative predictive value of pericolic fat in identifying extension of tumour infiltration beyond the muscle coat were 79% (42/53), 33% (2/6), 91% (42/46) and 15% (2/13), respectively. Despite these indicators of efficacy, the association between the presence of pericolic fat abnormality on CT and extramuscular extension of tumour (infiltration and/or nodal disease) or tissue reaction alone or in combination did not reach statistical significance (p>0.3 in all cases). Abnormal ("misty" or "mucky") pericolic fat in the assessment of colorectal cancer on CT is not a precise indicator of extramuscular extension of tumour, as it cannot clearly distinguish between tumour infiltration and tissue reaction beyond the muscle coat, or pericolic nodal involvement. However, it is a very helpful CT sign that may draw attention to the presence and site of a potential colonic abnormality.

Pneumatosis cystoides gastrica associated with adenocarcinoma of the stomach.

This paper reports the finding of benign foreign-body-type multinucleated giant cells and clumps of adenocarcinoma cells in gastric brushing cytology specimens of a case confirmed by radiology and histology as pneumatosis cystoides gastrica associated with an ulcerated carcinoma. Pneumatosis cystoides gastrica should be included in the differential diagnosis of benign multinucleated giant cells found in gastric cytology smears.

Osteitis--an under-recognised association with seronegative spondyloarthropathy?

AIM: To emphasise osteitis as a feature of the spondyloarthritides. METHOD: We describe four cases spanning a spectrum of the spondyloarthritides in which osteitis was a feature. RESULTS: One patient had psoriatic arthritis with palmar-plantar pustular psoriasis and extensive osteitis involving the tibia and fibula. This case provides a link with two cases with SAPHO syndrome (synovitis, acne, pustulosis hyperostosis, osteitis) who had palmar-plantar pustulosis and osteitis. Many now argue that this syndrome is a form of spondyloarthritis. The fourth case, which was of particular interest to us, had enteric reactive arthritis and scintigraphic changes strongly suggesting the presence of osteitis of individual bones in the wrist. CONCLUSION: We propose that these four cases demonstrate that osteitis may be another feature common to the spondyloarthritides and SAPHO. Awareness of this may facilitate better documentation of this feature of the disease.

Once is enough--why some women do not continue to participate in a breast cancer screening programme.

AIM: To assess the reasons why many women who have been screened once in a breast screening programme decline an invitation for further screening. METHODS: Telephone interview survey of a sample of such women; for questions relating to their experience of previous mammography, comparison to data on a representative sample of first screen attendees. The subjects were women who had attended the first round of the Otago-Southland breast cancer screening programme in 1991-1994, who were eligible for re-screening but had been rescreened; age range 50-62. RESULTS: From programme records, 86% of women who were eligible for a second screen accepted it. Of the women not recorded as having had a second screen, some had attended for a second screen; some had not been invited until they had become age ineligible and some had received no invitation for re-screening. Of women who had received and declined an invitation for re-screening (n = 81), the major reason (46%) was their previous mammogram being painful. Other factors contributing were illness in themselves or their spouse, practical difficulties arranging time and negative experiences with staff in the previous mammography, although these related to relatively few women. A few women thought mammography would be of no benefit, and a few thought re-screening was unnecessary because their first mammography had been normal, or because they practise self-examination. CONCLUSIONS: Ensuring that all women eligible for further screening do get invited could substantially increase the re-screening rate. Even women who have declined previous invitations should be offered further invitations, as a substantial proportion with to be screened. Flexible and convenient appointment times are the main modifiable logistic issue. The major factor influencing non-participation with further screening is a painful experience of mammography. Innovative approaches, either to reduce the pain or to reduce the impact of the pain on the woman's attitude to re-screening, should be trialed.

The general practitioner's role in breast cancer screening: a survey in Otago-Southland.

