pkl1(+)and klp2(+): Two kinesins of the Kar3 subfamily in fission yeast perform different functions in both mitosis and meiosis.

We have identified Klp2p, a new kinesin-like protein (KLP) of the KAR3 subfamily in fission yeast. The motor domain of this protein is 61% identical and 71% similar to Pkl1p, another fission yeast KAR3 protein, yet the two enzymes are different in behavior and function. Pkl1p is nuclear throughout the cell cycle, whereas Klp2p is cytoplasmic during interphase. During mitosis Klp2p enters the nucleus where it forms about six chromatin-associated dots. In metaphase-arrested cells these migrate back and forth across the nucleus. During early anaphase they segregate with the chromosomes into two sets of about three, fade, and are replaced by other dots that form on the spindle interzone. Neither klp2(+) nor pkl1(+) is essential, and the double deletion is also wild type for both vegetative and sexual reproduction. Each deletion rescues different alleles of cut7(ts), a KLP that contributes to spindle formation and elongation. When either or both deletions are combined with a dynein deletion, vegetative growth is normal, but sexual reproduction fails: klp2 Delta,dhc1-d1 in karyogamy, pkl1 Delta,dhc1-d1 in multiple phases of meiosis, and the triple deletion in both. Deletion of Klp2p elongates a metaphase-arrested spindle, but pkl1 Delta shortens it. The anaphase spindle of klp2 Delta becomes longer than the cell, leading it to curl around the cell's ends. Apparently, Klp2p promotes spindle disassembly and contributes to the behavior of mitotic chromosomes.

Prospective study of routine perioperative transesophageal echocardiography for elective valve replacement: clinical impact and cost-saving implications.

Transesophageal echocardiography (TEE) is widely used during heart valve replacement operations, but its clinical impact and cost-saving profile have not been studied prospectively for this indication. We investigated the clinical benefits and cost-savings of routine TEE for elective valve replacement at a regional tertiary center. We prospectively studied 300 patients (140 men; mean age [+/-SD], 66 +/- 9 years) undergoing aortic valve, mitral valve, or double-valve replacements. Transesophageal echocardiography with a biplane (in 161 patients) or a multiplane probe was performed before and after surgery. We assessed whether the TEE findings changed the operation or the postoperative treatment and the cost of TEE either as an extension of a preexisting service or as a new development. In 2 patients undergoing aortic valve replacement, significant mitral regurgitation on TEE led to additional mitral valve replacement, and in 1 patient undergoing mitral valve replacement, aortic regurgitation also required aortic valve replacement. Immediate reoperation (dehisced mitral valve prosthesis) and delayed extubation (suspected obstruction of an aortic valve prosthesis) were prompted by postoperative TEE. Extending an existing TEE service to routine intraoperative use saved up to $109 (US) per patient per year. Routine intraoperative TEE can provide major clinical benefit to a small proportion of patients undergoing elective valve replacement, and this can lead to cost savings, but only if the service can be provided without major capital investment.

Oncogene mutation and prognosis in the myelodysplastic syndromes.

In a series of myelodysplastic syndrome patients, mutational status (particularly RAS mutation) was found to be prognostic for survival, independently of four previously reported scoring systems (Bournemouth, Sanz, Lille, International).

Identification of the lipophilic factor produced by macrophages that stimulates steroidogenesis.

Macrophages are known to release a lipophilic factor that stimulates testosterone production by Leydig cells. This macrophage-derived factor (MDF) is thought to be physiologically relevant, because removal of macrophages from the testis results in altered testosterone secretion and reduced fertility. The purpose of the present study was to purify this factor, elucidate its chemical structure, and determine whether it is both present in the testis and acts when injected intratesticularly. Culture media from testicular and peritoneal macrophages were extracted with ether, and the organic phase was sequentially purified on C18, silica, and cyano-HPLC columns. MDF was detected using a rat Leydig cell bioassay, with testosterone secretion being the end point. Purified material and crude ether extracts were analyzed by gas chromatography/mass spectrometry and nuclear magnetic resonance spectroscopy. The time of elution of MDF from both testicular and peritoneal macrophages was identical on all three HPLC columns. A single peak was observed when MDF, obtained from the final HPLC column, was analyzed by gas chromatography. The MS fragmentation pattern of purified material from both peritoneal and testicular macrophages was identical to that of a reference preparation of 25-hydroxycholesterol. Also, the nuclear magnetic resonance spectrum of MDF was similar to that of authentic 25-hydroxycholesterol. When 25-hydroxycholesterol was subjected to the identical purification scheme as MDF, it was found to elute at the same times as MDF on all three columns and elicited activity in the Leydig cell bioassay as expected. Control medium purified identically did not contain 25-hydroxycholesterol or have biological activity. Ether extracts of testis contained 25-hydroxycholesterol, indicating that this compound is present under physiological conditions. Similarly, when 25-hydroxycholesterol was injected into the testis of adult rats, testosterone production was increased within 3 h. Taken together, these data indicate that the lipophilic factor produced by macrophages that stimulates steroidogenesis is 25-hydroxycholesterol.

