The Compress® transcutaneous implant for rehabilitation following limb amputation.

Amputation is an unfortunate outcome of a variety of orthopedic conditions. Many amputees can be functionally fitted with conventional suspension sockets. A substantial subset, however, fails this conventional treatment and is unable to function. In Europe, an alternative to socket-based prostheses has been available for 25 years. Patients there who are unable to functionally use socket-based prostheses have been offered the possibility for transcutaneous osseointegration. With this technology, the prosthetic limb can be rigidly attached to the residual bone, and the socket is eliminated, in many cases enabling improved function and patient satisfaction. In the United States, regulatory barriers have greatly limited the adoption and acceptance of transdermal osseointegration. The Compress® device was developed as an alternate means of fixation for massive endoprostheses, such as distal femoral replacements. A uniquely designed prosthesis is rigidly anchored to the end of the cortical bone and is then subjected to a large axial stress. The bone then grows avidly into the device, providing permanent osseointegration. We have recently adopted this device for transcutaneous use. These procedures have been performed in the United States on a custom regulatory basis. Results of this have been encouraging, and we are planning to begin a regulatory trial in the near future.

Regional variation in the incidence of abdominal aortic aneurysm in Sweden.

BACKGROUND: The risk factor profile is similar between patients with abdominal aortic aneurysm (AAA) and coronary heart disease (CHD). CHD is more common in the north of Sweden. It is unknown whether similar regional differences in the incidence of AAA exist. The aims of this study were to investigate whether there is a regional gradient of AAA incidence, and to compare time trends and the frequency of interventions between regions. METHODS: Swedish citizens have a 12-digit personal identification number. The Swedish Hospital Discharge Register covers inpatient care (diagnosis, admission, procedure codes, sex, date of birth, county). Population size was obtained from the central statistical bureau. Regions were south, mid and north. RESULTS: All records for 1990-2005 were extracted and 35 418 individuals with AAA were identified (74.8 per cent men). The highest age-standardized incidence (102.7 per 100,000) was found in men in the north region. The age-adjusted incidence ratio for men in the north region compared with the south was 1.38 (95 per cent confidence interval 1.34 to 1.42). Similar differences were found in women: incidence ratio for north compared with south region 1.39 (1.07 to 1.81). The proportion treated was larger in men and varied by region: 46.9 per cent of men in the mid region compared with 43.7 per cent in the south received treatment (P < 0.001), whereas 29.8 per cent of women in the north region versus 25.4 per cent in the south had an intervention (P = 0.001). The incidence did not increase over time. CONCLUSION: The higher incidence of AAA in the north of Sweden corresponds well with reported CHD patterns. The incidence of AAA in the population did not increase significantly over time, in contrast to the increasing intervention rates.

The state of the art of osseointegration for limb prosthesis.

Osseointegration (OI) is the direct attachment of bone onto a titanium implant. Recently, the term is used to describe "transdermal" implants that allow an external prosthesis to be connected directly to the skeleton. This technology eliminates the challenges of conventional socket-based prostheses, such as skin breakdown and poor fit, which are common in patients with major extremity amputations. Osseointegration patients demonstrate encouraging improvements in quality of life and function. Patients report improvement in prosthetic use, prosthetic mobility, global health, and pain reduction on a variety of clinical assessment tools. Various implants have been developed for osseointegration for amputees. These implants use a variety of fixation strategies and surface augments to allow for successful integration into the host bone. Regardless of design, all OI implants face similar challenges, particularly infections. Other challenges include the inability to determine when integration has occurred and the inability to detect loss of integration. These challenges may be met by incorporating sensing systems into the implants. The percutaneous nature of the metal devices can be leveraged so that internal sensors need not be wireless, and can be interrogated by external monitoring systems, thus providing crucial, real-time information about the state of the implant. The purpose of this review is to (1) review the basic science behind osseointegration, (2) provide an overview of current implants, practice patterns, and clinical outcomes, and (3) preview sensor technologies which may prove useful in future generations of transdermal orthopaedic implants.

