ShapeCut: Bayesian surface estimation using shape-driven graph.
A variety of medical image segmentation problems present significant technical challenges, including heterogeneous pixel intensities, noisy/ill-defined boundaries and irregular shapes with high variability. The strategy of estimating optimal segmentations within a statistical framework that combines image data with priors on anatomical structures promises to address some of these technical challenges. However, methods that rely on local optimization techniques and/or local shape penalties (e.g., smoothness) have been proven to be inadequate for many difficult segmentation problems. These challenging segmentation problems can benefit from the inclusion of global shape priors within a maximum-a-posteriori estimation framework, which biases solutions toward an object class of interest. In this paper, we propose a maximum-a-posteriori formulation that relies on a generative image model by incorporating both local and global shape priors. The proposed method relies on graph cuts as well as a new shape parameters estimation that provides a global updates-based optimization strategy. We demonstrate our approach on synthetic datasets as well as on the left atrial wall segmentation from late-gadolinium enhancement MRI, which has been shown to be effective for identifying myocardial fibrosis in the diagnosis of atrial fibrillation. Experimental results prove the effectiveness of the proposed approach in terms of the average surface distance between extracted surfaces and the corresponding ground-truth, as well as the clinical efficacy of the method in the identification of fibrosis and scars in the atrial wall.


Impaired function of endothelial progenitor cells in children with primary systemic vasculitis.
Previously, we demonstrated that children with active systemic vasculitis (SV) have higher circulating CD34 + CD133 + KDR+ endothelial progenitor cells (EPC); the function of these EPCs, and their relationship with disease activity in vasculitis remains largely unexplored. We hypothesized that although EPC numbers are higher, EPC function is impaired in active SV of the young. The aims of this study were therefore to: 1. investigate the relationship between disease activity and EPC function in children with SV; and 2. study the influence of systemic inflammation on EPC function by investigating the effects of hyperthermia and TNF-α on EPC function. We performed a cross-sectional study of unselected children with SV with different levels of disease activity attending a single center (Great Ormond Street Hospital, London) between October 2008 and December 2014. EPCs were isolated from peripheral blood of children with SV, and healthy child controls. EPC function was assessed by their potential to form colonies (EPC-CFU), and ability to form clusters and incorporate into human umbilical vein endothelial cell (HUVEC) vascular structures in matrigel. The effects of hyperthermia and TNF-α on EPC function were also studied. Twenty children, median age 12-years (5-16.5; nine males) were studied. EPC-CFU and the number of EPC clusters formed on matrigel were significantly reduced in children with active vasculitis compared with healthy controls (p = 0.02 for EPC-CFU; p = 0.01 for EPC cluster formation). Those with active vasculitis had lower EPC-CFU and EPC cluster formation than those with inactive disease, although non-significantly so. In addition, EPC incorporation into matrigel HUVEC networks was lower in children with SV compared with healthy children, irrespective of disease activity. Ex-vivo pre-treatment of EPC with hyperthermia impaired EPC function; TNF-α down-regulated EPC expression of CD18/CD11b and resulted in decreased incorporation into HUVEC networks. Whilst our previous work showed that circulating CD34 + EPC numbers are well preserved, this study revealed that EPC function is significantly impaired in children with vasculitis. It is possible that the chronic inflammatory milieu associated with vasculitis may impair EPC function, and thus contribute to an unfavourable balance between endothelial injury and repair. The mechanism of this remains to be established, however.


Tumor necrosis factor-α-induced apoptosis of gastric cancer MKN28 cells: accelerated degradation of the inhibitor of apoptosis family members.
The role of the inhibitor of apoptosis (IAP) family members in tumor necrosis factor-α (TNF-α)-induced apoptosis of human gastric cancer MKN28 cells was explored. TNF-α induced up-regulation of cIAP2, whereas cycloheximide (CHX) induced down-regulation of XIAP and survivin. Degradation of cIAP1 and XIAP, but not survivin, was accelerated by co-treatment of cells with TNF-α and CHX, and TNF-α-induced up-regulation of cIAP2 was inhibited by BMS-345541 (NF-κB inhibitor). Treatment of MKN28 cells with TNF-α plus CHX induced degradation of survivin and activation of caspase-8 and -3, followed by degradation of cIAP1 and XIAP and apoptosis. Proteasome inhibitors (MG132 and epoxomicin) suppressed TNF-α plus CHX-induced degradation of survivin, cIAP1, and XIAP as well as apoptosis. A caspase inhibitor (z-VAD-fmk) suppressed TNF-α plus CHX-induced apoptosis, but allowed degradation of survivin, cIAP1 and XIAP. TNF-α receptor 1 and 2 were expressed on MKN28 cells. The magnitude of apoptosis induced by TNF-α plus BMS-345541 was much less than that induced by TNF-α plus CHX. These findings suggest that TNF-α plus CHX-induced apoptosis of gastric cancer MKN28 cells may be caused by accelerated degradation of the IAP family members (survivin, cIAP1, and XIAP), in addition to inhibition of NF-κB-dependent synthesis of anti-apoptotic molecules.


[Pyomyositis of the M. quadriceps femoris in an 8-month-old boy following pneumococcal meningitis].
An 8-month-old boy developed pyomyositis of the M. quadriceps femoris following meningitis caused by Streptococcus pneumoniae. The child had no underlying immune defect. Conservative treatment with antibiotics led to complete clinical and radiological resolution of the infection. In Europe, pyomyositis is rarely seen in children. Pyomyositis should be considered in the differential diagnosis in children with fever and complaints of joint pain or myalgia. Ultrasound and MRI are helpful techniques for establishing the diagnosis and for differentiating pyomyositis from other pathological conditions. Treatment consists of adequate antibiotics and in some cases drainage.


Post-thaw application of ROCK-inhibitors increases cryopreserved T-cell yield.
Emerging cell-based therapies such as CAR-T (Chimeric Antigen Receptor T) cells require cryopreservation to store and deliver intact and viable cells. Conventional cryopreservation formulations use DMSO to mitigate cold-induced damage, but do not address all the biochemical damage mechanisms induced by cold stress, such as programmed cell death (apoptosis). Rho-associated protein kinases (ROCK) are a key component of apoptosis, and their activation contributes to apoptotic blebbing. Here we demonstrate that the ROCK inhibitor fasudil hydrochloride, when supplemented into the thawing medium of T-cells increases the overall yield of healthy cells. Cell yield was highest using 5 or 10% DMSO cryopreservation solutions, with lower DMSO concentrations (2.5%) leading to significant physical damage to the cells. After optimisation, the post-thaw yield of T-cells increased by approximately 20% using this inhibitor, a significant increase in the context of a therapy. Flow cytometry analysis did not show a significant reduction in the relative percentage of cell populations undergoing apoptosis, but there was a small reduction in the 8 hours following thawing. Fasudil also led to a reduction in reactive oxygen species. Addition of fasudil into the cryopreservation solution, followed by dilution (rather than washing) upon thaw also gave a 20% increase in cell yield, demonstrating how this could be deployed in a cell-therapy context, without needing to change clinical thawing routines. Overall, this shows that modulation of post-thaw biochemical pathways which lead to apoptosis (or other degradative pathways) can be effectively targeted as a strategy to increase T-cell yield and function post-thaw.


Histologic examination of selected areas of canine pulmonary arteries.
Selected areas of pulmonary arteries from 18 healthy mixed-breed dogs were examined using histologic staining techniques. Smooth muscle cell, collagen, and elastin content of the tunica intima and tunica media were assessed. Fifteen dogs had abnormalities of tunica intima or tunica media in at least one arterial section examined. Of all arterial sections examined, 40% had histologic changes of the tunica intima or tunica media, and 42% of these vascular lesions were in the main pulmonary artery. The most commonly occurring pathologic change was loss of smooth muscle cells and elastin of the tunica media and replacement by collagen. This lesion is similar to cystic medionecrosis of the aorta. Seemingly, a high frequency of spontaneous vascular lesions exist in pulmonary arteries of young dogs.


An administration of TAK-683 at a minimally effective dose for luteinizing hormone stimulation under the absence of the ovary induces luteinizing hormone surge in ovary-intact goats.
The present study aimed to evaluate hormonal responses and their association with the TAK-683 blood concentrations in goats administered TAK-683 at a low dose, which had been previously determined as the minimally effective dose for luteinizing hormone (LH) stimulation in ovariectomized goats. In Experiment 1, 5 µg of TAK-683 treatment had no significant stimulatory effect on LH secretion in ovariectomized Shiba goats (n = 4). In Experiment 2, cycling goats received the treatment of prostaglandin F2α and progesterone-releasing controlled internal drug releasing (CIDR) to induce the follicular phase, then they were treated with 5 µg of TAK-683 (hour 0) intravenously (n = 4, IV) or subcutaneously (n = 3, SC) or with vehicle intravenously (n = 4, control) at 12 h after CIDR removal. Blood samples were collected at 10-min (-2-6 h), 2-h (6-24 h), or 6-h (24-48 h) intervals. Ovarian ultrasonographic images were assessed daily to confirm ovulation after the treatment. A surge-like release of LH was immediately observed after injection in all animals in the IV (peak time: 4.2 ± 0.6 h, peak concentration: 73.3 ± 27.5 ng/ml) and SC (peak time: 4.6 ± 0.4 h, peak concentration: 62.6 ± 23.2 ng/ml) groups, but not in the control group. Ovulation was detected within 3 days after TAK-683 injection in all animals in the IV and SC groups, and the interval period from TAK-683 administration to ovulation in the IV group was significantly (P < 0.05) shorter than that of the control group. No significant changes were observed between the IV and SC groups in terms of luteal diameter and blood progesterone levels after ovulation. The present findings suggest that the involvement of one or more ovarian factor(s) is indispensable for a TAK-683-induced LH surge leading to ovulation in goats.


Arterial and venous abdominal thrombosis in a 79-year-old woman with COVID-19 pneumonia.
As coronavirus pandemic continue to spread over the world, we have to be aware of potential complications on hospitalized patients. We report a case of a 79-year-old woman with COVID-19 pneumonia complicated by combined arterial and venous thrombosis of upper mesenteric vessels. As unenhanced chest CT scan plays a key role in managing the COVID-19 pandemic, we should pay attention to indirect signs of thrombosis.


Size matters! A reliable endovascular method to avoid infolding and endoleaks when reconstructing aortic bifurcations using stent grafts.
Conventional revascularization of the lower extremities secondary to aortoiliac atherosclerotic vascular disease (ASVD) involves major intra-abdominal surgery with aortobifemoral bypass grafting. Complication rates are as high as 10% and perioperative mortality can be as high as 5%; therefore, careful patient selection is paramount. Endovascular aortobifemoral bypass repair with aortic bifurcation reconstruction is an emerging option in patients deemed unsuitable for conventional surgical management. Emerging techniques such as snorkel/chimney method are expanding the number of patients who can undergo endovascular repair of abdominal aortic aneurysms and/or revascularization of the lower extremities. However, until now, objective criteria for stent size selection used in these hybrid techniques have not been described.


