Isolation and characterization of cell wall-defective variants of Bacillus subtilis and Bacillus licheniformis.
Quantitative mass conversion of intact bacterial cells of Bacillus subtilis and B. licheniformis to L-phase variants has been effected after lysozyme treatment. After subculture of the unstable L-phase variants for several months in the presence of methicillin, stable L-phase variants were obtained which grew in the absence of the antibiotic and were then unable to revert to the classical bacterial phase under conditions which gave rise to mass reversion of the protoplasts and unstable L-variants. These stable L-phase variants, which retained many of the physiological properties of the bacterium from which they were derived, were capable of growing exponentially and multiplying in liquid medium. Their morphology and apparent modes of reproduction were consistent with that described for other L-phase variants. The morphological events, as monitored by the electron microscope, of the reversion to the intact bacterial phase of an unstable L-phase variant of B. licheniformis are described.


[Effect of hypothermia therapy on serum GFAP and UCH-L1 levels in neonates with hypoxic-ischemic encephalopathy].
To evaluate the effect of hypothermia therapy on serum glial fibrillary acidic protein (GFAP) and ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) levels in neonates with hypoxic-ischemic encephalopathy (HIE). Sixty-four HIE neonates were enrolled in this study. Thirty-three neonates with mild HIE were given conventional treatment and 31 neonates with moderate or severe HIE received conventional treatment and hypothermia therapy. Serum levels of GFAP and UCH-L1 were measured using ELISA before treatment and 6-12 hours after treatment. Serum levels of IL-6, IL-8, GFAP and UCH-L1 in the moderate/severe HIE group were significantly higher than in the mild HIE group (P<0.05) before treatment. Serum GFAP level was positively correlated with serum IL-6 (r=0.54; P<0.05) and IL-8 levels (r=0.63; P<0.05), while negatively correlated with Apgar score (r=-0.47, P<0.05). After treatment, serum levels of IL-6, IL-8 and UCH-L1 in the moderate/severe HIE group were significantly reduced (P<0.05), while serum GFAP levels increased significantly (P<0.05). The patients with abnormal neurological development showed higher serum GFAP levels than those with favourable prognosis (P<0.05). Receiver operating characteristic (ROC) curves analysis demonstrated that the area under curve (AUC) of GFAP and UCH-L1 were 0.714 and 0.703 respectively. At a cut-off value of 0.07 ng/mL, the sensitivity and specificity of GFAP for the diagnosis of HIE were 77% and 78% respectively. Hypothermia therapy can decrease serum UCH-L1 levels and increase serum GFAP levels in neonates with HIE. Based on their diagnostic value of brain injury, GFAP and UCH-L1 are promising to be novel biomarkers for HIE.


Cytotoxic Action of Palladium-Based Compound on Prostate Stem Cells, Primary Prostate Epithelial Cells, Prostate Epithelial Cells, and Prostate Cell Lines.
Prostate cancer is one of the most common types of cancer found to occur in males and is ranked as the second-highest cause of cancer-associated deaths among male patients. In this study, we have shown the influence of a new palladium-based anticancer agent in contrast to the six distinct prostate cancer lines and the primary cultures. In this study, we have used six distinct prostate cell lines, that is, PNT2-C2, LNCaP, BPH-1, PC-3, PNT1A, and P4E6. The MTP and ATP assay were performed to evaluate the growth of the cell and the flow cytometry to investigate the status of the cell cycle. The antigrowth effect of the palladium complex was evaluated against different cell lines at three time zones 24 h, 48 h, and 72 h. [PdCl(terpy)] (capsule)-2H2O is synthesized by direct encapsulation of equimolar amounts of capsule ions into [Pd (terpy) Cl] Cl-2H2O. A comparative analysis was done on 25 mM etoposide and 12 mM cisplatin, cytotoxic agents. The lowest IC50 value at 72 hours was 0.128 mM for BPH-1 cell lines with 0.139 mM, whereas PNT2-C2 cells were found to be most resistant with IC50 values of 0.829 mM. The antigrowth effect of palladium complex on cell lines was measured using the MTS assay at 24, 48, and 72 hours. BPH-1, PNT2-C2, and PNT1A either possess normal tissues or have benign prostatic hyperplasia tissues whereas P4E6, PC-3, and LNCaP cell lines possess malignant origin. The Pd complex exhibited significant cytotoxic action in stem cells when compared against etoposide. An antigrowth effect was reported for Pd complex at lower concentration, but it was more cytotoxic than etoposide with significant cytotoxicity (P=0.001). The palladium complex experienced a substantial antigrowth influence over most of the prostate tumor cell lines and the primary cultures, eventually, leading to the implementation of this Pd complex in the treating procedure of metastatic prostate cancer, which is tremendously resistant to the traditional treatment.


Updated guidelines on digital media use by children.
Given the speed at which mobile media has grown recently, the updated American Academy of Pediatrics guidelines for digital technology use by children are a welcome addition to the toolbox of pediatric healthcare providers. This article discusses the changes to the guidelines as well as other useful current research not specifically included in those guidelines.


Functionalized Molybdenum Disulfide Nanosheets for 0D-2D Hybrid Nanostructures: Photoinduced Charge Transfer and Enhanced Photoresponse.
The high-concentration stable dispersion of free-standing mono- or few-layer transition metal dichalcogenide (TMD) nanosheets (NSs) remains a significant barrier for their application in solution-processed optoelectronic devices. Here, we report oleylamine (OLA)- and dodecanethiol (DDT)-assisted exfoliation of MoS2 NSs in nonpolar organic solvent 1,2-dichlorobenzene (DCB), which enables high-concentration stable dispersion of free-standing mono- or few-layer NSs. The functionalized MoS2 NSs were further utilized for the fabrication of solution-processed 0D-2D hybrids of CuInS2 quantum dots (CIS QDs) and MoS2 NSs. The strong photoluminescence (PL) quenching and decreased PL lifetimes of CIS QDs attached to MoS2 NSs indicates efficient charge transfer from photoexcited CIS to MoS2 NSs. The photocurrent of CIS/MoS2 hybrid devices is dramatically enhanced compared to that of pure CIS and pristine MoS2-based devices, confirming that efficient charge separation and transfer occur from CIS QDs to MoS2 NSs.


Comparative effects of phthalate monoesters on gap junctional intercellular communication and peroxisome proliferation in rodent and primate hepatocytes.
Several phthalate esters, compounds used as plasticizers in a variety of commercial products, have been shown to induce hepatic tumors in rodents. In this study, the comparative effects of phthalate monoesters on inhibition of gap junctional intercellular communication and induction of peroxisomal beta-oxidation were assessed in primary cultured hepatocytes from rats, mice, hamsters, cynomolgus monkeys, and humans. A human liver cell line was also utilized. Eight monoesters examined included mono-2-ethylhexyl phthalate (MEHP), mono-n-octyl phthalate (MNOP), mono-isononyl phthalate (MINP, 3 types, -1, -2, and -3), mono-isoheptyl phthalate (MIHP), mono-isodecyl phthalate (MIDP), and mono-(heptyl, nonyl, undecyl) phthalate (M711P). Gap junctional intercellular communication was measured 4 and 24 h after treatment by lucifer yellow dye coupling. Gap junctional intercellular communication was inhibited in rat and mouse hepatocytes by all eight monoesters in a concentration-dependent manner. In most cases, gap junctional intercellular communication was significantly reduced at the lowest concentrations tested (50 pM). Inhibition of gap junctional intercellular communication in rodent cells was substantially reversed within 24 h of monoester removal. In contrast, cell-to-cell communication was not inhibited in hamster, cynomolgus, or human hepatocytes or in a human liver cell line at any concentration examined. In rat hepatocytes, peroxisomal beta-oxidation was elevated after treatment with MEHP, MINP, MIHP, and MIDP but not MNOP or M711P, and with all but MIHP in mouse hepatocytes. The eight phthalates produced no marked change on peroxisomal beta-oxidation in hepatocytes from other species. These data provide additional evidence that the toxicological effects of phthalate esters are species specific.


Spindle cell fragments in focal nodular hyperplasia of the liver. A case report.
There are only few reports on the fine needle aspiration cytology (FNAC) findings of focal nodular hyperplasia (FNH) of the liver. A 30-year-old woman who had undergone surgery for a leiomyosarcoma of the calf, was found to have a hepatic mass five years later on imaging during routine follow-up. Fine needle aspiration was performed to rule out metastasis. Cytology revealed a few fragments of bland-looking spindle cells in a metachromatic stroma along with benign hepatocytes and bile duct cells. It was interpreted as "consistent with metastasis of leiomyosarcoma." The excised mass showed histologic features typical of FNH. Spindle cell fragments have not been previously observed in the FNAC of FNH. These fragments probably represent the muscular wall of the abnormal blood vessels of FNH. If smooth muscle fragment is seen accompanying benign hepatocytes and bile duct cells, one should consider the diagnosis of FNH in the needle aspirate.


PaxDb 5.0: Curated Protein Quantification Data Suggests Adaptive Proteome Changes in Yeasts.
The "Protein Abundances Across Organisms" database (PaxDb) is an integrative metaresource dedicated to protein abundance levels, in tissue-specific or whole-organism proteomes. PaxDb focuses on computing best-estimate abundances for proteins in normal/healthy contexts and expresses abundance values for each protein in "parts per million" in relation to all other protein molecules in the cell. The uniform data reprocessing, quality scoring, and integrated orthology relations have made PaxDb one of the preferred tools for comparisons between individual datasets, tissues, or organisms. In describing the latest version 5.0 of PaxDb, we particularly emphasize the data integration from various types of raw data and how we expanded the number of organisms and tissue groups as well as the proteome coverage. The current collection of PaxDb includes 831 original datasets from 170 species, including 22 Archaea, 81 Bacteria, and 67 Eukaryota. Apart from detailing the data update, we also present a comparative analysis of the human proteome subset of PaxDb against the two most widely used human proteome data resources: Human Protein Atlas and Genotype-Tissue Expression. Lastly, through our protein abundance data, we reveal an evolutionary trend in the usage of sulfur-containing amino acids in the proteomes of Fungi.