Mirna biogenesis pathway is differentially regulated during adipose derived stromal/stem cell differentiation.

Stromal/stem cell differentiation is controlled by a vast array of regulatory mechanisms. Included within these are methods of mRNA gene regulation that occur at the level of epigenetic, transcriptional, and/or posttranscriptional modifications. Current studies that evaluate the posttranscriptional regulation of mRNA demonstrate microRNAs (miRNAs) as key mediators of stem cell differentiation through the inhibition of mRNA translation. miRNA expression is enhanced during both adipogenic and osteogenic differentiation; however, the mechanism by which miRNA expression is altered during stem cell differentiation is less understood. Here we demonstrate for the first time that adipose-derived stromal/stem cells (ASCs) induced to an adipogenic or osteogenic lineage have differences in strand preference (-3p and -5p) for miRNAs originating from the same primary transcript. Furthermore, evaluation of miRNA expression in ASCs demonstrates alterations in both miRNA strand preference and 5'seed site heterogeneity. Additionally, we show that during stem cell differentiation there are alterations in expression of genes associated with the miRNA biogenesis pathway. Quantitative RT-PCR demonstrated changes in the Argonautes (AGO1-4), Drosha, and Dicer at intervals of ASC adipogenic and osteogenic differentiation compared to untreated ASCs. Specifically, we demonstrated altered expression of the AGOs occurring during both adipogenesis and osteogenesis, with osteogenesis increasing AGO1-4 expression and adipogenesis decreasing AGO1 gene and protein expression. These data demonstrate changes to components of the miRNA biogenesis pathway during stromal/stem cell differentiation. Identifying regulatory mechanisms for miRNA processing during ASC differentiation may lead to novel mechanisms for the manipulation of lineage differentiation of the ASC through the global regulation of miRNA as opposed to singular regulatory mechanisms.

System for in situ studies of atmospheric corrosion of metal films using soft x-ray spectroscopy and quartz crystal microbalance.

We present a versatile chamber ("atmospheric corrosion cell") for soft x-ray absorption/emission spectroscopy of metal surfaces in a corrosive atmosphere allowing novel in situ electronic structure studies. Synchrotron x rays passing through a thin window separating the corrosion cell interior from a beamline vacuum chamber probe a metal film deposited on a quartz crystal microbalance (QCM) or on the inside of the window. We present some initial results on chloride induced corrosion of iron surfaces in humidified synthetic air. By simultaneous recording of QCM signal and soft x-ray emission from the corroding sample, correlation between mass changes and variations in spectral features is facilitated.

Caspase-2: an orphan enzyme out of the shadows.

Caspase-2 has been embodied as an initiator or executioner protease in diverse apoptotic scenarios. However, accumulating evidence is challenging this view, pertaining to its true role. The enzyme's catalytic activity is currently implicated in various functions required for correct cell proliferation, such as counteracting genomic instability, as well as suppressing tumorigenesis. Here, apart from summarizing the latest observations in caspase-2-related research, we make an attempt to reconcile these findings and discuss their implications for future directions.

Estrogen effects on sister chromatid exchanges in mouse uterine cervical and kidney cells.

Uterine cervical fibroblasts and kidney cells from 3-to 4-day-old, previously untreated or phenobarbital [(PBA) CAS: 50-06-6]-treated female NMRI mice were cultured in vitro in the presence of diethylstilbestrol [(DES) CAS: 56-53-1] or 17 beta-estradiol [(E2) CAS: 50-28-2]. DES at 10(-7) M in the culture medium increased the number of sister chromatid exchanges (SCE); no further increases was obtained with higher concentrations of DES. Pretreatment of the females with PBA resulted in an increase in SCE when DES was used in the range of 10(-7) to 10(-5) M. The level of this increase was significantly higher than that in fibroblasts from untreated females. E2 had no effect. DES had no effect on SCE in kidney cells from PBA-treated females. Indomethacin and alpha-naphthoflavone reduced the number of SCE in PBA-exposed fibroblasts to an intermediate level. The presence of both enzyme inhibitors in the medium depressed the number of SCE to the control level.

