The establishment and validation of novel therapeutic targets to retard progression of chronic kidney disease.
The focus of this article is to define goals and resulting action plans that can be collectively embraced by interested stakeholders to facilitate new therapeutic approaches to mitigate chronic kidney disease progression. The specific goals include identifying druggable targets, increasing the capacity for preclinical and early clinical development, broadening the availability of new therapeutic approaches, and increasing investment in the development of new therapies to limit chronic kidney disease. Key deliverables include the establishment of new regional, national, and global consortia; development of clinical trial networks; and creation of programs to support the temporary mutual movement of scientists between academia and the biotechnology and pharmaceutical sector. Other deliverables include cataloging and maintaining up-to-date records to collate progress in renal research and development, inventorying the capacity of research and clinical networks, and describing methods to ensure novel drug development.


Differential gene expression profiles of human monocyte-derived antigen presenting cells in response to Penicillium marneffei: roles of DC-SIGN (CD209) in fungal cell uptake.
DNA microarray technology was used to determine the gene expression profile of human monocyte-derived macrophages (hMDMs) after stimulation by Penicillium marneffei yeast. The expression levels of 175 macrophage genes were found to be altered by a minimum of two-fold in magnitude following 4 hours of P. marneffei exposure. Among those, 41 genes were upregulated in activated hMDMs while 134 genes were downregulated. Real-time PCR and RT-PCR were performed to further examine gene expression associated with the inflammatory response. Increased levels of TNF-alpha and IL-1 beta gene expression in both hMDMs and human monocyte-derived dendritic cells (hMoDCs) were observed after stimulation by P. marneffei yeast. Furthermore, the genes encoding T-bet, IL-6 and ICAM-1 were also upregulated in hMDMs. Functional analysis of the adhesion of P. marneffei to dendritic cell-specific intercellular adhesion molecule-3 grabbing non-integrin (DC-SIGN, CD209) was performed in hMoDCs since the microarray data revealed an increased expression of DC-SIGN in activated hMDMs. We found that DC-SIGN-Fc bound preferentially to P. marneffei yeast rather than to conidia. Moreover, an anti-DC-SIGN monoclonal antibody inhibited the binding of P. marneffei yeast to hMoDCs, but did not inhibit endocytosis of P. marneffei yeast. The mannose receptor, on the other hand, was important in both adhesion and phagocytosis. These results suggest that P. marneffei may exploit DC-SIGN as a receptor to facilitate the systemic spread of infection. Taken together, our study demonstrates the usefulness of microarray technology in generating valuable expression data to permit conventional immunologic investigations of host-fungal interactions.


Psychometric Aspects of the Depressive Experiences Questionnaire: Implications for Clinical Assessment and Research.
The Depressive Experiences Questionnaire (DEQ) is a self-report measure that assesses self-criticism and dependency, 2 personality traits that confer vulnerability to depression (Blatt, 2004 ). Over several decades, different, shortened versions of the DEQ have been constructed to offer an alternative to the complex scoring procedure of the original DEQ. This study explores the factor structure as well as the construct and convergent validity of the DEQ by comparing a clinical and nonclinical sample. We also compared the original DEQ with 5 shortened versions. There were 621 participants (358 university students and 263 outpatients). Fit indexes for models of the original DEQ did not meet minimum fit criteria. Moreover, the only versions with satisfactory fit were the Theoretical Depressive Experiences Questionnaire-21 (TDEQ-21) and the Theoretical Depressive Experiences Questionnaire-12 (TDEQ-12), which also showed acceptable construct and convergent validity. Finally, the diagnostic and clinical applicability of the DEQ is discussed.


The impact of BMI on non-malignant respiratory symptoms and lung function in arsenic exposed adults of Northern Chile.
Elevated body mass index (BMI) and arsenic are both associated with cancer and with non-malignant lung disease. Using a unique exposure situation in Northern Chile with data on lifetime arsenic exposure, we previously identified the first evidence of an interaction between arsenic and BMI for the development of lung cancer. We examined whether there was an interaction between arsenic and BMI for the development of non-malignant lung disease. Data on lifetime arsenic exposure, respiratory symptoms, spirometry, BMI, and smoking were collected from 751 participants from cities in Northern Chile with varying levels of arsenic water concentrations. Spirometry values and respiratory symptoms were compared across subjects in different categories of arsenic exposure and BMI. Adults with both a BMI above the 90th percentile (>33.9kg/m2) and arsenic water concentrations ≥11µg/L exhibited high odds ratios (ORs) for cough (OR = 10.7, 95% confidence interval (CI): 3.03, 50.1), shortness of breath (OR = 14.2, 95% CI: 4.79, 52.4), wheeze (OR = 14.4, 95% CI: 4.80, 53.7), and the combined presence of any respiratory symptom (OR = 9.82, 95% CI: 4.22, 24.5). In subjects with lower BMIs, respiratory symptom ORs for arsenic water concentrations ≥11µg/L were markedly lower. In never-smokers, reductions in forced vital capacity associated with arsenic increased as BMI increased. Analysis of the FEV1/FVC ratio in never-smokers significantly increased as BMI and arsenic concentrations increased. Similar trends were not observed for FEV1 alone or in ever-smokers. This study provides preliminary evidence that BMI may increase the risk for arsenic-related non-malignant respiratory disease.


Wear simulation of ultra-high molecular weight polyethylene hip implants by incorporating the effects of cross-shear and contact pressure.
The effect of multi-directional cross-shear (CS) motion and contact pressure on ultra-high molecular weight polyethylene (UHMWPE) wear was investigated in this study, based on an integrated experimental and computational approach. The wear factor as a function of CS was determined experimentally from a multi-directional pin-on-plate wear tester under a nominal contact pressure of 1 MPa. A computational wear model was developed which included the effects of CS as well as the load and sliding distance imposed on the hip joint employing a UHMWPE cup against a metallic femoral head under both gait and Leeds ProSim hip joint simulator conditions. The CS ratios were quantified over the articular surface of the UHMWPE cup and the CS-dependent wear factors derived from multi-directional pin-on-plate studies were applied in the computational wear model. Outputs from the computational wear model were validated independently against an experimental hip simulator study. Comparisons of linear and volumetric wear were made between the computational wear model and the hip simulator testing for a nominal conventional (0 MRad) UHMWPE cup of 28mm diameter and a highly cross-linked (10 MRad) UHMWPE cup. The difference between the computed and experimental volumetric wear was approximately 30 per cent for the 0 MRad UHMWPE, although the worn areas between the prediction and the measurement were similar. For the 10 MRad UHMWPE, the discrepancy was reduced to 16 per cent. In both cases, the computational model predicted a lower wear rate than the experimental simulator testing. The effect of using alternative wear factors under a different nominal contact pressure of 3MPa was also considered. The input wear factor to the computational model, derived from a constant loaded pin-on-plate test configuration, may underestimate the dynamic effect due to the variation in the load in the hip joint simulator.


The optimal measure of microvascular function with velocity time integral for cardiovascular risk prediction.
Recent evidence suggests that microvascular function may be important in cardiovascular risk prediction. One measure of microvascular function is hyperaemic velocity time integral (VTI). We assessed whether the VTI of more than one beat of reactive hyperaemia would provide a stronger correlate to traditional cardiovascular risk factors using a subset of subjects from the Firefighters and Their Endothelium (FATE) study. Vascular function was assessed by measurement of hyperaemic blood velocity with high-resolution ultrasound of the brachial artery. We evaluated three measures in the current analysis: the VTI of the first beat, average VTI of 10 beats, and maximum VTI of 10 beats post-cuff release. A total of 399 male subjects (45.5 ± 10 years) were included in this analysis. Univariate correlations between the three end points and cardiovascular risk factors were calculated, and multivariable regression models constructed. Intra-observer variability was approximately equal for all VTI end points (coefficient of variation: first = 1.6%, average = 1.4%, maximum = 1.4%). Univariate correlations between VTI and cardiovascular risk factors were similar across all three end points. In multivariable analyses, there were no differences in the relationships between cardiovascular risk factors and the various VTI end points (R(2) from 0.090 to 0.102). Age, systolic blood pressure, and BMI were predictors of the three VTI end points (p < 0.05). In conclusion, the first beat of reactive hyperaemia remains the suitable measure of microvascular function.


Introduction of the videoscope in neurosurgery.
This article demonstrates that a newly developed videoscope for use in neurosurgery is effective for patients with lesions in their ventricles, cisterns, and intraparenchyma. Neuroendoscopy was performed on 13 patients using a videoscope system with narrow-band imaging technology. The results showed that the new videoscope was far superior to conventional fiberscopes in terms of visual quality and ease of manipulation. Observation of subependymal blood vessels, which had been difficult with fiberscopes, was possible with the videoscope. Furthermore, the videoscope was capable of producing enhanced images of capillary blood vessels under the ependymal layer space if used in conjunction with narrow-band imaging technology. This videoscope will be effective in avoiding capillary vessel injuries during neuroendoscopic procedures. It will also contribute to the future development of neuroendoscopy.


A molecular tweezer for lysine and arginine.
Lysine and arginine play a key role in numerous biological recognition processes controlling, inter alia, gene regulation, glycoprotein targeting and vesicle transport. They are also found in signaling peptide sequences responsible, e.g. for bacterial cell wall biosynthesis, Alzheimer peptide aggregation and skin regeneration. Almost none of all artificial receptor structures reported to date are selective and efficient for lysine residues in peptides or proteins. An artificial molecular tweezer is introduced which displays an exceptionally high affinity for lysine (K(a) approximately 5000 in neutral phosphate buffer). It features an electron-rich torus-shaped cavity adorned with two peripheral anionic phosphonate groups. Exquisite selectivity for arginine and lysine is achieved by threading the whole amino acid side chain through the cavity and subsequent locking by formation of a phosphonate-ammonium/guanidinium salt bridge. This pseudorotaxane-like geometry is also formed in small basic signaling peptides, which can be bound with unprecedented affinity in buffered aqueous solution. NMR titrations, NOESY and VT experiments as well as ITC measurements and Monte Carlo simulations unanimously point to an enthalpy-driven process utilizing a combination of van der Waals interactions and substantial electrostatic contributions for a conformational lock. Since DMSO and acetonitrile compete with the amino acid guest inside the cavity, a simple change in the cosolvent composition renders the whole complexation process reversible.