Selection of appropriate reference genes for RT-qPCR analysis in a streptozotocin-induced Alzheimer's disease model of cynomolgus monkeys (Macaca fascicularis).
Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) has been widely used to quantify relative gene expression because of the specificity, sensitivity, and accuracy of this technique. In order to obtain reliable gene expression data from RT-qPCR experiments, it is important to utilize optimal reference genes for the normalization of target gene expression under varied experimental conditions. Previously, we developed and validated a novel icv-STZ cynomolgus monkey model for Alzheimer's disease (AD) research. However, in order to enhance the reliability of this disease model, appropriate reference genes must be selected to allow meaningful analysis of the gene expression levels in the icv-STZ cynomolgus monkey brain. In this study, we assessed the expression stability of 9 candidate reference genes in 2 matched-pair brain samples (5 regions) of control cynomolgus monkeys and those who had received intracerebroventricular injection of streptozotocin (icv-STZ). Three well-known analytical programs geNorm, NormFinder, and BestKeeper were used to choose the suitable reference genes from the total sample group, control group, and icv-STZ group. Combination analysis of the 3 different programs clearly indicated that the ideal reference genes are RPS19 and YWHAZ in the total sample group, GAPDH and RPS19 in the control group, and ACTB and GAPDH in the icv-STZ group. Additionally, we validated the normalization accuracy of the most appropriate reference genes (RPS19 and YWHAZ) by comparison with the least stable gene (TBP) using quantification of the APP and MAPT genes in the total sample group. To the best of our knowledge, this research is the first study to identify and validate the appropriate reference genes in cynomolgus monkey brains. These findings provide useful information for future studies involving the expression of target genes in the cynomolgus monkey.


SCMHBP: prediction and analysis of heme binding proteins using propensity scores of dipeptides.
Heme binding proteins (HBPs) are metalloproteins that contain a heme ligand (an iron-porphyrin complex) as the prosthetic group. Several computational methods have been proposed to predict heme binding residues and thereby to understand the interactions between heme and its host proteins. However, few in silico methods for identifying HBPs have been proposed. This work proposes a scoring card method (SCM) based method (named SCMHBP) for predicting and analyzing HBPs from sequences. A balanced dataset of 747 HBPs (selected using a Gene Ontology term GO:0020037) and 747 non-HBPs (selected from 91,414 putative non-HBPs) with an identity of 25% was firstly established. Consequently, a set of scores that quantified the propensity of amino acids and dipeptides to be HBPs is estimated using SCM to maximize the predictive accuracy of SCMHBP. Finally, the informative physicochemical properties of 20 amino acids are identified by utilizing the estimated propensity scores to be used to categorize HBPs. The training and mean test accuracies of SCMHBP applied to three independent test datasets are 85.90% and 71.57%, respectively. SCMHBP performs well relative to comparison with such methods as support vector machine (SVM), decision tree J48, and Bayes classifiers. The putative non-HBPs with high sequence propensity scores are potential HBPs, which can be further validated by experimental confirmation. The propensity scores of individual amino acids and dipeptides are examined to elucidate the interactions between heme and its host proteins. The following characteristics of HBPs are derived from the propensity scores: 1) aromatic side chains are important to the effectiveness of specific HBP functions; 2) a hydrophobic environment is important in the interaction between heme and binding sites; and 3) the whole HBP has low flexibility whereas the heme binding residues are relatively flexible. SCMHBP yields knowledge that improves our understanding of HBPs rather than merely improves the prediction accuracy in predicting HBPs.


Sarcopenic obesity: epidemiology, pathophysiology, cardiovascular disease, mortality, and management.
Sarcopenic obesity is defined as the coexistence of sarcopenia and obesity in the same individual, characterized by of the co-presence of body fat accumulation and muscle loss. This condition is currently a major concern as it is associated with frailty and disabilities such as cardiovascular disease, fractures, dementia, cancer, and increased all-cause mortality. Particularly, older individuals remain at risk of sarcopenic obesity. Progress at several levels is needed to improve the global prognostic outlook for this condition, including the elaboration and implementation of a more uniform definition that may favor the identification and specification of prevalence by age group. Furthermore, improvements in the understanding of the pathogenesis of sarcopenic obesity may lead to the development of more specific therapeutic interventions to improve prognosis. We reviewed the knowledge on sarcopenic obesity and its associations with cardiovascular diseases and mortality.


Enhanced gastric retention of solid resin beads as a marker for emetic potential of agents in rats.
Whereas nausea and emesis are burdensome side effects that lead to poor treatment compliance especially in chemotherapy, it is difficult to predict the emetic potential of agents in rats and mice because rodents do not vomit. We examined the effect of emetics on gastric retention and role of serotonin (5-hydroxytryptamine, 5-HT)<inf>3</inf> receptor in chemotherapeutic-induced enhancement of gastric retention in rats. The gastric retention of solid material was determined using resin beads, which were suitable to beads made with metals or glasses in size, hardness and weight. Each rat was orally given distilled water (0.5 ml/rat) containing 40 resin beads via a plastic feeding tube. The stomach was removed at 1 hr post-dose and cut along the greater curvature under carbon dioxide anesthesia. Beads were given immediately after administration of the drugs except with cisplatin, when there was a 1 hr delay. Cancer chemotherapeutics including cisplatin(0.1-3 mg/kg i.v.) and doxorubicin(0.3-10 mg/kg i.v.) and a nauseant, copper sulfate(1-30 mg/kg p.o.) enhanced gastric retention of beads. Ondansetron, a 5-HT<inf>3</inf> receptor antagonist, dose-dependently antagonized the enhanced gastric retention by cisplatin and doxorubicin. The copper sulfate-induced enhancement was also reversed by ondansetron. Our results suggest that 5-HT<inf>3</inf> receptors mediate the cancer chemotherapeutic-enhanced gastric retention of solid material in rats. This implicates that the gastric retention of solid material is a useful marker to predict the potential of compounds to induce nausea and/or emesis in non-vomiting rodents.


Rationale for, barriers to, and appropriate medication for the long-term treatment of depression.
Antidepressants have proven efficacy in the treatment of acute depressive episodes and the prevention of relapse over the long-term. However, whether due to ignorance about the chronicity of depression, intolerable adverse effects, or an inappropriate fear of dependence, antidepressants are often discontinued after remission or recovery from an acute episode, which frequently leads to relapse or recurrence. This, in turn, increases the risk of subsequent poor treatment response and lifelong depressive chronicity. Clinicians should focus on preventing depressive relapse with long-term antidepressant pharmacotherapy, thereby improving patients' overall outcomes, particularly with patients at high risk for relapse. When patients with depression have comorbid anxiety, benzodiazepines may be useful but should be used only as short-term augmentation during the beginning phase of antidepressant treatment; long-term treatment of comorbid anxiety is better managed by antidepressants that also treat anxiety disorders.


Simian virus 40 (SV40) and human cancer: a review of the serological data.
Serum antibodies are widely utilised as specific and sensitive markers of virus infections but they have been employed relatively infrequently in the investigation of simian virus 40 (SV40) as a human carcinogen. In the past few years, serological data have become available which allow an examination of whether SV40 is currently circulating in human communities and if SV40 infection is associated with human cancer. The development of EIA with virus-like particles (VLPs) of SV40, BKV and JCV has facilitated serological studies. Sera from macaques naturally infected with SV40 cross-react unambiguously with BKV and JCV VLPs. Tests of over 9000 human sera with different immunological assays reveal a common pattern of SV40 reactivity. A small proportion of sera react at low titers and this reactivity is unrelated to age or the geographic location of the donor, but correlates with the presence and titers of BKV and JCV antibodies. Absorption with BKV and JCV VLPs decreases or abolishes the SV40 reactivity of human sera. The SV40 reactivity of sera from patients with mesothelioma, osteosarcoma or lymphomas, cancers which are reported to be associated with SV40, was similar to that in their controls or other comparison groups. The SV40 reactivity of human sera appears to be almost entirely a result of cross-reactivity with BKV and JCV antibodies. Serological data thus do not support the possibility that SV40 is circulating in human communities or that it is associated with human cancer.


Pathology of the thyroid in severe acute respiratory syndrome.
The severe acute respiratory syndrome (SARS) epidemic started in November 2002 and spread worldwide. The pathological changes in several human organs of patients with SARS have been extensively described. However, to date, little has been reported about the effects of this infection on the thyroid gland. Femoral head necrosis and low serum triiodothyronine and thyroxine levels, commonly found in patients with SARS, raise the possibility of thyroid dysfunction. We have undertaken this study to evaluate for any potential injury to the thyroid gland caused by SARS on tissue samples obtained from 5 SARS autopsies. The terminal deoxynucleotidyl transferase-mediated dUPT nick end-labeling assay was performed to identify apoptotic cells. The follicular epithelium was found to be damaged with large numbers of cells exfoliated into the follicle. The terminal deoxynucleotidyl transferase-mediated dUPT nick end-labeling assay demonstrated many cells undergoing apoptosis. Follicular architecture was altered and showed distortion, dilatation, and collapse. No distinct calcitonin-positive cells were detectable in the SARS thyroids. In conclusion, both parafollicular and follicular cells were injured. This may provide an explanation both for low serum triiodothyronine and thyroxine levels and the osteonecrosis of the femoral head associated with patients with SARS. Apoptosis may play a role in the pathogenesis of SARS associated coronavirus infection in the thyroid gland.


Evolution of the crystal and magnetic structure of the R2MnRuO7 (R = Tb, Dy, Ho, Er, Tm, Yb, Lu, and Y) family of pyrochlore oxides.
The members of the family of pyrochlore oxides with the formula R(2)MnRuO(7) (R = Tb, Dy, Ho, Er, Tm, Yb, Lu and Y) have been synthesized and characterized. Polycrystalline samples were prepared by a soft chemistry procedure involving citrates of the different metal ions, followed by thermal treatments in air or O(2) pressure. The crystallographic and magnetic structures have been analysed from X-ray diffraction (XRD) and neutron powder diffraction (NPD) data, in complement with magnetic measurements; the evolution along the series of the crystallographic parameters (unit-cell parameters, bond distances and angles) is discussed. In R(2)MnRuO(7) pyrochlores, Mn and Ru ions statistically occupy the 16c sites in a cubic unit cell with space group Fd ̅3m, which defines an intrinsic frustrated three-dimensional system. In all the cases, the low-temperature NPD data unveils an antiferromagnetic coupling of two subsets of Mn(4+)/Ru(4+) spins, indicating that the magnetic frustration is partially relieved by the random distribution of Mn and Ru over the 16c sites. At lower temperatures there is a polarization of the R(3+) magnetic moments, which also participate in the magnetic structure, when a magnetic rare earth is present.


Severe tracheobronchial compression in a patient with Turner's syndrome undergoing repair of a complex aorto-subclavian aneurysm: anesthesia perspectives.
We present a case of severe tracheobronchial compression from a complex aorto-subclavian aneurysm in a patient with Turner's syndrome undergoing open surgical repair. Significant airway compression is a challenging situation and requires careful preoperative preparation, maintenance of spontaneous breathing when possible, and consideration of having an alternative source of oxygenation and circulation established prior to induction of general anesthesia. Cardiopulmonary monitoring is essential for safe general anesthesia and diagnosis of unexpected intraoperative events.


