ATP-dependent proteases controlling mitochondrial function in the yeast Saccharomyces cerevisiae.
Regulated protein degradation by ATP-dependent proteases plays a fundamental role in the biogenesis of mitochondria. Membrane-bound and soluble ATP-dependent proteases have been identified in various subcompartments of this organelle. Subunits composing these proteases are evolutionarily conserved from yeast to humans and, in support of an endosymbiotic origin of mitochondria, evolved from prokaryotic ancestors: the PIM1/Lon protease is active in the matrix of mitochondria, while the i-AAA protease and the m-AAA protease mediate the turnover of inner membrane proteins. Most of the knowledge concerning the biogenesis and the physiological role of ATP-dependent proteases comes from studies in the yeast Saccharomyces cerevisiae. Proteases were found to be required for mitochondrial stasis, for the maintenance of the morphology of the organelle and for mitochondrial genome integrity. ATP-dependent proteolysis is crucial for the expression of mitochondrially encoded subunits of respiratory chain complexes and for the assembly of these complexes. Hence, mitochondrial ATP-dependent proteases exert multiple roles which are essential for the maintenance of cellular respiratory competence.


Effect of dietary protein, alfalfa, and zeolite on excretory patterns of 5',5',7',7'-[3H]zearalenone in rats.
A series of experiments was conducted to determine how dietary protein, alfalfa, or zeolite influence the excretory patterns of zearalenone (Z), a uterotropic mycotoxin synthesized by Fusarium fungi. Rats were fed diets containing 16.3% casein, 40% casein, 11.2% casein + 25% alfalfa, or 25% casein + 25% alfalfa. Also fed were diets containing 0, 1, 2, or 5% anion exchange zeolite. Tracer doses of [3H]Z were administered either as a constituent of the diet or as a topical application on the skin at the base of the skull. When Z was administered orally, no differences were seen in the fraction of the dose excreted in urine or feces as a result of varying dietary levels of alfalfa and protein. Topical doses resulted in rats fed 25% casein + 25% alfalfa or 40% casein excreting more Z in urine than those fed 25% alfalfa or 16.3% casein. Fecal excretion of Z was greatest for rats fed 25% casein + 25% alfalfa whereas rats fed 40% casein excreted more fecal Z than those fed 16.3% casein. Feeding Z to rats receiving dietary zeolite resulted in a positive correlation between dietary zeolite and fecal excretion of Z but a negative correlation with urinary excretion of Z. Topical administration of Z produced a positive correlation between dietary zeolite and fecal Z excretion but no effect on urinary excretion. It may be concluded that protein and alfalfa treatments alleviate Z toxicosis through increased metabolism whereas zeolite binds Z in the digestive tract to prevent absorption.


Uses of guided imagery for pain control by african-american and white women with metastatic breast cancer.
Understanding the meanings patients attach to their experiences can help clinicians and researchers to more adequately address a patient's experience with cancer pain. Indeed, many patients seem to desire to and benefit from attaching meaning to the disease and its treatment. In particular, many patients are drawn to guided imagery as a tool in the management of cancer-related anxiety and pain. Using excerpts from African-American and White women's breast cancer narratives, we show that breast cancer survivors use guided imagery as a vehicle for reconnecting to the self, to make sense of their experiences with breast cancer, and as a tool for managing cancer pain. Cancer pain increases the disruption in the connection between the body and the mind that is already part of the illness experience. Guided imagery can be regarded as one response to this problem, and may be understood as an attempt to reconnect mind and body in a manner that increases the sense of control, thereby alleviating the suffering of the survivor.


Diversity of Prdm9 zinc finger array in wild mice unravels new facets of the evolutionary turnover of this coding minisatellite.
In humans and mice, meiotic recombination events cluster into narrow hotspots whose genomic positions are defined by the PRDM9 protein via its DNA binding domain constituted of an array of zinc fingers (ZnFs). High polymorphism and rapid divergence of the Prdm9 gene ZnF domain appear to involve positive selection at DNA-recognition amino-acid positions, but the nature of the underlying evolutionary pressures remains a puzzle. Here we explore the variability of the Prdm9 ZnF array in wild mice, and uncovered a high allelic diversity of both ZnF copy number and identity with the caracterization of 113 alleles. We analyze features of the diversity of ZnF identity which is mostly due to non-synonymous changes at codons -1, 3 and 6 of each ZnF, corresponding to amino-acids involved in DNA binding. Using methods adapted to the minisatellite structure of the ZnF array, we infer a phylogenetic tree of these alleles. We find the sister species Mus spicilegus and M. macedonicus as well as the three house mouse (Mus musculus) subspecies to be polyphyletic. However some sublineages have expanded independently in Mus musculus musculus and M. m. domesticus, the latter further showing phylogeographic substructure. Compared to random genomic regions and non-coding minisatellites, none of these patterns appears exceptional. In silico prediction of DNA binding sites for each allele, overlap of their alignments to the genome and relative coverage of the different families of interspersed repeated elements suggest a large diversity between PRDM9 variants with a potential for highly divergent distributions of recombination events in the genome with little correlation to evolutionary distance. By compiling PRDM9 ZnF protein sequences in Primates, Muridae and Equids, we find different diversity patterns among the three amino-acids most critical for the DNA-recognition function, suggesting different diversification timescales.


How likely is the sense-B-noise to affect patients with subcutaneous implantable cardioverter-defibrillators and can we solve that problem in every case?
Sense-B-noise is a newly reported possible cause of inappropriate shocks in patients with subcutaneous implantable cardioverter-defibrillators (S-ICDs). The nature of that noise is unknown, it is not related to mechanical failure of the S-ICD system. Reprogramming to the secondary sensing vector is suggested by the producer as a possible solution. We analyzed the medical records of S-ICD recipients from two university clinical centers (Gdansk and Szczecin, Poland). Our aim was to determine the rate of sense-B-noise, and whether the secondary sensing vector would be available for reprogramming if such a problem occurred in our patients. The sense-B-noise issue affected three patients in our cohort (3%), which corresponds to the incidence of 0.012 events per patient-year of follow-up. The primary vector was permanently used in 47 patients (52%), secondary in 28 (31%), and alternate in 16 (17%), respectively. Therefore, the total number of patients potentially vulnerable to sense-B noise (with the primary or alternate vector programmed permanently) was 63 (69%). Among those 63 patients, 51 individuals (81%) had also the secondary vector available for permanent use. The sense-B-noise affected 3% of patients in our cohort, with an incidence of 0.012 per patient-year of follow-up. Most patients potentially vulnerable to sense-B noise could be reprogrammed to the secondary sensing vector, if necessary. Further investigation of the sense-B-noise issue is needed.


[Transport and metabolism of daidzein in Caco-2 cell model].
To investigate the intestinal absorption and metabolism of daidzein in Caco-2 cell model. The damage of daidzein to Caco-2 cell was evaluated by MTT value and alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) activity. With the reference to propranolol, the apparent permeability coefficient (Papp) of daidzein was measured through the monolayer under the different concentrations and pH media. The metabolites were also measured by enzyme hydrolysis. Daidzein had no damage to the growth of the Caco-2 cells in the concentration of 1-50 microg/mL. The apparent permeability coefficient (Papp) of daidzein across the monolayer showed concentration- and pH- independent manner, which was similar to that of transcellcular marker-propranolol, suggesting the good absorption of daidzein in vivo. By hydrolysis with beta-glucuronidase, low metabolites were detected in monolayer and transport medium, verifying the existence of glucuronides and sulfates. The daidzein absorption in Caco-2 cell model is a passive and transcellular transport and quite good in the intestines.


Interactions of normal, dysplastic, and malignant mammary epithelial cells with fibronectin in vivo and in vitro.
Previous studies have demonstrated that normal and malignant mouse mammary cells are indistinguishable in many surface-related properties that often denote transformation of other cell types such as fibroblasts. In the present investigation, the interactions of normal, dysplastic, and malignant mammary epithelial cells with fibronectin in tissues and cultures were examined by indirect immunofluorescence. Cells lining the lumina of ducts and alveoli in normal and dysplastic mouse and human mammary tissues abutted a layer of fibronectin along their basal surfaces that included the region of the basement membrane and the underlying stroma. Moreover, double staining for keratin and fibronectin revealed that myoepithelial cells were surrounded by the matrix protein. In contrast, tumor cells in adenocarcinomas and ductal carcinomas were not directly associated with fibronectin. The accumulation of fibronectin in primary cultures prepared from mouse mammary tissues paralleled the distribution seen in vivo. A matrix of fibronectin formed beneath normal and preneoplastic mammary cells within 4 to 6 days after plating, whereas tumor cells were negative, regardless of the age or density of the culture. This correlation with in vivo results did not extend to cells of established mammary tumor culture lines which readily accumulated pericellular networks of fibronectin. Addition of exogenous fibronectin to primary cultures enhanced formation of a basal matrix by normal cells but had no effect on the negative status of the tumor cells. The results indicate that mammary tumor cells in tissues and in primary cultures have an altered capacity to interact with fibronectin. However, this alteration is not necessarily expressed by established mammary tumor cell lines.


International multicentre validation of the arteriovenous malformation-related intracerebral haemorrhage (AVICH) score.
The recently published arteriovenous malformation-related intracerebral haemorrhage (AVICH) score showed better outcome prediction for patients with arteriovenous malformation (AVM)-related intracerebral haemorrhage (ICH) than other AVM or ICH scores. Here we present the results of a multicentre, external validation of the AVICH score. All participating centres (n=11) provided anonymous data on 325 patients to form the Spetzler-Martin (SM) grade, the supplemented SM (sSM) grade, the ICH score and the AVICH score. Modified Rankin score (mRS) at last follow-up (mean 25.6 months) was dichotomized into favourable (mRS 0-2, n=210) and unfavourable (mRS 3-6;n=115). Univariate and AUROC analyses were performed to validate the AVICH score. Except nidus structure and AVM size, all single parameters forming the SM, sSM, ICH and AVICH score and the scores itself were significantly different between both outcome groups in the univariate analysis. The AVICH score was confirmed to be the highest predictive outcome score with an AUROC of 0.765 compared with 0.705 for the ICH score and 0.682 for the sSM grade. The multicentre-validated AVICH score predicts clinical outcome superior to pre-existing scores. We suggest the routine use of this score for future clinical outcome prediction and in clinical research. NCT02920645.


