Assessment of antioxidant activity of extracts from unique Greek varieties of Leguminosae plants using in vitro assays.
It is believed that legumes are a very good source of micronutrients and phytochemicals that present chemopreventive activity against diseases such as diabetes, coronary heart disease and colon cancer. Methanolic and aqueous extracts from 11 unique varieties of Leguminosae family plants cultured in Greece were tested using three different in vitro assays in order to investigate the mechanisms by which phytochemicals present in these legumes exert their chemoprevention. The extracts were tested by the 1, -diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, the hydroxyl radical- and the peroxyl radical-induced DNA strand scission assays. Hydroxyl (OH*) and peroxyl (ROO*) radicals were generated from ultraviolet (UV) photolysis of hydrogen peroxide (H2O2) and thermal decomposition of 2,2'-azobis-(2-amidinopropane hydrochloride) (AAPH) respectively. In the DPPH assay, all the tested extracts displayed potent radical scavenging efficiency. Furthermore, most of the Leguminosae family plant extracts exerted significant protective activity against DNA damage induced by both reactive oxygen species, although they were more effective in inhibiting ROO*-induced rather than OH*-induced DNA strand scission. The results suggest that the free radical scavenging activity of Leguminosae plants may be one of the mechanisms accounting for their chemoprevention.


Rapid killing of Mycobacterium terrae by N-chlorotaurine in the presence of ammonium is caused by the reaction product monochloramine.
We have studied the activity of the weak endogenous oxidant N-chlorotaurine against Mycobacterium terrae. The study revealed slow killing of more than 2h duration by 1% (55 mM) N-chlorotaurine. In the presence of ammonium chloride, however, killing times decreased to a few minutes, even by 0.1% N-chlorotaurine. This phenomenon is explained by formation of the lipophilic and therefore more bactericidal monochloramine as a result of transhalogenation of ammonia by N-chlorotaurine.


Taurine and skeletal muscle disorders.
Taurine is abundantly present in skeletal muscle. We give evidence that this amino acid exerts both short-term and long-term actions in the control of ion channel function and calcium homeostasis in striated fibers. Short-term actions can be estimated as the ability of this amino acid to acutely modulate both ion channel gating and the function of the structures involved in calcium handling. Long-term effects can be disclosed in situations of tissue taurine depletion and are likely related to the ability of the intracellular taurine to control transducing pathways as well as homeostatic and osmotic equilibrium in the tissue. The two activities are strictly linked because the intracellular level of taurine modulates the sensitivity of skeletal muscle to the exogenous application of taurine. Myopathies in which ion channels are directly or indirectly involved, as well as inherited or acquired pathologies characterized by metabolic alterations and change in calcium homeostasis, are often correlated with change in muscle taurine concentration and consequently with an enhanced therapeutic activity of this amino acid. We discuss both in vivo and in vitro evidence that taurine, through its ability to control sarcolemmal excitability and muscle contractility, can prove beneficial effects in many muscle dysfunctions.


Identity of acyl-CoA oxidase with glutaryl-CoA oxidase.
Rats fed on clofibrate- and DEHP-containing diets showed virtually proportional increases in hepatic acyl-CoA oxidase and glutaryl-CoA oxidase activities. The solubilization profiles of the two activities from the light mitochondrial fraction of the liver homogenate of DEHP-treated rats were the same, and the glutaryl-CoA oxidase/acyl-CoA oxidase activity ratio remained constant through the purification. The final preparation obtained was a single protein based on the result of polyacrylamide gel electrophoresis. The evidence indicates that the two activities are associated with the same protein.


The implementation of the Kinyoun staining technique in a resource-limited setting is feasible and reveals a high prevalence of intestinal cryptosporidiosis in patients with HIV.
In resource-limited settings, intestinal Cryptosporidial or coccidian infections are common causes of chronic diarrhea but usually remain undiagnosed by routine stool investigation. Here, the addition of the Kinyoun staining technique after stool concentration was evaluated as an easy and inexpensive method for diagnosis of intestinal parasitic infection in patients with HIV. This cross-sectional study investigated patients with HIV with diarrhea and randomly selected patients with HIV without diarrhea as controls. Stool samples were examined by wet mount microscopy and Kinyoun staining after stool concentration. Clinical, sociodemographic, and behavioral data were collected. Statistical analysis was performed using chi-squared test and multivariate regression analysis. In total, 163 participants were included (62.0% female, mean age 38.2 [SD ± 10.7] years). Diarrhea was present in 52.1% (85/163). The prevalence of intestinal parasites was 18.4% (30/163). Cryptosporidial infections were more frequent among patients with diarrhea (12.9% [11/85] vs 1.3% [1/78], P = 0.005) and in patients with CD4+ cell count <200 cells/µl (25.9% [7/27] vs 3.7% [5/136], P = 0.001). Risk factors for intestinal parasitic infections were diarrhea and the habit of regularly eating uncooked food. Kinyoun staining was necessary for the detection of cryptosporidiosis. In our cohort, the prevalence of intestinal parasitic infection was high, especially after additional use of Kinyoun staining for detection of Cryptosporidia or intestinal coccidia. Considering its clinical relevance, particularly in individuals at risk, the implementation of this technique should be considered in resource-limited settings.


Hibernations of electroactive bacteria provide insights into the flexible and robust BOD detection using microbial fuel cell-based biosensors.
Microbial fuel cell (MFC) biosensors have been suggested as an alternative detection method for biochemical oxygen demand (BOD). However, it is absolutely essential to develop maintenance procedures for MFC biosensors` because in practice the lay-up period cannot be avoided. In this work, setting electroactive bacteria (EAB) under hibernation condition was demonstrated to be a feasible maintenance method, which provided important insights into the flexible and robust BOD detection using MFC biosensors. Standard BOD solution containing 500, 200, and 20 mg/L BOD were used to evaluate the detection performance after EAB hibernations. Results demonstrated quick recovery of voltage output and high-accuracy BOD detection after hibernations up to 30 days in MFC biosensors detecting 500 mg/L and 200 mg/L BOD. Identical anode potentials after the EAB hibernations suggested intact bacterial ability of current generation. Non-turnover cyclic voltammetry immediately collected after the hibernations suggested multiple redox couples and the presence of cytochromes that played key roles in EAB metabolism and functioned as temporary electron sinks during the hibernations, leading to the increased detected BOD concentration in the restarting cycles. Generally, setting EAB under hibernation condition is a simple and convenient maintenance method for MFC-based BOD biosensors, which not only provides insights into flexible and robust BOD detection, but also be helpful for other MFC biosensing instruments.


[Influence of the degree of aeration on halotolerance of yeasts of the genera Candida, Rhodotorula, and Malassezia].
The biochemical mechanisms were studied that determine different reactions of yeasts of different genera to two simultaneously imposed stressors, hypoxia and osmotic shock. For Candida lipolytica, these two stressors were antagonistic, which resulted in stimulation of yeast growth by NaCl (in a wide range of concentrations) under microaerobic conditions. The reaction of Malassezia sp. was different: the degree of halotolerance of this microorganism was lower under microaerobic conditions. An intervening reaction pattern was characteristic of Rhodotorula aurantiaca. These differences were found to be determined, above all, by the induction of a salt-resistant respiratory system (oxidase) in Candida lipolytica, which could not be induced in Malassezia sp. In addition, the synthesis of catalase was enhanced in Candida lipolytica, which provided for neutralization of the active forms of oxygen accumulating as a result of inhibition of other protective enzymes by salt.


De novo generation of infectious prions with bacterially expressed recombinant prion protein.
The prion hypothesis is strongly supported by the fact that prion infectivity and the pathogenic conformer of prion protein (PrP) are simultaneously propagated in vitro by the serial protein misfolding cyclic amplification (sPMCA). However, due to sPMCA's enormous amplification power, whether an infectious prion can be formed de novo with bacterially expressed recombinant PrP (rPrP) remains to be satisfactorily resolved. To address this question, we performed unseeded sPMCA with rPrP in a laboratory that has never been exposed to any native prions. Two types of proteinase K (PK)-resistant and self-perpetuating recombinant PrP conformers (rPrP-res) with PK-resistant cores of 17 or 14 kDa were generated. A bioassay revealed that rPrP-res(17kDa) was highly infectious, causing prion disease in wild-type mice with an average survival time of about 172 d. In contrast, rPrP-res(14kDa) completely failed to induce any disease. Our findings reveal that sPMCA is sufficient to initiate various self-perpetuating PK-resistant rPrP conformers, but not all of them possess in vivo infectivity. Moreover, generating an infectious prion in a prion-free environment establishes that an infectious prion can be formed de novo with bacterially expressed rPrP.


Partition level multiview subspace clustering.
Multiview clustering has gained increasing attention recently due to its ability to deal with multiple sources (views) data and explore complementary information between different views. Among various methods, multiview subspace clustering methods provide encouraging performance. They mainly integrate the multiview information in the space where the data points lie. Hence, their performance may be deteriorated because of noises existing in each individual view or inconsistent between heterogeneous features. For multiview clustering, the basic premise is that there exists a shared partition among all views. Therefore, the natural space for multiview clustering should be all partitions. Orthogonal to existing methods, we propose to fuse multiview information in partition level following two intuitive assumptions: (i) each partition is a perturbation of the consensus clustering; (ii) the partition that is close to the consensus clustering should be assigned a large weight. Finally, we propose a unified multiview subspace clustering model which incorporates the graph learning from each view, the generation of basic partitions, and the fusion of consensus partition. These three components are seamlessly integrated and can be iteratively boosted by each other towards an overall optimal solution. Experiments on four benchmark datasets demonstrate the efficacy of our approach against the state-of-the-art techniques.


Impact of Social Vulnerability on Access to Educational Programming Designed to Enhance Living Donation.
Transplant candidate participation in the Living Donor Navigator Program is associated with an increased likelihood of achieving living donor kidney transplantation; yet not every transplant candidate participates in navigator programming. We sought to assess interest and ability to participate in the Living Donor Navigator Program by the degree of social vulnerability. Eighty-two adult kidney-only candidates initiating evaluation at our center provided Likert-scaled responses to survey questions on interest and ability to participate in the Living Donor Navigator Program. Surveys were linked at the participant-level to the Centers for Disease Control and Prevention Social Vulnerability Index and county health rankings and overall social vulnerability and subthemes, individual barriers, telehealth capabilities/ knowledge, interest, and ability to participate were assessed utilizing nonparametric Wilcoxon ranks sums tests, chi-square, and Fisher's exact tests. Participants indicating distance as a barrier to participation in navigator programming lived approximately 82 miles farther from our center. Disinterested participants lived in areas with the highest social vulnerability, higher physical inactivity rates, lower college education rates, and higher uninsurance (lack of insurance) and unemployment rates. Similarly, participants without a computer, who never heard of telehealth, and who were not encouraged to participate in telehealth resided in areas of highest social vulnerability. These data suggest geography combined with being from under-resourced areas with high social vulnerability was negatively associated with health care engagement. Geography and poverty may be surrogates for lower health literacy and fewer health care interactions.