Frame Reflection Lab: a Playful Method for Frame Reflection on Synthetic Biology.
Synthetic biology is an emerging technology that asks for inclusive reflection on how people frame the field. To unravel how we can facilitate such reflection, this study evaluates the Frame Reflection Lab (FRL). Building upon playfulness design principles, the FRL comprises a workshop with video-narratives and co-creative group exercises. We studied how the FRL facilitated frame reflection by organizing workshops with various student groups. Analysis of 12 group conversations and 158 mini-exit surveys yielded patterns in first-order reflection (problem analysis and solution finding in reflection on the development of synthetic biology as a field) as well as patterns in second-order reflection (reflection on values and assumptions underlying the first-order reflection). Also patterns in participants' (re)framing of synthetic biology could be induced; participants' viewpoints converged to some extent, yet with openness to individual viewpoint differences. Although the FRL method fortified the reflection processes of participants, the narratives and the workshop's flexible format could inhibit the reflection too. Therefore, we advise designers of future frame reflection methods to apply stronger conversational facilitation and narratives of slightly mysterious yet identifiable narrators, in case e.g. video-narratives are created and used to scaffold the reflection process. Nevertheless, we argue that the use of a playful frame reflection method like the FRL could function well as (1) a step to precede more application-specific deliberation or decision-making on synthetic biology and as (2) a method for the collection of contemporary citizen viewpoints plus rationales underlying these, for the further (societally) responsible development of the emerging field.


A randomized trial of individual versus group-format exercise and self-management in individuals with Parkinson's disease and comorbid depression.
Depression is common in people with Parkinson's disease (PD), and exercise is known to improve depression and PD. However, lack of motivation and low self-efficacy can make exercise difficult for people with PD and comorbid depression (PD-Dep). A combined group exercise and chronic disease self-management (CDSM) program may improve the likeli-hood that individuals will engage in exercise and will show a reduction in depression symptoms. The purpose of this study was to compare changes in depression in PD-Dep between individual versus group exercise plus CDSM and to examine participant adherence and perception of the interventions. Participants (N=30) were randomized to either Enhanced EXerCisE thErapy for PD (EXCEED; group CDSM and exercise) or self-guided CDSM plus exercise. Outcomes were change in depression assessed with the Montgomery-Asberg Depression Rating Scale (MADRS), cognition, apathy, anxiety, sleep, quality of life, motor function, self-efficacy, and patient satisfaction. Both groups showed significant improvement in MADRS (P<0.001) with no significant group difference. Individuals in EXCEED group enjoyed the group dynamics but noted difficulty with the fixed-time sessions. Both group CDSM plus exercise and self-guided CDSM plus exercise can improve depression in PD-Dep. These findings suggest that development of a remotely delivered group-based CDSM format plus manualized exercise program could be useful for this population.


Rapid identification of C21 steroidal saponins in Cynanchum versicolor Bunge by electrospray ionization multi-stage tandem mass spectrometry and liquid chromatography/tandem mass spectrometry.
Electrospray ionization multi-stage tandem mass spectrometry (ESI-MSn) and liquid chromatography coupled with on-line electrospray ionization tandem mass spectrometry (LC/ESI-MSn) were performed to elucidate the clearage rule of nine investigated C21 steroidal saponins and identify them in the saponin fraction of 90% ethanolic extracts from the root and rhizome of Cynanchum versicolor Bunge. The fragments of C21 steroidal saponins in positive and negative ESI-MSn were used to deduce their mass spectral fragmentation mechanisms, and their structures were further confirmed by ESI-MSn in positive mode. The MSn spectra of the [M+Na]+ ions for saponins provided a wealth of structural information on glycosidic bond cleavage, which allowed a straightforward interpretation of spectra, with respect to the identifications of features such as the sequences of sugars attached to saponins and sugar type. By using LC/ESI-MSn, nine C21 steroidal saponins were detected in the saponin fraction of C. versicolor, and an isomer of atratoglaucoside A was elucidated simultaneously. All nine compounds showed an abundant ion for the loss of 46 Da (HCOOH) from [M+Na]+. The losses of monosaccharide sequences and aglycone as neutral fragmentation from [M+Na-HCOOH]+ were also acquired as the characteristic ions of these C21 steroidal saponins. It provided important information on monosaccharide sequences and in particular on sugar types and could be used to identify and elucidate other C21 steroidal saponins. These studies allowed us to rapidly identify C21 steroidal saponins from Radix cynanchi atrati. It is indicated that the described method had wide applicability to rapidly screen and provide structural confirmation on C21 steroidal saponins in crude materials.


Recent advances in the use of encapsulated cells for effective delivery of therapeutics.
Cell encapsulation can be defined as a living cell approach for the long-term delivery of therapeutic products. It consists of the immobilization of therapeutically active cells within a general polymer matrix that permits the ingress of nutrients and oxygen and the egress of therapeutic protein products but impedes the immune contact of the enclosed cells. In recent decades many attempts have evaluated the potential of this technology to release therapeutic agents for the treatment of different pathologies and disorders. At present, cell encapsulation may be used as a technological platform to improve knowledge and clinical use of stem cells. This review describes the main issues related to this cell-based approach and summarizes some of the most interesting therapeutic applications.


Effect of dietary polyphenol-rich grape seed on growth performance, antioxidant capacity and ileal microflora in broiler chicks.
In this study, we examined the effect of dietary supplementation with grape seed (GS) on the performance, carcass traits, plasma biochemistry, antioxidant status and ileal microflora in broilers. Experiment diets included a control diet (without additive) and three levels of GS powder (10, 20 and 40 g/kg of diet). Each diet was fed to a total of 300 one-day-old Cobb-500 chicks for 42 days. The addition of 20 g/kg of GS to the basal diet increased final body weight and body weight gain, improved the feed conversion ratio and did not affect feed intake. Dietary 20 g GS significantly increased (p < .05) the percentage of carcass yield %, dressing % and gizzard. However, the addition of 40 g/kg of GS significantly reduced the percentage of abdominal fat in the birds. Diets supplemented with GS showed the lowest content of ether extract compared with the control group (p < .05). The physical characteristics of meat and the chemical composition of DM, CP and ash were not significantly influenced by treatments. In the GS groups, plasma protein, albumin, globulin, aspartate aminotransferase and alanine aminotransferase concentrations showed no significant change compared with the control group. Broilers fed a diet supplemented with GS had lower levels of plasma glucose, total lipids, triglycerides and cholesterol compared with the control birds (p < .05). The addition of 40 g of GS significantly (p < .05) enhanced the activity of reduced glutathione, catalase, superoxide dismutase, glutathione peroxidase and GST, and correlated with significantly decreased thiobarbituric acid-reactive substances levels compared with the control group. The value of ileal pH was not significantly affected by the GS levels. Broilers fed diets supplemented with GS had lower ileal Streptococcus spp. and Escherichia coli populations but higher Lactobacillus spp. populations (p < .05). No adverse effects on birds' health were detected due to the use of GS. Thus, GS could be recommended as an herbal supplement in the diet of broiler chickens to improve performance, reduce blood lipids, enhance antioxidant capacity and decrease detrimental bacteria in the ileum.


Doping-induced memory effect in Li-ion batteries: the case of Al-doped Li4Ti5O12.
In Li-ion batteries (LIBs), a memory effect has been revealed in two-phase electrode materials such as olivine LiFePO4 and anatase TiO2, which complicates the two-phase transition and influences the estimation of the state of charge. Practical electrode materials are usually optimized by the element doping strategy, however, its impact on the memory effect has not been reported yet. Here we firstly present the doping-induced memory effect in LIBs. Pristine Li4Ti5O12 is free from the memory effect, while a distinct memory effect could be induced by Al-doping. After being discharged to a lower cutoff potential, Al-doped Li4Ti5O12 exhibits poorer electrochemical kinetics, delivering a larger overpotential during the charging process. This dependence of the overpotential on the discharging cutoff leads to the memory effect in Al-doped Li4Ti5O12. Our discovery emphasizes the impact of element doping on the memory effect of electrode materials, and thus has implications for battery design.


Hypoxia reduces HNF4α/MODY1 protein expression in pancreatic β-cells by activating AMP-activated protein kinase.
Hypoxia plays a role in the deterioration of β-cell function. Hepatocyte nuclear factor 4α (HNF4α) has an important role in pancreatic β-cells, and mutations of the human HNF4A gene cause a type of maturity-onset diabetes of the young (MODY1). However, it remains unclear whether hypoxia affects the expression of HNF4α in β-cells. Here, we report that hypoxia reduces HNF4α protein expression in β-cells. Hypoxia-inducible factor was not involved in the down-regulation of HNF4α under hypoxic conditions. The down-regulation of HNF4α was dependent on the activation of AMP-activated protein kinase (AMPK), and the reduction of HNF4α protein expression by metformin, an AMPK activator, and hypoxia was inhibited by the overexpression of a kinase-dead (KD) form of AMPKα2. In addition, hypoxia decreased the stability of the HNF4α protein, and the down-regulation of HNF4α was sensitive to proteasome inhibitors. Adenovirus-mediated overexpression of KD-AMPKα2 improved insulin secretion in metformin-treated islets, hypoxic islets, and ob/ob mouse islets. These results suggest that down-regulation of HNF4α could be of importance in β-cell dysfunction by hypoxia.


The adverse effect of FOPNL genomic variant is reversed by bortezomib-based treatment protocols in multiple myeloma.
Fibroblast growth factor receptor 1 oncogene partner N-terminal like gene (FOPNL) rs72773978 polymorphism was identified as an adverse prognostic factor in multiple myeloma (MM). We aimed to investigate the associations of rs72773978 with clinical characteristics and treatment outcome in 373 Hungarian MM patients. In our cohort, FOPNL polymorphism showed differential prognostic effect that depended on the treatment applied. Among patients treated with non-proteasome inhibitor (PI)-based therapy, carriership of the minor allele was significantly associated with adverse overall survival (p=.022). In contrast, the adverse effect was overcome by the application of PI-containing treatment (p=.048). Multivariate analyses revealed the independent adverse effect of rs72773978 on survival in the non-PI-treated group (p=.045), but not in PI treatment (OS: p=.093). We confirmed the adverse prognostic effect of rs72773978 associated with non-PI-based treatment regimens. Our results point to the importance of genotypic prognostic information associated with complex clinical background MM.