Upstream versus downstream thrombin inhibition.
For a long time, vitamin K antagonists (VKA) have been the preferred drugs for the anticoagulation management of both atrial fibrillation and venous thromboembolism. Hereby, the major purpose is to attenuate the onset of thrombosis without affecting hemostasis. Areas covered: Nowadays, non-vitamin K anticoagulants (NOAC), a new class of oral anticoagulants is available for the above-mentioned indications. NOAC are at least as effective and safer with regard to intracranial bleedings compared to VKA, but major bleedings still occur. For this reason, the search for safer anticoagulants is still ongoing. Expert commentary: There are several unmet needs in NOAC management, including selection of optimal drug and dose, uncertainty on specific conditions and lack of drug persistence. There remains to be an important need for safer anticoagulants; 'upstream' anticoagulants including inhibitors of factor XIa may provide additional benefit related to fewer bleeding complications.


Use of a mobile cart influenza program for vaccination of hospital employees.
Rates of annual influenza vaccination of healthcare workers (HCWs) remained low in our university hospital. This study was conducted to evaluate the impact of a mobile cart influenza vaccination program on HCW vaccination. From 2000 to 2002, the employee health service continued its annual influenza vaccination program and the mobile cart program was implemented throughout the institution. This program offered influenza vaccination to all employees directly on the units. Each employee completed a questionnaire. Vaccination rates were analyzed using the Mantel-Haenszel test. The program proposed vaccination to 50% to 56% of the employees. Among the nonvaccinated employees, 52% to 53% agreed to be vaccinated. The compliance with vaccination varied from 61% to 77% among physicians and medical students and from 38% to 55% among nurses and other employees. Vaccination of the chief or associate professor of the unit was associated with a higher vaccination rate of the medical staff (P < .01). Altogether, the vaccination program led to an increase in influenza vaccination among employees from 6% in 1998 and 7% in 1999 before the mobile cart program to 32% in 2000, 35% in 2001, and 32% in 2002 (P < .001). The mobile cart program was associated with a significantly increased vaccination acceptance. Our study was able to identify HCW groups for which the mobile cart was effective and highlight the role of the unit head in its success.


First-trimester exposure to amoxycillin/clavulanic acid: a prospective, controlled study.
The number of published studies on the use of amoxycillin/clavulanic acid during pregnancy is small and so is the number of pregnancies investigated in those studies. In this study we wished to investigate prospectively the safety of intrauterine exposure to amoxycillin/clavulanic acid in a relatively large cohort of women. Women treated (n = 191) with amoxycillin/clavulanic acid during the first trimester of pregnancy were recruited from two teratogen information centres in Israel. Exposed women were matched for age, smoking habits and alcohol consumption with 191 controls exposed to amoxycillin only for similar medical indications. Maternal age, birth weight, gestational age at delivery, rates of live births and abortions were comparable between the two groups. Rates of major malformations in the amoxycillin/clavulanic acid group (3/158, 1.9%) did not differ significantly from controls (5/163, 3%) (P = 0.49, relative risk = 0.62, 95% confidence interval 0.15, 2.55), and were within the expected baseline risk for the general population. These data suggest that exposure to amoxycillin/clavulanic acid during pregnancy is unlikely to be associated with an increased risk of malformations.


Potential probiotic Bifidobacterium animalis ssp. lactis 420 prevents weight gain and glucose intolerance in diet-induced obese mice.
Alterations of the gut microbiota and mucosal barrier are linked with metabolic diseases. Our aim was to investigate the potential benefit of the potential probiotic Bifidobacterium animalis ssp. lactis 420 in reducing high-fat diet-induced body weight gain and diabetes in mice. In the obesity model, C57Bl/6J mice were fed a high-fat diet (60 energy %) for 12 weeks, and gavaged daily with B. lactis 420 (109 cfu) or vehicle. In the diabetes model, mice were fed a high-fat, ketogenic diet (72 energy % fat) for 4 weeks, with a 6-week subsequent treatment with B. lactis 420 (108-1010 cfu/day) or vehicle, after which they were analysed for body composition. We also analysed glucose tolerance, plasma lipopolysaccharide and target tissue inflammation using only one of the B. lactis 420 groups (109 cfu/day). Intestinal bacterial translocation and adhesion were analysed in a separate experiment using an Escherichia coli gavage. Body fat mass was increased in both obese (10.7 ± 0.8 g (mean ± standard error of mean) vs. 1.86 ± 0.21 g, P<0.001) and diabetic mice (3.01 ± 0.4 g vs. 1.14 ± 0.15 g, P<0.001) compared to healthy controls. Treatment with B. lactis 420 significantly decreased fat mass in obese (7.83 ± 0.67 g, P=0.007 compared to obese with vehicle) and diabetic mice (1.89 ± 0.16 g, P=0.02 for highest dose). This was reflected as reduced weight gain and improved glucose tolerance. Furthermore, B. lactis 420 decreased plasma lipopolysaccharide levels (P<0.001), liver inflammation (P=0.04), and E. coli adhesion in the distal gut (P<0.05). In conclusion, B. lactis 420 reduces fat mass and glucose intolerance in both obese and diabetic mice. Reduced intestinal mucosal adherence and plasma lipopolysaccharide suggest a mechanism related to reduced translocation of gut microbes.


Consistent re-modeling of signaling pathways and its implementation in the TRANSPATH database.
The data model of the signaling pathways database TRANSPATH has been re-engineered to a three-layer model comprising experimental evidences and summarized pathway information, both in a mechanistically detailed manner, and a "semantic" projection for the abstract overview. Each molecule is described in the context of a certain reaction in the multidimensional space of posttranslational modification, molecular family relationships, and the biological species of its origin. The new model makes the data better suitable for reconstructing signaling pathways and networks and mapping expression data, for instance from microarray experiments, onto regulatory networks.


How to cope with food allergy symptoms?
Food allergy symptoms may involve a wide variety of organs (skin, gastrointestinal tract, and eyes) and systems (respiratory, circulatory, and neurological). They are often associated. Their severity ranges from mild to potentially life-threatening reactions and the presentation from acute to chronic. Food allergies have been the focus of multiple studies, position papers, and guidelines in recent years. They have defined an approach in classifying symptoms as mild to anaphylactic, distinguishing objective from subjective symptoms, and describing their heterogeneity, specific phenotypes or syndromes (e.g., lipid transfer protein syndrome or pollen food syndrome). Cofactors can also influence food allergy reactions. Symptoms of non-IgE-gastrointestinal food allergies, illustrated by eosinophilic esophagitis and food-protein-induced enterocolitis syndrome, are also now better understood and defined. Improvement in the knowledge of food allergy symptoms is crucial for correct diagnosis and a personalized treatment approach. Through a better description and understanding of symptoms, the diversity of food allergies is now better known. The next step is to harmonize symptom assessment not only for clinicians but also for patients, researchers, and public health stakeholders, to describe food allergy phenotypes and their underlying mechanisms and endotypes, to develop targeted management.


Hepatitis C virus-associated hepatocellular carcinoma as a second primary malignancy: exposing an overlooked presentation of liver cancer.
Chronic hepatitis C virus (HCV) infection is one of the leading causes of hepatocellular carcinoma (HCC) worldwide. Antiviral therapy in patients with HCV infection reduces the risk of primary HCC development by 71%-75%. HCV-infected patients with different primary cancers are also at risk for HCC development as a second primary malignancy (HCC-SPM). Limited information is available on the occurrence and characteristics of HCC-SPM. Herein, we determine the prevalence and clinical features of HCV-associated HCC-SPM when compared to primary HCC. Patients with HCV-associated HCC seen at MD Anderson Cancer Center (2011-2017) were enrolled in a prospective observational study. Patients with multiple cancers diagnosed simultaneously or with hepatitis B virus or HIV coinfections were excluded. At enrollment, patients completed a questionnaire on medical history and HCC risk factors. Information on demographics, comorbidities, HCV treatment, tumor characteristics, treatment modalities, and virologic and oncologic outcomes were extracted from the medical records. Among 171 consecutive patients with HCV-associated HCC enrolled, 26 (15%) had HCC-SPM. Most of the underlying primary cancers were solid tumors (85%). In 12 (46%) of these patients, the diagnosis was made incidentally while undergoing surveillance for primary malignancies, and the majority (81%) had their primary cancer in remission. Most patients (72%) with documented HCV viral load had chronic viremia due to lack of diagnosis, lack of treatment, or prior unsuccessful treatment of HCV infection and only 28% had undetectable viral load following successful antiviral therapy. The overall median survival for both groups was 29 months (95% CI: 23-35) without difference between groups (p=0.2). Cancer patients with any malignancies must be screened for HCV as HCC-SPM can develop in 15% of infected patients. Early HCV diagnosis and treatment should be attempted to prevent the development of HCC-SPM, a condition associated with high mortality in cancer survivors.


Expression of the zic1, zic2, zic3, and zic4 genes in early chick embryos.
The zic genes encode a family of transcription factors with important roles during early development. Since little is known about zic gene expression in chick embryos, we have characterized the expression patterns of the zic1, zic2, zic3, and zic4 (zic1-4) genes during neurulation and somitogenesis. We used in situ hybridization to analyze the expression patterns of the zic1-4 genes during early chick development (HH stages 7-19). The zic1-3 genes showed both overlapping and gene-specific expression patterns along the length of the dorsal neural tube and in the dorsal parts of the somites. In addition, unique expression domains of zic genes included: zic2 in the neural plate, periotic mesoderm and limb buds; zic3 in the paraxial mesoderm surrounding the neural plate, in presomitic mesoderm and in the most recently formed epithelial somites; zic2 and zic3 in developing eyes. zic4 expression was limited to dorsal fore- and midbrain regions and, unlike the expression of the zic1-3 genes, zic4 expression was not detected in the hindbrain and trunk. This was in contrast to more extensive zic4 expression in other vertebrates. The zic1-3 genes were expressed in both overlapping and unique domains within the neural tube, somites and other ectoderm and mesoderm-derived structures in the future head and trunk. zic4 expression, however, was limited to dorso-anterior regions of the future brain. This is the first comprehensive study of zic1-4 gene expression in chick embryos during neurulation and somitogenesis.