Cytogenetic abnormalities in the myelodysplastic syndromes and occupational or environmental exposure.

Patients with myelodysplastic syndromes (MDS) have high frequencies of cytogenetic abnormalities and evidence is accumulating of associations between exposure history and primary MDS. The objective of this article is to examine the relationship between histories of occupational or environmental exposure and presence of cytogenetic abnormalities. A case control study of MDS patients estimated lifetime exposure to more than 90 potential hazards in 400 age, sex, and area of residence matched patient and control pairs. A parallel cytogenetics study undertaken at time of diagnosis, independently of any knowledge of exposure history, identified 75 cytogenetically abnormal and 139 normal (186 not studied). Odds ratios of MDS patients and their matched controls were compared for 3 groups: cytogenetically abnormal, normal, and not known. The odds ratios for all exposures combined were possibly higher among cytogenetically abnormal 2.0 (95% confidence interval 0.8-5.9) than among normal 1.0 (0.6-1.8). This pattern was observed for exposure to semimetals, abnormal 4.0 (0.4-195.1) and normal 0.5 (0.1-1.0) and inorganic dusts, 1.6 (0. 6-3.8) and 0.4 (0.1-1.4) respectively. The pattern was principally in abnormalities in chromosomes 5 and 7. For organic chemicals and radiation, the odds ratios for both cytogenetically abnormal and normal were marginally raised: organic 1.8 (0.6-6.0) and 1.3 (0.6-2.9), respectively, and radiation 1.7 (0.5-5.6) and 1.3 (0.4-4.7) respectively. For radiation, abnormalities were mostly in chromosome 8. This study of association between exposures and cytogenetics in primary MDS complements those previously reported in secondary MDS and may provide some insight into pathogenetic mechanisms that lead to development of MDS. (Blood. 2000;95:2093-2097)

A cytoplasmic dynein heavy chain is required for oscillatory nuclear movement of meiotic prophase and efficient meiotic recombination in fission yeast.

Meiotic recombination requires pairing of homologous chromosomes, the mechanisms of which remain largely unknown. When pairing occurs during meiotic prophase in fission yeast, the nucleus oscillates between the cell poles driven by astral microtubules. During these oscillations, the telomeres are clustered at the spindle pole body (SPB), located at the leading edge of the moving nucleus and the rest of each chromosome dangles behind. Here, we show that the oscillatory nuclear movement of meiotic prophase is dependent on cytoplasmic dynein. We have cloned the gene encoding a cytoplasmic dynein heavy chain of fission yeast. Most of the cells disrupted for the gene show no gross defect during mitosis and complete meiosis to form four viable spores, but they lack the nuclear movements of meiotic prophase. Thus, the dynein heavy chain is required for these oscillatory movements. Consistent with its essential role in such nuclear movement, dynein heavy chain tagged with green fluorescent protein (GFP) is localized at astral microtubules and the SPB during the movements. In dynein-disrupted cells, meiotic recombination is significantly reduced, indicating that the dynein function is also required for efficient meiotic recombination. In accordance with the reduced recombination, which leads to reduced crossing over, chromosome missegregation is increased in the mutant. Moreover, both the formation of a single cluster of centromeres and the colocalization of homologous regions on a pair of homologous chromosomes are significantly inhibited in the mutant. These results strongly suggest that the dynein-driven nuclear movements of meiotic prophase are necessary for efficient pairing of homologous chromosomes in fission yeast, which in turn promotes efficient meiotic recombination.

Occupational exposures and haematological abnormalities among ordnance factory workers: a case control study.