Thioredoxin-1 suppresses systemic inflammatory responses against cigarette smoking.
Thioredoxin-1 (TRX) is a small redox-active protein with antioxidative effects and redox-regulating functions. Cigarette smoking is a major etiological factor in the pathogenesis of a variety of diseases and recruits systemic immune and inflammatory responses. This report demonstrates that TRX attenuates the systemic inflammatory responses induced by cigarette smoking. The mRNA expressions of tumor necrosis factor alpha (TNF-alpha) and macrophage migration inhibitory factor (MIF) were suppressed in the spleen of TRX overexpressing transgenic mice (TRX-tg) exposed to cigarette smoking, compared with control C57BL/6 mice. In addition, protein carbonylation, a marker of cellular protein oxidation, was enhanced by cigarette smoking in the tissues of heart and liver in control mice more than in TRX-tg mice. These findings suggest that TRX may suppress the systemic inflammatory responses against cigarette smoking.


In cystic fibrosis homozygotes and heterozygotes, neutrophil apoptosis is delayed and modulated by diamide or roscovitine: evidence for an innate neutrophil disturbance.
Cystic fibrosis (CF) is a chronic inflammatory lung disease characterized by polymorphonuclear neutrophil (PMN)-dominated airway inflammation. Defective apoptosis might explain PMN persistence at these inflammation sites. We previously reported that in CF patients PMN underwent delayed apoptosis, which was not always related to their infectious state and independent of the type of CF transmembrane regulator (CFTR) mutation. To understand the role of infection and PMN apoptosis in CF, PMN apoptosis was investigated in CF parents who are obligate heterozygotes for the CFTR mutation but without chronic bacterial infection. They also demonstrated delayed PMN apoptosis compared with healthy controls, as assessed by annexin-V labeling and caspase-3 cleavage. Diamide, a direct thiol-oxidizing agent, potentiated PMN apoptosis in controls and CF patients, resulting in similar levels of constitutive and Fas-potentiated apoptosis. The cyclin-dependent kinase inhibitor roscovitine provided another approach to restore normal PMN apoptosis. However, the selective CFTR inhibitor CFTR(Inh172) did not affect PMN apoptosis in control subjects. Apparently, the dysregulation of CF PMN is not only a consequence of the chronic infectious state in CF children but might also be related to CF 'intrinsic' factors. Restoration of normal PMN apoptosis by cellular redox modulation or roscovitine opens new research avenues to decrease PMN-mediated inflammation in CF.


Frusemide administration in critically ill patients by continuous compared to bolus therapy.
Frusemide is frequently administered to critically ill patients in the intensive care unit (ICU). We investigated whether continuous frusemide infusion therapy was more effective than regular intermittent bolus doses at causing diuresis. 59 adult patients with fluid overload admitted to two tertiary multidisciplinary ICUs were randomised to either a continuous frusemide infusion or regular intermittent intravenous boluses of frusemide according to pre-defined algorithms aiming for a minimum hourly urine output. There was no significant difference in diuretic response between the two groups during the first 24 h (5.3 liters in the bolus group vs. 5.4 liters in the infusion group). In the bolus group a significantly higher dose of frusemide was needed to achieve target diuresis (24.1 vs. 9.2 mg/h in the infusion group, p = 0.0002). Mean urine output per dose of frusemide was significantly higher in the infusion group (31.6 vs. 18 ml/mg in the bolus group, p = 0.014). At the end of the study, there were no differences in hospital mortality, number of patients requiring ventilatory support, change in serum creatinine or change in estimated glomerular filtration rate. Both intermittent boluses and continuous infusion of frusemide were successful in achieving algorithm-driven diuresis. However, continuous infusion therapy was more effective than intermittent boluses since the dose of frusemide required was significantly less.


Monte Carlo skin dose simulation in intraoperative radiotherapy of breast cancer using spherical applicators.
The relatively new treatment modality electronic intraoperative radiotherapy (IORT) is gaining popularity, irradiation being obtained within a surgically produced cavity being delivered via a low-energy x-ray source and spherical applicators, primarily for early stage breast cancer. Due to the spatially dramatic dose-rate fall off with radial distance from the source and effects related to changes in the beam quality of the low keV photon spectra, dosimetric account of the Intrabeam system is rather complex. Skin dose monitoring in IORT is important due to the high dose prescription per treatment fraction. In this study, modeling of the x-ray source and related applicators were performed using the Monte Carlo N-Particle transport code. The dosimetric characteristics of the model were validated against measured data obtained using an ionization chamber and EBT3 film as dosimeters. By using a simulated breast phantom, absorbed doses to the skin for different combinations of applicator size (1.5-5 cm) and treatment depth (0.5-3 cm) were calculated. Simulation results showed overdosing of the skin (>30% of prescribed dose) at a treatment depth of 0.5 cm using applicator sizes larger than 1.5 cm. Skin doses were significantly increased with applicator size, insofar as delivering 12 Gy (60% of the prescribed dose) to skin for the largest sized applicator (5 cm diameter) and treatment depth of 0.5 cm. It is concluded that the recommended 0.5-1 cm distance between the skin and applicator surface does not guarantee skin safety and skin dose is generally more significant in cases with the larger applicators. • Intrabeam x-ray source and spherical applicators were simulated and skin dose was calculated. • Skin dose for constant skin to applicator distance strongly depends on applicator size. • Use of larger applicators generally results in higher skin dose. • The recommended 0.5-1 cm skin to applicator distance does not guarantee skin safety.


Human plasma metabolomics in age-related macular degeneration (AMD) using nuclear magnetic resonance spectroscopy.
To differentiate the plasma metabolomic profile of patients with age related macular degeneration (AMD) from that of controls, by Nuclear Magnetic Resonance (NMR) spectroscopy. Two cohorts (total of 396 subjects) representative of central Portugal and Boston, USA phenotypes were studied. For each cohort, subjects were grouped according to AMD stage (early, intermediate and late). Multivariate analysis of plasma NMR spectra was performed, followed by signal integration and univariate analysis. Small changes were detected in the levels of some amino acids, organic acids, dimethyl sulfone and specific lipid moieties, thus providing some biochemical information on the disease. The possible confounding effects of gender, smoking history and age were assessed in each cohort and found to be minimal when compared to that of the disease. A similar observation was noted in relation to age-related comorbidities. Furthermore, partially distinct putative AMD metabolite fingerprints were noted for the two cohorts studied, reflecting the importance of nutritional and other lifestyle habits in determining AMD metabolic response and potential biomarker fingerprints. Notably, some of the metabolite changes detected were noted as potentially differentiating controls from patients diagnosed with early AMD. For the first time, this study showed metabolite changes in the plasma of patients with AMD as compared to controls, using NMR. Geographical origins were seen to affect AMD patients´ metabolic profile and some metabolites were found to be valuable in potentially differentiating controls from early stage AMD patients. Metabolomics has the potential of identifying biomarkers for AMD, and further work in this area is warranted.


Dendrite-free deposition on lithium anode toward long-life and high-stable Li//graphite dual-ion battery.
Metallic lithium protection plays a crucial role on improving the electrochemical properties of Li-anode-based batteries. Herein, for an advanced Li//graphite dual-ion battery, constructing a robust and conductive film of carbon nanofibers on a Li anode effectively achieves dendrite-free Li growth and hence significantly enhances the long-life cyclic stability.


[Endoscopic intraluminal valvuloplasty--a therapy of the future for gastroesophageal reflux].
A new endoscopic intraluminal valvuloplasty is described. The procedure provides a simple, easy out-patient approach for antireflux surgery and is applicable to patients with early gastroesophageal reflux disease as an alternative to chronic life-long medical therapy. The feasibility, durability and efficacy of the procedure in baboons are reported.


O-fucose modulates Notch-controlled blood lineage commitment.
Notch receptors are cell surface molecules essential for cell fate determination. Notch signaling is subject to tight regulation at multiple levels, including the posttranslational modification of Notch receptors by O-linked fucosylation, a reaction that is catalyzed by protein O-fucosyltransferase-1 (Pofut1). Our previous studies identified a myeloproliferative phenotype in mice conditionally deficient in cellular fucosylation that is attributable to a loss of Notch-dependent suppression of myelopoiesis. Here, we report that hematopoietic stem cells deficient in cellular fucosylation display decreased frequency and defective repopulating ability as well as decreased lymphoid but increased myeloid developmental potential. This phenotype may be attributed to suppressed Notch ligand binding and reduced downstream signaling of Notch activity in hematopoietic stem cells. Consistent with this finding, we further demonstrate that mouse embryonic stem cells deficient in Notch1 (Notch1(-/-)) or Pofut1 (Pofut1(-/-)) fail to generate T lymphocytes but differentiate into myeloid cells while coculturing with Notch ligand-expressing bone marrow stromal cells in vitro. Moreover, in vivo hematopoietic reconstitution of CD34(+) progenitor cells derived from either Notch1(-/-) or Pofut1(-/-) embryonic stem cells show enhanced granulopoiesis with depressed lymphoid lineage development. Together, these results indicate that Notch signaling maintains hematopoietic lineage homeostasis by promoting lymphoid development and suppressing overt myelopoiesis, in part through processes controlled by O-linked fucosylation of Notch receptors.


Gene Expression and Microcomputed Tomography Analysis of Grafted Bone Using Deproteinized Bovine Bone and Freeze-Dried Human Bone.
Bio-Oss and demineralized freeze-dried bone allograft (DFDBA) are two commercial bone grafts that have been associated with clinical success for many years. However, there are few in vivo studies regarding their healing mechanism. The purpose of this study was to investigate the level of bone formation using microcomputed tomography (micro-CT) and gene expression in mouse calvaria at 1 and 3 months after bone grafting with deproteinized bovine bone and freeze-dried human bone, and compare them to natural bone healing. Thirty-six mice were divided into three groups (n = 6 per group) according to the type of bone graft used: group 1 (control)-an empty defect without bone graft; group 2-treatment with deproteinized bovine xenograft (Bio-Oss); group 3-treatment with DFDBA. The bone graft was inserted into two 3-mm calvarial defects created on both sides of the parietal bone. At 1 and 3 months, the mice were sacrificed and bone volume was evaluated using micro-CT and gene expression analysis using reverse transcription polymerase chain reaction (RT-PCR). Micro-CT analysis demonstrated that the parietal bone of mice grafted with Bio-Oss had significantly greater bone volume than both the DFDBA and control groups at both 1 and 3 months. The expression of bone marker genes (Runx2, Osterix [Osx], alkaline phosphatase [ALP], osteopontin [OPN], and osteocalcin [OCN]) were significantly increased from 1 month in both Bio-Oss and DFDBA groups at 3 months. Runx2 and Osx had significantly higher expression in the Bio-Oss and DFDBA groups compared to the control group at 3 months. No statistically significant difference was observed among groups after 1 month. These results showed that both bone graft materials promoted bone regeneration. Bio-Oss demonstrated high osteoconductive properties.