Comparison of different commercial DNA extraction kits and PCR protocols for the detection of Echinococcus multilocularis eggs in faecal samples from foxes.
Effective and sensitive methods for the molecular detection of Echinococcus multilocularis in faecal samples of final hosts are crucial for the prevention and control of human alveolar echinococcosis and for studies on the epidemiology of the parasite. Little is known about the suitability of commercial test kits for isolating DNA of E. multilocularis from fox faeces and the performance of standard Polymerase Chain Reaction (PCR) protocols in relation to the quality of DNA extracted by these kits. We compared four different kits: ZR Faecal DNA MiniPrep™ (Zymo Research), FastDNA® SPIN Kit for Soil (MP Biomedicals), QIAamp® Fast DNA Stool Mini Kit (QIAGEN) and NucleoSpin® Soil Kit (Macherey-Nagel) for the extraction of DNA from E. multilocularis eggs present in faeces of foxes. Negative faecal samples were spiked with 600, 300, 150, 75, 37, 18, 9, 5 or 2 E. multilocularis eggs, and each egg concentration was tested 10 times with each of the DNA extraction kits. Each extracted DNA sample was amplified using three PCR protocols: i. conventional PCR (cPCR, Platinum®Taq, Invitrogen), ii. qPCR with the iQ™ Supermix (Bio-Rad) and iii. qPCR with the QuantiTect® Multiplex-Master Mix (QIAGEN). The highest analytical sensitivities for molecular detection of E. multilocularis eggs in spiked fox faeces were observed when combining either the QIAamp® Fast DNA Stool Mini Kit or the ZR Faecal DNA MiniPrep™ kit with the qPCR using the QuantiTect® Multiplex-Master Mix (Sensitivities 97% and 94%, respectively). Combinations including the remaining test kits (NucleoSpin® Soil Kit and FastDNA® SPIN Kit for Soil) showed a markedly lower analytical sensitivity for PCR examinations. The results of the present study indicate that it is of utmost importance to select suitable DNA extraction kits in combination with robust PCR methods or reagents to achieve acceptable analytical sensitivity in the molecular detection of E. multilocularis eggs in fox faecal samples.


[Elastic stockings and compression therapy, arguments favoring more rational use].
Elastic stockings and compression therapy are often considered as a major tool in the prevention and in the treatment of chronic venous insufficiency and of lymphedema. Nevertheless we must note that this therapeutic option is both underused and misused. We discuss the reasons for this paradox and the expected modes of action and pathophysiological benefits, and finally propose a prescription guide (indications, dosage, practice details) for a rational use of elastic stockings and compression therapy.


Cloning and expression of the Bacteroides fragilis YCH46 neuraminidase gene in Escherichia coli and Bacteroides uniformis.
A neuraminidase-encoding gene nanH of Bacteroides fragilis strain YCH46 was cloned into the cosmid vector pHC79. The nanH gene was subcloned from the cosmid and was located within a 2.2-kb XhoI-KpnI fragment. Southern hybridization experiments demonstrated that the gene was present as a single copy on the bacterial chromosome. Neuraminidase activity expressed in the initial Escherichia coli clone was approximately 3600-fold lower than that expressed in B. fragilis YCH46. However, when nanH was transferred from E. coli to B. uniformis by mobilization of a shuttle plasmid, the transconjugant expressed 1100-fold higher activity than the E. coli donor did. These results suggest that modes of nanH expression in E. coli and Bacteroides are heterologous.


Does the existence of HCMV components predict poor prognosis in glioma?
Human cytomegalovirus (HCMV) has been associated with malignant gliomas. The objective of the present study was to further investigate the existence and prognostic value of HCMV components in gliomas. Using immunohistochemical staining, HCMV proteins IE1-72 and pp65 were examined in 67 glioma specimens of various histologic grades, in comparison to 6 -control brain tissue samples. The HCMV DNA was measured in both the tumor tissues and the peripheral blood of the patients, using nested PCR. Kaplan-Meier analysis and Cox proportional hazards models were used to analyzed the prognostic value of HCMV components in glioma tissues. IE1-72 was detected in 76.1% (51/67) of glioma tissues, and pp65 was detected in 65.7% (44/67) of glioma tissues. HCMV DNA was detected in 52.2% (35/67) of glioma tissues and 29.9% (20/67) of peripheral blood samples of glioma patients. These HCMV components were not detected in control brain tissue. However, the existence of HCMV components showed no significant correlation with the prognosis of glioma patients. Our results demonstrate that although HCMV proteins and nucleic acids are present in gliomas, they do not correlate with the prognosis. The role of HCMV in gliomas needs to be carefully interpreted.


Athletes Perceived Level of Risk Associated with Botanical Food Supplement Use and Their Sources of Information.
Athletes should carefully consider the use of botanical food supplements (BFSs) given the current lack of substantiation for botanical nutrition and health claims under EU and UK food laws. In addition, athletes may be at an increased risk of doping violations and other adverse outcomes potentially associated with BFS use; however, little is known about athletes' intake, knowledge, or perceptions in relation to BFS use. An online cross-sectional survey of n = 217 elite and amateur athletes living on the island of Ireland was conducted using Qualtrics XM to assess intake, knowledge, attitudes, and perceptions. General food supplements (FSs) were reported by approximately 60% of the study cohort, and 16% of the supplements reported were categorized as BFS. The most frequently consumed BFSs were turmeric/curcumin (14%), Ashwagandha (10%), and Beetroot extract (8%). A higher proportion of amateur athletes would source information about BFSs from less credible sources, such as fellow athletes, or from internet sources or their coach, compared to elite athletes. Those who sourced information about botanicals from fellow athletes (p = 0.03) or the internet (p = 0.02) reported a lower perceived level of risks associated with BFS use. This study therefore suggests that amateur athletes may be more likely to source information from less credible sources compared to elite athletes who may have more access to nutrition professionals and their knowledge/advice. This may have potential adverse implications for amateur athletes, e.g., Gaelic games players, who are included within the doping testing pool but who may not have access to evidence-based nutrition advice.


PMesogenin1 and 2 function directly downstream of Xtbx6 in Xenopus somitogenesis and myogenesis.
T-box transcription factor tbx6 and basic-helix-loop-helix transcription factor pMesogenin1 are reported to be involved in paraxial mesodermal differentiation. To clarify the relationship between these genes in Xenopus laevis, we isolated pMesogenin2, which showed high homology with pMesogenin1. Both pMesogenin1 and 2 appeared to be transcriptional activators and were induced by a hormone-inducible version of Xtbx6 without secondary protein synthesis in animal cap assays. The pMesogenin2 promoter contained three potential T-box binding sites with which Xtbx6 protein was shown to interact, and a reporter gene construct containing these sites was activated by Xtbx6. Xtbx6 knockdown reduced pMesogenin1 and 2 expressions, but not vice versa. Xtbx6 and pMesogenin1 and 2 knockdowns caused similar phenotypic abnormalities including somite malformation and ventral body wall muscle hypoplasia, suggesting that Xtbx6 is a direct regulator of pMesogenin1 and 2, which are both involved in somitogenesis and myogenesis including that of body wall muscle in Xenopus laevis.


[Study on bovine spongiform encephacitis and current status of safety control of medical devices deriving from animals].
To introduce the basic information about mad cow disease and the current status of safety control of medical devices derived from mammalian animal tissues. Literature concerned was reviewed intensively. Mad cow disease also known as bovine spongiform encephacitis (BSE), is generally considered from the same source of Scrapie, and they are caused by the same kind of sponginess brain tissue pathological changes. Mad cow disease is caused by the misfolding of a small protein called Prion. This disease has the character of slowly breaking down the central neuron system of animals. Further researches can provide evaluation for safety considerations of medical devices deriving from animal.


Free Will, Determinism, and Intuitive Judgments About the Heritability of Behavior.
The fact that genes and environment contribute differentially to variation in human behaviors, traits and attitudes is central to the field of behavior genetics. Perceptions about these differential contributions may affect ideas about human agency. We surveyed two independent samples (N = 301 and N = 740) to assess beliefs about free will, determinism, political orientation, and the relative contribution of genes and environment to 21 human traits. We find that lay estimates of genetic influence on these traits cluster into four distinct groups, which differentially predict beliefs about human agency, political orientation, and religiosity. Despite apparent ideological associations with these beliefs, the correspondence between mean lay estimates and published heritability estimates for the surveyed traits is large (r = .77). Belief in genetic determinism emerges as a modest predictor of accuracy in these lay estimates. Additionally, educated mothers with multiple children emerge as particularly accurate in their estimates of the genetic contribution to these traits.


A retrospective comparison of BMI changes and the potential risk factors among schizophrenic inpatients treated with aripiprazole, olanzapine, quetiapine or risperidone.
The objective of this study was to evaluate weight gain and its potential risk factors among different second generation antipsychotics (SGAs). The study was conducted for Korean inpatients with schizophrenia in a university hospital in Seoul, between Jan 2000 and Dec 2007. Data were collected by reviewing the medical records of the patients, who were prescribed to one of the SGAs among aripiprazole, olanzapine, quetiapine or risperidone. The changes of weight and body mass index (BMI); risk of clinically significant weight gain (>7% gain to initial weight) and their associations with various clinical characteristics of such patients were analyzed. Five hundred and eighty-eight (588) and 294 subjects treated with one of the four SGAs for a duration of 1 month and 2 months were included, respectively. Olanzapine showed significantly greater weight and BMI increase at month 1 (p=0.028 for weight; p=0.019 for BMI) and month 2 (p=0.032 for weight; p=0.029 for BMI) than others. Females showed greater BMI increase change (0.70±0.91 kg/m(2), p=0.008) and were also more likely to experience clinically significant weight gain (odd ratio=1.846, 95% CI=1.098 to 3.105, p=0.021) at month 1. Younger patients (<45 years old) had significantly greater weight and BMI increase at both months 1 and 2. Younger patients also showed greater risk for clinically significant weight gain at month 2 (odd odd ratio=2.567, 95% CI=1.196 to 5.508, p=0.016). Low baseline BMI (<25 kg/m(2)) was associated with greater weight gain at month 1 (1.92±2.29 kg, p<0.001) and month 2 (4.07±3.56 kg, p<0.001) and BMI increase at month 1 and month 2 (p<0.001 for both). Patients with low baseline BMI showed higher risk of clinically significant weight gain at both months 1 and 2 (p<0.001 for both). Olanzapine was shown to have higher metabolic risk than other SGAs in inpatients with schizophrenia. The individual's own clinical characteristics also exerted influence on weight gain effects of SGAs. Younger patients with lower baseline BMI were under greater risk of antipsychotic-induced weight gain. More studies are required to verify the role of gender on weight gain.