The lifetime exposures to an extensive list of chemical and other potentially toxic hazards were estimated for all available employees at an ordnance factory by questionnaire-based interview. Exposure histories of 32 (of 33) previously diagnosed as haematologically abnormal (cases) were compared with 322 (of 345) normals (controls). Among 'ordnance factory chemicals', modestly increased odds ratios were observed for men for acetic anhydride (2.8), stearic acid (2.8), and possibly for resorcinol (2.9), TNT (2.4) and hydroxy terminated polybutadene (HTPB) (2.4). Increased odds ratios were also observed in exposures not directly related to ordnance manufacture, including mineral acids, welding fumes, exhaust gases and insecticides. While, small numbers in the case group limit the statistical significance of reported odds ratios, one should be cautious about committing a type II error. These findings may partly explain the previously reported unusual prevalence of haematological abnormalities within the factory.

Psychological rehabilitation after myocardial infarction: multicentre randomised controlled trial.

OBJECTIVE: To evaluate rehabilitation after myocardial infarction. DESIGN: Randomised controlled trial of rehabilitation in unselected myocardial infarction patients in six centres, baseline data being collected on admission and by structured interview (of patients and spouses) shortly after discharge and outcome being assessed by structured interview at six months and clinical examination at 12 months. SETTING: Six district general hospitals. SUBJECTS: All 2328 eligible patients admitted over two years with confirmed myocardial infarction and discharged home within 28 days. INTERVENTIONS: Rehabilitation programmes comprising psychological therapy, counselling, relaxation training, and stress management training over seven weekly group outpatient sessions for patients and spouses. MAIN OUTCOME MEASURES: Anxiety, depression, quality of life, morbidity, use of medication, and mortality. RESULTS: At six months there were no significant differences between rehabilitation patients and controls in reported anxiety (prevalence 33%) or depression (19%). Rehabilitation patients reported a lower frequency of angina (median three versus four episodes a week), medication, and physical activity. At 12 months there were no differences in clinical complications, clinical sequelae, or mortality. CONCLUSIONS: Rehabilitation programmes based on psychological therapy, counselling, relaxation training, and stress management seem to offer little objective benefit to patients who have experienced myocardial infarction compared with previous reports of smaller trials.

Occupational and environmental exposures and myelodysplasia: a case-control study.

A case-control study of newly diagnosed myelodysplastic syndrome patients investigated lifetime exposures through occupation, environment or hobby by questionnaire, structured and semi-structured interview. The exposure histories of 400 individually matched pairs were compared. Increased or possibly increased odds ratios were observed for radiation (2.05, 95% confidence interval 1.16-3.76), halogenated organics (1.57, 0.97-2.57), metals (1.40, 0.99-2.00), several specific radiation exposures and individual chemicals and for childlessness (1.46, 1.01-2.11). Since myelodysplasia generally carries a poor prognosis, whether or not individuals convert to leukaemia or to other cancer, these findings add to previous reports of exposures implicated in the aetiology of leukaemia and add to the case for minimizing exposures to radiation and halogenated organics.

Topoisomerase II alpha expression in acute myeloid leukaemia and its relationship to clinical outcome.

The treatment of acute myeloid leukaemia (AML) is often complicated by resistance to chemotherapeutic drugs. Many of the most effective drugs used to treat haematological malignancies target the nuclear enzyme topoisomerase II. Resistance to these drugs in vitro has been associated with quantitative and qualitative changes in the enzyme. In this study, we have investigated topoisomerase II mRNA expression in leukaemic blasts from 23 AML patients. Expression levels ranged from 1-47% relative to the haemopoietic cell line HL60 in 16 evaluable patients. Thirteen of 16 patients achieved complete remission (CR). We have therefore chosen ease of entry into CR as the most sensitive clinical correlate. Decreased topoisomerase II mRNA expression in vitro results in drug resistance. The clinical relevance of reduced expression is not known. All cases of AML and de novo AML have been studied separately. We are unable to identify a correlation between topoisomerase II mRNA levels and ease of entry into CR in either of these groups. Taking these findings into account, group size calculations have been undertaken and indicate that a multi-centre study of this question is warranted.

Effect of systemic vasodilators on internal mammary artery flow. Implications for postoperative treatment after myocardial revascularization.

