[A case of alpha-fetoprotein producing primary intracranial embryonal carcinoma treated with combination chemotherapy with cis-platinum, vinblastine and bleomycin (author's transl)].
A case of alpha-fetoprotein (AFP) producing primary intracranial embryonal carcinoma was reported with special reference to the chemotherapy. The patient was a 14-year-old male who had suffered from vomiting and disturbance of consciousness. CT scan revealed a tumor originating in the anterior part of the third ventricle and expanding into both lateral ventricles. Right frontotemporal craniotomy was performed and the tumor was totally removed under the microscope. The histological diagnosis was embryonal carcinoma. Inspite of the elevated amount of AFP in the serum, we could not verify the yolk sac element in the surgical specimen. Three months later, he became drowsy and another CT scan revealed recurrence of the tumor. Ommaya's reservoir was placed and CSF was drained to control the intracranial hypertension. But the disturbance of consciousness did not improve. We then started a combination chemotherapy with cis-platinum, vinblastine and bleomycin. Cis-platinum was given in a dosage of 20 mg/m2 body surface area as a 15 min. intravenous infusion for 5 consecutive days every 3 weeks for three courses. Vinblastine was given in a dosage of 0.4 mg/kg body weight intravenously for 2 consecutive days every 3 weeks for three courses. Bleomycin was given in a dosage of 30 mg intravenously 6 hours after vinblastine weekly for a total of 12 weeks. The AFP level of the serum and CSF was monitored every several days. After the chemotherapy, the AFP level of the serum and CSF decreased. Repeated CT scan revealed no evidence of tumor. His clinical condition improved remarkably. Toxicity was vomiting, proteinuria and leukopenia, but not so severe. Proteinuria continued after the chemotherapy, but BUN and creatinine did not elevate. It was emphasized that the combination chemotherapy with cis-platinum, vinblastine and bleomycin is effective remission-induction treatment for AFP producing primary intracranial embryonal carcinoma.


Epinephrine-induced cyclic AMP accumulation in the psoriatic epidermis.
Although there are many reports concerning different beta-adrenergic responsiveness in involved and uninvolved skin of psoriasis, previous experiments have been done mainly by using keratome-sliced skin, which contains unknown amounts of stratum corneum, dermis, skin appendages, etc. In order to determine the effect of epinephrine on the cyclic AMP level of 'pure' epidermis in psoriasis, a microdissection technique was employed. Basal levels of cyclic AMP in the involved epidermis were slightly higher than in the uninvolved epidermis (involved 1.9 +/- 0.3 pmoles/mg dry weight; uninvolved 1.3 +/- 0.3 pmoles/mg d.w.). This difference was not statistically significant (p greater than 0.1). The response to epinephrine by the involved epidermis (8.4 +/- 1.0 pmoles/mg d.w.) was much lower than that in the uninvolved epidermis (23.3 +/- 4.3 pmoles/mg d.w.). The difference was statistically highly significant (p less than 0.005). Our data show that psoriatic involved epidermis per se had a reduced beta-adrenergic responsiveness, which might be significantly involved in the pathophysiology of psoriasis.


Does mRNA structure contain genetic information for regulating co-translational protein folding?
Currently many facets of genetic information are ill-defined. In particular, how protein folding is genetically regulated has been a long-standing issue for genetics and protein biology. And a generic mechanistic model with supports of genomic data is still lacking. Recent technological advances have enabled much needed genome-wide experiments. While putting the effect of codon optimality on debate, these studies have supplied mounting evidence suggesting a role of mRNA structure in the regulation of protein folding by modulating translational elongation rate. In conjunctions with previous theories, this mechanistic model of protein folding guided by mRNA structure shall expand our understandings of genetic information and offer new insights into various biomedical puzzles.


The structure of calcium-ammonia solutions by neutron diffraction.
The microscopic structures of calcium-ammonia solutions have been established by using neutron diffraction. Total structure factors measured at 230 K reveal immediately the evolution of an uncommonly intense diffraction prepeak in the metallic solutions. As concentration is increased from 4 mole percent metal to 10 mole percent metal (i.e., saturation), this feature intensifies and shifts from 0.6 to 0.9 A(-1). It is therefore evidence of well developed intermediate-range ordering among the solvated cations, and is a microstructural signature of the observed strong phase separation of metallic (concentrated) and nonmetallic (dilute) solutions. The technique of isotopic labelling of *N by 15N was then used in conjunction with difference analysis to focus on the solvent structure in metallic solutions at 4 and 10 mole percent metal. These nitrogen-centered functions are analyzed in conjunction with classical Monte Carlo computer simulation techniques, to provide us with detailed insight into the calcium solvation and the extent of hydrogen bonding. We find that calcium is solvated by approximately 6-7 ammonia molecules, with a Ca-N distance of around 2.45 A. There is evidence of hydrogen bonding among the solvent molecules, even in the saturated 10 mole percent metal solution.


Protocatechuic acid inhibits oleic acid-induced vascular smooth muscle cell proliferation through activation of AMP-activated protein kinase and cell cycle arrest in G0/G1 phase.
Protocatechuic acid (PCA) has been implicated in the progression of atherosclerosis. The proliferation of vascular smooth muscle cells (VSMC) may play a crucial role in the pathogenesis of atherosclerosis. Adenosine 5′-monophosphate-activated protein kinase (AMPK) additionally exerts several beneficial effects on vascular function and improves vascular abnormalities. The current study sought to determine whether PCA has an inhibitory effect on VSMC proliferation under oleic acid (OA) treatment. A7r5 cells were treated with OA (150 μM) or cotreated with OA and PCA (150 μg/mL) for 24 and 48 h. PCA-treated cells were found to cause an increase in G0/G1 cell cycle arrest. Western blotting showed that PCA increased the expressions of p53 and p21Cip1, subsequently decreasing the expression of cyclin E1 and Cdk2. In addition, PCA induced phosphorylation of AMPK and inhibited the expression of fatty acid synthase, Akt-p, and Skp2 after stimulation with OA. After treatment with AMPK inhibitor, the effects of PCA mentioned above were reversed. Taken together, PCA inhibited OA-induced VSMC proliferation through AMPK activation and down-regulation of FAS and AKT signals, which then blocks G0/G1 phase cell cycle progression. These findings provide a new insight into the protective properties of PCA on VSMC, which may constitute a novel effective antiatherosclerosis agent.


Periodic fever syndromes--a clinical overview.
Most hereditary periodic fever syndromes known today have their onset in the first year of life. Only two, namely Familial Mediterranean Fever (FMF) and TNF-Receptor Associated Periodic Syndrome (TRAPS) occur later, with most patients having become symptomatic by their twentieth birthday. Therefore this review will concentrate on FMF and TRAPS, the latter being a very rare disease, while the former has become somewhat more common in mid-Europe as a result of migration.


Expression and Prognostic Value Identification of Methylenetetrahydrofolate Dehydrogenase 2 (MTHFD2) in Brain Low-Grade Glioma.
This study aimed to reveal the potential function of methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) and emphasized its importance in brain low-grade glioma (LGG). We firstly explored the differential expression of MTHFD2 mRNA in LGG and normal tissues, followed by correlation analysis of MTHFD2 mRNA expression with patient's clinical characteristics. MTHFD2 protein expression in LGG and subcellular location were also evaluated. Then, survival analysis was performed to reveal the influence of MTHFD2 expression on the overall survival of patients, and Cox regression analysis was applied to predict the prognostic factor for overall survival of LGG. Finally, we performed functional analysis to reveal potential MTHFD2-associated pathways involved in LGG. We found that MTHFD2 was highly expressed in LGG patients (P<0.05), and MTHFD2 expression was related to patient's age and IDH mutation status (all P<0.05). MTHFD2 protein was mainly localized to the mitochondria. Survival analysis showed that high expression of MTHFD2 desirably improved the prognosis of LGG patients (P<0.001), especially for those patients with age ≥45 years (P<0.05). But independent prognostic role of MTHFD2 in LGG was not observed. Pathway enrichment analysis indicated that MTHFD2 high expression significantly and positively participated in the pathway of one carbon pool by folate (all P<0.05). High expression of MTHFD2 was observed in LGG, which was favorable for the overall survival of LGG patients. Our results assumed that MTHFD2 high expression might play a pivotal role in LGG through positively regulating pathway of one carbon pool by folate.


Inhibition of Proliferation and Expression of N-ras in Hepatoma Cells by Antioxidation Treatment.
Kunming mice inoculated with hepatoma cell (H22) suspension subcutaneously at their right axilla were administered orally with antioxidants such as vitamine E, beta-carotene, glutamine, kappa-selenocarrageenan and polysaccharide-peptide of coriolus (PSP) solution. It was found that the inoculated hepatoma growth was suppressed to various extents. The two kinds of polysaccharide antioxidants improved non-specific immunity, enhanced the nitrogen monoxide (NO) content in plasma and strengthened the inhibition of hepatoma. Above antioxidants added in the culture of 7721 human hepatoma cells inhibited the cell proliferation and inducedits apoptosis. Meanwhile, the activity of glutathione peroxidase (GSH-Px) in the plasma of mice increased and the content of malondialdehyde (MDA) decreased. H(2)O(2) in low concentration improved the cancer cell proliferation and inhanced the expression of Mn-SOD c-fos and c-jun, but led to cells apoptosis or necrosis in high concentration. The mechanism of antioxidants inhibiting tumor growth and improving cancer cells apoptosis might be that, on the one hand, the antioxidants blocked the free radicals signal transduction on cancer cells proliferation, and on the other hand, they improved the release of NO through enhancing the non-specific immunity, so inhibiting the cancer cells proliferation directly.