AIMS: To study the experience of general practitioners in Otago and Southland with the existing breast cancer screening programme and the reviews on future programmes. METHODS: A questionnaire was sent to all 210 general practitioners in Otago and Southland in June 1996. RESULTS: The response rate was 71%. All the 141 respondents except one encouraged eligible women to take part in the programme; this was done mainly during individual doctor-patient consultations, by pamphlets and posters, and in the work of the practice nurse. Ten percent of practitioners had a practice-based recall system for breast cancer screening. Seventy-five percent of general practitioners currently provide a list of eligible women to the programme, and of these, 52% check the list to exclude ineligible women. Only 24% of practitioners supplying a patient list to the programme reported that a patient had ever requested that their name be excluded from the list. Twenty-five percent of general practitioners providing lists had a notice in the waiting room stating that. Of those who did not provide lists, concerns about logistics, ethical issues and cost were raised, although 40% of these general practitioners intended to provide lists in the future. In a future programme, 57% of general practitioners felt they should be paid for supplying lists defined by age only and 82% felt they should be paid for supplying a list of women eligible by both age and medical history. Most general practitioners felt that general practitioner lists were the preferred source for invitations to the breast screening programme and that general practitioners had an important part in any future programme. Screening at the ages 50-64 (as currently proposed) is supported by 95% of general practitioners; in addition, 64% supported screening at ages 65-69. Only a minority of general practitioners supported screening at ages 40-49 or ages 70-74. Most general practitioners would offer screening to women under age 50 with either a strong or a weak family history, or even with a past history of a fibroadenoma. CONCLUSIONS: These results show that almost all general practitioners support breast cancer screening programmes and feel that they have an important role in future programmes. The majority support extension of the programme to ages 65-69, but not to ages 40-49. The majority support screening women with individual risk factors at ages under 50, although their responses show that better information on the importance of different risk factors is required.

Clinical outcomes of the Otago-Southland Breast Cancer Screening Programme 1991-1996.

AIM: To document the clinical outcome of the Otago-Southland Breast Cancer Screening Programme through its first two rounds of screening, from 1991-1996. METHODS: Review and analysis of clinical and pathological records. RESULTS: In the first round of screening, 13,876 women were screened, giving 75% uptake; 12.2% were referred for assessment and 126 cancers detected, 9.1 per thousand women screened. For the 9946 incidence screens in the second round, 3.9% of women screened were referred to assessment and 50 cancers detected, 5.0 per thousand women screened. The uptake and cancer detection rates exceed the targets and exceed other published results; the size distribution of the cancers detected was comparable to the Swedish two-counties study, showing that the results should produce an ultimate mortality reduction. The referral rate to assessment was higher than expected in the first round of screening, but within the targeted range in the second round. The benign to malignant ratio for all biopsies was 1.4:1 for the prevalence screen of the first round and 1.2:1 for the incidence screens in the second round, both exceeding the targets set. CONCLUSIONS: The results show that the uptake and clinical results of the programme exceed expectations and that a large number of small invasive tumours have been successfully detected. These results are comparable to the best of overseas studies, and give confidence that mortality reductions will ultimately occur.

Hemifacial atrophy with bilateral short roots.

We present a case in which the patient had both bilateral shortening of the roots, and hemifacial atrophy. As far as we know, this combination has not been described before.

Minimal-preparation abdomino-pelvic CT in frail and elderly patients: prognostic value of colonic and extracolonic findings.