Diffusion Limitations and Translocation Barriers in Atomically Thin Biomimetic Pores.
Ionic transport in nano- to sub-nano-scale pores is highly dependent on translocation barriers and potential wells. These features in the free-energy landscape are primarily the result of ion dehydration and electrostatic interactions. For pores in atomically thin membranes, such as graphene, other factors come into play. Ion dynamics both inside and outside the geometric volume of the pore can be critical in determining the transport properties of the channel due to several commensurate length scales, such as the effective membrane thickness, radii of the first and the second hydration layers, pore radius, and Debye length. In particular, for biomimetic pores, such as the graphene crown ether we examine here, there are regimes where transport is highly sensitive to the pore size due to the interplay of dehydration and interaction with pore charge. Picometer changes in the size, e.g., due to a minute strain, can lead to a large change in conductance. Outside of these regimes, the small pore size itself gives a large resistance, even when electrostatic factors and dehydration compensate each other to give a relatively flat-e.g., near barrierless-free energy landscape. The permeability, though, can still be large and ions will translocate rapidly after they arrive within the capture radius of the pore. This, in turn, leads to diffusion and drift effects dominating the conductance. The current thus plateaus and becomes effectively independent of pore-free energy characteristics. Measurement of this effect will give an estimate of the magnitude of kinetically limiting features, and experimentally constrain the local electromechanical conditions.


Identification of a chemokine receptor profile characteristic for mediastinal large B-cell lymphoma.
Mediastinal large B-cell lymphomas (MLBCLs) are considered as a subtype of diffuse large B-cell lymphoma; however, they exhibit completely different patterns of dissemination. Since they share a number of surface markers with thymic B cells, a close relationship was assumed. MLBCLs arise extranodally within the anterior mediastinum and have a low propensity to metastasize. To address the preferential positioning of MLBCL, we focused on homeostatic chemokines involved in B-cell compartmental homing in secondary lymphoid organs, which are also capable of shaping lymphoid niches in ectopic sites. Here, we applied immunohistochemistry to assess chemokine receptor and ligand expression in situ. Flow cytometry was used to identify the chemokine receptor profile on an MLBCL-derived cell line, Karpas1106 and on thymic B cells. Migration assays were performed to examine functionality of chemokine receptors. Electrophoretic mobility shift assay was applied to score for NF-kappaB activity. Using immunohistochemistry, we obtained an unexpectedly low-expression frequency for the chemokine receptors CXCR5 and CCR7 in primary lesions. Although the mature B-cell marker CCR6 was absent in most cases, the lineage aberrant marker CCR9 emerged in the majority of MLBCL cases. Given the role of NF-kappaB in the transcriptional activation of CCR7, we identified the involvement of the noncanonical activation pathway in MLBCLs. MLBCLs exhibit a diagnostic chemokine receptor profile that is instrumental in the discrimination from diffuse large B-cell lymphoma not otherwise specified and classical Hodgkin lymphoma. Furthermore, we suggest that low-abundance expression of CCR7 and CXCR5 may hinder lymphoma cells from nodal dissemination.


Elevated CO2 affects anxiety but not a range of other behaviours in juvenile yellowtail kingfish.
Elevated seawater CO2 can cause a range of behavioural impairments in marine fishes. However, most studies to date have been conducted on small benthic species and very little is known about how higher oceanic CO2 levels could affect the behaviour of large pelagic species. Here, we tested the effects of elevated CO2, and where possible the interacting effects of high temperature, on a range of ecologically important behaviours (anxiety, routine activity, behavioural lateralization and visual acuity) in juvenile yellowtail kingfish, Seriola lalandi. Kingfish were reared from the egg stage to 25 days post-hatch in a full factorial design of ambient and elevated CO2 (~500 and ~1000 μatm pCO2) and temperature (21 °C and 25 °C). The effects of elevated CO2 were trait-specific with anxiety the only behaviour significantly affected. Juvenile S. lalandi reared at elevated CO2 spent more time in the dark zone during a standard black-white test, which is indicative of increased anxiety. Exposure to high temperature had no significant effect on any of the behaviours tested. Overall, our results suggest that juvenile S. lalandi are largely behaviourally tolerant to future ocean acidification and warming. Given the ecological and economic importance of large pelagic fish species more studies investigating the effect of future climate change are urgently needed.


MR microscopy of the rat carotid artery after balloon injury by using an implanted imaging coil.
Neointimal hyperplasia after angioplasty was followed in vivo in rats by using MR microscopy and surgically implanted RF imaging coils. By using an inductively coupled pick-up coil, the arteries were imaged 4 days before and 3, 7, and 14 days after angioplasty with a 3DFT spin echo sequence. Eight of 10 angioplastied rats showed moderate to severe stensois based MR measures of lumen diameter reduction from baseline images. There was a good correlation between total wall thickness between MR and hematoxylin and eosin (H&E)-stained sections obtained on the last day. Arteries in the intact and sham groups remained unchanged from baseline measurements. Because this imaging technique examines the artery under in vivo conditions of arterial pressure and flow, it promises to be a useful tool for evaluating pharmacological and mechanical methods of reducing the incidence of vascular stenosis.


Measures of physical activity using cell phones: validation using criterion methods.
Physical activity is associated with reduced risks of many chronic diseases. Data collected on physical activity in large epidemiological studies is often based on paper questionnaires. The validity of these questionnaires is debated, and more effective methods are needed. This study evaluates repeated measures of physical activity level (PAL) and the feasibility of using a Java-based questionnaire downloaded onto cell phones for collection of such data. The data obtained were compared with reference estimates based on the doubly labeled water method and indirect calorimetry (PAL(ref)). Using a Java-based cell phone application, 22 women reported their physical activity based on two short questions answered daily over a 14-day period (PAL(cell)). Results were compared with reference data obtained from the doubly labeled water method and indirect calorimetry (PAL(ref)). Results were also compared against physical activity levels assessed by two regular paper questionnaires completed by women at the end of the 14-day period (PAL(quest1) and PAL(quest2)). PAL(cell), PAL(quest1), and PAL(quest2) were compared with PAL(ref) using the Bland and Altman procedure. The mean difference between PAL(cell) and PAL(ref) was small (0.014) with narrow limits of agreement (2SD = 0.30). Compared with PAL(ref), the mean difference was also small for PAL(quest1) and PAL(quest2) (0.004 and 0.07, respectively); however, the limits of agreement were wider (PAL(quest1), 2SD = 0.50 and PAL(quest2), 2SD = 0.90). The test for trend was statistically significant for PAL(quest1) (slope of regression line = 0.79, P = .04) as well as for PAL(quest2) (slope of regression line = 1.58, P < .001) when compared with PAL(ref). A Java-based physical activity questionnaire administered daily using cell phones produced PAL estimates that agreed well with PAL reference values. Furthermore, the limits of agreement between PAL obtained using cell phones, and reference values were narrower than for corresponding estimates obtained using paper questionnaires. Java-based questionnaires downloaded onto cell phones may be a feasible and cost-effective method of data collection for large-scale prospective studies of physical activity.


Critical insights on the demographic concept of "birth spacing": locating Nef in family well-being, bodies, and relationships in Senegal.
Birth spacing has emerged since the early 1980s as a key concept to improve maternal and child health, triggering interest in birth spacing practices in low-income countries, and drawing attention to prevailing norms in favour of long birth intervals in West Africa. In Senegal, the Wolof concept of Nef, which means having children too closely spaced in time, is morally condemned and connotes a resulting series of negative implications for family well-being. While Nef and "birth spacing" intersect in key ways, including acknowledging the health benefits of longer birth intervals, they are not translations of each other, for each is embedded in distinct broader cultural and political assumptions about social relations. Most notably, proponents of the demographic concept of birth spacing assume that the practice of using contraception after childbearing to postpone births could contribute to "empowering" women socially. In Senegal, by contrast, preventing Nef (or short birth intervals) is also viewed as strengthening family well-being by allowing women to care more fully for their family. This paper draws on policy documents and interviews to explore women's and men's understanding of Nef, and in turn critically reflect on the demographic concept of birth spacing. Our findings reinforce the relevance of the concept of birth spacing to engage with women and men around family planning services in Senegal. Accounts of the Nef taboo in Senegal also show that social norms stigmatising short birth intervals can legitimise constraints faced by women on control of their body.


Trastuzumab treatment in patients with advanced or recurrent endometrial carcinoma overexpressing HER2/neu.
To study the effect of trastuzumab in patients with progressive or recurrent metastatic endometrial carcinoma shown by immunohistochemistry to overexpress the HER2/neu receptor. Disease progression was examined in 2 patients who met the study criteria, had c-erbB2 gene amplification by fluorescence in situ hybridization, and were treated with trastuzumab following radiation treatment and/or salvage chemotherapy. The clinical responses to trastuzumab as a single agent or in combination with chemotherapy were confirmed in both patients by serial CT scans and serum CA-125 evaluations. These patients with progressive or recurrent metastatic disease experienced relief from their symptoms and prolonged survival with no significant toxicity observed. Trastuzumab may be a viable therapeutic option as single agent or in combination with chemotherapy in patients with advanced, recurrent, and/or metastatic endometrial carcinomas overexpressing HER2/neu.


Spatial stochastic simulation offers potential as a quantitative method for pest risk analysis.
Pest risk analysis represents an emerging field of risk analysis that evaluates the potential risks of the introduction and establishment of plant pests into a new geographic location and then assesses the management options to reduce those potential risks. Development of new and adapted methodology is required to answer questions concerning pest risk analysis of exotic plant pests. This research describes a new method for predicting the potential establishment and spread of a plant pest into new areas using a case study, Ralstonia solanacearum, a bacterial disease of potato. This method combines current quantitative methodologies, stochastic simulation, and geographic information systems with knowledge of pest biology and environmental data to derive new information about pest establishment potential in a geographical region where a pest had not been introduced. This proposed method extends an existing methodology for matching pest characteristics with environmental conditions by modeling and simulating dissemination behavior of a pest organism. Issues related to integrating spatial variables into risk analysis models are further discussed in this article.