A supercritical fluid technology for liposome production and comparison with the film hydration method.
Liposomes were produced by an innovative method using supercritical carbon dioxide as a dispersing agent. A quality by design strategy was used to find optimal production conditions with specific parameters (lipid concentration, dispersion volume, agitation rate, temperature and pressure) allowing the production of liposomes with predicted physicochemical characteristics (particles size and PdI). Two conditions were determined with specific production parameters. It was shown that these two conditions allowed the production of liposomes of different compositions and that most of the liposome formulations had size and dispersity in accordance with the prediction values. The condition involving the higher lipid concentration showed a higher variability in terms of size and dispersity. However, this variability remained acceptable. This innovative supercritical method allowed the production of liposomes with physicochemical characteristics similar to those obtained by the conventional thin film hydration method. This new supercritical carbon dioxide method easily scalable in GMP conditions is a one-step production method contrarily to conventional methods which generally need an additional step as extrusion to homogenize the size of liposomes.


Serum uric acid and risk for development of hypertension and impaired fasting glucose or Type II diabetes in Japanese male office workers.
We examined the association of serum uric acid (SUA) with development of hypertension (blood pressure > or = 140/90 mmHg and/or medication for hypertension) and impaired fasting glucose (IFG) (a fasting plasma glucose level 6.1-6.9 mmol/l) or Type II (non-insulin-dependent) diabetes (a fasting plasma glucose level > or = 7.0 mmol/l and/or medication for diabetes) over a 6-year follow-up among 2310 Japanese male office workers aged 35-59 years who did not have hypertension, IFG, Type II diabetes, or past history of cardiovascular disease at study entry. After controlling for potential predictors of hypertension and diabetes, the relative risk for hypertension compared with quintile 1 of SUA level was 1.27 [95% confidence interval (CI): 1.00-1.62] for quintile 2, 1.34 (95% CI: 1.08-1.74) for quintile 3, 1.48 (95% CI: 1.18-1.89) for quintile 4, and 1.58 (95% CI: 1.26-1.99) for quintile 5 (p for trend <0.001). The respective multivariate-adjusted relative risks for IFG or Type II diabetes compared with quintile 1 of SUA level were 1.55 (95% CI: 0.95-2.63), 1.62 (95% CI: 0.98-2.67), 1.61 (95% CI: 1.01-2.58), and 1.78 (95% CI: 1.11-2.85) (p for trend = 0.030). The association between SUA level and risk for hypertension and IFG or Type II diabetes was stronger among men with a body mass index (BMI) <24.2 kg/m2 than among men with a BMI > or = 24.2 kg/m2, although the absolute risk was greater in more obese men. These results indicate that SUA level is closely associated with an increased risk for hypertension and IFG or Type II diabetes.


Multicentre phase II trial of trastuzumab and capecitabine in patients with HER2 overexpressing metastatic pancreatic cancer.
New therapeutic options for metastatic pancreatic cancer are urgently needed. In pancreatic cancer, overexpression of the epidermal growth factor receptor 2 (HER2) has been reported in up to 45%. This multicentre phase II study investigated the efficacy and toxicity of the HER2 antibody trastuzumab combined with capecitabine in the patients with pancreatic cancer and HER2 overexpression. Primary endpoint was progression-free survival (PFS) after 12 weeks. A total of 212 patients were screened for HER2 expression. Immunohistochemical (IHC) HER2 expression was: 83 (40%) grade 0, 71 (34%) grade 1, 31 (15%) grade 2, 22 (11%) grade 3. A total of 17 patients with IHC +3 HER2 expression or gene amplification could be assessed for the treatment response. Grade 3/4 treatment toxicities were: each 7% leucopenia, diarrhoea, nausea and hand-foot syndrome. Progression-free survival after 12 weeks was 23.5%, median overall survival (OS) 6.9 months. This study demonstrates +3 HER2 expression or gene amplification in 11% of patients. Contrary to breast and gastric cancer, only 7 out of 11 (64%) patients with IHC +3 HER2 expression showed gene amplification. Although the therapy was well tolerated, PFS and OS did not perform favourably compared with standard chemotherapy. Together, we do not recommend further evaluation of anti-HER2 treatment in patients with metastatic pancreatic cancer.


BDNF mobilizes synaptic vesicles and enhances synapse formation by disrupting cadherin-beta-catenin interactions.
Neurons of the vertebrate central nervous system have the capacity to modify synapse number, morphology, and efficacy in response to activity. Some of these functions can be attributed to activity-induced synthesis and secretion of the neurotrophin brain-derived neurotrophic factor (BDNF); however, the molecular mechanisms by which BDNF mediates these events are still not well understood. Using time-lapse confocal analysis, we show that BDNF mobilizes synaptic vesicles at existing synapses, resulting in small clusters of synaptic vesicles "splitting" away from synaptic sites. We demonstrate that BDNF's ability to mobilize synaptic vesicle clusters depends on the dissociation of cadherin-beta-catenin adhesion complexes that occurs after tyrosine phosphorylation of beta-catenin. Artificially maintaining cadherin-beta-catenin complexes in the presence of BDNF abolishes the BDNF-mediated enhancement of synaptic vesicle mobility, as well as the longer-term BDNF-mediated increase in synapse number. Together, this data demonstrates that the disruption of cadherin-beta-catenin complexes is an important molecular event through which BDNF increases synapse density in cultured hippocampal neurons.


Age-specific incidence and prevalence rates of treated epilepsy in an unselected population of 2,052,922 and age-specific fertility rates of women with epilepsy.
There are no data on prevalence or incidence of treated epilepsy, and no data on fertility of women with epilepsy from an unselected UK population. We used the General Practice Research Database to ascertain the incidence and prevalence of people with treated epilepsy in an unselected population of 2,052,922 people in England and Wales, and also age-specific fertility rates. We defined period prevalence of treated epilepsy as the number of people with epilepsy taking an antiepileptic drug per 100,000 people during 1995. The incidence of treated epilepsy was defined as the number of new cases of treated epilepsy per 100,000 people during the same period. We calculated fertility rates among women with treated epilepsy between 1991 and 1995 and compared these rates with the population rates for England and Wales in 1993. The period prevalence of treated epilepsy in 1995 was 5.15 per 1000 people (95% CI 5.05-5.25). The prevalence was lower in children (age 5-9 years 3.16 [2.86-3.48]; 10-14 years 4.05 [3.70-4.42]), and higher in older people (65-69 years 6.01 [5.50-6.57]; 70-74 years 6.53 [5.97-7.14]; 75-79 years 7.39 [6.73-8.11]); 80-84 years 7.54 [6.78-8.39]; 85 years and older 7.73 [6.98-8.66]). The incidence of treated epilepsy was 80.8 per 100,000 people (76.9-84.7). The incidence was lower in children (5-9 years 63.2 [50.5-79.1]; 10-14 years 53.8 [42.4-68.3]) and higher in older people (65-69 years 85.9 [68.5-107.3]; 70-74 years 82.8 [65.0-105.2]; 75-79 years 114.5 [116.9-179.2]; 80-84 years 159 [125.2-202.6]; > or = 85 years 135.4 [100.4-178.7]). Fertility was lower among women with treated epilepsy, with an overall rate of 47.1 livebirths per 1000 women aged 15-44 per year (42.3-52.2), compared with a national rate of 62.6 in the same age-group. The standardised fertility ratios were significantly lower between the ages of 25 and 39 years in women with epilepsy (p<0.001). Compared with previous studies, we found that the incidence of epilepsy was higher in elderly people and lower in children. The prevalence rates also increase with age. Women aged 25-39 years with treated epilepsy have significantly lower fertility rates than those in the general population. Research is needed to identify any potentially preventable causes for the low fertility rates.


Huge left atrial appendage aneurysm revealed by chronic hiccups.
Left atrial appendage (LAA) aneurysm is an extremely rare anomaly. So far, less than one hundred cases only have been reported worldwide. Revelation modes are dominated by complications such as arrhythmias and thromboembolic events. We herein report a pediatric case of huge congenital LAA aneurysm with an original revelation mode that has never been described before in medical literature.


Expression of H-2b alloantigens in a variant of a Moloney virus-induced YAC (H-2a) lymphoma.
Splenocytes of A(H-2a), BALB/c(H-2d) and C57BL/6(H-2b) mice that were sensitized against several allogeneic normal lymphocytes and lymphoma cells in mixed lymphocyte cultures (MLC) and mixed lymphocyte-tumor cell cultures (MLTC) exhibited the expected cell-mediated cytotoxic responses against normal and tumor cells carrying the relevant alloantigens. In contrast, strong cytotoxic reactivity cross-reactive with C57BL/6 target cells was observed when a variant of YAC lymphoma (YAC-b) of strain A mice were employed either as sensitizers or as targets. Furthermore, this 'unpredicted' cross-reactivity was also obtained with syngeneic responder splenocytes sensitized to YAC-b cells. The possibility that the nonspecific cytotoxicity was caused by activation of natural killer (NK) cells in the culture was excluded by the finding that the cytotoxic capability of sensitized effector cells was greatly diminished by treatment with anti-Thy 1.2 serum and complement. Serologic analysis with anti-H-2 alloantisera revealed that the YAC-b tumor line, but not the parental YAC lymphoma, carried H-2b alloantigens. It is concluded that the 'unpredicted' cytotoxic responses here observed is a result of conventional antigen recognition and of specific sensitization to alien H-2b components expressed on the YAC-b subline.


A comparison of colorimetric and clonogenic assays for hypoxic-specific toxins with hamster and human cells.
The hypoxic cytotoxicities of misonidazole and pimonidazole (Ro 03-8799) towards the human tumor cell lines HT-1080 and LoVo have been compared with those seen with Chinese hamster V79-379A cells. Survival was assayed using two colorimetric assays, either a tetrazolium salt (MTT) or methylene blue, and by conventional colony scoring. The drugs were more cytotoxic towards HT-1080 and LoVo cells than V79 cells. The times taken for 10 mmol dm-3 misonidazole to reduce survival to 0.1 surviving fraction (SF) using colony formation as the end point were 2.6 hr for HT-1080, 2.4 hr for LoVo, and 3.5 hr for V79; using the MTT assay these times were 3.5 hr, 2.1 hr, and 2.9 hr, respectively. The times for 2 mmol dm-3 pimonidazole to reduce survival to 0.1 SF using colony formation as the end point were 2.0 hr for HT-1080, 1.7 hr for LoVo, and 3.7 hr for V79; using the MTT assay these times were 2.5 hr, 1.4 hr, and 2.5 hr, respectively.