[Effect of actinomycin D and cycloheximide on lipoproteins in blood serum and liver cytosol of rats under oxidative stress].
The influence of protein synthesis inhibitors on rat lipoprotein content and composition under oxidative stress caused by cobalt chloride injection has been investigated in the present work. It has been concluded that apoprotein synthesis is very important process influencing on adaptive reactions under free-radical oxidation activation conditions. Co-administration of cobalt chloride and actinomycin D or cycloheximide (the inhibitors of the protein synthesis) has realy prevented hyperlipoproteinemia in many cases but hasn't influenced on lipoprotein oxidation. Pre-beta- and beta-lipoproteins were discussed to have mRNA pool in hepatocytes.


An Initiative to Change Inpatient Practice: Leveraging the Patient Medical Home for Postdischarge Follow-Up.
The standard of care for hospital discharge planning includes arranging follow-up appointments, usually with a primary care provider. However, follow-up phone calls instead of face-to-face visits may be an appropriate alternative for some patients. This option was explored within the framework of the US Department of Veterans Affairs (VA) patient-centered medical home model of care, the Patient Aligned Care Team. At a VA hospital, a pilot study was conducted on the use of phone calls from members of a patient's medical home as posthospital discharge follow-up rather than the traditional face-to-face provider model. Inpatient providers were educated about the phone follow-up alternative, and this option was standardized as part of discharge planning rounds. During Phase 1 at one clinic over three months, 17 of 118 eligible patients received phone call follow-up (14.4% of discharges) instead of traditional face-to-face follow-up. During Phase 2, data from Phase 1 were analyzed, and staff at the other eight clinic sites were trained. After the expansion of the initiative to all regional clinic sites in Phase 3, 76 of 447 eligible discharges (17.0%) were scheduled for phone follow-up. As a balancing metric, there were no significant differences in rates of 30-day emergency department (ED) utilization (11.9% and 5.9%, (p = 0.47)) or nonelective rehospitalization (16.8% and 17.6%, (p = 0.93)) between these groups during Phase 1. This initiative changed provider practices to use phone call follow-up for select patients instead of face-to-face provider visits after hospital discharge, without significantly increasing rates of 30-day ED utilization or rehospitalization.


COP1 regulates the stability of CAM7 to promote photomorphogenic growth.
The unique member of the calmodulin gene family, Calmodulin7 (CAM7), plays a crucial role as transcriptional regulator to promote Arabidopsis seedling development. CAM7 regulates the expression of HY5, which is intimately involved in the promotion of photomorphogenic growth and light-regulated gene expression. COP1 ubiquitin ligase suppresses photomorphogenesis by degrading multiple photomorphogenesis promoting factors including HY5 in darkness. Genetic interaction studies, in this report, reveal that CAM7 and COP1 co-ordinately work to promote photomorphogenic growth and light-regulated gene expression at lower intensity of light. CAM7 physically interacts with COP1 in the nucleus. Further, in vivo study suggests that CAM7 and COP1 interaction is light intensity dependent. We have also shown that functional COP1 is required for optimum accumulation of CAM7 at lower fluences of light. Taken together, this study demonstrates the coordinated function of CAM7 and COP1 in Arabidopsis seedling development.


Our experience with surgical management of patients with diffuse coronary artery disease.
During a follow up period of 5 years (January 1983-December 1988), 145 consecutive patients (14% of all patients undergoing coronary artery bypass surgery) underwent multiple coronary artery bypass grafting combined with endarterectomy whenever necessary for treatment of severe diffuse triple-vessel coronary artery disease. Fifty-one patients (35%) had poor left ventricular ejection fraction (less than 35%). Associated left main coronary artery disease was present in 30 (21%) patients. All coronary arteries and branches with greater than 50% obstructive disease were bypassed, using saphenous vein conduit; average grafts per patient were 5.5. Endarterectomies were done in 137 patients in 210 vessels. Right coronary artery was the commonest site (132 patients). Multiple vessel endarterectomy (greater than 2 vessels) was done in 44 patients (30%). The peri-operative mortality was 3.5%. Pre-operatively, 75% patients had class III and 14% class IV (Canadian Cardiovascular Society angina criteria). During the mean follow up period of 2 years, 86% patients have class 1 symptoms, and 14% have class II symptom. Thirty of these 145 patients have undergone resting and exercise radionuclide ventriculographic studies which have shown a significant improvement in the ejection fraction response to exercise (p less than 0.05). Thus, patients with severe diffuse coronary disease can undergo multiple bypass grafting procedure, along with endarterectomies with low mortality rates and improved exercise tolerance and functional classification.


Influence of Temperature on the Efficacy of Homologous and Heterologous DNA Vaccines against Viral Hemorrhagic Septicemia in Pacific Herring.
Homologous and heterologous (genogroup Ia) DNA vaccines against viral hemorrhagic septicemia virus (genogroup IVa) conferred partial protection in Pacific Herring Clupea pallasii. Early protection at 2 weeks postvaccination (PV) was low and occurred only at an elevated temperature (12.6°C, 189 degree days), where the relative percent survival following viral exposure was similar for the two vaccines (IVa and Ia) and higher than that of negative controls at the same temperature. Late protection at 10 weeks PV was induced by both vaccines but was higher with the homologous vaccine at both 9.0°C and 12.6°C. Virus neutralization titers were detected among 55% of all vaccinated fish at 10 weeks PV. The results suggest that the immune response profile triggered by DNA vaccination of herring was similar to that reported for Rainbow Trout Oncorhynchus mykiss by Lorenzen and LaPatra in 2005, who found interferon responses in the early days PV and the transition to adaptive response later. However, the protective effect was far less prominent in herring, possibly reflecting different physiologies or adaptations of the two fish species. Received August 1, 2016; accepted March 10, 2017.


Comparison of enzyme kinetic parameters obtained in vitro for reactions mediated by human CYP2C enzymes including major CYP2C9 variants.
Cytochrome P450 (CYP) 2C enzymes contribute to the metabolism of about 30% of all drugs. Known polymorphisms of the respective enzymes and drug-drug interactions have a major impact on the efficacy and safety of some CYP2C substrate drugs. In vivo - in vitro correlations including prediction of the effect of such covariates requires quantitative information on enzyme kinetics. In this article there will be a summary of the values of the Michaelis-Menten constant (K(m)), the maximal velocity (V(max)) and the intrinsic clearance (Cl(int); V(max)/K(m)) for 84 substrates (100 reactions) reported to be mediated by CYP2C9 (variant enzymes CYP2C9.1, CYP2C9.2 and CYP2C9.3), CYP2C8 and/or CYP2C19. Particularly contradictory findings for the same reactions call for some standardization in the assessment of enzyme kinetics.


Outpatient grip strength measurement predicts survival, perioperative adverse events, and nonhome discharge among patients with vascular disease.
Frailty is associated with adverse outcomes among patients with vascular disease. Grip strength measurement is a comparatively simple, quick, and inexpensive screening test for weakness (a component of frailty) that is potentially applicable to clinical practice. We hypothesized that grip strength and categorical weakness are associated with clinical outcomes among patients with vascular disease. To test this hypothesis, we conducted a longitudinal cohort study evaluating associations between grip strength measured during outpatient clinic visits for vascular disease and clinical outcomes, including survival and perioperative outcomes. Adult patients recruited from outpatient vascular surgery and/or vascular medicine clinics underwent dominant hand grip strength measurement using a hand dynamometer. Participants were categorized as weak based on grip strength, sex, and body mass index. Multivariable logistic models were used to evaluate perioperative outcomes. Mortality was evaluated using Cox proportional hazards models adjusted for sex, age, and operative intervention during follow-up. We enrolled 321 participants. The mean patients age was 69.0 ± 9.4 years, and 33% were women. Mean grip strength was 32.0 ± 12.1 kg, and 92 participants (29%) were categorized as weak. The median follow-up was 24.0 months. Adverse perioperative events occurred in 32 of 84 patients undergoing procedures. Grip strength was associated with decreased risk of perioperative adverse events (hazard ratio [HR], 0.41 per 12.7 kg increase; 95% confidence interval [CI], 0.20-0.85; P = .0171) in a model adjusted for open versus endovascular procedure (HR, 12.75 for open; 95% CI, 2.54-63.90; P = .0020) and sex (HR, 3.05 for male; 95% CI, 0.75-12.4; P = .120). Grip strength was also associated with a lower risk of nonhome discharge (HR, 0.34 per 12.7 kg increase; 95% CI, 0.14-0.82; P = .016) adjusted for sex (HR, 2.14 for male; 95% CI, 0.48-9.50; P = .31) and open versus endovascular procedure (HR, 10.36 for open; 95% CI, 1.20-89.47; P = .034). No associations between grip strength and length of stay were observed. Mortality occurred in 48 participants (14.9%) during follow-up. Grip strength was inversely associated with mortality (HR, 0.46 per 12.5 kg increase; 95% CI, 0.29-0.73; P = .0009) in a model adjusted for sex (HR, 5.08 for male; 95% CI, 2.1-12.3; P = .0003), age (HR, 1.04 per year; 95% CI, 1.01-1.08), and operative intervention during follow-up (HR, 1.23; 95% CI, 0.71-2.52). Categorical weakness was also associated with mortality (HR, 1.81 vs nonfrail; P = .048) in a model adjusted for age (HR, 1.06 per year; P = .002) and surgical intervention (HR, 1.36; 95% CI, 1.02-0.09; P = .331). Grip strength is associated with all-cause mortality, perioperative adverse events, and nonhome discharge among patients with vascular disease. These observations support the usefulness of grip strength as a simple and inexpensive risk screening tool for patients with vascular disease.


Snapping Pes Syndrome after Unicompartmental Knee Arthroplasty.
Snapping pes syndrome is defined as a snapping sensation in the medial knee caused by pes anserinus and rarely occurs. Snapping pes syndrome after unicompartmental knee arthroplasty (UKA) has not been reported yet. We experienced two cases with this syndrome after UKA. Conservative treatment was effective in one case, while surgical excision of the gracilis tendon was necessary to relieve painful snapping in the other case. The main cause of the first case might be posteromedial overhang of the tibial tray that reached up to 5 mm. The probable cause of the second case was posteromedial overhang of the mobile bearing.