Draft Genome Sequence of Lactobacillus gorillae Strain KZ01T, Isolated from a Western Lowland Gorilla.
Here, we report the draft genome sequence of Lactobacillus gorillae strain KZ01(T) isolated from a western lowland gorilla (Gorilla gorilla gorilla). This genome sequence will be helpful for the comparative genomics between human and nonhuman primate-associated Lactobacillus.


Tissue-specific differences in activation of atypical protein kinase C and protein kinase B in muscle, liver, and adipocytes of insulin receptor substrate-1 knockout mice.
Insulin receptor substrates (IRSs) 1 and 2 are postulated to control the activation of phosphatidylinositol 3-kinase (PI3K)-dependent signaling factors, namely, atypical protein kinase C (aPKC) and protein kinase B (PKB)/Akt, which mediate metabolic effects of insulin. However, it is uncertain whether aPKC and PKB are activated together or differentially in response to IRS-1 and IRS-2 activation in insulin-sensitive tissues. Presently, we examined insulin activation of aPKC and PKB in vastus lateralis muscle, adipocytes, and liver in wild-type and IRS-1 knockout mice, and observed striking tissue-specific differences. In muscle of IRS-1 knockout mice, the activation of both aPKC and PKB was markedly diminished. In marked contrast, only aPKC activation was diminished in adipocytes, and only PKB activation was diminished in liver. These results suggest that IRS-1 is required for: 1) activation of both aPKC and PKB in muscle; 2) aPKC, but not PKB, activation in adipocytes; and 3) PKB, but not aPKC, activation in liver. Presumably, IRS-2 or other PI3K activators account for the normal activation of aPKC in liver and PKB in adipocytes of IRS-1 knockout mice. These complexities in aPKC and PKB activation may be relevant to metabolic abnormalities seen in tissues in which IRS-1 or IRS-2 is specifically or predominantly down-regulated.


DGKZ Acts as a Potential Oncogene in Osteosarcoma Proliferation Through Its Possible Interaction With ERK1/2 and MYC Pathway.
Osteosarcoma (OS) is one of the most common primary bone tumors in children and young adults. The majority of osteosarcoma patients have limited alternative therapeutic options and metastatic patients generally have a poor prognosis. Thus, it is important to explore novel effective therapeutic targets in the treatment of osteosarcoma. Diacylglycerol kinase zeta (DGKZ) is a recently identified gene potentially associated with certain human carcinogenesis. However, the role of DGKZ in proliferation of osteosarcoma is still unclear. In this study, DGKZ's expression was firstly investigated in OS tumor samples and correlated with poor outcome in OS patients. Silence of DGKZ by shRNA hampered osteosarcoma cell growth and promoted cell apoptosis in vitro. In vivo, DGKZ's knockout also suppressed xenograft tumor proliferation as determined by bioluminescence imaging and weight/volume measurements. Meanwhile, Affymetrix GeneChip and Ingenuity Pathway Analysis (IPA) revealed that DGKZ knockdown resulted in a decreased activity of MYC pathway, and several target genes expression in MYC pathway were altered, including CCND1, CDKN2B, CDK6, PCNA, and EGR1. Furthermore, immunoprecipitation coupled with mass spectrometry (IP-MS) analysis was used to identify proteins that interacted with DGKZ in OS cells and revealed ERK1/2, a key MYC-interactor, to associate with DGKZ. Together, our study demonstrated that DGKZ might act as an oncogene in osteosarcoma via its possible interaction with ERK1/2 and MYC pathway.


Effect of Rubus coreanus extracts on diabetic osteoporosis by simultaneous regulation of osteoblasts and osteoclasts.
Diabetes mellitus has been known to be associated with a high risk of osteoporosis. Rubus coreanus Miquel, a traditional Asian herbal medicine, has various uses, such as antiobesity and antiosteoporosis treatment, among others. We investigated the effect of R. coreanus extracts on diabetic osteoporosis. Rats were not treated, or treated with streptozotocin or R. coreanus, or ovariectomized, in various combinations. After 6 weeks of treatment, the rats were killed, and serum biochemistry, histopathology, immunohistochemistry, and semiquantitative reverse transcription polymerase chain reaction were performed. In addition, in vitro studies were performed in MC3T3-E1 and RAW 264.7 cells. Rats treated using R. coreanus showed significant improvement in trabecular bone histopathology. Increased expression of osteocalcin was observed in rats treated with streptozotocin and R. coreanus, whether ovariectomized or not. In addition, the expression levels of cannabinoid receptors 1 and 2 and receptor activator for nuclear factor κβ ligand were increased in rats that were ovariectomized and treated with streptozotocin and R. coreanus but decreased in those treated with streptozotocin and R. coreanus alone. These results indicate that the antiosteoporotic effect of R. coreanus in postmenopausal diabetic osteoporosis is attributable to the cannabinoid receptor-dependent maximal up-regulation of osteoblastogenesis. The present study shows that R. coreanus may rescue diabetic osteoporotic bone loss by simultaneous alteration of activation in osteoblasts and osteoclasts. Furthermore, these effects may be partially influenced by the up-regulation of the endocannabinoid system. In conclusion, dietary R. coreanus may be of use in improving the conditions of diabetic osteoporosis.


Scatter diagram analysis: a new technique for discriminating ventricular tachyarrhythmias.
With the increasing flexibility allowed by implantable cardioverter defibrillators that use tiered therapy, it is important to match the therapy with the arrhythmia. In this article we present scatter diagram analysis, a new computationally efficient two-channel algorithm for distinguishing monomorphic ventricular tachycardia (VT) from polymorphic ventricular tachycardia and ventricular fibrillation (VF). Scatter diagram analysis plots the amplitude from one channel versus the amplitude from another channel on a graph with a 15 x 15 grid. The fraction (percentage) of the 225 grid blocks occupied by at least one sample point is then determined. We found that monomorphic VT traces nearly the same path in space and occupies a smaller percentage of the graph than a nonregular rhythm such as polymorphic VT or VF. Scatter diagram analysis was tested on 27 patients undergoing intraoperative implantable cardioverter defibrillator testing. Passages of 4.096 seconds were obtained from rate (bipolar epicardial) and morphology (patch) leads, and digitized at 125 Hz. Scatter diagram analysis distinguished 13 episodes of monomorphic VT (28.6% +/- 4.0%) from 27 episodes of polymorphic VT or VF (48.0% +/- 8.2%) with P < 0.0005. There was overlap in only one monomorphic VT episode and one polymorphic VT or VF episode.


Cancer-associated fibroblast-derived Lumican promotes gastric cancer progression via the integrin β1-FAK signaling pathway.
Gastric cancer (GC) is one of the most frequent malignant tumors worldwide and is associated with high invasiveness, high metastasis and poor prognosis. Cancer-associated fibroblasts (CAFs), residing around tumor cells in tumor stroma, are important modifiers of tumor initiation and progression. However, the molecular mechanisms by which CAF's modulate tumor development have yet not to be characterized in GC. Here we performed tissue assay analyses identifying that Lumican, an extracellular matrix protein, is highly expressed in human gastric CAFs and its expression is positively associated with depth of invasion, lymph node metastasis, TNM stage and poor survival rate of GC. Functional studies revealed that integrin β1-FAK signaling pathways mediate the promoting effect of Lumican on GC cell proliferation, migration and invasion in vitro. In accordance with these observations, in GC cells co-cultured with CAFs in which Lumican had been knocked down, decreased gastric tumor growth and metastasis in vivo was apparent. In summary, CAF-derived Lumican contributes to tumorigenesis and metastasis of GC by activating the integrin β1-FAK signaling pathway.


Sociodemographic and Clinical Determinants of Time to Care-Seeking Among Febrile Children Under-Five in North-Central Nigeria.
Our study sought to determine the time parents of febrile children under the age of five took to seek competent medical care. We also looked at the possible sociodemographic/ clinical factors that influenced this presentation. Four hundred and nine under-fives presenting at the emergency unit with a history of fever in the last 48 hours along with their mothers were recruited over four months. Relevant sociodemographic information as well as symptoms and duration of illness were obtained. Multinomial regression analysis was performed to determine the predictors of early and late presentation. Over half (57%) of patients presented within 24 hours of onset of fever. The mean age of the children and mothers were 22±15 months and 30±5 years, respectively. High social class (odds ratio (OR) 6.5, 95% CI 1.6-26.4), Hausa ethnic group (OR 19.3, 95% CI 5.7-65.6), convulsions (OR 3.2, 95% CI 1.6-6.5) and appearance of other symptoms (OR 6.0, 95% CI 3.0-12.0) were significant predictors of early presentation. Secondary school education, belonging to another ethnic group, and non-resolution of fever were significant predictors of late presentation. The majority of febrile under-fives came to the hospital to seek competent medical care within the first 24 hours of illness. However, there is a need for more parental education on early hospital presentation for parents of low socioeconomic status and educational background.


Specific heat of molecular crystals from atomic mean square displacements with the Einstein, Debye, and Nernst-Lindemann models.
Analysis of atomic displacement parameters (ADPs) from multitemperature diffraction data provides mean-field molecular translation and libration frequencies. These quantities have been combined with molecular deformation frequencies calculated ab initio, e.g. by DFT methods, to calculate the specific heat Cv of molecular crystals of naphthalene, anthracene, and hexamethylenetetramine. If the difference Cp - Cv is represented by the Nernst-Lindemann relation, Cp curves from diffraction experiments and ab initio calculations agree well with those based on calorimetry. Agreement is better if the Debye rather than the Einstein model is chosen to represent the contribution of the translational vibrations. Compressibilities estimated from the differences Cp - Cv are 2-5 times higher than those obtained from compressibility measurements at 298 K and Grüneisen constants derived from the temperature dependence of ADPs.


Clinical utility of an immunoradiometric assay for parathyroid hormone (1-84) in primary hyperparathyroidism.
The reliable diagnosis of primary hyperparathyroidism depends on the measurement of PTH. The PTH assays in widespread use measure not only the hormone but also hormone fragments, thus limiting the clinical utility of the assays. A new immunoradiometric assay (IRMA) using an antigenic determinant at the extreme amino-terminal of the PTH molecule detects only full-length PTH (1-84). We compared three PTH assays and determined the presence of PTH (1-84) and PTH fragments in serum and parathyroid adenomas of patients with primary hyperparathyroidism. We studied 56 patients with primary hyperparathyroidism. PTH levels were increased in 63% using the midmolecule RIA; in 73% in the "intact" IRMA; and in 96% in the PTH (1-84)-IRMA. The PTH (1-84)-IRMA correlated with the other assays (midmolecule RIA R = +0.736; P < 0.0001; "intact"-IRMA R = +0.951; P < 0.0001) and indices of disease activity (serum calcium R = +0.511, P < 0.0001; alkaline phosphatase R = +0.489, P = 0.001; and radius bone density R = -0.366, P < 0.01). In 21 consecutive patients undergoing parathyroidectomy, 18 had parathyroid adenomas. Intact PTH was higher than PTH (1-84)-IRMA in both serum and glandular homogenates from these patients. Similar proportions of PTH (1-84) and hormone fragments were found in both adenomas [66 +/- 3% of "intact" PTH-reflected PTH (1-84) and sera (73 +/- 2% of "intact" PTH reflected PTH (1-84)]. We conclude that the PTH (1-84)-IRMA offers improved diagnostic sensitivity in patients with primary hyperparathyroidism than other currently available assays. This study also provides evidence that both PTH (1-84) and PTH fragments are produced in parathyroid adenomas and that peripheral metabolism of hormone and fragment does not alter the proportion of bioactive hormone.