Determinants of 90-day readmission following ventral hernia repair with and without myocutaneous flap reconstruction: a National Readmissions Database analysis.
Readmission status is an important clinical component of healthcare outcomes. 90-day readmission following complex open ventral hernia repair has not been well studied with national level data. This study aims to compare readmission rates for patients undergoing standard vs. complex (myocutaneous flap-based) ventral hernia repair. We hypothesize that complexity of reconstruction will be an independent predictor of readmission after ventral hernia repair. A retrospective cohort study was performed with 1:1 matching of hernia repair type using the National Readmissions Database. Patients were selected using ICD-9 codes corresponding to ventral hernia repair with or without myocutaneous flap. 90-day readmissions were determined on patients within the first through third quarters of each year. After matching, a multivariable logistic regression analysis was performed using confounding variables including hospital setting, comorbidities, type of repair, urgency of repair, sociodemographic status, and payer. Likelihood of 90-day readmission was calculated from odds ratios. Readmission rates were 19.1% (38,313 out of 200,266) and 22.5% (692 out of 3075) at 90-day for standard ventral hernia repair and complex ventral hernia repair, respectively. 3116 standard ventral hernia repair patients were matched with 3074 complex ventral hernia repair patients. After matching there was a significantly increased readmission rate for repairs involving myocutaneous flaps, with odds ratio (OR) 1.30 (95% CI 1.22-1.60). Payer status (OR 1.82; 95% CI 1.21-2.74), teaching hospital status (OR 1.42; 95% CI 1.23-1.64) and income quartile (OR 1.35; CI 1.10-1.65) were independent predictors of readmission. Patients undergoing myocutaneous flap-based reconstruction have higher readmission rates than those undergoing less complex ventral hernia repair. Socioeconomic disparity as reflected in payer status is a particularly strong predictor of readmission. The data support the concept that focused efforts are needed to optimize patient outcomes for patients requiring more complex repair, including socioeconomically disadvantaged patient populations.


The nurse's role in bowel awareness.
Promoting good health and preventing ill health are part of the nurse's role, and using a model for health promotion in the colorectal nursing service is an effective way to improve patient care. This article considers referral guidelines for patients with suspected colorectal cancer and suggests methods of raising public awareness.


Submicromolar Ca2+ regulates phosphorylating respiration by normal rat liver and AS-30D hepatoma mitochondria by different mechanisms.
The stimulation of 2-oxoglutarate and NAD(+)-isocitrate dehydrogenase by Ca2+ in mitochondria from normal tissues has been proposed to mediate partially the activation of oxidative energy metabolism elicited by physiological elevations in cytosolic Ca2+. This mode of regulation may also occur in tumor cells in which several aspects of mitochondrial metabolism are known to be altered. This study provides a comparison of the stimulation by submicromolar concentrations of Ca2+ on the rates of ATP-generating (state 3) respiration under physiologically realistic conditions by mitochondria isolated from normal rat liver and from highly malignant rat AS-30D ascites hepatoma cells. The K0.5 for activation of glutamate-dependent state 3 respiration by Ca2+ in the presence of ATP at 37 degrees C was determined to be 0.70 +/- 0.05 (S.E.) microM for hepatoma mitochondria and 0.90 +/- 0.03 microM for rat liver mitochondria. This activation was also reflected by a Ca2(+)-induced shift in the oxidation-reduction state of hepatoma mitochondrial pyridine nucleotides to a more reduced level and Ca2+ stimulation of 14CO2 production from [1-14C]glutamate. Whereas the Ca2+ sensitivity of state 3 respiration by hepatoma mitochondria can be explained by the activation of 2-oxoglutarate and possibly NAD(+)-isocitrate dehydrogenases, the Ca2+ sensitivity of liver mitochondrial respiration appears to be predominantly mediated by activation of electron flow through ubiquinone and Complex III of the electron transport chain, as indicated by the specificity of the effects of Ca2+ on respiration with different oxidizable substrates. Although rat liver and hepatoma mitochondria employ different modes of Ca2(+)-activated ATP generation, these results support the hypothesis that changes in cytosolic Ca2+ play a significant role in the potentiation of energy production in tumor, as well as normal tissue.


Efficient embryogenic suspension culturing and rapid transformation of a range of elite genotypes of sweet potato (Ipomoea batatas [L.] Lam.).
Efficient Agrobacterium tumefaciens-mediated transformation was developed using embryogenic suspension cell cultures of elite sweet potato (Ipomoea batatas [L.] Lam.) cultivars, including Ayamurasaki, Sushu2, Sushu9, Sushu11, Wanshu1, Xushu18 and Xushu22. Embryogenic suspension cultures were established in LCP medium using embryogenic calli induced from apical or axillary buds on an induction medium containing 2 mg l(-1) 2,4-D. Suspension cultures were co-cultivated with A. tumefaciens strain LBA4404 harboring the binary plasmid pCAMBIA1301 with the hpt gene as a selectable marker and an intron-interrupted uidA gene as a visible marker. Several key steps of the sweet potato transformation system have been investigated and optimized, including the appropriate antibiotics and their concentrations for suppressing Agrobacterium growth and the optimal doses of hygromycin for transformant selection. A total of 485 putative transgenic plant lines were produced from the transformed calli via somatic embryogenesis and germination to plants under 10 mg l(-1) hygromycin and 200 mg l(-1) cefotaxime. PCR, GUS and Southern blot analyses of the regenerated plants showed that 92.35% of them were transgenic. The number of T-DNA insertions varied from one to three in most transgenic plant lines. Plants showed 100% survival when 308 transgenics were transferred to soil in the greenhouse and then to the field. Most of them were morphologically normal, with the production of storage roots after 3 months of cultivation in the greenhouse or fields. The development of such a robust transformation method suitable to a range of sweet potato genotypes not only provides a routine tool for genetic improvement via transgenesis but also allows us to conduct a functional verification of endogenous genes in sweet potato.


Reversible immobilization of invertase on Cu-chelated polyvinylimidazole-grafted iron oxide nanoparticles.
Polyvinylimidazole (PVI)-grafted iron oxide nanoparticles (PVIgMNP) were prepared by grafting of telomere of PVI on the iron oxide nanoparticles. Different metal ions (Cu(2+), Zn(2+), Cr(2+), Ni(2+)) ions were chelated on polyvinylimidazole-grafted iron oxide nanoparticles, and then the metal-chelated magnetic particles were used in the adsorption of invertase. The maximum invertase immobilization capacity of the PVIgMNP-Cu(2+) beads was observed to be 142.856 mg/g (invertase/PVIgMNP) at pH 5.0. The values of the maximum reaction rate (V max) and Michaelis-Menten constant (Km) were determined for the free and immobilized enzymes. The enzyme adsorption-desorption studies, pH effect on the adsorption efficiency, affinity of different metal ions, the kinetic parameters and storage stability of free and immobilized enzymes were evaluated.


The impact of propranolol, 17α-ethinylestradiol, and gemfibrozil on early life stages of marine organisms: effects and risk assessment.
Pharmaceuticals are ubiquitously detected in the marine environment at the ng-μg/L range. Given their biological activity, these compounds are known to induce detrimental effects on biota at relatively low exposure levels; however, whether they affect early life stages of marine species is still unclear. In this study, a set of bioassays was performed to assess the effects of propranolol (PROP), 17-α ethinylestradiol (EE2), and gemfibrozil (GEM) on gamete fertilization and embryonic development of mussels (Mytilus galloprovincialis) and sea urchins (Paracentrotus lividus), and on the survival of seabream (Sparus aurata) larvae. Treatments of PROP (500, 5000, 50,000 ng/L), EE2 (5, 50, 500 ng/L), and GEM (50, 500, 5000 ng/L) were selected to encompass levels comparable or superior to environmental concentrations. Obtained data were tested for dose-response curve fitting and the lowest EC10/LC10 used to calculate risk quotients (RQs) based on the MEC/PNEC. No alteration was induced by PROP on the mussel gamete fertilization, while inhibitory effects were observed at environmental levels of EE2 (500 ng/L) and GEM (5000 ng/L). Fertilization was significantly reduced in sea urchin at all PROP and EE2 dosages. The 48-h exposure to all pharmaceuticals induced the onset of morphological abnormalities in either mussel or sea urchin embryos. Alterations were generally observed at environmentally relevant dosages, except for PROP in mussels, in which alterations occurred only at 50,000 ng/L. A decreased survival of seabream larvae was recorded after 96-h exposure to PROP (all treatments), EE2 (50-500 ng/L), and GEM (500 ng/L). A median RQ > 1 was obtained for all pharmaceuticals, assigning a high risk to their occurrence in marine environments. Overall, results showed that current levels of contamination by pharmaceuticals can impact early stages of marine species, which represent critical junctures in the resilience of coastal ecosystems.


Design and preparation of functional azo linked polymers for the adsorptive removal of bisphenol A from water: Performance and analysis of the mechanism.
In order to effectively remove refractory bisphenol A (BPA) from water, a novel nitrogen doped organic porous functional azo linked polymer (ALP-p) was designed and prepared according to the physicochemical characteristics of propane linked to two phenol hydroxyl groups. This ALP-p was synthesized with 98.5% yield, from pararosaniline and phloroglucinol, via the diazo coupling reaction to produce multiple adsorption functional groups of benzene ring, hydroxyl group and azo group. This functional material showed high adsorption capacity of 357.8 mg/g for 50 mg/L BPA, at 20 °C. The adsorption kinetics and isotherms were described by the pseudo-second-order and Langmuir model, respectively. The major adsorption mechanisms were attributed to the high specific surface area (259.8 m2/g) and pore volume (0.56 cm3/g) related surface adsorption and pore diffusion through porous stereoscopic stacking cavity anchorage. The functional group from the three-dimensional skeleton structures of ALP-p for BPA anchoring endowed chemisorption via π-π interaction between benzene rings and hydrogen-bonding (O-H⋯O, C-H⋯N, C-H⋯O and C-H⋯C) with the hydrogen atom of benzene ring, -OH from BPA and -OH, NN from ALP-p, respectively. The coexisting organic pollutants and alkali environment posed a negative effect on adsorption, while salinity had no significant effect on the process. The adsorption capacity and recovery of ALP-p were >93.5% and 81.6% after five cycles of operation.


Statistical optimization of process parameters for the production of tannase by Aspergillus flavus under submerged fermentation.
Production of tannase by Aspergillus flavus (MTCC 3783) using tamarind seed powder as substrate was studied in submerged fermentation. Plackett-Burman design was applied for the screening of 12 medium nutrients. From the results, the significant nutrients were identified as tannic acid, magnesium sulfate, ferrous sulfate and ammonium sulfate. Further the optimization of process parameters was carried out using response surface methodology (RSM). RSM has been applied for designing of experiments to evaluate the interactive effects through a full 31 factorial design. The optimum conditions were tannic acid concentration, 3.22 %; fermentation period, 96 h; temperature, 35.1 °C; and pH 5.4. Higher value of the regression coefficient (R2 = 0.9638) indicates excellent evaluation of experimental data by second-order polynomial regression model. The RSM revealed that a maximum tannase production of 139.3 U/ml was obtained at the optimum conditions.


