Characterization of the I4399M variant of apolipoprotein(a): implications for altered prothrombotic properties of lipoprotein(a).

Essentials Elevated lipoproteinp(a) is an independent and causal risk factor for atherothrombotic diseases. rs3798220 (Ile/Met substitution in apo(a) protease-like domain) is associated with disease risk. Recombinant I4399M apo(a) altered clot structure to accelerate coagulation/delay fibrinolysis. Evidence was found for increased solvent exposure and oxidation of Met residue. SUMMARY: Background Lipoprotein(a) (Lp[a]) is a causal risk factor for a variety of cardiovascular diseases. Apolipoprotein(a) (apo[a]), the distinguishing component of Lp(a), is homologous with plasminogen, suggesting that Lp(a) can interfere with the normal fibrinolytic functions of plasminogen. This has implications for the persistence of fibrin clots in the vasculature and hence for atherothrombotic diseases. A single-nucleotide polymorphism (SNP) (rs3798220) in the gene encoding apo(a) has been reported that results in an Ile→Met substitution in the protease-like domain (I4399M variant). In population studies, the I4399M variant has been correlated with elevated plasma Lp(a) levels and higher coronary heart disease risk, and carriers of the SNP had increased cardiovascular benefit from aspirin therapy. In vitro studies suggested an antifibrinolytic role for Lp(a) containing this variant. Objectives We performed a series of experiments to assess the effect of the Ile→Met substitution on fibrin clot formation and lysis, and on the architecture of the clots. Results We found that the Met variant decreased coagulation time and increased fibrin clot lysis time as compared with wild-type apo(a). Furthermore, we observed that the presence of the Met variant significantly increased fibrin fiber width in plasma clots formed ex vivo, while having no effect on fiber density. Mass spectrometry analysis of a recombinant apo(a) species containing the Met variant revealed sulfoxide modification of the Met residue. Conclusions Our data suggest that the I4399M variant differs structurally from wild-type apo(a), which may underlie key differences related to its effects on fibrin clot architecture and fibrinolysis.

Consequences of mating and predation risk for longevity in a freshwater snail: abstinence makes the heart beat longer.

Senescence is not a static property of an individual or population, but rather it is a dynamic process that may be influenced by environmental conditions. This can occur in at least two ways: in the long-term across multiple generations, and in the short-term via phenotypic plasticity. The former has attracted a lot of attention, both theoretically and empirically; the latter has lagged behind. To determine whether two important environmental variables (predation risk and mate availability) affect the pattern of actuarial senescence (i.e. the increase in mortality with age), we reared 30 full-sib families of the simultaneously hermaphroditic freshwater snail Physa acuta under four different environmental conditions and tracked individuals until death. Individuals were reared in a 2×2 factorial experiment that manipulated the nonlethal presence of chemical cues from predatory crayfish (presence/absence) and the opportunity to mate with an unrelated partner (mated/not mated). Snails that receive a partner reproduce by outcrossing, whereas those that remain in isolation can reproduce by self-fertilization. We compared the cumulative survival curves to test for an effect of predation risk and mating. The hazard ratio (HR) for the predation risk comparison was 1.042 indicating no significant difference between the curves. However, the HR for the mating comparison was 4.021, reflecting a significant reduction in survival probability for mated snails relative to isolated snails. As such, mating resulted in a much shorter lifespan, an outcome that we interpret in terms of shifting resource allocation.

Constraints on the evolution of phenotypic plasticity: limits and costs of phenotype and plasticity.