Dueling biological and social contagions.
Numerous models explore how a wide variety of biological and social phenomena spread in social networks. However, these models implicitly assume that the spread of one phenomenon is not affected by the spread of another. Here, we develop a model of "dueling contagions", with a particular illustration of a situation where one is biological (influenza) and the other is social (flu vaccination). We apply the model to unique time series data collected during the 2009 H1N1 epidemic that includes information about vaccination, flu, and face-to-face social networks. The results show that well-connected individuals are more likely to get vaccinated, as are people who are exposed to friends who get vaccinated or are exposed to friends who get the flu. Our dueling contagion model suggests that other epidemiological models may be dramatically underestimating the R0 of contagions. It also suggests that the rate of vaccination contagion may be even more important than the biological contagion in determining the course of the disease. These results suggest that real world and online platforms that make it easier to see when friends have been vaccinated (personalized vaccination campaigns) and when they get the flu (personalized flu warnings) could have a large impact on reducing the severity of epidemics. They also suggest possible benefits from understanding the coevolution of many kinds of dueling contagions.


Difference in developmental toxicity among structurally similar N-phenylimide herbicides in rats and rabbits.
S-53482 is an N-phenylimide herbicide and shows a remarkable species difference in developmental toxicity between rats and rabbits. The herbicide produced embryolethality, teratogenicity (mainly ventricular septal defects and wavy ribs), and growth retardation in rats, but not in rabbits. Our objective in this study was to investigate differences in developmental toxicity among N-phenylimide compounds structurally similar to S-53482 to better characterize the developmental effects of S-53482 on rat and rabbit embryos as part of research investigation to elucidate a mechanism of rat developmental toxicity produced by S-53482. This paper is part of a series of studies to be published serially. S-23121 or S-23031, both of which are structurally similar to S-53482, was orally administered to rats and rabbits during fetal organogenesis. Fetuses were obtained by cesarian section and examined for external, visceral, and skeletal alterations. S-23121 produced the similar pattern of developmental toxicity such as embryolethality, teratogenicity, and growth retardation in rats. In contrast, S-23031 showed no developmental toxicity at 1500 mg/kg. Rabbits were insensitive to all study compounds. The difference in developmental toxicity in rats was striking among N-phenylimide herbicides. The mechanism of action of developmental toxicity by S-53482 should account for the compound-specific difference as well as species difference between rats and rabbits.


Tea drinking and cancer risk: epidemiologic evidence
Tea and tea compounds have been shown to inhibit carcinogenic processes in experimental animals, raising the possibility that tea drinking may lower cancer risk in humans. However, epidemiologic studies have produced inconsistent evidence on the relation between tea drinking and cancer risk. Ecological data show considerable international variation in tea consumption but relatively small differences in cancer rates. Results from case-control and cohort studies also are inconclusive. Nevertheless, high consumption of tea has been linked to a reduced risk of digestive tract cancers in a number of epidemiologic studies. A lack of detailed information on duration and amount of tea drinking, a narrow range of tea intake in some study populations, inadequate control for confounding, and potential biases in recall and reporting of tea drinking patterns in case-control studies may have contributed to the diverse findings. Further research is needed before definitive conclusions on tea's impact upon cancer risk in humans can be reached.


Nuclear localization of overexpressed glyceraldehyde-3-phosphate dehydrogenase in cultured cerebellar neurons undergoing apoptosis.
We recently reported that overexpression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC 188.8.131.52) is directly involved in cytosine arabinonucleoside (ara-C)- and low K+-induced neuronal death of cultured cerebellar granule cells. The former is entirely due to apoptosis, whereas the latter involves both apoptosis and necrosis. We examined the subcellular distribution of the overexpressed GAPDH occurring during apoptosis by using both subcellular fractionation and immunocytochemistry with a monoclonal antibody directed against this overexpressed protein. When immature cerebellar neurons were exposed to ara-C, an overexpression of GAPDH was observed, primarily in the nuclear fraction. In contrast, low K+ exposure of mature cerebellar neurons induced the overexpression of GAPDH not only in the nuclear fraction but also in the mitochondrial fraction. In both paradigms, no significant change of GAPDH levels occurred in the microsomal and cytosolic fractions. Moreover, pretreatment with GAPDH antisense oligonucleotide or classic apoptotic inhibitors clearly suppressed the accumulation of GAPDH protein in these subcellular loci. This discrete nuclear localization of GAPDH during apoptosis was supported further by immunoelectron microscopy. Quantitative assessment of GAPDH immunogold labeling revealed that a approximately 5-fold increase in the intensity of gold particles was observed within the nucleus of apoptotic cells. Thus, the current results raise the possibility that neuronal apoptosis may be triggered by GAPDH accumulation in the nucleus, resulting in perturbation of nuclear function and ultimate cell death.


Parallax method for direct measurement of membrane penetration depth utilizing fluorescence quenching by spin-labeled phospholipids.
This report describes a method suitable for determining the depth of a wide variety of fluorescent molecules embedded in membranes. The method involves determination of the parallax in the apparent location of fluorophores detected when quenching by phospholipids spin-labeled at two different depths is compared. By use of straightforward algebraic expressions, the method allows calculation of depth in angstroms. Furthermore, the analysis can be extended to quenching by energy-transfer acceptors or brominated probes under appropriate conditions. Application of the method to quenching of 7-nitro-2,1,3-benzoxadiazol-4-yl (NBD)-labeled lipids by spin-labeled lipids located at three different depths is demonstrated in model membranes. It is shown that the calculated depths of the NBD groups are self-consistent to the extent that they are the same no matter which two spin-labels have been used in a particular experiment. In addition, the calculated depth is independent of spin-label concentration in the membrane within +/- 1 A, ruling out major effects due to spin-label perturbation. The quenching experiments show that the location of the NBD group in head-group-labeled phosphatidylethanolamine is at the polar/hydrocarbon interface and that of an NBD label on the "tail" of cholesterol is deeply buried, as expected. Unexpectedly, NBD labels placed at the end of fatty acyl chains of phosphatidylcholines are also near the polar/hydrocarbon interface. Presumably, the polarity of the NBD group results in "looping" back to the surface of the NBD groups attached to flexible acyl chains.


Current status and prospect of treatments for recurrent hepatocellular carcinoma.
Owing to its heterogeneous and highly aggressive nature, hepatocellular carcinoma (HCC) has a high recurrence rate, which is a non-negligible problem despite the increasing number of available treatment options. Recent clinical trials have attempted to reduce the recurrence and develop innovative treatment options for patients with recurrent HCC. In the event of liver remnant recurrence, the currently available treatment options include repeat hepatectomy, salvage liver transplantation, tumor ablation, transcatheter arterial chemoembolization, stereotactic body radiotherapy, systemic therapies, and combination therapy. In this review, we summarize the strategies to reduce the recurrence of high-risk tumors and aggressive therapies for recurrent HCC. Additionally, we discuss methods to prevent HCC recurrence and prognostic models constructed based on predictors of recurrence to develop an appropriate surveillance program.


A Trial of the Effect of 2-Step Orthosis Treatment for Mallet Finger Fractures.
Many patients report poor therapeutic outcomes following mallet finger fracture surgery. A more reliable technique is urgently needed. To present a novel treatment for mallet finger fractures using a 2-step orthosis method. Prospective, observational study. Hospital. Patients with mallet finger fractures. The finger is fixed with splints for 6 weeks, including 3 weeks for the proximal interphalangeal joint in the flexion position and the distal interphalangeal joint in the hyperextension position (first splint) and 3 weeks for the distal interphalangeal joint in the hyperextension position (second splint). Up to week 8, the second splint was attached at night and during physical exertion. Crawford criteria, Abouna-Brown criteria, bone fusion, grip strength, Doyle classification, Ishiguro classification. Sufficient bone fusion was achieved 12 weeks after fixation; at which time, the range of motion with the distal interphalangeal joint flexed, and extended in the 3 patients was 50° and 0°, 70° and -3°, and 60° and 0°, respectively. The right and left hand grip strengths in the 3 patients were 58 and 55 kg, 62 and 58 kg, and 31 and 29 kg, respectively; there were no problems with respect to function or work. The first 2 patients could start sports again with partial return after 1 week and complete return after 12 weeks and 8 weeks, respectively. For the third patient, rehabilitation was complete after 16 weeks. Evaluation of the fracture sites based on the Crawford criteria showed the condition to be perfect, and evaluation based on the Abouna-Brown criteria showed success. This method provides satisfactory fixation and can prevent proximal interphalangeal joint contracture. Favorable long-term outcomes were confirmed in all patients, suggesting that this method may be effective for previously untreated mallet finger fractures with little displacement.


The impact of COVID-19 in an assisted living community.
The purpose of this study was to use routinely collected resident assessment data from a single site to evaluate the impact of COVID-19 restrictions on cognition, physical function and behavioral symptoms of residents with dementia. Specifically, it was hypothesized that controlling for age and sex, there would be a decline in cognition and function and an increase in behavioral symptoms at 12 months post implementation of COVID-19 restrictions. Twelve residents from a single memory care site with required Resident Assessments completed prior to and 12 month post initiation of the pandemic and associated quarantines were included. No significant change was noted in function or behavioral symptoms but there was a statistically significant decline in cognition over the 12 month period. Although this study did not support our hypothesis, the findings supported some prior research also noting little significant change among the majority of individuals over the course of the pandemic regardless of regulations.