Performance of Endophyte Infected Tall Fescue in Europe and North America.
Human assisted plant invasions from Europe to North America have been more common than the reverse. We tested endophyte-mediated performance of tall fescue in parallel three year experiments in Europe and the USA using endophyte infected and uninfected wild and cultivated plants. Experimental plants were subjected to nutrient and water treatments. Whereas endophyte infection increased tall fescue performance in general, the effects of endophytes on plant growth and reproduction varied among plant origins under different environmental conditions. Naturally endophyte-free Finnish cultivar 'Retu' performed equally well as 'Kentucky-31' in both geographic locations. All Eurasian origin plants performed well in the US. In Finland, plants established well and both cultivars survived over the first winter. However, winter mortality of 'Kentucky-31' plants was higher, particularly in fertilized soils in the subsequent winters. Our results suggest that tall fescue ecotype 'Kentucky-31' that flourishes in North America is poorly adapted to Northern European conditions.


Formative evaluation of home visitors' role in addressing poor mental health, domestic violence, and substance abuse among low-income pregnant and parenting women.
This research assessed home visitor effectiveness in communicating about and responding to poor mental health, domestic violence, and substance abuse among pregnant and parenting women home visited as part of a comprehensive family support strategy in seven urban communities. Cross-sectional studies were conducted with mothers (n = 189) actively engaged in home visitation programs and home visitors (n = 45). Maternal interviews assessed need for and receipt of mental health, domestic violence, and substance abuse services, and home visitor discussion of these risk areas. Home visitor surveys assessed perceived adequacy of training and personal effectiveness in addressing these risk areas. Over half of mothers needed mental health, domestic violence, or substance abuse services; however, only 27% of mothers in need of service received services. Most mothers reported having communicated with their home visitor about the three risk areas, but there were no differences in communication frequency based on whether services were needed. Most home visitors perceived themselves as effective in communicating about and responding to these risk factors but rated the training they had received in these areas as less than adequate. Home visitors could benefit from more intensive training in the formal assessment of risks and the protocols for communication about those risks with their clients. Home visitors could also receive support from and work in collaboration with professionals in addressing client risks. Further research on home visit content is needed to determine which strategies facilitate home visitors' ability to effectively communicate about and address client risks.


Increased Cellular Presence After Sciatic Neurectomy Improves the Bone Mechano-adaptive Response in Aged Mice.
The mechano-adaptive response of bone to loading in the murine uniaxial tibial loading model is impaired in aged animals. Previous studies have shown that in aged mice, the amount of bone formed in response to loading is augmented when loads are applied following sciatic neurectomy. The synergistic effect of neurectomy and loading remains to be elucidated. We hypothesize that sciatic neurectomy increases cellular presence, thereby augmenting the response to load in aged mice. We examined bone adaptation in four groups of female C57BL/6J mice, 20-22 months old: (1) sham surgery + 9N loading; (2) sciatic neurectomy, sacrificed after 5 days; (3) sciatic neurectomy, sacrificed after 19 days; (4) sciatic neurectomy + 9N loading. We examined changes in bone cross sectional properties with micro-CT images, and static and dynamic histomorphometry with histological sections taken at the midpoint between tibiofibular junctions. The response to loading at 9N was not detectable with quantitative micro-CT data, but surface-specific histomorphometry captured an increase in bone formation in specific regions. 5 days following sciatic neurectomy, the amount of bone in the neurectomized leg was the same as the contralateral leg, but 19 days following sciatic neurectomy, there was significant bone loss in the neurectomized leg, and both osteoclasts and osteoblasts were recruited to the endosteal surfaces. When sciatic neurectomy and loading at 9N were combined, 3 out of 4 bone quadrants had increased bone formation, on the endosteal and periosteal surfaces (increased osteoid surface and mineralizing surface respectively). These data demonstrate that sciatic neurectomy increases cellular presence on the endosteal surface. With long-term sciatic-neurectomy, both osteoclasts and osteoblasts were recruited to the endosteal surface, which resulted in increased bone formation when combined with a sufficient mechanical stimulus. Controlled and localized recruitment of both osteoblasts and osteoclasts combined with appropriate mechanical loading could inform therapies for mechanically-directed bone formation.


N,N-dimethylformamide-induced modulation of organ- and tumor-associated markers in cultured human colon carcinoma cells.
Cultured human colon carcinoma cells were induced by the polar solvent N,N-dimethylformamide (DMF) to express a more differentiated phenotype as indicated by three types of antigen markers. Carcinoembryonic antigen expression in all DMF-induced cell lines was enhanced. Exposure of cells to DMF resulted in a reduction in ability to bind antibody to tumor-derived colon mucoprotein antigen (CMA) with a concomitant gain in binding of antibody to normal CMA. DMF-induced differentiation was further indicated by modulation of blood group antigen expression. DMF treatment decreased the amount of expression of the H-gene determinant. When DMF-treated cells were cultured in the absence of DMF for 14 days, the levels of expression of the antigen markers reverted to those characteristic of their untreated counterparts.


Effect of plane of nutrition on adipose tissue lipid metabolism in genetically obese and lean pigs.
Genetically lean and obese pigs of 27 kg body weight were individually fed at varying levels of intake. During the first 5 weeks (period 1) the pigs on the high plane nutrition (H) gained 19 kg, those on the medium plane (M) gained 7 kg and those on the low plane (L) lost 5 kg. During the second 5 weeks (period 2) the M pigs gained an additional 7 kg (MM), whereas the period 1 H pigs were fed to lose 5 kg (HL) and the period 1 L pigs were fed to gain 19 kg (LH). All pigs were targeted to weigh 41 kg at the end of 10 weeks. Subcutaneous adipose tissue samples were obtained by biopsy from each pig on day 28 and day 63 of the experiment, i.e., after 4 weeks of feeding a particular level of intake in each period. In vitro glucose metabolism to CO2, total lipids and glyceride fatty acids as well as basal and epinephrine-stimulated lipolytic rates were assessed. Obese pigs had greater cell size and lipogenic and lipolytic rates than lean pigs. During both periods the glucose metabolism rates paralleled the plane of nutrition, i.e., the low intake level yielded low rates and the high intake level yielded high rates. The lipolytic activities were refractory to the plane of nutrition. Finally the less muscular, obese and more muscular, lean pigs presented similar qualitative metabolic responses to their planes of nutrition, although the quantitative responses were divergent.


Microphase separation in multigraft copolymer melts studied by random-phase approximation and self-consistent field theory.
Microphase separation of AB multigraft copolymers with various numbers of graft arms per junction along the backbone was examined by the random-phase approximation and real-space implemented self-consistent field theory. The calculations carried out show that the number of graft arms per junction exerts a marked effect on the phase behavior of the multigraft copolymers. The spinodal shows an upward shift with increasing number of graft arms per junction. The order-order transitions shift toward the higher volume fraction of backbone and the ordered region becomes narrower when the number of graft arms per junction increases. The influence of the number of graft arms per junction on the domain size and the interfacial width has also been examined. It was found that the characteristic domain size decreases and the interfacial width broadens with increasing number of graft arms per junction. The theoretical results at a strong segregation were compared with the existing experimental observations and a good agreement is shown.


Are you the problem?
Many CEOs have a blind spot when pushing for culture change in their organizations; they fail to grasp that they may well be the biggest obstacle to change. Self-reflection is not among the traits that drive executives to the top spot, but once there, they need to be aware of how their management style, for better or worse, sets the tone for the organization as a whole.


Effect of agricultural land-use change on ant dominance hierarchy and food preferences in a temperate oak forest.
The discovery-dominance trade-off is the inverse relationship between the ability of a species to discover resources and the species' dominance of those resources; a paradigm used to explain species coexistence in ant communities dependent on similar resources. However, factors such as stress (e.g., temperature) or disturbance (e.g., removal of biomass) associated with the change in land use, can modify this trade-off. Here, we aimed to determine the potential effects of land use change on dominance hierarchy, food preferences and on the discovery-dominance trade-off. An experiment with baits was used to investigate the dominance hierarchies of ant communities in a temperate mountain habitat in central Mexico. We evaluated the dominance index (DI), food preferences and discovery-dominance trade-offs of ants inhabiting two types of vegetation: a native oak forest and agricultural land resulting from agricultural land use and grazing. The ant communities in both environments were comprised of three species of ants (Monomorium minimum, Myrmica mexicana, and Camponotus picipes pilosulus), four morphospecies (Pheidole sp.1 and Pheidole sp.2, Temnothorax sp. and Lasius sp.) and one genus (Formica spp.). All Formicidae showed values of intermediate to low DI, and this factor did not seem to be influenced by the change in land use. Ants in the modified vegetation (i.e., agricultural land) were found to be numerically greater. Overall, a higher number of visits were registered to the tuna bait, although the duration of foraging events to the honey baits was longer. However, foraging times were dependent on the species considered: the generalized Myrmicinae, M. minimum, the ant species with highest DI, foraged for longer periods of time in the agricultural land and on the tuna bait. Meanwhile, the cold-climate specialist Formica spp., with a lower DI, foraged for longer periods of time in the oak (although not significant) and on the honey bait. We found little evidence of the discovery-dominance trade-off; instead, we found considerable diversity in the strategies used by the different species to access resources. This range of strategies is well represented by the generalized Myrmicinae M. minimum, the cold-climate specialists Formica spp. and Temnothorax sp., and the rare species, as the cold climate specialist Lasius sp. (insinuators). Our evaluation shows that transformation of the original habitat does not appear to affect the hierarchical dominance of the ant communities, but it does affect their food preferences. Species with higher DI values such as the generalized Myrmicinae are more skilled at resource acquisition in modified habitats. Our results suggest that change in land use promotes an increase in the diversity of foraging strategies used by different ant species. This diversity may contribute to resource partitioning which favors coexistence.


Nanostructure-Directed Chemical Sensing: The IHSAB Principle and the Effect of Nitrogen and Sulfur Functionalization on Metal Oxide Decorated Interface Response.
The response matrix, as metal oxide nanostructure decorated n-type semiconductor interfaces are modified in situ through direct amination and through treatment with organic sulfides and thiols, is demonstrated. Nanostructured TiO₂, SnOx, NiO and CuxO (x = 1,2), in order of decreasing Lewis acidity, are deposited to a porous silicon interface to direct a dominant electron transduction process for reversible chemical sensing in the absence of significant chemical bond formation. The metal oxide sensing sites can be modified to decrease their Lewis acidity in a process appearing to substitute nitrogen or sulfur, providing a weak interaction to form the oxynitrides and oxysulfides. Treatment with triethylamine and diethyl sulfide decreases the Lewis acidity of the metal oxide sites. Treatment with acidic ethane thiol modifies the sensor response in an opposite sense, suggesting that there are thiol (SH) groups present on the surface that provide a Brønsted acidity to the surface. The in situ modification of the metal oxides deposited to the interface changes the reversible interaction with the analytes, NH₃ and NO. The observed change for either the more basic oxynitrides or oxysulfides or the apparent Brønsted acid sites produced from the interaction of the thiols do not represent a simple increase in surface basicity or acidity, but appear to involve a change in molecular electronic structure, which is well explained using the recently developed inverse hard and soft acids and bases (IHSAB) model.