Altered paracrine signaling from the injured knee joint impairs postnatal long bone growth.
Regulation of organ growth is a poorly understood process. In the long bones, the growth plates (GPs) drive elongation by generating a scaffold progressively replaced by bone. Although studies have focused on intrinsic GP regulation, classic and recent experiments suggest that local signals also modulate GP function. We devised a genetic mouse model to study extrinsic long bone growth modulation, in which injury is specifically induced in the left hindlimb, such that the right hindlimb serves as an internal control. Remarkably, when only mesenchyme cells surrounding postnatal GPs were killed, left bone growth was nevertheless reduced. GP signaling was impaired by altered paracrine signals from the knee joint, including activation of the injury response and, in neonates, dampened IGF1 production. Importantly, only the combined prevention of both responses rescued neonatal growth. Thus, we identified signals from the knee joint that modulate bone growth and could underlie establishment of body proportions.


Drotrecogin alfa (activated) approved for treatment of severe sepsis.
Drotrecogin alfa (activated) is a recombinant form of human activated protein C that has been shown to reduce mortality in severely septic patients with a high risk of death. Given the complexity of the inclusion and exclusion criteria, the stability and unique duration of infusion, and the cost of treatment, therapy with this agent should be carefully considered to ensure its appropriate use.


Sudden death after open gastric bypass surgery.
Gastric bypass surgery has become a relatively low-risk bariatric surgical intervention in a high-risk patient population (Nguyen et al., Arch Surg, 141:445-449, 2006; Buchwald et al. JAMA, 13:1724-1737, 2004). Surgical interventions in patients suffering from morbid obesity are typically associated with excess morbidity (Parikh et al., Am Surg, 73:959-962, 2007). Though overall mortality after bariatric surgery is <1% is low (Mason et al., Obes Surg, 17:9-14, 2007), some surgical complications such as anastomotic leaks, staple line disruption and bowel obstruction may still impact on postoperative outcome (Parikh et al., Am Surg, 73:959-962, 2007; Mason et al., Obes Surg, 17:9-14, 2007). Early symptoms are often missed, as clinical presentation may be discreet, inexistent or falsely attributed to obesity. This case report refers to a patient in whom discomfort and agitation associated with a rise in temperature heralded a fulminant septic shock syndrome precipitating his death. Literature on early complications and management after gastric bypass is reviewed. A high level of suspicion should be present in the case of an unexpected postoperative deterioration of the patient's general condition. Time to treat may be very short (Mason et al., Obes Surg, 17:9-14, 2007). Computed tomography is mandatory to rule out pulmonary embolism and bypass obstruction.


Sensitivity to binaural intensity and phase difference cues in kitten inferior colliculus.
1. Responses of single neurons to monaural or binaural CF tones delivered through a closed and calibrated sound delivery system were studied in the central nucleus of the inferior colliculus (ICC) in ketamine and barbiturate-anesthetized kittens 4-105 days old. 2. Neurons from young kittens had elevated thresholds, some greater than 100 dB in the youngest kittens tested. Average thresholds in the ICC matched those previously measured in the auditory nerve (AN), cochlear nuclei (CN), and auditory cortex (CTX), suggesting that the drop in threshold as a function of age is primarily determined by development at the periphery. 3. Minimal first-spike latencies were relatively long in the youngest kittens, approaching adult values by the end of the third postnatal week. Latencies were distributed between values previously determined for the CN and auditory cortex and can be attributed to the centripetal development of the auditory system. 4. The range of frequencies that were effective in exciting ICC neurons was restricted in young kittens. Neurons having characteristic frequencies (CFs) greater than 7 kHz were not recorded before postnatal day 10. CF distribution matched that obtained in previous experiments from AN, CN, and CTX, reflecting the development of the cochlea. 5. Both monotonic and nonomonotonic spike count-versus-intensity functions were found in the youngest kittens. There was a tendency for monotonic functions from the youngest kittens to be steeper than those from older kittens. No age-related changes in the shapes of non-monotonic functions were found. 6. Sensitivity to interaural intensity difference (IID), tested by holding the intensity to the excitatory ear at a suprathreshold level and increasing the intensity of the stimulus to the inhibitory ear, was exhibited as early as 8 days after birth. The majority of the cells exhibiting sensitivity to IID (89.5%) were classified as EI cells, and almost all IID sensitive cells had CFs between 3 and 25 kHz. Within our sample the shapes of IID functions, as well as the operating range of the IID functions, closely resembled those obtained from the adult cat. Thresholds of excitation and of inhibition were highly correlated, suggesting that the ipsilateral and contralateral inputs to the ICC develop as a matched set. 7. Sensitivity to interaural phase difference (IPD), tested either by shifting the onset phase of a CF tone to one ear relative to the other or by presenting tones of slightly different frequency to the two ears, was present as early as 12 days after birth.(ABSTRACT TRUNCATED AT 400 WORDS)


Pathogenetic mechanisms and treatment targets in cerebral malaria.
Malaria, the most prevalent mosquito-borne infectious disease worldwide, has accompanied humanity for millennia and remains an important public health issue despite advances in its prevention and treatment. Most infections are asymptomatic, but a small percentage of individuals with a heavy parasite burden develop severe malaria, a group of clinical syndromes attributable to organ dysfunction. Cerebral malaria is an infrequent but life-threatening complication of severe malaria that presents as an acute cerebrovascular encephalopathy characterized by unarousable coma. Despite effective antiparasite drug treatment, 20% of patients with cerebral malaria die from this disease, and many survivors of cerebral malaria have neurocognitive impairment. Thus, an important unmet clinical need is to rapidly identify people with malaria who are at risk of developing cerebral malaria and to develop preventive, adjunctive and neuroprotective treatments for cerebral malaria. This Review describes important advances in the understanding of cerebral malaria over the past two decades and discusses how these mechanistic insights could be translated into new therapies.


New microsporidia parasitizing bark lice (Insecta: Psocoptera).
Two species of bark lice, Xanthocaecilius sommermanae Mockford and Polypsocus corruptus Hagen, collected in a canopy Malaise trap placed in Great Smoky Mountains National Park as part of a survey of the park's fauna, were found to be infected with microsporidia. Diagnosis was originally based on light microscopy, and was confirmed by PCR amplification and electron microscopy. This is the first record of microsporidia infection in the insect order Psocoptera. Four morphological spore types corresponded to four original SSUrDNA sequences (Genbank accession no. FJ865221-24), suggesting infection with four microsporidia species. Two of those species were examined by electron microscopy. We describe here one new genus and two new species based on morphological and sequence data: Antonospora psocopterae sp. n. with elongated diplokaryotic spores, 4.4+/-0.05 x 1.9+/-0.03 microm and Mockfordia xanthocaeciliae gen. n. sp. n. with ovocylindrical monokaryotic spores, 2.5+/-0.10 x 1.4+/-0.02 microm. A. psocopterae displayed high sequence (95%) and structural similarity with Antonospora scoticae, fell within a well supported dichotomy with A. scoticae inside the Antonospora-Paranosema clade in phylogenetic analyses by NJ, PS and ML. M. xanthocaeciliae did not exhibit much sequence or structural similarity with any of known microsporidia species, except Encephalitozoon spp. M. xanthocaeciliae fell within one clade with Encephalitozoon spp. in phylogenies and shared with encephalitozoons structural resemblance and about 80% of SSUrDNA sequence identity. The other two species were not described and provisionally were placed to the collective genus Microsporidium as Microsporidium sp. 1 and Microsporidium sp. 4 from bark lice because of insufficient morphological data. The finding that samples fixed and stored for months in propylene glycol ("antifreeze") are good enough for DNA sequence analysis and can be used for morphological analyses (if no better fixation alternatives are available), is promising for future surveys for microsporidia.


Long term results of percutaneous fixation of proximal humerus fractures.
Proximal humerus fracture in elderly osteoporotic patients usually leads to severe displaced and multifragmentary fractures. Associated comorbidities may limit surgical options and conservative treatment is commonly indicated, however, with variable results. In most cases, surgery is the treatment of choice in order to restore anatomical integrity, and allow early functional recovery. Several techniques were used over the years, each with specific indication. Percutaneous pinning after closed reduction, a mini-invasive technique and fixation by use of K-wires is not preferred commonly. We present our experience with this approach, focusing on its indications and advantages. A study group of 41 consecutive patients with a mean age of 65.5 years were evaluated clinically (VAS, Constant-Murley score, range of motion), and with radiological analysis: 35 patients finally completed a minimum followup of 24 months. K-wires were removed after a mean interval of 4 weeks. Clinical and radiographic healing occurred in a mean time of 8.2 weeks in all fractures but one, with improvements in mean Constant-Murley score of 87.6 points, mean VAS of 2.3. In 33 patients, the reduction was considered satisfactory. In two cases, reduction was poor, but the patients however presented acceptable functional outcome. Percutaneous pinning may represent a suitable option of treatment for 2-or 3-part proximal humerus fractures in selected subjects.


Stemming the hype: what can we learn from iPSC models of Parkinson's disease and how can we learn it?
Parkinson's disease (PD) is a prevalent and debilitating neurodegenerative disorder for which there are no available cures. PD pathogenesis is poorly understood because appropriate animal and in vitro models are lacking. The development of induced Pluripotent Stem Cells (iPSCs) has allowed researchers to generate disease-specific dopaminergic neurons in vitro by reprogramming skin cells from patients with the disease. It is hoped that this unprecedented access to PD patients' neurons will yield mechanistic insights into PD pathogenesis, a platform for drug screening, and a means of early diagnosis. In this article I critically evaluate the current usage of iPSCs in PD research. I first outline the iPSC paradigm and emphasise the benefits of this approach for modelling PD. I then ask what we can learn from the iPSC-based studies done to date. I argue that these studies have not been particularly informative when considered as an isolated body of evidence. I suggest that the limitations of this technology can be overcome, and I conclude that iPSCs have the potential to be an extremely useful tool in PD research. However, they will never be a panacea and should continue to be used in concert with other in vitro and animal models.