Conserved redox-dependent DNA binding of ROXY glutaredoxins with TGA transcription factors.
The Arabidopsis thaliana CC-type glutaredoxin (GRX) ROXY1 and the bZIP TGA transcription factor (TF) PERIANTHIA (PAN) interact in the nucleus and together regulate petal development. The CC-type GRXs exist exclusively in land plants, and in contrast to the ubiquitously occurring CPYC and CGFS GRX classes, only the CC-type GRXs expanded strongly during land plant evolution. Phylogenetic analyses show that TGA TFs evolved before the CC-type GRXs in charophycean algae. MpROXY1/2 and MpTGA were isolated from the liverwort Marchantia polymorpha to analyze regulatory ROXY/TGA interactions in a basal land plant. Homologous and heterologous protein interaction studies demonstrate that nuclear ROXY/TGA interactions are conserved since the occurrence of CC-type GRXs in bryophytes and mediated by a conserved ROXY C-terminus. Redox EMSA analyses show a redox-sensitive binding of MpTGA to the cis-regulatory as-1-like element. Furthermore, we demonstrate that MpTGA binds together with MpROXY1/2 to this motif under reducing conditions, whereas this interaction is not observed under oxidizing conditions. Remarkably, heterologous complementation studies reveal a strongly conserved land plant ROXY activity, suggesting an ancestral role for CC-type GRXs in modulating the activities of TGA TFs. Super-resolution microscopy experiments detected a strong colocalization of ROXY1 with the active form of the RNA polymerase II in the nucleus. Together, these data shed new light on the function of ROXYs and TGA TFs and the evolution of redox-sensitive transcription regulation processes, which likely contributed to adapt land plants to novel terrestrial habitats.


Hormone replacement therapy is associated with improved survival in women with advanced heart failure.
We sought to determine whether hormone replacement therapy (HRT) is associated with an improved prognosis in women with advanced heart failure (HF) and systolic dysfunction. There are about two million postmenopausal women in the U.S. with HF. However, limited data are available to assess the effects of HRT on survival in this large group of patients. A retrospective analysis of women age 50 years and over entered into the Beta-Blocker Evaluation of Survival Trial (BEST) was conducted using Cox regression analysis comparing survival in HRT users and non-users after correcting for baseline variables known to predict survival in women with HF and systolic dysfunction. In 493 women age 50 years and older, HRT was associated with a significant reduction in mortality-21% mortality in HRT users and 34% in non-users (p = 0.025). Multivariate analysis demonstrated a hazard ratio for mortality of 0.6 (95% confidence interval = 0.36 to 0.97) (p = 0.039) for HRT users. The benefits of HRT were noted only in women with a nonischemic etiology of HF (n = 237). Hormone replacement therapy is associated with a marked improvement in survival in postmenopausal women with advanced HF. A prospective, randomized trial of HRT should be performed in this large group of patients.


Cytomegalovirus: epidemiology and infection control.
Cytomegalovirus (CMV) causes asymptomatic infection in most individuals but can produce devastating illness in immunocompromised hosts and in a small proportion of congenitally infected babies. New techniques in molecular biology have provided insights into the epidemiology and transmission of CMV. Children in day care, their parents, and sexually active individuals, especially homosexual men, are now known to be at particular risk for acquiring CMV. Recent studies show that the risk of CMV acquisition by health care workers is similar to the risk to the general public. Health care workers should be aware of the wide range of clinical manifestations, methods of laboratory diagnosis, and current limitations of treatment of CMV. Careful handwashing and avoidance of excretions and secretions are recommended to decrease the transmission of CMV in the hospital.


Evaluation of the uterine environment and embryos of prepubertal gilts.
A series of three experiments was conducted to test the functional status of the uterus and embryos in prepubertal gilts. In Exp. 1, gilts were induced to ovulate by treating with gonadotropins followed by hCG 72 or 96 h later, and were artificially inseminated 24 h after hCG. Five of the 10 gilts treated at 120 d of age, but none of the gilts treated at 100 of age, maintained pregnancies. We next tested the function of the uterine environment by transferring embryos from postpubertal females into gilts of various ages that had been induced to ovulate but not inseminated (Exp. 2). Pregnancy rate at d 50 of gestation was 44% (4/9) for 100-d-old recipients, 67% (2/3) for 140-d-old recipients, and 60% (3/5) for postpubertal recipients (P > 0.20). Therefore, uteri of 100-d-old gilts are able to maintain pregnancies with conceptuses from postpubertal gilts. In Exp. 3, embryos from 100-d-old and postpubertal gilts were transferred into postpubertal recipients. Uterine horns of recipients were surgically separated before transfer, and embryos from 100-d-old and post-pubertal females were transferred to opposite horns of some recipients (experimental). Other recipients received embryos from postpubertal females in both uterine horns (control). When examined on d 50 to 60 of gestation, three of five control gilts were pregnant and three of seven experimental gilts were pregnant (P > 0.50). In experimental recipients, the survival of embryos from 100-d-old gilts was 38% (8/21) compared to 57% (15/26) for embryos from postpubertal gilts (P > 0.30). Because all uterine horns of pregnant recipients contained fetuses, these results support the hypothesis that embryos from 100-d-old gilts are able to initiate and maintain pregnancies in the uteri of postpubertal gilts. Therefore, the uterine environment of 100-d-old gilts provides an environment that supports development of embryos produced by postpubertal gilts, and the embryos produced by 100-d-old gilts can survive and develop in the uteri of postpubertal gilts. It was only the combination of embryos and uteri of 100-d-old gilts that did not permit pregnancy to be maintained.


[Carcinoma in situ of the breast].
Ductal carcinoma in situ (DCIS) and lobular carcinoma in situ (LCIS) represent a proliferation of malignant epithelial cells within the ducts of the breast without invasion through the basement membrane. It is assumed that all invasive carcinomas of the breast are preceded by DCIS or LCIS. However, it is not known what proportion of DCIS or LCIS--if left untreated--will progress to invasive carcinoma. Standard treatment for DCIS is breast-conserving surgery with free margins and radiation therapy. LCIS is usually treated by observation alone.


The SCL gene product is regulated by and differentially regulates cytokine responses during myeloid leukemic cell differentiation.
Differentiation induction in murine M1 leukemia cells by interleukin 6 (IL-6), leukemia inhibitory factor (LIF), and oncostatin M (OSM) is postulated to occur via a common receptor chain, gp130. In this study, growth factor-induced differentiation of M1 cells was accompanied by a late and persistent decrease in levels of mRNA and protein for a helix-loop-helix transcription factor, the SCL gene product. To evaluate whether reduced SCL expression was instrumental in monocyte differentiation, an SCL cDNA expression vector was introduced into M1 cells to obtain cell lines in which overexpression of SCL mRNA and protein was enforced. This resulted in a reduction in cells differentiating in response to LIF and OSM but not in response to IL-6. Scatchard analysis indicated that both parental and SCL-transfected cell lines exhibited similar receptor numbers and receptor affinities for LIF, OSM, and IL-6, suggesting that the differential responsiveness was not due to selective receptor down-modulation. Thus, these data implicate SCL in monocytic differentiation and provide evidence for differential receptor signaling pathways despite utilization of a common gp130 subunit by all three receptors.


Plan delivery quality assurance for CyberKnife: Statistical process control analysis of 350 film-based patient-specific QAs.
Robotic radiosurgery requires plan delivery quality assurance (DQA) but there has never been a published comprehensive analysis of a patient-specific DQA process in a clinic. We proposed to evaluate 350 consecutive film-based patient-specific DQAs using statistical process control. We evaluated the performance of the process to propose achievable tolerance criteria for DQA validation and we sought to identify suboptimal DQA using control charts. DQAs were performed on a CyberKnife-M6 using Gafchromic-EBT3 films. The signal-to-dose conversion was performed using a multichannel-correction and a scanning protocol that combined measurement and calibration in a single scan. The DQA analysis comprised a gamma-index analysis at 3%/1.5mm and a separate evaluation of spatial and dosimetric accuracy of the plan delivery. Each parameter was plotted on a control chart and control limits were calculated. A capability index (Cpm) was calculated to evaluate the ability of the process to produce results within specifications. The analysis of capability showed that a gamma pass rate of 85% at 3%/1.5mm was highly achievable as acceptance criteria for DQA validation using a film-based protocol (Cpm>1.33). 3.4% of DQA were outside a control limit of 88% for gamma pass-rate. The analysis of the out-of-control DQA helped identify a dosimetric error in our institute for a specific treatment type. We have defined initial tolerance criteria for DQA validations. We have shown that the implementation of a film-based patient-specific DQA protocol with the use of control charts is an effective method to improve patient treatment safety on CyberKnife.


Theiler's virus infection of perforin-deficient mice.
Theiler's virus, a murine picornavirus, infects the central nervous systems of C57BL/6 mice and is cleared after approximately 10 days by a process which requires CD8+ cytotoxic T cells. We used perforin-deficient C57BL/6 mice to test the role of this protein in viral clearance. Perforin-deficient mice died from viral encephalomyelitis between days 12 and 18 postinoculation. They had high levels of viral RNA in their central nervous systems until the time of death. In contrast, viral RNA had disappeared by day 11 postinoculation in wild-type C57BL/6 mice. Cytotoxic T cells can kill infected cells by two main mechanisms: the secretion of the pore-forming protein perforin or the interaction of the Fas ligand with the apoptosis-inducing Fas molecule on the target cell. Our results demonstrate that clearance of Theiler's virus from the central nervous system in C57BL/6 mice is perforin dependent.


Live/real time three-dimensional transthoracic echocardiographic assessment of aortic dissection rupture into right ventricular outflow tract: a case report and review of the literature.
We present a case of chronic ascending thoracic aortic dissection with rupture into the right ventricular outflow tract (RVOT) diagnosed by two-dimensional transthoracic echocardiogram in which live/real time three-dimensional (3D) transthoracic echocardiogram provided incremental value by demonstrating: (a) a tortuous false lumen that encroached and ruptured into the RVOT, (b) exact location of the rupture site in relation to other surrounding structures in 3 dimensions (c) en face view of the rupture site facilitating assessment of its size and shape, and (d) localized compression of the main pulmonary artery (PA) by the false lumen. In addition, cropping of the 3D data set permitted visualization of the origin of the left main coronary in a rapid manner excluding involvement of this vessel with the dissection process. These findings have potential implications for surgical planning and were corroborated by a computed tomography angiogram. We summarize seven previously reported aortic dissection cases with rupture into the right ventricle.