Light-scattering and scanning transmission electron microscopic investigation of the hemocyanin of the bivalve, Yoldia limatula (Say).
1. The hemocyanin of the bivalve, Yoldia limatula (Say) was found by light-scattering to have a mol. wt of 8.0 +/- 0.6 x 10(6). Mass measurements by scanning transmission electron microscopy (STEM) gave a particle mass of 8.25 +/- 0.42 x 10(6) for the native particle and 4.09 +/- 0.20 x 10(6) for the half-molecule. 2. The hemocyanin subunits fully dissociated in 8.0 M urea and 6.0 M GdmCl at pH 8.0, and at pH 11.0, 0.01 M EDTA have mol. wts of 4.38 x 10(5), 4.22 x 10(5) and 4.71 x 10(5), close to one-twentieth of the parent molecular weight of Y. limatula hemocyanin and most gastropod hemocyanins. 3. Analyses of the urea dissociation transitions studied at pH 8.0, 1 x 10(-2) M Mg2+, 1 x 10(-2) M Ca2+ and pH 8.0, 3 x 10(-3) M Ca2+ suggest few hydrophobic amino acid groups, of the order of 10 to 15 at the contact areas of each half-molecule or decamer. 4. The further dissociation of the decamers to dimers and the dimers to monomers indicates the presence of a larger number of amino acid groups of ca 35-40/dimer and 100-120/monomer. 5. This suggests hydrophobic stabilization of the dimer to dimer and monomer to monomer contacts within the decamers, as observed with other molluscan hemocyanins.


Beta-amyloid oligomers affect the structure and function of the postsynaptic region: role of the Wnt signaling pathway.
Alzheimer's disease (AD) is the most prevalent neurodegenerative disease in the growing population of elderly people. Synaptic dysfunction is an early manifestation of AD. The cellular mechanism by which beta-amyloid peptide (Abeta) affects synapses remains unclear. Abeta oligomers target synapses in cultured rat hippocampal neurons suggesting that they play a key role in the regulation of synapses. The aim of this work is to study the effect of Abeta oligomers on the central synapses and the possible role of the Wnt signaling pathway in preventing the Abeta effects. We used rat hippocampal neurons, immunofluorescence and western blot procedures to detect synaptic proteins. Abeta oligomers induced a reduction of the postsynaptic density protein 95 (PSD-95) and the NMDA glutamate receptors. We found that Wnt-5a, a noncanonical Wnt ligand, prevents the decrease triggered by Abeta oligomers in the glutamate receptor and PSD-95. Altogether, our results suggest that Abeta oligomers decrease the synaptic responses by affecting the postsynaptic region at different levels. The Wnt signaling activation prevents synaptic damage induced by Abeta, which raises the possibility of a new therapeutic intervention for the treatment of synaptic changes observed in AD.


Memory phenotype CD8+ T cells with innate function selectively develop in the absence of active Itk.
T cells with a memory-like phenotype and possessing innate immune function have been previously identified as CD8(+)CD44(hi) cells. These cells rapidly secrete IFN-gamma upon stimulation with IL-12/IL-18 and are involved in innate responses to infection with Listeria monocytogenes. The signals regulating these cells are unclear. The Tec kinase Itk regulates T cell activation and we report here that a majority of the CD8(+) T cells in Itk null mice have a phenotype of CD44(hi) similar to memory-like innate T cells. These cells are observed in mice carrying an Itk mutant lacking the kinase domain, indicating that active Tec kinase signaling suppresses their presence. These cells carry preformed message for and are able to rapidly produce IFN-gamma upon stimulation in vitro with IL-12/IL-18, and endow Itk null mice the ability to effectively respond to infection with L. monocytogenes or exposure to lipopolysaccharides by secretion of IFN-gamma. Transfer of these cells rescues the ability of IFN-gamma null mice to reduce bacterial burden following L. monocytogenes infection, indicating that these cells are functional CD8(+)CD44(hi) T cells previously detected in vivo. These results indicate that active signals from Tec kinases regulate the development of memory-like CD8(+) T cells with innate function.


Prognostic factors of Guillain-Barré syndrome: a 111-case retrospective review.
To identify the predictive factors associated with worse prognosis in the Guillain-Barré syndrome (GBS), which can be helpful to fully evaluate the disease progression and provide proper treatments. Clinical data of 111 GBS patients who were diagnosed from 2010 to 2015 were collected and retrospectively analyzed. Patients with diabetes (P=0.031), high blood pressure at admission (P=0.034), uroschesis (P=0.028), fever (P<0.001), ventilator support (P<0.001) during hospitalization, disorder of consciousness (p=0.007) and absence of preceding respiratory infection(P=0.016) were associated with worse outcome at discharge, while abnormal sensation, ataxia, weakness and decrease of tendon reflex seemed not correlated with the Medical Research Council(MRC) score at discharge. Compared with the subtype of acute inflammatory demyelinating polyneuropathy, prognosis of Miller-Fisher syndrome (p<0.001) and cranial nerve variant (p<0.038) were better, but prognosis of acute motor axonal neuropathy(AMAN) was worse (p<0.032). Laboratory examinations at admission showed that hyperglycemia (P=0.002), high leukocyte count (P=0.010), hyperfibrinogenemia (P=0.001), hyponatremia (P=0.020), hypoalbuminemia (P=0.005), abnormal hepatic (P=0.048) and renal (P=0.009) functions were associated with poorer prognosis at discharge, while albuminocytologic dissociation in cerebrospinal fluid, GM1 and GQ1b antibody showed no correlation with the MRC score at discharge. γ-Globulin and glucocorticoid therapies showed no difference in the MRC score at the discharge. AMAN, diabetes, high blood pressure, uroschesis, high body temperature, ventilator support, consciousness disorder, absence of upper respiratory tract preceding infection, hyperglycemia, hyponatremia, hypoalbuminemia, high leukocyte count, hyperfibrinogenemia, abnormal hepatic and renal function were demonstrated as poor prognostic factors.


Cartilage transformation in a composite graft of demineralized bovine bone matrix and ear perichondrium used in a child for the reconstruction of the nasal septum.
For the first time it was observed in a boy that a xenogenic implant of demineralized bovine bone matrix, enfolded in a pedicled flap of autogenic ear perichondrium, was transformed into autogenic cartilage. This new cartilage could be adequately used as a graft to bridge the cartilaginous defect of a nasal septum perforation. The defect was successfully closed by a technique employing four bipedicled mucosal advancement flaps of the septum and the nasal floor. The result of this pilot study, which is based on experiments in growing rabbits, appears to open the possibility to create de novo young and growing autogenic cartilage of substantial quantity in children, without the loss of the cartilage present.


Impact of constitutional polymorphisms in VCAM1 and CD44 on CD34+ cell collection yield after administration of granulocyte colony-stimulating factor to healthy donors.
Background The number of CD34(+) cells mobilized from bone marrow to peripheral blood after administration of granulocyte colony-stimulating factor varies greatly among healthy donors. This fact might be explained, at least in part, by constitutional differences in genes involved in the interactions tethering CD34(+) cells to the bone marrow. We analyzed genetic characteristics associated with CD34(+) cell mobilization in 112 healthy individuals receiving granulocyte colony-stimulating factor (filgrastim; 10 μg/kg; 5 days). Genetic variants in VCAM1 and in CD44 were associated with the number of CD34(+) cells in peripheral blood after granulocyte colony-stimulating factor administration (P = 0.02 and P = 0.04, respectively), with the quantity of CD34(+) cells ×10⁶/kg of donor (4.6 versus 6.3; P < 0.001 and 7 versus 5.6; P = 0.025, respectively), and with total CD34(+) cells ×10⁶ (355 versus 495; P = 0.002 and 522 versus 422; P = 0.012, respectively) in the first apheresis. Of note, granulocyte colony-stimulating factor administration was associated with complete disappearance of VCAM1 mRNA expression in peripheral blood. Moreover, genetic variants in granulocyte colony-stimulating factor receptor (CSF3R) and in CXCL12 were associated with a lower and higher number of granulocyte colony-stimulating factor-mobilized CD34(+) cells/μL in peripheral blood (81 versus 106; P = 0.002 and 165 versus 98; P=0.02, respectively) and a genetic variant in CXCR4 was associated with a lower quantity of CD34(+) cells ×10⁶/kg of donor and total CD34(+) cells ×10⁶ (5.3 versus 6.7; P = 0.02 and 399 versus 533; P = 0.01, respectively). Conclusions In conclusion, genetic variability in molecules involved in migration and homing of CD34(+) cells influences the degree of mobilization of these cells.


Intraorbital findings in giant cell arteritis on black blood MRI.
Blindness is a feared complication of giant cell arteritis (GCA). However, the spectrum of pathologic orbital imaging findings on magnetic resonance imaging (MRI) in GCA is not well understood. In this study, we assess inflammatory changes of intraorbital structures on black blood MRI (BB-MRI) in patients with GCA compared to age-matched controls. In this multicenter case-control study, 106 subjects underwent BB-MRI. Fifty-six patients with clinically or histologically diagnosed GCA and 50 age-matched controls without clinical or laboratory evidence of vasculitis were included. All individuals were imaged on a 3-T MR scanner with a post-contrast compressed-sensing (CS) T1-weighted sampling perfection with application-optimized contrasts using different flip angle evolution (SPACE) BB-MRI sequence. Imaging results were correlated with available clinical symptoms. Eighteen of 56 GCA patients (32%) showed inflammatory changes of at least one of the intraorbital structures. The most common finding was enhancement of at least one of the optic nerve sheaths (N = 13, 72%). Vessel wall enhancement of the ophthalmic artery was unilateral in 8 and bilateral in 3 patients. Enhancement of the optic nerve was observed in one patient. There was no significant correlation between imaging features of inflammation and clinically reported orbital symptoms (p = 0.10). None of the age-matched control patients showed any inflammatory changes of intraorbital structures. BB-MRI revealed inflammatory findings in the orbits in up to 32% of patients with GCA. Optic nerve sheath enhancement was the most common intraorbital inflammatory change on BB-MRI. MRI findings were independent of clinically reported orbital symptoms. • Up to 32% of GCA patients shows signs of inflammation of intraorbital structures on BB-MRI. • Enhancement of the optic nerve sheath is the most common intraorbital finding in GCA patients on BB-MRI. • Features of inflammation of intraorbital structures are independent of clinically reported symptoms.