Overexpression of the microRNA miR-433 promotes resistance to paclitaxel through the induction of cellular senescence in ovarian cancer cells.
Annually, ovarian cancer (OC) affects 240,000 women worldwide and is the most lethal gynecological malignancy. High-grade serous OC (HGSOC) is the most common and aggressive OC subtype, characterized by widespread genome changes and chromosomal instability and is consequently poorly responsive to chemotherapy treatment. The objective of this study was to investigate the role of the microRNA miR-433 in the cellular response of OC cells to paclitaxel treatment. We show that stable miR-433 expression in A2780 OC cells results in the induction of cellular senescence demonstrated by morphological changes, downregulation of phosphorylated retinoblastoma (p-Rb), and an increase in β-galactosidase activity. Furthermore, in silico analysis identified four possible miR-433 target genes associated with cellular senescence: cyclin-dependent kinase 6 (CDK6), MAPK14, E2F3, and CDKN2A. Mechanistically, we demonstrate that downregulation of p-Rb is attributable to a miR-433-dependent downregulation of CDK6, establishing it as a novel miR-433 associated gene. Interestingly, we show that high miR-433 expressing cells release miR-433 into the growth media via exosomes which in turn can induce a senescence bystander effect. Furthermore, in relation to a chemotherapeutic response, quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed that only PEO1 and PEO4 OC cells with the highest miR-433 expression survive paclitaxel treatment. Our data highlight how the aberrant expression of miR-433 can adversely affect intracellular signaling to mediate chemoresistance in OC cells by driving cellular senescence.


ADP stimulates IP3 formation in human platelets.
Aspirinated human platelets labeled with 32PO4 showed a 1.7-fold increase in [32P]IP3 when stimulated with ADP. ADP-stimulated mobilization of internal Ca2+ and phosphorylation of myosin were enhanced in the presence of extracellular Ca2+ but the increase in IP3 was not significantly affected by external Ca2+. The Ca2+ ionophore, ionomycin, elevated internal Ca2+ and induced myosin phosphorylation without a detectable change in IP3. These results indicate that the mechanism of ADP stimulation of human platelets is similar to that of other platelet agonists and supports the theory that IP3 functions to liberate internal Ca2+.


Projected shifts in coral size structure in the Anthropocene.
Changes in the size structure of coral populations have major consequences for population dynamics and community function, yet many coral reef monitoring projects do not record this critical feature. Consequently, our understanding of current and future trajectories in coral size structure, and the demographic processes underlying these changes, is still emerging. Here, we provide a conceptual summary of the benefits to be gained from more comprehensive attention to the size of coral colonies in reef monitoring projects, and we support our argument through the use of case-history examples and a simplified ecological model. We neither seek to review the available empirical data, or to rigorously explore causes and implications of changes in coral size, we seek to reveal the advantages to modifying ongoing programs to embrace the information inherent in changing coral colony size. Within this framework, we evaluate and forecast the mechanics and implications of changes in the population structure of corals that are transitioning from high to low abundance, and from large to small colonies, sometimes without striking effects on planar coral cover. Using two coral reef locations that have been sampled for coral size, we use demographic data to underscore the limitations of coral cover in understanding the causes and consequences of long-term declining coral size, and abundance. A stage-structured matrix model is used to evaluate the demographic causes of declining coral colony size and abundance, particularly with respect to the risks of extinction. The model revealed differential effects of mortality, growth and fecundity on coral size distributions. It also suggested that colony rarity and declining colony size in association with partial tissue mortality and chronic declines in fecundity, can lead to a demographic bottleneck with the potential to prolong the existence of coral populations when they are characterized by mostly very small colonies. Such bottlenecks could have ecological importance if they can delay extinction and provide time for human intervention to alleviate the environmental degradation driving reductions in coral abundance.


Hepatitis B knowledge, testing, and vaccination among Chinese and Vietnamese adults in Australia.
Hepatitis B is a significant public health challenge within some subpopulations in Australia, including Chinese and Vietnamese migrants. There has been limited research on hepatitis B knowledge and actions in these communities. The authors conducted a self-administered survey among 442 Chinese and 433 Vietnamese in Brisbane. Generally, the knowledge is best described as "moderate." One in 2 could not identify the sexual transmission risk and less than one third knew that sharing foods or drinks did not spread the disease. The majority of Vietnamese (80%) and 60% of Chinese respondents indicated prior testing. Vaccination was reported in 60% of the Vietnamese and in 52% of the Chinese. Knowledge was better among Chinese people who had been tested and vaccinated compared with those who were nontested and nonvaccinated. Only 3.5% of the Chinese, but 11.6% of the Vietnamese, indicated having a positive test result hepatitis B virus. This study helps identify strategies for programs targeting both communities and practitioners.


Does Intraindividual Variability of Personality States Improve Perspective Taking? An Ecological Approach Integrating Personality and Social Cognition.
Research integrating cognitive abilities and personality has focused on the role of personality traits. We propose a theory on the role of intraindividual variability of personality states (hereafter state variability) on perspective taking, in particular, the ability to infer other peoples' mental states. First, we review the relevant research on personality psychology and social cognition. Second, we propose two complementary routes by which state variability relates to anchoring and adjustment in perspective taking. The first route, termed ego-dispersion, suggests that an increased state variability decreases egocentric bias, which reduces anchoring. The second route, termed perspective-pooling, suggests that an increased state variability facilitates efficient adjustment. We also discuss how our theory can be investigated empirically. The theory is rooted in an ecological interpretation of personality and social cognition, and flags new ways for integrating these fields of research.


A Recently Assembled Degradation Pathway for 2,3-Dichloronitrobenzene in Diaphorobacter sp. Strain JS3051.
Diaphorobacter sp. strain JS3051 utilizes 2,3-dichloronitrobenzene (23DCNB), a toxic anthropogenic compound, as the sole carbon, nitrogen, and energy source for growth, but the metabolic pathway and its origins are unknown. Here, we establish that a gene cluster (dcb), encoding a Nag-like dioxygenase, is responsible for the initial oxidation of the 23DCNB molecule. The 2,3-dichloronitrobenzene dioxygenase system (DcbAaAbAcAd) catalyzes conversion of 23DCNB to 3,4-dichlorocatechol (34DCC). Site-directed mutagenesis studies indicated that residue 204 of DcbAc is crucial for the substrate specificity of 23DCNB dioxygenase. The presence of glutamic acid at position 204 of 23DCNB dioxygenase is unique among Nag-like dioxygenases. Genetic, biochemical, and structural evidence indicate that the 23DCNB dioxygenase is more closely related to 2-nitrotoluene dioxygenase from Acidovorax sp. strain JS42 than to the 34DCNB dioxygenase from Diaphorobacter sp. strain JS3050, which was isolated from the same site as strain JS3051. A gene cluster (dcc) encoding the enzymes for 34DCC catabolism, homologous to a clc operon in Pseudomonas knackmussii strain B13, is also on the chromosome at a distance of 2.5 Mb from the dcb genes. Heterologously expressed DccA catalyzed ring cleavage of 34DCC with high affinity and catalytic efficiency. This work not only establishes the molecular mechanism for 23DCNB mineralization, but also enhances the understanding of the recent evolution of the catabolic pathways for nitroarenes. IMPORTANCE Because anthropogenic nitroaromatic compounds have entered the biosphere relatively recently, exploration of the recently evolved catabolic pathways can provide clues for adaptive evolutionary mechanisms in bacteria. The concept that nitroarene dioxygenases shared a common ancestor with naphthalene dioxygenase is well established. But their phylogeny and how they evolved in response to novel nitroaromatic compounds are largely unknown. Elucidation of the molecular basis for 23DCNB degradation revealed that the catabolic pathways of two DCNB isomers in different isolates from the same site were derived from different recent origins. Integrating structural models of catalytic subunits and enzymatic activities data provided new insight about how recently modified enzymes were selected depending on the structure of new substrates. This study enhances understanding and prediction of adaptive evolution of catabolic pathways in bacteria in response to new chemicals.


Tertiary hyperparathyroidism resistant to cinacalcet treatment.
Cinacalcet hydrochloride (cinacalcet) has been reported to be efficacious for patients with tertiary hyperparathyroidism (THPT). We experienced five patients with THPT requiring parathyroidectomy (PTx) because of resistance to cinacalcet treatment and investigated their clinical characteristics and clinical course. The maximum diameter of the parathyroid gland estimated by ultrasonography before renal transplantation was evaluated. Serum total calcium, phosphorus, intact parathyroid hormone (iPTH), alkaline phosphatase (ALP), and creatinine (Cr) levels were investigated every three months after the administration of cinacalcet and at PTx. After surgery, the Cr levels were followed. In all five patients, at least one parathyroid gland had a largest diameter of more than 1 cm, and the mean diameter was 18.7 mm (range 14.9-24.1 mm). Intact PTH and ALP levels gradually increased after the initiation of cinacalcet and the Cr levels transiently increased after PTx. These findings suggest that the existence of a severely enlarged nodular hyperplastic gland is a main factor involved in resistance to cinacalcet.


The cholesterol-dependent cytolysin signature motif: a critical element in the allosteric pathway that couples membrane binding to pore assembly.
The cholesterol-dependent cytolysins (CDCs) constitute a family of pore-forming toxins that contribute to the pathogenesis of a large number of Gram-positive bacterial pathogens.The most highly conserved region in the primary structure of the CDCs is the signature undecapeptide sequence (ECTGLAWEWWR). The CDC pore forming mechanism is highly sensitive to changes in its structure, yet its contribution to the molecular mechanism of the CDCs has remained enigmatic. Using a combination of fluorescence spectroscopic methods we provide evidence that shows the undecapeptide motif of the archetype CDC, perfringolysin O (PFO), is a key structural element in the allosteric coupling of the cholesterol-mediated membrane binding in domain 4 (D4) to distal structural changes in domain 3 (D3) that are required for the formation of the oligomeric pore complex. Loss of the undecapeptide function prevents all measurable D3 structural transitions, the intermolecular interaction of membrane bound monomers and the assembly of the oligomeric pore complex. We further show that this pathway does not exist in intermedilysin (ILY), a CDC that exhibits a divergent undecapeptide and that has evolved to use human CD59 rather than cholesterol as its receptor. These studies show for the first time that the undecapeptide of the cholesterol-binding CDCs forms a critical element of the allosteric pathway that controls the assembly of the pore complex.