Thermochromic discotic 6-oxoverdazyls.
6-Oxoverdazyls 1b[n], substituted with three 3,4,5-trialkoxyphenyl groups (n = 8 and 10), exhibit a columnar hexagonal phase (Col(h)) below 130 °C. They display reversible color change from red to green at the transition to the isotropic phase. XRD and magnetization data do not support dimerization of the radicals in the mesophase.


High-yield synthesis of amido-functionalized polyoctahedral oligomeric silsesquioxanes by using acyl chlorides.
Homosubstituted amido-functionalized polyoctahedral oligomeric silsesquioxanes (POSS) have been synthesized by using acyl chlorides in high yields (ca. 95%). The method proved to be superior over "conventional" syntheses applying carboxylic acids or acid anhydrides, which are much less efficient (ca. 60% yield). A palette of aryl and alkyl groups has been used as side-chains. The structures of the resulting amide-POSS are supported by multinuclear (1)H, (13)C, (29)Si NMR and FTIR spectroscopy and their full conversion into octasubstituted derivatives was confirmed using mass spectrometry. We also demonstrate that the functionalized silsesquioxanes with bulky organic side-chains attached to cubic siloxane core form spherical-like, well-separated nanoparticles with a size of approximately 5 nm.


Integration of the information from gene expression and metabolic fluxes for the analysis of the regulatory mechanisms in Synechocystis.
Synechocystis was grown under autotrophic, mixotrophic and heterotrophic conditions, and the gene expression patterns at the mRNA and protein levels were measured by using semi-quantitative RT-PCR and two-dimensional electrophoresis (2DE), respectively. Moreover, the intracellular metabolic flux distributions in Synechocystis grown under different trophic conditions were also determined using the carbon isotope labeling technique. By combining the information obtained from the transcript levels, protein abundance and metabolic fluxes, the regulatory mechanisms of some enzymes involved in the central metabolism of Synechocystis during growth in the different culture conditions were analyzed. It was found that depending on the energy source available to cyanobacterial cells, the enzymes required for central metabolism were differently regulated according to different mechanisms. The expression of several genes, such as rbcLS and gap2, was light-regulated transcriptionally, while the gene gnd was regulated in response to an apparent flux requirement but by an unknown mechanism. The expression of other genes was independent of the presence of light. The reactions catalyzed by G6PDH, Fbp, PfkA and FbaA were not regulated through enzyme synthesis but by a change in metabolite concentrations. The enzyme PrK was post-translationally regulated by light, probably through the operation of ferredoxin/thioredoxin system. For the enzyme RubisCO, both transcriptional and post-translational regulation was observed. These findings demonstrate that the information obtained from the analysis of mRNA expression, protein expression, and metabolic flux distribution is necessary to understand the regulatory events in complex cellular networks.


Polypeptide nature of growth requirement in yeast extract for Thermoplasma acidophilum.
The active component(s) in yeast extract required by Thermoplasma acidophilum for growth is polypeptide in nature. A fraction from yeast extract was isolated and partially characterized as one or more peptides of molecular weight about 1,000 containing 8 to 10 amino acids. Although it was composed largely of basic and dicarboxylic amino acids, only one amino group per molecule was free. The polypeptide(s) appeared to bind avidly to cations. No other organic compounds except glucose were needed by Thermoplasma. Among several hundred known compounds tested, only glutathione plus Fe2+ or Fe3+, clostridial ferredoxin, and spinach ferredoxin elicited any growth response.


Parasites of carnivorous mammals in Białowieza Primeval Forest.
Although the parasitofauna of wild carnivorous mammals in Poland is quite well recognized, there has been only one research carried on this issue so far in Białowieza Forest--the last lowland primeval forest of temperate zone in Europe. Twelve wild and two domestic species of carnivorous mammals are living in Białowieza Forest. In our work faeces or intestines of all of them except ermine (Mustela erminea) have been examined and parasites or their eggs (or oocysts) recorded. In total 222 faecal probes from 13 species and 11 intestines of weasels have been investigated. All species of examined carnivorous mammals were infected with parasites. The most infected species has occurred red fox Vulpes vulpes (over 70% infected with parasites) meanwhile only about 30% of otters Lutra lutra were infected. We found at least three protozoan species, one trematode, one tapeworm and nine species of nematodes. Except trematode Alaria alata, all of them are reported for the first time from Białowieza Primeval Forest. Oocysts of coccidia have been found in faeces of nine host species, with the highest prevalence (29.4%) in badger. Six species of carnivorous mammals have been occurred infected with trematodes (highest prevalence 42.1% in wolves) and two with tapeworms (Diphyllobothrium latum with the prevalence 2.6% in otter and 31.6% in wolves). All examined host species were infected with nematodes (with prevalence from 14.7% in lynx to 72.7% in red fox). Results of our work should be treated as preliminary researches on the species composition of parasitic fauna invading carnivorous mammals in Białowieza Primeval Forest.


RNA processing is a limiting step for murine tumor necrosis factor beta expression in response to interleukin-2.
We have previously reported that tumor necrosis factor beta (TNF beta) expression is induced by interleukin-2 (IL-2) in the murine lymphocytic T-cell line CTLL-2. In this study, we have characterized the nuclear and cytoplasmic TNF beta transcript and assessed their role in TNF beta gene expression. A unique feature of TNF beta expression was the accumulation of nuclear precursors, which reflected a slow nuclear RNA processing. As a consequence, there was a delay in the appearance of cytoplasmic messengers after the transcriptional induction of TNF beta by IL-2. We also found that two messengers, the fully spliced messenger and an intron 3-retaining messenger, were exported to the cytoplasm and actively translated. The same pattern of expression was observed in concanavalin A-stimulated splenocytes, although the level of expression was much lower than in CTLL-2 cells. The simple genetic structure and the high level of accumulation of nuclear precursors make TNF beta a particularly attractive model system to use for studies of RNA processing and cytoplasmic transport of partially spliced messengers.


Inflammatory Myofibroblastic Tumor: Findings on 68Ga-DOTA-NOC Positron-emission Tomography-Computed Tomography.
Inflammatory myofibroblastic tumor (IMT) or inflammatory pseudotumors are a rare group of tumors usually affecting children and young adults. They occur in various anatomic locations, with most common being the lung accounting for almost 95% of the cases. We present a case of a 4-year-old girl who presented with fever and dull abdominal pain for 6 months with a clinically palpable left hypochondrium mass. On suspicion of mesenteric/gastric neuroendocrine tumor, 68Ga-DOTA-NOC positron-emission tomography-computed tomography was done; however, the biopsy revealed IMT.


Doxorubicin-Conjugated Heparin-Coated Superparamagnetic Iron Oxide Nanoparticles for Combined Anticancer Drug Delivery and Magnetic Resonance Imaging.
In this study, superparamagnetic iron oxide (SPIO) nanoparticles (NPs) with an average size of 10±2 nm were coated with doxorubicin (Dox)-conjugated heparin (DH-SPIO) and were used for targeted anticancer drug delivery, and as a magnetic resonance imaging (MRI) contrast agent. The DH-SPIO NPs had a mean particle size of 125±10 nm and a zeta potential of –35±3 mV. Fourier transform-infrared spectroscopy, X-ray diffraction spectroscopy, transmission electron microscopy, vibrating sample magnetometry, and MTT assay were used to investigate the properties of DH-SPIO NPs. The internalization of DH-SPIO NPs into A549 tumor cells was examined using fluorescence microscopy and quantified by flow cytometry. Prussian blue staining, total iron assay, in vitro MRI and transmission electron microscopy showed that DH-SPIO NPs had high superparamagnetic clustering effect. In vivo therapy of A549 human lung carcinoma, DHSPIO NPs displayed a higher efficacy than Dox in inhibiting tumor growth and prolonging the survival of mice bearing tumors. Meanwhile, the pathological damage to the cardiac tissue in mice treated with DH-SPIO NPs was significantly less severe than that of mice treated with free Dox at the same dosage. These results show that DH-SPIO NPs are promising biomaterials for combined drug therapy and clinical imaging.


Treatment of stroke in aged male and female rats with Vepoloxamer and tPA reduces neurovascular damage.
Stroke is a leading cause of death and disability worldwide, mainly affecting the elderly. Unfortunately, current treatments for acute ischemic stroke warrant improvement. To date, tissue plasminogen activator (tPA) is of limited use in stroke patients mainly due to its narrow therapeutic window and potential for hemorrhagic complication. The adjuvant treatment with Vepoloxamer, a purified amphipathic polymer has been shown to enhance the thrombolytic efficacy of tPA treatment in young adult male rats after embolic stroke. However, most stroke patients are aged; therefore, the current study investigated the therapeutic effect of the combined tPA and Vepoloxamer treatment in aged male and female rats subjected to embolic stroke. Male and female Wistar rats at 18 months of age were subjected to embolic middle cerebral artery occlusion and treated either with monotherapy of tPA or Vepoloxamer, a combination of these two agents, or saline at 4 h after stroke onset. Neurological outcomes were evaluated with a battery of behavioral tests including adhesive removal, foot-fault, and modified neurological severity score tests at 1 and 7 days after stroke onset, followed by histopathological analysis of infarct volume. Residual clot size and vascular patency and integrity were analyzed. The combination treatment with Vepoloxamer and tPA significantly reduced infarct volume and neurological deficits in male and female rats compared to rats treated with saline and the monotherapies of tPA and Vepoloxamer. While Vepoloxamer monotherapy moderately reduced neurological deficits, monotherapies with tPA and Vepoloxamer failed to reduce infarct volume compared to saline treatment. Furthermore, the combination treatment with tPA and Vepoloxamer accelerated thrombolysis, reduced ischemia and tPA-potentiated microvascular disruption, and concomitantly improved cerebrovascular integrity and perfusion in the male ischemic rats. Combination treatment with tPA and Vepoloxamer at 4 h after stroke onset effectively reduces ischemic neurovascular damage by accelerating thrombolysis and reducing ischemia and tPA potentiated side effects in the aged rats. This funding suggests that the combination treatment with tPA and Vepoloxamer represents a promising strategy to potentially apply to the general population of stroke patients.