Expression Analysis of GRHL3 and PHLDA3 in Head and Neck Squamous Cell Carcinoma.
Head and neck squamous cell carcinoma (HNSCC) includes a group of heterogeneous tumors with generally invasive behavior. The PI3K/AKT pathway plays an important role in the pathogenesis of HNSCC. In the current study, we investigated the expression of two negative feedback regulators of the PI3K pathway, namely PHLDA3 and GRHL3, in 45 paired samples of HNSCC and adjacent non-cancerous tissues (ANCTs). While expression of GRHL3 was down-regulated in tumoral tissues compared with ANCTs by the factor 4.21, PHLDA3 expression levels were up-regulated by 5.99-times. Gender-based analysis revealed a significant down-regulation of GRHL3 gene expression level in male patients compared with the control samples and significant up-regulation of PHLDA3 gene expression level in both sexes compared with the control samples. Differences in the expressions of both genes were significant in patients aged more than 60 years, but not in the younger patients. Expression of GRHL3 was only down-regulated in patients with positive smoking history. Expression of GRHL3 was decreased in grades 2 and 3 samples compared with controls. There was a significant increase in transcript levels of PHLDA3 in stages II and III HNSCC samples compared with the controls group. ROC curve analysis indicated that the expression level of PHLDA3 could be a promising marker for the diagnosis of HNSCC patients with a sensitivity and specificity of 0.666 and 0.688, respectively. In addition, sensitivity and specificity of GRHL3 were 0.755 and 0.577, respectively. The current study indicates dysregulation of regulators of PI3K pathway in HNSCC and their potential application as putative biomarkers for this cancer.


Reversal of Neuronal Atrophy: Role of Cellular Immunity in Neuroplasticity and Aging.
Emerging evidence indicates that neuroimmunological changes in the brain can modify intrinsic brain processes that are involved in regulating neuroplasticity. Increasing evidence suggests that in some forms of motor neuron injury, many neurons do not die, but reside in an atrophic state for an extended period of time. In mice, facial motor neurons in the brain undergo a protracted period of degeneration or atrophy following resection of their peripheral axons. Reinjuring the proximal nerve stump of the chronically resected facial nerve stimulates a robust reversal of motor neuron atrophy which results in marked increases in both the number and size of injured motor neurons in the facial motor nucleus. In this brief review, we describe research from our lab which indicates that the reversal of atrophy in this injury model is dependent on normal cellular immunity. The role of T cells in this unique form of neuroplasticity following injury and in brain aging, are discussed. The potential role of yet undiscover intrinsic actions of recombination activating genes in the brain are considered. Further research using the facial nerve reinjury model could identify molecular signals involved in neuroplasticity, and lead to new ways to stimulate neuroregenerative processes in neurotrauma and other forms of brain insult and disease.


The Standardisation of handoffs in a large academic paediatric emergency department using I-PASS.
Despite the American College of Emergency Physicians and American Academy of Pediatrics recommendations for standardised handoffs in the emergency department (ED), few EDs have an established tool. Our aim was to improve the quality of handoffs in the ED by establishing compliance with the I-PASS handoff tool. This is a quality improvement (QI) initiative to standardise handoffs in a large academic paediatric ED. Following review of the literature and focus groups with key stakeholders, I-PASS was selected and modified to fit departmental needs. Implementation throughPlan-Do-Study-Act cycles included the development of educational materials, reminders and real-time feedback. Required use of I-PASS during designated team sign-out began in June 2016. Compliance with the handoff tool and handoff deficiencies was measured through observations by faculty trained in I-PASS. As a balancing measure, time to complete handoff was monitored and compared with preintervention data. Compliance with I-PASS reached 80% within 6 months, 100% within 7 months and sustained at 100% during the remainder of the study period. The average percent of omissions of crucial information per handoff declined to 8.3%, which was a 53% decrease. Average percentage of tangential information and miscommunications per handoff did not show a decline. The average handoff took 20 min, which did not differ from the preintervention time. Survey results demonstrated a perceived improvement in patient safety through closed-loop communication, clear action lists and contingency planning and proper patient acuity identification. I-PASS is applicable in the ED and can be successfully implemented through QI methodology contributing to an overall culture of safety.


LTP saturation and spatial learning disruption: effects of task variables and saturation levels.
The prediction that "saturation" of LTP/LTE at hippocampal synapses should impair spatial learning was reinvestigated in the light of a more specific consideration of the theory of Hebbian associative networks, which predicts a nonlinear relationship between LTP "saturation" and memory impairment. This nonlinearity may explain the variable results of studies that have addressed the effects of LTP "saturation" on behavior. The extent of LTP "saturation" in fascia dentata produced by the standard chronic LTP stimulation protocol was assessed both electrophysiologically and through the use of an anatomical marker (activation of the immediate-early gene zif268). Both methods point to the conclusion that the standard protocols used to induce LTP do not "saturate" the process at any dorsoventral level, and leave the ventral half of the hippocampus virtually unaffected. LTP-inducing, bilateral perforant path stimulation led to a significant deficit in the reversal of a well-learned spatial response on the Barnes circular platform task as reported previously, yet in the same animals produced no deficit in learning the Morris water task (for which previous results have been conflicting). The behavioral deficit was not a consequence of any after-discharge in the hippocampal EEG. In contrast, administration of maximal electroconvulsive shock led to robust zif268 activation throughout the hippocampus, enhancement of synaptic responses, occlusion of LTP produced by discrete high-frequency stimulation, and spatial learning deficits in the water task. These data provide further support for the involvement of LTP-like synaptic enhancement in spatial learning.


Inter-patient image registration algorithms to disentangle regional dose bioeffects.
Radiation therapy (RT) technological advances call for a comprehensive reconsideration of the definition of dose features leading to radiation induced morbidity (RIM). In this context, the voxel-based approach (VBA) to dose distribution analysis in RT offers a radically new philosophy to evaluate local dose response patterns, as an alternative to dose-volume-histograms for identifying dose sensitive regions of normal tissue. The VBA relies on mapping patient dose distributions into a single reference case anatomy which serves as anchor for local dosimetric evaluations. The inter-patient elastic image registrations (EIRs) of the planning CTs provide the deformation fields necessary for the actual warp of dose distributions. In this study we assessed the impact of EIR on the VBA results in thoracic patients by identifying two state-of-the-art EIR algorithms (Demons and B-Spline). Our analysis demonstrated that both the EIR algorithms may be successfully used to highlight subregions with dose differences associated with RIM that substantially overlap. Furthermore, the inclusion for the first time of covariates within a dosimetric statistical model that faces the multiple comparison problem expands the potential of VBA, thus paving the way to a reliable voxel-based analysis of RIM in datasets with strong correlation of the outcome with non-dosimetric variables.


Seasonal survival and reversible state effects in a long-distance migratory shorebird.
Events during one stage of the annual cycle can reversibly affect an individual's condition and performance not only within that stage, but also in subsequent stages (i.e. reversible state effects). Despite strong conceptual links, however, few studies have been able to empirically link individual-level reversible state effects with larger-scale demographic processes. We studied both survival and potential reversible state effects in a long-distance migratory shorebird, the Hudsonian Godwit Limosa haemastica. Specifically, we estimated period-specific survival probabilities across the annual cycle and examined the extent to which an individual's body condition, foraging success and habitat quality during the nonbreeding season affected its subsequent survival and reproductive performance. Godwit survival rates were high throughout the annual cycle, but lowest during the breeding season, only slightly higher during southbound migration and highest during the stationary nonbreeding season. Our results indicate that overwintering godwits foraging in high-quality habitats had comparably better nutritional status and pre-migratory body condition, which in turn improved their return rates and the likelihood that their nests and chicks survived during the subsequent breeding season. Reversible state effects thus appeared to link events between nonbreeding and breeding seasons via an individual's condition, in turn affecting their survival and subsequent reproductive performance. Our study thus provides one of the few empirical demonstrations of theoretical predictions that reversible state effects have the potential to influence population dynamics.


Solvent-mediated molar conductivity of protic ionic liquids.
The molar conductivity, Λm, of protic ionic liquids (PILs) in molecular solvents is measured at 298.15 K. The decrease in the Λm values of PILs is observed with an increase in the concentration of PILs. The limiting molar conductivities, Λm(0), were obtained for each PIL in different molecular solvents using a least squares method. The Λm(0) data for PILs were correlated with the structural aspects of PILs and solvent properties. The polar protic solvents show poor ionic association as compared to the polar aprotic solvents, which is discussed on the basis of the hydrogen bond donating (HBD) ability of solvents and PILs. The alkyl chain substitution of anions plays a significant role in the ionic association of the PILs. The diffusion coefficient D(0) and the transport number t were determined, which were consistent with the Λm(0) values of PILs in water. The Λm(0) and D(0) values are dependent on the hydrodynamic radius of anions of these ionic liquids. The extent of ionic association for each PIL was discussed using temperature dependent Λm data for aqueous PIL systems in terms of the Walden plot.


Gastrointestinal tumor-related perihepatic fluorouracil encapsulated lesions and liver metastases: a diagnostic imaging study based on contrast-enhanced computed tomography and magnetic resonance imaging.
Perihepatic fluorouracil encapsulated lesions (FELs) can result in potentially confusing computed tomography (CT) and magnetic resonance imaging (MRI) features in postoperative examinations of gastrointestinal tumors. This retrospective study aimed to summarize the typical imaging features of FELs and determine the best imaging modality to distinguish FELs from liver metastases for junior residents. Patients with FELs who had undergone gastrointestinal tumor surgery in Tongji Hospital from January 2016 to June 2022 were evaluated. The imaging features of FELs were summarized by two senior radiologists. Contrast-enhanced CT (CECT) was used as the primary follow-up tool for postoperative gastrointestinal tumor patients. Patients with FELs and available CECT and MRI examinations were matched with patients with liver metastases based on gender and age and presented in chronological order in a 2:1 ratio. Different imaging modality combinations were used for further evaluation, including a CECT group (modality Ⅰ), CECT and nonenhanced MRI group (modality Ⅱ) and CECT with all MRI sequences group (modality Ⅲ). Subsequently, two junior residents blindly evaluated three groups following a 4-week interval based on a 5-point scale (1= definite benign lesion, 2= probable benign lesion, 3= indeterminate, 4= probable liver metastasis, 5= definite liver metastasis). Imaging features of 33 patients with 36 FELs were analyzed. CECT and dynamic contrast-enhanced MRI (DCE-MRI) showed no enhancement in most lesions. Additionally, 20 patients with FELs meeting the requirements were matched with 40 patients with liver metastases. The highest sensitivity, specificity, and consistency for identifying liver metastases were achieved using a combination of CECT and MRI encompassing all sequences yielded, including modality Ⅰ (reader 1: 72.0% and 17.4%; reader 2: 62.0% and 17.4%; kappa value 0.295), modality Ⅱ (reader 1: 88.0% and 8.7%; reader 2: 92.0% and 34.8%; kappa value 0.259), and modality Ⅲ (reader 1: 98.0% and 34.8%; reader 2: 92.0% and 39.1%; kappa value 0.680). FELs are typically non-enhancing lesions. In our study, two junior residents could best distinguish FELs from liver metastases using CECT with all MRI sequences.