Postoperative spasm of the internal mammary artery graft can cause morbidity and mortality after myocardial revascularization. To our knowledge, the ability of systemic vasodilators to overcome internal mammary artery spasm has not been studied clinically. In 50 patients in whom the left internal mammary artery was used for myocardial revascularization, we have investigated the effect of five agents on internal mammary artery free flow: normal saline, dobutamine, glyceryl trinitrate, sodium nitroprusside, and enoximone, a phosphodiesterase III inhibitor. After the internal mammary artery was harvested, free flow was measured under controlled hemodynamic conditions before any pharmacologic intervention (flow 1) and a mean of 18.5 +/- 3 (standard deviation) minutes after a systemic infusion of one of the five agents was begun (flow 2). The increase in free flow expressed as a percentage of initial flow was greater for enoximone (94% +/- 24%) than for normal saline (18% +/- 11%), dobutamine (40% +/- 27%), and glyceryl trinitrate (52% +/- 36%) (all three p < 0.01). The increase in flow for sodium nitroprusside (78% +/- 37%) was greater than for normal saline and dobutamine (both p < 0.05). We therefore recommend the systemic use of enoximone and sodium nitroprusside, in rank order, to prevent and treat postoperative spasm of the internal mammary artery.

Coronary artery bypass surgery: current practice in the United Kingdom.

OBJECTIVE: To assess current clinical practice in coronary artery bypass surgery and compare it with a previous survey conducted five years ago. SETTING: United Kingdom. DESIGN: Postal questionnaires were sent in March 1993 to 120 consultant cardiac surgeons currently performing coronary artery bypass surgery. 104 (87%) were returned by May 1993. RESULTS: The 104 surgeons who returned the questionnaire performed an estimated total of 25,234 coronary artery bypass operations in 1992 with an average case load per surgeon similar to that in 1987 (243 v 214, NS). The internal mammary artery was regarded as the conduit of choice by 101 surgeons (97%) and was used in 93% of bypass grafts to the left anterior descending coronary artery compared with 73% in 1987 (p < 0.001) but only in 7% of grafts to the circumflex and right coronary systems. There was also a significant increase in the number of surgeons using both internal mammary arteries (88% v 59%, p < 0.01) but only a small increase in those using the internal mammary artery as a sequential graft (55% v 44%, NS). The age of the patient remains one of the main contraindications to the use of the internal mammary artery (40%), together with insufficient mammary flow (42%), endarterectomy (22%), and unstable angina (17%). The right gastroepiploic and inferior epigastric arteries were used only occasionally (3%) when the internal mammary artery or the saphenous vein were not available. Most surgeons (96%) still advocate the use of aspirin to enhance graft patency, with 87% of surgeons continuing treatment indefinitely, compared with 50% in the previous survey (p < 0.001). As for methods of myocardial protection, 72% of surgeons used cardioplegic arrest whereas 28% preferred intermittent aortic cross clamping and fibrillation. CONCLUSIONS: It is the consensus among British cardiac surgeons that the internal mammary artery is the graft conduit of choice. Its use has been significantly extended over the past five years (1987 to 1992) suggesting a quick response to advancing scientific knowledge. The use of alternative arterial conduits is still limited, perhaps as a reflection of the relative lack of information on their long-term performance. The recently advocated technique of retrograde cardioplegia and continuous warm cardioplegia is not yet popular.

The effect of MDR-1 gene expression on outcome in acute myeloblastic leukaemia.

Resistance to cytotoxic agents may be encountered during the treatment of acute myeloblastic leukaemia (AML). P-glycoprotein encoded by the MDR-1 gene has been implicated as a potential drug resistance mechanism in leukaemic cells. In recent years, many data have been accrued concerning the expression of P-glycoprotein in leukaemia, and several studies have been published which have related MDR status to outcome in AML. Conclusions as to the effect of P-glycoprotein expression on prognosis in AML have varied widely. The studies are not immediately comparable, since they differ in methodology, treatment regimens, demographic profile and, perhaps most importantly, criteria for positivity of MDR status. The technique of statistical overview (meta-analysis) can be used to pool observational studies. Application of this statistical method to existing studies suggests an estimated relative risk of 0.68 for P-glycoprotein expression with respect to complete remission in AML. Further large studies are required to determine fully the role of P-glycoprotein in AML.

Median sternotomy wound dehiscence: a retrospective case control study of risk factors and outcome.

Over a 5-year period from 1985 to 1989, 2760 patients underwent open heart surgery at the University Hospital of Wales. Of these, 44 (1.6%, 35 men, mean age 61 years) developed median sternotomy dehiscence 2-40 (median 9) days after surgery. Infection was an associated factor in 18 patients (41%) and Staphylococcus aureus the predominant isolate in seven of those. Thirty-seven patients underwent rewiring of the sternotomy wound and seven patients underwent debridement, removal of wires and delayed closure. In those undergoing rewiring, sternal stability was maintained in 34 patients (92%). There were seven deaths (16%), of which two were considered to be wound-related. Median hospital stay of survivors was 34 (range 16-84) days. Comparison with 88 matched controls by univariate analysis showed preoperative chronic obstructive airways disease, reduced FEV1/FVC ratio (both P < 0.001) and smoking (P < 0.05) were all more common in the dehiscence group. In addition, reoperation for bleeding (P < 0.05), prolonged bypass time, postoperative ventilation period and length of stay in the intensive care unit (all P < 0.001) were more common in the study group.