Success after treatment of periprosthetic joint infection: a Delphi-based international multidisciplinary consensus.
The lack of agreement regarding what constitutes successful treatment for periprosthetic joint infections (PJI) makes it difficult to compare the different strategies of management that are used in clinical practice and in research studies. The aims of this study were to create a consensus definition for success after PJI treatment, and to provide a universal, multidimensional framework for reporting of studies regarding PJI treatment. A two-round basic Delphi method was used to reach a consensus definition. We invited 159 international experts (orthopaedic surgeons, infectious disease specialists, and clinical researchers) from 17 countries to participate; 59 participated in the first round, and 42 participated in the second round. The final definition consisted of all statements that achieved strong agreement (80% or greater of participants considering a criterion relevant for defining success). The consensus definition of a successfully treated PJI is: (1) infection eradication, characterized by a healed wound without fistula, drainage, or pain, and no infection recurrence caused by the same organism strain; (2) no subsequent surgical intervention for infection after reimplantation surgery; and (3) no occurrence of PJI-related mortality (by causes such as sepsis, necrotizing fasciitis). The Delphi panel agreed to defining midterm results as those reported 5 or more years after the definitive PJI surgery, and long-term results as those reported 10 or more years after surgery. Although no consensus was reached on the definition of short-term results, 71% of the participants agreed that 2 years after the definitive PJI surgery is acceptable to define it. This multidimensional definition of success after PJI treatment may be used to report and compare results of treatment of this catastrophic complication. Level V, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence.


Activity of sulfamethoxazole and trimethoprim against Bacteroides fragilis.
Minimum inhibitory concentrations (MICs) of sulfamethoxazole (SMX) and trimethoprim (TMP), alone and in three combinations, 20:1, 1:1, and 1:20, were determined on Diagnostic Sensitivity Test (DST) and Mueller-Hinton (MH) agars containing lysed blood for various inocula of 91 strains of Bacteroides fragilis from the U.S.A. and U.K. MICs of SMX were high with large inocula and higher on MH than DST, but results for TMP were less affected by these two factors. True SMX resistance was rare: 10 U.S.A. strains previously reported as resistant appeared to be susceptible. Maximum potentiation of MICs was observed when SMX and TMP were combined in ratios close to those of the ratios of their MICs, that is, SMX/TMP 20:1 for large inocula and the reverse for small inocula for determinations on DST and usually 20:1 for all inoculum sizes on MH. These observations explain some of the discrepancies in reports, but defer the problem of potential usefulness of the drugs in the treatment of infection with anaerobes to future study.


Low number of invariant NKT cells is associated with poor survival in acute myeloid leukemia.
T lymphocytes play an important role in the immunosurveillance of patients with hematologic malignancies. No study has so far examined the association between the number of lymphocyte subsets at diagnosis and overall survival (OS). We examined this relationship in adult patients with de novo acute myeloid leukemia (AML). A longitudinal prospective study was conducted on 28 AML patients. Peripheral blood (PB) and bone marrow (BM) were obtained before chemotherapy to quantify the number of CD4+ and CD8+ T cells, natural killer (NK), invariant NKT (iNKT), and type-1 and type-2 dendritic cells. The Kaplan-Meier and Cox proportional hazard model were used to determine significant association between the number of each cell subtype and survival. BM counts of CD4+ lymphocytes >506.11/μL and CD8+ T lymphocytes >556.02/μL and a PB count of iNKT cells <0.2/μL were associated with poor OS by univariate analysis (P = 0.015, P = 0.009, P = 0.033 respectively). Multivariate analysis showed that an iNKT count <0.2 cells/μL is an independent prognostic factor for OS. An iNKT cell number of <0.2/μL confers a poor prognosis in de novo AML patients.


Liver-specific ablation of Krüppel-associated box-associated protein 1 in mice leads to male-predominant hepatosteatosis and development of liver adenoma.
The liver is characterized by sexually dimorphic gene expression translating into sex-specific differences in lipid, drug, steroid hormone, and xenobiotic metabolism, with distinct responses of males and females to environmental challenges. Here, we investigated the role of the Krüppel-associated box (KRAB)-associated protein 1 (KAP1) epigenetic regulator in this process. Liver-specific KAP1 knockout (KO) led to strikingly sexually dimorphic phenotypic disturbances, including male-predominant steatosis and hepatic tumors with up-regulation of protein kinase B and extracellular signal-related kinases 1/2 mitogen-activated protein kinase signaling. This correlated with the sex-specific transcriptional dysregulation of a wide range of metabolic genes, notably those involved in retinol and sex hormone processing as well as in detoxification. Furthermore, chromatin immunoprecipitation followed by deep sequencing indicated that a number of dysregulated genes are direct targets of the KRAB/KAP1 repression system. Those genes include sexually dimorphic cytochrome P 450 Cyp2d9, glutathione S-transferase π, Cyp2a, Cyp2b, and Cyp3a gene clusters. Additionally, we identified a male-restricted KAP1-binding site in the fat-specific protein 27 gene, correlating with its male-predominant up-regulation upon Kap1 deletion, suggesting that the latter might be an important trigger in the development of male-specific hepatosteatosis and secondary tumorigenesis. This work reveals KRAB/KAP1-mediated transcriptional regulation as a central event in metabolic control hormones, drugs, and xenobiotics in the liver and further links disturbances in these processes with hepatic carcinogenesis.


Alpha 1A-adrenoceptors in rabbit bronchus.
In the presence of propranolol, bronchial preparations from rabbits were markedly contracted by norepinephrine and phenylephrine but only slightly by clonidine. The contractile responses to norepinephrine were inhibited by prazosin and by high concentrations of yohimbine. The concentration-response curve of norepinephrine was shifted in parallel by WB4101 and 5-methylurapidil but not by 60-min treatment with chloroethylclonidine. These results suggest strongly that norepinephrine-induced contraction of rabbit bronchus is mediated through alpha 1A-adrenoceptors. Furthermore, cyclooxygenase inhibitors did not influence the response to norepinephrine, so norepinephrine-induced contraction is considered not to be mediated through the release of prostaglandins.


Acoustic and dynamic mechanical properties of a polyurethane rubber.
Acoustical and dynamic mechanical measurements were carried out on a commercial polyurethane rubber, DeSoto PR1547. The sound speed and attenuation were measured over the range from 12.5 to 75 kHz and 3.9 to 33.6 degrees C. Shear modulus was measured from 10(-4) to 2 Hz and -36 to 34 degrees C. The peak heights of the shear loss tangent varied with temperature, demonstrating thermorheological complexity. At higher temperatures, time-temperature superpositioning could be applied, with the shift factors following the Williams-Landel-Ferry equation. From the combined acoustical and mechanical measurements, values for the dynamic bulk modulus were determined. Moreover, superposition of the bulk modulus data was achieved using the shift factors determined from the dynamic mechanical shear measurements. Finally, this work illustrates the capability and the working rules of acoustical measurements in a small tank.


Senescence marker protein 30 has a cardio-protective role in doxorubicin-induced cardiac dysfunction.
Senescence marker protein 30 (SMP30), which was originally identified as an aging marker protein, is assumed to act as a novel anti-aging factor in the liver, lungs and brain. We hypothesized that SMP30 has cardio-protective function due to its anti-aging and anti-oxidant effects on doxorubicin (DOX)-induced cardiac dysfunction. SMP30 knockout (SMP30 KO) mice, SMP30 transgenic (SMP30 TG) mice with cardiac-specific overexpression of SMP30 gene and wild-type (WT) littermate mice at 12-14 weeks of age were given intra-peritoneal injection of DOX (20 mg/kg) or saline. Five days after DOX injection, echocardiography revealed that left ventricular ejection fraction was more severely reduced in the DOX-treated SMP30 KO mice than in the DOX-treated WT mice, but was preserved in the DOX-treated SMP30 TG mice. Generation of reactive oxygen species and oxidative DNA damage in the myocardium were greater in the DOX-treated SMP30 KO mice than in the DOX-treated WT mice, but much less in the SMP30 TG mice. The numbers of deoxynucleotidyltransferase-mediated dUTP nick end-labeling positive nuclei in the myocardium, apoptotic signaling pathways such as caspase-3 activity, Bax/Bcl-2 ratio and phosphorylation activity of c-Jun N-terminal kinase were increased in SMP30 KO mice and decreased in SMP30 TG mice compared with WT mice after DOX injection. SMP30 has a cardio-protective role by anti-oxidative and anti-apoptotic effects in DOX-induced cardiotoxicity, and can be a new therapeutic target to prevent DOX-induced heart failure.


Non-invasive temperature mapping using temperature-responsive water saturation shift referencing (T-WASSR) MRI.
We present a non-invasive MRI approach for assessing the water proton resonance frequency (PRF) shifts associated with changes in temperature. This method is based on water saturation shift referencing (WASSR), a method first developed for assessing B0 field inhomogeneity. Temperature-induced water PRF shifts were determined by estimating the frequency of the minimum intensity of the water direct saturation spectrum at each temperature using Lorentzian line-shape fitting. The change in temperature was then calculated from the difference in water PRF shifts between temperatures. Optimal acquisition parameters were first estimated using simulations and later confirmed experimentally. Results in vitro and in vivo showed that the temperature changes measured using the temperature-responsive WASSR (T-WASSR) were in good agreement with those obtained with MR spectroscopy or phase-mapping-based water PRF measurement methods,. In addition, the feasibility of temperature mapping in fat-containing tissue is demonstrated in vitro. In conclusion, the T-WASSR approach provides an alternative for non-invasive temperature mapping by MRI, especially suitable for temperature measurements in fat-containing tissues.


The impact of organic carbon on the performance of a high rate nitrifying trickling filter designed to pre-treat potable water.
The application of nitrifying trickling filters (NTFs) to potable water treatment is less well understood than their application to wastewater treatment, particularly regarding the effect of low ammonia substrate concentrations and organic carbon loading on filter performance. A large pilot-scale NTF was operated under conditions that simulated the raw water quality of poorly protected catchments typically found in SE Asia, with the objective of reducing the ammonia driven chlorine demand during disinfection. The efficacy of a high rate NTF to remove low concentrations of ammonia (0.5-5.0 mg NH(4)-N L(-1)) in the presence of high organic carbon (1-12 mg soluble biochemical oxygen demand (sBOD(5)) L(-1)) was investigated. Results demonstrated that 90 to 100% of nitrification was maintained only when the carbon load was less than 0.7 g sBOD(5) m(-2) d(-1) (<4 mg sBOD(5) L(-1)). Once the organic load was increased beyond 0.75 to 2.1 g sBOD(5) m(-2) d(-1) (4.5-12.1 mg sBOD(5) L(-1)), a linear decline in nitrification from 70 to 15% was observed within a timeframe of 8 to 10 d. The impact of high organic loads on the distribution of nitrification down the NTF was also investigated. Results confirmed that carbon loads greater than 0.95 g sBOD(5) m(-2) d(-1) (>5.5 mg sBOD(5) L(-1)), severely suppressed nitrification throughout the entire filter bed.