[The protective efficacy of rabbit endogenous nitric oxide against acute rabbit lung ischemia-reperfusion injury and its mechanism].
To investigate the protective efficacy of rabbit endogenous nitric oxide (NO) against acute rabbit lung injury associated with ischemia-reperfusion and explore the possible mechanism. The rabbit lung ischemia-reperfusion (LIR) model was established; thirty-two adult New Zealand white rabbits were randomly divided into four groups. The rabbits of control group underwent sham operation. In group LIR, the rabbits' left lung hili were clamped for 60 minutes and then released. In group LIR+L-Arg (L-arginine), the rabbits were operated upon as those in group LIR, but L-Arg (200 mg/kg) was infused into blood circulation as substrate for NO generation before removal of the clip. In group LIR+L-NNA (L-ng-nitro-Arginine), the rabbits passed through the same operation as in group LIR, but L-NNA (10 mg/kg) was infused into circulation as an inhibitor against NO generation before reperfusion. After reperfusion for 60 minutes, the lung tissues were harvested for histological examination, and the wet to dry ratio of lung tissue weight (W/D), myeloperoxidase (MPO) activity, malondialdehyde (MDA) content as well as the ratio of nitrate/nitrite (N/N) were measured respectively. The group LIR had greater lung tissue W/D, higher MPO activity and MDA content, lower ratio of N/N, and serious pulmonary edema as compared with group ShO (P<0.01). But in group LIR+L-Arg, the degree of pulmonary edema was alleviated, the MPO activity and MDA content were decreased, and the ratio of N/N increased; there was statistically significant difference between group LIR and group LIR+Arg in respect to the above indices (P<0.01). However, in group LIR+L-NNA, the pulmonary edema was even more severe, the MPO activity and MDA content were significantly higher those that in group LIR or group LIR+L-Arg (P<0.01). The endogenous release of pulmonary NO can attenuate the acute lung injury associated with LIR, and the mechanisms may involve the protective efficacy conferred by endogenous NO against accumulation of neutrophil in lung, against pulmonary microvascular permeability, and against the oxygen free radical injury to lung.


Diastolic field stimulation: the role of shock duration in epicardial activation and propagation.
Detailed knowledge of tissue response to both systolic and diastolic shock is critical for understanding defibrillation. Diastolic field stimulation has been much less studied than systolic stimulation, particularly regarding transient virtual anodes. Here we investigated high-voltage-induced polarization and activation patterns in response to strong diastolic shocks of various durations and of both polarities, and tested the hypothesis that the activation versus shock duration curve contains a local minimum for moderate shock durations, and it grows for short and long durations. We found that 0.1-0.2-ms shocks produced slow and heterogeneous activation. During 0.8-1 ms shocks, the activation was very fast and homogeneous. Further shock extension to 8 ms delayed activation from 1.55 ± 0.27 ms and 1.63 ± 0.21 ms at 0.8 ms shock to 2.32 ± 0.41 ms and 2.37 ± 0.3 ms (N = 7) for normal and opposite polarities, respectively. The traces from hyperpolarized regions during 3-8 ms shocks exhibited four different phases: beginning negative polarization, fast depolarization, slow depolarization, and after-shock increase in upstroke velocity. Thus, the shocks of >3 ms in duration created strong hyperpolarization associated with significant delay (P < 0.05) in activation compared with moderate shocks of 0.8 and 1 ms. This effect appears as a dip in the activation-versus-shock-duration curve.


Collective electronic excitations in the ultra violet regime in 2-D and 1-D carbon nanostructures achieved by the addition of foreign atoms.
Plasmons in the visible/UV energy regime have attracted great attention, especially in nano-materials, with regards to applications in opto-electronics and light harvesting; tailored enhancement of such plasmons is of particular interest for prospects in nano-plasmonics. This work demonstrates that it is possible, by adequate doping, to create excitations in the visible/UV regime in nano-carbon materials, i.e., carbon nanotubes and graphene, with choice of suitable ad-atoms and dopants, which are introduced directly into the lattice by low energy ion implantation or added via deposition by evaporation. Investigations as to whether these excitations are of collective nature, i.e., have plasmonic character, are carried out via DFT calculations and experiment-based extraction of the dielectric function. They give evidence of collective excitation behaviour for a number of the introduced impurity species, including K, Ag, B, N, and Pd. It is furthermore demonstrated that such excitations can be concentrated at nano-features, e.g., along nano-holes in graphene through metal atoms adhering to the edges of these holes.


Knee registries: state of the art.
Sports injuries, trauma and the globally ageing and obese population require increasing levels of knee surgery. Shared decision making has replaced the paternalistic approach to patient management. Evidence-based medicine underpins surgical treatment strategies, from consenting an individual patient to national healthcare system design. The evolution of successful knee-related registries starting from specific arthroplasty registries has given rise to ligament reconstruction, osteotomy and cartilage surgery registries developing as platforms for surgical outcome data collection. Stakeholders include surgeons and their patients, researchers, healthcare systems, as well as the funding insurers and governments. Lately, implant manufacturers have also been mandated to perform postmarket surveillance with some hoping to base that on registry data. Aiming to assess the current status of knee-related registries, we performed a comprehensive literature and web search, which yielded 23 arthroplasty, 8 ligament, 4 osteotomy and 3 articular cartilage registries. Registries were evaluated for their scope, measured variables, impact and limitations. Registries have many advantages as they aim to increase awareness of outcomes; identify trends in practice over time, early failing implants, outlier surgeon or institution performance; and assist postmarketing surveillance. International collaborations have highlighted variations in practice. The limitations of registries are discussed in detail. Inconsistencies are found in collected data and measured variables. Potential measurement and selection biases are outlined. Without mandated data collection and with apparent issues such as unverified patient reporting of complications, registries are not designed to replace adverse event recording in place of a proper safety and efficacy study, as demanded by regulators. Registry 'big data' can provide evidence of associations of problems. However, registries cannot provide evidence of causation. Hence, without careful consideration of the data and its limitations, registry data are at risk of incorrectly drawn conclusions and the potential of misuse of the results. That must be guarded against. Looking at the future, registry operators benefit from a collective experience of running registries as they mature, allowing for improvements across specialties. Large-scale registries are not only of merit, improving with stakeholder acceptance, but also are critical in furthering our understanding of our patients' outcomes. In doing so, they are a critical element for our future scientific discourse.


A new integrative approach to assess aortic stenosis burden and predict objective functional improvement after TAVR.
A non-negligible rate of patients undergoing transcatheter aortic valve replacement (TAVR) do not report symptomatic improvement or even die in the short-midterm. We sought to assess the degree of objective functional recovery after TAVR and its prognostic implications and to develop a predictive model. In a cohort of patients undergoing TAVR, a prospective evaluation of clinical, anatomical, and physiological parameters was conducted before and after the procedure. These parameters were derived from echocardiography, non-invasive analysis of arterial pulse waves, and cardiac tomography. Objective functional improvement 6 months after TAVR was assessed using a 6-min walk test and nitro-terminal pro-brain natriuretic peptide (NT-proBNP) levels. The derived predictive model was prospectively validated in a different cohort. A clinical follow-up was conducted at 2 years. Among the 212 patients included, objective functional improvement was observed in 169 patients (80%) and subjective improvement in 187 (88%). Patients with objective functional improvement showed a much lower death rate at 2 years (9% vs. 31% p = 0.0002). Independent predictors of improvement were as follows: mean aortic gradient of ≥40 mmHg, augmentation index75 of ≥45%, the posterior wall thickness of ≤12 mm, and absence of atrial fibrillation. A simple integer-based point score was developed (GAPA score), which showed an area under the curve of 0.81 for the overall cohort and 0.78 for the low-gradient subgroup. In a validation cohort of 216 patients, these values were 0.75 and 0.76, respectively. A total of 80% of patients experienced objective functional improvement after TAVR, showing a significantly lower 2-year mortality rate. A predictive score was built that showed a good discriminative performance in overall and low-gradient populations.


[Venous thromboembolism in Cameroon (report of 18 cases)].
The purpose of this report was to describe clinical and ultrasound findings associated with venous thromboembolism of the lower extremities and pulmonary embolism observed in our department with special emphasis on the utility of venous ultrasound in the diagnosis of venous thromboembolism. Between January 1, 1998 and December 31, 1999, a total of 47 venous ultrasound procedures were carried out according to the standard technique using a Logic 400 MD system (General Electric). Deep venous thrombosis was diagnosed in 18 cases (38.3%). There were 10 men and 8 women with a mean age of 46 years (range, 24 to 71 years). Thrombosis involved the internal saphenous, popliteal, or sural vein in 12 cases, the common or deep femoral vein in 4, and the external iliac vein in 2. The most common risk factors observed in our series were surgery, predisposing conditions, history of venous thrombosis and morbid obesity (66.8% of case). Prolonged periods of bed confinement and neoplasm (lower extremity Kaposi's disease) were more uncommon (22.2%). Pulmonary embolism occurred during the observation period in six cases (33.3%) including 3 (50%) that were fatal. Based on these findings it can be concluded that although its incidence is relatively low in black African patients at risk, thromboembolic disease is often fatal and requires routine preventive treatment using heparin.


Adipocyte differentiation of human bone marrow-derived stromal cells is modulated by microRNA-155, microRNA-221, and microRNA-222.
Human mesenchymal stromal cells (hMSCs) are capable of limited self-renewal and multilineage differentiation in vitro. Several studies have demonstrated that microRNAs (miRNAs, miRs), post-transcriptional modifiers of mRNA stability and protein translation, play crucial roles in the regulation of these complex processes. To gain knowledge regarding the role of miRNAs in human adipocyte differentiation, we examined the miRNA expression profile of the immortalized human bone marrow-derived stromal cell line hMSC-Tert20. Such a model system has the advantage of a reproducible and consistent phenotype while maintaining important properties of the primary donor cells, including the potential to differentiate to adipocytes, osteoblasts, and chondrocytes. We identified 12 miRNAs that were differentially expressed during adipogenesis, of which several have been previously shown to play important roles in adipocyte biology. Among these, the expression of miRNA-155, miRNA-221, and miRNA-222 decreased during the adipogenic program of both immortalized and primary hMSCs, suggesting that they act as negative regulators of differentiation. Interestingly, ectopic expression of the miRNAs significantly inhibited adipogenesis and repressed induction of the master regulators PPARγ and CCAAT/enhancer-binding protein alpha. Our study provides the first experimental evidence that miRNA-155, miRNA-221, and miRNA-222 have an important function in human adipocyte differentiation, and that their downregulation is necessary to relieve the repression of genes crucial for this process.


Chronic bronchitis--alterations of the bronchial mucosa.
The mucociliary transport system is usually an important defense system which protects the body against a variety of noxious agents. Reactions of the bronchial mucosa to chronic infections are seen in the ciliated cells, the amount of globlet cells, in modifications of the basement membrane underlying the bronchial epithelium and an altered percentage of inflammatory cells. In ciliated cells the following atypia can be seen: thickening of the ciliar membrane, swollen cilia, formation of compound cilia, disarrangement of microtubules. Common alterations of the basement membrane are: increased diameter of the basement membrane zone, inhomogeneous staining pattern of the basement membrane zone, formation of cytoplasmatic protrusions, formation of double layers of the basement membrane and increased number of cytoplasmatic bound vesicles. Structural abnormalities of the basement membrane will lead to disturbances of the zone of transition and have to be interpreted as a sign of disregulation in the process of diffusion and resorption. The inflammatory response of the epithelium during chronic bronchitis and asthma shows many similarities. The bronchial epithelium has a specific reaction pattern which supports the response against different noxious agents. So all findings have to be interpreted as unspecific pathological changes. All alterations may show different degrees of severity and are dependent on individual pattern and the severity of chronic process. Electronmicroscopical examinations in combination with lightmicroscopical findings and immuno-histochemistry and seen in context with clinical data help to understand the mechanism of the inflammatory process.