Defying gravity and fear: the prevention of falls in community-dwelling older adults.
In older adults, falls represent a very common and dangerous problem. Older adults lack the normal overlap or redundancy of cognitive, sensory, and musculoskeletal mechanisms that help younger people avoid falling while remaining active. Many of the risk factors for falls in older adults are modifiable and deserve aggressive attention. Successful prevention of falls, fall-related injuries, and fear of falling is complicated by the multifactorial nature of the problem. Advanced practice nurses' comprehensive health assessment skills, holistic orientation, and willingness to collaborate with healthcare providers from a range of disciplines are helpful in dealing with the complexity of balance problems and falls. Nurse practitioners can be effective case managers for older adults at risk for falling. Older adults need accurate information, and sometimes counseling, to realize that falls, fall-related injuries, and fear of falls are warning signs that can provide direction and motivation for change and prevention.


The Thermoplasma acidophilum LplA-LplB complex defines a new class of bipartite lipoate-protein ligases.
Lipoic acid is a covalently bound cofactor found throughout the domains of life that is required for aerobic metabolism of 2-oxoacids and for C(1) metabolism. Utilization of exogenous lipoate is catalyzed by a ligation reaction that proceeds via a lipoyl-adenylate intermediate to attach the cofactor to the epsilon-amino group of a conserved lysine residue of protein lipoyl domains. The lipoyl ligases of demonstrated function have a large N-terminal catalytic domain and a small C-terminal accessory domain. Half of the members of the LplA family detected in silico have only the large catalytic domain. Two x-ray structures of the Thermoplasma acidophilum LplA structure have been reported, although the protein was reported to lack ligase activity. McManus et al. (McManus, E., Luisi, B. F., and Perham, R. N. (2006) J. Mol. Biol. 356, 625-637) hypothesized that the product of an adjacent gene was also required for ligase activity. We have shown this to be the case and have named the second protein, LplB. We found that complementation of Escherichia coli strains lacking lipoate ligase with T. acidophilum LplA was possible only when LplB was also present. LplA had no detectable ligase activity in vitro in the absence of LplB. Moreover LplA and LplB were shown to interact and were purified as a heterodimer. LplB was required for lipoyl-adenylate formation but was not required for transfer of the lipoyl moiety of lipoyl-adenylate to acceptor proteins. Surveys of sequenced genomes show that most lipoyl ligases of the kingdom Archaea are heterodimeric. We propose that the presence of an accessory domain provides a diagnostic to distinguish lipoyl ligase homologues from other members of the lipoate/biotin attachment enzyme family.


An electromyographic study of weight-bearing at the elbow joint.
The purpose of this study was to investigate the roles of specific muscles in supporting the elbow joint in full extension during a static weighted situation. Various forearm positions from pronation through supination were used to determine carrying efficiency by monitoring muscle activity. Fine-wire electrodes were inserted into the biceps brachii, brachialis, brachioradialis, and medial head of the triceps in order to record the action potentials of these muscles. The forearm was studied in four different positions: (1) prone, (2) relaxed or natural, (3) semiprone, and (4) supine. These positions were monitored by an elgon potentiometer. The loads for the subjects were from 2-20 kilograms. Reliability coefficients of repeated trials were significantly positive. It was concluded that the muscles played an important role in supporting the elbow in full extension during a detracting force and that there was no significant difference between the muscular activity of males and females. Of the four positions studied, the supine position required the most activity suggesting an inefficient carrying position. The triceps was significantly more active when the arm was in the prone position. The roles of muscle groups in supporting the fully extended elbow joint during a detracting force changed as the forearm moved between supination and pronation. The elbow flexors were most active in the supine position, while the elbow extensor muscle was most active in the prone position.


Deletion of CPEB1 Gene: A Rare but Recurrent Cause of Premature Ovarian Insufficiency.
Premature ovarian insufficiency (POI) may be secondary to chemotherapy, radiotherapy, or environmental factors. Genetic causes are identified in 20-25% of cases, but most POI cases remain idiopathic. This study aimed to identify new genes involved in POI and to characterize the implication of CPEB1 gene in POI. This was a case report and cohort study replicate conducted in academic medical centers. A deletion including CPEB1 gene was first identified in a patient with primary amenorrhea. Secondly, 191 sporadic POI cases and 68 familial POI cases were included. For each patient, karyotype was normal and FMR1 premutation was excluded. Search for CPEB1 deletions was performed by quantitative multiplex PCR of short fluorescent fragments or DNA microarray analysis. Gene sequencing of CPEB1 was performed for 95 patients. We identified three patients carrying a microdeletion in band 15q25.2. The proximal breakpoint, for the three patients, falls within a low-copy repeat region disrupting the CPEB1 gene, which represents a strong candidate gene for POI as it is known to be implicated in oocyte meiosis. No mutation was identified by sequencing CPEB1 gene. Therefore, heterozygous deletion of CPEB1 gene leading to haploinsufficiency could be responsible for POI in humans. Microdeletions of CPEB1 were identified in 1.3% of patients with POI, whereas no mutation was identified. This microdeletion is rare but recurrent as it is mediated by nonallelic homologous recombination due to the existence of low-copy repeats in the region. This result demonstrates the importance of DNA microarray analysis in etiological evaluation and counseling of patients with POI.


Immunotherapy in Breast Cancer- Paving New Roads?
Significant advances have been made in our understanding of the mechanisms of innate and adaptive immunity and the mechanisms by which tumors escape from the normal process of immune surveillance. However, effectively harnessing the power of the immune system to specifically target the tumor cells has proven quite difficult, in large part due to the fact that tumors are composed of patient's own cells, expressing antigens recognized as "self" and thus escaping the immune surveillance. For breast cancer treatment, the first successful inroads in immunotherapy came with the introduction of monoclonal antibodies against a membrane receptor for the epidermal growth factor receptor family (HER2). In 1998 the first monoclonal antibody against the Her 2 antigen was approved for clinical use (trastuzumab). Clinical trials combining trastuzumab with various chemotherapy regimens have consistently shown improvement in survival in both metastatic as well as early stage breast cancer. Another promising approach has been the development of anti HER 2 vaccines. More recently the discovery of immune modulating antibodies (checkpoint inhibitors) and their success in the treatment of cancers such as melanoma and renal cell carcinoma has lead to renewed interest in immune therapies in cancer in general and new clinical trials exploring the role of immune therapies in breast cancer.


Secondary structure prediction for aligned RNA sequences.
Most functional RNA molecules have characteristic secondary structures that are highly conserved in evolution. Here we present a method for computing the consensus structure of a set aligned RNA sequences taking into account both thermodynamic stability and sequence covariation. Comparison with phylogenetic structures of rRNAs shows that a reliability of prediction of more than 80% is achieved for only five related sequences. As an application we show that the Early Noduline mRNA contains significant secondary structure that is supported by sequence covariation.


mRNA translation is not a prerequisite for small interfering RNA-mediated mRNA cleavage.
RNA interference constitutes a major means of eliminating mRNAs, yet how the small interfering RNAs (siRNA) within the RNA-induced silencing complex (RISC) finds its homologous target in the cell remains unknown. An attractive hypothesis is that RNA interference is linked to translation which allows RISC ready access to every translated mRNA. To test whether translation could direct siRNAs to mRNAs, chemical and biological inhibitors of translation and their effects on mRNA cleavage were tested. Our results show that mRNA degradation by siRNAs is not dependent on mRNA translation.


Promoting access to innovation for frail old persons. IAGG (International Association of Gerontology and Geriatrics), WHO (World Health Organization) and SFGG (Société Française de Gériatrie et de Gérontologie) Workshop--Athens January 20-21, 2012.
Frailty tends to be considered as a major risk for adverse outcomes in older persons, but some important aspects remain matter of debate. The purpose of this paper is to present expert's positions on the main aspects of the frailty syndrome in the older persons. Workshop organized by International Association of Gerontology and Geriatrics (IAGG), World Health Organization (WHO) and Société Française de Gériatrie et de Gérontologie (SFGG). Frailty is widely recognized as an important risk factor for adverse health outcomes in older persons. This can be of particular value in evaluating non-disabled older persons with chronic diseases but today no operational definition has been established. Nutritional status, mobility, activity, strength, endurance, cognition, and mood have been proposed as markers of frailty. Another approach calculates a multidimensional score ranging from "very fit" to "severely frail", but it is difficult to apply into the medical practice. Frailty appears to be secondary to multiple conditions using multiple pathways leading to a vulnerability to a stressor. Biological (inflammation, loss of hormones), clinical (sarcopenia, osteoporosis etc.), as well as social factors (isolation, financial situation) are involved in the vulnerability process. In clinical practice, detection of frailty is of major interest in oncology because of the high prevalence of cancer in older persons and the bad tolerance of the drug therapies. Presence of frailty should also be taken into account in the definition of the cardiovascular risks in the older population. The experts of the workshop have listed the points reached an agreement and those must to be a priority for improving understanding and use of frailty syndrome in practice. Frailty in older adults is a syndrome corresponding to a vulnerability to a stressor. Diagnostic tools have been developed but none can integrate at the same time the large spectrum of factors and the simplicity asked by the clinical practice. An agreement with an international common definition is necessary to develop screening and to reduce the morbidity in older persons.


Migraine and drug absorption.
The majority of migraine attacks are associated with gastrointestinal symptoms which add considerably to the distress and inconvenience caused by the headache. When salicylate absorption from effervescent aspirin tablets was studied during migraine, the rate of absorption was found to be reduced relative to that found in non-migrainous volunteers and in the same patients when headache-free. There is evidence that this reduced rate of absorption is caused by gastrointestinal stasis and reduced rate of gastric emptying. Patients in whom aspirin absorption was delayed were more likely to take longer to respond and to require additional treatment. Metoclopramide, which increases gastric emptying rate, has been shown to improve the rate of absorption of aspirin during migraine and also increase the rate of recovery from the attack and avoid the need for additional treatment; effects which were not shown by thiethylperazine. It is likely that delayed absorption during migraine affects some drugs other than aspirin, such as ergotamine, and it is therefore recommended that the most rapidly absorbable formulation should be used. If such treatment is ineffective, metoclopramide may be a useful addition and should be tried before resorting to other routes of administration.