The cerebellum: its role in language and related cognitive and affective functions.
The traditional view on the cerebellum as the sole coordinator of motor function has been substantially redefined during the past decades. Neuroanatomical, neuroimaging and clinical studies have extended the role of the cerebellum to the modulation of cognitive and affective processing. Neuroanatomical studies have demonstrated cerebellar connectivity with the supratentorial association areas involved in higher cognitive and affective functioning, while functional neuroimaging and clinical studies have provided evidence of cerebellar involvement in a variety of cognitive and affective tasks. This paper reviews the recently acknowledged role of the cerebellum in linguistic and related cognitive and behavioral-affective functions. In addition, typical cerebellar syndromes such as the cerebellar cognitive affective syndrome (CCAS) and the posterior fossa syndrome (PFS) will be briefly discussed and the current hypotheses dealing with the presumed neurobiological mechanisms underlying the linguistic, cognitive and affective modulatory role of the cerebellum will be reviewed.


Recombinant virus-like particles of a norovirus (genogroup II strain) administered intranasally and orally with mucosal adjuvants LT and LT(R192G) in BALB/c mice induce specific humoral and cellular Th1/Th2-like immune responses.
We investigated the immune response induced by mucosal immunization of BALB/c mice with virus-like particles (VLPs) of a genogroup II norovirus, Dijon171/96 virus, produced in the baculovirus system. VLPs administered alone by the intranasal route induced a high serum antibody response as well as fecal IgA, which were enhanced when the heat-labile Escherichia coli toxin or its non toxic mutant LT(R192G) was coadministered. In these conditions, the oral route was also efficient. Cytokine production by cells from different lymphoid tissues was then assessed after in vitro restimulation. A Th1/Th2-like response was observed in cervical lymph node and Peyer's patch (PP) cell cultures from mice intranasally or orally immunized with either adjuvant indicating that, on the assumption that T cells are the primary cells producing the cytokines after in vitro restimulation, specific T lymphocytes are present in the intestine after intranasal immunization.


Efficacy of antidepressants in child and adolescent depression: a meta-analytic study.
To examine whether antidepressant drugs are superior to placebo in the treatment of juvenile depression. Extensive literature search was done to retrieve all randomised controlled and all uncontrolled trials describing children and adolescents with a diagnosis of depression who underwent any antidepressant drug treatment. In order to combine results, separate analyses using random effect models were conducted first for controlled and then for both controlled and open studies. 18 controlled and 23 open trials were submitted to meta-analysis. Tricyclics showed no significant benefit over placebo. Odds ratios for SSRIs were 1.84 (95% CI 1.35-2.50) for controlled and 1.83 (95% CI 1.40-2.40) for controlled and uncontrolled studies suggesting a significant benefit over placebo. Combining all antidepressants also gave confidence interval excluding the value one. Despite some promising data concerning the use of SSRIs in the treatment of adolescent depression, caution is warranted until the long-term safety of these agents can be demonstrated. Insufficient data are available to judge even the short term merits of these agents in prepubertal children. There is no evidence to support the use of tricyclics in this population.


'Metamorphosis': computer-assisted rotational analysis and interspatial distribution of nuclear structures from serial ultrathin sections.
'Metamorphosis' is a group of computer programs designed to permit rapid three-dimensional analysis and generation of models of nuclei cells from digitized data. Infinite exterior views are possible using the rotation program and the user has the added ability to enter the object and peer out. This permits analysis of structural associations from any perspective. Since many chromosome associations cannot be abstracted without a computer, using rotation and cluster analysis reveals obscure relationships. This concept becomes important when proposing theories involving interactions between two or more objects. The program is written in BASICA and designed for use on IBM-compatible personal computers equipped with a digitizer. Three-dimensional transformations are accomplished via matrix manipulation which permits rapid generation of new images.


The provision of nutritional advice and care for cancer patients: a UK national survey of healthcare professionals.
People living with and beyond cancer often experience nutrition-related issues and should receive appropriate advice on nutrition that is consistent and evidence based. The aim of this study was to investigate current practice for the provision of nutritional care by healthcare professionals (HCPs) from a UK national survey produced by the National Institute for Health Research (NIHR) Cancer and Nutrition Collaboration. An online survey sent to professional groups and networks included questions on discussing nutrition, providing information, awareness of guidelines, confidence in providing nutritional advice, training and strategies for improving nutritional management. There were 610 HCPs who responded including nurses (31%), dietitians (25%), doctors (31%) and speech and language therapists (9%). The majority of HCPs discusses nutrition (94%) and provides information on nutrition (77%). However, only 39% of HCPs reported being aware of nutritional guidelines, and just 20% were completely confident in providing nutritional advice. Awareness of guidelines varied between the different professional groups with most but not all dietitians reporting the greatest awareness of guidelines and GPs the least (p = 0.001). Those HCPs with a greater awareness of guidelines had received training (p = 0.001) and were more likely to report complete confidence in providing nutritional advice (p = 0.001). Whilst HCPs discuss nutrition with cancer patients and may provide information, many lack an awareness of guidelines and confidence in providing nutritional advice. To ensure consistency of practice and improvements in patient care, there is scope for enhancing the provision of appropriate nutrition education and training.


Oleanolic acid.
Oleanolic acid (3β-hydroxyolean-12-en-28-oic acid) is a pentacyclic triterpenoid compound with a widespread occurrence throughout the plant kingdom. In nature, the compound exists either as a free acid or as an aglycone precursor for triterpenoid saponins, in which it can be linked to one or more sugar chains. Oleanolic acid and its derivatives possess several promising pharmacological activities, such as hepatoprotective effects, and anti-inflammatory, antioxidant, or anticancer activities. With the recent elucidation of its biosynthesis and the imminent commercialization of the first oleanolic acid-derived drug, the compound promises to remain important for various studies. In this review, the recent progress in understanding the oleanolic acid biosynthesis and its pharmacology are discussed. Furthermore, the importance and potential application of synthetic oleanolic acid derivatives are highlighted, and research perspectives on oleanolic acid are given.


Transfontanellar duplex brain ultrasonography resistive indices as a prognostic tool in neonatal hypoxic-ischemic encephalopathy before and after treatment with therapeutic hypothermia.
Prior to therapeutic hypothermia (that is, cooling), transfontanellar duplex brain sonography resistive indices (RI) were studied as a bedside non-invasive measures of cerebral hemodynamics in neonates who suffered from hypoxic-ischemic encephalopathy (HIE). We compared pre- and post-cooling RI values and examined the relationships between RI values and specific long-term neurodevelopmental outcomes. Transfontanellar duplex brain sonography, including RI, were obtained for 28 neonates prior to cooling and for 20 neonates following cooling. All RI values were sampled in the anterior cerebral artery at the beginning of each ultrasound study. Neurodevelopmental assessment was conducted between ages 20-32 months with the Mullen Scale of Early Learning. The relationships between pre- and post-cooling RI and cognitive and motor outcomes were studied. Neonates with RI values <0.60 prior to and following cooling were more likely to die or have severe neurodevelopmental disability by ages 20-32 months than those with RI>0.60. Lower RI values were associated with specific neurodevelopmental deficits in motor skill attainment. Pre- and post-cooling transfontanellar duplex brain sonography RI values may be a useful prognostic tool, in conjunction with other clinical information, for neonates diagnosed with HIE. The results of this study suggest that further study of the prognostic value of RI values for short- and long-term outcomes is warranted.


Experimental intraocular lens implantation in the rabbit eye and in the mouse peritoneal space. Part I: Cellular components observed on the implanted lens surface.
Polymethylmethacrylate and silicone intraocular lenses (IOLs) were implanted in the rabbit eye and in the mouse peritoneal space to study cellular response to the implanted IOL. Animals were sacrificed at various times after implantation. The removed IOLs were properly fixed and stained using Wolter's implant cytology technique. Scanning and transmission electron microscopic studies were performed. Macrophages, lymphocytes, and fibroblast-like cells were the major components identified. Occasional formations of foreign-body giant cells were seen. The responses observed on IOLs implanted in the rabbit eye were generally milder than those observed on IOLs implanted in the mouse peritoneal space.


Structural and computational analysis of peptide recognition mechanism of class-C type penicillin binding protein, alkaline D-peptidase from Bacillus cereus DF4-B.
Alkaline D-peptidase from Bacillus cereus DF4-B, called ADP, is a D-stereospecific endopeptidase reacting with oligopeptides containing D-phenylalanine (D-Phe) at N-terminal penultimate residue. ADP has attracted increasing attention because it is useful as a catalyst for synthesis of D-Phe oligopeptides or, with the help of substrate mimetics, L-amino acid peptides and proteins. Structure and functional analysis of ADP is expected to elucidate molecular mechanism of ADP. In this study, the crystal structure of ADP (apo) form was determined at 2.1 Å resolution. The fold of ADP is similar to that of the class C penicillin-binding proteins of type-AmpH. Docking simulations and fragment molecular orbital analyses of two peptides, (D-Phe)4 and (D-Phe)2-(L-Phe)2, with the putative substrate binding sites of ADP indicated that the P1 residue of the peptide interacts with hydrophobic residues at the S1 site of ADP. Furthermore, molecular dynamics simulation of ADP for 50 nsec suggested that the ADP forms large cavity at the active site. Formation of the cavity suggested that the ADP has open state in the solution. For the ADP, having the open state is convenient to bind the peptides having bulky side chain, such as (D-Phe)4. Taken together, we predicted peptide recognition mechanism of ADP.


Parkin Maintains Robust Pacemaking in Human Induced Pluripotent Stem Cell-Derived A9 Dopaminergic Neurons.
The degeneration of nigral (A9) dopaminergic (DA) neurons results in cardinal motor symptoms that define Parkinson's disease (PD). Loss-of-function mutations in parkin are linked to a rare form of early-onset PD that is inherited recessively. We generated isogenic human A9 DA neurons with or without parkin mutations to establish the causal relationship between parkin mutations and the dysfunction of human A9 DA neurons. Using TALEN (transcription activator-like effector nuclease)- or CRISPR/Cas9-mediated gene targeting, we produced two isogenic pairs of naivetropic induced pluripotent stem cells (iPSCs) by repairing exon 3 deletions of parkin in iPSCs derived from a PD patient and by introducing the PD-linked A82E mutation into iPSCs from a healthy subject. The four lines of isogenic iPSCs were differentiated to A9 DA neurons, which fired spontaneous pacemaking action potentials (AP) dependent on L-type Ca2+ channels. The frequency of the pacemaking APs was significantly reduced by parkin mutations introduced to normal neurons. Consistent with this, isogenic repair of parkin mutations significantly increased the frequency from that observed in patient-derived neurons. The results show that parkin maintains robust pacemaking in human iPSC-derived A9 DA neurons. The function is critical to normal DA transmission required for controlling voluntary locomotor activities. © 2023 International Parkinson and Movement Disorder Society.