Canine laparoscopic ovariectomy using two 3- and 5-mm portal sites: A prospective randomized clinical trial.
Laparoscopic ovariectomy (LapOve) was performed in 3 groups (2.7-mm/5-mm, 5-mm/3-mm, and 5-mm telescope/5-mm bipolar forceps) of small dogs (n = 60). Surgical times, bleeding rates, complications, and laparoscopic visualization were recorded and compared among groups. Use of the 3-mm bipolar forceps significantly increased the surgical time and showed higher bleeding rates compared with the 5-mm bipolar forceps. The 2.7-mm telescope significantly decreased the laparoscopic view. No complications were seen in any group. In conclusion, the 2.7-mm 30° telescope or the 3-mm bipolar forceps combined with the 5-mm instruments could be used as an alternative technique for LapOve in dogs up to 10 kg. The 2.7-mm telescope with the 5-mm bipolar forceps was the most efficient combination based on surgical time.


The Role of School in the Upward Mobility of Disadvantaged Immigrants' Children.
How can we explain exceptional advancement by disadvantaged immigrants' children? Extending segmented assimilation theory, this article traces the structural and relational attributes of high schools attended by young adults who reached their late twenties in 2000. Hypotheses are derived from theories in sociology of education and tested with four waves of data from the National Educational Longitudinal Study (NELS). The authors offer three major findings. First, an overwhelming majority of disadvantaged students attend public schools; some relational attributes are typical in public schools attended by disadvantaged students. Second, children's upward mobility is shaped by the structural and relational attributes of their high schools. Most school effects are the same for disadvantaged and advantaged youngsters, and student-educator bonds and curriculum structure have even stronger positive effects for the disadvantaged. Finally, mobility patterns differ widely among Chinese, Mexicans, and whites. Mexicans are less likely to be exposed to favorable school attributes.


Candida in pancreatic infection: a clinical experience.
Pancreatic infection remains a significant clinical problem, with substantial morbidity and mortality. Published case reports of Candida species identified in these infections prompted a review of 17 consecutive patients recently treated for peripancreatic infection by scheduled relaparotomy. Six patients were transferred from other hospitals, all having undergone prior operative intervention (median stay elsewhere: 58 days). The 11 other patients underwent initial operation an average of 14 days after admission. Candida species were identified in the initial operative cultures of 5 patients (29%), three of whom had undergone previous drainage at other hospitals. Two patients (11.7%) had Candida identified at subsequent operation. Six patients were treated with Amphotericin B for a median of 12 days (range 6-32) and a median dosage of 420 mg (range 225-830 mg). All patients were cleared of their Candida infection, but three subsequently died, for an overall mortality of 17.6%. Candida infected patients suffered a 42 per cent mortality. Our series supports the suspicion that Candida is much more frequent (41% of patients) than previously recognized in peripancreatic sepsis, and is commonly acquired after the initial operation. Amphotericin B therapy is effective in clearing Candida infection, but affected patients have a high associated mortality.


Cytomegalovirus Infection among Pregnant Women in Beijing: Seroepidemiological Survey and Intrauterine Transmissions.
Primary cytomegalovirus (CMV) infection during pregnancy can cause congenital defects. Available data for CMV infection during pregnancy in north China are inadequate. The aim of this study was to evaluate the epidemiology of maternal CMV infection and explore the incidence of congenital infection. In this prospective study, serum CMV IgG and IgM antibodies were measured in 2,887 pregnant women using ELISA, and the IgG avidity test was performed on all IgM-positive subjects. The seroprevalence of anti-CMV IgG was 94.70%, and of anti-CMV IgM was 1.28%. CMV IgG prevalence increased significantly with age (p < 0.01). Women living in downtown areas showed higher IgG prevalence than those residing in urban areas (p = 0.023). CMV-IgM seroprevalence was highest in autumn (p = 0.021). There was no difference in IgM seroprevalence by age, socioeconomic status, geographical area, or gravida. The rate of primary CMV infection was 0.45% (13/2,887) at the first trimester. The seroconversion rate during pregnancy was 0.76% (22/2,887). One woman underwent seroconversion during pregnancy and gave birth to an infant with asymptomatic CMV infection. Congenital CMV infection was diagnosed in five of the 14 infants from 14 mothers with active infection, for a vertical transmission rate of 35.71% (5/14). Three infants were asymptomatic, whereas two infants presented symptomatic infection with hearing deficits. Although CMV IgG prevalence is relatively high in north China, significant attention to primary CMV infection during pregnancy is still needed.


Dehydroepiandrosterone sulphate: diabolical hormone or epiphenomenon in aneurysmal subarachnoid hemorrhage?
Inflammation is purported to play an important role in the clinical course of subarachnoid hemorrhage. The current study by Höllig et al. entails using dehydroepiandrosterone sulfate, a hormone that inhibits key inflammatory pathways, as a predictor of functional outcome in these patients.


Cholic acid for hepatic steatosis in patients with lipodystrophy: a randomized, controlled trial.
Hepatic steatosis is a common complication in patients with lipodystrophies and can lead to cirrhosis. There is no proven effective therapy for hepatic steatosis, but cholic acid (CA), a farnesoid X receptor agonist, has previously been shown to reduce hepatic triglyceride (TG) content in mice and serum TG in humans. Our objective was to assess clinical efficacy and tolerability of CA therapy in patients with lipodystrophy and hepatic steatosis. A randomized, double-blind, placebo-controlled, crossover study. Eighteen patients with genetic or autoimmune lipodystrophies and elevated hepatic TG content participated in the study. The intervention was CA (15 mg/kg per day) compared with placebo for a period of 6 months each. Hepatic TG content, the primary outcome variable, was measured with (1)H magnetic resonance spectroscopy at baseline and at 3 and 6 months during each study period. Levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), and TG were secondary end points of the study. Compared with placebo, CA did not reduce (median (interquartile range) hepatic TG content (14.8% (9.4-19.0%) vs 15.9% (10.5-26.5%) respectively; P=0.42) or serum TG ((340 mg/dl (233-433 mg/dl) vs 390 mg/dl (233-595 mg/dl) respectively; P=0.45)). CA therapy also did not change AST, ALT, or GGT levels. Two patients developed diarrhea and excessive flatus while taking CA and these symptoms resolved after reducing the dose of CA. CA was well tolerated but did not reduce hepatic TG content in patients with lipodystrophy.


The role of intervention mapping in designing disease prevention interventions: A systematic review of the literature.
To assess the role of Intervention Mapping (IM) in designing disease prevention interventions worldwide. Systematic search and review of the relevant literature-peer-reviewed and grey-was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) guidelines. Only five of the twenty two included studies reviewed were RCTs that compared intervention using IM protocol with placebo intervention, and provided the outcomes in terms of percentage increase in the uptake of disease-prevention programmes, and only one of the five studies provided an effect measure in the form of relative risk (RR = 1.59, 95% CI = 1.08-2.34, p = 0.02). Of the five RCTs, three were rated as strong evidences, one as a medium evidence and one as a weak evidence, and they all reported statistically significant difference between the two study groups, with disease prevention interventions that have used the intervention mapping approach generally reported significant increases in the uptake of disease-prevention interventions, ranging from 9% to 28.5% (0.0001 ≤ p ≤ 0.02), On the other hand, all the 22 studies have successfully identified the determinants of the uptake of disease prevention interventions that is essential to the success of disease prevention programmes. Intervention Mapping has been successfully used to plan, implement and evaluate interventions that showed significant increase in uptake of disease prevention programmes. This study has provided a good understanding of the role of intervention mapping in designing disease prevention interventions, and a good foundation upon which subsequent reviews can be guided.


Interaction between Cardiac Functional Indices during Incremental Exercise Test Reveals the Peculiarities of Adaptation to Exercising.
Background and objectives: Physical load causes structural changes in the heart that vary depending on the type of training and may affect the function of the heart. Aim of the study: To determine, using the applied co-integration method on algebraic data, the impact of sprinting and of endurance adaptation on the dynamic interactions of cardiovascular functional indices while participants were performing under an increasing workload, up to their inability to continue. Materials and Methods: Healthy athletes were chosen to take part in this study and were separated into two groups: Sprinters (n = 11) and endurance athletes (n = 13). The bicycle ergometric method of incremental increase in a provocative workload (graded stress) was used. The heart rate, stroke volume, and cardiac output were determined using the tetrapolar rheography method. Results: Individuals who are adapted to endurance while carrying physical loads, in contrast to well adapted sprinters, are characterized with a lower rate of changing the pace of interactions between stroke volume and cardiac output while performing at an increasing workload up to their inability to continue. Also, endurance athletes displayed a long and relatively stable phase as well as a greater decrease of interaction between indices at the end of the workload. At the beginning of the exercise, the interaction between the stroke volume and the cardiac output was reduced. However, as the physical load continued, this interaction became significantly stronger. The comparison of the stroke volume and the cardiac output's dynamic interaction revealed that the endurance group had a greater working capacity. Conclusions: Typical dynamics of interactions during the testing with an increasing physical load can be differentiated into separate phases: The decrease of interaction at the onset of the load, the increase during the continuation of an increased workload, and the decrease at the last stages of the load.


Efficacy and Safety of Fingolimod in Daily Practice: Experience of an Academic MS French Center.
Fingolimod (Fg), a sphingosine 1-phosphate receptor modulator, decreases the annual relapse rate (ARR) in relapsing-remitting multiple sclerosis (RRMS). The aim of this study was to assess the efficacy and safety of Fg in daily practice in patients with RRMS, previously treated with natalizumab (Nz) or not, and systematically followed during at least 1 year. Data were collected from the patient files. Primary endpoint was the comparison between the ARR the year before Fg onset and after 1 and 2 years of Fg treatment. The secondary endpoints were the difference between Expanded Disability Status Scale (EDSS) at Fg onset and after 1 and 2 years of treatment, and safety. In the whole sample, we confirmed Fg efficacy on the ARR (0.895 before vs. 0.364 1 year after, p < 0.0001). Between our two groups (with or without Nz before Fg), the ARR was higher in the Nz group during the first year but similar during the second year. The EDSS was stable during the first year of Fg but significantly higher after 2 years (3.33 vs. 3.72, p = 0.02). Concerning safety, only three patients had to discontinue Fg because of tolerance issues. Our study showed that Fg is safe in RRMS and can be used either after first-line treatments or after Nz. However we observed a mild disability progression after 2 years.