[Early-stage breast cancer - strategies for adjuvant systemic therapy].
Systemic adjuvant treatment is an integral component in the treatment of women with early-stage breast cancer and contributes significantly to the reduction of breast cancer-specific mortality. Decisions regarding adjuvant treatment modalities are becoming increasingly complex and demand individualised risk assessments based not only on clinicopathological criteria such as tumour size and axillary lymph node status but also on molecular predictors of the biological behaviour of the cancer. In addition, with the development of targeted therapies targeting HER-2 and with the availability of third generation aromatase inhibitors, new therapeutic options are available and need to be integrated into the treatment algorithms. In this review, we discuss the guidelines and recommendations concerning adjuvant systemic treatment of early-stage breast cancer based on current risk classifications.


Lactobacillus reuteri reduces bone loss in older women with low bone mineral density: a randomized, placebo-controlled, double-blind, clinical trial.
The importance of the gut microbiome for bone metabolism in mice has recently been demonstrated, but no studies are available in humans. Lactobacillus reuteri ATCCPTA 6475 (L. reuteri 6475) has been reported to increase bone mineral density (BMD) in mice but its effect on the human skeleton is unknown. The objective of this trial was to investigate if L. reuteri 6475 affects bone loss in older women with low BMD. In this double-blind, placebo-controlled study, women from the population who were 75 to 80 years old and had low BMD were randomized to orally receive 1010 colony-forming units of L. reuteri 6475 daily or placebo. The predefined primary end-point was relative change after 12 months in tibia total volumetric BMD (vBMD). Ninety women were included and 70 completed the study. L. reuteri 6475 reduced loss of total vBMD compared to placebo both in the intention-to-treat (ITT) analysis [-0.83% (95% confidence interval [CI], -1.47 to -0.19%) vs. -1.85% (95% CI, -2.64 to -1.07%); mean difference 1.02% (95% CI, 0.02-2.03)] and per protocol analysis [-0.93% (95% CI, -1.45 to -0.40) vs. -1.86% (95% CI, -2.35 to -1.36); mean difference 0.93% (95% CI, 0.21-1.65)]. In general, similar but smaller effects were observed in the secondary bone variable outcomes, but these differences did not reach statistical significance in the ITT population. Adverse events did not differ between groups. Supplementation with L. reuteri 6475 should be further explored as a novel approach to prevent age-associated bone loss and osteoporosis.


Urotensin II inhibited the proliferation of cardiac side population cells in mice during pressure overload by JNK-LRP6 signalling.
Cardiac side population cells (CSPs) are promising cell resource for the regeneration in diseased heart as intrinsic cardiac stem cells. However, the relative low ratio of CSPs in the heart limited the ability of CSPs to repair heart and improve cardiac function effectively under pathophysiological condition. Which factors limiting the proliferation of CSPs in diseased heart are unclear. Here, we show that urotensin II (UII) regulates the proliferation of CSPs by c-Jun N-terminal kinase (JNK) and low density lipoprotein receptor-related protein 6 (LRP6) signalling during pressure overload. Pressure overload greatly upregulated UII level in plasma, UII receptor (UT) antagonist, urantide, promoted CSPs proliferation and improved cardiac dysfunction during chronic pressure overload. In cultured CSPs subjected to mechanical stretch (MS), UII significantly inhibited the proliferation by UT. Nanofluidic proteomic immunoassay showed that it is the JNK activation, but not the extracellular signal-regulated kinase signalling, that involved in the UII-inhibited- proliferation of CSPs during pressure overload. Further analysis in vitro indicated UII-induced-phospho-JNK regulates phosphorylation of LRP6 in cultured CSPs after MS, which is important in the inhibitory effect of UII on the CSPs during pressure overload. In conclusion, UII inhibited the proliferation of CSPs by JNK/LRP6 signalling during pressure overload. Pharmacological inhibition of UII promotes CSPs proliferation in mice, offering a possible therapeutic approach for cardiac failure induced by pressure overload.


Temporal shifts in traits of Vibrio cholerae strains isolated from hospitalized patients in Calcutta: a 3-year (1993 to 1995) analysis.
This study presents results of a surveillance on cholera conducted with hospitalized patients admitted to the Infectious Diseases Hospital, Calcutta, India, from January 1993 to December 1995. The O139 serogroup of Vibrio cholerae dominated in 1993 but was replaced by O1 as the dominant serogroup in 1994 and 1995. The isolation rate of V. cholerae non-O1 non-O139 did not exceed 4.9% throughout the study period, while the isolation rate of the O139 serogroup in 1994 and 1995 was below 9%. No temporal clustering of any non-O1 non-O139 serogroup was observed. With the exception of 1 strain, none of the 64 strains belonging to the non-O1 non-O139 serogroup hybridized with ctx, zot, and ace gene probes, while 97.3 and 97.7% of the O139 and O1 strains, respectively, hybridized with all the three probes. Multiplex PCR studies revealed that all the O1 strains belonged to the EIT or biotype. There was a progressive increase in the cytotoxic response on CHO and HeLa cells evoked by culture supernatants of strains of V. cholerae non-O1 non-O139 isolated during 1994 and 1995 compared with the response evoked by those isolated in 1993. Dramatic shifts in patterns of resistance to antibiotics between strains of V. cholerae belonging to different serogroups and within strains of a serogroup isolated during different time periods were observed. There was a discernible increase in the incidence of multidrug-resistant strains of V. cholerae O1 isolated in 1994 and 1995 compared with that in 1993. On the basis of the results of this study, we predict the possibility of newer variants of V. cholerae emerging in the future.


Mineral resources, environmental issues, and land use.
Contrary to predictions from the 1950s through the mid-1980s, persistent shortages of nonfuel minerals have not occurred, despite prodigious consumption, and world reserves have increased. Global availability of raw materials is relevant to policy decisions regarding mineral development and land use. Justification for environmental protection may exceed that for mining a specific ore body. Demand for environmental accountability is rising worldwide, and new technologies are enabling internalization of costs. Mineral-rich developing nations plagued by inefficient state-owned mining enterprises, high population growth rates, and environmental degradation could realize substantial benefit by reforming government policies to encourage foreign investment in resources and by appropriate allocation of mineral rents.


[Patient-centered medicine for tuberculosis medical services].
The 2011 edition of Specific Guiding Principles for Tuberculosis Prevention calls for a streamlined medical services system capable of providing medical care that is customized to the patient's needs. The new 21st Century Japanese version of the Directly Observed Treatment Short Course (DOTS) expands the indication of DOTS to all tuberculosis (TB) patients in need of treatment. Hospital DOTS consists of comprehensive, patient-centered support provided by a DOTS care team. For DOTS in the field, health care providers should select optimal administration support based on patient profiles and local circumstances. In accordance with medical fee revisions for 2012, basic inpatient fees have been raised and new standards for TB hospitals have been established, the result of efforts made by the Japanese Society for Tuberculosis and other associated groups. It is important that the medical care system be improved so that patients can actively engage themselves as a member of the team, for the ultimate goal of practicing patient-centered medicine. We have organized this symposium to explore the best ways for practicing patient-centered medicine in treating TB. It is our sincere hope that this symposium will lead to improved medical treatment for TB patients. 1. Providing patient-centered TB service via utilization of collaborative care pathway: Akiko MATSUOKA (Hiroshima Prefectural Tobu Public Health Center) We have been using two types of collaborative care pathway as one of the means of providing patient-centered TB services since 2008. The first is the clinical pathway, which is mainly used by TB specialist doctors to communicate with local practitioners on future treatment plan (e.g. medication and treatment duration) of patients. The clinical pathway was first piloted in Onomichi district and its use was later expanded to the whole of Hiroshima prefecture. The second is the regional care pathway, which is used to share treatment progress, test results and other necessary patient information among the relevant parties. The regional care pathway was developed by the Tobu Public Health Center. It is currently being used by several other public health centers in Hiroshima. Utilization of these two pathways has resulted in improved adherence, treatment being offered at local clinics, shorter hospitalization and better treatment outcomes. 2. Patient-centered DOTS in Funabashi-city: Akiko UOZUMI (Funabashi-city Public Health Center) In Funabashi-city, all TB patients, including those with LTBI, are treated under DOTS which recognizes and tries to accommodate the various different needs of each individual patient. For example, various types of DOTS are offered, such as pharmacy-based DOTS and DOTS supported by caregivers of nursing homes. This enables public health nurses to take into consideration both the results of risk assessment and convenience for the patient, and choose DOTS which most effectively support the patient. Furthermore, DOTS in principle is offered face-to-face, so that DOTS providers may not only build relationship of trust with the patient, but also to collect and analyze the necessary information regarding the patient and respond timely when problems arise. Such effort has directly contributed to improved default and treatment rate. 3. Hospital DOTS and clinical path for the treatment of tuberculosis: Kentaro SAKASHITA, Akira FUJITA (Tokyo Metropolitan Tama Medical Center) We introduced a version of hospital DOTS at Tama Medical Center (formerly Fuchu Hospital) in 2004. As part of this three-stage version, patients are allowed to progress to the next stage if they meet the step-up criteria. Following the introduction of this hospital DOTS, the occurrence of drug administration-related incidents decreased and support for patient adherence became easier for health care workers than before. In 2006, we developed a clinical path based on this hospital DOTS with consistent eligibility criteria for patients. This clinical path helped increase the efficiency of medical services in the TB ward. In conclusion, a patient's initiative for tuberculosis treatment can be supported through our hospital's TB treatment system. 4. Survey of TB patients' understanding and satisfaction of hospital DOTS: Yoko NAGATA, Minako URAKAWA, Noriko KOBAYASHI, Seiya KATO (Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association) We surveyed the satisfaction and understanding of recently discharged TB patients regarding DOTS to analyze how to better implement DOTS. The questionnaire consisted of nine items covering knowledge of TB, comfort in talking to and asking questions of the medical staff, explanations given to family members, and motivation for continuing medication. Two hundred and eight of the 228 patients who accepted the questionnaire responded (response rate: 91.2%). The level of understanding and satisfaction tended to be higher among patients in hospitals that employed a primary nursing system, more coverage and duration of DOT, and audiovisual materials for patient education. The level of understanding and satisfaction also tended to be slightly higher among institutions that conducted in-hospital conferences and collaborated with public health centers more frequently. 5. Medical cooperative system against tuberculosis elimination: Dai YOSHIZAWA (Tuberculosis and Infectious disease control division, Ministry of Health, Labour and Welfare) There are 3 points we should consider. First, despite one of the intermediate burden countries, emphasis for infectious incidence is insufficient. Besides new incidence decreases gradually, increased ratio of the elderly causes necessity of implementation against each complications. The second is how find infectious one, especially from high burden countries, before they spread it. Final, unspecific symptoms suffer the patients and medical staff. It's the key of implementation that spread of tuberculosis must be caused by delayed diagnosis.