Sudden Aggravated Radicular Pain Caused by Hemorrhagic Spinal Angiolipomas After Back Massage.
Spinal angiolipomas (SALs) are benign tumors that usually present a slow progressive spinal cord or radicular compression. Acute myelopathy or acute aggravated radicular syndrome are exceedingly rare. The authors report an original case with sudden aggravated radicular pain caused by hemorrhagic SALs. A 54-year-old woman presented with a 2-month history of mild back pain, and the pain was significantly aggravated after a therapeutic back massage. Neurologic examination showed pain and hyperalgesia between T4 and T7 dermatome, from back to front, just like a band. Spinal magnetic resonance imaging (MRI) showed a dorsally located epidural lesion (T4-T6) and a small intratumoral hemorrhage at the lower part of the tumor. A bilateral T4-T6 laminectomy was performed to achieve total excision of the tumors. Histologic examination showed that the tumors were composed of mature adipose tissue and vascular tissue as angiolipomas. The postoperative course was uneventful with complete neurologic recovery 4 days after the surgery. MRI at 1-year follow-up indicated no recurrence. SALs are unusual benign tumors that are composed of mature fatty tissue and abnormal blood vessels; sudden aggravated spinal cord or radicular compression syndrome is rare. MRI is the best choice in the diagnosis of SALs. Surgery may be performed in different ways depending on the type of SALs, and the prognosis is generally good.


CT spectral imaging for monitoring the therapeutic efficacy of VEGF receptor kinase inhibitor AG-013736 in rabbit VX2 liver tumours.
The aim of this study was to evaluate the value of computed tomography (CT) spectral imaging in assessing the therapeutic efficacy of a vascular endothelial growth factor (VEGF) receptor inhibitor AG-013736 in rabbit VX2 liver tumours. Twenty-three VX2 liver tumour-bearing rabbits were scanned with CT in spectral imaging mode during the arterial phase (AP) and portal phase (PP). The iodine concentrations(ICs)of tumours normalized to aorta (nICs) at different time points (baseline, 2, 4, 7, 10, and 14 days after treatment) were compared within the treated group (n = 17) as well as between the control (n = 6) and treated groups. Correlations between the tumour size, necrotic fraction (NF), microvessel density (MVD), and nICs were analysed. The change of nICs relative to baseline in the treated group was lower compared to the control group. A greater decrease in the nIC of a tumour at 2 days was positively correlated with a smaller increase in tumour size at 14 days (P < 0.05 for both). The tumour nIC values in AP and PP had correlations with MVD (r = 0.71 and 0.52) and NF (r = -0.54 and -0.51) (P < 0.05 for all). CT spectral imaging allows for the evaluation and early prediction of tumour response to AG-013736. • AG-013736 treatment response was evaluated by CT in a rabbit tumour model. • CT spectral imaging allows for the early treatment monitoring of targeted anti-tumour therapies. • Spectral CT findings correlated with vascular changes after anti-tumour therapies. • Spectral CT is a promising method for assessing clinical treatment response.


[Imaging of the temporomandibular joint].
The recent advances in imaging of the temporomandibular joint (TMJ), especially the introduction of MR imaging and surface colls allowing precise visualization of superficial structures led us to reconsider the different techniques used in this very complex anatomical region, due to both its morphology and function. We also tried to determine their respective role, especially in the study of TMJ dysfunction syndromes, that represent the most frequent pathology of this region. Conventional radiography allows us to appreciate the overall amplitude of the joint movements, and to study bone abnormalities, but CT is much more precise in the study of cortical bone. In TMJ dysfunction, the joint itself is studied by either arthrography or MRI, but both techniques have their limitations and remain complementary in some aspects, which are detailed here, so that the choice between them depends on availability and therapeutic indications.


Interaction of diazepam with synaptic transmission in the in vitro rat hippocampus.
Diazepam (5 x 10(-8) - 10(-6) M) was found to augment recurrent inhibition of pyramidal neuron firing in a dose-dependent manner in rat hippocampal slices. To determine possible loci of this effect, diazepam was locally administered by pressure ejection from a micropipette, while recording action potentials from single inhibitory ("basket") interneurons. Diazepam induced reversible and reliable increases in interneuron firing in response to stimulation of Schaffer collateral and commissural afferents. Taken together with previous electrophysiological reports, these data suggest that benzodiazepines may augment central inhibition by increasing either the excitability of inhibitory interneurons, or by increasing the strength of excitatory afferents to these cells.


Radiographic and ultrasonographic diagnosis of liver diseases in dogs and cats.
Various techniques utilized for the clinical investigation of small animal patients with signs of liver disease are discussed. The author reviews the evaluation of survey radiographs and selected radiographic contrast techniques, such as cholecystography. The author also outlines the basic principles of hepatic ultrasonography and includes examples of normal and abnormal hepatic ultrasonograms to demonstrate the application of this new imaging modality to the diagnosis of liver diseases.


Studies on fibrous tissues of the basilar membrane in inner ear.
Of a number of electron microscope studies on the basilar membrane of the inner ear cochlear duct so far reported, none seems to have been conducted using PAM of R.R. staining, excellent stainings especially suited for the observation of connective tissue. In this study the basilar membrane of the cochlear duct in young and mature guinea pigs and human fetuses was examined with an electron microscope using these stainings, and obtained the interesting results described below. Fibrils of the basilar membrane, although strongly PAM-positive, all lacked periodic stripes, indicating that they were collagenous fibrils in an immature state. Young guinea pigs clearly differed from mature ones in that numerous fibroblasts were present. In human fetuses, the basilar membrane exhibited no clear fibrillar structure in the 13th week, but had PAM-positive fibrils in the 20th week. By the 23rd week, it had assumed roughly the same structure as in adults, although numerous fibroblasts were still present, and resembled the basilar membrane in young guinea pigs. The basilar membrane is a part of a unique organ, a sensory organ. Though comparison is difficult because of the extremely complicated anatomy of the organ, it is concluded that the fibrils of basilar membrane are collagenous fibrils in a peculiarly immature state, and similar to those in the cartilage and tympanic membrane.


The effect of aging on cutaneous lipid peroxide levels and superoxide dismutase activity in guinea pigs and patients with burns.
Cutaneous lipid peroxide levels and superoxide dismutase (SOD) activity in non-aged and aged guinea pigs were measured between 15 min and 7 days after experimental infliction of burns. Skin burns on non-aged and aged patients were also subjected to these assays. In non-aged guinea pig skin burns, lipid peroxide levels increased from 24 hr to the fourth day after the burn infliction, while SOD activity did not increase but showed a slight decrease 12 hr and 24 hr post-burn. On the other hand, while the aged group showed a more increase in skin lipid peroxide levels compared to that seen in non-aged mice, skin SOD activity began to decrease from 30 min post-burn, the maximum decrease being reached on the second day. The activity did not return to normal by the 7th day. In non-aged patients skin burns showed increases in both lipid peroxide levels and SOD activity, while in aged patients, though they showed a marked increase in lipid peroxide levels, SOD activity remained unchanged. The present study indicated that, although in our recent study, skin SOD activity of healthy elderly people was found to be comparable to that in non-aged individuals, the capacity for induction of SOD activity under oxygen stress differed with age in both guinea pig and human burn sufferers. Furthermore, this induction capacity seemed to vary from species to species.


Sonographically observed echogenic response during intraoperative radiofrequency ablation of cirrhotic livers: pathologic correlation.
We performed a study to determine the correlation between the diameter of the echogenic response observed with intraoperative sonography during radiofrequency ablation of the cirrhotic liver and the mean diameter of tissue necrosis. A total of 22 intraoperative radiofrequency ablations were created in 11 cirrhotic livers. The largest diameter of the sonographically observed echogenic response surrounding and perpendicular to the radiofrequency probe was measured. The subsequent zone of necrosis observed at pathology in the hepatectomy specimens after liver transplantation was measured in three planes and compared with the measured diameter of the echogenic response. During all except three ablations, a hyperechoic region was visualized surrounding the radiofrequency probe. The diameter of the echogenic response correlated significantly with the mean diameter of necrosis (correlation coefficient, 0.84). However, the echogenic response overestimated the minimal diameter of necrosis (mean difference, 0.8 +/- 0.4 cm) in 18 of 22 ablations and underestimated the maximum diameter of necrosis (mean difference, 0.9 +/- 0.8 cm) in 16 of 22 ablations. The diameter of the echogenic response observed with intraoperative sonography during radiofrequency ablation of the cirrhotic liver correlates closely with the mean diameter of the subsequent area of tissue necrosis. However, the solitary diameter of the echogenic response as measured in our study was often greater than the smallest diameter and less than the largest diameter of the area of tissue necrosis. Therefore, the echogenic response associated with radiofrequency ablation of the cirrhotic liver should be viewed only as a rough approximation of the area of induced tissue necrosis; the final assessment of the adequacy of ablation should be deferred to an alternative imaging technique.


Host Differential Sensitization toward Color/Lifetime-Tuned Lanthanide Coordination Polymers for Optical Multiplexing.
Optical multiplexing based on luminescent materials with tunable color/lifetime has potential applications in information storage and security. However, the available tunable luminescent materials reported so far still suffer from several drawbacks of low efficiency or poor stability, thus restraining their further applications. Herein, we demonstrate a strategy to develop efficient and stable lanthanide coordination polymers (LCPs) with tunable luminescence as a new option for optical multiplexing. Their multicolor emission from green to red and naked-eye-sensitive green emission with tunable lifetime (from ca. 300 to ca. 600 μs) can be controlled by host differential sensitization and energy transfer between lanthanide ions. The quantum efficiencies of developed samples range from around 20 % to 46 % and the luminescence intensity/lifetime appear quite stable in polar solvents up to ten weeks. Furthermore, with the aid of inkjet printing and concepts of luminescence lifetime imaging and time-gated imaging, we illustrate their promising applications of information storage and security in spatial and temporal domains.


Large-conductance calcium-activated potassium channels modulate vascular tone in experimental cirrhosis.
Large-conductance calcium-activated potassium (BK(Ca)) channels regulate vascular tone in different vascular systems. Moreover, activated hepatic stellate cells (HSC) contain BK(Ca) channels. The aim of this study was to evaluate the role of BK(Ca) channels in the regulation of vascular tone in control (CT) and carbon tetrachloride-cirrhotic (CH) rat livers. Changes in intrahepatic vascular resistance were assessed by evaluating the portal perfusion pressure (PP) response to methoxamine (Mtx) in the presence of Iberiotoxin (Ibtx; a BK(Ca) channel blocker), NS1619 (a BK(Ca) channel opener), Ibtx plus the nitric oxide (NO) synthase inhibitor, N(G)-nitro-L-arginine (L-NNA) or L-NNA alone. In addition, in CH livers, PP dose-response curves to the NO donor, S-nitroso-N-acetyl-D,L-penicillamine (SNAP), were performed after pre-incubation with Ibtx or its vehicle. BK(Ca) mRNA expression was assessed in liver homogenates, and BK(Ca) protein expression in HSC isolated from CT and CH livers. In CH livers, Ibtx significantly increased baseline PP and exacerbated the PP response to Mtx. Conversely, NS1619 induced a mild nonsignificant decrease of baseline PP and attenuated the hyperresponse to Mtx. CH livers exhibited an upregulation of both mRNA and protein of the alpha-subunit of BK(Ca). Large-conductance calcium-activated potassium channels are overexpressed in CH livers and might represent a compensatory mechanism modulating the increased hepatic vascular tone of cirrhosis.