[Roles of aberrant endothelial progenitor cells in pathogenesis of systemic sclerosis].
Systemic sclerosis (SSc) is a multisystem connective tissue disease characterized by excessive fibrosis and microvasculopathy, along with poor vascular formation and repair. The maintenance of the postnatal vascular system requires constant remodeling through vasculogenesis, which is mediated by the de novo differentiation of mature endothelial cells from endothelial progenitor cells (EPCs). However, a great deal of controversy about EPCs and their roles in postnatal vascular formation has arisen because of discrepancies in how EPCs are defined. The current consensus is that EPCs are heterogeneous cell population containing an extremely small count of "true EPCs", and pro-angiogenic hematopoietic cells (PHCs) that promotes vascular formation and repair through secretion of pro-angiogenic factors, and differentiation into endothelial cells and mural cells. In 2004, we reported a reduced number and impaired function of circulating CD34(+)CD133(+)CD309(+)CD45(dim)CD14(-) EPCs, which are now regarded as an immature subset of PHCs, in patients with SSc, and proposed a theory that defective vascular repair machinery as one of important mechanisms contributing to SSc vasculopathy. In addition, we showed that in SSc patients, circulating monocytic PHCs were increased and have enhanced angiogenic potency and differentiation potential to fibroblast-like cells. In summary, EPCs are involved in the pathogenesis of SSc by participating in two major pathological features, microvasculopathy and excessive fibrosis. Understanding the roles of EPCs in disease process of SSc may be key to dissecting its pathogenesis and to developing novel therapeutic strategies for this intractable condition.


PAF, rather than histamine, participates in mouse anaphylactic hypotension.
We determined the roles of platelet-activating factor (PAF) and histamine in anaphylactic hypotension in ovalbumin-sensitized anesthetized BALB/c mice. The effects of PAF and histamine on hemodynamic variables were studied by measuring the systemic arterial (Psa), portal venous (Ppv) and central venous (Pcv) pressures. Intravenous PAF evoked a biphasic Psa response, an initial rapid and transient drop followed by marked hypotension, accompanied by a decrease in Pcv. Histamine caused only mild systemic hypotension. Both agents similarly increased Ppv by approximately 4 cm H(2)O at high doses. After an injection of antigen, Psa initially increased slightly and then decreased from the baseline of 94 +/- 1 mm Hg to 46 +/- 1 mm Hg at 10 min after antigen administration, with Pcv decreasing by 2.5 cm H(2)O. Ppv increased by 3.5 cm H(2)O at 5 min after antigen injection. Pretreatment with either CV-6209 (PAF receptor antagonist, 1 mg/kg) or diphenhydramine (histamine H(1) receptor antagonist, 20 mg/kg) significantly attenuated an antigen-induced decrease in Psa. The inhibitory action of CV-6209 was greater than that of diphenhydramine, and the combination of these 2 antagonists almost completely inhibited the anaphylactic hypotension. In contrast, the antigen-induced increase in Ppv was attenuated by CV-6209 alone but augmented by diphenhydramine. It is concluded that anaphylactic hypotension is mainly mediated by PAF and, to a lesser extent, by histamine in anesthetized BALB/c mice.


Dissociation of vasoconstrictor and platelet aggregatory activities of thromboxane by carbocyclic thromboxane A2, a stable analog of thromboxane A2.
Carbocyclic thromboxane A2 [2 beta (Z),3 alpha- (1E,3R*)-3-(3-hydroxy(1-octenyl)-bicyclo[3.1.1]hept-2-yl-5-heptenoic acid], a stable analog of thromboxane A2, has been tested for its physiologic properties. Carbocyclic thromboxane A2 is a potent coronary vasoconstrictor, stimulating cornonary vascular smooth muscle at concentrations as low as 29 pM. At 1-5 micro M it is also an inhibitor of arachidonic-acid- and endoperoxide-induced aggregation of platelets. At 200 nM it stimulated the release of lysosomal hydrolases from large granule fractions of liver homogenate. It inhibited thromboxane synthesis in platelets, although it did not inhibit synthesis of prostacyclin in ram seminal vesicles. Thus, carbocyclic thromboxane A2, a molecule closely related to thromboxane A2, separates coronary vasoconstrictor from platelet-aggregating activity. The constrictor activity predominates in vivo; carbocyclic thromboxane A2 induces coronary vasoconstriction leading to myocardial ischemia and sudden death in rabbits in the absence of pulmonary or coronary thrombosis.


Silver nanodesert rose as a substrate for surface-enhanced Raman spectroscopy.
Silver galvanic displacement on silicon has been employed to produce large-area reproducible substrates, with morphology similar to that of the natural desert rose but on the micrometer scale. The process is based on an extremely simple wet chemistry approach using only AgF and KF, as silver and fluoride sources. A key element is the absence of HF in the deposition solution, which has been commonly used in previous silver galvanic displacement processes. The new process affords a higher degree of control in the redox reaction than those reported previously. The structures formed in this manner possess a large area-to-volume ratio with a high density of rough silver flakes uniformly distributed across the substrate. The silver morphology on the nanometer scale is shown to provide an excellent platform for surface-enhanced Raman spectroscopy (SERS), yielding detection levels for trans-1,2-bis(4-pyridyl)ethylene, 4-mercaptopyridine, and Rhodamine 6G in solution down to ppb, ppt, and ppq limits, respectively. The SERS reproducibility on the substrate was verified by monitoring the signal intensity variations across the sample. The simplicity of the substrate fabrication process, as well as the excellent uniformity, opens up opportunities for the quantitative and in-field chemical trace analysis using these substrates.


A hybrid method for identification of structural domains.
Structural domains in proteins are the basic units to form various proteins. In the protein's evolution and functioning, domains play important roles. But the definition of domain is not yet precisely given, and the update cycle of structural domain databases is long. The automatic algorithms identify domains slowly, while protein entities with great structural complexity are on the rise. Here, we present a method which recognizes the compact and modular segments of polypeptide chains to identify structural domains, and contrast some data sets to illuminate their effect. The method combines support vector machine (SVM) with K-means algorithm. It is faster and more stable than most current algorithms and performs better. It also indicates that when proteins are presented as some Alpha-carbon atoms in 3D space, it is feasible to identify structural domains by the spatially structural properties. We have developed a web-server, which would be helpful in identification of structural domains (http://vis.sculab.org/~huayongpan/cgi-bin/domainAssignment.cgi).


Enhanced corticosterone-binding capacity in plasma after stimulation of the rat adrenal cortex: the possibility of a simultaneous release of protein and corticosterone.
Exposure of rats to either footshock or handling stress produced a significant increase in both plasma corticosterone concentration and specific binding capacity. Non-specific binding was eliminated using the synthetic glucocorticoid, dexamethasone. The increase in both plasma corticosterone and specific binding capacity was biphasic following exposure to footshock. Adrenalectomy and pretreatment with betamethasone abolished both phases of the enhanced binding capacity and plasma steroid concentration. Intraperitoneal injection of ACTH (1-24) in animals pretreated with betamethasone resulted in a biphasic rise in plasma concentrations of corticosterone but only the initial increase in binding capacity. Dissociation constant (Kd) values, determined by Scatchard analysis, for adrenalectomized and betamethasone-pretreated animals were 546 and 556 pmol/l respectively. These values were significantly different from the Kd in animals with functional adrenals (631 pmol/l). The results are discussed in the light of a possible specific corticosteroid-binding globulin (CBG) like binding protein of adrenal origin released in conjunction with corticosterone. This binding protein has a lower affinity for corticosterone and a shorter half-life than CBG.


Serum Fibrinogen as a Prognostic Factor in Sudden Sensorineural Hearing Loss: A Meta-analysis.
High blood viscosity has been proposed as a mechanism for sudden sensorineural hearing loss (SSNHL); however, the relationship between blood markers of fibrinolysis or coagulation and severity or prognosis of SSNHL is still unclear. The aim of this study is to investigate the relationship between serum fibrinogen and SSNHL. PubMed and Scopus were searched for English language articles using the following keywords: SSNHL, sudden hearing loss, sudden deafness, idiopathic hearing loss or idiopathic sensorineural hearing loss, and fibrinogen. The articles in the study related to SSNHL and provided data about the serum fibrinogen level. The data included patient profiles, fibrinogen level, recovery, and treatment modality. Nineteen articles were selected. The aggregated data were analyzed using the random effect model. Two articles that included the fibrinogen level with recovery rates were analyzed for the relationship between the fibrinogen level and recovery. The average fibrinogen level was 318 ± 8.5 mg/dl (mean ± standard error, within normal range). There was no difference in the fibrinogen level between SSNHL patients and the control group. The fibrinogen level of the recovery group was lower than that of the no recovery group. This showed that a high fibrinogen level was associated with poor prognosis, and it could be related to the severity of the pathological change rather than being the cause of the SSNHL. SSNHL includes various pathologies; therefore, the appropriate therapy should be selected based on each condition.


Nutritional status as a predictor of child survival: summarizing the association and quantifying its global impact.
By pooling the results from five previously published prospective studies, we have obtained estimates of the relative risks of mortality among young children 6-24 months after they had been identified as having mild-to-moderate or severe malnutrition. These risk estimates, along with global malnutrition prevalence data, were then used to calculate the total number of young-childhood deaths "attributable" to malnutrition in developing countries. Young children (6-60 months of age) with mild-to-moderate malnutrition (60-80% of the median weight-for-age of the reference population) had 2.2 times the risk of dying during the follow-up period than their better nourished counterparts (> 80% of the median reference weight-for-age). Severely malnourished young children (< 60% of the reference median weight-for-age) had 6.8 times the risk of dying during the follow-up period than better nourished children. Each year approximately 2.3 million deaths of young children in developing countries (41% of the total for this age group) are associated with malnutrition. The comparability of studies, methods used to derive pooled values, potentially confounding factors that may influence risk estimates, and the validity of the results are discussed. Child survival programmes should assign greater priority to the control of childhood malnutrition.