Lived experience of being a nurse from a male and female perspective.
The phenomenological psychology approach was used here to encourage reflection on the experience of nursing from both male and female perspectives, then to explore and analyse the experience and compare and contrast the findings with current literature. The research question being explored was 'Is the lived experience of being a nurse different depending on your gender?' Using individual semi-structured interviews, questions were asked of two participants. These interviews were transcribed, analysed using an interpretative or 'hermeneutic' approach to identify main themes and, together with reflexive field notes written during the study, used to complete a qualitative analysis. The main themes identified in the analysis of the transcripts were intersubjectivity, career versus vocation and gender stereotyping. The experience expressed by the two research participants suggests that the gender of a nurse influences the lived experience when compared and contrasted. As no previous gender comparative research could be found during the background literature search, this study is novel.


Interbody lumbar fusion using a carbon fiber cage implant versus allograft bone. An investigational study in the Spanish goat.
A carbon fiber-reinforced polymer implant, designed to aid interbody lumbar fusion, was tested biologically in an experimental surgical model. Twenty-seven Spanish goats had interbody lumbar fusion surgery in a randomized protocol. Seventeen goats were implanted with the carbon fiber-reinforced polymer cage packed with autologous bone, and 10 goats were implanted with ethylene oxide-sterilized allograft bone. To determine fusion success, biocompatibility of the carbon polymer material, and possibility of carbon wear debris at intervals after surgical implantation. Goats were killed at 6 months, 12 months, and 24 months and full-body autopsies were done. Spine specimens were studied by plain radiography, three-dimensional reformatted computed tomography studies, and histology. At 6 months, one of three allograft implantations showed histologic and radiographic fusion, whereas five of five carbon fiber-reinforced polymer cage fusions showed at least partial fusion. At 12 months, two of three allograft implantations and five of five carbon fiber-reinforced polymer cage fusions were solidly fused. At 24 months, five of five allograft implantations and three of three carbon fiber-reinforced polymer cage implantations were solidly fused. Interbody fusion using a carbon cage implant packed with autologous bone achieved a quicker and more reliable fusion compared with ethylene oxide-sterilized allograft bone. There were no adverse effects from the implant material.


Protective immunity against lethal F. tularensis holarctica LVS provided by vaccination with selected novel CD8+ T cell epitopes.
Recently we described an unbiased bacterial whole-genome immunoinformatic analysis aimed at selection of potential CTL epitopes located in "hotspots" of predicted MHC-I binders. Applying this approach to the proteome of the facultative intra-cellular pathogen Francisella tularensis resulted in identification of 170 novel CTL epitopes, several of which were shown to elicit highly robust T cell responses. Here we demonstrate that by DNA immunization using a short DNA fragment expressing six of the most prominent identified CTL epitopes a potent and specific CD8+ T cell responses is being induced, to all encoded epitopes, a response not observed in control mice immunized with the DNA vector alone Moreover, this CTL-specific mediated immune response prevented disease development, allowed for a rapid clearance of the bacterial infection and provided complete protection against lethal challenge (10LD50) with F. tularensis holarctica Live Vaccine Strain (LVS) (a total to 30 of 30 immunized mice survived the challenge while all control DNA vector immunized mice succumbed). Furthermore, and in accordance with these results, CD8 deficient mice could not be protected from lethal challenge after immunization with the CTL-polyepitope. Vaccination with the DNA poly-epitope construct could even protect mice (8/10) against the more demanding pulmonary lethal challenge of LVS. Our approach provides a proof-of-principle for selecting and generating a multi-epitpoe CD8 T cell-stimulating vaccine against a model intracellular bacterium.


Calcium-activated protease in hamster cardiomyopathy.
A high calcium-requiring protease was purified from the hearts of myopathic hamsters. The biochemical properties of the enzyme were studied with [3H]acetylcasein as substrate. Comparison of the enzyme from hamster and rat hearts indicated no species specificity. Increased levels of the enzyme were associated with the development of cardiac lesions in myopathic hamsters.


Learning from error: leading a culture of safety.
A recent shift towards more collective leadership in the NHS can help to achieve a culture of safety, particularly through encouraging frontline staff to participate and take responsibility for improving safety through learning from error and near misses. Leaders must ensure that they provide psychological safety, organizational fairness and learning systems for staff to feel confident in raising concerns, that they have the autonomy and skills to lead continual improvement, and that they have responsibility for spreading this learning within and across organizations.


Recombinant protein to analyze autoantibodies to proteinase 3 in systemic vasculitis.
The presence of antineutrophil cytoplasmic autoantibodies with specificity for proteinase 3 (PR3-ANCA) usually is detected by enzyme-linked immunosorbent assay (ELISA) with purified PR3 as a substrate. We studied the technical performance of direct and capture ELISA using a recombinant proteolytically inactive form of PR3 produced in the baculovirus expression system for the detection of PR3-ANCA in 114 patients with systemic vasculitis at diagnosis. We found that ELISA using recombinant PR3 produced in insect cells is a promising alternative for ELISA with native PR3. We found a correlation between tests using recombinant or native PR3, as well as correlation of the ELISA results with ANCA titers measured by the indirect immunofluorescence technique. However, the specificity for ANCA-associated vasculitis of ELISA with recombinant PR3 was lower than ELISA using native PR3. Compared with the direct assay, capture ELISA is a more sensitive method for PR3-ANCA detection, with both native and recombinant PR3, and its results depend on the monoclonal antibody used to capture the antigen.


The impact of endoscopic third ventriculostomy on the management of newly diagnosed hydrocephalus in infants.
To evaluate the role of endoscopic third ventriculostomy (ETV) as a primary treatment for hydrocephalus in children less than 1 year old and to determine its impact as a whole on the reduction of shunts necessary in a new population of hydrocephalic infants. Data were collected prospectively on 47 infants with newly diagnosed hydrocephalus of all aetiologies who were referred between 1st April 1998 and 30th September 2000. Twenty-one patients (median age 6 weeks, range 34 weeks of gestation to 10 months) underwent ETV, while the remaining 26 patients had insertion of a ventriculoperitoneal shunt. Anatomical criteria and demonstration of third ventricle outflow obstruction on preoperative magnetic resonance imaging were used to select patients for ETV. There was no mortality or major morbidity following ETV. The median follow-up period was 18 (range 8-36) months. During the follow-up period, the ETV remained patent in 7 (33%) of the 21 patients. Of the 14 patients with failed ETV, 11 had insertion of a ventriculoperitoneal shunt, while 3 have undergone successful redo ETV. Therefore, in total 10 patients (48%) of the ETV group remain shunt independent. The best results were obtained in patients with congenital aqueduct stenosis with 71% (5 of 7 patients) success rate, while patients with posthaemorrhagic hydrocephalus did particularly badly with only 1 of 10 patients having a successful ETV. Overall, 10 of 47 (21%) infants with newly diagnosed hydrocephalus have avoided a shunt. Our results suggest that the selective use of ETV as the primary treatment in infants with hydrocephalus is safe and can lead to a reduction in the shunted population of all newly diagnosed hydrocephalic infants by up to 21%. Success of ETV is aetiology, not age dependent.


Herpes simplex virus type 1 and respiratory disease in critically-ill patients: Real pathogen or innocent bystander?
Herpes simplex virus type 1 (HSV-1) has been associated with pulmonary disease, mostly in severely immunocompromised patients. After reactivation and shedding in the oropharynx, the virus may reach the lower respiratory tract by aspiration or by contiguous spread. HSV-1 can be detected in clinical specimens by virus culture or quantitatively by nucleic acid amplification techniques. With these techniques, HSV-1 is often detected in the respiratory secretions of critically-ill patients. However, a clear diagnosis of HSV-1 pneumonia is difficult to establish because clinical criteria, radiological features and laboratory findings all lack specificity. Lower respiratory tract HSV-1 infections have not been associated with specific risk-factors. There is also an absence of consistent data concerning the effect of antiviral treatment on the outcome of critically-ill patients. Further studies are needed to better define the pathogenic role of HSV-1 in the lower respiratory tract of these patients, to improve the diagnosis, and, especially, to assess the need for antiviral treatment in the individual patient.


A new method to detect acrosome-reacted spermatozoa using biotinylated soybean trypsin inhibitor.
To develop a method to detect acrosome-reacted spermatozoa on human zonae pellucidae using only commercially available reagents and without need for sperm fixation. Sperm head labeling with biotinylated soybean trypsin inhibitor (SBTI-biotin) was compared with results of a known method using fluorescein isothiocyanate (FITC)-conjugated Pisum sativum agglutinin. The SBTI-biotin method was applied to sperm bound to human zonae pellucidae. Healthy sperm donors with normal semen characteristics were recruited by the Laboratory for Reproductive Medicine in a university medical center. Soybean trypsin inhibitor-biotin binding patterns on nonfixed spermatozoa were visualized with avidin-Texas Red. The development in time of various patterns upon induction of acrosome reaction (AR) in suspension with Ca(2+)-ionophore A23187 was noted and compared with P. sativum agglutinin FITC labeling patterns. Soybean trypsin inhibitor-biotin labeling patterns of spermatozoa bound to human zonae pellucidae were determined. Soybean trypsin inhibitor-biotin bound specifically, via the SBTI-moiety, to an acrosomal factor as soon as AR started. Sperm-head labeling patterns could be assigned to defined stages of the AR process. The results were highly correlated to those obtained with P. sativum agglutinin FITC. The end point of the AR in suspension and on zonae pellucidae was SBTI-biotin binding confined to the equatorial segment. The SBTI-biotin method can be used to detect nonfixed acrosome-reacted spermatozoa both in suspension and on zonae pellucidae.


Immunologic characteristics of the leukocyte adherence inhibition assay in rheumatoid arthritis.
Peripheral blood leukocytes (PBL) from patients with rheumatoid arthritis (RA) when used in the leukocyte adherence inhibition (LAI) assay were capable of distinguishing antigenic differences between RA and osteoarthritic (OA) synovial membrane extracts. Normal PBL from control subjects with nonadherence index (NAI) values of 7 +/- 2 were able to respond positively (NAI values of 31 +/- 6) in the LAI assay if briefly preincubated with IgG obtained from LAI-position RA subjects. The LAI-positive response of PBL from RA patients was negated by preincubating the cells with sera obtained from LAI-nonreactive RA patients. Preincubation of reactive LAI-positive RA cells with urinary protein from 4 LAI-nonreactive RA patients blocked the response of reactive RA leukocytes in the LAI assay (42 +/- 2 to 14 +/- 2). In contrast, preincubation of reactive LAI-positive RA cells with urinary protein from LAI-reactive RA patients or patients with gout had no effect on subsequent LAI reactivity. This study suggests that LAI-nonreactive RA subjects have a 'rheumatoid neoantigen-like material' in their circulation which is excreted in their urine and is capable of being recognized by PBL of reactive, LAI-positive RA patients.