Phenotypic plasticity is ubiquitous and generally regarded as a key mechanism for enabling organisms to survive in the face of environmental change. Because no organism is infinitely or ideally plastic, theory suggests that there must be limits (for example, the lack of ability to produce an optimal trait) to the evolution of phenotypic plasticity, or that plasticity may have inherent significant costs. Yet numerous experimental studies have not detected widespread costs. Explicitly differentiating plasticity costs from phenotype costs, we re-evaluate fundamental questions of the limits to the evolution of plasticity and of generalists vs specialists. We advocate for the view that relaxed selection and variable selection intensities are likely more important constraints to the evolution of plasticity than the costs of plasticity. Some forms of plasticity, such as learning, may be inherently costly. In addition, we examine opportunities to offset costs of phenotypes through ontogeny, amelioration of phenotypic costs across environments, and the condition-dependent hypothesis. We propose avenues of further inquiry in the limits of plasticity using new and classic methods of ecological parameterization, phylogenetics and omics in the context of answering questions on the constraints of plasticity. Given plasticity's key role in coping with environmental change, approaches spanning the spectrum from applied to basic will greatly enrich our understanding of the evolution of plasticity and resolve our understanding of limits.

Evaluating the contributions of change in investment and change in efficiency to age-related declines in male and female reproduction.

It is commonly observed that reproduction decreases with age, often at a different rate in males and females. This phenomenon is generally interpreted as senescence. Such reproductive declines may stem from at least two sources: a change in resource allocation and a decline in the ability to convert resources into offspring. This distinction is important because a shift in resource allocation may be favoured by selection, while reduced efficiency is purely deleterious. We propose a way to distinguish whether a decline in reproduction is purely deleterious based on estimating reproductive investment, output, and their ratio, efficiency. We apply this approach to the hermaphroditic snail Physa acuta and demonstrate that both male and female functions decline with age. The male decline largely stems from reduced investment into male activity while female decline is due to increased reproductive inefficiency. This shows that age-related declines in reproduction can occur for a number of different reasons, a distinction that is usually masked by the general term 'senescence'. This approach could be applied to any species to evaluate age-related reproductive decline. We advocate that future studies measure age trajectories of reproductive investment and output to explore the potential processes hidden behind the observation that reproduction declines with age.

How useful is templating for total knee replacement component sizing?

This study aims to assess the accuracy of digital templating at our institution, by comparing the templated component sizes with those implanted, and to determine whether templating the preoperative films had any measurable difference on the radiographic outcome, and if, where there was a mismatch between the implanted and templated sizes, the templated size would have been preferable. While a number of studies have evaluated the accuracy of both acetate and digital templating, none has to our knowledge looked back at post-operative radiographs and reviewed these in light of the templated and implanted sizes. Data was collected from 90 PFC Sigma (DePuy, UK) total knee replacements done sequentially, 45 of whom were templated digitally using a calibrating ball and Agfa Orthopaedic Tools software. Postoperative radiographs were graded independently for correct sizing. All templates were within one size of the implanted prosthesis. The femoral component appeared to be more often oversized on the postoperative radiographs in the non-templated group. In addition, most tibial trays that were found be too small had been templated to a larger size. There was a trend towards tibial trays templated too large to have been templated to a smaller size. We conclude that digital templating with a calibrating device is a useful part of preoperative planning for total knee arthroplasty.

Are there interactive effects of mate availability and predation risk on life history and defence in a simultaneous hermaphrodite?

Encountering mates and avoiding predators are ubiquitous challenges faced by many organisms and they can affect the expression of many traits including growth, timing of maturity and resource allocation to reproduction. However, these two factors are commonly considered in isolation rather than simultaneously. We examined whether predation risk and mate availability interact to affect morphology and life-history traits (including lifetime fecundity) of a hermaphroditic snail (Physa acuta). We found that mate availability reduced juvenile growth rate and final size. Predator cues from crayfish induced delayed reproduction, but there were no reduced fecundity costs associated with predator induction. Although there were interactive effects on longevity, lifetime fecundity was determined by the number of reproductive days. Therefore, our results indicate a resource-allocation trade-off among growth, longevity and reproduction. Future consideration of this interaction will be important for understanding how resource-allocation plasticity affects the integration of defensive, life-history and mating-system traits.

Implant-related fractures of the femur following hip fracture surgery.