3-methylcholanthrene: transient inhibition of the lytic step of mouse natural killer cells.

Female mice belonging to NMRI stock were inoculated neonatally with daily doses of 5 micrograms diethylstilbestrol (DES) or olive oil for the first 5 days after birth and in adult life with 20 or 100 micrograms 3-methylcholanthrene (MCA) or the vehicle tricaprylin only. The cytotoxic activity of spleen natural killer (NK) cells against YAC-1 target cells was studied in a 51Cr release assay at 2, 5, 10, or 15 days after the MCA injection. A dose of 20 micrograms MCA to neonatally olive oil-injected females did not influence the NK activity, whereas an injection of 100 micrograms MCA significantly depressed the NK activity to about half the value seen in controls. This suppression was transient, and the normal level was reached again 15 days after the injection. The depressed NK activity could not be related to humoral or cellular suppressor mechanisms. A study at the single-cell level revealed that the inhibition was due to interference with the lytic step of the NK cell without affecting target-binding capacity. In vitro exposure of spleen cells from MCA-treated animals to interferon fully restored the NK activity. Neonatal DES treatment resulted in a depressed NK activity in adult females to a level about half of that seen in olive oil-injected controls. The NK activity in DES-treated females was not influenced by either 20 or 100 micrograms MCA. The MCA-induced suppression of NK activity was discussed in relation to the earlier reported difference in incidence of MCA-induced sarcomas between DES-treated and control females after they were given 20 micrograms MCA in adult life, as well as in relation to the same incidence in the 2 groups treated with 100 micrograms MCA. The results are compatible with a significant role for NK cells during MCA carcinogenesis and indicate that a possible part of the tumorigenic effect of MCA is its early suppressive effect on NK cell activity.

Long-term effects of neonatal estrogen treatment on mitogen responsiveness of mouse spleen lymphocytes.

Neonatal female NMRI mice were given injections of olive oil (controls) or daily doses of corticosterone (10 micrograms), 17 beta-estradiol (10 micrograms), or diethylstilbestrol (DES) (0.01, 0.1, 1, or 5 micrograms) for the first 5 days after birth. The 5-micrograms dose of DES resulted in a persistently reduced in vitro mitogen response to concanavalin A or bacterial lipopolysaccharide of spleen lymphocytes from 6-, 10-, and 18-week-old or 17-month-old females. DES injections from day 6 through day 10 did not influence the later mitogen response. Treatment of ovariectomized 10-week-old females with 5 micrograms DES for 5 days resulted in a tendency to a reduced mitogen response (not statistically significant) 24 hours after the last DES injection. Four weeks later, the mitogen response was the same in experimental and control females. Different possible mechanisms for the persistent effect on the mitogen response are discussed. Neonatal DES treatment not only resulted in persistent changes in the cervicovaginal epithelium and in the hypothalamic-pituitary gland control system but also in the spleen lymphocyte mitogen response. The altered mitogen response should be a stimulus for a detailed analysis of the immune system in women exposed to DES during fetal life, some of whom develop later in life clear cell adenocarcinoma of the uterine cervix and vagina.

Molecular recognition in thylakoid structure and function.

In photosynthesis, light-harvesting chlorophyll molecules are shunted between photosystems by phosphorylation of the protein to which they are bound. An anchor for the phosphorylated chlorophyll-protein complex has now been identified in the reaction centre of chloroplast photosystem I. This finding supports the idea that molecular recognition, not membrane surface charge, governs the architecture of the chloroplast thylakoid membrane. We describe a model for the chloroplast thylakoid membrane that is consistent with recent structural data that specify the relative dimensions of intrinsic protein complexes and their dispositions within the membrane. Control of molecular recognition accommodates membrane stacking, lateral heterogeneity and regulation of light-harvesting function by means of protein phosphorylation during state transitions--adaptations that compensate for selective excitation of photosystem I or photosystem II. High-resolution structural description of membrane protein-protein interactions is now required to understand thylakoid structure and regulation of photosynthesis.

Neonatal estrogen treatment and epithelial abnormalities in the cervicovaginal epithelium of adult mice.