Between Scylla and Charybdis: risks of early therapeutic anticoagulation for venous thromboembolism after acute intracranial hemorrhage.
To assess the risk of hematoma expansion in patients with acute intracranial hemorrhage (ICH) requiring therapeutic anticoagulation for the treatment of venous thromboembolism. We retrospectively reviewed all patients at our institution between 2014 and 2019 who were therapeutically anticoagulated for venous thromboembolism within 4 weeks after ICH. We included subtypes of traumatic ICH and spontaneous intraparenchymal hemorrhage. Our main outcome was the incidence of hematoma expansion within 14 days from initiating therapeutic anticoagulation. Hematoma expansion was defined as (1) radiographically proven expansion leading to cessation of therapeutic anticoagulation or (2) death due to hematoma expansion. Secondary outcomes included mortality due to hematoma expansion and characteristics associated with hematoma expansion. Fifty patients met inclusion criteria (mean age: 54 years, 80% male, 76% Caucasian); 24% had undergone a neurosurgical procedure prior to therapeutic anticoagulation. Median time from ICH to therapeutic anticoagulation initiation was 9.5 days (IQR 4-17), 40% received therapeutic anticoagulation in <7 days after ICH. Six patients (12%) developed hematoma expansion, of whom two (4%) died. While not statistically significant, patients with hematoma expansion tended to be older (57.8 vs. 53.5 years), were anticoagulated sooner (4 vs. 10 days), presented with lower GCS (50% vs. 39% with GCS <8), higher hematoma volume (50% vs. 42% >30 cc), and higher SDH diameter (16 mm vs. 8.35 mm). There was a trend towards greater risk of hematoma expansion for patients undergoing endoscopic ICH evacuation (16% vs. 2%, p = 0.09); patients with hematoma expansion were more likely to present with hydrocephalus (67% vs. 16%, p = 0.02). Our study is among the first to explore characteristics associated with hematoma expansion in patients undergoing therapeutic anticoagulation after acute ICH. Larger studies in different ICH subtypes are needed to identify determinants of hematoma expansion in this high-acuity population.


Risk perceptions and family history of lung cancer: differences by smoking status.
Individuals with a family history of lung cancer have a two- to threefold increased risk for developing this disease. Family history information may be useful in lung cancer prevention and control approaches, but research is needed regarding how individuals interpret this information. This study examined associations between lung cancer family history and individuals' risk perceptions, based on smoking status. Data were analyzed from 5,105 U.S. adult respondents to the 2005 Health Information National Trends Survey, which was conducted with a nationally representative sample. In multivariate models, family history of lung cancer was positively associated with absolute and relative risk perceptions among all respondents (β=0.60, 95% CI=0.33-0.87 and β=0.17, 95% CI=0.04-0.31, respectively) and among never smokers (β=0.40, 95% CI=0.14-0.67 and β=0.14, 95% CI=0.01-0.27, respectively). However, these associations were not significant for current and former smokers. While perceived risk was associated with family history of lung cancer among never smokers, this was not true for other smoking status subgroups. Therefore, former and current smokers might not respond as intended to cancer prevention or cessation messages tailored based on family history. The results suggest directions for future research into how to best integrate family history information into prevention and control efforts.


CD4+CD25+ regulatory T cell therapy for the induction of donor-specific clinical transplantation tolerance.
Naturally occurring CD4+CD25+ regulatory T cells (Tregs) have been shown to play a key role in the control of autoimmunity. Interestingly, they are also capable of mediating transplantation tolerance and they can have indirect allospecificity for donor antigens. An increasing body of evidence in experimental studies has indicated that adoptive transfer of in vitro expanded CD4+CD25+ Tregs with indirect antidonor allospecificity can induce long-term donor-specific transplantation tolerance. Thus, adoptive cell therapy using patient-specific CD4+CD25+ Tregs as individualised medicine to promote clinical transplantation tolerance is promising.


Comparative mucomic analysis of three functionally distinct Cornu aspersum Secretions.
Every animal secretes mucus, placing them among the most diverse biological materials. Mucus hydrogels are complex mixtures of water, ions, carbohydrates, and proteins. Uncertainty surrounding their composition and how interactions between components contribute to mucus function complicates efforts to exploit their properties. There is substantial interest in commercializing mucus from the garden snail, Cornu aspersum, for skincare, drug delivery, tissue engineering, and composite materials. C. aspersum secretes three mucus-one shielding the animal from environmental threats, one adhesive mucus from the pedal surface of the foot, and another pedal mucus that is lubricating. It remains a mystery how compositional differences account for their substantially different properties. Here, we characterize mucus proteins, glycosylation, ion content, and mechanical properties that could be used to provide insight into structure-function relationships through an integrative "mucomics" approach. We identify macromolecular components of these hydrogels, including a previously unreported protein class termed Conserved Anterior Mollusk Proteins (CAMPs). Revealing differences between C. aspersum mucus shows how considering structure at all levels can inform the design of mucus-inspired materials.


Vaccination coverage among children under two years of age based on electronic immunization registry in Southern Brazil.
To evaluate the immunization program for 12 and 24-month-old children based on electronic immunization registry. A descriptive study of a random sample of 2,637 children born in 2002 living in the city of Curitiba, Southern Brazil was performed. Data was collected from local electronic immunization registers and the National Live Birth Information System, as well as from a household survey for cases with incomplete records. Coverage at 12 and 24 months was estimated and analyzed according to the socioeconomic characteristics of each administrative district and the child's enrollment status in the health care service. The coverage, completeness, and record duplication in the registry were analyzed. Coverage of immunization was 95.3% at 12 months, with no disparities among administrative districts, and 90.3% at 24 months, with higher coverage in a district with lower socioeconomic conditions (p < 0.01). The proportion of vaccines, according to type, given before and after the recommended age reached 0.9% and 32.2%, respectively. In the surveyed sample, electronic immunization registry coverage was 98%, underreporting of vaccine doses was 11%, and record duplication was 20.6%. Groups with highest coverage included children with permanent records, children with three or more appointments through the National Unified Health Care System, and children seen within Primary Health Care Facilities fully adopting the Family Health Strategy. Vaccination coverage in Curitiba was high and homogeneous among districts, and health service enrollment status was an important factor in these results. The electronic immunization registry was a useful tool for monitoring vaccine coverage; however, it will be important to determine cost-effectiveness prior to wide-scale adoption by the National Immunization Program.


Detection of ROS1 rearrangement in non-small cell lung cancer: current and future perspectives.
ROS1 rearrangement characterizes a small subset (1%-2%) of non-small cell lung cancer and is associated with slight/never smoking patients and adenocarcinoma histology. Identification of ROS1 rearrangement is mandatory to permit targeted therapy with specific inhibitors, demonstrating a significantly better survival when compared with conventional chemotherapy. Detection of ROS1 rearrangement is based on in situ (immunohistochemistry, fluorescence in situ hybridization) and extractive non-in situ assays. While fluorescence in situ hybridization still represents the gold standard in clinical trials, this technique may fail to recognize rearrangements of ROS1 with some gene fusion partner. On the other hand, immunohistochemistry is the most cost-effective screening technique, but it seems to be characterized by low specificity. Extractive molecular assays are expensive and laborious methods, but they specifically recognize almost all ROS1 fusions using a limited amount of mRNA even from formalin-fixed, paraffin-embedded tumor tissues. This review is a discussion on the present and futuristic diagnostic scenario of ROS1 identification in lung cancer.


[Anti-inflammatory activity of 5 copper complexes].
The anti-inflammatory activity of five copper complexes is shown in the cotton wad granuloma test in Rats. The activity due to copper seems to be only modulated by the ligands in the complexes studied.


Effect of adrenaline on basal and ovine corticotrophin-releasing factor-stimulated ACTH secretion in man.
Six normal male subjects were given, in single blind random order on six separate occasions, i.v. bolus doses of synthetic ovine corticotrophin-releasing factor-41 (oCRF-41; 25 and 50 micrograms) with and without adrenaline (3 micrograms/min) i.v. for 150 min, the adrenaline infusions alone and saline placebo. The adrenaline infusions resulted in plasma adrenaline concentrations of 4.33 +/- 0.82 (S.E.M.) nmol/l and were associated with an increase in blood glucose, heart rate and systolic blood pressure and a reduction of diastolic blood pressure. Despite these evident biological effects at several sites, there was no stimulation of plasma ACTH or cortisol by adrenaline in comparison with the effect of saline, and no enhancement of the stimulatory effect of either dose of oCRF-41 on ACTH or cortisol secretion. The ACTH response to 50 micrograms oCRF-41 was greater than that to 25 micrograms, indicating that the 25 micrograms dose of oCRF-41 was submaximal and capable of further enhancement. As the plasma adrenaline concentrations during the adrenaline infusions reached the upper limit of the physiological range of plasma adrenaline in man, yet failed to enhance the ACTH or cortisol responses to a submaximal dose of oCRF-41, we conclude that circulating adrenaline neither exerts a direct stimulatory effect on pituitary corticotrophs nor enhances the effect of CRF under physiological circumstances. The adrenaline infusions attenuated the ACTH and cortisol responses to oCRF-41 and were associated with a transient reduction of basal concentrations of both hormones.


Severe and enduring anorexia nervosa: Update and observations about the current clinical reality.
Several objectives underlie the current article. First, to review historical diagnostic issues and clinical strategies for treating SE-AN. Second, to provide an overview of recent evidence informed strategies and clinical innovations for the treatment of SE-AN. Third, based on the authors' collective clinical and research experience, we offer eight observations that we believe capture the current clinical experience of patients with SE-AN. Some of these observations represent empirically testable hypotheses, but all are designed to generate a meaningful discussion about the treatment of this group of individuals with eating disorders. Finally, we hope to call clinicians, scientists, professional organizations, advocates, and policy makers to action to attend to critical issues related to the care of individuals with SE-AN. We believe that an international discussion could clarify areas of need for these patients and identify opportunities for clinical innovation that would enhance the lives of individuals with SE-AN and their families.