BACKGROUND: Most hip fractures are treated surgically, with use of either internal fixation or prosthetic replacement of the femoral head. The presence of these implants increases the risk of a later femoral fracture in susceptible osteoporotic patients. The purpose of this study was to analyze the incidence of and risk factors for implant-related fractures of the femur after previous hip fracture surgery. METHODS: Over a ten-year period from January 1988 to December 1997, 6230 patients (median age, eighty-two years; male:female ratio, 1247:4983) who sustained a total of 6696 hip fractures were admitted to the Edinburgh Orthopaedic Trauma Unit. Demographic information on the patients and details of the original treatment of the hip fracture were prospectively coded and entered into a trauma database. All subsequent readmissions due to a femoral fracture related to the implant were prospectively audited and extracted for the purposes of this study. RESULTS: One hundred and forty-one patients sustained an ipsilateral fracture of the femur at a median of twenty-four weeks following the original hip fracture surgery. Survivorship analysis of the hip fracture population revealed an overall rate of subsequent femoral fracture of 2.9% at five years, which increased to 5.1% at ten years. The median age and gender distribution of the patients who sustained a subsequent femoral fracture were similar to those of the hip fracture population as a whole. Two-thirds of the fractures propagated from the tip of the implant. Analysis of the subsequent fractures according to the type of implant used to treat the original fracture revealed considerable differences in incidence. The incidence was relatively high in the patients initially treated with a Gamma nail (18.74 fractures per 1000 person-years) or a cementless hemiarthroplasty (11.72 per 1000 person-years) and was relatively low in those treated with a compression hip screw (4.46 per 1000 person-years), cannulated screws (4.50 per 1000 person-years), or a primary arthroplasty with cement (6.2 per 1000 person-years). The highest incidence of fracture was seen in the patients who had required an arthroplasty with cement as a revision procedure following failure of a primary implant (22.39 per 1000 person-years). CONCLUSIONS: Implant-related fractures following hip fracture surgery are more common than has previously been appreciated. The risk of later ipsilateral femoral fracture is increased by the use of a Gamma nail or a cementless hemiarthroplasty to treat the original hip fracture.

Possible role of endothelin-1 in the rabbit urinary bladder hyperplasia secondary to partial bladder outlet obstruction.

OBJECTIVES: Urinary bladder hypertrophy and hyperplasia are common features of bladder outlet obstruction (BOO). The urinary bladder is known to synthesize endothelin-1 (ET-1), which is a potent vasoconstrictor peptide with mitogenic properties. Using an animal model of partial BOO, we investigated the potential role of ET-1 and its receptor subtypes (ET(A) and ET(B)) in bladder smooth muscle cell (SMC) proliferation. MATERIALS AND METHODS: Partial BOO was produced in adult male New Zealand White rabbits. After 3 weeks, the bladder was removed and SMCs from the dome and bladder neck were grown using standard explant methodology. At passage 2, the cells were made quiescent and then further incubated in foetal calf serum (FCS), control age-matched rabbit serum (CRS) or partial BOO serum (BRS) in the presence or absence of ET(A)-antagonist (BQ123) or ET(B)-antagonist (BQ788). SMC proliferation was then measured 24 h later with 5-bromo-2'deoxy-uracil and by cell counting using a haemocytometer at 48 h. Immunostaining for alpha-actin was performed on detrusor and bladder neck cells to confirm the presence of smooth muscle cells. RESULTS: BQ123 and BQ788 did not influence detrusor or bladder neck SMC proliferation in FCS or CRS. However, in the presence of BRS, BQ123 and BQ788 (100 nmol/L) significantly (p = 0.008) inhibited detrusor and bladder neck SMC proliferation. Cell counts were significantly reduced from the detrusor (p = 0.03, p = 0.01 with BQ123 and BQ788, respectively) and bladder neck (p = 0.01 for both BQ123 and BQ78). CONCLUSIONS: These results suggest that ET antagonists may have a role in preventing SMC hyperplasia associated with partial BOO.

Down-regulation of endothelin-B receptor sites in cavernosal tissue of a rabbit model of partial bladder outlet obstruction: potential clinical relevance.