Reconstructive surgery in the myelomeningocele hip.
In 17 myelomeningocele patients with subluxating or dislocated hips, 31 posterolateral iliopsoas were treated by transfers, as described by Sharrard, in addition to complimentary procedures to balance muscle power and center the hip joint. Surgical goals are to stablize the hips within the acetabulum and prevent the occurrence of severe fixed flexion and adduction contractures of the hip and secondary lumbar lordosis. Surgery is performed at about 1 year of age with bilateral adductor transfer to the ischium, followed in 2 weeks by the posterolateral transfer of the iliopsoas muscle, first on one hip and 2 weeks later following with the other hip. Adductor transfer to the ischium reduces the adductor and secondary flexor power of the adductor longus and anterior fibers of the adductor brevis. After transfer it provides some extensor power. The range of abduction is increased. The transferred iliopsoas muscle has a better mechanical advantage in its new function as an abductor and extensor, and the hips were better centered radiographically than the cases with iliopsoas transfer alone. Fewer bony procedures were required to center the hips when the iliopsoas transfer was performed in children before the age of 1 1/2 years.


Percutaneous closure of an aortopulmonary window in a young adult patient: a case report of transcatheter closure with an occluder device.
Aortopulmonary window is a congenital heart disease requiring a surgical approach to achieve a complete correction. We report on a case of a young female symptomatic adult patient with a significant aortopulmonary window. The patients refused surgical approach and since the defect was far enough from the pulmonary and aortic valve a percutaneous approach was proposed. The large aortopulmonary window was closed effectively with an Amplatzer Muscular Septal Occluder device (Abbott, Plymouth, MN, USA). At 1-year follow-up, the device was in a good position with no residual shunt. Transcatheter closure of an aortopulmonary window is an effective and safe procedure in specific anatomic defects.


Pleiotropic hubs drive bacterial surface competition through parallel changes in colony composition and expansion.
Bacteria commonly adhere to surfaces where they compete for both space and resources. Despite the importance of surface growth, it remains largely elusive how bacteria evolve on surfaces. We previously performed an evolution experiment where we evolved distinct Bacilli populations under a selective regime that favored colony spreading. In just a few weeks, colonies of Bacillus subtilis showed strongly advanced expansion rates, increasing their radius 2.5-fold relative to that of the ancestor. Here, we investigate what drives their rapid evolution by performing a uniquely detailed analysis of the evolutionary changes in colony development. We find mutations in diverse global regulators, RicT, RNAse Y, and LexA, with strikingly similar pleiotropic effects: They lower the rate of sporulation and simultaneously facilitate colony expansion by either reducing extracellular polysaccharide production or by promoting filamentous growth. Combining both high-throughput flow cytometry and gene expression profiling, we show that regulatory mutations lead to highly reproducible and parallel changes in global gene expression, affecting approximately 45% of all genes. This parallelism results from the coordinated manner by which regulators change activity both during colony development-in the transition from vegetative growth to dormancy-and over evolutionary time. This coordinated activity can however also break down, leading to evolutionary divergence. Altogether, we show how global regulators function as major pleiotropic hubs that drive rapid surface adaptation by mediating parallel changes in both colony composition and expansion, thereby massively reshaping gene expression.


Phaseless three-dimensional optical nanoimaging.
We propose a method for optical nanoimaging in which the structure of a three-dimensional inhomogeneous medium may be recovered from far-field power measurements. Neither phase control of the illuminating field nor phase measurements of the scattered field are necessary. The method is based on the solution to the inverse scattering problem for a system consisting of a weakly-scattering dielectric sample and a strongly-scattering nanoparticle tip. Numerical simulations are used to illustrate the results.


High prevalence of metabolic syndrome in an elderly Croatian population - a multicentre study.
To investigate the prevalence and characteristics of metabolic syndrome (MetS) in a healthy elderly Croatian population. Cross-sectional study consisting of a health check including anthropometric measures and food questionnaires as well as analysis of biochemical parameters related to MetS. The diagnostic criteria of the International Diabetes Federation (IDF) were used for diagnosis of MetS. Four centres in continental (Virovitica and Zagreb) and Adriatic coast (Split and Omiš) regions of Croatia. Free-living elderly persons aged 70-90 years (n 320). Significantly lower MetS prevalence was found among participants from small urban centres compared with those from large urban centres (59·1 % v. 69·6 %; P = 0·051). Participants without MetS consumed wine more frequently (P = 0·05) than those with MetS. Compared with their peers with HDL cholesterol (HDL-C) <1·03 mmol/l, more male participants with HDL-C ≥1·03 mmol/l consumed wine (P = 0·04) or pelagic fish (P = 0·03). The prevalence of participants with TAG ≥1·7 mmol/l was higher in wine non-consumers (P = 0·05) than in wine consumers. Multivariate analysis with age and gender as covariates showed a significant inverse association of wine consumption with total cholesterol (P < 0·001), a positive association with HDL-C (P < 0·001) and a marginally inverse association with TAG (P = 0·06). In the male population, alkaline phosphatase and γ-glutamyl transferase activities were higher in participants with MetS (P < 0·05). High MetS prevalence was observed in an elderly Croatian population. Data showed that moderate consumption of wine and/or pelagic fish has a protective role against MetS in the population studied.


Sequence analysis of cymbidium ringspot virus satellite and defective interfering RNAs.
The nucleotide sequences of cloned cDNA copies of satellite and defective interfering (DI) RNAs of cymbidium ringspot virus were determined. DI RNA is 499 nucleotides long and is composed of six stretches of sequence derived from CyRSV genomic RNA. Four of these stretches share common 5' sequences and the 5' and 3' ends are identical to those of genome RNA. Satellite RNA is 621 nucleotides long. In some regions the sequence is very similar (60% to 100% identical) to genomic RNA. A consensus sequence is proposed to be involved in the replication of genomic, DI and satellite RNAs.


Comparison of maximal oxygen consumption between obese black and white adolescents.
The purpose of this investigation was to determine whether maximal oxygen consumption (VO2max) differed between clinically obese black and white children and if a difference existed to determine whether it was related to hematological profiles and/or physical activity/inactivity levels. Twenty-three black and 21 white adolescents were matched for age, BMI, and Tanner stage (II-V). Body composition was determined by DEXA and CT scan. Daily physical activity/inactivity was assessed by questionnaire. VO2max was assessed using the Bruce treadmill protocol. Black participants had significantly lower VO2max and VO2maxFFM values when compared with white adolescents (26.1 +/- 4.2 versus 29.9 +/- 3.1 mL . kg(-1) . min(-1); 48.3 +/- 8.8 versus 55.6 +/- 5.2 mL . kgFFM(-1) . min(-1), respectively). Black adolescents also had significantly lower Hb concentrations ([Hb]) than white children (12.7 +/- 1.3 versus 13.4 +/- 0.7 g/dL). Black adolescents were more physically inactive than their white peers. VO2max correlated with [Hb] for the combined groups. Obese black adolescents had lower VO2max compared with white children and this difference was explained, in part, by the lower [Hb] observed in the black participants. Further investigations should study Hb flow rate (a function of [Hb] . maximal cardiac output) and physical activity/inactivity patterns in obese black and white children as it relates to VO2max.


Dual modification of biomolecules.
With the advent of novel bioorthogonal reactions and "click" chemistry, an increasing number of strategies for the single labelling of proteins and oligonucleotides have emerged. Whilst several methods exist for the site-selective introduction of a single chemical moiety, site-selective and bioorthogonal dual modification of biomolecules remains a challenge. The introduction of multiple modules enables a plethora of permutations and combinations and can generate a variety of bioconjuguates with many potential applications. From de novo approaches on oligomers to the post-translational functionalisation of proteins, this review will highlight the main strategies to dually modify biomolecules.


Public health. Grand Challenges in Global Health.
This week an international panel announces a list of 14 Grand Challenges in Global Health, and scientists throughout the world will be invited to submit grant proposals to pursue them with funds provided by the Bill and Melinda Gates Foundation. We describe the characteristics of these challenges and the process by which they were formulated and selected after receiving over 1000 responses to a "call for ideas" from the scientific community.


The Masquelet Technique: Can Disposable Polypropylene Syringes be an Alternative to Standard PMMA Spacers? A Rat Bone Defect Model.
Usually, the two-stage Masquelet induced-membrane technique for extremity reconstruction begins with a polymethylmethacrylate (PMMA) cement spacer-driven membrane, followed by an autologous cancellous bone graft implanted into the membrane cavity to promote healing of large bone defects. In exceptional cases, spacers made of polypropylene disposable syringes were successfully used instead of the usual PMMA spacers because of a PMMA cement shortage caused by a lack of resources. However, this approach lacks clinical evidence and requires experimental validation before being recommended as an alternative to the conventional technique. To (1) develop and (2) validate a critical-sized femoral defect model in rats for two stages of the Masquelet technique and to (3) compare the biological and bone healing properties of polypropylene-induced membranes and PMMA-induced membranes in this model. Fifty male Sprague Dawley rats aged 8 weeks old received a 6-mm femur defect, which was stabilized with an external fixator that was converted into an internal device. In the development phase, the defect was filled with PMMA in 16 rats to determine the most favorable timing for bone grafting. Two rats were excluded since they died of anesthetic complications. The other 14 were successively euthanized after 2 weeks (n = 3), 4 weeks (n = 4), 6 weeks (n = 4), and 8 weeks (n = 3) for induced membrane analyses. In the validation phase, 12 rats underwent both stages of the procedure using a PMMA spacer and were randomly assigned to two groups, whether the induced membrane was preserved or removed before grafting. To address our final objective, we implanted either polypropylene or PMMA spacers into the defect (Masquelet technique Stage 1; n = 11 rats per group) for the period established by the development phase. In each group, 6 of 11 rats were euthanized to compare the biological properties of polypropylene-induced membranes and PMMA-induced membranes using histological qualitative analysis, semiquantitative assessment of the bone morphogenic protein-2 content by immunostaining, and qualitative assessment of the mesenchymal stromal cell (MSC; CD31-, CD45-, CD90+, and CD73+ phenotypes) content by flow cytometry. Quantitative measurements from serum bone turnover markers were also performed. The five remaining rats of each group were used for Masquelet technique Stage 2, in which rat bone allografts were implanted in the induced membrane cavity after the polypropylene or PMMA spacers were removed. These rats recovered for 10 weeks before being euthanized for microCT quantitative measurements and bone histology qualitative assessment to evaluate and compare the extent of bone regeneration between groups. Induced membrane analyses together with serum bone turnover measurements indicated that a 4-week interval time between stages was the most favorable. Removal of the induced membrane before grafting led to almost constant early implant failures with poor bone formation. Four-week-old rats with polypropylene-triggered induced membranes displayed similar histologic organization as rats with PMMA-driven induced membranes, without any difference in the cell density of the extracellular matrix (4933 ± 916 cells per mm2 for polypropylene versus 4923 ± 1284 cells per mm2 for PMMA; p = 0.98). Induced membrane-derived MSCs were found in both groups with no difference (4 of 5 with polypropylene versus 3 of 3 with PMMA; p > 0.99). Induced membrane bone morphogenic protein-2 immunolabeling and serum bone turnover marker levels were comparable between the polypropylene and PMMA groups. MicroCT analysis found that bone regeneration in the polypropylene group seemed comparable with that in the PMMA group (29 ± 26 mm3 for polypropylene versus 24 ± 18 mm3 for PMMA; p > 0.99). Finally, qualitative histological assessment revealed a satisfactory endochondral ossification maturation in both groups. Using a critical-sized femoral defect model in rats, we demonstrated that polypropylene spacers could induce membrane encapsulation with histologic characteristics and bone regenerative capacities that seem like those of PMMA spacers. In a same bone site, polymers with close physical properties seem to lead to similar foreign body reactions and induce encapsulating membranes with comparable bone healing properties. Polypropylene spacers made from disposable syringes could be a valuable alternative to PMMA. These results support the possibility of a cementless Masquelet technique in cases of PMMA shortage caused by a lack of resources.