AIM: To examine the overall survival of patients who had had been referred for minimal preparation abdomino-pelvic computed tomography (MPCT), and to assess the prognostic value of the colonic and extracolonic findings detected. METHODS AND MATERIALS: The survival of a cohort of 1029 elderly and frail patients, with clinical symptoms and signs suspicious for colorectal cancer (CRC), who had undergone MPCT between 1995 and 1998 was investigated. Univariate and multivariate survival analyses were undertaken according to the presence of CRC and extracolonic abnormalities (ECA). RESULTS: The median age of the 1029 patients was 79.4 years. The overall median survival following MPCT was 5.4 years; and 6.6 years if no abnormality was detected. On multivariate analysis, age, sex, CRC status, and number of ECAs were significant factors in overall survival. Median survival for those with confirmed CRC [n=91 (prevalence, 8.8%)] was 1.1 years, compared with 5.9 years without CRC (p<0.0001); and 2.4 years for those with one or more ECA [n=245 (prevalence, 23.8%)], compared with 6.1 years without ECA (p<0.0001). Survival was progressively shorter for increasing numbers of ECAs; and shorter for previously unknown non-CRC malignancies (n=24) compared with CRC (p<0.0001). CONCLUSIONS: MPCT appears to have prognostic potential in this patient population, with significant reductions in survival if a CRC or ECA is detected. The detection of ECA would appear to have at least as important an impact on the usefulness of the examination as the detection of CRC.

Clinical characteristics of an anatomical hand index measured in patients with rheumatoid arthritis as a potential outcome measure.

OBJECTIVES: To determine the clinical characteristics of an anatomical hand index previously reported as a potential measure of joint deformity and outcome in patients with rheumatoid arthritis. METHODS: The hand index (open hand span - closed hand span/lateral height of the hand) was measured in a cross-sectional study of 145 out-patients with rheumatoid arthritis with disease durations 0-55 yr. Subsets of patients were restudied at mean follow-ups of approximately 9 months and 4 yr. RESULTS: The hand index fell gradually with disease duration. Correlations were demonstrated with the Sharp index (r = - 0.39, P = 0.000) and to a lesser extent with disease activity score (r = - 0.28, P = 0.001). At 260 +/- 115 days the hand index worsened by 0.09 units (P = 0.09, NS). At 51.6 +/- 5.4 months the index showed a fall from 1.96 +/- 0.73 to 1.61 +/- 0.65 (P = 0.000). During the same interval the Sharp index increased from 60 +/- 68 to 80 +/- 71 (P = 0.000). CONCLUSIONS: Measurement of simple hand dimensions can demonstrate worsening of hand deformity with time in patients with rheumatoid arthritis. We suggest that more sophisticated analysis of digital hand images, as used in our original study, might yield additional information and increase the sensitivity of an anatomical hand index as an outcome measure in rheumatoid arthritis.

The use of mammography in diagnosis and screening.

Up to nine per cent of women are destined to have breast cancer. Of these 25% are likely to be occult lesions only detectable on mammography. Early detection in important. Ten year survivals of over 65% can be expected in patients without axillary node involvement, compared to 25% with nodes. Apart from biopsy, mammography is the most sensitive method of cancer detection and is therefore of considerable help to the surgeon in evaluation of breast disease.

Flattened fundus sign of the septate gallbladder.

The fundus of a normal gallbladder is rounded. Any deviation from this configuration should prompt a search for the nonfilled segment of a septate gallbladder. Three illustrative cases are presented in which the initial observation of a flat fundus led to further evaluation and visualization of the remaining portion of a septate gallbladder.

Pointers to the diagnosis of gastric lymphoma on computed tomography.

The computed tomographic features of 12 patients with primary gastric non-Hodgkin's lymphoma and a further seven patients with gastric involvement by disseminated non-Hodgkin's lymphoma have been reviewed. In eight of these 19 patients there were clefts within the area of gastric wall involved by lymphoma where oral contrast medium passed to within 2 mm of the serosal surface in and amongst areas of gastric wall thickening (maximal thickness ranging from 18 to 40 mm). In three of these patients some clefts extended beyond the expected margin of the stomach to enter into, and be contained by, a composite soft tissue mass of stomach, spleen and splenic hilar nodes. Awareness of these features, which are in keeping with the known range of macroscopic pathological appearances in non-Hodgkin's lymphoma, may help the radiologist to recognize this potentially curable disease.