Smoking: its influence on survival and cause of death.

Smoking is strongly associated with age-specific death rates for a number of diseases. Increased age-specific death rates for a disease may imply either more deaths from the disease with increased absolute lifetime risk, or earlier deaths, without increased absolute lifetime risk. The British doctor smoking data are re-analysed using lifetable methods for survival, cumulative mortality and the disease-specific cumulative mortality. The most significant effect of smoking is on survival: life shortening amounts to three years for light smokers, five for moderate smokers, and eight for heavy smokers, compared with those who never smoked. Smoking increases the absolute number of deaths from some causes, including lung cancer; for other causes, including ischaemic heart disease, it brings forward death without increasing the absolute number of deaths. The smoking associations with more or earlier death have implications for research into the mechanisms of disease causation, for health promotion, for rational health-care planning, and for social policy.

A case control study of factors associated with macroscopic breast cysts.

Women who have a breast cyst aspirated are at increased risk of developing breast cancer. We present an age-matched case control study comparing the reproductive characteristics of 352 women who had a breast cyst aspirated with a control group of 352 contemporaneous clinically normal women. Women with breast cysts were more likely to be nulliparous [odds ratio (OR) = 2.28, 95% confidence interval (CI) = 1.34-3.88] or have a late age at first live birth (chi 2 trend = 5.6, P less than 0.025), and a late menopause (chi 2 trend = 4.3, P less than 0.05). They were less likely to have ever used the oral contraceptive pill (OR = 0.38, 95% CI = 0.26-0.55) or to have used the pill for a short duration (chi 2 trend = 16.8, P less than 0.001), and were less likely to have had a hysterectomy (OR = 0.58, 95% CI = 0.36-0.93). They were more likely to wear a small bra (chi 2 trend = 18.6, P less than 0.001) and bra cup (chi 2 trend = 5.6, P less than 0.025). Nulliparity, late age at first live birth and late menopause are factors common to breast cancer risk.

Reproductive factors associated with mastalgia.

Mastalgia has been classified as an aberration of the normal, rather than a disease. We present an age-matched, case-control study which compares the reproductive factors of patients with severe cyclical and noncyclical mastalgia with control groups of women who presented to a breast clinic but in whom no clinical abnormality was found and who did not later develop breast disease. There were few statistically significant differences in reproductive factors between the cases and controls, which lends some support to the concept that mastalgia can be considered to be an aberration of the normal.

Specificity of association in analysis of mortality and inference on causality.

Specificity of association between putative risk factor and disease under study is important to inference on causality. Nevertheless many studies investigate mortality of a single disease without comparison with a control. Age-standardized proportional mortality ratios make single disease studies into case-control studies and thus demonstrate whether or not associations are disease specific. Comparison of disease-specific with all-cause mortality experiences of whole populations classified by exposure, clearly distinguishes between exposures associated with more death and with earlier/younger death, thereby overcoming an important limitation of the familiar standardized mortality ratio (SMR). Smoking is associated with more death from lung cancer (lifetime cause-specific proportions, never 1%, light 6%, moderate 8% and heavy 12%) and with earlier/younger death from ischaemic heart disease (never 35%, light 34%, moderate 32% and heavy 29%).

Is there an increased risk of breast cancer in women who have had a breast cyst aspirated?

A consecutive series of 644 women who presented with breast nodularity between 1976 and 1982 have been followed up to determine their rate of subsequent breast cancer. Fifteen women have developed breast cancer, 14 of these were among 352 women with an aspirated cyst (relative risk 4.4). Women with multiple cysts had the highest risk and women with breast nodularity had no excess risk. Review of histology specimens from those women who had undergone biopsy showed an excess of florid epithelial hyperplasia in women who subsequently developed breast cancer and women with multiple aspirated cysts were more likely to have florid epithelial hyperplasia. Multiple cysts are clinical markers of histological breast proliferation and women who have had multiple breast cysts aspirated have an increased risk of breast cancer and should be advised to practice regular self examination.