African Spilogona Schnabl: morphology of the male terminalia and description of a new species (Diptera: Muscidae).
Spilogona breviaristata sp. nov. from South Africa is described and the morphology of the male terminalia of seven African Spilogona Schnabl (Diptera, Muscidae) species are described and illustrated: Spilogona biguttata Emden, Spilogona fuscotriangulata Emden, Spilogona natalensis Zielke, Spilogona pertinisetodes Emden, Spilogona quasifasciata Emden, Spilogona semifasciata Emden and Spilogona spinipes (Bigot). The material studied is deposited in the Natural History Museum (BMNH), London, United Kingdom, and paratypes of the new species are also in the Museu Nacional, Universidade Federal do Rio de Janeiro (MNRJ), Rio de Janeiro, Brazil, and the Oxford University Museum of Natural History (OUMNH), Oxford, United Kingdom.


Mucosal Vaccination with Heterologous Viral Vectored Vaccine Targeting Subdominant SIV Accessory Antigens Strongly Inhibits Early Viral Replication.
Conventional HIV T cell vaccine strategies have not been successful in containing acute peak viremia, nor in providing long-term control. We immunized rhesus macaques intramuscularly and rectally using a heterologous adenovirus vectored SIV vaccine regimen encoding normally weakly immunogenic tat, vif, rev and vpr antigens fused to the MHC class II associated invariant chain. Immunizations induced broad T cell responses in all vaccinees. Following up to 10 repeated low-dose intrarectal challenges, vaccinees suppressed early viral replication (P=0.01) and prevented the peak viremia in 5/6 animals. Despite consistently undetectable viremia in 2 out of 6 vaccinees, all animals showed evidence of infection induced immune responses indicating that infection had taken place. Vaccinees, with and without detectable viremia better preserved their rectal CD4+ T cell population and had reduced immune hyperactivation as measured by naïve T cell depletion, Ki-67 and PD-1 expression on T cells. These results indicate that vaccination towards SIV accessory antigens vaccine can provide a level of acute control of SIV replication with a suggestion of beneficial immunological consequences in infected animals of unknown long-term significance. In conclusion, our studies demonstrate that a vaccine encoding subdominant antigens not normally associated with virus control can exert a significant impact on acute peak viremia.


Role of human cyclooxygenase-2 in the bioactivation of dapsone and sulfamethoxazole.
Sulfamethoxazole (SMX) and dapsone (4,4'-diaminodiphenylsulfone, DDS) are believed to mediate their adverse effects subsequent to bioactivation to their respective arylhydroxylamine and arylnitroso metabolites, resulting in covalent adduct formation with intracellular proteins. Various bioactivating enzymes, such as cytochromes P450 and myeloperoxidase, have been shown to be capable of catalyzing the N-oxidation of these compounds. We assessed the role of human cyclooxygenase-2 (COX-2) in the metabolism and subsequent adduct formation of DDS and SMX using recombinant human COX-2. Using an adduct-specific enzyme-linked immunosorbent assay, we found that the complete enzyme system gave rise to covalent adducts. However, the nonspecific COX inhibitor indomethacin did not reduce the amount of covalent adduct formed. Formation of the arylhydroxylamine metabolites was demonstrated via high performance liquid chromatography coupled with UV absorption. Metabolite formation was found to be secondary to the H2O2 in the incubation mixture and was not enzyme-mediated. Hence, COX-2 does not play a direct role in the bioactivation of these parent drugs to their arylhydroxylamine metabolites.


Chemical analysis and scanning electron microscopy of acquired pellicle formed in vivo on stannous fluoride treated enamel.
Stannous fluoride (SnF2) has been shown to be an effective caries preventive agent. After topical treatment of enamel surfaces, two reaction products have been demonstrated to precipitate on the surfaces, a larger type of globules, probably a calcium fluoride like product, and a smaller type of globules, probably a tin phosphate. The aim of the present study was to examine the amino acid composition and the formation of the acquired pellicle on SnF2-treated enamel in vivo. The chemical composition was examined by amino acid analysis of pellicle material collected in vivo from SnF2-treated enamel surfaces. Pellicle formation was examined by scanning electron microscopy on SnF2-treated enamel fragments carried in the mouth for 2 h. The results showed that pellicle material was formed in abundant amounts and covered the globular surfaces following the SnF2 treatment. The chemical analyses showed amino acid profiles with high content of acidic and neutral amino acids. The profiles were different from known amino acid profiles obtained from analyses of pellicle material collected from untreated enamel surfaces.


Effects of highly ordered TiO2 nanotube substrates on the nucleation of Cu electrodeposits.
We investigated the effects of TiO2 nanotube substrates on the nucleation density of Cu during electrodeposition in a solution of CuSO4 and H2SO4 at 50 degrees C compared with those of pure Ti and micro-porous TiO2 substrates. During electrodeposition, the density of Cu nuclei on the TiO2 nanotube substrate increased and the average size of Cu nuclei decreased with increasing anodizing voltage and time for the synthesis of the substrate. In addition, the nucleation density of Cu electrodeposits on the highly ordered TiO2 nanotube substrate was much higher than that on pure Ti and micro-porous TiO2 substrates.


Reflection-mode multifocal optical-resolution photoacoustic microscopy.
Compared with single-focus optical-resolution photoacoustic microscopy (OR-PAM), multifocal OR-PAM utilizes both multifocal optical illumination and an ultrasonic array transducer, significantly increasing the imaging speed. A reflection-mode multifocal OR-PAM system based on a microlens array that provides multiple foci as well as an ultrasonic array transducer that receives the excited photoacoustic waves from all foci simultaneously is presented. Using a customized microprism to reflect the incident laser beam to the microlens array, the multiple optical foci are aligned confocally with the focal zone of the ultrasonic array transducer. Experiments show the reflection-mode multifocal OR-PAM is capable of imaging microvessels in vivo, and it can image a 6×5×2.5 mm³ volume at 16 μm lateral resolution in ∼2.5 min, which was limited by the signal multiplexing ratio and laser pulse repetition rate.


[Biological cycle of the nematode Rictulariidae: Pterygodermatites (Multipectines) affinis (Jägerskiold, 1904) (author's transl)].
Infestant larva of the Nematode Rictulariidae: Pterygodermatities affinis (Jägerskiold, 1904) are localized in Pimelia: Tachyderma hispida Forskael of Algeria. Through the infestation of a young cat, embryonated eggs are collected after 38 days of development altogether with a female parasite. The experimental cycle of this rictulariid nematode in Locusta migratoria is characterized by the shortness of its development and the similarity in growth during the first and second larval stages. The cellular reaction of the Insect to the larva P. affinis is weaker in the Pimelia than in the Grasshopper, used as experimental host.


A case of tardive dyskinesia caused by metoclopramide.
Abnormal involuntary movements appeared in the mouth, tongue, neck and abdomen of a 64-year-old male patient after he took metoclopramide for gastrointestinal disorder in a regimen of 30 mg per day for a total of about 260 days. The symptoms exacerbated to a maximum in a month. When the metoclopramide administration was discontinued, the abnormal movements gradually improved to a considerable extent. Attention to the possible induction of specific tardive dyskinesia is called for in the use of this drug.


Nutrient intake and serum lipids in urban and rural Puerto Rican men.
Dietary intake information was collected on 2,426 rural and 5,828 urban men free of coronary heart disease in the Pureto Rico Heart Health Program. Serum cholesterol and triglyceride, body weights, and related parameters were measured during a detailed cardiovascular examination. There were statistically significant differences between rural and urban values with respect to lipids, relative weight, and intake of many nutrients. Rural subjects had lower lipids and relative weights. Serum cholesterol was associated with relative weight in both rural and urban areas. In the urban areas, serum cholesterol was associated with intake of total fat, saturated fatty acids, polyunsaturated fatty acids, total carbohydrate, and starch (all measured as percent of calories). It was also related to percent carbohydrate from starch and dietary cholesterol intake, even after relative weight was taken into account. The associations were of a low order, although statistically significant. The association of fasting triglycerides with some of the carbohydrate variables was statistically significant after relative weight was taken into account in the urban area. Overall, diet and relative weight can account for at most 6% of the variability in serum cholesterol observed, with at most 2.5% of the variability due diet alone.


[Team approach of Crohn's disease therapy].
Under the current conditions that etiologies of Crohn's Disease (CD) are not detected and then no radical therapy existed, it is necessary that the CD patients can be medically managed by not only medical doctors but also other co-medical staffs such as nurses, dietitians, social workers and ET (Enterostomal Therapist) nurses for their cure & care therapy. There are some different therapeutic steps; 1) Nutritional Therapy, 2) Drug Therapy, 3) Surgical Therapy and 4) Psychological (Mental) Support. In Japan, Nutritional Therapy headed by Elemental Enteral Nutrition is recommended as for 1st choice standard therapy for CD patients, who are mainly very young and are required to have themselves acceptable for long-term therapeutic controls. Thus, the medical support team shall be considerable enough to each patient's life back ground in order to give them the whole-mentally therapeutic approach.


hTERT C250T promoter mutation and telomere length as a molecular markers of cancer progression in patients with head and neck cancer.
Squamous cell carcinoma of the head and neck (HNSCC) is the sixth leading cause of cancer worldwide, representing over half a million incidents every year. Cancer cells, including HNSCC, are characterized by increased telomerase activity. This enzymatic complex is active in ~90% of all cancer types and is responsible for the lengthening of telomeres. Highly recurrent point mutations in the human telomerase reverse transcriptase (hTERT) promoter have recently been reported in a number of human neoplasms. The aim of the present study was to analyze the prevalence of the hTERT promoter C250T mutation and telomere length in the blood leukocytes of 61 patients with HNSCC and 49 healthy individuals. Quantitative polymerase chain reaction identified the hTERT promoter mutation in 36% of patients with HNSCC. To the best of our knowledge this is first report indicating the presence of shorter telomeres in early stage tumors. In addition, the results suggest that the C250T hTERT promoter mutation and telomere length assessment may serve as important molecular markers of HNSCC progression.