Enzyme-Cleaved Bone Marrow Transplantation Improves the Engraftment of Bone Marrow Mesenchymal Stem Cells.
Mesenchymal stem cell (MSC) therapy is a promising approach to curing bone diseases and disorders. In treating genetic bone disorders, MSC therapy is local or systemic transplantation of isolated and in vitro proliferated MSC rather than bone marrow transplantation. Recent evidence showed that bone marrow MSC engraftment to bone regeneration has been controversial in animal and human studies. Here, our modified bone marrow transplantation (BMT) method solved this problem. Like routine BMT, our modified method involves three steps: (i) isolation of bone marrow cells from the donor, (ii) whole-body lethal irradiation to the recipient, and (iii) injection of isolated bone marrow cells into irradiated recipient mice via the tail vein. The significant modification is imported at the bone marrow isolation step. While the bone marrow cells are flushed out from the bone marrow with the medium in routine BMT, we applied the enzymes' (collagenase type 4 and dispase) integrated medium to wash out the bone marrow cells. Then, cells were incubated in enzyme integrated solution at 37°C for 10 minutes. This modification designated BMT as collagenase-integrated BMT (c-BMT). Notably, successful engraftment of bone marrow MSC to the new bone formation, such as osteoblasts and chondrocytes, occurs in c-BMT mice, whereas routine BMT mice do not recruit bone marrow MSC. Indeed, flow cytometry data showed that c-BMT includes a higher proportion of LepR+, CD51+, or RUNX2+ non-hematopoietic cells than BMT. These findings suggested that c-BMT is a time-efficient and more reliable technique that ensures the disaggregation and collection of bone marrow stem cells and engraftment of bone marrow MSC to the recipient. Hence, we proposed that c-BMT might be a promising approach to curing genetic bone disorders. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.


Screening for diabetic retinopathy: developing a standardized photographic protocol for St. John's Hospital.
The management of people with diabetes involves input from many healthcare professionals including doctors, nurses, podiatrists and dieticians. In February 1999 at the request of the Consultant Diabetologist a diabetic screening service was established at St. John's Hospital using digital retinal photography. A preliminary photographic protocol was put into place immediately. In order to establish if the protocol was suitable, or at the level of a national standard, the author carried out an extensive literature search. There proved to be a paucity of published information specifically related to guidelines for diabetic retinopathy screening, using a photographic method. The author therefore issued questionnaires to randomly selected medical photography and diabetic screening units throughout the United Kingdom. The research and results of this study has enabled the provision of an effective and efficient retinopathy screening service. Medical photographers are now recognized as part of the diabetic clinic 'team' that care for patients at St. John's Hospital.


Recent developments in the science of proarrhythmic cardiac safety of new drugs.
Following marketing withdrawals of several drugs due to proarrhythmic safety concerns, the ICH Guidelines S7B and E14 were released in 2005 and have guided pre-approval cardiac safety assessments in multiple regulatory jurisdictions. While this S7B-E14 paradigm has successfully prevented drugs with unanticipated potential for inducing Torsades de Pointes entering the market, it has unintentionally resulted in the termination of development programs for potentially important compounds that could have exhibited a favourable benefit-risk balance. The Comprehensive In vitro Proarrhythmia Assay paradigm is currently attracting considerable attention as a solution to this problem. While much evaluative work in this new paradigm will be conducted in the non-clinical domain, human electrocardiographic assessments will remain an important component of the overall investigational strategy, possibly being conducted in Phase I trials employing exposure-response modelling. This article reviews recent developments in proarrhythmic cardiac safety assessments of new drugs, their rationales, and current limitations.


YH25448, an Irreversible EGFR-TKI with Potent Intracranial Activity in EGFR Mutant Non-Small Cell Lung Cancer.
Given that osimertinib is the only approved third-generation EGFR-TKI against EGFR activating and resistant T790M mutated non-small cell lung cancer (NSCLC), additional mutant-selective inhibitors with a higher efficacy, especially for brain metastases, with favorable toxicity profile are still needed. In this study, we investigated the antitumor efficacy of YH25448, an oral, mutant-selective, irreversible third-generation EGFR-TKI in preclinical models. Antitumor activity of YH25448 was investigated in vitro using mutant EGFR-expressing Ba/F3 cells and various lung cancer cell lines. In vivo antitumor efficacy, ability to penetrate the blood-brain barrier (BBB), and skin toxicity of YH25448 were examined and compared with those of osimertinib using cell lines and PDX model. Compared with osimertinib, YH25448 showed a higher selectivity and potency in kinase assay and mutant EGFR-expressing Ba/F3 cells. In various cell line models harboring EGFR activating and T790M mutation, YH25448 effectively inhibited EGFR downstream signaling pathways, leading to cellular apoptosis. When compared in vivo at equimolar concentrations, YH25448 produced significantly better tumor regression than osimertinib. Importantly, YH25448 induced profound tumor regression in brain metastasis model with excellent brain/plasma and tumor/brain area under the concentration-time curve value. YH25448 rarely suppressed the levels of p-EGFR in hair follicles, leading to less keratosis than osimertinib in animal model. The potent systemic and intracranial activity of YH25448 has been shown in an ongoing phase I/II clinical trial for advanced EGFR T790M mutated NSCLC (NCT03046992). Our findings suggest that YH25448 is a promising third-generation EGFR inhibitor, which may be more effective and better tolerated than the currently approved osimertinib.


Calcium-antagonist effects of quercetin on aortic smooth muscle.
Relaxation induced by quercetin was studied on aortic rings depolarized with KCl and on Ca2+ dependent aortic contraction. Quercetin's actions as a cyclooxygenase inhibitor were compared with those of indomethacin. To determine the possible effects of quercetin on internal Ca2+ mobilization, it was assayed on aortic muscle contracted with caffeine. Quercetin relaxes aortic smooth muscle contracted with KCl (RC50 = 1.8 x 10(-4) M) and induces a rightward displacement of Ca2+ curves. The relaxation induced by quercetin is higher than that produced by indomethacin, suggesting that its action is independent of cyclooxygenase inhibition. Caffeine-induced aortic contraction was not modified by quercetin, suggesting that its effect should not involve internal calcium modulation. Our results further support the thesis that the spasmolytic action of quercetin arises from its negative modulation of calcium entry to the smooth muscle.


[The role of aspiration cytology in tumor diagnostics].
Fine needle aspiration biopsy (FNAB) of focal lesions is a quick, relatively simple and cost-effective diagnostic method. However, performing aspirations and interpreting smears require skill and experience. Before initiating an aspiration the doctor needs to be aware of the limits of cytology as it is vital to know what kind of diagnostic issues can be answered upon a smear and what kind of questions cannot. Traditionally FNAB was performed without radiologic guidance, and therefore almost only palpable lesions were aspirated. Since ultrasound (US) has become widely used in medicine, it is axiomatical that FNAB is ideally performed with US guidance not only for the protection of the patients but also for targeting the lesion more safely. Several cytologists find US guidance unnecessary as a routinely used examination, which may lead to unsatisfactory smears and false negative results. This means not only a loss for the patient, but leads to a negative judgement of this diagnostic method. Our interventional cytology diagnostic team developed a working method resulting in excellent statistical results. In the followings we would like to share our experience refined the past two decades to restore the reputation of this diagnostic method.


Safety of colorado microdissection needle (stryker) for skin opening in craniomaxillofacial surgery.
Diathermy is used widely in surgical procedures, mainly for subcutaneous and deeper layers dissection. The use on craniomaxillofacial skin has been precluded by the fear of scar formation, alopecia and wound dehiscence. The aim of this study was to clinically evaluate the safety of Colorado Microdissection needle (Stryker) for skin opening in craniomaxillofacial surgery. 117 skin incisions for craniomaxillofacial procedures were performed using the Stryker Colorado microdissection needle (CMN). The incisions included Coronal, Hemicoronal, Preauricular, Subciliary, Lateral brow's, Submandibular, Retromandibular, Risdon's, Modified Blair's and Schoebinger's incision. The reason for the craniomaxillofacial operative intervention included: Craniofacial trauma, removal of benign and malignant pathology, craniofacial reconstruction and Temporomandibular joint surgeries. 115 incisions were performed in patients in the adult age group and 02 in the pediatric age group. All the patients tolerated the procedures well, with no increased risk from the use of the CMN. Only one incision had wound infection and dehiscence. All other patients had usual wound healing. At 6 months follow-up postoperative scar and alopecia was evaluated. Only on close up inspection minimal area of alopecia was noticed along the incision line. The postoperative scar was healthy in all the cases. The findings of this study recommend the use of the CMN in all craniomaxillofacial procedures.


[The effect of pemolin on the mitotic activity of Vicia faba L (author's transl)].
The effect of diverse concentrations of 5-phenyl-2-imino-4-oxazolidone (PIO, pemolin, Tradon) on the mitotic activity in lateral roots of Vicia faba L. was studied by aerated and non-aerated hydrocultivation with and without mineral nutrition, respectively. With optimal conditions (aerated nutrient solution) weak PIO-concentrations, most significantly 10(-6) g/ml, effected a marked increase of the mitotic index. Contrarily, strong PIO-concentrations (10(-4) and 3 X 10(-4) g/ml = saturated solution) significantly decreased the mitotic index though simultaneously preserving the mitotic activity in long-term experiments, when on account of nutrient deficiency it had already collapsed in weak PIO-concentrations and the controls. The activating effect of weak PIO-concentrations compared with the controls is more significant in stress situations (nutrient deficiency, O2-deficiency) than under optimal conditions. Furthermore a slight acceleration of mid-mitotic phases (metaphase--anaphase) recognized by a marked decrease in percentage of these phases, can be stated with weak PIO-concentrations, again particularly so with 10(-6) g/ml. In total, dependent on concentration, pemolin presumably may either activate or suppress cell metabolism and particularly the mitotic cycle. The exact site of action of the substance is still unknown.


Application of photoassisted electrochemical detection to explosive-containing environmental samples.
High-performance liquid chromatography (HPLC) with ultraviolet (UV) absorbance and photoassisted electrochemical detection (PAED) is applied to the determination of explosives in groundwater and soil samples. On-line, solid-phase extraction minimizes sample pretreatment, enabling direct analysis of groundwater samples and soil extracts. Soils are extracted using pressurized fluid extraction, which is compared to the Environmental Protection Agency (EPA) sonication method. Limits of detection for explosives in the matrixes of interest are equivalent or superior (i.e., <10 parts-per-trillion for HMX) to those achieved using the EPA method 8330. HPLC-UV-PAED is also shown here to be more broadly applicable, as it is capable of determining nitro compounds of interest (e.g., nitroglycerin) that have poor UV chromophores. Additional selectivity of amine-substituted nitroaromatic explosives is achieved by using a photochemical reactor with a 366-nm wavelength lamp. By coupling reversed-phase columns of different selectivities together, baseline resolution of all 14 standard explosives is demonstrated.