Female NMRI mice were given injections of different doses of 17 beta-estradiol, 17 alpha-estradiol, diethylstilbestrol (DES), dienestrol, trans-stilbene, progesterone, testosterone, 5 alpha-dihydrotestosterone, or olive oil for the first 5 days after birth. When the females were killed at 8 weeks after birth, all the estrogens, effective at different dose levels (10(-2) to 5 microgram/day), had resulted in the display by several of the cervicovaginal preparations studied of a heterotopic columnar epithelium (HCE) in regions where females given injections of olive oil, testosterone, 5 alpha-dihydrotestosterone, progesterone, or trans-stilbene had only the normal squamous epithelium. The further fate of the HCE was followed at two later age stages, 36 to 52 weeks and 14 to 17 months. The HCE developed into glandular-like structures penetrating into the stroma and justifying the designation of adenosis. DES resulted in a more pronounced adenosis than did 17 beta-estradiol; in both cases, metaplasia was a striking component of the adenosis regions. Development of adenosis from HCE was dependent upon presence of the ovaries. Some preparations from 44-week-old females given DES injections showed signs of a beginning malignant transformation in the adenosis regions, more evident in 17-month-old females. Among the 23 preparations in the latter group, 8 had changes morphologically indicating malignancy with examples of adenocarcinoma, mixed carcinoma, and squamous carcinoma. Because of the seemingly low aggressive nature of this malignancy, the term "pseudocarcinoma" is discussed. Ten- to 12-week-old BALB/c and C57BL/6 females given DES injections neonatally had HCE in the uterine cervix and vaginal fornices after neonatal DES injections. Differences in extension of HCE were observed after DES injections for three different 5-day periods in th neonatal and immature stages of NMRI females. An interaction between different DES-sensitive parameters to result in the pseudocarcinomas is discussed.

Natural killer cell activity and tumor susceptibility in female mice treated neonatally with diethylstilbestrol.

Female NMRI or AKR/J mice were given daily s.c. injections of 5 micrograms diethylstilbestrol (DES) in 0.025 ml olive oil, or of olive oil only, for the first 5 days after birth. At the age of 6 to 7 weeks, both DES-treated females and control females were killed, and the cytotoxic activity of the spleen cells against standard natural killer cell target YAC-1 cells as well as the natural killer cell-sensitive I-522 cells and relatively insensitive I-51 AKR lymphomas were tested. The cytotoxic activity against I-51 cells was similar for DES-treated and control females while the DES-treated females had only about one-half the cytotoxicity activity to I-522 and YAC-1 cells as did controls. Control females eliminated radioactivity derived from 125I-labeled YAC-1 and I-522 target cells injected i.v. faster than did DES-treated females, while the results were similar for both animal groups when using I-51 cells. The cumulative death incidence was higher for DES-treated females than for control females after incubation with low numbers of I-522 cells but similar for both groups when using I-51 cells. Finally, the incidence of females developing methylcholanthrene-induced sarcomas, using a simple low-dose injection (10 or 20 micrograms), was higher among DES-injected animals than among controls. Taken together, the results indicate that female mice treated neonatally with DES have a functionally defective natural killer cell population, resulting in increased tumor susceptibility.

Caspase-2: the reinvented enzyme.

On the basis of evidences that caspase-2 gene targeting in several generated mouse models accelerates tumor formation, this enzyme was recently implicated in tumor suppression. The observed function, however, compels other molecular perturbations harboring tumorigenic properties. Therefore, the question remains as to whether or not caspase-2 can be considered a true tumor suppressor? The traditional view of caspase-2 being vital for the apoptotic response to induced cell stress in some systems is in line with these findings. Yet, caspase-2 has also been associated with other processes which equally might interfere with tumorigenic potential, including the oxidative stress response, aging and genome surveillance. By different mechanisms, this enzyme has been proposed to function as a checkpoint regulator in the cell cycle. Together, these data indicate that caspase-2 is a highly versatile factor, a view that is contrasted by the alternative explanation where the enzyme harbors a mechanism affecting a discrete process, which in turn is functionally connected to other cell systems. In any case, it is clear that the general view of caspase-2 as a protein mainly involved in apoptotic cell death is shattered. Hence, we wish to discuss the perspectives of recent achievements in caspase-2-related research.