Detection of astrovirus in the faeces of cats with diarrhoea.
Examination by electron microscopy of faeces from two separate cases of young cats with diarrhoea revealed the presence of 28 nm viral particles morphologically consistent with an astrovirus. No visible cytopathic effect was observed when the virus was inoculated into a feline kidney cell culture.


A look into stent-related thrombus-burden: Bivalirudin versus heparin.
Use of optical coherence tomography (OCT) adds an assessment of thrombus burden remaining on stents after PCI for acute coronary syndromes. Potential variations in stent-related thrombus burden can be documented by OCT as a function of peri-procedural pharmacology supporting the use of OCT in future hypothesis testing. Bivalirudin remains a reliable and expensive alternative to heparin in cases of HIT or patients at high bleeding risk during transfemoral PCI.


Complications in colonoscopic polypectomy: lessons to learn from an experience with 1576 polyps.
One thousand five hundred and seventy six polyps were removed from 1190 patients. Polyp size ranged from 5 mm to 6 cm. Nineteen complications occurred, an incidence of 1.2 per cent. Bleeding was the most common complication, accounting for 53 per cent of all complications, followed by transmural burn (32%). Other complications included a "silent" free perforation, a snare-wire entrapment, and an ensnared bowel wall. Certain complications can be prevented provided the basic safety features are observed. Some complications require conservative management, others require an operation.


Comprehensive study into the activation of the plasma enzyme systems during attacks of hereditary angioedema due to C1-inhibitor deficiency.
The activation of plasma enzyme systems contributes to hereditary angioedema attacks. We aimed to study the activation markers of the fibrinolytic, coagulation, and contact systems in a larger number of paired samples obtained from the same C1-INH-HAE patients in symptom-free periods and during attacks. Eleven parameters (Factors XI, XII, and C1-inhibitor activity; the concentrations of the D-dimer, prothrombin fragments 1 + 2, plasminogen, plasminogen activator inhibitor-1 [PAI-1], thrombin-anti-thrombin III [TAT] complex, fibrinogen) were measured along with prothrombin time and activated partial thromboplastin time (aPTT), using commercial kits. We compared these markers in samples obtained from the same 39 patients during attack-free periods and during 62 edematous episodes. Forty healthy subjects of matching sex and age served as controls. Compared with the healthy controls, significantly higher FXI and FXII activity (p = 0.0007, p = 0.005), as well as D-dimer (p < 0.0001), prothrombin fragments 1 + 2 (p < 0.0001), and TAT (p = 0.0303) levels were ascertained in the patients during symptom-free periods. The evaluation of samples from symptom-free periods or obtained during attacks revealed the increase of FXII activity, as well as of the concentration of D-dimer, prothrombin fragments 1 + 2, and TAT during edematous episodes. PAI-1 level, prothrombin time, and aPTT decreased significantly during attacks, compared with symptom-free periods. D-dimer level was significantly higher during multiple- vs. single-site attacks. Comparing a large number of paired samples from symptom-free periods or from edematous episodes allowed accurate appraisal of the changes occurring during attacks. Moreover, our study pointed out that individual episodes may be characterized by different marker patterns.


Phenylboronic-acid-based nanocomplex as a feasible delivery platform of immune checkpoint inhibitor for potent cancer immunotherapy.
Inspired by our previous study, we report a simple yet effective platform for therapeutic antibody delivery. A polymeric phenylboronic acid (pPBA)-antibody nanocomplex was simply formulated by mixing pPBA and antibody, derived by the formation of a pH-responsive phenylboronic ester between the PBA group on pPBA and diol on the inherent glycosylation site of the antibody. We focused on the basic prerequisites for a successful delivery, protection from degradation during the circulation, and release at the target lesion. To evaluate the antibody delivery system, anti-PD-L1, one of the most common antibody therapeutics in immuno-oncology, and mouse colon cancer model with an MC-38 cell line were used. Several in-vitro assays reveal the outstanding protective effect of the nanocomplexes as well as the pH-responsive release of antibodies. Moreover, the anti-PD-L1 nanocomplex exhibited an enhanced circulation as well as a better accumulation in tumor lesions after administration in vivo, which led to a significant antitumor effect in comparison to that of a free antibody. Our nanocomplex platform is a promising antibody delivery system for application in conventional antibody-mediated therapies.


Endocrine and protein response to elective surgery: effect of nursing at two different environmental temperatures.
Twenty male patients undergoing elective abdominal surgery were studied on one control day pre-operatively and for the first 4 days postoperatively at an environmental temperature of either 20 degrees C or 28 degrees C. Glucose, insulin, growth hormone and urinary catecholamine levels all rose significantly on the first postoperative day returning to within normal limits by day 4. Urine nitrogen excretion also increased significantly postoperatively in both groups. Environmental temperature did not influence these responses. Concentrations of C-reactive protein and a1-antichymotrypsin increased significantly on the first postoperative day and were returning to normal by day 4 while concentrations of haptoglobin, a1-antitrypsin and a1-acid glycoprotein, which also showed a significant increase, remained elevated at the end of the study. Concentrations of albumin, transferrin, a2-macroglobulin, prealbumin and retinol binding protein all decreased significantly following operation. Elevated environmental temperature did not affect the protein response to surgery. Urine cortisol and 17-hydroxycorticosteroid excretion rose significantly postoperatively and remained elevated throughout the study. Patients nursed at 28 degrees C showed significantly lower postoperative excretion of both hormones than patients nursed at 20 degrees C.


Effect of Simvastatin Treatment on Mitochondrial Function and Inflammatory Status of Human White Adipose Tissue.
Statin therapy has shown pleiotropic effects affecting both mitochondrial function and inflammatory status. However, few studies have investigated the concurrent effects of statin exposure on mitochondrial function and inflammatory status in human subcutaneous white adipose tissue. In a cross-sectional study, we investigated the effects of simvastatin on mitochondrial function and inflammatory status in subcutaneous white adipose tissue of 55 human participants: 38 patients (19 females/19 males) in primary prevention with simvastatin (> 40 mg/d, > 3 mo) and 17 controls (9 females/8 males) with elevated plasma cholesterol. The 2 groups were matched on age, body mass index, and maximal oxygen consumption. Anthropometrics and fasting biochemical characteristics were measured. Mitochondrial respiratory capacity was assessed in white adipose tissue by high-resolution respirometry. Subcutaneous white adipose tissue expression of the inflammatory markers IL-6, chemokine (C-C motif) ligand 2 (CCL2), CCL-5, tumor necrosis factor-α, IL-10, and IL-4 was analyzed by quantitative PCR. Simvastatin-treated patients showed lower plasma cholesterol (P < .0001), low-density lipoprotein (P < .0001), and triglyceride levels (P = .0116) than controls. Simvastatin-treated patients had a lower oxidative phosphorylation capacity of mitochondrial complex II (P = .0001 when normalized to wet weight, P < .0001 when normalized to citrate synthase activity [intrinsic]), and a lower intrinsic mitochondrial electron transport system capacity (P = .0004). Simvastatin-treated patients showed higher IL-6 expression than controls (P = .0202). Simvastatin treatment was linked to mitochondrial respiratory capacity in human subcutaneous white adipose tissue, but no clear link was found between statin exposure, respiratory changes, and inflammatory status of adipose tissue.


Localization and characterization of prolactin-like immunoreactivity in the pituitary of the frog Rana ridibunda.
Distribution and quantification of PRL in the pituitary gland of the frog Rana ridibunda were investigated using a high-affinity antiserum raised against bullfrog prolactin (PRL). The immunoreactive PRL-producing cells were distributed throughout the pars distalis, the highest density of cells being observed in the rostral region of the adenohypophysis facing the neurointermediate lobe. The dorsal region of the pars distalis contained only a few scattered PRL-immunoreactive cells. At the electron microscopic level, PRL-containing cells were visualized using the immunogold procedure. PRL-immunoreactive material was exclusively stored in secretory granules (size ranging from 200 to 700 nm in diameter). Neither the rough endoplasmic reticulum nor the Golgi apparatus were immunolabeled. Using a radioimmunoassay method we have compared the displacement curves obtained with bullfrog PRL and acetic extracts from Rana ridibunda pituitary. The two binding curves were not completely parallel, suggesting the existence of slight variations of the amino acid sequences of PRL in the two species. The concentration of PRL in the green frog adenohypophysis appeared somewhat higher (35.3 +/- 8.8 micrograms/mg protein) than that in the bullfrog pituitary. These results validate the use of an antiserum to bullfrog PRL to investigate the regulation of PRL secretion in Rana ridibunda.


Effect of the state of oxidation of cysteine 190 of tropomyosin on the assembly of the actin-tropomyosin complex.
Tropomyosin, cross-linked at cysteine 190, was found to bind more weakly to actin filaments than uncross-linked tropomyosin. Cross-linking of tropomyosin can cause actin filaments nearly completely covered with tropomyosin to be uncovered almost completely. The critical monomer concentration of actin is not significantly changed by binding of cross-linked or uncross-linked tropomyosin to actin filaments. The binding curves were analyzed quantitatively, thereby taking into account the polar end-to-end contact of tropomyosin molecules bound by actin and the overlap of the seven subunit binding sites along the actin filament. Under the conditions of the experiment (80 mM KCl, 1 mM MgCl2, pH 7.5, 38-42 degrees C), the equilibrium constant for isolated binding of tropomyosin to actin filaments is in the range 1 x 10(3)-3 x 10(3) M-1. The equilibrium constants for binding of tropomyosin to binding sites along the actin filament with one or two neighbouring tropomyosin molecules are in the range of 10(6) or 10(8) to 10(9) M-1, respectively. The equilibrium constants for binding of tropomyosin to binding sites along the actin filament with one or two neighbouring tropomyosin molecules are in the range of 10(6) or 10(8) to 10(9) M-1, respectively. The equilibrium constants for cross-linked and uncross-linked tropomyosin differ by a factor of only about two. Owing to the highly cooperative binding, these differences are sufficient so that actin filaments nearly completely covered with uncross-linked tropomyosin are uncovered almost completely by cross-linking tropomyosin at cysteine 190.