Erectile dysfunction (ED) is a common problem that significantly affects quality of life and psychological well-being. Benign prostatic hyperplasia (BPH) is the commonest known benign proliferative disorder. Recently there has been growing evidence to suggest that patients with high BPH symptom scores have an increased incidence of ED. Endothelin-1 (ET-1) is a potent vasoconstrictor peptide that is thought to play an important role as a modulator of erectile physiology and dysfunction. We investigated whether there were any changes in the penile histology and in the density and distribution of ET-1 and its receptor subtypes in the corpora cavernosa of a rabbit model of partial bladder outflow obstruction (BOO). BOO was induced in 12 adult New Zealand White rabbits; 12 sham-operated rabbits acted as controls. Penises were excised after 3 and 6 weeks (n=6 each for control and BOO). Low- and high-resolution autoradiography was performed using radioligands for ET-1, ET(A) and ET(B) receptors and the results were analysed densitometrically. Ultrastructural evaluation of the corpus cavernosum (CC) was also performed. ET-1, ET(A) and ET(B) receptor-binding sites were primarily localised to the smooth-muscle cells (SMC) of the CC and to the endothelium lining the cavernosal space. ET(B) receptor-binding sites were significantly decreased (P=0.04) in the 6-week BOO cavernosal tissue. These receptor changes were accompanied by ultrastructural changes in the CC. ET-1 may play a role in the pathophysiology of ED associated with BPH. This may partly be due to enhanced vasoconstrictor actions and SMC proliferation secondary to a reduction in ET(B) receptors. Further work is needed to evaluate this possibility.

An immunocytochemical study of MHC class I expression on human Langerhans cells and melanocytes.

Classical MHC class I glycoproteins (HLA-A, B, and C) present endogenous cytosolic peptide antigen fragments to CD8-positive T-cells. CD8-positive T-cell recognition and destruction of virus-infected cells are dependent on adequate cellular MHC class I expression. Constitutive MHC class I expression is ubiquitous, but known to be deficient on specific differentiated cell types which include hepatocytes, neurones, chondrocytes and myocytes. Although enabling assessment of MHC class I expression on individual cells, limitations of immunocytochemistry were encountered with this assessment on Langerhans cells and melanocytes. These dispersed intraepidermal cells were obscured by adjacent keratinocytes in sections immunostained for MHC class I glycoproteins. Initiatives designed to resolve the issue have included immunoelectron microscopy, cell culture techniques, and animal bone marrow chimera models. Despite the elegance of these techniques, the issue of MHC class I expression on Langerhans cells and melanocytes remains unresolved. In this immunocytochemical study, an alternative strategy was based upon the recognized deficiency of epithelial MHC class I expression within pilosebaceous adnexal units. Langerhans cells and melanocytes were therefore studied within this microenvironment of deficient MHC class I expression, using monomorphic and polymorphic MHC markers. Langerhans cells and melanocytes were demonstrated within pilosebaceous units of scalp skin by immunocytochemistry. Differentiation markers OKT6 (CD1a) and TMH1 defined Langerhans cells and melanocytes, respectively. Monomorphic MHC markers W6/32 and TAL IB5 defined invariant epitopes of HLA class I and II, respectively. Polymorphic MHC class I markers defined the HLA-Bw4 and HLA-Bw6 supertypic determinants. Constitutive MHC class I expression was shown to be deficient on Langerhans cells and melanocytes.

Descriptive epidemiology of primary cancer of the brain, cranial nerves, and cranial meninges in New Zealand, 1948-88.

We used New Zealand data on occurrence of different types of brain cancer to investigate: (i) a possible secular increase which has been seen worldwide and has generated considerable debate; (ii) possibly higher rates among Maori; and (iii) possibly higher risks related to social class and occupation. Data from the NZ Cancer Registry on the 5,684 brain cancers diagnosed among NZ residents from 1948-88 were used to study the pattern of occurrence by gender, age, race, calendar year, social class, occupation, and histology. Age-standardized brain-cancer incidence rates per 100,000 more than doubled over the 41-year period (from 2.9 to 6.9 in males and from 2.1 to 5.1 in females). A strong trend of increasing incidence with increasing social class is seen in males (Ptrend = 0.01). Among Maori, the proportion of all brain cancer that is medulloblastoma is four times that among non-Maori, and the proportion of all brain cancers that lack histologic confirmation is about 40 percent higher. Elevated risks are seen among: dairy farmers (odds ratio [OR] = 3.4, 95 percent confidence interval [CI] = 1.9-6.0); sheep handlers (OR = 2.7, CI = 1.4-5.3); livestock workers (OR = 3.8, CI = 1.7-8.4); and farm managers (OR = 3.2, CI = 1.4-7.2); as well as among electrical engineers (OR = 8.2, CI = 20-34.7); electricians (OR = 4.6, CI = 1.7-12.2); and other electrical workers. Brain cancer rates in NZ have increased steadily since 1948, but this increase has leveled off in the most recent five-year period. Although brain cancer rates are likely to be underestimated among the Maori, an excess of medulloblastoma is evident in this group.