Mume Fructus water extract inhibits pro-inflammatory mediators in lipopolysaccharide-stimulated macrophages.
Mume Fructus (Family Rosaceae) is used as a traditional drug and health food in Asian countries. However, its therapeutic mechanisms and effects on macrophage-mediated inflammation remain unknown. In this study we examined the effect of Mume Fructus water extract (MFWE) on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The investigation focused on whether MFWE inhibited nitric oxide (NO) and prostaglandin (PG) E2 productions, as well as the expressions of inducible NO synthase (iNOS), cyclooxygenase (COX)-2, interleukin (IL)-6, nuclear factor-kappaB (NF-kappaB), and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW 264.7 cells. We found that MFWE inhibited LPS-induced NO, PGE(2), and IL-6 productions as well as the expressions of iNOS and COX-2. Furthermore, MFWE suppressed the LPS-induced phosphorylations of p38 MAPK and extracellular signal-regulated kinase MAPK, as well as IkappaBalpha degradation and NF-kappaB activation. These results suggest that MFWE has inhibitory effects on LPS-induced PGE2, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the phosphorylation of MAPKs following IkappaBalpha degradation and NF-kappaB activation.


RelB attenuates cigarette smoke extract-induced apoptosis in association with transcriptional regulation of the aryl hydrocarbon receptor.
Chronic obstructive pulmonary disease (COPD) is a chronic and prevalent respiratory disease caused primarily by long term inhalation of cigarette smoke. A major hallmark of COPD is elevated apoptosis of structural lung cells including fibroblasts. The NF-κB member RelB may suppress apoptosis in response to cigarette smoke, but its role in lung cell survival is not known. RelB may act as a pro-survival factor by controlling the expression of superoxide dismutase 2 (SOD2). SOD2 is also regulated by the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor that suppresses cigarette smoke-induced apoptosis. As the AhR is also a binding partner for RelB, we speculate that RelB suppresses cigarette smoke-induced apoptosis by regulating the AhR. Using an in vitro model of cigarette smoke exposure (cigarette smoke extract [CSE]), we found that CSE down-regulated RelB expression in mouse lung fibroblasts, which was associated with elevated levels of cleaved PARP. Genetic ablation of RelB elevated CSE-induced apoptosis, including chromatin condensation, and reduced mitochondrial function. There was also more reactive oxygen species production in RelB-/- cells exposed to CSE. While there was no alteration in Nrf2 expression or localization between RelB-/- and wild type cells in response to CSE, RelB-/- cells displayed significantly decreased AhR mRNA and protein expression, concomitant with loss of AhR target gene expression (Cyp1a1, Cyp1b1, Nqo1). Finally, we found that RelB binds to the Ahr gene at 3 sites to potentially increase its expression via transcriptional induction. These data support that RelB suppresses cigarette smoke-induced apoptosis, potentially by increasing the AhR. Together, these two proteins may comprise an important cell survival signaling pathway that reduces apoptosis upon cigarette smoke exposure.


Poly-l-Lactic Acid Nanofiber-Polyamidoamine Hydrogel Composites: Preparation, Properties, and Preliminary Evaluation as Scaffolds for Human Pluripotent Stem Cell Culturing.
Electrospun poly-l-lactic acid (PLLA) nanofiber mats carrying surface amine groups, previously introduced by nitrogen atmospheric pressure nonequilibrium plasma, are embedded into aqueous solutions of oligomeric acrylamide-end capped AGMA1, a biocompatible polyamidoamine with arg-gly-asp (RGD)-reminiscent repeating units. The resultant mixture is finally cured giving PLLA-AGMA1 hydrogel composites that absorb large amounts of water and, in the swollen state, are translucent, soft, and pliable, yet as strong as the parent PLLA mat. They do not split apart from each other when swollen in water and remain highly flexible and resistant, since the hydrogel portion is covalently grafted onto the PLLA nanofibers via the addition reaction of the surface amine groups to a part of the terminal acrylic double bonds of AGMA1 oligomers. Preliminary tested as scaffolds, the composites prove capable of maintaining short-term undifferentiated cultures of human pluripotent stem cells in feeder-free conditions.


Switching from single-stranded to double-stranded DNA limits the unwinding processivity of ring-shaped T7 DNA helicase.
Phage T7 helicase unwinds double-stranded DNA (dsDNA) by encircling one strand while excluding the complementary strand from its central channel. When T7 helicase translocates on single-stranded DNA (ssDNA), it has kilobase processivity; yet, it is unable to processively unwind linear dsDNA, even 60 base-pairs long. Particularly, the GC-rich dsDNAs are unwound with lower amplitudes under single-turnover conditions. Here, we provide evidence that T7 helicase switches from ssDNA to dsDNA during DNA unwinding. The switching propensity is higher when dsDNA is GC-rich or when the 3'-overhang of forked DNA is <15 bases. Once helicase encircles dsDNA, it travels along dsDNA and dissociates from the end of linear DNA without strand separation, which explains the low unwinding amplitude of these substrates. Trapping the displaced strand with ssDNA binding protein or changing its composition to morpholino oligomer that does not interact with helicase increases the unwinding amplitude. We conclude that the displaced strand must be continuously excluded and kept away from the central channel for processive DNA unwinding. The finding that T7 helicase can switch from ssDNA to dsDNA binding mode during unwinding provides new insights into ways of limiting DNA unwinding and triggering fork regression when stalled forks need to be restarted.


Clival chordoma.
Chordomas are uncommon tumors arising from embryonic remnats of the fetal notochord. The spheno-occipital and sacrococcygeal regions account for over 90 per cent of cases, the remaining 10 per cent occurring anywhere along the vertebral axis. A case of chordoma arising from the clivus is presented with a view to discussing management. The ability to obtain reasonable survival times associated with a useful and meaningful quality of life is emphasized. This may be accomplished by repeated, judicious surgical intervention with surprising ease and little morbidity to the patient in spite of its intracranial location.


[Treatment results of concurrent chemoradiotherapy with CDDP or CDGP plus 5-FU for carcinoma of the oropharynx and hypopharynx].
Twenty-eight patients with oropharyngeal or hypopharyngeal carcinoma received concurrent chemoradiotherapy with CDDP or CDGP plus 5-FU between January 2001 and April 2006. The numbers of patients according to clinical stage were stage II:2; stage III:5; stage IVa:19 ; and stage IVb:2. Total radiation dose was 60-73.8 Gy (median 66 Gy) and overall treatment time was 41-57 days (median 47 days). Two courses of 5-FU 700 mg/m(2) on days 1-5 and CDDP or CDGP 70 mg/m(2) on day 4 were administered concurrently with radiotherapy. Median follow-up period was 26 months (range, 8-64 months). The incidences of grade 3 or greater acute toxicity were leukopenia 29%, anemia 21%, thrombocytopenia 7%, pharyngeal mucositis 43% and nausea 14%. No severe late toxicity was observed. Treatment responses of primary lesions were CR in 24 patients (86%) and PR in 4 patients (14%). The two-year local control rate was 87% and the 2-year overall survival rate was 72%. Concurrent chemoradiotherapy with CDDP or CDGP plus 5-FU seemed to be effective for advanced carcinomas of the oropharynx and hypopharynx.


Changes in the weight of the adrenal glands and in the concentration of plasma corticosterone in perinatal rats after prenatal treatment with oestadiol benzoate.
On day 20 of gestation, foetal rats received a subcutaneous injection of 0-01 mg oestradiol benzoate dissolved in 0-05 ml sesame oil; foetuses in other litters were given sesame oil alone. Autopsy was performed on day 22 of gestation, at delivery or at various times after birth. Gravimetric and histological observations of the adrenals from oestradiol-treated, oil-treated and intact litter-mate control foetal and neonatal rats were performed together with determination of plasma corticosterone concentrations. Activity of 3 phi-ol dehydrogenase was also examined histochemically in the adrenals from these animals. The results indicated that oestradiol benzoate when given prenatally prevented the neonatal decline of adrenal weight and adrenocortical cell size. In normal or oil-treated rats, plasma corticosterone concentration was greatly increased during delivery and 2 h after birth, declining up to 12 h after birth. Oestradiol benzoate prevented this perinatal increase and suppressed the activity of 3 phi-ol dehydrogenase in the perinatal adrenal cortices.


New evidence on the effects of tea on mercury metabolism in humans.
The authors present the results of an experiment in which they explored the role of tea in human metabolic processing of methylmercury (MeHg) from fish consumption. The experiment involved 50 scientists from the Collaborative Mercury Research Network (COMERN) who agreed to eat fish for 2 daily meals for 3 consecutive days. Half of the participants also drank 6 cups of tea daily, starting a week before and continuing through the experiment. The authors calculated the total amount of MeHg that each participant ingested from (1) the measured mercury (Hg) level in fish and (2) the quantity of fish eaten, and compared it with the total increases of Hg and MeHg levels in participants' blood. Results indicated that the control group metabolized roughly 100% of the available fish MeHg, whereas the tea-exposed group showed blood levels of MeHg at more than 40% than that available in the fish provided, suggesting that an external MeHg pool supplied part of the measured blood MeHg increase. The authors conclude that tea may accelerate the enterohepatic MeHg cycle and contribute to a temporary bioamplification of MeHg in the bloodstream.


[Psychovegetative features in patients with biliary dyskinesia].
The aim was to study psychovegetative disorders in hypomotor and hypermotor forms of biliary diskinesia (BDK) in young patients. Psychological status and vegetative nervous function were studied in 62 patients with hypoBDK, 58 patients with hyperBDK and 33 control patients. According to AMPI, Spilberger's, Aizek's, Heck-Hess' tests, the psychological status of the patients from both groups was similar, while vegetative status in the hypoBDK group at rest and active orthostasis had a sympathetic trend. In hyperBDK group at rest sympathetic impacts enhanced, in stress these impacts intensified reaching higher values than in the control group. For the treatment of various forms of BDK, it is recommended to correct psychovegetative deviations with consideration of diskinesia forms.