Protocatechuic acid induces cell death in HepG2 hepatocellular carcinoma cells through a c-Jun N-terminal kinase-dependent mechanism.
Protocatechuic acid (PCA), chlorogenic acid (CA) and luteolin (LT) are plant phenols found in Chinese medicinal herbs such as Lonicera japonica. Cytotoxicity assays showed that PCA, CA and LT (at 100 micromol/L) effectively killed the HepG2 hepatocellular carcinoma cells. Among these three naturally occurring compounds, only PCA was capable of stimulating the c-Jun N-terminal kinase (JNK) and p38 subgroups of the mitogen-activated protein kinase (MAPK) family. Coincidently, PCA-induced cell death was rescued by specific inhibitors for JNK and p38, while the cytotoxicities of CA and LT were partially eliminated by the antioxidant effect of N-acetyl-L-cysteine (NAC). Further investigation demonstrated that the aqueous extract of Lonicera japonica also triggered HepG2 cell death in a JNK-dependent manner, but the amount of PCA alone in this herbal extract was insufficient to contribute the subsequent cytotoxic effect. Collectively, our results suggest that PCA is a naturally occurring compound capable of inducing JNK-dependent hepatocellular carcinoma cell death.


Acid dye removal: comparison of surfactant-modified mesoporous FSM-16 with activated carbon derived from rice husk.
Cetyltrimethylammonium bromide (CTAB)-modified mesoporous molecular sieve FSM-16, prepared by a hydrothermal process (373 K, 3 days), was tested as an adsorbent for acid dye (acid yellow, AY, and acid blue, AB) removal in comparison with as-prepared FSM-16 and activated carbon (AC) derived from rice husk (50 vol% H3PO4, 773 K, 2.5 h). The adsorption isotherms, sorption kinetics, and pH effects upon acid dyes sorption on the adsorbents were thoroughly investigated. The structures of different adsorbents were characterized by XRD, FTIR spectroscopy, N2 adsorption measurements, and thermogravimetric (TG) analysis. It was found that the ultimate capacity of the adsorbents varied in the order FSM-16 > modified FSM-16 > AC and followed first-order rate kinetics. The adsorption isotherm of acid dyes on FSM-16 is of type IV, according to the IUPAC classification, drastically different from that of CTAB/FSM-16, which showed a type I isotherm. The latter sample had better adsorption performance at low concentration of acid dyes than the former. As compared to activated carbon of microporous character, the CTAB/FSM-16 sample achieved higher performance at low concentrations. This was due to the successful narrowing of the pore opening of FSM-16 using CTAB with maintenance of a considerable portion of the pore volume. Powder XRD and N2 adsorption studies of the CTAB/FSM-16 material indicated that the textural properties of the support were preserved during the hydrothermal synthesis and that the channels remained accessible, despite a significant reduction in surface area (ca. 26%). TG studies, on the other hand, confirmed that the modified material presented a higher hydrophobicity than that of the CTAB-free FSM-16 sample.


Concentrates of DHA from fish oil by selective esterification of cholesterol by immobilized isoforms of lipase from Candida rugosa.
Two lipases (Lip A and Lip B), were purified from a commercial lipase preparation produced by Candida rugosa and partially characterized. The purified lipases were immobilized on Duolite A 568 and used in the selective esterification of cholesterol with free fatty acids from sardine fish oil. The results showed that Lip A and Lip B preferentially esterified saturated and monounsaturated fatty acids allowing a 3.4-fold (Lip B, 24 h) and 4-fold (Lip A, 10 h) enrichment of docosahexaenoic acid in the remaining free fatty acid fraction. Selectivity towards eicosapentaenoic acid was less pronounced. By this selective esterification docosahexaenoic acid was concentrated from 7.4 to 32% with a recovery of 95% of its initial content in sardine fish oil.


A comparison of simultaneous organic carbon and nitrogen removal in microbial fuel cells and microbial electrolysis cells.
This study demonstrates simultaneous carbon and nitrogen removal in laboratory-scale continuous flow microbial fuel cell (MFC) and microbial electrolysis cell (MEC) and provides side-by side comparison of these bioelectrochemical systems. The maximum organic carbon removal rates in MFC and MEC tests were similar at 5.1 g L(-1) d(-1) and 4.16 g L(-1) d(-1), respectively, with a near 100% carbon removal efficiency at an organic load of 3.3 g L(-1) d(-1). An ammonium removal efficiency of 30-55% with near-zero nitrite and nitrate concentrations was observed in the MFC operated at an optimal external resistance, while open-circuit MFC operation resulted in a reduced carbon and ammonium removal of 53% and 21%, respectively. In the MEC ammonium removal was limited to 7-12% under anaerobic conditions, while micro-aerobic conditions increased the removal efficiency to 31%. Also, at zero applied voltage both carbon and ammonium removal efficiencies were reduced to 42% and 4%, respectively. Based on the observed performance under different operating conditions, it was concluded that simultaneous carbon and nitrogen removal was facilitated by concurrent anaerobic and aerobic biotransformation pathways at the anode and cathode, which balanced bioelectrochemical nitrification and denitrification reactions.


Fusarium toxins in cereals - Results from eight German Federals States in 2000.
A total of 353 cereal samples from 8 German Federal States were analysed for Fusarium toxins in the 2000' survey. The level of mycotoxin contamination of the samples was higher than in the previous year due to higher moisture during flowering in some regions. Concerning deoxynivalenol (DON), the prevailing mycotoxin in all the samples, we have to consider, that 24% of the samples as well as the average concentration exceed the advisory level of 500 μg/kg. The application of Fusarium active fungicides reduced the DON level from 963 μg/kg (samples without treatment) to 630 μg/kg (all samples from treated and untreated fields). It has to be emphasised that the number of nivalenol (NIV) positive samples increased in comparison to the previous year from 9% to 17%. In addition, an increase in the level of NIV was detected, the maximum concentration in winter wheat was 3480 μg/kg. Cereals were rather moderately contaminated with zearalenone (ZEA), more than 80% of the samples contained less than 50 μg/kg.


Pediatric inflammatory bowel disease.
The treatment of children and families affected by pediatric IBD requires an awareness of the diverse psychobiologic effects of the disorder over the course of child, adolescent [53], and adult development [52]. Optimal treatment requires careful coordination of various medical, educational, and rehabilitative services and concerned, empathetic, continuity of care by knowledgeable clinicians [4]. In many cases, this care must also include individual psychotherapeutic work with affected youngsters at crucial junctures to help deal with the social and psychological stresses of IBD, as well supportive casework for parents and siblings.


Artificial intervertebral disc replacement using bioactive three-dimensional fabric: design, development, and preliminary animal study.
A new artificial intervertebral disc was developed, and its intrinsic biomechanical properties, bioactivity, and the effectiveness as a total disc replacement were evaluated in vitro and in vivo. To introduce a new artificial intervertebral disc and to evaluate the in vitro mechanical properties, fusion capacity to bone, and segmental biomechanics in the total intervertebral disc replacement using a sheep lumbar spine. The loss of biologic fusion at the bone-implant interface and prosthetic failures have been reported in previous artificial discs. There have been no clinically applicable discs with detailed experimental testing of in vivo mechanics and interface fusion capacity. The artificial intervertebral disc consists of a triaxial three-dimensional fabric (3-DF) woven with an ultra-high molecular weight polyethylene fiber, and spray-coated bioactive ceramics on the disc surface. The arrangement of weave properties was designed to produce mechanical behavior nearly equivalent to the natural intervertebral disc. Total intervertebral disc replacement at L2-L3 and L4-L5 was performed using 3-DF disc with or without internal fixation in a sheep lumbar spine model. The segmental biomechanics and interface histology were evaluated after surgery at 4 and 6 months. The tensile-compressive and torsional properties of prototype 3-DF were nearly equivalent to those of human lumbar disc. The lumbar segments replaced with 3-DF disc alone showed a significant decrease of flexion-extension range of motion to 28% of control values as well as partial bony fusion at 6 months. However, the use of temporary fixation provided a nearly physiologic mobility of the spinal segment after implant removal as well as excellent bone-disc fusion at 6 months. An artificial intervertebral disc using a three-dimensional fabric demonstrated excellent in vitro and in vivo performance in both biomechanics and interface histology. There is a potential for future clinical application.


Influence of protein free aqueous extract of parathyroid powder on serum vitamin A level in rats.
Administration of the protein-free extract of bovine parathyroid powder to rats resulted in a significant increase of the serum vitamin A level.


Purification and some properties of the nitrite reductase from the cyanobacterium Phormidium laminosum.
Assimilatory ferredoxin-nitrite reductase (EC 22.214.171.124, ammonia: ferredoxin oxidoreductase) has been purified 5300-fold with a specific activity of 625 units/mg protein from the filamentous non-heterocystous cyanobacterium Phormidium laminosum. The enzyme was soluble and consisted of a single polypeptidic chain of 54 kDa. It catalyzed the reduction of nitrite to ammonia using ferredoxin or flavodoxin as electron donor. Methyl and benzyl viologens were also effective as electron donors but neither flavins nor NAD(P)H were. The apparent Michaelis constants for nitrite, ferredoxin and methyl viologen were 40, 22 and 215 microM, respectively. Nitrite reductase activity was inhibited effectively by cyanide and thiol reagents. The enzyme exhibited absorption maxima at 281, 391 (Soret), 570 (alpha) and 695 nm, with epsilon 391 of 4.3 x 10(4) M-1 cm-1, and an absorbance ratio A281/A391 of 1.95, suggesting the presence of siroheme as prosthetic group. These results show that this enzyme is similar to those of eukaryotic organisms.


Solubilization and assay of phospholipase A2 activity from rat jejunal brush-border membranes.
The phospholipase activity of rat jejunal brush-border membranes was examined in the presence of several solubilizing agents, by measuring the hydrolysis of endogenous membrane phospholipids, as well as the hydrolysis of exogenous, radiolabelled substrates. Enzyme activity was highly stimulated by dispersion in 1% solutions of bile salts, or in a synthetic, bile-salt derivative, 3-[(3-cholamidopropyl)dimethylammonio]propanesulphonate (CHAPS). Under these conditions the endogenous membrane phospholipids were largely degraded to free fatty acids and water-soluble phosphate. In the presence of 1% CHAPS, hydrolysis of exogenous phosphatidylcholine was shown to be due to an initial phospholipase A2-type attack followed by a subsequent lysophospholipase-type attack. These activities co-purified with the brush-border membrane. Maximal phospholipase A2 hydrolysis occurred at an alkaline pH of 8-11, with bile-salt detergents present at greater than their critical micellar concentrations. Hydrolysis was completely divalent-ion independent. Phospholipase A2 activity was not stimulated by 50% diethyl ether or ethanol, or in the presence of 1% solutions of Triton X-100, Zwittergent 3-12, sodium dodecyl sulphate, or n-octylglucoside. Stimulation of phospholipase activity by detergents was not related to their effectiveness at solubilizing the membrane proteins. When assayed individually phosphatidylcholine and lysophosphatidylcholine were each hydrolyzed (at the sn-2 and sn-1 positions, respectively) at a rate of approximately 125 nmol/mg protein per min. When assayed together, the two substrates appeared to compete for the same active site over a wide range of concentrations. It was concluded that the brush-border membrane contains an integral membrane protein with phospholipase A2 and lysophospholipase activities, which is specifically stimulated by bile salts and bile salt-like detergents.