Mitochondrial Dysfunction and Insulin Resistance in Pubertal Youth Living with Perinatally Acquired HIV.
Mitochondrial dysfunction (MD) is linked to cardiometabolic complications, such as obesity and insulin resistance (IR), the frequencies of which are higher in adults living with HIV infection and receiving combination antiretroviral therapies (ARV). ARV-treated youth living with perinatally acquired HIV infection (YLPHIV) may be especially susceptible to IR due to long-term exposure to both factors. Medical histories, fasting blood chemistry panels, and mitochondrial function in banked peripheral blood mononuclear cells (PBMCs) were assessed in eligible YLPHIV from the Pediatric HIV/AIDS Cohort Study (PHACS)/Adolescent Master Protocol (AMP) Mitochondrial Determinants Component cohort, stratified by Homeostatic Model Assessment of IR (HOMA-IR) score: case (score ≥4, n = 39) or control (score <4, n = 105). PBMCs were sources for mitochondrial (mt) DNA copies/cell; mtRNA transcript levels of oxidative phosphorylation (OXPHOS) subunits NADH dehydrogenases 1 and 6, and cytochrome B; and enzymatic activities of OXPHOS Complexes I (CI) and IV (CIV). Logistic regression models were fit to estimate the odds of IR case diagnosis, adjusted for sex, race/ethnicity, body mass index (BMI) z-score, and Tanner stage. IR cases were similar to controls by age, sex, and race/ethnicity. Cases had higher median levels of peak HIV viral load, lactate, pyruvate, triglycerides, and BMI z-scores. OXPHOS CI enzymatic activity was lower in cases (log10 1.62 vs. 1.70) and inversely correlated with HOMA-IR score (r = -0.157, p = .061), but did not associate with IR in adjusted models. Fully adjusted models indicated associations of nadir CD4% [odds ratio (OR) = 0.95, 95% confidence intervals (CIs) = 0.90-1.00] or peak HIV load (OR = 3.48, 95% CIs = 1.70-10.79) with IR. IR in YLPHIV was strongly associated with morphometrics, but early virologic and immunologic factors may also influence MD.


[Chemotherapy of the hormone-refractory prostate cancer].
The scepticism dominating the chemotherapy of the hormone-refractory prostate cancer (HRPC) has been replaced by a wave of enthusiasm. Phase II studies with taxane-containing combination therapies could achieve high response rate in some cases, and HRPC can not longer be deemed resistant to chemotherapy. It remains to be seen whether the combinations offer a survival advantage. This will be tested in phase III studies. Palliative chemotherapy should be considered in patients with HRPC if the initial hormone therapy was effective for a short time only and after several hormone therapies have been completed. Since chemotherapy is not yet an established standard therapy of HRPC, patients should be, if possible, included in clinical studies.


Continuous flow analysis of total organic carbon in polar ice cores.
Ice cores are a widely used archive to reconstruct past changes of the climate system. This is done by measuring the concentration of substances in the ice and in the air of bubbles enclosed in ice. Some species pertaining to the carbon cycle (e.g., CO2, CH4) are routinely measured. However, information about the organic fraction of the impurities in polar ice is still very limited. Therefore, we developed a new method to determine the content of total organic carbon (TOC) in ice cores using a continuous flow analysis (CFA) system. The method is based on photochemical oxidation of TOC and the electrolytic quantification of the CO2 produced during oxidation. The TOC instrument features a limit of detection of 2 ppbC and a response time of 60 s at a sample flow rate of 0.7 mL/min and a linear measurement range of 2-4000 ppbC. First measurements on the ice core from Talos Dome, Antarctica, reveal TOC concentrations varying between 80 and 360 ppbC in the 20 m section presented.


Contact Printing of Multilayered Thin Films with Shape Memory Polymers.
This study describes a method for transfer printing microarrays of multilayered organic-inorganic thin films using shape memory printing stamps and microstructured donor substrates. By applying the films on the microstructured donor substrates during physical vapor deposition and modulating the interfacial adhesion using a shape memory elastomer during printing, this method achieves (1) high lateral and feature-edge resolution and (2) high transfer efficiency from the donor to the receiver substrate. For demonstration, polyurethane-acrylate stamps and silicon/silicon oxide donor substrates were used in the large-area transfer printing of organic-inorganic thin-film stacks with micrometer lateral dimensions and sub-200 nm thickness.


Estimation of heritability and genetic correlation of body weight gain and growth curve parameters in Korean native chicken.
This study estimated the genetic parameters for body weight gain and growth curve parameter traits in Korean native chicken (KNC). A total of 585 F1 chickens were used along with 88 of their F0 birds. Body weights were measured every 2 weeks from hatching to 20 weeks of age to measure weight gain at 2-week intervals. For each individual, a logistic growth curve model was fitted to the longitudinal growth dataset to obtain three growth curve parameters (α, asymptotic final body weight; β, inflection point; and γ, constant scale that was proportional to the overall growth rate). Genetic parameters were estimated based on the linear-mixed model using a restricted maximum likelihood method. Heritability estimates of body weight gain traits were low to high (0.057 to 0.458). Heritability estimates for α, β, and γ were 0.211±0.08, 0.249±0.09, and 0.095±0.06, respectively. Both genetic and phenotypic correlations between weight gain traits ranged from -0.527 to 0.993. Genetic and phenotypic correlation between the growth curve parameters and weight gain traits ranged from -0.968 to 0.987. Based on the results of this study population, we suggest that the KNC could be used for selective breeding between 6 and 8 weeks of age to enhance the overall genetic improvement of growth traits. After validation of these results in independent studies, these findings will be useful for further optimization of breeding programs for KNC.


Rates and relations of mitochondrial genome evolution across the Echinoidea, with special focus on the superfamily Odontophora.
In order to better characterize the placement of genus Tripneustes, as a representative of the Toxopneustidae family within the broader sea urchin mitochondrial (MT) phylogeny, the complete MT genome of Tripneustes gratilla was generated and compared with all published echinoid MT genomes currently available on NCBI GenBank. The MT genome phylogeny supports the existence of the superfamily Odontophora (consisting of the families Strongylocentrotidae, Echinometridae, and Toxopneustidae). A relaxed molecular-clock time calibration suggests a split between the three key Odontophore MT lineages occurred during the late Eocene/Oligocene. Major global oceanographic changes have been inferred during this time frame, potentially driving species diversification through environmental selection pressures. To test for signatures of selection acting on the mitochondria, the historical rate of gene evolution of individual MT genes was assessed through a branch-site comparison of nonsynonymous to synonymous substitution ratios (ω). Models of positive selection and neutral evolution, as compared via a likelihood ratio test, show no evidence of strong historical positive selection on mitochondrial genes at the genesis of the Odontophora. However, while pairwise ω comparison revealed signatures of strong negative selection, relatively elevated ω values were observed within the Strongylocentrotus genus.


Idiopathic congenital talipes equinovarus; not always an isolated anomaly. A review of long-term outcomes.
To describe the long-term outcomes of children at school age who were thought to have isolated idiopathic congenital talipes equino varus (ICTEV) at birth. A retrospective review of all children attending a regional talipes clinic who had achieved school age. One hundred sixty-three children were followed up. ICTEV was more common in Maori and other Polynesian children and in males. Additional abnormalities were apparent in 30.1% overall, but higher in New Zealand Europeans (43%) than Maori (21%) or other Polynesians (22%). Of the abnormalities, 41% were associated with the central nervous system or with neurodevelopmental delay. Conventional G-band karyotyping, where performed, was not informative. The rate of additional abnormalities was higher than previous reports. This has implications for prenatal counseling and postnatal follow-up as a prenatal diagnosis of ICTEV may subsequently be found to have longer term implications in addition to orthopedic treatments of the clubfeet.


Plutonium and radiocerium contents in shallow sea sediment collected off shore in Ibaraki Prefecture, Japan.
The contents of 239, 240Pu and 144Ce in shallow water sediment were measured by means of chemical separation and alpha spectrometry on 239, 240Pu and beta counting on 144Pr . The samples were collected in 1977 and 1980 at off shore of Ibaraki Prefecture for comparison of pre- and after operation of Tokai spent fuel reprocessing plant. The ranges and mean values of 239, 240Pu and 144Ce contents determined in the samples are 0.02 approximately equal to 2.85 Bq/kg-dry (0.55 approximately equal to 77 pCi/kg-dry) with mean values of 0.93 Bq/kg-dry (25 pCi/kg-dry) in 1977 and 0.52 Bq/kg-dry (14 pCi/kg-dry) in 1980 for 239, 240Pu , and 0.67 approximately equal to 19.5 Bq/kg-dry) (18 approximately equal to 526 pCi/kg-dry) with mean value of 5.44 Bq/kg-dry (147 pCi/kg-dry) in 1977 and 1.59 Bq/kg-dry (43 pCi/kg-dry) in 1980 for 144Ce, respectively. These values showed no environmental accumulation of the nuclides by the operation was detected. The contents of two nuclides showed strong correlation each other in the samples. The inventory of 239, 240Pu in the sediment was estimated to be 43.7 MBq/km2 (1.18 mCi/km2) from core samples analyses in 1977, which is almost the same with the reported value of cumulative fallout 239, 240Pu in Tokyo.


A modified azoalbumin technique for the assay of proteolytic enzymes for use in blood group serology.
A spectrophotometric assay of proteolytic enzyme activity is described in which the optical density obtained by a given enzyme preparation is related (2 alpha less than 0.01) to the serological activity of that preparation in a two stage technique. The assay appears to be of use for the standardisation of enzyme preparations for use in blood group serology.


[Type distribution of hyperlipoproteinemias in 645 diabetics].
On 645 diabetics of the treatment-groups diet (n = 180), sulfonylurea therapy (n = 250), biguanide therapy (n = 50), and insulin therapy (n = 165) the concentrations of cholesterol and triglyceride were determined and the lipoprotein electrophoresis was carried out. 29% of the diabetics showed hyperliproteinaemias, males and females in the same frequency. In about 54% the type IV was observed most frequently, then followed the types IIb with 25% and the type IIa with about 13%. The dependence of triglycerides on the metabolic situation and the body weight was proved in diabetics. The cholesterol values were independent on form of therapy, body weight and metabolic situation. In the group with sulfonylurea in about 49%, in the group with insulin in about 37% and in the group with diet and buformin only in about 32% a hyperlipoproteinaemia was proved. The opinion is represented that the high percentage of hyperlipoproteinaemia in diabetics and the connected with this additionally increased risk for vascular complications should lead to the introduction of diagnostics and therapy of hyperlipoproteinaemia into the programme of the diabetes dispensaries of the GDR. For the treatment of diabetes the principle should be taken into consideration that a diabetes is optimally regulated only then, when apart from the blood sugar values also the blood fats are within the normal.