Utility of Transient Elastography in Estimating Hepatic Iron Concentration in Comparison to Magnetic Resonance Imaging in Patients Who are Transfusion-Dependent: A Canadian Center Experience.
Transfusion-dependent hereditary anemias such as β-thalassemia (β-thal), predispose patients to iron overload and its numerous clinical sequelae. Accurate assessment of overall iron status and prompt initiation of chelation therapy to prevent irreversible end-organ damage can be achieved using magnetic resonance imaging (MRI) to measure liver iron concentration (LIC) as a surrogate marker of total body iron; however, its access may be associated with long wait times and delay in treatment. We report an observational cohort study at a single tertiary care center assessing the theoretical role of transient elastography (TE), which measures liver stiffness, in estimating LIC compared to other established diagnostic measures. While regression analyses confirm a moderate correlation between LIC per R2 MRI and serum ferritin level (pooled estimate of correlation = 0.55), there was no significant correlation between TE reading and LIC based on R2 MRI (pooled estimate of correlation = -0.06), and only a weak correlation was observed with serum ferritin level (pooled estimate of correlation = 0.45). These results suggest TE may not be sensitive enough to detect subtle changes in the hepatic parenchymal stiffness associated with liver iron deposition.


Evaluation of stress and pain in young children with cerebral palsy during early developmental intervention programs: a descriptive study.
The aim of this study was to use the Face, Legs, Activity, Cry, Consolability Scale; salivary cortisol levels; and withdrawal reflex thresholds to assess pain, stress, and pain sensitivity in young children with cerebral palsy during early developmental intervention programs. A total of 40 children with cerebral palsy (age range, 1-4 yrs) participated in the early intervention programs, which included neurodevelopmental treatment, neuromuscular electrical stimulation, occupational therapy, head acupuncture, and Chinese traditional manipulation five times per week for 3 wks. The Face, Legs, Activity, Cry, Consolability Scale was applied during the course of each treatment, and salivary cortisol samples were obtained from each child 10 mins before and 10 mins after each treatment. Withdrawal reflex thresholds were assessed via mechanical stimulation of the foot with von Frey hairs. All treatment programs caused some degree of pain. In descending order, the extents of the pain caused by each treatment were head acupuncture, neurodevelopmental treatment, neuromuscular electrical stimulation, Chinese traditional manipulation, and occupational therapy. There were statistically significant increases in salivary cortisol levels after the head acupuncture (P < 0.001), neurodevelopmental treatment (P < 0.001), neuromuscular electrical stimulation (P < 0.001), and Chinese traditional manipulation (P < 0.001) treatments. No significant changes were found in the withdrawal reflex thresholds during the study (P > 0.05). The results of this study demonstrate that early developmental intervention programs cause pain and stress in young children with cerebral palsy.


Mixed periapical lesion: an atypical radicular cyst with extensive calcifications.
The radicular cyst is an inflammatory odontogenic cyst of endodontic origin. Radiographically, the lesion appears as a periapical radiolucent image. This report describes a very rare case of a mixed periapical radiographic image diagnosed as a radicular cyst. A 37-year-old female patient presented a mixed, well-circumscribed image located in the periapical region of the left maxillary central incisor, which presented unsatisfactory endodontic treatment. Microscopic examination revealed a cavity lined by non-keratinized squamous epithelium and extensive calcifications in the cystic lumen and lining epithelium. Diagnosis of radicular cyst with extensive calcifications was established. Endodontic retreatment was performed and no radiographic signs of recurrence were observed 18 months after treatment. Although very rare, a radicular cyst should be considered in the differential diagnosis of a mixed periapical image associated to teeth with pulp necrosis.


Assessment of correlation between knee notch width index and the three-dimensional notch volume.
This study was done to determine whether there is a correlation between the notch volume and the notch width index (NWI) as measured on the three most frequently used radiographic views: the Holmblad 45 degrees, Holmblad 70 degrees, and Rosenberg view. The notch volume of 20 cadaveric knees was measured using Computed Tomography (CT). The Holmblad 45 degrees, Holmblad 70 degrees, and Rosenberg notch view radiographs were digitally re-created from the CT scans for each specimen, and the NWI was measured by two observers. The Pearson correlation coefficient between the NWI and notch volume was calculated, as well as between the three views. An independent t test was performed to determine the difference in NWI and notch volume between male and female specimens. The reliability for each view was also determined. There was no correlation between the NWI as measured on the Holmblad 45 degrees, Holmblad 70 degrees, or Rosenberg view and the notch volume. All three radiographic views proved reliable, but showed only a moderate correlation with each other. Men had larger notch volumes than women, but there was no difference in NWI. A knee with a small intercondylar notch is often considered an increased risk for ACL rupture. The NWI is a frequently used two-dimensional method to determine notch size. However, in the present study, this index was not positively correlated with the overall volume of the notch. Based on the results of the current study, the authors would advice to use caution when using notch view radiographs in a clinical setting to predict risk of ACL rupture.


The two-component model of memory development, and its potential implications for educational settings.
We recently introduced a two-component model of the mechanisms underlying age differences in memory functioning across the lifespan. According to this model, memory performance is based on associative and strategic components. The associative component is relatively mature by middle childhood, whereas the strategic component shows a maturational lag and continues to develop until young adulthood. Focusing on work from our own lab, we review studies from the domains of episodic and working memory informed by this model, and discuss their potential implications for educational settings. The episodic memory studies uncover the latent potential of the associative component in childhood by documenting children's ability to greatly improve their memory performance following mnemonic instruction and training. The studies on working memory also point to an immature strategic component in children whose operation is enhanced under supportive conditions. Educational settings may aim at fostering the interplay between associative and strategic components. We explore possible routes towards this goal by linking our findings to recent trends in research on instructional design.


A Quantitative Diagnostic Method for Phlegm and Blood Stasis Syndrome in Coronary Heart Disease Using Tongue, Face, and Pulse Indexes: An Exploratory Pilot Study.
Objectives: The aim of this study was to establish a quantitative syndrome differentiation model with logistic regression analysis for phlegm and blood stasis syndrome (PBSS) in coronary heart disease (CHD) to offer methodology guidance for the quantitative syndrome differentiation of Traditional Chinese Medicine (TCM). Design: Tongue, face, and pulse information of each subject was obtained using the TCM-intelligent diagnosis instruments. Logistic regression model was used to construct the syndrome diagnosis model. The area under receiver operating characteristic curve (ROC-AUC) was used to evaluate the diagnostic value of the model. Subjects: Among the 141 subjects, 83 belonged to the PBSS group, and 58 belonged to the non-PBSS group. Results: The independent indexes used to predict PBSS in patients with CHD were length of the crack (LC) (p = 0.002), number of ecchymosis (NE) (p < 0.001), length of philtrum (LEP) (p = 0.022), and right hand pulse h1 (Rh1) (p = 0.021). The expression of combining predictor L in this study was L = LC +57.58 NE +4.53 LEP +2.68 Rh1. The ROC curve analysis indicated that the AUC values of LC, NE, LEP, and Rh1 were 0.646, 0.710, 0.619, and 0.613, respectively. The AUC = 0.825 of the syndrome diagnosis model was the largest. Conclusions: The quantitative study of TCM syndrome based on logistic regression analysis provides a good method for the objective analysis and application of TCM syndrome.


Functionalization of Silk with In-Situ Synthesized Platinum Nanoparticles.
After platinum nanoparticles (PtNPs) were in-situ synthesized on silk fabrics through heat treatment, it was determined that the treatment of the silk fabrics with PtNPs imparted multiple functions, including coloring, catalysis, and antibacterial activity. The formation of PtNPs on fabrics was affected by the Pt ion concentration, pH value of solution, and reaction temperature. Acidic condition and high temperature were found to facilitate the formation of PtNPs on silk. The color strength of silk fabrics increased with the concentration of Pt ions. The PtNP treated silk fabrics exhibited reasonably good washing color fastness and excellent rubbing color fastness. The morphologies and chemical components of the treated silk fabrics were analyzed using scanning electron microscopy and X-ray photoelectron spectroscopy. The PtNP treated silk fabric exhibited significant catalytic function and a notable antibacterial effect against Escherichia coli (E. coli).


Consumption of folic acid-fortified bread improves folate status in women of reproductive age in Chile.
Since January 2000 the Chilean Ministry of Health has required the fortification of wheat flour with folic acid (FA) at a concentration of 2.2 mg FA/kg in order to reduce the risk of neural tube defects (NTD) in newborns. This policy was expected to result in a mean additional intake of approximately 400 microg FA/d. We assessed the effectiveness of the FA flour fortification program on bread folate content and on blood folate concentration in women of childbearing age in Santiago, Chile. The prefortification folate status of 751 healthy women of reproductive age was assessed. The folate content of 100 bread samples bought at retail bakeries was measured, average wheat flour consumption was estimated and postfortification FA dietary intake was calculated. The effect of flour fortification on blood folate concentration in this group of women (n = 605) was evaluated in a follow-up study. Blood folate concentrations of the 605 women in the follow-up group increased (P < 0.0001) following fortification. Before fortification the mean serum and red blood cell folate concentrations were 9.7 +/- 4.3 and 290 +/- 102 nmol/L, respectively, compared with 37.2 +/- 9.5 and 707 +/- 179 nmol/L postfortification, respectively. The mean FA content of bread was 2020 +/- 940 micro g/kg. The median FA intake of the group evaluated postfortification was 427 microg/d (95% CI 409-445) based on an estimated intake of 219 g/d (95% CI 201-229) of wheat flour, mainly as bread. Fortification of wheat flour substantially improved folate status in a population of women of reproductive age in Chile. The effect of the FA fortification program on the occurrence of NTD is currently being assessed.


Current Chemistry Investigators (CCI): Development and Evaluation of a Scientist in a Classroom Electrochemistry Workshop.
Over the course of the COVID-19 pandemic, school students suffered from a reduction in opportunities to connect with higher education institutions, meet scientific role models in person, discuss scientific career options, and carry out hands-on practical laboratory activities. Current Chemistry Investigators (CCI) is a successful electrochemistry-based STEM career intervention program, developed and evaluated through a co-creation process with teachers and students. The goals of CCI are 2-fold: first, to provide school students with career advice through tangible scientific role models and, second, to provide real-world context for the fundamentals of electrochemistry through hands-on activities. Herein, the development of a novel electro-analytical workshop from concept through to delivery with over a thousand students having taken part to date is reported. Students are tasked with solving why a battery malfunctioned through quantitative and qualitative analyses of an electrolyte using conductivity meters. Student feedback is also gathered anonymously through the use of a classroom response system (also known as "clickers"). Together with feedback from teachers, a robust evaluation is presented to measure the impact of providing tangible scientific role models and the usefulness of the workshop.