Whole genome sequence of Vibrio cholerae directly from dried spotted filter paper.
Global estimates for cholera annually approximate 4 million cases worldwide with 95,000 deaths. Recent outbreaks, including Haiti and Yemen, are reminders that cholera is still a global health concern. Cholera outbreaks can rapidly induce high death tolls by overwhelming the capacity of health facilities, especially in remote areas or areas of civil unrest. Recent studies demonstrated that stool specimens preserved on filter paper facilitate molecular analysis of Vibrio cholerae in resource limited settings. Specimens preserved in a rapid, low-cost, safe and sustainable manner for sequencing provides previously unavailable data about circulating cholera strains. This may ultimately contribute new information to shape public policy response on cholera control and elimination. Whole genome sequencing (WGS) recovered close to a complete sequence of the V. cholerae O1 genome with satisfactory genome coverage from stool specimens enriched in alkaline peptone water (APW) and V. cholerae culture isolates, both spotted on filter paper. The minimum concentration of V. cholerae DNA sufficient to produce quality genomic information was 0.02 ng/μL. The genomic data confirmed the presence or absence of genes of epidemiological interest, including cholera toxin and pilus loci. WGS identified a variety of diarrheal pathogens from APW-enriched specimen spotted filter paper, highlighting the potential for this technique to explore the gut microbiome, potentially identifying co-infections, which may impact the severity of disease. WGS demonstrated that these specimens fit within the current global cholera phylogenetic tree, identifying the strains as the 7th pandemic El Tor. WGS results allowed for mapping of short reads from APW-enriched specimen and culture isolate spotted filter papers. This provided valuable molecular epidemiological sequence information on V. cholerae strains from remote, low-resource settings. These results identified the presence of co-infecting pathogens while providing rare insight into the specific V. cholerae strains causing outbreaks in cholera-endemic areas.


Rapid fluctuations in rat barrel cortex plasticity.
Neuronal populations in the sensory cortex exhibit fluctuations in excitability, and the present experiments tested the hypothesis that these variations coincide with peaks and troughs in cortical modifiability. The activity of multiunit neuronal clusters under light urethane anesthesia was recorded through 100-microelectrode arrays implanted in the infragranular layers of rat barrel cortex. Spontaneous activity was characterized by "bursts" of spikes, synchronized across the barrel cortex. This allowed activity at one selected electrode to be taken as a reliable monitor of widespread cortical bursts. We used spikes at the selected electrode to trigger stimulation of two pairs of whiskers during a 50 min conditioning procedure: (1) for the "burst-conditioned" whisker pair, each stimulus was delivered 1 msec after the triggering spike, activating cortex coincident with the burst; and (2) for the "interburst-conditioned" whisker pair, each stimulus was delivered 300 msec after the triggering spike, activating cortex during the trough between bursts. The cross-correlation between cortical neurons in the pairs of columns matching the stimulated whisker pairs was estimated after the termination of the conditioning procedure. Conditioning produced a twofold increase in the degree of co-firing between infragranular neurons in columns receiving burst-conditioned costimulation but no significant change in connectivity between infragranular neurons in columns receiving interburst-conditioned costimulation, although the two pairs of columns received an equal number of sensory inputs. These findings suggest that the strength of co-activity between columns in the barrel cortex can be modified by sensory input patterns during discrete, intermittent intervals time-locked to bursts.


simpleISM-A straight forward guide to upgrade from confocal to ISM.
Resolution in a confocal laser scanning microscopes (CLSM) can be improved if the pinhole is closed. But closing the pinhole will deteriorate the signal to noise ratio (SNR). A simple technique to improve the SNR while keeping the resolution same by upgrading the system to an image scanning microscope. In this paper, we explain in detail, based on an Olympus Fluoview 300 system, how a scanning microscope can be upgraded into an image scanning microscope (ISM) using a simple camera-based detector and an Arduino Due providing a galvo driving and camera synchronization signals. We could confirm a resolution improvement as well as superconcentration and made the interesting observation of a reduced influence of laser fluctuations.


Sinogenic intracranial empyema in children.
Sinogenic intracranial empyema (SIE) is an uncommon complication of sinusitis that can lead to devastating neurologic sequelae. Early recognition of the clinical findings is critical so that proper management can be instituted. To describe the symptoms, signs, and laboratory and imaging findings from one of the largest pediatric SIE case series reported. Descriptive study of a retrospective cohort of all children admitted to Primary Children's Medical Center with SIE between June 2000 and February 2004. Children were identified by a computerized search of Primary Children's Medical Center medical records using the terms "sinusitis" and "brain/subdural/epidural empyema." Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) values of children with SIE were compared with a group of children with uncomplicated sinusitis cared for in the same health care system as outpatients. The medical records of the uncomplicated sinusitis group were not reviewed for any clinical or radiographic data other than CRP and ESR values. Twelve children with SIE were identified. The median age of children with SIE was 11.5 years. Symptoms were usually present 10 days (median) before diagnosis and included headache (10), fever (11), nausea/vomiting (7), mental-status changes (5), and seizures (3). Physical findings included abnormal neurologic examination (9), Pott's puffy tumor (4), and orbital cellulitis (3). Using the Intermountain Health Care system's computerized database, 142 children with uncomplicated sinusitis treated as outpatients were identified. Children with SIE had markedly higher CRP levels (median: 10.05 mg/dL) and ESRs (median: 87 mm/hour) than those with uncomplicated sinusitis (median CRP: 0.7 mg/dL; median ESR: 6 mm/hour). Four children had hyperglycemia. Four children had a lumbar puncture at presentation, and the findings were normal for all of them. Craniofacial imaging included computed tomography (CT) and magnetic resonance imaging (MRI). SIE was not detected in 4 patients who had nonenhanced CT. Axial imaging alone was unable to demonstrate SIE in 1 child with sphenoid and ethmoid sinusitis, and coronal images were needed to demonstrate its presence and extent. The initial facial/orbital imaging studies in 2 patients with physical signs of complicated sinusitis (orbital cellulitis and Pott's puffy tumor) were not adequate to detect SIE. Using contrast-enhanced head CT or MRI, SIE was diagnosed in all 12 children. Children with sinusitis and any neurologic finding, signs of complicated sinusitis such as Pott's puffy tumor or orbital cellulitis, or persistent headache, fever, or nausea and vomiting after antibiotic therapy should have additional evaluation for SIE. Children with hyperglycemia or diabetes may be at increased risk for SIE. The ESR and CRP levels are markedly elevated in children with SIE and may be useful screening tools. MRI with gadolinium is the preferred method to diagnose SIE. If MRI is unavailable, a contrast-enhanced head CT with axial and coronal planes should be obtained. Nonenhanced CT alone lacks sensitivity, and a normal study may be falsely reassuring.


Na⁺/H⁺ exchanger regulatory factor 3 is critical for multidrug resistance protein 4-mediated drug efflux in the kidney.
Na(+)/H(+) exchanger regulatory factor 3 (NHERF3) is a PSD-95/discs large/ZO-1 (PDZ)-based adaptor protein that regulates several membrane-transporting proteins in epithelia. However, the in vivo physiologic role of NHERF3 in transepithelial transport remains poorly understood. Multidrug resistance protein 4 (MRP4) is an ATP binding cassette transporter that mediates the efflux of organic molecules, such as nucleoside analogs, in the gastrointestinal and renal epithelia. Here, we report that Nherf3 knockout (Nherf3(-/-)) mice exhibit profound reductions in Mrp4 expression and Mrp4-mediated drug transport in the kidney. A search for the binding partners of the COOH-terminal PDZ binding motif of MRP4 among several epithelial PDZ proteins indicated that MRP4 associated most strongly with NHERF3. When expressed in HEK293 cells, NHERF3 increased membrane expression of MRP4 by reducing internalization of cell surface MRP4 and consequently, augmented MRP4-mediated efflux of adefovir, a nucleoside-based antiviral agent and well known substrate of MRP4. Examination of wild-type and Nherf3(-/-) mice revealed that Nherf3 is most abundantly expressed in the kidney and has a prominent role in modulating Mrp4 levels. Deletion of Nherf3 in mice caused a profound reduction in Mrp4 expression at the apical membrane of renal proximal tubules and evoked a significant increase in the plasma and kidney concentrations of adefovir, with a corresponding decrease in the systemic clearance of this drug. These results suggest that NHERF3 is a key regulator of organic transport in the kidney, particularly MRP4-mediated clearance of drug molecules.


Serum Levels of Th2-Type Immunoglobulins are Increased in Weanling Mice Subjected to Acute Wasting Protein-Energy Malnutrition.
The bulk of our knowledge on immunosuppression in malnutrition comes from the experiments done on cell-mediated immunity. However, malnutrition-induced modifications of humoral immunity have been less understood. The objective of this study was to determine the effects of acute protein-energy malnutrition on serum levels of immunoglobulins and their subclasses in murine models. Male and female C57BL/6J mice were allocated to one of the four groups: (1) zero-time control (ZC) (19 days of age); (2) ad libitum intake of a complete purified diet (control group [CG]); (3) restricted intake of the complete diet (restricted group [RG]); and ad libitum intake of an isocaloric low-protein diet (low-protein group [LP]). The three groups other than the zero-time control were maintained on their respective regimens for 14 days, i.e. from 19 through 33 days of age. The restricted intake protocol produced weight loss through energy deficiency (marasmic-type malnutrition), whereas the low-protein diet caused wasting through inadequate protein nitrogen and induced a condition mimicking incipient kwashiorkor. Though serum levels of IgG1 and IgE (Th2-type immunoglobulins) in RG and LP mice were significantly higher than those in CG mice, serum levels of IgG2a and IgG3 (Th1-type immunoglobulins) did not show any significant difference between those three groups. Interestingly, serum levels of IgG2b (another Th2-type immunoglobulin) in LP mice were significantly higher than those in CG and RG mice. In ZC mice serum levels of total IgG, IgG1, IgG2b and IgG3 were significantly higher than those in the other three groups. We concluded that during acute malnutrition, Th1/Th2 balance is apparently shifted towards Th2 arm. This deviation seems to be more prominent during acute protein deficiency. The increased serum levels of immunoglobulins in ZC mice were probably due to the intestinal uptake of those proteins from maternal milk.