Monitoring secretory membrane with FM1-43 fluorescence.
FM1-43 and similar styryl dyes have proven useful as probes for membrane trafficking because they reversibly stain membranes, are impermeable to membranes, and are more fluorescent when bound to membranes than when in solution. Because these dyes stain membranes in an activity-dependent manner, they are ideal for studies of neurotransmitter release mechanisms such as synaptic vesicle recycling, exocytosis, and endocytosis. FM dyes have been used in conjunction with other techniques such as fluorescent calcium indicator dyes and electrophysiological techniques to elucidate mechanisms of presynaptic calcium homeostasis and modulation of neurotransmitter release. Presynaptic membranes have been marked by FM dyes in studies of synaptogenesis and reinnervation. As a probe for endocytosed membranes, these dyes have been used to examine vacuole formation in yeast. These versatile membrane dyes are useful in a variety of applications.


An easier way of inserting the Shah permavent grommet.
Ventilation tube insertion plays an important role in the management of otitis media with effusion. In selected cases, repeated grommet insertion due to persistent eustachian tube dysfunction necessitates the need for longer-term ventilation. Insertion of such tubes can however occasionally be more difficult than insertion of standard grommets. One such long-term ventilation tube is the Shah permavent grommet. This paper describes a simple modification of the technique that is less time-consuming and less traumatic.


Distribution of gamma-glutamyl transpeptidase in human pancreas: immunohistochemical study with a monoclonal antibody.
We studied in detail the distribution pattern of gamma-glutamyl transpeptidase (gamma-GT) in human pancreas, using the immunoperoxidase technique and a monoclonal antibody to human kidney gamma-GT. Positive reaction was confined exclusively to the luminal surface of the centroacinar cells and the epithelia of the intercalated, intralobular, and interlobular ducts. The immunoreaction was stronger in the intercalated and intralobular ducts than in the interlobular ducts. The acinar cells did not show any reaction. The islets of Langerhans were heavily surrounded by the ducts, and normal islet cells showed no reaction. The course of the ducts, from the acinar lumina to the interlobular ducts, was delineated by using reaction sites positive for gamma-GT as markers. The courses of the ducts, which comprise the pathway for pancreatic juice, were found to vary widely in their connections with each other, especially between the intralobular and interlobular ducts. At least three separate routes could be identified.


Expression of functionally active sialylated human erythropoietin in plants.
Recombinant human erythropoietin (rhEPO), a glycohormone, is one of the leading biopharmaceutical products. The production of rhEPO is currently restricted to mammalian cell expression systems because of rhEPO's highly complex glycosylation pattern, which is a major determinant for drug-efficacy. Here we evaluate the ability of plants to produce different glycoforms of rhEPO. cDNA constructs were delivered to Nicotiana benthamiana (N. benthamiana) and transiently expressed by a viral based expression system. Expression levels up to 85 mg rhEPO/kg fresh leaf material were achieved. Moreover, co-expression of rhEPO with six mammalian genes required for in planta protein sialylation resulted in the synthesis of rhEPO decorated mainly with bisialylated N-glycans (NaNa), the most abundant glycoform of circulating hEPO in patients with anemia. A newly established peptide tag (ELDKWA) fused to hEPO was particularly well-suited for purification of the recombinant hormone based on immunoaffinity. Subsequent lectin chromatography allowed enrichment of exclusively sialylated rhEPO. All plant-derived glycoforms exhibited high biological activity as determined by a cell-based receptor-binding assay. The generation of rhEPO carrying largely homogeneous glycosylation profiles (GnGnXF, GnGn, and NaNa) will facilitate further investigation of functionalities with potential implications for medical applications.


A role for Lyt-2+ T cells in resistance to cutaneous leishmaniasis in immunized mice.
The role of Lyt-2+ T cells in immunologic resistance to cutaneous leishmaniasis was analyzed by comparing infection patterns in resistant C57BL/6 mice and susceptible BALB/c mice induced to heal their infections after sub-lethal irradiation or i.v. immunization, with similar mice treated in vivo with anti-Lyt-2 antibodies. Administration of anti-Lyt-2 mAb resulted in a dramatic reduction in the number of lymphoid cells expressing the Lyt-2+ phenotype. Such treatment led to enhanced disease in both resistant C57BL/6 and irradiated BALB/c mice, as assessed by lesion size, but did not affect the capacity of these mice to ultimately resolve their infections. In contrast, anti-Lyt-2 treatment totally blocked the induction of resistance in i.v. immunized mice. These results suggest, that Lyt-2+ T cells may play a role in immunity to a Leishmania major infection and that their relative importance to resistance may depend on how resistance is induced.


Molecular models of two competitive inhibitors, IL-2delta2 and IL-2delta3, generated by alternative splicing of human interleukin-2.
Molecular models of IL-2delta2 and IL-2delta3, two alternative splice variants of human IL-2 without exon 2 and 3, respectively, are described. These alternative splice variants attract particular interest as potential competitive inhibitors of the cytokine. Tertiary structure of IL-2 consists of four-helix bundle including helices A, B, C and D and a beta-pleated sheet. Exon 2 encodes the A-B loop (Asn30-Lys49 residues) linking helices A and B running in one direction. Rotation of the helix A around putative centre during the construction of IL-2delta2 model have not produced any significant changes in the hydrophobic core of IL-2 molecule. However, a large hole was formed on the surface of IL-2delta2 molecule instead of A-B loop in IL-2 fold. A high affinity IL-2 receptor is formed by combination of alpha, beta, and gamma(c) chains. Comparison of the model of the receptor bound IL-2 with the model of IL-2delta2 has shown that their beta-chain binding sites have minimum differences as distinct from alpha and gamma(c) chain-binding sites. Exon 3 encodes Ala50-Lys97 fragment which forms helices B and C with their short connecting loop. Model IL-2delta3 consists of helices A and D and long linking loop. This loop was composed of A-B and C-D loops which run in opposite directions in IL-2 structure and contain beta-strands making a beta-pleated sheet. Conformation of the linking loop relatively to helices A and D was stabilized by creation of a disulphide bond between cysteines 105 and 125. In addition, the hydrophobic residues of beta-sheet interact with the hydrophobic surface of A-D helical complex and close the latter from contacts with solution. Comparison of the model of IL-2 bound to receptor with IL-2delta3 model has shown that absence of helices B and C in IL-2delta3 model results in insignificant conformational changes only in residues interacting with gamma(c) chain of the receptor. The beta/gamma(c) heterodimer is an intermediate affinity receptor of IL-2. Most likely, both IL-2delta2 and IL-2delta3 are naturally occurring IL-2 antagonists since they keep the ability of binding with an intermediate affinity receptor of this cytokine and fail to engage the alpha chain of its high affinity receptor.


Electronic spectrum of Kekulé patterned graphene considering second neighbor-interactions.
The effects of second-neighbor interactions in Kekulé-Y patterned graphene electronic properties are studied starting from a tight-binding Hamiltonian. Thereafter, a low-energy effective Hamiltonian is obtained by projecting the high energy bands at the Γ point into the subspace defined by the Kekulé wave vector. The spectrum of the low energy Hamiltonian is in excellent agreement with the one obtained from a numerical diagonalization of the full tight-binding Hamiltonian. The main effect of the second-neighbour interaction is that a set of bands gains an effective mass and a shift in energy, thus lifting the degeneracy of the conduction bands at the Dirac point. This band structure is akin to a 'pseudo spin-one Dirac cone', a result expected for honeycomb lattices with a distinction between one third of the atoms in one sublattice. Finally, we present a study of Kekulé patterned graphene nanoribbons. This shows that the previous effects are enhanced as the width decreases. Moreover, edge states become dispersive, as expected due to second neighbors interaction, but here the Kek-Y bond texture results in an hybridization of both edge states. The present study shows the importance of second neighbors in realistic models of Kekulé patterned graphene, specially at surfaces.


Evidence of intrauterine growth restriction and growth hormone deficiency in 49,XXXXY syndrome.
49,XXXXY is an X and Y chromosome variation that occurs in 1:85,000 to 1:100,000 live male births. Previous case studies have described boys with this disorder to be shorter than average when compared with boys with only one extra chromosome and with the mean stature in a small cohort reported to range from the seventh to 33rd percentile. The origin behind the possible differences in height between boys with 47,XXY and 49,XXXXY is currently unknown, however one study hypothesized that it was due to a difference in the expression of the SHOX gene. This study reports on the anthropometric measurements of 84 boys with 49,XXXXY. Forty-five percent of children with 49,XXXXY were found to be below the third percentile in height at the time of evaluation. In addition, 7.14% of the cohort were diagnosed and given treatment for growth hormone deficiency (GHD). The analysis of this cohort demonstrates that the below average heights seen throughout childhood in this population potentially begins prenatally and suggests that boys with 49,XXXXY may be at a higher risk for intrauterine growth restriction (IUGR) and GHD. Future research is needed to investigate the etiology of the poor growth in boys with 49,XXXXY and evaluate the incidence of GHD and IUGR in this population.


Comparative Immunohistochemical Analysis of p53 and Alpha-SMA in Ameloblastoma, AOT and OKC.
Ameloblastoma is a benign but highly infiltrative tumour, a behaviour that is lacking in adenomatoid odontogenic tumour but partly shared by the odontogenic keratocyst which possesses a unique intrinsic growth potential with marked ability for destroying bone and a high tendency recurrence. High frequency of stromal myofibroblasts (assessed with alpha smooth muscle actin (α-SMA) correlates with aggressive behaviour while p53-cell cycle regulation system is critical in odontogenic tumours with immunoreactivity signifying prognostic status. This study aims to determine and compare the immunoreactivity of these selected tumours to p53 and α-SMA in order to establish if a relationship exists between the frequency and pattern of distribution of myofibroblasts and the behaviour of these lesions. 69 blocks of ameloblastoma, and 23 each of adenomatoid odontogenic tumor (AOT), and odontogenic keratocyst (OKC/KCOT) were retrieved. Immunohistochemistry technique was applied for evaluation of these two markers staining with primary antibodies to p53 and -SMA and the frequency and pattern of distribution of myofibroblasts and immunoreactivity to p53 analysed and compared using ANOVA. p was set at <0.05. Immunoreactivity to p53 and α-SMA was highest in ameloblastoma (solid compared to unicystic) with highest mean positive cells to α-SMA (29.7±20.1) and p53 (28.3±24.5) in plexiform ameloblastoma. This suggests that ameloblastoma was the most aggressive of tumours studied. Different pharmacological agents that can regulate stromal MF are useful aids to decrease the need for radical surgery in extensive and aggressive odontogenic tumours.