Pseudopeptide foldamers: the homo-oligomers of pyroglutamic acid.
As a part of a program evaluating substituted gamma-lactams as conformationally constrained building blocks of pseudopeptide foldamers, we synthesized the homo-oligomers of L-pyroglutamic acid up to the tetramer level by solution methods. The preferred conformation of this pseudopeptide series in structure-supporting solvents was assessed by FT-IR absorption, 1H NMR and CD techniques. In addition, the crystal structure of the N alpha-protected dimer was established by X-ray diffraction. A high-level DFT computational modeling was performed based on the crystallographic parameters. In this analysis, we demonstrated that an alpha C-H...O=C intramolecular hydrogen bond is responsible for the stabilization of the s-trans L-pGlu-L-pGlu conformation by 1.4 kcal mol-1. This effect can be easily detected by 1H NMR spectroscopy, owing to the anomalous chemical shifts of the alpha CH protons present in all of the oligomers. In summary, we have developed a new polyimide-based, foldameric structure that, if appropriately functionalized, has promise as a rigid scaffold for novel functions and applications.


Pulmonary and systemic fungal infections in an Atlantic spotted dolphin and a Bryde's whale, Brazil.
We report the gross and microscopic findings and molecular identification of 2 cases of hyphate fungal infection in cetaceans from Brazil. The first case involved an adult male Atlantic spotted dolphin Stenella frontalis with localized pulmonary disease characterized by pyogranulomatous and necrotizing bronchopneumonia with intralesional hyphae. The second case involved an adult male Bryde's whale Balaenoptera edeni with orchitis, periorchitis, mesenteric lymphadenitis and pyogranulomatous bronchopneumonia with intralesional hyphae. PCR analysis from the dolphin's lung yielded Aspergillus fumigatus, and the fungus from the whale's mesenteric lymph node showed the greatest identity to Nanniziopsis obscura and Stagonosporopsis cucurbitacearum These cases represent the first reports of pulmonary aspergillosis by A. fumigatus in an Atlantic spotted dolphin and systemic mycosis by a possibly novel Onygenales in marine mammals.


Implementation of a Simplistic Bivalirudin-Warfarin Transition Protocol Is Associated With Improved Achievement of INR Values Within the Therapeutic Range.
Transition from bivalirudin to long-term warfarin therapy is often difficult to execute due to bivalirudin prolongation of the international normalized ratio (INR), and literature to help guide this transition is extremely limited. To assess the transition from bivalirudin to warfarin after implementation of an institution-wide transition protocol. In this retrospective quasiexperimental study, adult patients receiving bivalirudin directly followed by warfarin for nonprocedural systemic anticoagulation were evaluated to determine the frequency of successful transition to warfarin. Participants were compared before (preprotocol) and after (postprotocol) the implementation of the transition protocol. A total of 39 patients met inclusion criteria and were included in the analysis (preprotocol = 19; postprotocol = 20). The percentage of patients achieving a successful transition was significantly higher in the postprotocol group compared with the preprotocol group (80.0% vs 42.1%, P = 0.015). Bleeding events were similar between the 2 groups (23.1% vs 16.7%, P = 0.689). Withholding of warfarin doses or the use of anticoagulant reversal agents or blood transfusions for supratherapeutic INR levels, surgical procedures, or drop in hemoglobin was numerically lower in the postprotocol group compared with the preprotocol group (16.7% vs 46.2%, P = 0.202). Implementation of a simplistic bivalirudin-warfarin transition protocol significantly increased the frequency of therapeutic INR results on bivalirudin discontinuation. Additionally, patients treated according to this protocol were less likely to have warfarin doses withheld or require reversal agents. Larger studies testing this transition strategy are warranted.


Bone Marrow Culture-Derived Conditioned Medium Recovers Endothelial Function of Vascular Grafts following In Vitro Ischemia/Reperfusion Injury in Diabetic Rats.
Ischemia/reperfusion injury (IRI) remains a challenge in coronary artery bypass grafting (CABG). Diabetic patients with coronary artery disease are more likely to require CABG and therefore run a high risk for cardiovascular complications. Conditioned medium (CM) from bone marrow-derived mesenchymal stem cells has been shown to have beneficial effects against IRI. We hypothesized that adding CM to physiological saline protects vascular grafts from IRI in diabetic rats. Bone-marrow derived cells were isolated from nondiabetic rat femurs/tibias, and CM was generated. As we previously reported, CM contains 23 factors involved in inflammation, oxidative stress, and apoptosis. DM was induced by streptozotocin administration. Eight weeks later, to measure vascular function, aortic rings were isolated and mounted in organ bath chambers (DM group) or stored in 4°C saline, supplemented either with a vehicle (DM-IR group) or CM (DM-IR+CM group). Although DM was associated with structural changes compared to controls, there were no functional alterations. However, compared to the DM group, in the DM-IR aortas, impaired maximum endothelium-dependent vasorelaxation in response to acetylcholine (DM 86.7 ± 0.1% vs. DM-IR 42.5 ± 2.5% vs. DM-IR+CM 61.9 ± 2.0%, p < 0.05) was improved, caspase-3, caspase-8, caspase-9, and caspase-12 immunoreactivity was decreased, and DNA strand breakage, detected by the TUNEL assay, was reduced by CM. We present the experimental finding that the preservation of vascular grafts with CM prevents endothelial dysfunction after IRI in diabetic rats. Targeting apoptosis by CM may contribute to its protective effect.


The advanced glycation end product N(epsilon)-carboxymethyllysine is not a predictor of cardiovascular events and renal outcomes in patients with type 2 diabetic kidney disease and hypertension.
Advanced glycation end products (AGEs) are implicated in the pathogenesis of vascular damage, especially in patients with diabetes and renal insufficiency. The oxidatively formed AGE N(epsilon)-carboxymethyllysine (CML) is thought to be a marker of oxidative stress. Four hundred fifty patients with type 2 diabetes and nephropathy from the Irbesartan in Diabetic Nephropathy Trial cohort (mean age, 58 +/- 8.2 years; 137 women, 313 men) with a mean glomerular filtration rate of 48.2 mL/min (0.80 mL/s; Modification of Diet in Renal Disease formula) were followed up for 2.6 years. Serum CML was measured by using an enzyme-linked immunosorbent assay. Relationships between CML levels, traditional risk factors, and cardiovascular and renal events were tested in Cox proportional hazards models. Mean serum CML level was 599.9 +/- 276.0 ng/mL, and mean hemoglobin A1c level was 7.5% +/- 1.6%. One hundred forty-three first cardiovascular events occurred during follow-up; 74 patients died, 44 of cardiovascular causes. Final multivariate analysis showed age (relative risk [RR], 1.87; confidence interval [CI], 1.13 to 3.11; P = 0.016 for the highest compared with lowest quartile), history of prior cardiovascular events (RR, 1.96; CI, 1.35 to 2.85; P < 0.0005), and 24-hour urinary albumin-creatinine ratio (RR, 1.29; CI, 1.11 to 1.50 per doubling; P < 0.0005) to be independent risk factors for a first cardiovascular event, but not CML level. CML level also did not correlate significantly with renal outcome. Serum CML level could not be identified as an independent risk factor for cardiovascular or renal outcomes in the examined population. This suggests that traditional risk factors might have a more important role for these end points or that other AGE compounds, as well as tissue AGE levels, might be of greater relevance compared with serum levels, which remains open to further study.


Antigenic properties of cystatin-binding cysteine proteinases from Neodiplostomum seoulense.
The antigenic properties of cysteine proteinases binding to cystatin were analyzed in Neodiplostomum seoulense, an intestinal trematode that infects humans and rodents in the Republic of Korea. Cystatin was found to effectively capture cysteine proteinases present in the crude extract of N. seoulense. The IgG levels against cystatin-binding cysteine proteinases in sera of mice infected with N. seoulense were higher than those in sera of mice immunized with the crude extract of N. seoulense. The production of IgG antibodies against cystatin-binding cysteine proteinases increased according to the length of infection period. In immunoblots of purified cystatin-binding proteinases, 2 molecules, approximately 50 kDa and 60 kDa, reacted with N. seoulense-infected mouse sera. Of the sera from patients infected with various helminths, those of sparganum-infected patients showed the strongest affinities for cystatin-binding cysteine proteinases of N. seoulense. Cystatin-binding cysteine proteinases of N. seoulense are suggested to be putative antigens for serodiagnosis of human N. seoulense infection.


Kinetic Modeling of API Oxidation: (1) The AIBN/H2O/CH3OH Radical "Soup".
Stress testing of active pharmaceutical ingredients (API) is an important tool used to gauge chemical stability and identify potential degradation products. While different flavors of API stress testing systems have been used in experimental investigations for decades, the detailed kinetics of such systems as well as the chemical composition of prominent reactive species, specifically reactive oxygen species, are unknown. As a first step toward understanding and modeling API oxidation in stress testing, we investigated a typical radical "soup" solution an API is subject to during stress testing. Here we applied ab initio electronic structure calculations to automatically generate and refine a detailed chemical kinetics model, taking a fresh look at API oxidation. We generated a detailed kinetic model for a representative azobis(isobutyronitrile) (AIBN)/H2O/CH3OH stress-testing system with a varied cosolvent ratio (50%/50%-99.5%/0.5% vol water/methanol) for 5.0 mM AIBN and representative pH values of 4-10 at 40 °C that was stirred and open to the atmosphere. At acidic conditions, hydroxymethyl alkoxyl is the dominant alkoxyl radical, and at basic conditions, for most studied initial methanol concentrations, cyanoisopropyl alkoxyl becomes the dominant alkoxyl radical, albeit at an overall lower concentration. At acidic conditions, the levels of cyanoisopropyl peroxyl, hydroxymethyl peroxyl, and hydroperoxyl radicals are relatively high and comparable, while, at both neutral and basic pH conditions, superoxide becomes the prominent radical in the system. The present work reveals the prominent species in a common model API stress testing system at various cosolvent and pH conditions, sets the stage for an in-depth quantitative API kinetic study, and demonstrates the usage of novel software tools for automated chemical kinetic model generation and ab initio refinement.