Effectiveness comparison of multiple sticky-trap configurations for sampling Pseudacteon spp. phorid flies (Diptera: Phoridae).
A variety of traps have been developed for monitoring introduced populations of Pseudacteon spp. phorid flies (Diptera: Phoridae) across their established range in the United States. Such traps typically exploit common aspects of phorid fly biology and behavior, such as their attraction to live or dead red imported fire ants, Solenopsis invicta Buren (Hymenoptera: Formicidae), as well as the perching behavior of these parasitoids. However, populations of multiple species of phorid flies have been established in the United States to serve as biological control agents against S. invicta, and it is unclear if all trap designs are equally effective in sampling this variety of phorid species. This study investigated the effectiveness of six trap designs simultaneously during three sampling events in south-central Texas. Interactions between two species of phorid flies (Pseudacteon tricuspis Borgmeier and P. curvatus B.) and their hosts have been intensively studied at this location for over eight years. When analyzed independently, there were no significant differences in the mean number of P. curvatus or P. tricuspis phorids collected by any of the trap designs during any of the sampling events. However, when the total number of phorids collected were combined, significant trap performance differentials were observed during the October 2010 sampling event. Furthermore, there were significant differences among male flies during the September 2012 observation. Additionally, a trap component cost comparison is provided. The consistent and relatively equivalent performance of the phorid traps investigated in these trials suggests that all are appropriate for phorid surveillance, and cost and ease-of-use considerations may be the most important criteria when selecting a trap design.


Macrocyclic cell penetrating peptides: a study of structure-penetration properties.
Arginine-rich cell penetrating peptides are short cationic peptides able to cross biological membranes despite their peptidic character. In order to optimize their penetration properties and further elucidate their mechanisms of cellular entry, these peptides have been intensively studied for the last two decades. Although several parameters are simultaneously involved in the internalization mechanism, recent studies suggest that structural modifications influence cellular internalization. Particularly, backbone rigidification, including macrocyclization, was found to enhance proteolytic stability and cellular uptake. In the present work, we describe the synthesis of macrocyclic arginine-rich cell penetrating peptides and study their cellular uptake properties using a combination of flow cytometry and confocal microscopy. By varying ring size, site of cyclization, and stereochemistry of the arginine residues, we studied their structure-uptake relationship and showed that the mode and site of cyclization as well as the stereochemistry influence cellular uptake. This study led to the identification of a hepta-arginine macrocycle as efficient as its linear nona-arginine congener to enter cells.


Cancer cells release anaphylatoxin C5a from C5 by serine protease to enhance invasiveness.
Anaphylatoxin C5a indirectly fosters cancer cells through recruitment of myeloid-derived suppressor cells (MDS) for inhibiting antitumor CD8+ T cells and induction of neovascularization. We recently found activation of cancer cells by C5a directly via the C5a-receptor (C5aR; CD88) to enhance invasiveness. Thus, C5a possibly contributes to cancer progression rather than elimination. C5a generation in cancer tissues has been reported; however, the mechanism is not fully elucidated. Cancer cell expression of complement regulatory molecules suggests inefficient C5a generation through activation of the complement system in response to cancer cells. To explore another C5a generation mechanism in cancer tissues, we examined cancer cells for C5a-releasing activity from C5. C5a was present in C5-supplemented culture media of cancer cells including C5aR-expressing cells, and the media enhanced C5aR-expressing cancer cell invasion, which was abolished by anti-C5a antibody. The C5a-releasing activity was absent in the supernatants of the media and was inhibited by aprotinin, a serine protease inhibitor, and decanoyl-Arg-Val-Lys-Arg-chloromethylketone but not by inhibitors specific for cysteine, acid, or metal proteases. These results indicated C5a release from C5 by a cancer cell membrane-bound serine protease that can cleave peptide bonds at the carboxy-terminal site of paired basic amino acid residues. Cancer cell C5a release from the complement-immobilized plasma supported feasibility of this cancer cell protease-dependent C5a generation in cancer tissues. The new mechanism of C5a generation suggests self-activation of C5aR-expressing cancer cells to enhance invasiveness and induction of MDS recruitment and neovascularization to create a microenvironment favorable for cancer progression.


Use of bovine mesenteric vein in rescue vascular access surgery.
We describe a technique for rescue surgery of autologous arterovenous fistula (AVF), using bovine mesenteric vein (BMV), which may be used in patients with autologous AVF malfunction caused by steno-occlusion on the arterial side or by fibrosis of the first portion of the vein. To preserve the autologous AVF, we replaced the diseased portion of the artery, or the first centimeters of the vein, by a segment of BMV, with the aim of saving the patency and functionality of the access. We used this technique in 16 cases. All patients underwent hemodialysis treatment immediately after the procedure. Infection or aneurismal dilatation of the graft in implanted BMV was never observed.


Donor cells' contribution to osteogenesis in experimental cancellous bone grafts.
Osteogenesis in subcutaneous, cancellous bone isografts in the rat was assessed at 16 days from the uptake of 85Sr. by comparing the uptake in grafts devoid of one or more cellular components, the contribution of each to osteogenesis was estimated. The per cent contribution of each of the osteogenetic elements was: endosteal lining cells together with marrow stroma, 60%; periosteal cells, 30%; free, hemopoietic marrow cells and osteocytes, no significant contribution. These findings are compared with those previously reported for cortical bone isografts and it appears that the rate of osteogenesis is 2.2 times higher in cancellous grafts than in cortical grafts of the same weight, but only 1.7 times higher when grafts of the same volume are compared. Osteogenetic and hemopoietic activities associated with marrow appear to arise from different groups of cells. It is shown how, with iliac bone marrow, these two groups of cells may be separated physically, the hemopoietic cells being removed by means of a jet of saline, and the osteogenetic cells by subsequent enzyme treatment plus a second jet of saline. The osteogenetic and hemopoietic elements were difficult to separate from bone marrow within the femoral diaphysis.


Ectopic Hormonal Syndromes: Cushing's Syndrome and Acromegaly.
Ectopic hormone production may be present in neuroendocrine and non-endocrine neoplasms. Ectopic sources of growth hormone, adrenocorticotropin (ACTH), or their releasing factors are uncommon but clinically relevant. Ectopic ACTH tumors have been studied more than the rest, but there are still no comprehensive multidisciplinary guidelines that include all the pitfalls in the diagnosis and management of ectopic hormonal syndromes and the neoplasms associated with ectopic Cushing or acromegaly. The frequency of neuroendocrine neoplasms and other neoplasms with neuroendocrine differentiation has been increasing in recent decades. The review of the available data on these tumors, their classification, and improvements in diagnostic and therapeutic procedures is important to understand the relevance of ectopic Cushing's syndrome and acromegaly in clinical practice.


Breast cancer recurrence dynamics following adjuvant CMF is consistent with tumor dormancy and mastectomy-driven acceleration of the metastatic process.
The aim of this study was to better understand human breast cancer biology by studying how the timing of metastasis following primary resection is affected by adjuvant CMF (cyclophoshamide, methotrexate, 5-fluorouracil) chemotherapy. Discrete hazards of recurrence and recurrence risk reductions for treated patients relative to controls were analyzed for all patients enrolled in two separate randomized clinical trials [study 1 (386 women): no further treatment versus 12 cycles of CMF; study 2 (459 women): six versus 12 cycles of CMF] and a historical group (396 women: surgery alone) of axillary node-positive patients undergoing mastectomy. (i) Nearly all CMF benefit occurs during the first 4 years following resection/chemotherapy. (ii) The CMF recurrence rate reduction is largely restricted to two specific spans. These temporally separate recurrence clusters occur during the first and third year of follow-up, while the second-year recurrences are weakly affected. (iii) Prolonging adjuvant treatment from 6 to 12 months partially alters this recurrence timing, without appreciably affecting the overall recurrence rate. (iv) These effects upon the dynamics of post-resection occurrence are menopausal status-independent. At least two different therapeutically vulnerable proliferative events, resulting in clinical appearance of two metastasis temporally distinct clusters of post-resection cancer recurrence, apparently occur during the administration of adjuvant chemotherapy. Metastases that transpire outside of these temporal windows are refractory to adjuvant therapy. The dynamics of both post-treatment recurrence risk and CMF effectiveness are similar for both pre- and postmenopausal women, suggesting that post-resection mechanisms by which chemotherapy prevents metastases are similar, but of different magnitude in pre- and postmenopausal women. These findings are consistent with a metastasis model that includes tumor dormancy in specific micrometastatic phases (single cells and avascular foci) and with the acceleration of the metastatic process by the surgical resection of the primary breast cancer.


Anomalous pancreaticobiliary junction (APBJ) with the drainage of the uncinate process into the minor papilla: demonstration by MRI.
We report a case of a 30-year-old patient with anomalous pancreaticobiliary junction (APBJ) that has not been described before. The patient had a clinical history of recurrent attacks of pancreatitis, cholangitis and cholecystitis that were confirmed by abnormal laboratory values. Endoscopic retrograde cholangiopancreatography revealed a 20 mm long junction of choledoc and pancreatic duct, and uncinate process draining into the minor papilla. On MR cholangiopancreatography, strictures at the junction of hepatic duct, increased gallbladder wall thickness and intraductal stone in the pancreatic duct were demonstrated as complementary findings. Other MRI findings included decreased signal intensity of the pancreas consistent with fibrosis from past pancreatitis attacks and atrophy of the left liver lobe.