If your lump is bigger than a golf ball and growing, think Sarcoma.

AIM: Only 1 in 100 of primary care consultations regarding new soft tissue lumps (STL) are malignant and are susceptible to a delay in diagnosis. We aimed to generate a Bayesian Belief Network to estimate the likelihood of malignancy in patients to facilitate the initial evaluation of a STL and improve timing and quality of referrals to specialist treatment centres. METHODS: We evaluated all patients referred with a new STL between 1996 and 2007. Variables investigated focused on patient factors, symptoms and STL characteristics. Relevant data was extracted and coded for statistical analysis. RESULTS: 3018 patients with a STL were assessed, of which 1563 (52%) were benign and 1455 (48%) malignant. The features most conditionally associated with the outcome of interest (Benign or Malignant) are referred to as first-degree associates, and are increasing size, age, size of the lump, and duration of symptoms, in that order. On cross validation, this model demonstrated an AUC of 0.77 (95%C.I. 0.75-0.79). CONCLUSIONS: For the first time, we have described the hierarchal relationship between factors and created an aide memoire, larger than a golf ball and growing, to trigger referral to tertiary tumor units. Importantly, we found pain to be a poor discriminatory factor. We hope our findings will lead to greater awareness and earlier diagnosis of STL.

HELIOS--A laboratory based on high-order harmonic generation of extreme ultraviolet photons for time-resolved spectroscopy.

In this paper, we present the HELIOS (High Energy Laser Induced Overtone Source) laboratory, an in-house high-order harmonic generation facility which generates extreme ultraviolet (XUV) photon pulses in the range of 15-70 eV with monochromatized XUV pulse lengths below 35 fs. HELIOS is a source for time-resolved pump-probe/two-color spectroscopy in the sub-50 fs range, which can be operated at 5 kHz or 10 kHz. An optical parametric amplifier is available for pump-probe experiments with wavelengths ranging from 240 nm to 20,000 nm. The produced XUV radiation is monochromatized by a grating in the so-called off-plane mount. Together with overall design parameters, first monochromatized spectra are shown with an intensity of 2 ⋅ 10(10) photons/s (at 5 kHz) in the 29th harmonic, after the monochromator. The XUV pulse duration is measured to be <25 fs after monochromatization.

The development of a rat model to investigate the formation of blast-related post-traumatic heterotopic ossification.

Currently, there is no animal model in which to evaluate the underlying physiological processes leading to the heterotopic ossification (HO) which forms in most combat-related and blast wounds. We sought to reproduce the ossification that forms under these circumstances in a rat by emulating patterns of injury seen in patients with severe injuries resulting from blasts. We investigated whether exposure to blast overpressure increased the prevalence of HO after transfemoral amputation performed within the zone of injury. We exposed rats to a blast overpressure alone (BOP-CTL), crush injury and femoral fracture followed by amputation through the zone of injury (AMP-CTL) or a combination of these (BOP-AMP). The presence of HO was evaluated using radiographs, micro-CT and histology. HO developed in none of nine BOP-CTL, six of nine AMP-CTL, and in all 20 BOP-AMP rats. Exposure to blast overpressure increased the prevalence of HO. This model may thus be used to elucidate cellular and molecular pathways of HO, the effect of varying intensities of blast overpressure, and to evaluate new means of prophylaxis and treatment of heterotopic ossification.

The content of a specific cell product in the vaginal epithelium of normal and neonatally estrogenized mice: its dependence on an estradiol-prolactin interaction.