Can Surgery Be Avoided? Exclusive Antibiotic Treatment for Pelvic Actinomycosis.
Pelvic actinomycosis is an uncommon, slowly progressing granulomatous infection that has been associated with the presence of intrauterine devices. Due to its unspecific clinical and radiologic findings, it can mimic pelvic or intra-abdominal malignancy leading to mutilating surgery of high morbidity. Rarely, diagnosis is made preoperatively and in most cases surgical intervention is necessary. The patient in our case is a 42-year-old female with an IUD for 15 years diagnosed with pelvic actinomycosis. Patient was uniquely diagnosed preoperatively through paracentesis and treated conservatively with prolonged antibiotic therapy and without any type of surgical intervention. Follow-up at 1 year showed almost complete radiologic resolution of the inflammatory mass, nutritional recovery, and absence of symptoms. Pelvic actinomycosis can be successfully diagnosed and treated medically without surgical interventions.


Simplified fabrication of laminated paper-based analytical device (LPAD) with color-palette mobile app for analysis of salicylic acid in pharmaceutical products.
In this work, we present for the first time, a simplified fabrication of a laminated paper-based analytical device (LPAD) with a free-mobile app, Palette Cam, for image analysis. A filter paper is cut in a rectangular shape (9 × 3 cm) and placed between a top laminating sheet with punched holes and a bottom laminating sheet. The holes allow accessibility of liquid on the paper. Thermal lamination is then employed to complete the fabrication of LPAD. Our simplified design reduces a tedious alignment of small pieces of paper to the holes. We demonstrated the LPAD with an analysis of salicylic acid in pharmaceutical products. Each 4 μL of ferric reagent and sample was dispensed on the LPAD. Smartphone was used to capture images. The RGB (red green blue) color intensity from the Palette Cam was converted into a logarithm color ratio. Our LPAD is simplified, cost-effective and able to be a portable device.


Lectin-binding affinities of human epidermal tumors and related conditions.
Paraffin-embedded sections from a variety of epidermal lesions were stained with fluorescein isothiocyanate-labeled concanavalin A and examined by a fluorescence microscope. Seventy-six normal, hyperplastic, and neoplastic tissues were examined. Lectin binding was demonstrated in all malignant tumors, the fluorescence being confined to the plasma membrane of the tumor cells. Normal and hyperplastic tissues either failed to stain or showed a grossly diminished level of fluorescence. The distinction between malignant and normal of hyperplastic cells was clear-cut and definite.


Genome Editing in Clostridium saccharoperbutylacetonicum N1-4 with the CRISPR-Cas9 System.
Clostridium saccharoperbutylacetonicum N1-4 is well known as a hyper-butanol-producing strain. However, the lack of genetic engineering tools hinders further elucidation of its solvent production mechanism and development of more robust strains. In this study, we set out to develop an efficient genome engineering system for this microorganism based on the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated 9 (CRISPR-Cas9) system. First, the functionality of the CRISPR-Cas9 system previously customized for Clostridium beijerinckii was evaluated in C. saccharoperbutylacetonicum by targeting pta and buk, two essential genes for acetate and butyrate production, respectively. pta and buk single and double deletion mutants were successfully obtained based on this system. However, the genome engineering efficiency was rather low (the mutation rate is <20%). Therefore, the efficiency was further optimized by evaluating various promoters for guide RNA (gRNA) expression. With promoter P J23119 , we achieved a mutation rate of 75% for pta deletion without serial subculturing as suggested previously for C. beijerinckii Thus, this developed CRISPR-Cas9 system is highly desirable for efficient genome editing in C. saccharoperbutylacetonicum Batch fermentation results revealed that both the acid and solvent production profiles were altered due to the disruption of acid production pathways; however, neither acetate nor butyrate production was eliminated with the deletion of the corresponding gene. The butanol production, yield, and selectivity were improved in mutants, depending on the fermentation medium. In the pta buk double deletion mutant, the butanol production in P2 medium reached 19.0 g/liter, which is one of the highest levels ever reported from batch fermentations.IMPORTANCE An efficient CRISPR-Cas9 genome engineering system was developed for C. saccharoperbutylacetonicum N1-4. This paves the way for elucidating the solvent production mechanism in this hyper-butanol-producing microorganism and developing strains with desirable butanol-producing features. This tool can be easily adapted for use in closely related microorganisms. As also reported by others, here we demonstrated with solid data that the highly efficient expression of gRNA is the key factor determining the efficiency of CRISPR-Cas9 for genome editing. The protocol developed in this study can provide essential references for other researchers who work in the areas of metabolic engineering and synthetic biology. The developed mutants can be used as excellent starting strains for development of more robust ones for desirable solvent production.


Effectiveness of sodium bicarbonate combined with hydrogen peroxide and CPP-ACPF in whitening and microhardness of enamel.
This study investigated the effects of sodium bicarbonate (NaHCO3) combined with 1.5% hydrogen peroxide (H2O2) and casein phosphopeptide amorphous calcium phosphate fluoride (CPP-ACPF) on color and microhardness of enamel. Seventy-five bovine incisors were immersed in a tea solution for 7.5 days. The specimens were randomly divided into five groups according to the whitening agent applied: 1) 94% NaHCO3, 2) a blend of 94% NaHCO3 and CPP-ACPF, 3) a blend of 94% NaHCO3 and 1.5% H2O2, 4) a blend of 94% NaHCO3, 1.5% H2O2 and CPP-ACPF, 5) control. The whitening procedure was performed for 10 times over 10 days. At each day, the buccal surfaces were covered with whitening agents for 5 minutes and then brushed for 30 seconds. After the 10 days, the teeth were again immersed in a tea solution for 10 minutes. Color assessment was performed at baseline (T1), after the first staining process (T2), after the whitening procedure (T3), and after the second staining process (T4). Finally, the specimens were subjected to microhardness test. There was a statistically significant difference in the color change between T2 and T3 stages among the study groups (p<0.05), with the greatest improvement observed in group 4. Microhardness was significantly greater in groups 2 and 4, as compared to the other groups (p<0.05). The combination of 94% NaHCO3, 1.5% H2O2 and CPP-ACPF was effective in improving color and microhardness of teeth with extrinsic stains and could be recommended in the clinical situation.


Phylogenetic Reconstruction and Functional Characterization of the Ancestral Nef Protein of Primate Lentiviruses.
Nef is an accessory protein unique to the primate HIV-1, HIV-2, and SIV lentiviruses. During infection, Nef functions by interacting with multiple host proteins within infected cells to evade the immune response and enhance virion infectivity. Notably, Nef can counter immune regulators such as CD4 and MHC-I, as well as the SERINC5 restriction factor in infected cells. In this study, we generated a posterior sample of time-scaled phylogenies relating SIV and HIV Nef sequences, followed by reconstruction of ancestral sequences at the root and internal nodes of the sampled trees up to the HIV-1 Group M ancestor. Upon expression of the ancestral primate lentivirus Nef protein within CD4+ HeLa cells, flow cytometry analysis revealed that the primate lentivirus Nef ancestor robustly downregulated cell-surface SERINC5, yet only partially downregulated CD4 from the cell surface. Further analysis revealed that the Nef-mediated CD4 downregulation ability evolved gradually, while Nef-mediated SERINC5 downregulation was recovered abruptly in the HIV-1/M ancestor. Overall, this study provides a framework to reconstruct ancestral viral proteins and enable the functional characterization of these proteins to delineate how functions could have changed throughout evolutionary history.


Multifunctional optical correlator for picosecond ultraviolet laser pulse measurement.
A compact multifunctional optical correlator system for pulse width measurement of ultrashort ultraviolet (UV) pulses has been designed and experimentally demonstrated. Both autocorrelation and cross-correlation functions are measured using a single nonlinear crystal, and the switching between two measurements requires no adjustment of phase matching and detector. The system can measure UV pulse widths from sub-picoseconds to 100 ps, and it involves no auxiliary pulse in the measurement. The measurement results on a burst-mode picosecond UV laser show a high-quality performance on speed, accuracy, resolution, and dynamic range. The proposed correlator can be applied to measure any ultrashort UV pulses produced through sum-frequency generation or second-harmonic generation.


Sex differences in parent-reported executive functioning and adaptive behavior in children and young adults with autism spectrum disorder.
This study is the largest to date examining executive function and adaptive skills in females with autism spectrum disorder (ASD). Its primary aim was to utilize parent ratings of real-world executive functioning and adaptive behavior to better understand whether females with ASD differ from males with ASD in these areas of everyday functioning. We compared 79 females with ASD to 158 males with ASD (ages 7-18) who were statistically matched on age, IQ, and level of ADHD or ASD traits. All participants were assessed using the Behavior Rating Inventory of Executive Function (BRIEF) and a subset (56 females and 130 males) also received the Vineland Adaptive Behavior Scales (VABS). Females were rated by parents as having greater problems with executive function on the BRIEF. Parents also rated females as exhibiting more difficulties than males on the Daily Living Skills domain of the VABS. There was a correlation between increased global EF difficulty and decreased adaptive ability in both males and females. Our results indicate relative weaknesses for females compared to males diagnosed with ASD on executive function and daily living skills. These differences occur in the absence of sex differences in our sample in age, IQ, clinician ratings of core ASD symptomatology, parent ratings of ADHD symptoms, and parent-reported social and communication adaptive skills on the VABS. These findings indicate specific liabilities in real world EF and daily living skills for females with ASD and have important implications for targeting their treatments. Autism Res 2017, 10: 1653-1662. © 2017 International Society for Autism Research, Wiley Periodicals, Inc.


Spin-labeled alkylphospholipids in a dipalmitoylphosphatidylcholine bilayer: molecular dynamics simulations.
Molecular dynamics simulation has been performed to investigate the structural properties of perifosine and its synthetic spin-labeled alkylphospholipid analogues. The conformations adopted by these compounds in water and in a dipalmitoylphosphatidylcholine bilayer as a function of the presence and position of the N-oxyl-4',4'-dimethyloxazolidine ring (doxyl group) have been investigated by all-atom molecular dynamics. No predominant conformation was observed in water, but the molecules adopt specific orientations and conformations in the lipid bilayer. As is expected, alkyl chains tend to insert into the hydrophobic core, while charged groups stay at the lipid-water interface. A doxyl group in the middle of the alkyl chain moves up to the interface region, thus preventing adoption of the extended conformation. Compounds with a doxyl group close to the polar head group adopt conformations similar to that of unlabeled perifosine within the first nanoseconds of simulation. When the doxyl group is at the end of alkyl chain, the spin-labeled molecule needs more time to reach equilibrium. These results indicate a considerable effect of the doxyl position within the alkyl chain on its localization in the lipid bilayer and can be extended further to other similar spin probes used in the electron paramagnetic resonance spectroscopy of biological membranes.