Geographical distribution of cancers of the kidney and urinary tract and analgesic nephropathy in Australia and New Zealand.

Age-standardised incidence rates for cancers of the renal parenchyma, renal pelvis and bladder and for end-stage renal failure due to analgesic nephropathy for the years 1982-83 were compared between the Australian states and New Zealand, and within New South Wales (NSW), to determine whether these rates paralleled the previous prevalence of consumption of phenacetin-containing analgesics. Whereas little variation was seen within Australasia in respect of the incidence of cancer of the renal parenchyma and bladder, both cancer of the renal pelvis and end-stage renal failure due to analgesic nephropathy had higher incidence rates amongst women in NSW and Queensland than in the other states or New Zealand. Within NSW, the average annual incidence rates during 1973-82 for renal pelvic cancer in the Hunter region of 1.3 (m) and 1.6 (f) per 100,000 were the highest in the state. These high incidence rates coincided with areas known to have had a high prevalence of consumption of compound analgesics containing phenacetin. In an international comparison with populations which had published incidence rates for each of the periods 1973-77 and 1978-82, the rate for cancer of the renal pelvis in women was highest in both time periods in NSW and had increased absolutely at a faster rate.

Follow up of New Zealand participants in British atmospheric nuclear weapons tests in the Pacific.

OBJECTIVE: To study the health of Royal New Zealand Navy personnel who participated in atmospheric nuclear weapons tests conducted by the United Kingdom at Malden Island and Christmas Island in 1957 and 1958. DESIGN: Blinded, controlled follow up of up to 30 years. SETTING: New Zealand. SUBJECTS: 528 Men known to have participated in the tests and a control group of 1504 men who were in the Royal New Zealand Navy during the same period but did not participate in the tests. MAIN OUTCOME MEASURES: Mortality and incidence of cancer. RESULTS: Follow up for the period 1957-87 was 94% complete in test participants and 91% complete in the controls. There were 70 deaths among test participants and 179 deaths among controls, yielding a relative risk of 1.08 (90% confidence interval 0.85 to 1.38, p = 0.29). The relative risk of death from causes other than cancer was 0.96 (0.71 to 1.29, p = 0.59) whereas the relative risk of death from cancer was 1.38 (0.90 to 2.10, p = 0.09) and of the incidence of cancer was 1.12 (0.78 to 1.60, p = 0.29). For cancers other than haematological malignancies the relative risk was 1.14 (0.69 to 1.83, p = 0.31) for mortality and 1.01 (0.67 to 1.50, p = 0.48) for incidence. There were seven deaths from haematological cancers among test participants (relative risk 3.25, 90% confidence interval 1.12 to 9.64, p = 0.02), including four leukaemias (5.58, 1.04 to 41.6, p = 0.03). The relative risk for incidence of haematological cancers was 1.94 (0.74 to 4.84, p = 0.10) and that for leukaemia was 5.51 (1.03 to 41.1, p = 0.03). There were no cases of multiple myeloma in the test participants during the follow up period, but the expected number was only 0.3. CONCLUSIONS: Although the numbers are small, the findings for leukaemia are similar to those for British participants in the nuclear weapons test programme. Some leukaemias, and possibly some other haematological cancers, may have resulted from participation in this programme. There is little evidence of an increased risk for non-haematological cancers, and there is no evidence of an increased risk for causes of death other than cancer.