The amount of an immunological marker (CVA) in mucified cells of the mouse vaginal epithelium was quantified by a mixed hemagglutination technique for tissue sections. Immature mice, adult mice which had been estrogenized neonatally (5 mug diethylstilbestrol daily for the first five days after birth), and adult non-estrogenized mice were studied. All adult animals were castrated 7-10 days before starting the experiments. Injections of 5 mug estradiol-17beta (48 and 24 h before killing the animals) increased the amount of CVA in all three groups of animals, but most markedly in the neonatally estrogenized mice. The amount of CVA found following estradiol treatment was decreased in adult animals injected with the ergot alkaloid CB154 (0.5 mg twice daily for 6 days) in addition to the hormone. This partial block of the estradiol-induced CVA response by CB154 was relieved by exogenous rat prolactin. The CVA content in immature animals was not influenced by CB154, given alone or together with estradiol. Combined treatment with estradiol and rat prolactin (3 mug every 8 h for 6 days) increased more efficiently than estradiol alone the amount of CVA in immature and adult nonestrogenized animals. Prolactin injected alone had no effect on the CVA content. These data strongly suggest a synergistic action of estradiol and prolactin in augmenting the epithelial CVA content. Explants of the vaginal wall from normal and neonatally estrogenized mice were grafted into the thigh muscles of newborn mice, every host carrying one graft from both types of animals. The CVA content in the epithelium of the two grafts increased to the same level in response to estradiol. When the hosts were injected with estradiol and prolactin, the CVA content was higher in grafts from estrogenized donors than in those from nonestrogenized animals. Our results demonstrate that the mucified vaginal cells in adult, neonatally estrogenized mice have a content of CVA which is higher than in nonestrogenized animals. This difference may be ascribed to hormonal factors (estradiol-prolactin) as well as to persistent effects in the vaginal cells as a result of the neonatal estrogen treatment.

Delayed hypersensitivity response to oxazolone in neonatally estrogenized mice.

Neonatal NMRI mice were treated with diethylstilbestrol (DES) fof the first 5 days after birth. At 6 and 9 months the same animals were tested for delayed hypersensitivity reaction using an ear test with oxazolone. The DES treated animals had a diminished response to oxazolone compared with controls.

Protein tyrosine phosphorylation in the transition to light state 2 of chloroplast thylakoids.

Redox dependent protein phosphorylation in chloroplast thylakoids regulates distribution of excitation energy between the two photosystems of photosynthesis, PS I and PS II. Several thylakoid phosphoproteins are known to be phosphorylated on N-terminal threonine residues exposed to the chloroplast stroma. Phosphorylation of light harvesting complex II (LHC II) on Thr-6 is thought to account for redistribution of light energy from PS II to PS I during the transition to light state 2. Here, we present evidence that a protein tyrosine kinase activity is required for the transition to light state 2. With an immunological approach using antibodies directed specifically towards either phospho-tyrosine or phospho-threonine, we observed that LHC II became phosphorylated on both tyrosine and threonine residues. The specific protein tyrosine kinase inhibitor genistein, at concentrations causing no direct effect on threonine kinase activity, was found to prevent tyrosine phosphorylation of LHC II, the transition to light state 2, and associated threonine phosphorylation of LHC II. Possible reasons for an involvement of tyrosine phosphorylation in light state transitions are proposed and discussed.

New approaches for estimating risk from exposure to diethylstilbestrol.

A subgroup from a National Institute of Environmental Health Sciences, workshop concerned with characterizing the effects of endocrine disruptors on human health at environmental exposure levels considered the question, If diethylstilbestrol (DES) were introduced into the market for human use today and likely to result in low-dose exposure of the human fetus, what would be required to assess risk? On the basis of an analysis of the quality of data on human DES exposure, the critical times and doses for inducing genital tract malformations and cancer must be determined. This would be facilitated through analysis of the ontogeny of estrogen receptor expression in the developing human genital tract. Models of low-dose estrogenic effects will have to be developed for human and rodent genital tract development. Mouse models offer many advantages over other potential animal models because of the wealth of the earlier literature, the availability of sensitive end points, the availability of mutant lines, and the possibility of generating genetically engineered model systems. Through multidisciplinary approaches, it should be possible to elucidate the cellular and molecular mechanisms of endocrine disruption elicited by estrogens during development and facilitate an assessment of risk to humans.