Role of P2X7 Receptor in an Animal Model of Mania Induced by D-Amphetamine.
The objective of this study was to explore the association between the P2X7 purinergic receptor (P2X7R) and neuroinflammation using a preclinical model of acute bipolar mania. We analyzed the modulatory effects of P2X7R agonist (3'-O-(4-benzoyl)benzoyl-adenosine 5'-triphosphate, BzATP) and antagonists (brilliant blue, BBG and 3-[[5-(2,3 dichlorophenyl)-1H-tetrazol-1-yl]methyl]pyridine hydrochloride, A438079) on assessments related to behavior (locomotor activity), neuroinflammation (interleukin-1 beta, IL-1β; tumor necrosis factor alpha, TNF-α; and interleukin- 6, IL-6), oxidative stress (thiobarbituric acid reactive substances, TBARS) and neuroplasticity (brain-derived neurotrophic factor, BDNF) markers in a pharmacological model of mania induced by acute and chronic treatment with D-amphetamine (AMPH) (2 mg/kg) in mice. An apparent lack of responsiveness to AMPH was observed in terms of the locomotor activity in animals with blocked P2X7R or with genetic deletion of P2X7R in knockout (P2X7R(-/-)) mice. Likewise, P2X7R participated in the AMPH-induced increase of the proinflammatory and excitotoxic environment, as demonstrated by the reversal of IL-1β, TNF-α, and TBARS levels caused by P2X7R blocking. Our results support the hypothesis that P2X7R plays a role in the neuroinflammation induced by AMPH in a preclinical model of mania, which could explain the altered behavior. The present data suggest that P2X7R may be a therapeutic target related to the neuroinflammation reported in bipolar disorder.


The effect of an alternative reduced-dose infant schedule and a second year catch-up schedule with 7-valent pneumococcal conjugate vaccine on pneumococcal carriage: a randomized controlled trial.
The 7-valent pneumococcal conjugate vaccine (PCV7) was initially licensed for use as 3 infant doses and a booster (3+1). However, 2 infant doses plus a booster schedules only (2+1) are widely used. We compared the effect of these two schedules on pneumococcal carriage in young children. We also assessed the effect of a 2-dose schedule in the second year ("catch-up" schedule; 0+2). Subjects (n=733) were randomized to the 2+1 (4, 6, 12 m), 3+1 (2, 4, 6, 12 m) or 0+2 (12, 18 m) schedules. Blood samples for serotype-specific IgG (SSIgG) determination were obtained at 2, 7, 13, 19 months, and nasopharyngeal+oropharyngeal pneumococcal cultures were obtained at 2, 4, 6, 7, 12, 13, 18, 19, 24, 30 months. After primary infant PCV7 series, SSIgG was significantly lower for four out of seven serotypes in children receiving 2 doses compared to 3 doses, particularly for serotypes 6B and 23F. This was associated with a higher acquisition and prevalence rates of vaccine serotype carriage in the 2-dose group, particularly serotypes 6A and 6B. After the booster dose at 12 months of age, most differences were not significant anymore. A single PCV7 dose at age 12 months in previously unvaccinated subjects ("catch up" schedule) resulted in poor SSIgG concentrations for three out of seven serotypes, resulting in higher acquisition and prevalence rates of vaccine serotypes (grouped) compared to infants receiving a booster dose at 12 months (2+1 and 3+1 groups). Similarly, serotypes 6B and 6A also showed significantly higher carriage rates after a single dose at 12 months. After the second catch-up dose at 18 months, the rates were similar to those in the 2+1 or 3+1 schedules, except for serotype 6A. Three infant doses seem to better protect against PCV7-serotype acquisition and carriage than two. However, after booster, most of these differences disappear. A 2-dose second year catch-up campaign may enhance the reduction of PCV7-serotype spread in the community.


Conceptual and design essentials for evaluating mechanisms of change.
Considerable progress has been made toward the development of evidence-based treatments for a wide range of psychological disorders; however, little is known about the mechanisms through which these treatments actually lead to clinical change. Although the use of traditional randomized controlled treatment designs and tests of statistical mediation have significantly advanced understanding of psychological treatments, they are insufficient to test mechanisms of change. This article outlines the conceptual and methodological requirements for evaluating mechanisms of change, highlights the importance of such a focus, and offers specific recommendations for research aimed at elucidating change mechanisms. Conceptualizing and conducting studies that test mechanisms of change requires substantial modifications to traditional research designs, but doing so will significantly enhance scientific understanding as well as the efficiency and effectiveness of clinical interventions.


Vitamin C Promotes Apoptosis and Cell Cycle Arrest in Oral Squamous Cell Carcinoma.
Oral squamous cell carcinoma (OSCC) is currently ranked as the eighth most prevalent type of cancer. Despite recent advances in cancer research, the 8-year survival rate for oral squamous cell carcinoma remains only 50-60%. Therefore, markers for early detection, identification of efficient chemotherapeutic agents, and post-therapeutic monitoring are the immediate needs. With this background, this study was designed to investigate the anticancer effects of vitamin C (VC) in oral squamous cell carcinoma. Our results showed that VC had an anticancer effect on the oral squamous cell lines used in this study. VC also showed an inhibitory effect on xenograft tumors in nude mice in vitro and had a synergistic effect with cisplatin to induce cell apoptosis. Mechanistically, VC caused a significant increase in the levels of reactive oxygen species (ROS), which led to induced genotoxic (DNA damage) and metabolic (ATP depletion) stresses, inhibited Bcl-2 expression, and promoted Bax expression and caspase-3 cleavage. VC also caused cell cycle arrest at the G0/G1 phase in OSCC cells, which is related to the activation of tumor suppressor p53 and cyclin-dependent kinase inhibitor p21. In conclusion, VC bears considerable therapeutic potential for the treatment of oral squamous cell carcinoma.


Human organoids are superior to cell culture models for intestinal barrier research.
Loss of intestinal epithelial barrier function is a hallmark in digestive tract inflammation. The detailed mechanisms remain unclear due to the lack of suitable cell-based models in barrier research. Here we performed a detailed functional characterization of human intestinal organoid cultures under different conditions with the aim to suggest an optimized ex-vivo model to further analyse inflammation-induced intestinal epithelial barrier dysfunction. Differentiated Caco2 cells as a traditional model for intestinal epithelial barrier research displayed mature barrier functions which were reduced after challenge with cytomix (TNFα, IFN-γ, IL-1ß) to mimic inflammatory conditions. Human intestinal organoids grown in culture medium were highly proliferative, displayed high levels of LGR5 with overall low rates of intercellular adhesion and immature barrier function resembling conditions usually found in intestinal crypts. WNT-depletion resulted in the differentiation of intestinal organoids with reduced LGR5 levels and upregulation of markers representing the presence of all cell types present along the crypt-villus axis. This was paralleled by barrier maturation with junctional proteins regularly distributed at the cell borders. Application of cytomix in immature human intestinal organoid cultures resulted in reduced barrier function that was accompanied with cell fragmentation, cell death and overall loss of junctional proteins, demonstrating a high susceptibility of the organoid culture to inflammatory stimuli. In differentiated organoid cultures, cytomix induced a hierarchical sequence of changes beginning with loss of cell adhesion, redistribution of junctional proteins from the cell border, protein degradation which was accompanied by loss of epithelial barrier function. Cell viability was observed to decrease with time but was preserved when initial barrier changes were evident. In summary, differentiated intestinal organoid cultures represent an optimized human ex-vivo model which allows a comprehensive reflection to the situation observed in patients with intestinal inflammation. Our data suggest a hierarchical sequence of inflammation-induced intestinal barrier dysfunction starting with loss of intercellular adhesion, followed by redistribution and loss of junctional proteins resulting in reduced barrier function with consecutive epithelial death.


[Long-term results of surgical treatment of the patients with Leriche syndrome].
The paper analyzed results of surgical treatment of the 94 patients with Leriche syndrome and researched the long-term results of 52 (55.3%) cases. Following 5 years patency after aortofemoral reconstructions was observed in 34 (65.4%) cases. Lower extremity preservation was observed in 35 (67.3%) patients. Five year survival was 76.9%, and only 53.8% patients received adequate postoperative diagnostic monitory and therapy.


NMR structural study of the prototropic equilibrium in solution of Schiff bases as model compounds.
An NMR titration method has been used to simultaneously measure the acid dissociation constant (pKa) and the intramolecular NHO prototropic constant ΔKNHO on a set of Schiff bases. The model compounds were synthesized from benzylamine and substituted ortho-hydroxyaldehydes, appropriately substituted with electron-donating and electron-withdrawing groups to modulate the acidity of the intramolecular NHO hydrogen bond. The structure in solution was established by 1H-, 13C- and 15N-NMR spectroscopy. The physicochemical parameters of the intramolecular NHO hydrogen bond (pKa, ΔKNHO and ΔΔG°) were obtained from 1H-NMR titration data and pH measurements. The Henderson-Hasselbalch data analysis indicated that the systems are weakly acidic, and the predominant NHO equilibrium was established using Polster-Lachmann δ-diagram analysis and Perrin model data linearization.


Function-based gene identification using enzymatically generated normalized shRNA library and massive parallel sequencing.
As a general strategy for function-based gene identification, an shRNA library containing approximately 150 shRNAs per gene was enzymatically generated from normalized (reduced-redundance) human cDNA. The library was constructed in an inducible lentiviral vector, enabling propagation of growth-inhibiting shRNAs and controlled activity measurements. RNAi activities were measured for 101 shRNA clones representing 100 human genes and for 201 shRNAs derived from a firefly luciferase gene. Structure-activity analysis of these two datasets yielded a set of structural criteria for shRNA efficacy, increasing the frequencies of active shRNAs up to 5-fold relative to random sampling. The same library was used to select shRNAs that inhibit breast carcinoma cell growth by targeting potential oncogenes. Genes targeted by the selected shRNAs were enriched for 10 pathways, 9 of which have been previously associated with various cancers, cell cycle progression, or apoptosis. One hundred nineteen genes, enriched through this selection and represented by two to six shRNAs each, were identified as potential cancer drug targets. Short interfering RNAs against 19 of 22 tested genes in this group inhibited cell growth, validating the efficiency of this strategy for high-throughput target gene identification.